Gene Literature Dashboard

Nanoparticles in physiological environments acquire a biomolecular corona that defines their biological identity, mediating immune system recognition and accelerating blood clearance of the nanoparticles. Typically, low-fouling materials are chosen to minimize protein adsorption and thereby immune system responses, contributing to stealth in blood. However, absolute prevention of the biomolecular corona remains tantalizingly out of reach. Herein, it is proposed to leverage the biomolecular corona rather than preventing its formation, in order to overcome immune responses toward nanoparticles. Low-fouling and stealthy poly(ethylene glycol)(PEG) nanoparticles are used, with a functional biomolecular corona enriched with anticoagulant heparin-antithrombin III (HEP-ATIII) complexes that can mitigate undesirable immune responses. Through immune response evaluations and proteomic analyses are used to ascertain the low-fouling, stealthy character of the nanoparticles, similar to that of PEG nanoparticles. However, PEG nanoparticles alone induce coagulation responses in human blood, which are mitigated by pre-enrichment of the biomolecular corona with HEP-ATIII complexes. This shows that coagulation is another factor to be considered in the design of materials for nanomedicine and that the low-fouling and stealthy properties do not directly translate to hemocompatibility. These findings highlight the potential of biomolecular corona engineering to address key challenges in the field, toward developing safer, efficacious therapeutic nanomaterials.

Despite significant recent progress in the field of mass spectrometry (MS)-based top-down proteomics (TDP), the analysis of limited samples is still a major challenge. Here, we explored the potential of ultralow flow (ULF) liquid chromatography (LC) porous layer open tubular (PLOT) columns interfaced with MS via high-field asymmetric waveform ion mobility spectrometry (FAIMS) to enable high-sensitivity TDP analysis of small populations of mammalian cells. The developed robust and easy-to-use platform delivered high reproducibility of retention times (RSD < 0.4%) and high separation performance for intact proteins (∼14-s peak full width at half-maximum and peak capacity of >125 for a 60 min effective gradient). The FAIMS-based experiments resulted in a ∼2-fold increase in identifications compared to the control experiments for ∼200 HeLa cell aliquots, i.e., 819 vs 454 proteins and 2645 vs 1305 proteoforms, respectively. The pilot ULF LC-MS analysis of six HeLa cells yielded 29 ± 3 proteins and 38 ± 2 proteoforms, on average, and a total of 44 proteins and 68 proteoforms. Data revealed a high degree of acetylation, methylation, phosphorylation, glycosylation, lactylation, and other relevant post-translational modifications. Notably, the presented protein identification results for limited samples are comparable to those of recent large-scale TDP studies of bulk samples, demonstrating the potential to enable informative single-cell TDP profiling.

<h4>Background</h4>Screening for BCR-ABL1 kinase domain (KD) mutations is routinely implemented in cases of treatment failure for chronic myeloid leukaemia and Philadelphia-positive acute lymphoblastic leukaemia. However, timely deciphering their clonal relationship via mutation profiling that requires identification of mutation types, quantification of mutant abundance, and differentiation between compound and polyclonal mutations (CMs and PMs), remains difficult during therapy.<h4>Methods</h4>Herein, we established a protocol that identified mutation types and further distinguished clonal relationships by combining mini-sequencing of MeltArray with allele segregation of droplet digital PCR (ddPCR).<h4>Results</h4>The analysis showed that 78 samples (18.93%) were mutant, of which 50 (64.1%) harboured single mutations, and 28 (35.9%) contained multiple mutations, including double-, triple-, quadruple- and hepta-mutants. These results agreed with NGS, except one sample with F317L and L324Q mutations, where L324Q was beyond MeltArray's scope. Among cases containing multiple mutations, 85.71% were PMs, 10.71% were CMs, and 3.57% were mixed CMs and PMs. Retrospective analysis revealed that clonal relationships in BCR-ABL1 KD mutations were highly dynamic during therapy.<h4>Conclusions</h4>The MeltArray-ddPCR protocol enables dynamic profiling of BCR-ABL1 KD mutations to determine clonal status, improving prediction of drug susceptibility and leukaemia outcomes. In this study, we performed a retrospective analysis of 539 samples from 365 leukaemia patients. We developed a comprehensive BCR-ABL1 kinase domain mutation screening protocol that includes pre-amplification, mutation screening, and differentiation between compound mutations (CMs) and polyclonal mutations (PMs). By integrating MeltArray and ddPCR technologies, this protocol enables dynamic monitoring of changes in mutation levels and clonal evolution. It also offers companion diagnostics based on mutational profiles to guide precision therapy for leukaemia patients.

<h4>Background & aims</h4>Depression and osteoporosis are common among middle-aged and older adults, both impacting morbidity and quality of life. Their shared risk factors suggest a potential link, but this relationship remains underexplored. This study aimed to assess the association between depressive symptoms and osteoporosis in two large cohorts: the National Health and Nutrition Examination Survey (NHANES, 2005-2010) and Health and Retirement Study (HRS, 2012).<h4>Methods</h4>We analyzed data from adults aged ≥50 years in NHANES (n = 3612) and HRS (n = 4307). Depressive symptoms were measured using PHQ-9 in NHANES and CES-D 8 in HRS, while osteoporosis was defined by self-reported diagnosis and medication use. Generalized linear models were used to evaluate the association, adjusting for demographic, lifestyle, and comorbidity factors. Subgroup analyses and sensitivity tests were conducted to explore effect modifiers and result robustness.<h4>Results</h4>Depressive symptoms were positively associated with osteoporosis in both cohorts (NHANES: adjusted OR = 1.061, 95% CI: 1.034-1.088; HRS: adjusted OR = 1.063, 95% CI: 1.014-1.115). Significant associations were observed across subgroups, with stronger effects in individuals with diabetes and arthritis. Sex differences showed higher odds ratios for men in HRS. The relationship exhibited a linear trend, with increasing risk as depressive severity intensified.<h4>Conclusion</h4>Depressive symptoms (PHQ-9/CES-D8) significantly associate with higher osteoporosis risk in older adults, with consistent cross-sectional findings across populations. While causality remains unclear, results support clinical bone density monitoring in depression care and depression screening in osteoporosis management. Future longitudinal studies should clarify mechanisms, while public health strategies should integrate mental-bone health prevention approaches.

Huntington's disease (HD) is caused by the expansion of poly-glutamine repeats in the Huntingtin (<i>Htt</i>) gene and is associated with a wide variety of motor and physiological (sleep, metabolism, etc.) perturbations. Studies from diverse model organisms have proposed that modulation of autophagy (a key protein homeostatic pathway) can mitigate the toxic effects of mutant HTT protein. However, consistent changes are not observed across studies, and the improvements in phenotypes can be associated with changes in specific circuits/neurons affected by the mutant HTT protein. They suggest that not all neurons respond effectively to autophagy modulation. Hence, it remains to be understood whether diverse circuits/neurons affected by mutant HTT protein respond effectively to this intervention. Using a genetic approach, we expressed mutant HTT protein independently in diverse sets of neurons in male <i>Drosophila melanogaster</i> and asked whether genetic modulation of autophagy pathway through <i>Atg8a</i> overexpression can mitigate the toxic effect of mutant HTT protein. We found that in male flies, not all neurons/circuits expressing mutant HTT protein respond effectively to ATG8a protein. Circadian neurons and neurons regulating carbohydrate and lipid metabolism (<i>Dilp2</i> ^+ve) showed improvement, while motor and neurons responding to temperature changes showed no improvement. Using cellular markers we also showed that these phenotypes can be attributed to specific changes in mutant HTT and Ref(2)P proteins (autophagy marker). Our study suggests that not all circuits respond effectively to autophagy modulation and suggests a potential cause for low success of autophagy modulators in clinical trials..

<b>Background</b>: Zinc finger proteins (ZNFs), functioning as pervasive transcriptional modulators, serve as pivotal mediators of tumorigenesis and malignant advancement. However, the mechanistic contributions of these epigenetic orchestrators to lung adenocarcinoma pathogenesis remain incompletely characterized. <b>Methods</b>: To elucidate zinc finger proteins' biological significance in lung adenocarcinoma (LUAD) pathogenesis, we first extracted relevant transcriptional data from TCGA. After preliminary screening with univariate Cox regression, a LASSO algorithm was applied to optimize the risk score model, incorporating key zinc finger protein markers. For independent validation, we accessed GEO dataset GSE68465, applying identical analytical protocols to confirm model generalizability. We performed multivariable Cox regression to identify independent predictors of clinical outcomes after adjusting for confounding variables. Cell-based validation included (1) comparative analysis of zinc finger protein expression across LUAD/normal cell models and (2) technical verification using standardized qRT-PCR protocols. <b>Results</b>: Following rigorous bioinformatics screening comprising differential expression and survival analysis, the final 21-zinc finger protein cohort was selected for risk score algorithm development aimed at clinical outcome prediction. Stratification based on computed risk scores revealed markedly superior survival outcomes in the low-risk cohort compared to high-risk patients. Comparative analysis revealed overall concordance in the transcriptional profiles of eight ZNFs (|coef| > 0.1) across experimental cell systems and TCGA datasets. <b>Conclusions</b>: Collectively, the prognostic framework incorporating zinc finger proteins demonstrates biomarker utility in lung adenocarcinoma survival prediction, while offering novel avenues for molecular target discovery in therapeutic strategies against this malignancy.

Hepatobiliary diseases, which include disorders of the liver, gallbladder, and bile ducts, remain a major global health concern. A significant proportion of deaths worldwide are attributed to hepatic diseases, accounting for 4% of the total global mortality in 2023. Among benign hepatobiliary diseases, metabolic dysfunction-associated steatotic liver disease is the most prevalent liver pathology, with a concerning rise in incidence, while it is recognized as the leading cause of liver transplantation in the United States. However, there is a notable rise over time in cases of autoimmune hepatobiliary disorders, including autoimmune hepatitis, primary biliary cholangitis, and primary sclerosing cholangitis. Meanwhile, hepatocellular carcinoma still remains the most frequently diagnosed hepatobiliary malignancy, constituting the third leading cause of malignancy-related mortality globally. Meanwhile, cholangiocarcinoma and gallbladder cancer are the second and third most common hepatobiliary malignancies, respectively, both exhibiting highly aggressive malignant behavior. Despite the notable advances in biomarkers and the development of therapeutic tools, early diagnosis and monitoring are considered pivotal for the management of the aforementioned pathologies. The development of new non-invasive biomarkers that can effectively identify, monitor these pathologies, and guide their management is considered a necessity. Extracellular vesicles (EVs) constitute nanoparticles with several embedded cargoes, with a significant role in intercellular communication, which are considered promising biomarkers in several diseases, including viral, metabolic, autoimmune, and malignant diseases. In this review, we will shed light on the role of EVs as novel frontiers in hepatobiliary diseases.

The establishment of functional neuronal circuits critically relies on the ability of developing neurons to accurately sense and integrate a variety of guidance signals from their surrounding environment. Such signals are indeed crucial during key steps of neuronal circuit wiring, including neuronal migration and axon guidance, to guide developing neurons or extending axons towards their target destination in the developing brain. The growth cone, located at the tip of developing neurons, is a key subcellular structure in this process, that concentrates many different guidance receptors and signalling molecules and specialises in the probing and integration of extracellular signals into various guidance behaviours. Interestingly, the small primary cilium, long considered as a vestigial organelle, has progressively emerged as a cellular antenna specialised in cell signalling, and has been reported, just like the growth cone, to harbour a variety of guidance receptors. How primary cilium-elicited signals are then transduced into specific cellular processes to guide developing neurons and axons remains however obscure. In this review, we will summarise our emerging understanding of the role of primary cilium-elicited signalling pathways on neuronal guidance processes, by focusing on neuronal migration and axon guidance. We will highlight the primary cilium molecular diversity, and how it shapes the primary cilium functional versatility, allowing the ciliary compartment to instruct various guidance behaviours through the regulation of different cellular processes. We will moreover discuss current and future avenues of research, to unravel the different molecular effectors activated downstream of specific ciliary signals, and clues to be gained from studies performed in non-neuronal cells. Rising challenges of the field will also be addressed, such as the technical challenge induced by the dual subcellular localisation (<i>i.e</i>., ciliary and extra-ciliary) of many ciliary guidance receptors, and the importance of the development of new genetic/chemo-genetic/optogenetic tools. Finally, we will highlight the insight such studies will bring for our understanding of the aetiology of different disorders, including ciliopathies, neurodevelopmental and neurodegenerative disorders, but also cancer cell migration/invasion, which are associated with defective primary cilium formation and function.

Diabetic cardiomyopathy (DCM) is a complication of diabetes and is the main cause of death in diabetic patients. The regulatory networks and key players involved in the pathogenesis of diabetic cardiomyopathy are not clearly known. We selected the miRNA, protein, and metabolite fingerprints that play a significant role in DCM and manually constructed miRNA-protein-metabolite interaction networks from the miRNA-protein and protein-metabolite interaction networks. Furthermore, protein-protein, metabolite-metabolite, and protein-metabolite interaction networks were also constructed. The miRNA-protein interaction included evidence from TarBase and microarrays/HITS-CLIP. The protein-protein, metabolite-metabolite, and protein-metabolite interaction networks were obtained at high confidence scores (≥0.7 or 70%). We proposed that the miRNA-protein-metabolite interaction networks along with their intra- and inter-connected protein-protein, metabolite-metabolite, and protein-metabolite interaction networks formed by miRNA, protein, and metabolite fingerprints such as hsa-mir-122-5p, hsa-mir-30c-5p, hsa-mir-30d-5p, hsa-mir-22-3p, IL6, GSTM2, GPX3, ACADM, GSTM3, LEP, ADIPOQ, INS, CASP1, NLRP3, HADH, ACAT1, PRDX2, PRDX1, TNF, ELAVL1, SERPINA1, A2M, IGFBP7, PRDX6, APOA1, APCS, NPPA, ADAM9, GDF15, ACADVL, ECH1, FGL1, bilirubin, butyric acid (butyrate), octanoylcarnitine (octanoylcarnit.), isoleucine, leucine, alanine, glutamine, L-valine, cytidine triphosphate (ara-CTP), 7-keto-8-aminopelargonic acid (7-keto-8-amino.), creatinine, decanoylcarnitine (decanoylcarnit.), and hexanoylcarnitine (hexanoylcarnit.) are the key players and regulatory networks involved in the pathogenesis of DCM. Notably, we also proposed that the interaction networks formed by miRNA, protein, and metabolite fingerprints involved in the early stage of DCM, such as hsa-mir-122-5p, IL6, FGL1, LEP, ADIPOQ, INS, TNF, IGFBP7, GDF15, GPX3, NPPA, bilirubin, butyric acid (butyrate), and creatinine, are the potential biomarkers and therapeutic targets for the early stage of DCM. To the best of our knowledge, this is the first study of the construction of miRNA-protein-metabolite interactomes in DCM, providing insights into the pathogenesis of DCM.

Neurodegenerative diseases represent a major global cause of mortality and disability. These disorders are characterized by complex pathogenesis and currently lack effective therapeutic strategies. Iron, a vital trace element for normal brain function, has been implicated in the pathogenesis of neurodegenerative diseases via the ferroptosis pathway. Emerging evidence indicates that exercise can suppress ferroptosis directly or indirectly by regulating iron metabolism, oxidative stress, and exerkine expression, thereby conferring neuroprotection. This review summarizes current insights into the role of ferroptosis in neurodegenerative diseases and explores the mechanisms by which exercise modulates the ferroptosis pathway, offering a scientific rationale for exercise-based interventions in brain health.

Directed evolution leverages the principles of natural selection to engineer biomolecules with desired properties. Microbead-based approaches within water-in-oil emulsions have proven invaluable for high-throughput <i>in vitro</i> selections. However, highly aggregation-prone microbeads present significant challenges, including clustering, inconsistent distribution, and droplet instability. Here, we introduce a simple and cost-effective method for generating polydisperse emulsions with restored Poissonian distributions of highly aggregation-prone microbeads. This approach utilizes modified gel loader pipette tips, drawn out to create nozzles capable of disrupting microbead clusters during emulsification. Two widely utilized oil-surfactant formulations-mineral oil with Abil EM 90 and FluoSurf in HFE 7500 - were evaluated for emulsion preparation. Emulsions prepared using the modified nozzles exhibited exceptional stability, maintaining integrity during week-long incubations at 37°C, and reliably distributed microbeads into droplets in accordance with a Poissonian distribution despite the microbeads' highly aggregation-prone property.

Subarachnoid hemorrhage (SAH) is a frequently encountered critical emergency characterized by the rupturing of an unhealthy blood vessel, resulting in high mortality and disability rates. Alterations in the neurovascular unit (NVU) are closely related to the pathogenesis of SAH. Microglia, the primary innate immune cells in the brain, and astrocytes, the most abundant cells in the brain, both play crucial roles in the response to SAH-associated cerebral injuries. Recently, the crosstalk between these two cells in the pathology and treatment of central nervous system (CNS) diseases, including SAH, has been revealed. Following acute brain insult, activated microglia and astrocytes can further activate each other, contributing to amplified neuroinflammatory reactions and thus inducing secondary brain injury. This review addresses the pathophysiological mechanisms of microglia and astrocytes in SAH, including neuroinflammation, neuronal damage, blood-brain barrier (BBB) disruption, vasospasm, and hematoma clearance. In addition, the newly identified therapeutic strategies against SAH by regulating astrocytes-microglia crosstalk through targeting damage-associated molecular patterns (DAMPs), immune mediators, and their receptors are also discussed. A thorough comprehension of microglia-astrocyte communication could provide novel ideas for future research and treatment of SAH.

Since the early 20^th century, there has been extensive discussion on the intricate relationship between pathogenic infection and tumors. However, most studies on host-pathogen interactions are performed based on the <i>in-vitro</i> culture, immortalized cell lines or animal experiments. A significant challenge lies in accurately establishing a coculture model between tumors and pathogens under the three-dimensional (3D) context. Recently, the hybrid model system that incorporates 3D tumor organoids and two-dimensional cell lines have been gradually used to analyze the intricate relationship between pathogens and tumors, and several coculture techniques for tumor organoids and pathogens have also been developed. Therefore, this study systematically reviewed the preparation and identification of tumor organoids, coculture techniques with pathogens, and their clinical applications, aiming to further understand and simulate the interaction mechanism between the hosts and pathogens.

<h4>Background and aims</h4>With the world's aging population, twin epidemics of type-2 diabetes (T2D) and dementia take a great toll on the healthcare burden. T2D carries a 2-3 times greater risk of developing cognitive impairment than controls. Early identification of cognitive impairment is important as it impairs diabetes self-management, making patients prone to complications. However, data about the assessment of cognitive impairment in T2D using a comprehensive cognitive battery is sparse in India. This study was undertaken to estimate the prevalence and pattern of cognitive impairment among young and middle-aged patients with T2D.<h4>Materials and methods</h4>A cross-sectional observational study was conducted in a tertiary care teaching hospital in Kolkata (2022-2024) among 125 Bengali-speaking T2D patients with formal education > class IV, aged between 20 and 60 years. The cognitive evaluation was done using the clinical dementia rating scale, mini-mental status examination (MMSE), Montreal cognitive assessment (MoCA), and Addenbrooke's cognitive examination (ACE)-III. Statistical analyses were done by Jeffrey's Amazing Statistics Program version 0.19 with appropriate tests (Chi-squared test, Mann-Whitney U test, Spearman correlation statistics, and logistic regression). <i>P</i> value < 0.05 was considered significant.<h4>Results</h4>T2D patients reported a more subjective sensation of forgetfulness compared to the control group (<i>P</i> = 0.001). MMSE was an insufficient screening tool to distinguish between these two groups. On MoCA and ACE-III, there was a significant difference in total scores between case and control groups (MoCA, <i>P</i> = 0.012 and ACE-III, <i>P</i> < 0.001). Based on ACE-III, 59.20% of T2D patients had cognitive impairment (<i>P</i> < 0.001). The odds of having cognitive impairment in T2D were 3.72 times higher than in the control group (<i>P</i> < 0.001). There was significant impairment of memory (<i>P</i> < 0.001), fluency (<i>P</i> = 0.020), and visuospatial ability (<i>P</i> = 0.032). Females (<i>P</i> = 0.010), less education (<i>P</i> < 0.001), lower socioeconomic status (<i>P</i> < 0.001), BMI < 23 kg/m² (<i>P</i> = 0.049), peripheral neuropathy (<i>P</i> = 0.001), hypothyroidism (<i>P</i> = 0.007), anxiety (<i>P</i> < 0.001), and depression (<i>P</i> < 0.001) were significantly associated with cognitive impairment in diabetes.<h4>Conclusion</h4>This is the first study from Eastern India to use a comprehensive cognitive scale validated in the local vernacular. Cognitive impairment is prevalent among a significant portion of middle-aged, educated individuals with T2D. Cognitive evaluation should be incorporated into diabetes management from the onset, with a focus on addressing modifiable factors.

<h4>Introduction</h4>Bleeding from esophageal varices (EVs) causes a significant risk in cirrhotic patients, often leading to life-threatening complications. While screening endoscopy is the recommended method to identify those at risk, its invasive nature and limitations prompt the search for alternative predictors. Thrombocytopenia has emerged as a potential marker for assessing the presence and severity of EVs. This study aims to establish a relationship between platelet count and predicted grading of EVs in individuals with liver cirrhosis.<h4>Aim</h4>To study platelet count as a predictor of EVs in liver cirrhosis. Materials and Methods: This cross-sectional research was undertaken at KGMU, Lucknow. It encompassed newly identified cases of cirrhosis with varices of any degree detected through endoscopy. The endoscopic observations were standardized utilizing the Paquet grading system. Patients were categorized into four subgroups based on platelet count, and correlations between platelet count groups and EVs grading were examined using Spearman rank correlations. The association between platelet count and EVs grade was assessed using the Chi-square test.<h4>Results</h4>The study comprised 50 patients, with 84% (n = 42) being male, and the mean age was 45.66 ± 10.26 years. Platelet counts were categorized as follows: <50,000/uL in 36% of patients, 50,000-99,000/uL in 28%, 100,000-150,000/uL in 14%, and >150,000/uL in 22% of patients. EVs were classified as grade I in 26% of patients, grade II in 20%, grade III in 34%, and grade IV in 20% of patients. When the platelet count is <50,000/uL, the patients present grade IV varices. With a platelet count of 50,000-99,000/μL, they present grade III varices; 100,000-150,000/μL corresponds to grade II; and > 150,000/μL to grade I EVs. The mean platelet count was 223245.53/mm³ in patients with grade I varices, decreasing to 109505.42/mm³, 57345.46/mm³, and 20502.00/mm³ in patients with grade II, III, and IV varices, respectively (<i>p</i> = <0.0001). A notable negative correlation was observed between platelet count and EVs grades (<i>P</i> < 0.001).<h4>Conclusion</h4>Platelet count serves as a predictive factor for EVs grade in cirrhotic patients, with a significant negative correlation between platelet count and varices grades.

Biomolecular condensates are increasingly recognized as central regulators of numerous cellular processes. The bulk rheology of condensates (e.g., viscoelasticity) balances molecular mobility with structural stability, while the interfacial properties of condensates (e.g., interfacial tension) regulate condensate growth and their interactions with other cellular structures. Here, we review the functional roles of condensate rheology and interfacial properties, as well as diseases associated with their dysregulation. By summarizing emerging methodologies and quantitative measurements of condensate viscoelasticity and interfacial tension in the literature, we highlight key regulators of condensate material properties and discuss their implications in biology.

Epstein-Barr virus (EBV) infection is a common viral illness typically presenting with symptoms such as fever, sore throat, and lymphadenopathy. Hepatic involvement in EBV infection is usually mild and transient. However, severe cholestatic liver disease due to acute EBV hepatitis is rare, especially in young adults. Secondly, hemophagocytic lymphohistiocytosis (HLH), the abnormal activity of lymphocyte function leading to hemophagocytosis and multi-organ failure, is a rare complication of EBV. In the context of Los Angeles County, the locale of this study, the incidence of secondary nonfamilial HLH among patients over the age of 15 is reported at 0.9 cases per million annually, with epidemiological data specific to EBV-associated HLH even more notably limited. We report a case of a 20-year-old female patient presenting with fever and chills, ultimately diagnosed with acute EBV hepatitis causing severe cholestatic liver injury, with concurrent positive antimitochondrial antibody and HLH, without multi-organ failure, who showed significant improvement with the administration of N-acetylcysteine (NAC), highlighting its potential therapeutic role in EBV-associated liver diseases.

Familial adenomatous polyposis (FAP) is an autosomal dominant genetic disorder primarily caused by pathogenic mutations in the adenomatous polyposis coli (<i>APC</i>) gene. Some FAP cases are clinically diagnosed even in the absence of a family history. Both the NCCN Clinical Practice Guidelines (Version 3.2024) and the Japanese Society for Cancer of the Colon and Rectum Guidelines for the Clinical Practice of Hereditary Colorectal Cancer Guidelines (2020) recommend genetic testing for FAP cases without a family history; however, its implementation is limited due to ethical and economic considerations. Herein, we report two cases in which genetic testing was performed on patients clinically diagnosed with FAP despite the absence of a family history. Case 1: A 44-year-old woman presented with transverse colon cancer and polyposis, identified using colonoscopy. Despite having no family history of FAP, she was diagnosed with attenuated FAP (AFAP) based on the preoperative findings. The patient underwent laparoscopic total colectomy and ileorectal anastomosis, followed by adjuvant chemotherapy and surgical treatment for the pulmonary metastasis. Genetic panel testing revealed no <i>APC</i> mutation but identified a <i>SMAD9</i> mutation classified as a variant of uncertain significance. Over a follow-up period exceeding 9 years, the patient showed no recurrence of colorectal cancer or extracolonic manifestations of FAP. Case 2: A 44-year-old woman who had undergone colonoscopy since being diagnosed with polyps at the age of 29 years presented with sigmoid colon cancer and polyposis. Despite having no family history of FAP, she was diagnosed with AFAP based on the preoperative findings. The patient underwent laparoscopic total colectomy with ileostomy, followed by ileostomy closure 6 months later. Genetic testing performed the same year revealed an <i>APC</i> mutation. A CT scan at 1 year and 7 months postoperatively revealed a soft tissue mass suspected to be a desmoid tumor, and the patient is currently being followed up in the outpatient clinic. These cases emphasize the importance of genetic testing in the clinical management of FAP to ensure an accurate diagnosis and differentiation from related syndromes. Although <i>APC</i> mutations are detected in only 20-40% of patients undergoing genetic testing for FAP, <i>APC</i> mutation-negative cases are reported to have a milder phenotype. However, its genetic characteristics remain unclear. The role of <i>SMAD9</i> mutations is not yet fully understood, but identifying such mutations may deepen our understanding of genetic associations in colorectal polyposis syndromes.

Acute myeloid leukemia (AML) is a hematologic malignancy marked by uncontrolled proliferation of abnormal myeloid precursor cells, while cirrhosis involves progressive hepatic fibrosis and architectural distortion. Despite differing pathophysiologies, both can present with overlapping clinical features, complicating the diagnostic process. We present the case of a 78-year-old female with a one-month history of bilateral lower extremity edema, dyspnea, fatigue, anorexia, low-grade fever, and progressive abdominal distension. Upon admission, a comprehensive metabolic panel revealed mild transaminitis, hyperbilirubinemia, and hypoalbuminemia. Complete blood count with differential revealed moderate leukocytosis, macrocytic anemia below the transfusion threshold, severe thrombocytopenia, and 10% circulating blasts. A contrast-enhanced computed tomography scan of the abdomen and pelvis showed cirrhotic liver morphology, portal hypertension, esophageal varices, splenomegaly with infarct, and minimal ascites. Chronic liver disease workup also indicated possible hereditary hemochromatosis. Hematologic evaluation, including peripheral blood flow cytometry, confirmed AML with 11.4% blasts expressing CD34, CD117, HLA-DR, CD33, and CD13. This case highlights the rare concurrent presentation of newly diagnosed AML and cirrhosis, emphasizing the diagnostic challenges posed by overlapping symptoms and the necessity of a multidisciplinary approach for timely identification and management.

<h4>Background</h4>It is pertinent to investigate organic possibilities in acute presentation of mental illness. Iron-deficiency anemia has long been established to cause symptoms of major depressive disorder. The association between depression and hemolytic anemia and hemochromatosis is less understood.<h4>Case presentation</h4>A rare type of hemolytic anemia caused by hemoglobin (Hb) Manukau is reported in a 30-year-old female presenting with 3 weeks of severe depression, paranoia, auditory hallucinations, and suicidal ideation. The client was admitted to a psychiatric unit and received treatment with sertraline and olanzapine. Post-discharge follow up is with a community mental health team. The psychiatric history includes recurrent episodes of major depression with psychotic features, with the first occurring after a stillbirth at age 18 years. The treatment history includes electroconvulsive therapy, sertraline, and olanzapine. The medical complications of the Hb Manukau are chronic hemolytic anemia, liver disease, and hemochromatosis, requiring frequent blood transfusions and iron chelation therapy.<h4>Conclusion</h4>This rare hemoglobinopathy may contribute to the pathogenesis of recurrent depression with psychotic features, potentially through iron deposition in the brain or by other mechanisms yet to be defined.

Increasing evidence reveals that the deregulation of cellular antioxidant response with advancing age, resulting in the continuing amplification of oxidative stress-induced inflammatory response, is a pre-eminent cause for the onset of aging-related disease states, including blinding diseases. However, several safeguards, like an antioxidant defense system, are genetically in place to maintain redox homeostasis. Nonetheless, if the homeostatic capacity of such systems fails (like in aging), an inflammatory pathway elicited by excessive oxidative stress-evoked aberrant NLRP3 (NOD, LRR- and pyrin domain-containing protein 3) inflammasome activation can become pathogenic and lead to disease states. Among all known inflammasomes, NLRP3 is the most studied and acts as an intracellular sensor to detect danger(s). Upon activation, NLRP3 recruits apoptosis-associated speck-like protein containing a CARD (ASC) oligomerization and facilitates the recruitment of activated Caspase-1 (Cas-1), which results in the release of inflammatory cytokines, IL-1β and IL-18 and the activation of GasderminD, an executor of pyroptosis. NLRP3 inflammasome is tightly regulated in favor of cell health. However, when and how the activation of NLRP3 and its inflammatory components goes awry, leading to cellular derangement, and what regulatory factors are involved in the normal physiological and aging/oxidative conditions will be included in this review. Also, we address the latest findings to highlight the connection between oxidative stress, antioxidants, and NLRP3 activation as this begets aging diseases and explore the cellular pathways that are in place to regulate oxidative-induced inflammations and the pathobiological consequences of dysregulated inflammatory responses and vice versa.

<b>Introduction:</b> The interface between T cells and the tumor microenvironment, termed the 'immunological synapse', consists of multiple checkpoint protein pairs co-expressed on both sides of the synapse. mir-16, a microRNA from a widely known tumor-suppressor family of miRNAs, was previously shown by us to be downregulated in melanoma. As other miRNAs from this family have been shown to directly target checkpoint proteins, here we investigated whether miR-16 influences the expression patterns of checkpoint proteins in melanoma. <b>Methods:</b> Single-cell gene expression data from the melanoma microenvironment were retrieved from a public database. Melanoma cell lines were established from metastatic lesions and transiently transfected with an hsa-miR-16-5p-mimic RNA or a mir-16-expressing plasmid. The mRNA expression profiles were analyzed using an Affymetrix microarray. Direct targets of miR-16 were identified by luciferase reporter assays. Protein levels were assessed by Western blotting. <b>Results:</b> Bioinformatic analysis revealed that the expression levels of eight checkpoint mRNAs, known to be present on the melanoma side of the immunological synapse, were highly correlated. Four of these mRNAs contained putative binding sites for the miR-15/16 family. miR-16 expression was significantly reduced in melanoma cells, compared to normal melanocytes. Luciferase reporter assays demonstrated that miR-16 directly targets the 3' untranslated regions (3'UTRs) of CD40, CD80. The mRNAs downregulated following miR-16 overexpression were highly enriched for genes involved in autophagy, vesicle-mediated transport, and the regulation of protein membrane localization. Among these, VTI1B and SMPD1 were confirmed to be direct targets of miR-16. Transient overexpression of miR-16 resulted in a significant reduction in SMPD1 and VTI1B levels in melanoma cell lines. <b>Conclusions:</b> Our findings suggest that miR-16 potentially modulates melanoma tumorigenesis, metastasis and immunogenicity by altering the composition of checkpoint proteins at the immunological synapse and by regulating cellular pathways associated with intracellular trafficking and transmembrane protein presentation.

<b>Background:</b> Developmental delay and intellectual disability (DD/ID) are frequently accompanied by epilepsy, and growing evidence implicates variants in voltage-gated calcium channel genes in their pathogenesis. This study aimed to investigate the association of polymorphisms in <i>CACNA1A</i>, <i>CACNA1C</i>, and <i>CACNA1H</i> with DD/ID and epilepsy comorbidity in Korean children. <b>Methods:</b> We retrospectively analyzed 141 pediatric patients diagnosed with DD/ID who underwent whole-exome sequencing (WES) and were not found to have pathogenic monogenic variants. Nine single-nucleotide polymorphisms (SNPs) across <i>CACNA1A</i>, <i>CACNA1C</i>, and <i>CACNA1H</i> were selected based on functional annotation scores and prior literature. Genotype data were extracted from WES variant files, and allele and genotype frequencies were compared with control data from the gnomAD East Asian population and the Korean Reference Genome Database (KRGDB). Subgroup analyses were performed according to epilepsy comorbidity. <b>Results:</b> The <i>CACNA1A</i> rs16023 variant showed a significantly higher B allele frequency in DD/ID patients than in both control datasets and was also associated with epilepsy comorbidity. Genotype distribution analysis revealed that the BB genotype of rs16023 was more frequent in patients with epilepsy. <b>Conclusions:</b> The <i>CACNA1A</i> rs16023 variant may contribute to genetic susceptibility to DD/ID and epilepsy in Korean children, potentially through regulatory mechanisms. These findings support the relevance of calcium channel genes in neurodevelopmental disorders and highlight the importance of integrating functional annotation in variant prioritization.

<i>Background and Objectives:</i> Essential tremor (ET) is a common neurological disorder, typically presenting as bilateral, rhythmic, and symmetric kinetic or postural tremors. In contrast, Parkinson's disease (PD) is a progressive neurodegenerative disorder, characterized by resting tremor, rigidity, bradykinesia, and postural instability. Although both disorders involve tremor, ET and PD differ in clinical presentation and pathophysiology: ET generally involves action tremor and has a strong familial component, while PD more commonly presents with resting tremor and a weaker family history. A subset of ET patients may develop Parkinsonian features over time, although the relationship between ET and subsequent PD remains unclear. Genetic studies have identified only a few pathogenic variants in ET, suggesting it develops as a result of multifactorial genetic and environmental influences rather than simple Mendelian inheritance. ET is also recognized as a risk factor for developing PD, although the underlying mechanisms remain poorly understood. This study aimed to clarify potential genetic overlaps and distinctions in patients diagnosed with both ET and PD. <i>Materials and Methods:</i> We retrospectively analyzed 40 patients with a family history of ET or PD who were initially diagnosed with ET and later developed PD. Genetic screening and clinical assessments were conducted to investigate associated variants and clinical features. <i>Results:</i> Among these 40 patients, 17 different mutations were detected in 16 individuals. Three pathogenic or likely pathogenic variants were identified. The clinical characteristics and treatment responses of these patients were reviewed in relation to their genetic findings. Notably, none of the identified variants had previously been reported in association with PD following ET. <i>Conclusions:</i> A comprehensive clinical and genetic evaluation of ET patients who develop PD may offer insights into the underlying pathophysiology and inform future therapeutic strategies. Our findings support the need for further studies to explore the genetic landscape of patients with overlapping ET and PD features.

Major depressive disorder (MDD) is a prevalent mental illness that significantly impacts global health, with women showing twice the prevalence of men. This study employed the chronic social defeat stress (CSDS) model in female mice to investigate cellular and molecular changes in the prefrontal cortex (PFC) associated with depressive-like behaviors. Using single-nucleus RNA sequencing (snRNA-Seq), we examined transcriptomic alterations across various cell types in the PFC. Our results revealed that interneurons exhibited the most significant transcriptomic changes among all analyzed cell types. Notably, we found the Sox6⁺ interneurons (Sox6⁺Int) were enriched in the CSDS susceptible group. This enrichment was associated with enhanced inflammatory and immune responses, as well as alterations in synaptic function and mitochondrial pathways. Furthermore, we observed significant changes in cell-cell communication patterns, particularly between Sox6⁺Int and oligodendrocyte precursor cells (OPC). Weighted gene co-expression network analysis (WGCNA) identified several gene modules in Sox6⁺Int associated with specific depressive-like behaviors, implicating pathways related to inflammation, autophagy, and synaptic function. In particular, specific knockdown of Sox6 in neurons reversed the depressive-like behaviors. These findings provide novel insights into the cellular and molecular mechanisms underlying MDD in females, highlighting the potential role of Sox6⁺Int in stress-induced depression. Our study not only extends our understanding of the neurobiological basis of depression but also identifies potential therapeutic targets for sex-specific interventions in MDD treatment.

<h4>Introduction</h4>Previous studies have reported that IGF-1 single nucleotide polymorphism is associated with milk fat traits, but they are limited to trait association analysis. We previously identified a synonymous mutation c.258 A > G (rs322131043) in IGF-1, which influenced IGF-1 expression and caused differences in metabolism.<h4>Objectives</h4>This study aims to reveal a new regulatory function of IGF-1 c.258 A > G on milk fat metabolism.<h4>Methods</h4>Livers transcriptomics was used to identify differentially expressed genes between wild type mice (WT) and IGF-1 c.258 A > G mice (Homozygous mutation, Ho). Subsequently, lipid phenotyping, followed by metabolomics of mammary glands was conducted to verify transcriptomic findings. Finally, the potential mechanisms underlying IGF-1 c.258 A > G-induced changes in milk fat metabolism were explored though integrated transcriptomics-metabolomics analysis and Western blot validation.<h4>Results</h4>IGF-1 c.258 A > G changed the expression of genes related to lipid metabolism in livers of 8-week-old mice, including a 10-fold ‌lipoprotein lipase (LPL) expression (P < 0.01) and ‌80-90 % downregulation of acyl-CoA thioesterase 3 (Acot3), enoyl-Coenzyme A delta isomerase 3 (Eci3), fatty acid synthase (FASN), and sterol regulatory element binding protein1 (SREBP1) expression (P < 0.01). The milk fat content of Ho dams on the second day of lactation (L2D) was decreased 50 % than that of WT dams (P < 0.05), although there was no significant difference in adipose tissue of 8-week-old WT/Ho mice. The levels of triglycerides, sphingolipids and their related fatty acyl chains (10:0, 26:0, 14:2, 20:4, 11:3, 19:0) in mammary glands of L2D Ho dams were reduced 10-50 % observed by lipid metabolomics. And combined with transcriptomics and Western blot, the data suggested that a ‌2.5-fold upregulation of LPL expression‌ (P < 0.05) may contribute to the milk fat metabolism changes mediated by the ‌ IGF-1 c.258 A > G.<h4>Conclusion</h4>This study revealed new function of IGF-1 c.258 A > G on milk fat metabolism, thereby informing the development of targeted genetic breeding on milk fat trait.

Weaning is challenging for dairy calves, frequently resulting in digestive issues. This highlights the importance of implementing appropriate nutritional strategies to enhance gut health and support optimal growth. Postbiotics is a promising alternative to traditional probiotics, conferring health benefits without the risks associated with live bacteria. This study aimed to investigate the effect of dietary supplementation with a postbiotic from dead-cell <i>Limosilactobacillus ingluviei</i> C37 (postbiotic LIC37) on blood biochemical parameters and jejunal epithelium transcriptomic profiles in calves. Fourteen Holstein bull calves were randomly allocated into two groups (n = 7). The control group (CON) received a basic diet, while the postbiotic group (DCLI) was supplemented with 1 g/d of postbiotic LIC37 for 90 days. Blood samples were collected on days 76, 83, and 90, respectively. The jejunal epithelial tissue was obtained from four randomly selected calves per group at day 90 for transcriptome analysis. The results showed that postbiotic LIC37 supplementation reduced globulin, total protein, neutrophil (Neu) levels, and neutrophil-to-lymphocyte ratio (NLR) levels in the DCLI group (<i>p</i> < 0.05). Transcriptomic analysis identified 76 differentially expressed genes (DEGs), with significant upregulation of genes involved in fatty acid metabolism (FABP1), intestinal barrier function (B4GALNT2), and detoxification (GSTA1), alongside downregulation of immune response regulation (FCRLA, FCRL4). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses highlighted enrichment in pathways related to glutathione metabolism, drug metabolism, and vitamin digestion, indicating that postbiotic supplementation improved detoxification, oxidative stress defense, and nutrient absorption in calves. This study provides novel insights into the molecular mechanisms underlying the benefits of postbiotic LIC37 and supports its potential as a sustainable alternative to probiotics in calf nutrition.

Skeletal muscle is one of the largest tissues in the body. It is of great significance to analyze the molecular mechanism of skeletal muscle development for the further study of meat quality improvement and muscle diseases. CircRNA has been reported to be involved in many biological processes, but further research is needed in skeletal muscle. In this study, we detected the authenticity, stability, and spatio-temporal expression characteristics of circHOMER1 and its effect on the proliferation, apoptosis, and differentiation of muscle cells, and analyzed its possible molecular mechanism. The results showed that circHOMER1 exists in the skeletal muscle of the Ningxiang pig, is more stable than linear RNA, and is significantly upregulated in adipose tissue and during the early growth of myoblasts. In terms of function, overexpression of circHOMER1 significantly promoted the expression levels of proliferation marker genes and proteins and significantly increased the EdU positive cell rate, optical density (OD) value (at 450 nm), and proportion of S-phase cells. Overexpression of circHOMER1 also significantly promoted the expression levels of apoptosis marker genes and proteins and significantly increased the proportions of cells in Q2 (with late apoptosis) and Q3 (with early apoptosis). Overexpression of circHOMER1 significantly inhibited the expression levels of differentiation marker genes and proteins, significantly inhibited the differentiation index, and decreased the proportion of 5-nucleus muscle fibers. Conversely, opposite results were obtained after circHOMER1 interference. In terms of molecules mechanism, subcellular localization analysis showed that circHOMER1 was mainly distributed in cytoplasm, and mechanism analysis showed that circHOMER1 participated in myoblast development by forming a 4-element interaction network with 4 miRNAs, 2 lncRNAs, and 20 mRNAs, and possibly regulated myoblast development by encoding 79 amino acids. To sum up, we verified that circHOMER1 promoted the proliferation and apoptosis of myoblasts and inhibited their differentiation. It may regulate the development of myoblasts through ceRNA or by encoding small peptides. These results provided a reference for the regulation mechanism of muscle development and the breeding of Ningxiang pigs.

<h4>Background</h4>Characterized by microcirculatory disorder and oxidative stress microenvironment, the repair of bone defect after hip preservation therapy (such as core decompression) for osteonecrosis of the femoral head (ONFH) remains a clinical challenge. Thus, an ideal bone scaffold for treating ONFH should not only promote bone and vessel formation but also alleviate hypoxia and oxidative stress.<h4>Method</h4>We integrated manganese oxides (MnO_x) nanoparticles (NPs) with a 3D-printed poly(lactic-co-glycolic acid) (PLGA) scaffold to achieve this goal. The MnO_x NPs were synthesized using an oxidation reaction and the scaffold was 3D-printed using a fused deposition modeling method. The characterization and the enzyme-like activity of the scaffold was investigated. The biocompatibility and biofunctions of the scaffold were evaluated both in vitro and in vivo, including the antioxidant capacity, the effects on promoting bone formation and vascularization, and the therapeutic effect in animal model.<h4>Results</h4>The resultant MnO_x-doped PLGA scaffold could catalyze reactive oxygen species into oxygen through its superoxide dismutase (SOD)-like and catalase (CAT)-like activities. In vitro studies revealed that this multienzyme-like activity of the scaffold could be maintained for more than 30 days, thereby improving cell viability under oxidative stress. The underlying mechanism was shown to involve regulation of the antioxidant activity of cells via PI3K/AKT signaling pathway. The scaffold also significantly improved capabilities of osteogenesis and angiogenesis compared to pure PLGA scaffold. In vivo studies further demonstrated that the combination therapy of core decompression and scaffold implantation efficiently reduced osteoblast necrosis and enhanced vascularized bone formation in a clinically relevant ONFH rabbit model.<h4>Conclusion</h4>The 3D-printed MnO_x-doped PLGA scaffold not only relieve oxidative stress to protect osteocytes under ONFH microenvironment but also promote vascularized bone formation, showing the potential for treating ONFH.<h4>Translational potential of this article</h4>PLGA has been already applied in clinical bone implants. Mn is an essential trace element for the human body and MnO_x NPs offer the advantage of biocompatibility, ease of large-scale preparation, and low cost. Hence, this scaffold has the potential for clinical translation in the treatment of ONFH.

The maintenance of intestinal homeostasis relies on the continuous self-renewal and differentiation of intestinal stem cells (ISC). This study examines the effects of monochromatic light on the self-renewal and differentiation of ISC, thereby elucidating the mechanisms by which monochromatic light influences the growth and development of broilers. The post-hatching day 0 (P0) Cobb 500 broilers were randomly assigned to white light (WL), red light (RL), green light (GL) and blue light (BL) until P35. The findings indicated that the body weight of broilers in the BL group was significantly higher than that observed in the WL groups, and the BL group exhibited an increase in jejunal villus length, a decrease in crypt depth, and a reduction in the villus to crypt (V/C) ratio. The mRNA levels of Sglt1, Glut1, Slc7a5, Fabp2, sIgA, and Tff3 in the jejunum of the BL group were significantly increased relative to those in the other light groups. Furthermore, there was a notable increase in the number of enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. The mRNA levels of Muc2, Lyz, Alpi and Chga as well as the protein levels of SI, Lyz and Chga were also significantly raised. Additionally, the BL group promoted the expression of PCNA and Lgr5, increased the mRNA levels of Olfm4 and Ccnd1, and elevated the protein levels of Lrp6 and β-catenin, while decreasing the protein level of APC. Meanwhile, BL group resulted in increased mRNA levels of Notch1, Notch2 and Dll4, along with an increase in the protein level of Notch1. Relative to the WL group, the BL group also enhanced the depth of cecal crypt and the number of PCNA immunoreactive cells, as well as increased the mRNA levels of Muc2, Chga, Lgr5, sIgA and Tff3. These results suggest that BL may facilitate the self-renewal and differentiation of ISC and enhance the morphology and function of the intestinal epithelium by activating the Wnt/β-catenin and Notch signaling pathways.

<h4>Introduction</h4>Atrial fibrillation (AFib) is a sustained form of cardiac arrythmia that occurs due to sympathetic overdrive, neurohumoral and electrophysiological changes. Sympatho-renal modulatory approach via miRNA-based therapeutics is likely to be an important treatment option for AFib. The study was aimed to unravel the common miRNAs as therapeutic targets involved in sympatho- renovascular axis to combat AFib.<h4>Methods</h4>We employed the bioinformatics approach to discover differentially expressed genes (DEGs) from microarray gene expression datasets GSE41177 and GSE79768 of AFib patients. Concomitantly, genes associated with sympathetic cardio-renal axis, from Genetic Testing Registry (GTR) of National Center for Biotechnology Information (NCBI) were also analyzed. Overlapping miRNAs that target the maximum number of genes across all three pathological conditions perpetuating AFib were shortlisted. To confirm the reliability of the identified miRNAs, differential expression analysis was performed on miRNA expression profiles GSE190898, GSE68475, GSE70887 and GSE28954 derived from AFib patient samples.<h4>Results</h4>ShinyGO analysis revealed enrichment in beta-adrenergic signaling, calcium signaling, as well as G protein-coupled receptor (GPCR) signaling involved in post synaptic membrane potential. The intersection of top 10 modules in miRNA-mRNA network revealed hub miRNAs having highest node degree, maximum neighborhood component (MNC), and maximal clique centrality (MCC) scores. Differential expression analysis revealed hub miRNAs identified through integrated approach were found to be significantly dysregulated in AFib patients.<h4>Conclusion</h4>This integrated approach identified 6 hub miRNAs, 4 reported (miR-101-3p, miR-23-3p, miR-27-3p, miR-25-3p) and 2 novel (miR-32-5p, miR-92-3p) miRNAs that might act as putative biomarkers for AFib.

The cardiovascular and musculoskeletal systems are two core systems essential for maintaining human physiological functions and their dynamic interactions play a critical role in overall health. Exosomes, nanosized vesicles released by cells, contain bioactive substances including microRNA, long non‑coding RNA, lipids and proteins and participate in the pathophysiological regulation of multiple organ systems by mediating intercellular communication. Bone‑derived exosomes ameliorate cardiovascular diseases through the regulation of oxidative stress, inflammatory responses and apoptosis. Conversely, cardiovascular‑derived exosomes enhance bone homeostasis by suppressing osteoclast activity or promoting osteogenic differentiation, but they may also exacerbate pathological progression in conditions such as osteoarthritis. Skeletal muscle‑derived exosomes protect cardiomyocytes in muscular dystrophy through functional molecules delivery. However, under pathological conditions such as sarcopenia, skeletal muscle‑derived exosomes may aggravate cardiac dysfunction by activating pro‑apoptotic signals. Similarly, cardiovascular‑derived exosomes exhibit dual roles in skeletal muscle regulation, promoting regeneration while potentially inducing atrophy during heart failure. In addition, exosomes demonstrate significant clinical value as diagnostic biomarkers and targeted drug delivery vehicles, both for early disease detection and regenerative therapies. The present review systematically outlined the mechanisms underlying exosome‑mediated bidirectional crosstalk between the cardiovascular and musculoskeletal systems and explores their clinical application potential. It provided theoretical insights and novel perspectives for further research into the pathogenesis and therapeutic strategies of cardiovascular and musculoskeletal diseases.

Allogeneic stem cell transplantation (SCT) remains the only curative therapy for patients with high-risk myelodysplastic syndromes (MDS). Due to age, comorbidities, or lack of urgency, many receive only palliative therapies to improve quality of life. Some patients remain untreated due to a lack of symptoms or low progression risk. Data on the impact of palliative therapies on overall survival (OS) and leukemia-free survival (LFS) are limited. Using the Düsseldorf MDS Registry, we compared outcomes of patients receiving red blood cell transfusions (RBCT) alone to the outcome of patients receiving RBCT combined with iron chelation therapy (ICT), erythropoietin (EPO), antithymoglobulin (ATG), or lenalidomide (LENA). Matched-pairs analysis was conducted using age, gender, and prognostic scores (Revised International Prognostic Scoring System or Chronic Myelomonocytic Leukemia-specific Prognostic Scoring System). ICT-treated patients (n = 85) had significantly improved OS (70 vs. 21 months, p < 0.001) and lower 5-year AML progression (3.5% vs. 28.2%, p < 0.001). Similar benefits were seen with EPO (n = 210; OS: 63 vs. 24 months, p < 0.001; AML: 5.7% vs. 19%, p = 0.007) and LENA (n = 30; OS: 92 vs. 57 months, p = 0.049; AML: 0% vs. 16.7%, p = 0.024). ATG (n = 11) showed no significant improvement in OS (79 vs. 64 months) or AML progression (0% vs. 18.2%). While recognizing the limitations of matched-pairs analysis versus randomized trials, our findings indicate a survival benefit from ICT, EPO, or LENA versus RBCT alone. The year of diagnosis did not independently affect OS or LFS. These results support the use of selected palliative therapies to improve long-term outcomes in MDS patients. KEY POINTS: Treating patients with myelodysplastic syndromes with non-curative therapies beyond red blood cell transfusions like iron chelation therapy, erythropoietin, or lenalidomide has a positive impact on overall survival and leukemia-free survival.

<h4>Introduction</h4>Keloids are pathologic scars that result from abnormal wound healing processes following skin trauma, with a higher prevalence in the Black population. The pathogenesis of keloids is not fully understood, hindering effective treatment options for this highly disfiguring and distressing condition.<h4>Methods</h4>Biopsies from lesional and non-lesional skin from keloid patients and healthy skin from age/gender/race-matched controls were collected and analyzed using RNA-sequencing, RT-qPCR, and immunohistochemistry. Spearman analysis was used to evaluate the correlations between biomarker expressions and clinical severity measurements.<h4>Results</h4>Both keloidal lesions and non-lesional skin showed a distinct transcriptomic profile compared to healthy skin. Keloids demonstrated significant upregulation of fibrosis-related markers (e.g., COL10A1, COL11A1, and BMP1). Lesional and/or non-lesional samples showed significant upregulation of key immune biomarkers belonging to T-cells (e.g., CD2, CD3D, and CD3E), T-cell/NK-cell activation/migration (e.g., CCL19, CCR7, GZMA, GZMB, and GZMK), Th1 (e.g., OASL, MX1, CCL4), Th2 (e.g., IL4R, OX40/TNFRSF4, and OX40L/TNFSF4), and Th17/22 (e.g., S100A7, S100A8, S100A9, and CCL20). Multiple immune biomarkers expression (e.g., CCL2, CXCL1, and S100A7) in lesional and/or non-lesional skin significantly and positively correlated with keloid severity parameters (e.g., keloid size, distensibility, and number).<h4>Conclusion</h4>Both lesional and non-lesional keloid skin show distinct upregulation of immune axes, underscoring the role of inflammation in keloid pathogenesis and pointing to potential novel therapeutic targets.

Vulnerability to climate hazards and infectious diseases is not gender-neutral, meaning that men, women, and other gender identities experience different risks. Leptospirosis is a zoonotic climate-sensitive infectious disease caused by the bacteria Leptospira and is transmitted to humans through contact with infected animals or contaminated environments, particularly soil and floodwater. Globally, studies report a higher risk of leptospiral infection among men than women, a trend also observed in Salvador, Brazil; however, the factors driving this difference are poorly understood. This study aimed to investigate how Leptospira exposure differs between men and women living in urban informal settlements in Salvador. We conducted a cross-sectional serosurvey among 761 adults (280 men and 481 women) in four communities previously identified as high-risk by surveillance data. Using a causal inference approach and a two-part sex-disaggregated analysis, we applied logistic regression models to examine: (1) the association between perceived severity and high-risk behaviours with Leptospira seropositivity, and (2) the association between perceived severity with high-risk behaviours. Seroprevalence was 14.6% (95% CI: 10.5%-18.8%) in men and 9.4% (95% CI: 6.7%-12.0%) in women. Men who perceived leptospirosis as extremely serious had lower odds of being seropositive (OR: 0.38, 95% CI: 0.15-0.99), walking through sewage (OR: 0.41, 95% CI: 0.17-1.00), and walking barefoot outside (OR: 0.24, 95% CI: 0.08-0.76) compared with men who perceived leptospirosis as less serious. These associations were not observed in women and differed across age groups in men. Behaviours were not associated with seropositivity in either gender. Our results identify perceived severity as a potential driver of high-risk behaviours and exposure in men, indicating perceptions as targets for health promotion programs, while also highlighting evidence gaps in understanding exposure risks among women. As the first sex-disaggregated study investigating Leptospira exposure risks, we advocate for a gendered lens in future studies to understand gender-specific risks.

<i>N</i>⁶-Methyladenosine (m⁶A) modification plays crucial roles in tissue development and homeostasis. However, the mechanisms underlying cellular adaptation of m⁶A modification and their impact on protein synthesis machinery remain unclear. VIRMA, the largest and evolutionarily conserved core of the m⁶A methyltransferase complex, is highly expressed in the embryonic brain and various cancers. Here, we demonstrate that VIRMA-mediated m⁶A modification is essential for active ribosome biogenesis. VIRMA depletion destabilizes the entire writer complex and reduces m⁶A levels, leading to decreased proliferation and increased apoptosis of neural progenitor/stem cells, ultimately causing severe forebrain developmental defects. Mechanistically, VIRMA depletion impairs ribosome biogenesis by inhibiting mRNA decay, triggering a p53-dependent stress response and compromising global protein synthesis. These findings extend to some cancer cells, suggesting a potential conservation of this mechanism. Overall, our study reveals the critical role of m⁶A in adapting protein synthesis machinery during brain development.

The transcription factors TFEB and TFE3 modulate expression of lysosomal, autophagic, and metabolic genes to restore energy and cellular homeostasis in response to a variety of stress conditions. Since their role during vertebrate development is less characterized, we used CRISPR/Cas9 to deplete tfeb, tfe3a, and tfe3b in zebrafish. The simultaneous lack of these genes compromised embryo survival during early development, with an almost complete lethality of the larvae by 8-10 dpf. The knockout animals showed apoptosis in brain and retina and alterations in pancreas, liver, and gut. Exocrine pancreas presented the most severe defects, with accumulation of abnormal zymogen granules leading to acinar atrophy in embryos and pancreatitis-like phenotypes in adults; likely due to a block of the autophagy machinery implicated in removal of damaged granules. Knockout animals displayed increased susceptibility to oxidative and heat-shock stress. Our work reveals an essential role of Tfeb and Tfe3 in maintaining cellular and tissue homeostasis during development.

We studied the characteristics and survival of patients with sorafenib-treated HCC and impact of underlying etiology on outcomes. This retrospective multicenter study recruited patients with sorafenib-treated advanced HCC (12/2016 to 4/2023) till death or the study end (2/2024). Time to progression (TTP) and overall survival (OS) were recorded. We evaluated; Clinico-laboratory and imaging predictors of OS, The impact of underlying etiology on tumor variables, outcomes and tolerance for sorafenib > 6 months. This study included 706 patients. Median duration of Sorafenib therapy was 240.00 (90.00-360.00) days. Median OS was 314.00(146.00-601.00) days. Median TTP was 180.00(90.00-330.00) days. COX regression revealed that the independent factors of mortality were baseline AST, Tumor size, hepatic vein thrombosis (HVT), development of jaundice and shifting to Regorafenib. Advanced HCCs were more common on top of non-cirrhotic non-viral and HBV-related liver disease. Adverse events, TTP and tumor response didn't differ with the underlying etiology. Median OS was lower in non-viral-related HCC than HCV-related HCC (218.00 versus 326.50 days, P-value = 0.048). Patients who continued sorafenib > 6 months had lower AFP, HVT, adverse effects and better tumor response after 3 months. OS is lower in non-viral Sorafenib-treated HCC compared with viral-related HCC and Sorafenib was well-tolerated among different HCC etiologies.

Interleukin-17 (IL-17)-producing γδ T (Tγδ17) cells are innate-like mediators of intestinal barrier immunity. Although IL-17-producing helper T cell and group 3 innate lymphoid cell plasticity have been extensively studied, the mechanisms governing Tγδ17 cell effector flexibility remain undefined. Here, we combined type 3 fate mapping with single-cell ATAC-sequencing/RNA-sequencing multiome profiling to define the cellular features and regulatory networks underlying Tγδ17 cell plasticity. During homeostasis, Tγδ17 cell effector identity was stable across tissues, including for intestinal T-bet⁺ Tγδ17 cells that restrained interferon-γ production. However, Salmonella enterica subsp. enterica serovar Typhimurium infection induced intestinal V_γ6⁺ Tγδ17 cell conversion into type 1 effectors, with loss of IL-17A production and partial RORγt downregulation. Multiome analysis revealed a trajectory along V_γ6⁺ Tγδ17 cell effector conversion, with TIM-3 marking ex-Tγδ17 cells with enhanced type 1 functionality. Last, we characterized and validated a critical AP-1 regulatory axis centered around JUNB and FOSL2 that controls V_γ6⁺ Tγδ17 cell plasticity by stabilizing type 3 identity and restricting type 1 effector conversion.

CD8⁺ T cell exhaustion (T_ex) limits immune control of cancer, but the underlying molecular drivers are unclear. In the present study, we identified the prostaglandin I₂ (prostacyclin) receptor PTGIR as a cell-intrinsic regulator of T cell exhaustion. Transcriptomic profiling of terminally exhausted (T_t_ex) CD8⁺ T cells revealed increased activation of the nuclear factor erythroid 2-related factor 2 (NRF2) oxidative stress response pathway. Enhancing NRF2 activity (by conditional deletion of Kelch-like ECH-associated protein 1 (KEAP1)) boosts glutathione production in CD8⁺ T cells but accelerates terminal exhaustion. NRF2 upregulates PTGIR expression in CD8⁺ T cells. Silencing PTGIR expression enhances T cell effector function (that is, interferon-γ and granzyme production) and limits T_tex cell development in chronic infection and cancer models. Mechanistically, PTGIR signaling impairs T cell metabolism and cytokine production while inducing transcriptional features of T_ex cells. These findings identify PTGIR as a NRF2-dependent immune checkpoint that regulates balance between effector and exhausted CD8⁺ T cell states.

Developmental origins and their associations with lineage plasticity and treatment response in B-cell progenitor acute lymphoblastic leukemia (B-ALL) are mostly unexplored. Here, we integrated single-cell transcriptome sequencing (scRNA-seq) of 89 B-ALL samples with a single-cell atlas of normal human B cell development incorporating functional and molecular assays. We observed subtype- and sample-dependent correlation with normal developmental stage, with intra-subtype and intra-patient heterogeneity. We show that subtypes prone to shift from the B-lineage (for example BCR::ABL1, KMT2A-R and DUX4-R B-ALL) are enriched for multipotent progenitors and show this developmental stage exhibits CEBPA activation and retains myeloid potential, providing a mechanistic explanation for this clinical observation. We developed a 'multipotency score' most enriched in subtypes exhibiting lineage plasticity that was independently associated with inferior survival. Thus, multipotent B-ALL states reflect the early progenitor origins of a subset of patients with B-ALL and may be relevant for understanding lineage shifting following conventional chemotherapy or immunotherapies.

<h4>Objective</h4>Epigenetic modifications including histone post translational modifications can influence gene expression in adipocytes, potentially contributing to metabolic dysfunctions, obesity, and insulin resistance. Despite recent advances in the characterization of the mouse adipocyte epigenome, epigenetic characterization of adipocytes in vivo has been challenging, particularly across different adipose depots and of several epigenetic modifications.<h4>Methods</h4>Here, we use specific reporter mice labelling brown, beige and white adipocytes, diphtheria toxin-mediated ablation of beige adipocytes, and Cleavage Under Targets and Tagmentation (CUT&Tag) to generate paired single mouse datasets of five histone marks. We perform an integrative multi-omics factor analysis (MOFA) of H3K4me3, H3K27me3, H3K4me1, H3K27ac and H3K9me3 in brown, white and beige adipocytes from three distinct mouse adipose tissue depots obtained during cold exposure and thermoneutrality.<h4>Results</h4>Our analysis reveals that enhancers distinguish adipocytes by their tissue of origin, with H3K4me1 deposition differentiating between beige and brown adipocytes. Beige adipocytes poised promoters associated to thermogenic genes during warming. Diphtheria toxin-mediated ablation of beige adipocytes shows that non-beigeing white adipocytes in inguinal adipose tissue and beige adipocytes are not inherently epigenetically different suggesting that they share a common developmental progenitor.<h4>Conclusions</h4>These paired multimodal data comprise an extensive resource (https://github.com/vonMeyennLab/mAT_CE_Atlas) for the further exploration of the mouse adipocyte epigenome which will enable discovery of regulatory elements governing adipocyte identity and gene regulation.

Marine-derived antioxidant astaxanthin (AST), a potent antioxidant carotenoid, has gained significant attention for its potential therapeutic effects in various diseases, including cancer and metabolic disorders. When combined with exercise, which is a well-established intervention for improving health, these two modalities may offer synergistic benefits that extend beyond their individual effects. This review explores the molecular mechanisms underlying the synergistic therapeutic effects of AST and exercise in cancer, metabolic diseases, and healthy individuals. AST exerts its beneficial effects primarily through its ability to reduce oxidative stress, modulate inflammation, and enhance cellular signaling pathways, including those involved in apoptosis, autophagy, and mitochondrial function. It has been shown to suppress tumor growth, improve insulin sensitivity, and protect against the adverse effects of chronic diseases, such as cardiovascular complications and neurodegenerative conditions. Similarly, exercise induces a wide array of molecular adaptations, including the activation of key metabolic pathways, enhancement of mitochondrial biogenesis, and modulation of inflammatory responses. These effects improve metabolic health, reduce cancer risk, and promote overall well-being. The combination of AST supplementation and exercise may provide a more potent therapeutic strategy, targeting multiple molecular pathways simultaneously. This synergy may not only enhance the effectiveness of each intervention but also reduce the side effects commonly associated with pharmacological treatments. This review discussed the current evidence for the synergistic effects of AST and exercise, highlighted the molecular mechanisms involved, and suggested potential clinical applications for these interventions in cancer, metabolic diseases, and healthy populations.

Oxetanes are 4-membered cyclic monoethers which have found important applications in medicinal chemistry as polar and metabolically stable isosteric replacements for <i>gem</i>-dimethyl and carbonyl groups. This work reviews possible synthetic strategies towards these strained heterocycles, covering both de novo constructions of the 4-membered ring as well as derivatisations of oxetane building blocks, then reactivity of oxetanes in terms of ring-opening and ring-expansion reactions, and finally total syntheses of selected oxetane-containing natural products. The literature review primarily covers reports made after the year 2015, but a few older contributions that were considered relevant are also discussed.

Breast cancer continues to be one of the most perilous diseases globally due to the challenges in identifying cost-effective and targeted targets for effective treatment strategies. In response to these unmet demands, much research has focused on investigating the anti-breast cancer properties of natural compounds due to their multi-target modes of action and favorable safety profiles. Numerous extracts of medicinal plants, essential oils, and natural bioactive substances have exhibited anticancer properties in preclinical breast cancer models. Nonetheless, the clinical utilization of therapies based on natural chemicals is constrained by challenges such as inadequate solubility and permeability. Innovative drug delivery systems utilizing nanoparticles are currently being investigated as adaptable solutions to overcome these limitations. Polysaccharide-based nanoparticles exhibit promising biodegradability and biocompatibility, making them suitable for targeted drug delivery systems among the diverse nanocarrier types. Many polysaccharide nanocarriers, including chitosan, hyaluronic acid, cellulose, starch, and complex polysaccharides, have been shown to enhance the bioavailability and therapeutic effectiveness of phytocompounds. This review examines the anticancer properties of phytocompounds and polysaccharide nanocarriers in breast cancer before focusing on the use of polysaccharide-based nanocarriers to deliver phytocompounds for the treatment of breast cancer.

Myeloid/lymphoid neoplasms with eosinophilia and tyrosine kinase gene fusions (MLN-TK) represent rare hematological malignancies driven by pathological fusion genes involving tyrosine kinase genes. Among these, rearrangements of the <i>PDGFRB</i> gene, particularly the <i>ETV6::PDGFRB</i> rearrangement, are frequently observed as pathogenic mutations. Conversely, instances of the <i>MPRIP::PDGFRB</i> fusion gene are rarely documented. In this case report, we present a 32-year-old previously healthy Thai male who presented to the hospital with constitutional symptoms and marked splenomegaly. His complete blood count revealed mild anemia, marked leukocytosis with hypereosinophilia, and mild thrombocytopenia. A bone marrow study showed hypercellular marrow with granulocytic hyperplasia extensively involved with eosinophils, without morphological evidence of blasts. Conventional cytogenetics identified a t (5; 17) (q33; p13). Further targeted RNA analysis using next-generation sequencing (NGS) detected a fusion gene involving <i>MPRIP::PDGFRB</i>. The patient was diagnosed with myeloid/lymphoid neoplasms with eosinophilia and <i>MPRIP::PDGFRB</i> rearrangement in the chronic-phase disease and was initiated on oral imatinib at a daily dose of 100 mg. One month after initiating the treatment, the patient achieved a hematological response consistent with complete response (CR) criteria. Imatinib therapy has been well-tolerated without reported adverse events, and a 1-year molecular assessment confirmed the achievement of complete molecular response (CMR).

It has been well established that selenium (Se) should be provided as a supplement to grazing cattle in regions where the soils and forages are deficient in this trace mineral [...].

<b>Background</b>: Head and neck cancer (HNC) remains a global health challenge with a poor 5-year survival rate among patients with relapsed or advanced-stage disease. Immune checkpoint blockade therapies have emerged as a promising approach to improve outcomes; however, their effectiveness is limited, with response rates of only 15-20% because of immune evasion mechanisms. MicroRNA (miRNA) dysregulation plays a key role in facilitating such immune evasion. In this study, we aim to identify specific miRNAs whose altered expression contributes to immune escape in HNC. <b>Methods</b>: We employed an integrated bioinformatics approach, incorporating differential expression analysis, survival analysis, target prediction, KEGG immune pathway analysis, a protein-protein interaction network, and the identification of hub genes using in silico tools. <b>Results</b>: Our analysis revealed that a high expression of miR-18a and miR-2355 was associated with reduced survival, with the median survival decreasing from 42.9 to 27.8 months, respectively, in advanced-stage patients. Conversely, a low expression of let-7c and miR-6510 was linked to poor prognosis, with survival decreasing from 40.1 to 19.2 months and from 50.1 to 26.8 months, respectively, across disease progression. Further pathway analysis revealed that these miRNAs are significantly involved in the regulation of key immune evasion signaling pathways, including T cell receptor, PD-L1/PD-1 checkpoint, JAK-STAT, TGF-beta, NF-kappa B, and TNF signaling pathways. Hub gene analysis identified AKT1, STAT3, NFKB1, CD4, IL2RB, TLR4, and CTLA-4 as potential dysregulated miRNA targets, with enrichment in immune-related signaling pathways. <b>Conclusions</b>: Taken together, these findings suggest that targeting these miRNAs could modulate immune evasion mechanisms and potentially enhance the efficacy of ICB therapies in HNC.

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are linked by shared genetic mutations and overlapping clinical features, forming a clinical spectrum. This systematic review and meta-analysis analysed 97 studies, including 3212 patients with key ALS/FTD gene mutations, to identify gene-specific behavioural profiles. <i>Chromosome 9 open reading frame 72</i> (<i>C9orf72</i>) mutations were strongly associated with psychotic symptoms and aggression, while <i>superoxide dismutase 1</i> (<i>SOD1</i>) mutations had minimal cognitive effects. <i>Progranulin</i> (<i>PGRN</i>) mutations correlated with apathy and hallucinations, <i>microtubule-associated protein tau</i> (<i>MAPT</i>) <i>mutations</i> with disinhibition, and <i>charged multivesicular body protein 2B</i> (<i>CHMP2B</i>) with social impairments. <i>Fused in sarcoma</i> (<i>FUS</i>) mutations caused early sleep disturbances, <i>TANK-binding kinase 1</i> (<i>TBK1</i>) led to disinhibition, and <i>presenilin 1 and 2</i> (<i>PSEN1/2</i>) was linked to severe aggression. Prodromal cognitive changes in <i>PGRN</i>, <i>MAPT</i>, and <i>CHMP2B</i> mutations suggested early disease onset. Despite overlapping symptoms and clinical heterogeneity, understanding gene-specific patterns could inform tailored care strategies to enhance the quality of life for ALS and FTD patients. This study calls for refined guidelines integrating genetic behavioural profiles to improve patient and family support.

High-grade gliomas (HGGs, WHO grades 3-4) are highly aggressive, with a poor prognosis and treatment resistance. Immune evasion may contribute to their progression, but the role of cytotoxic T-lymphocyte immune evasion (CTLE) is not well-validated. This study analyzed the transcriptomic data of 525 patients from TCGA-GBM-HG_U133A. Two molecular subtypes were identified based on 182 CTLE-associated genes, with 238 differentially expressed genes between them. A prognostic model was developed, identifying BST2 and DIRAS3 as key risk factors, and validated in multiple cohorts. The subtypes had distinct immune profiles, with Cluster 2 showing higher immune infiltration but a poorer prognosis. The model had a good predictive performance. High-risk patients had upregulated BST2 and DIRAS3, linked to immunosuppression and shorter survival. Knockdown experiments confirmed their roles in GBM cell migration and invasion. Mechanistically, they promote immune evasion. BST2 and DIRAS3 could be therapeutic targets for HGG immunotherapy.

Male infertility is an under-recognized global health burden. Accumulating evidence position the intestinal microbiota as a pivotal regulator of testicular function, underpinning the emerging gut microbiota-testis axis. This narrative review introduces the conceptual term "androbactome", referring to gut microorganisms and microbial genes that are hypothesized to influence androgen biosynthesis, spermatogenesis, and broader reproductive endocrinology. The documented worldwide decline in sperm concentration heightens the urgency of clarifying microbe-mediated influences on male reproductive capacity. The synthesis of preclinical and clinical findings reveals four principal pathways by which dysbiosis compromises fertility: systemic inflammation, oxidative stress, endocrine disruption, and epigenetic alteration. Lipopolysaccharide-driven cytokinaemia, reactive oxygen species generation, hypothalamic-pituitary-gonadal axis suppression, and aberrant germ cell methylation collectively impair sperm quality and hormonal balance. Short-chain fatty acids, secondary bile acids, and indole derivatives emerge as pivotal messengers within this crosstalk. Therapeutic approaches targeting the androbactome, namely dietary optimization, probiotic or prebiotic supplementation, and fecal microbiota transplantation, have demonstrated encouraging improvements in sperm parameters and testosterone levels, yet the causal inference is constrained by predominantly cross-sectional designs and limited long-term safety data. Recognizing the androbactome as a modifiable determinant of male fertility may open new avenues for personalized diagnosis, risk stratification, and adjunctive therapy in regard to idiopathic infertility. The integration of multi-omics platforms to characterize microbial and metabolomic signatures promises to enrich diagnostic algorithms and guide precision interventions, but rigorously controlled longitudinal and interventional studies are required to secure a translational impact.

In addition to the male genome, the fertilizing spermatozoon delivers to the oocyte several factors whose deficiency can cause embryo dysfunction. Sperm oocyte-activating factor, identified as phoshoplipase C zeta (PLCζ), drives oocyte exit from meiotic arrest through a signaling pathway initiated by periodic rises of free cytosolic Ca²⁺ concentration (calcium oscillations). Sperm centrioles, together with oocyte proteins, form centrosomes that are responsible for aster formation, pronuclear migration, and DNA polarization before nuclear syngamy and subsequent mitotic divisions. Sperm DNA fragmentation can be at the origin of aneuploidies, while epigenetic issues, mainly abnormal methylation of DNA-associated histones, cause asynchronies of zygotic gene activation among embryonic cells. Sperm long and short non-coding RNAs are important epigenetic regulators affecting critical developmental processes. Dysfunction of sperm PLCζ, centrioles, DNA, and RNA mostly converge to aneuploidy, developmental arrest, implantation failure, miscarriage, abortion, or offspring disease. With the exception of DNA fragmentation, the other sperm issues are more difficult to diagnose. Specific tests, including heterologous human intracytoplasmic sperm injection (ICSI) into animal oocytes, genetic testing for mutations in <i>PLCZ1</i> (the gene coding for PLCζ in humans) and associated genes, and next-generation sequencing of sperm transcriptome, are currently available. Oral antioxidant treatment and in vitro selection of healthy spermatozoa can be used in cases of sperm DNA fragmentation, while ICSI with assisted oocyte activation is useful to overcome oocyte-activation defects. No clinically confirmed therapy is yet available for sperm RNA issues.

Huntington's disease (HD) is the most common autosomal dominant neurodegenerative disorder, characterized by a triad of motor dysfunction, cognitive decline, and psychiatric disturbances. While recent efforts have focused on developing disease-modifying therapies, no treatment has yet demonstrated clinical efficacy. As a result, symptomatic treatment remains the cornerstone of care. However, high-quality evidence from large randomized trials is limited, and therapeutic decisions must rely on clinical expertise and extrapolation from other neurological or psychiatric conditions. This narrative review provides a comprehensive and practical overview of symptomatic treatment strategies for HD with emphasis on the pathophysiological underpinnings of each symptom and the molecular mechanisms of available and emerging therapies, aiming to support rational, individualized management. Finally, we highlight the critical role of non-pharmacological interventions and the need for multidisciplinary approaches to optimize patient outcomes and quality of life.

Chronic hepatitis B virus (HBV) infection has been implicated in the development of liver diseases, such as hepatitis, fibrosis, cirrhosis, and cancer, which negatively affect the patients' quality of life and impacts a high economic strain on patients. The persistence of covalently closed circular DNA (cccDNA) allows the propagation of the infection, and no drug have been developed to completely eliminate cccDNA. The available drugs for chronic hepatitis B (CHB) are classified into nucleos(t)ide analogs (NAs) and interferon-α (IFN-α)/pegylated interferon α (Peg-IFN-α). However, these treatments do not effectively eradicate hepatitis B surface antigen (HBsAg) and their clinical efficacy is limited. The potential of IFN-based clinical cure is increasingly attracting interest from hepatologists, but the therapeutic outcomes of this intervention are suboptimal and some of them are associated with various complications. Although several novel antiviral drugs are being investigated, however, achieving a clinical cure based on monotherapy is currently challenging. The efficacy of IFN therapy is influenced by host and viral factors. This article provides a comprehensive review of host-related factors that affect the IFN therapy for CHB. A thorough understanding and management of these host-related factors will enhance the efficacy of interferon treatment, minimize adverse reactions, improve patient tolerance, and thereby increasing the clinical cure rate of hepatitis B.

<h4>Objective</h4>Childhood obesity has become a global public health crisis. Previous studies have shown that nutritional supplementation during pregnancy may be protective against offspring obesity. However, the research in this area is still emerging and the impact of moderators, such as birth weight, upon outcomes has not been fully explored. This study aimed to examine the combined effect of maternal supplementation with iron, calcium, folic acid, and multivitamin during pregnancy on the risk of obesity in Chinese preschoolers born macrosomia.<h4>Methods</h4>A total of 6,031 singleton children, born macrosomia, aged 3-6.5 years old were recruited from Longhua District in Shenzhen of China in 2021. Their mothers were asked to complete a structured questionnaire for collecting the sociodemographic characteristics of the child and parents, the child's birth-related characteristics, and maternal supplementation with iron, calcium, folic acid, and multivitamins during pregnancy. The children's weight and height were measured using a standardized method by well-trained medical staff from the Women's and Children's Hospital of Longhua District of Shenzhen.<h4>Results</h4>After controlling for confounding variables, including other nutrients, the results of a series of logistic regressions showed that only iron supplementation (AOR = 0.75, 95% CI = 0.60-0.92) during pregnancy was negatively associated with the presence of obesity in preschoolers born macrosomia in boys. In contrast, there was no independent associations between maternal prenatal ingestion of iron, calcium, folic acid, or multivitamin supplements and obesity in preschool girls born macrosomia. Examination of interaction effects through crossover analyses showed that maternal supplementation with both iron and calcium (AOR = 0.68, 95% CI = 0.49-0.94), and both iron and multivitamins (AOR = 0.64, 95% CI = 0.48-0.86) during pregnancy significantly reduced the risk of obesity in male preschoolers born macrosomia. Furthermore, interaction analysis found a multiplicative interaction between maternal iron and multivitamin supplementation during pregnancy on the risk of obesity in male preschoolers born macrosomia (IOR = 0.52, 95% CI = 0.35-0.79).<h4>Conclusion</h4>Our findings suggest that iron supplementation during pregnancy may reduce the risk of obesity in preschoolers born macrosomia in boys, with this effect enhanced with the combined ingestion of calcium and multivitamin supplementation.

No abstract available.

Current LNP technology faces challenges that must be addressed to enhance the functionality of mRNA therapeutics. Recent studies show disulfide-conjugated molecules improve cell membrane permeability. Here, we investigated incorporating cyclic disulfide (CDL) units into lipid components of LNPs to enhance LNP-mRNA performance. A lipid library with branched and unbranched alkyl chains (C16-C20) and tertiary amine groups modified with CDLs was designed. While cellular uptake was unchanged, some mRNA-loaded LNPs with CDLs achieved more than 2-fold higher transfection efficiency than LNPs with MC3 or SM102 alone. Intracellular analysis revealed that the addition of CDL lipids significantly promoted endosomal escape. The CDL-incorporated LNPs administered subcutaneously in mice showed significantly higher luciferase gene expression compared to LNPs without CDL. Additionally, LNPs encapsulating OVA antigen-encoding mRNA induced a potent antitumor response against the EG7-OVA lymphoma model. These results suggest CDL modifications enhance LNP-based mRNA delivery, offering potential for broader therapeutic applications and improved clinical outcomes.

In the present study, five novel dexketoprofen amide derivatives with a free carboxyl group in their side chains were synthesized. The in vivo anti-inflammatory potential of dexketoprofen derivatives was evaluated using a carrageenan-induced paw edema model of acute inflammation. Additionally, the local and systemic redox status in rats following acute administration of the compounds was assessed by measuring levels of pro-oxidative markers and the activity of antioxidant enzymes. Among the analyzed molecules, derivatives <b>2</b> and <b>4</b> exhibited the most potent in vivo anti-inflammatory activity, showing effects comparable to those of the parent compound dexketoprofen. In vitro results revealed that all newly synthesized compounds exhibited low inhibitory activity toward COX-1, whereas only compound <b>4</b> showed significant COX-2 inhibition. The stronger binding affinity of derivative <b>4</b> for COX-2 in comparison to other tested compounds is likely attributed to its ability to form multiple electrostatic interactions within the enzyme's active site. Furthermore, compounds <b>2</b> and <b>5</b> demonstrated efficacy comparable to the parent drug in restoring redox balance, indicating their potential antioxidant properties under acute inflammatory conditions. The findings of this study underscore the therapeutic potential of the novel dexketoprofen amide derivatives as dual-function agents with the capacity to modulate both inflammatory responses and oxidative stress.

Gamma delta (γδ) T cells hold great promise in adoptive cell therapy, but suffer from low tumor-targeting efficiency. Herein, we report the development of antibody-γδ T cell conjugates for enhanced tumor therapy. By evaluating different biomolecules residing on the cell surface, sialic acids-the terminal sugars of various cell-surface glycans-are identified as the optimum site for anchoring antibodies onto γδ T cells via metabolic glycan labeling with unnatural sugars containing a bioorthogonal functional group. A programmed death-ligand 1 (PD-L1)-specific nanobody (αPD-L1) is conjugated onto γδ T cells via click chemistry and the resulting αPD-L1-γδ T cells exhibit enhanced cytotoxicity towards PD-L1-positive cancer cell lines, patient-derived primary cancer cells, and xenografted tumors in living mice. Mechanistically, αPD-L1-γδ T cells target cancer cells and tumors via binding to PD-L1 and induce cancer cell pyroptosis. Furthermore, αPD-L1-γδ T cells remodel the tumor microenvironment to be immune-active, at least partially through the recruitment and activation of CD8⁺ T cells via the CCR5/CCL5 axis. This work provides a versatile strategy for chemical engineering of γδ T cells for improved therapeutic applications.

In many model organisms, the circadian system has been proposed to comprise multiple oscillators that interact to promote accuracy of the clock as well as intricacies of rhythmic outputs. In Neurospora crassa, the circadian transcriptional/translational loop comprising of the FRQ (Frequency) and WCC (White Collar Complex) proteins has been instrumental in explaining many attributes of the clock including entrainment and rhythms in development and gene expression; in addition, some non-circadian oscillations can be unmasked when the FRQ-WCC feedback loop is eliminated. These rhythms have often lost defining circadian characteristics and are potentially controlled by other oscillators, termed FRQ-less oscillators (FLOs) in Neurospora. Understanding the biology of these oscillators and their hierarchical relationship with the FRQ-WCC oscillator (FWO) are salient questions in rhythms research. In this study, we examined candidate FLO effector pathways involving peroxiredoxins (Prxs) and mTOR. We find that independent gene knockouts compromising each pathway do not alter circadian period length or decrease the amplitude of the core circadian FWO rhythm in any meaningful way in Neurospora. Our findings suggest that molecular rhythms in Prx oxidation and in mTOR activity on the chol-1 FLO oscillator are neither required for nor strongly regulate FWO components during a normal circadian day.

Through millennia of evolution, some organisms can change colors rapidly for camouflage, communication, attraction, and warning to adapt to the dynamic environment. By mimicking the dynamic coloration strategies of natural organisms, artificial materials with responsive structural colors have emerged as promising candidates due to their environmental friendliness, real-time responsiveness to stimuli, and wide color gamut. These materials have attracted immense interest in anti-counterfeiting, displays, sensing, and smart actuators. This paper aims to present a survey on the response mechanisms, color modulation strategies, and applications of the structural color materials that are developed from different principles, such as photonic crystals (PCs), liquid crystals (LCs), and metasurfaces. We first discussed various structural color materials and their colorization mechanisms. We then classified these materials according to their color regulation strategies, such as responsiveness to physical, chemical, and biological stimuli. Subsequently, we provided a comprehensive overview of their applications for anti-counterfeiting, display technologies, sensing devices and detection platforms, and the biomedical field. Finally, we discuss the future prospects of responsive structural color materials and the issues that need to be addressed for their improvement. This review not only summarizes modulation strategies for dynamic structural colors but also provides valuable references and insights for future research in the field of structural coloration.

Identifying biomarkers predicting therapy outcomes before treatment holds great promise for advancing precision medicine. Genetic variants such as the serotonin transporter gene linked polymorphic region ( 5-HTT LPR) may be associated with response to cognitive behavioral therapy in anxiety disorders, albeit results so far are controversial. Contributing to the ongoing debate, we investigated whether treatment response to a highly standardized one-session virtual reality exposure therapy (VRET) was predicted by 5-HTT LPR genotype. N=194 patients with arachnophobia (spider phobia) were genotyped for 5-HTT LPR and the functionally related single nucleotide polymorphism rs25531 and grouped into high-(L A /L A ), and low-expression (S/S, S/L G, L G /L G , S/L A, L G /L A ) genotype. At baseline, after VRET, and at a 6-month follow-up, participants underwent a standardized behavioral avoidance task (BAT) and the spider phobia questionnaire (SPQ) to assess symptom severity. Chi-square tests revealed a significant association between 5-HTT LPR/rs25531 and behavioral treatment outcome, that remained significant at the 6-month follow-up. No association was found between genotype and self-reported symptom severity measured with the SPQ. Our results support the idea that while L A /L A genotype carriers might benefit from highly standardized treatment, lower 5-HTT expression may convey risk to poorer treatment response, likely necessitating more tailored psychotherapeutic interventions to promote sufficient response.

<title>Abstract</title> <p> Background Autism Spectrum Disorder (ASD) is a lifelong neurodevelopmental condition marked by impairments in social communication, language and behavior. Both genetic and environmental factors, including prenatal exposure to valproic acid (VPA), contribute to its pathogenesis. VPA exposure during critical periods of neurodevelopment induces oxidative stress, inflammation, and serotonergic dysregulation. Kolaviron (KV), a polyphenolic extract from <italic>Garcinia kola</italic> , exhibits potent antioxidant and anti-inflammatory properties, potentially offering Neuroprotection in ASD models <italic>. The aim of our study was to evaluate whether Kolaviron could improve the VPA-induced autism model in the areas of</italic> Mitochondrial dysregulation <italic>oxidative stress, inflammation, and behavior, and to compare its effects with oxytocin on the serotonergic system.</italic> Methods Pregnant Wistar rats received a single intraperitoneal dose of VPA (600 mg/kg) on gestational day 12.5 to induce autism-like features in offspring. Male pups were weaned on postnatal day (PND) 21 and randomly assigned to receive KV (50 or 100 mg/kg, oral), oxytocin (12 µg/kg, intranasal), or saline until PND 49. Behavioral tests were conducted on PNDs 42–49. Brain tissues were collected for ELISA analysis of hippocampal 5-HTT, 5-HTR7, TNF Alfa and IL-6 levels, Immunohistochemical staining for parvalbumin was performed to assess interneuron integrity. Results KV-treated VPA-exposed rats showed significant improvements in social interaction, reduced repetitive behavior, and attenuated. Biochemically, KV decreased IL-6 levels and modulated serotonergic markers (5-HTT, 5-HTR7). Histologically, KV preserved hippocampal architecture and parvalbumin-positive interneurons, suggesting Neuroprotection. Conclusions These effects were dose-dependent, indicating KV's potential as a complementary therapeutic agent in ASD. Further studies are warranted to clarify its mechanisms and clinical relevance. </p>

<h4>Aim</h4>This study aimed to investigate the role of mean corpuscular volume (MCV) as a predictor of mortality due to COVID-19.<h4>Materials and methods</h4>This retrospective, single-center, and longitudinal study included 122 patients with COVID-19.<h4>Results</h4>Compared to the survivor's group, the non-survivors had higher MCV (92.13 ± 3.67 fL), neutrophil-to-lymphocyte ratio [NLR] (16.99 [21.31]), platelet-to-lymphocyte ratio [PLR] (350.33 [304.68]), and systemic immune-inflammation index [SII] (3684.92 [4073.25]) levels (<i>p</i> < 0.05 for all). The optimal cutoff values for predicting in-hospital COVID-19 mortality, determined by the Youden index, indicated that patients with MCV > 89 fL, NLR > 8.69, PLR > 418.08, or SII > 2149.36 were at a higher risk of death due to SARS-CoV-2 infection. The area under the curves (AUC) of NLR, SII, MCV, and PLR was sufficient for accurate prediction. COVID-19 patients with MCV > 89 fL and PLR > 418.08 were 3.65 (95% CI 1.03-12.87; <i>p</i> = 0.043) and 5.08 (95% CI 1.06-24.22; <i>p</i> = 0.041) times more likely to die than those without these values, respectively. MCV was positively correlated with age, mean corpuscular hemoglobin (MCH), urea, blood urea nitrogen (BUN), and creatinine.<h4>Conclusion</h4>MCV > 89 fL and PLR > 418.08 at the time of hospital admission were associated with an increased COVID-19 mortality risk.

Neurodegenerative disorders (NDDs), characterized by gradual decline of neuronal function and structure, present a major threat to global public health. Recent advances in neuropharmacology have opened promising avenues for novel therapeutic approaches. This review highlights promising neuropharmacological targets that may alleviate the debilitating effects of neurodegenerative disorders. This review examines established yet emerging molecular targets in neurodegeneration, including protein aggregation, synaptic dysfunction, oxidative stress, neuroinflammation, and Rho-associated protein kinase (ROCK) signaling. The review also explores ground-breaking therapeutic strategies that have transformed modern neuropharmacology. Recent advances in nanotechnology, gene therapy, immunotherapy, and <i>in silico</i> studies have revolutionized neurotherapeutics by enabling precise drug delivery, enhancing treatment efficacy, and facilitating personalized therapies. These innovations have also accelerated the discovery of novel compounds and improved prediction of therapeutic outcomes.

FOXP1 syndrome caused by FOXP1 haploinsufficiency is characterized by intellectual disability, speech, and language impairment, autistic features, and neuropsychiatric abnormalities such as anxiety and hyperactivity. Behavioral changes in patients are mirrored in Foxp1^± mice. It is shown that decreased Foxp1 in the Foxp1^± striatum results in a significant decrease in phosphodiesterase 10a (Pde10a). Predominantly expressed in medium spiny neurons (MSNs), Pde10a modulates basal ganglia circuitry. Furthermore, the Foxp1^± striatum exhibits microglial activation, reduced synaptic pruning, and dysregulation of 111 inflammatory genes. These include the downregulated P2ry12 and Fcrls, markers of homeostatic microglia, and upregulated Cd74, a marker of reactive microglia, suggesting that neuroinflammation contributes to the observed deficits. Interestingly, treatment of Foxp1^± mice with the PDE10A antagonist MP-10 (PF-2545920) immediately after birth not only corrects behavioral abnormalities, including decreased ultrasonic vocalization, hyperactivity, and anxiety but also normalizes changes in microglia morphology and synaptic pruning. Transcriptomic analysis with a neuroinflammation-specific gene panel reveals nominal gene expression changes after MP-10 treatment, including Bdnf upregulation and enrichment of neurotrophin signaling. Since FOXP1 and its signaling pathway are highly conserved, administration of MP-10 or other Pde10a antagonists may also alleviate the neurological dysfunction seen in humans with FOXP1 syndrome.

<h4>Background</h4>An immunosuppressive tumor microenvironment limits the efficacy of immunotherapy, thus patients with MSS and pMMR mCRC often face great challenges.<h4>Methods</h4>In this phase II trial, patients received Gamma Knife SBRT combined with Tislelizumab. Biomarker analysis was performed pre- and post-treatment.<h4>Results</h4>From November 2022 to July 2024, 1of 20 patients achieved CR, 13 of 20 patients achieved PR, 6 achieved SD. mPFS was 10.7 months (95% CI, 6.4-15.0). With no grade 4 events noted, common adverse events included nausea (65%), anemia (55%), and fatigue (45%). RNA sequencing indicated enhanced immune infiltration in PR patients. For patients with pMMR/MSS/MSI-L mCRC who had not responded to first and second-line therapies, the combo of Gamma Knife SBRT and tislelizumab showed high efficacy and reasonable safety. Significant post-radiotherapy improvements in the tumor's immunosuppressive microenvironment, including lower fibrosis, normalizing of tumor vasculature, and activation of the PD-1/PD-L1 checkpoint pathway were revealed by biomarker analysis.<h4>Conclusions</h4>These results imply that patients with pMMR/MSS/MSI-L mCRC who were unresponsive to the first and second-line chemotherapy, Gamma Knife SBRT with tislelizumab provides a safe and powerful later-line treatment alternative.<h4>Funding</h4>This research was supported by the Clinical Frontier Technology Program of the First Affiliated Hospital of Jinan University (No. JNU1AF-CFTP-2022-a01223), the National Natural Science Foundation of China (82204436), Natural Science Foundation of Guangdong Province (2024A1515030010, 2022A1515011695), Science and Technology Projects in Guangzhou (2024A03J0825).<h4>Clinical trial number</h4>ChiCTR2200066117.

Understanding the mechanisms underlying Kirsten rat sarcoma (KRAS) mutation-driven development and progression of pancreatic ductal adenocarcinoma (PDAC) may facilitate the discovery of novel strategies for KRAS-mutant PDAC (KRAS^mut-PDAC) treatment. Here, it is reported that downregulation of arachidonate 15-lipoxygenase (ALOX15B) significantly correlated with poor outcomes in patients with KRAS^mut-PDAC. Mechanistically, KRAS^mut/ERK1-elicited phosphorylation of ABHD17C promotes depalmitoylation and membrane-to-cytoplasm translocation of ALOX15B, facilitating proteasome-dependent degradation of ALOX15B via interaction with the E3 ligase complex CUL4/DDB1/DCAF10. Notably, treatment with methyl protodioscin (MPD), a steroid saponin primarily purified from polygonatum sibiricum rhizome, restored the S-palmitoylation and membrane location of ALOX15B via disruption of the ABHD17C/ALOX15B interaction, consequently resulting in significant inhibition of growth rate of patient-derived KRAS^mut-PDAC organoids in vitro and KRAS^mut-PDAC-formed tumor in vivo via induction of ferroptosis. Therefore, these findings unveil a prominent role of ferroptosis evasion in KRAS^mut-PDAC progression and highlight the potential of targeting KRAS/ERK1/ABHD17C/ALOX15B axis in KRAS^mut-PDAC treatment.

Despite the growing number of older and aging people in custody across many countries, and corresponding vulnerability for neurocognitive issues, there has been limited research into the efficacy of cognitive assessment and dementia screening tools in correctional settings. The purpose of this study was to synthesize recent empirical research investigating dementia (and cognitive impairment) in older people in custody using screening tools. A systematic review methodology was adopted, with reporting guided by the PRISMA framework. This included a systematic search of four databases (PsycINFO, MedLine, PubMed, and Academic Search Complete) and handsearching of reference lists of eligible studies. A total of 9 peer-reviewed publications were included. All studies were from high income countries, including the United States (<i>n</i> = 2), Germany (<i>n</i> = 2), Canada (<i>n</i> = 1), England and Wales (<i>n</i> = 2), Australia (<i>n</i> = 1), and France (<i>n</i> = 1). These studies were synthesized according to the following themes: (1) use of screening tools with modification for the correctional setting; (2) use of screening tools without modification for the correctional setting; and, (3) innovative advances in cognitive screening tools and practices for correctional settings. This review identified several cognitive assessment and dementia screening tools that have been used in custodial settings. These tools can help in the early identification of dementia (or cognitive impairment) among older people in custody and, thus, inform the initiation of supports and strategies to manage those at risk. More research is needed to evaluate the performance of these tools compared with a diagnostic assessment.

Lysosomes are cytoplasmic organelles central for the degradation of macromolecules to maintain cellular homoeostasis and health. However, how lysosomal activity can be boosted to counteract ageing and ageing-related diseases remains elusive. Here we reveal that silencing specific vacuolar H⁺-ATPase subunits (for example, vha-6), which are essential for intestinal lumen acidification in Caenorhabditis elegans, extends lifespan by ~60%. This longevity phenotype can be explained by an adaptive transcriptional response typified by induction of a set of transcripts involved in lysosomal function and proteolysis, which we termed the lysosomal surveillance response (LySR). LySR activation is characterized by boosted lysosomal activity and enhanced clearance of protein aggregates in worm models of Alzheimer's disease, Huntington's disease and amyotrophic lateral sclerosis, thereby improving fitness. The GATA transcription factor ELT-2 governs the LySR programme and its associated beneficial effects. Activating the LySR pathway may therefore represent an attractive mechanism to reduce proteotoxicity and, as such, potentially extend healthspan.

The special AT-rich sequence binding protein 1 (SATB1) has been linked to neurodevelopmental disorders (NDDs) including developmental delay, intellectual disabilities (ID) and autism spectrum disorder (ASD). But the underlying biological mechanisms are still not fully understood. In this study we generated a rat model with a truncated Satb1 protein. We showed that Satb1 mutant caused growth retardation, microcephaly, altered ultrasonic vocalization and delayed neurobehavioral development in mutant pups as well as social and cognitive behavior deficits in adult mutants, mimicking the typical clinical characteristics of SATB1-associated NDDs. Injection of a GABAergic enhancer clonazepam (0.04 mg/kg, i.p.) effectively alleviated the abnormal social and cognitive behaviors in Satb1 mutant rats. Finally, RNA sequencing analysis further revealed a potential role of Satb1 in a cortical transcriptional regulatory network associated with NDDs including ID and ASD. Our results confirm the crucial roles of SATB1 in the pathogenesis of NDDs and provide insights into treatment strategies for SATB1-associated NDDs.

Intracranial atherosclerosis (ICAS), a common cause of ischemic stroke, remains a therapeutic challenge due to complex intracranial anatomy and intervention risks. Although ICAS develops intracranially, cerebral artery innervation originates in the peripheral nervous system. The scarcity of human intracranial specimens has hindered investigations into the potential role of Schwann cells (SCs) in neurovascular homeostasis. Using multimodal analysis of plaque-bearing and non-plaque-bearing intracranial artery samples from the same postmortem individuals (n = 16 donors), SCs associated with myelinated neural structures are identified. Quantitative ultrastructural evaluation reveals a 4.3-fold increase in SC-derived myelin sheaths within plaque-bearing vessels (P <0.001). Single-cell RNA sequencing (scRNA-seq) of SCs demonstrates significant upregulation of genes involved in axonogenesis, axon ensheathment, axon guidance, synaptic transmission, and synaptic integration. Cell-cell communication analysis shows enhanced interactions between SCs and vascular smooth muscle cells (VSMCs) in plaque-bearing vessels. Synaptic-like structures are observed in the walls of intracranial arteries, along with a 2.9-fold increase in VSMC-innervating myelinated fibers (P <0.001). Ligand-receptor analysis indicates SPP1-ITGB1 signaling as a potential mediator of SCs-VSMCs crosstalk. This study provides evidence for the involvement of SCs in ICAS pathobiology and proposes novel neurovascular targets for precision therapies in cerebrovascular disease.

<h4>Introduction</h4>Type 3 hereditary hemochromatosis (HH) is a rare genetic disease due to mutations in the transferrin receptor 2 (<i>TFR2</i>) gene.<h4>Methods and results</h4>Here, we describe the case of an Italian patient presenting with hyperferritinemia and hepatic iron accumulation, not evidenced by magnetic resonance imaging, that was subsequently classified as HH Type 3 by the identification of the novel frameshift mutation c.523_524delC>T (p. Leu175Aspfs*41) in exon 4 of <i>TFR2</i> gene through the whole exome sequencing (WES) approach.<h4>Conclusion</h4>WES would allow to diagnose rare HH-related diseases in patients with unexplained hepatic iron overload and/or aberrant circulating iron parameters.<h4>Trial registration</h4>The authors have confirmed clinical trial registration is not needed for this submission.

Microgravity accelerates skeletal muscle degeneration, mimicking aspects of aging, yet its effects on muscle cell function remain underexplored. Using a muscle lab-on-chip model onboard the International Space Station (ISS), we examined 3D-bioengineered myobundles derived from young and older adult donors under microgravity. Electrical stimulation applied intermittently to the myobundles revealed reduced contraction magnitude in microgravity and decreased protein levels of myosin heavy chain 7, a main isoform in slow-twitch muscle fibers. Transcriptomic profiling revealed active myogenesis across ground and spaceflight samples, but younger electrically stimulated myobundles displayed enhanced mitochondrial-related gene expression in microgravity, while older and non-electrically stimulated myobundles were less responsive. Comparative analysis between young and older derived myobundles identified 86 muscle-specific age-associated genes altered in microgravity, linked to inflammation, mitochondrial dysfunction, and cellular stress. These findings highlight a unique age-related molecular response in microgravity and underscores electrical stimulation as a potential countermeasure. These insights advance our understanding of muscle aging and degeneration in microgravity, guiding future therapeutic strategies.

Epithelial serous borderline tumors (SBT) are non-invasive neoplastic ovarian lesions that may recur as chemo-resistant low-grade serous cancer (LGSC). While genetic alterations suggest a common origin, the transition from SBT to LGSC remains poorly understood. Here, we integrate cell-type resolved spatial proteomics and transcriptomics to elucidate the evolution from SBT to LGSC and its corresponding metastases in both stroma and tumor. The transition occurs within the epithelial compartment through an intermediary stage with micropapillary features, during which LGSC overexpresses c-Met and several brain-specific proteins. Within the tumor microenvironment, interconnectivity between cancer and stromal cells, along with enzymes degrading a packed extracellular matrix, suggests functional collaboration among various cell types. We functionally validated 16 drug targets identified through integrated spatial transcriptomics and proteomics. Combined treatment targeting CDK4/6 (milciclib) and FOLR1 (mirvetuximab) achieved significant tumor reduction in vivo, representing a promising therapeutic strategy for LGSC.

Axon guidance signaling pathways, including the Eph/ephrin, Semaphorin, and Slit/Robo pathways, have been found to play crucial roles in cardiac development. Netrin signaling is another well-studied signaling pathway important for axon guidance, but its role in the developing heart has not been investigated. Here, we describe the novel expression pattern of Netrin-1 in the developing murine heart. Transcriptomic analysis of embryonic mouse hearts shows dynamic Netrin-1 expression from E8.5 through E14.5, where Netrin-1 expression preferentially co-localizes with developing trabecular cardiomyocytes. We further demonstrate the spatiotemporal expression pattern of Netrin-1 using a combination of RNA in situ hybridization and Netrin-1^Bgeo/+ reporter mice. Netrin-1 is expressed in the developing cardiomyocytes with the highest degree of expression within the left ventricular trabecular myocardium, which has not been previously recognized. Additionally, Netrin-1 expression is observed at lower levels in the cardiomyocytes of the right ventricle and atria. This expression pattern supports a role for Netrin signaling in the developing murine myocardium requiring further functional characterization.

Cellular stress causes DNA strand breaks that are typically repaired to maintain homeostasis and regulate cell fate. However, unrepaired DNA breaks can be lethal, leading to cell death. Here, we present a protocol to study DNA strand breaks in Drosophila during development and apoptosis using in situ nick translation. We describe the steps for labeling DNA strand breaks using digoxigenin (DIG)-labeled nucleotide (DIG-11-dUTP) and visualizing them with anti-DIG immunostaining. We then detail procedures for mounting, imaging, and analysis. For complete details on the use and execution of this protocol, please refer to Maurya et al.¹ and Rigby et al.².

Autism spectrum disorder (ASD) is a neurodevelopmental condition with a complex genetic basis. Large-scale whole-exome sequencing and whole-genome sequencing studies, with increasing sample sizes and improved ancestral diversity, have significantly advanced the discovery of ASD-associated genes. In addition to identifying coding variants, whole-genome sequencing has facilitated the detection of risk noncoding variants in regulatory elements such as enhancers, promoters, and untranslated regions, prompting experimental validation of their functional impact on neurodevelopment. A deeper understanding of ASD genetic liability has revealed the interplay between rare and common variants. Moreover, genetic liability varies by sex and phenotype profile, underscoring the complexity of ASD's genetic architecture. While the clinical application of these genomic insights remains in early stages, progress has been made in gene-based therapeutic development, the interpretation of noncoding risk variants, and the use of polygenic score for risk stratification. In this review, we summarize key findings from large-scale genomic studies, explore the role of coding and noncoding variants in ASD, and discuss emerging opportunities for translating these discoveries into clinical practice.

<h4>Background</h4>The diagnosis of genetic hyperkinetic movement disorders has become increasingly more complex as new genes are discovered and technologies offer new diagnostic possibilities. As a result, the choice of appropriate gene testing and the interpretation of the results can become difficult to navigate for movement disorder experts and clinicians. In parallel, research is becoming crucial to pair with clinical assessments in order to explore advanced sequencing technologies and allow new genes discovery.<h4>Methods</h4>Systematic review of genetic forms of hyperkinetic movement disorders and of the most relevant genetic terminology was performed.<h4>Results</h4>Comprehensive descriptions of genetic lexicon, testing selection, and complex genetic findings related to hyperkinetic movement disorders are reported.<h4>Discussion</h4>Here we discuss the terminology of genetic diagnosis that is now part of the clinical practice, the difficulties related to the interpretation of complex genetic results, and provide guidance and tips for gene testing selection in order not to miss important diagnosis of genetic hyperkinetic movement disorders.<h4>Highlights</h4>To review the most relevant lexicon related to genetic diagnosis, approach to gene testing, testing selection, and complex genetic findings in genetic hyperkinetic movement disorders.

Circular RNA (circRNA) is a prominent group of non-coding RNA (ncRNA) that regulates gene expression at the post-transcriptional level in various diseases including acute respiratory distress syndrome (ARDS). In our study, circVAMP3 was a novel circRNA significantly increased in macrophages in an acute lung injury (ALI) model. Through knockdown and overexpression of circVAMP3, we found circVAMP3 promoted the expression of IFN-β in macrophages, which was vital in the pathophysiological process of ARDS. Moreover, we showed that circVAMP3 targeted Stimulator of Interferon Genes (STING) through miR-580-3p by bioinformatics analysis and intervene of miR-580-3p. CircVAMP3 acts as a pro-inflammation ncRNA by regulating the miR-580-3p/STING/IFN-β axis and could be a potential biomarker and therapeutic target for ARDS.

Depression is the most common psychiatric comorbidity of epilepsy. While antiseizure medications (ASMs) can exacerbate depressive symptoms, depression itself may increase both the frequency and duration of epileptic seizures. The pathophysiological mechanisms underlying epilepsy-depression comorbidity (EDC) involve neurotransmitter imbalance, inflammatory responses, oxidative stress (OS), gut microbiota dysbiosis, and neuroendocrine abnormalities. Recent studies highlight that inflammation contributes to EDC via multiple interconnected mechanisms, including glial cell activation, cytokine release, pyroptosis, and oxidative damage, ultimately leading to synaptic dysfunction and neuroimmune imbalance. Drawing from representative and recent evidence, this review summarizes the role of inflammation in the pathogenesis and progression of EDC. It also outlines current inflammation-targeted therapeutic strategies, such as anti-inflammatory drugs, antioxidants, herbal medicine, acupuncture, probiotic modulation, and precision therapies. This review provides a conceptual framework for understanding inflammation-mediated EDC and offers insights into targeted treatment approaches.

DNA methylation is the best-known epigenetic mechanism regulating gene expression without altering the DNA sequence. Its counterpart, known as DNA demethylation, is equally important and enables the activation of previously silenced genes. DNA demethylation has attracted interest in the scientific community following the landmark discovery that Ten-Eleven Translocation (TET) proteins can convert 5-methylcytosine to 5-hydroxymethylcytosine. A growing body of research indicates that changes in TET protein levels and 5-hydroxymethylcytosine content are hallmarks of cancer. These epigenetic changes appear to play a critical role in the development of malignancies characterized by high levels of somatic mutations and genetic instability. Bladder cancer is among the most common cancers worldwide and, despite aggressive treatment, remains associated with high mortality and poor prognosis. The lack of reliable diagnostic and prognostic markers poses a significant challenge in its management, highlighting the urgent need for novel biomarkers to enable earlier diagnosis and more accurate prediction of clinical outcomes. This review examines epigenetic alterations associated with bladder cancer and their clinical implications. We focus on the impact of DNA methylation and demethylation on oncogene regulation, summarize scientific evidence supporting their role in bladder cancer development and progression, and briefly explore novel therapeutic strategies targeting those epigenetic mechanisms.

Physical strength and endurance of an individual are vital for athletic performance, and minimizing the risk of injuries, especially during competitions. Other than training and diet, athletic performance is determined by genetic factors or heredity, which is less focused in sports science research. Genetic factors play a crucial role in greater cardiovascular endurance and muscular phenotypes, and thereby contribute to athletic success. Several genes and different polymorphisms are positively/negatively associated with athletic performance. This review delved into the intricate role of several genes and polymorphisms in different-population groups, and explored their impact on an individual's ability to engage in athletic activities. Among several identified genes, the prominent genes, including <i>ACE</i>, <i>AGT</i>, <i>BDKRB2</i>, <i>NOS3</i>, <i>HIF1A</i>, <i>ACTN3</i>, <i>AMPD1</i>, <i>PPARGC1</i>, <i>SOD2</i>, <i>BDNF</i>, <i>VDR</i> and mtDNA are discussed in this study. These genes have been reported to play indispensable roles in endurance performance and power. Furthermore, genetic variations/polymorphisms within these genes are potential to impact various aspects of physiology, including cardiovascular function, muscle fiber composition, and metabolic efficiency. Genetic polymorphisms are recognized as contributing factors in determining the athletic capacity to engage and perform sustained physical activities in their respective sports. We emphasized the noteworthy discoveries from the existing literature, and precisely explored the association between particular gene polymorphisms and athletic prowess, with a specific focus on endurance-oriented sports (running, cycling, and swimming) and power sports. Understanding the genetic variations and their influence on endurance/power sports can offer valuable insights for athletes, coaches, and scientists in sports sciences, who strive to enhance athletic training strategies and performance outcomes in achieving success.

Glioblastoma is a highly aggressive, infiltrative brain tumor of the central nervous system (CNS). Its extensive molecular and biochemical heterogenicity hinders the identification of reliable biomarkers and therapeutic targets, thereby making prognosis and existing therapy ineffective. In recent years, breakthroughs in the use of proteomics on a range of biological samples, such as plasma, cerebrospinal fluid (CSF), tissues, brain cells, and exosomes, represent a potential improvement to GBM investigations. Mass spectrometry-based approaches represent an important technique in the characterization of the tumoral proteome, for the identification of differentially expressed proteins, and for studying altered molecular pathways involved in tumor stages. Proteomics studies advance our knowledge about GBM pathogenesis, the discovery of reliable diagnostic and prognostic markers, and therapeutic approaches, also. In this context, for the effective application of proteomics on GBM, it is mandatory to develop a translational network by integrating hospitals, biobanks, and research institutions into a single network, to enable a collaborative approach across disciplines, thereby enabling rapid translation to clinical application of new proteomic insights. Today, high-quality biobanks play a key role in enabling collaborative, ethically compliant research, supporting the effective application of proteomics in glioblastoma studies and the translation of discoveries into clinical practice. This review explores current trends in proteomics and GBM research, highlighting how leveraging biobank infrastructure and fostering institutional cooperation can drive the development of targeted pilot projects to enhance the impact and effectiveness of glioblastoma research.

<i>Background/Objectives:</i> Colorectal cancer (CRC) remains a major global health burden, marked by complex tumor-microenvironment interactions, genetic heterogeneity, and varied treatment responses. Effective preclinical models are essential for dissecting CRC biology and guiding personalized therapeutic strategies. This review aims to critically evaluate current experimental CRC models, assessing their translational relevance, limitations, and potential for integration into precision oncology. <i>Methods:</i> A systematic literature search was conducted across PubMed, Scopus, and Web of Science, focusing on studies employing defined in vitro, in vivo, and emerging integrative CRC models. Studies were included based on experimental rigor and relevance to therapeutic or mechanistic investigation. Models were compared based on molecular fidelity, tumorigenic capacity, immune interactions, and predictive utility. <i>Results:</i> CRC models were classified into in vitro (2D cell lines, spheroids, patient-derived organoids), in vivo (murine, zebrafish, porcine, canine), and integrative platforms (tumor-on-chip systems, humanized mice, AI-augmented simulations). Traditional models offer accessibility and mechanistic insight, while advanced systems better mimic human tumor complexity, immune landscapes, and treatment response. Tumor-on-chip and AI-driven models show promise in simulating dynamic tumor behavior and predicting clinical outcomes. Cross-platform integration enhances translational validity and enables iterative model refinement. <i>Conclusions:</i> Strategic deployment of complementary CRC models is critical for advancing translational research. This review provides a roadmap for aligning model capabilities with specific research goals, advocating for integrated, patient-relevant systems to improve therapeutic development. Enhancing model fidelity and interoperability is key to accelerating the bench-to-bedside translation in colorectal cancer care.

Pollen, the male gametophyte of flowering plants, is collected by honeybees as a primary source of protein and converted into bee pollen through the enzymatic activity of digestive secretions. The nutrients in bee pollen are available in amounts well beyond those of proteins, comprising macronutrients such as carbohydrates, lipids and dietary fiber, as well as micronutrients such as minerals, vitamins, organic acids, and phenolic compounds. This study aimed to determine the macro and trace mineral content of bee pollen from different botanical and geographical origins, and to assess their bioaccessibility through simulated in vitro digestion, their dietary contribution, and potential health risks. Seven bee pollen samples were investigated, three with a monofloral origin of above 80%, from <i>Nigella</i> spp., <i>Helianthus annuus</i> and <i>Castanea sativa</i>, and four with a multifloral origin. Mineral composition revealed potassium as the most abundant element, while iron, manganese, and copper were found at trace levels. <i>Castanea sativa</i> pollen had the highest overall mineral content, whereas <i>Nigella</i> spp. showed the lowest values for calcium, magnesium, and copper. The bioaccessibility of bee pollen was highest during the gastric phase for most minerals except copper, where most of the samples peaked in the intestinal phase. Overall, mineral bioaccessibility after simulated digestion followed the order K > Mg > Cu > Ca > Mn > Fe > Zn. While for manganese, the consumption of bee pollen showed the highest contribution to recommended dietary intake (16% for women and 12% for men), calcium had the lowest, with less than 1% of the RDA at a consumption level of 40 g/day. Health risk assessment confirmed that consuming 40 g/day of bee pollen poses no risk because the target hazard quotient and hazard index are below the risk threshold of 1.0.

The family of voltage-dependent anion channels (VDACs) comprises three isoforms (VDAC-1, VDAC-2, VDAC-3). VDACs have been extensively described as localised in the outer mitochondrial membrane where they are involved in the exchange of ions, metabolites, and ATP/ADP between mitochondria and cytosol. The VDAC interacts with disease-specific proteins and thus regulates the mitochondrial function and controls the cellular energy resources, explaining its involvement in cell death and apoptosis. In addition, VDAC-1 and -2 can also be found at other cellular locations such as in the sarcoplasmic reticulum, in the endoplasmic reticulum, as well as in the plasma membrane. Through single-channel pore regulation, oligomerisation, or changed expression levels the VDAC is involved in different neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Huntington's disease, and others. Here, we critically summarise current discussions about the VDAC as a common key player for these diseases. We suggest that the VDAC acts as a transmembrane multifunctional regulatory protein which might serve as a pharmacological target for the development of novel drugs against neurodegenerative diseases such as the application of recombinant antibody technology.

Metabolic dysfunction-associated steatotic liver disease (MASLD) has been consistently linked to increased risk of cardiovascular disease (CVD). HDL lipoproteins may serve as a possible link in this association through their hepatic synthesis and atheroprotective properties. Serum samples were collected from 51 MASLD patients (diagnosed by abdominal ultrasound), 40 with coronary artery disease, and 50 healthy controls. HDL lipid profiles were investigated by proton nuclear magnetic resonance (¹H NMR) spectroscopy. Patients with MASLD exhibit an increased percentage of lysophosphatidylcholine and sphingolipid content, mainly due to increased ceramides, and a reduced percentage of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol compared to controls. The % content of total and individual polyunsaturated fatty acids including linoleic, docosahexaenoic, eicosapentaenoic, and arachidonic acid was found to be reduced in patients with MASLD, while saturated fatty acid content was increased compared to the control group. These alterations in fatty acid composition were observed also in CAD patients compared to controls but were more pronounced in CAD patients. Compared to CAD patients, those with MASLD showed an increased content of sphingolipids, ceramides, and glycerolipids and a reduced content of phosphatidylinositol. Changes observed in the lipid composition of HDL lipoproteins in MASLD patients may impair the protective properties of HDL particles, contributing to increased CVD risk.

The gut microbiota constitutes a complex community of microorganisms (including bacteria, viruses, fungi, and protozoa) within the intestinal tract. Over the years, an increasing number of studies have highlighted the bidirectional communication between the gut microbiota and the central nervous system (CNS), a relationship commonly referred to as the "microbiota-gut-brain axis". In particular, the crosstalk between the gut microbiota and the brain has been associated with the pathogenesis and progression of various CNS disorders. Phages, or bacteriophages, viruses that specifically infect bacteria, constitute the most abundant viral component within the gut microbiota. However, despite their abundance and significance in the gut microbial community, studies exploring the relationship between phages and the CNS remain surprisingly limited. This review examines the biological interplay between gut-resident phages and the CNS. Furthermore, we discuss the current literature linking phages to CNS-related pathologies.

Recent advancements in cell and tissue biology have fundamentally changed our understanding of cellular behavior, revealing that both stem and nonstem cells exhibit remarkable plasticity and adaptability. This discovery has paved the way for revolutionary medical drug therapies that leverage cell and tissue reprogramming to repair or regenerate damaged tissues, offering new hope for conditions that were once considered irreversible. Tissue reprogramming involves the activation of specific molecular pathways to convert the function of residual tissue to compensate for the loss of tissue function to aging, trauma, or disease processes. By targeting these pathways, emerging drugs can promote regenerative processes, enabling the restoration of tissue function lost due to aging, injury, or disease. These therapies have shown promising results in preclinical studies addressing a wide range of diseases. Unlike traditional treatments, which focus primarily on managing symptoms, tissue reprogramming therapies offer a dynamic approach that can fundamentally alter cellular states, leading to functional recovery. This review explores the current state of cell and tissue reprogramming, highlighting its potential applications in regenerative medicine and the challenges that must be addressed for successful clinical translation. As our understanding of cellular plasticity continues to evolve, these innovative therapies stand at the forefront of a new era in medicine, with the potential to transform treatment paradigms and significantly improve patient outcomes across a wide range of conditions. SIGNIFICANCE STATEMENT: Breakthrough technologies have transformed our understanding of cell and tissue biology, uncovering that cells and tissues possess remarkable adaptability and fluidity in their roles. This revelation has opened up exciting possibilities in regenerative medicine, where emerging drug therapies aim to harness and reprogram cells to repair or regenerate damaged tissues. An emerging class of medical drugs will activate the body's natural regenerative abilities, offering the potential to restore tissue function lost due to aging, injury, or disease.

Pneumonia is a major issue that affects calves' health and performance and causes numerous losses despite treatment. Investigating genetic and molecular differences, as well as immunological and antioxidant responses, in calves at risk for pneumonia was the aim of this study. A total of 225 calves were studied, including 180 Holstein calves with respiratory signs and 45 calves that were apparently healthy. Blood samples were collected for CBC, RNA extractions, and immunological and antioxidant analysis. In contrast to the control group, the pneumonic one showed a considerable (<i>p</i> < 0.05) increase in the expression levels of cytokines and antioxidant genes IL1α, IL-1β, IL-6, IFN-γ, TNF-α, and NOX4. In contrast, the values of IL10, PRDX6, ATG7, and NDUFS6 were in the opposite range. The pneumonic and healthy calves were found to differ in the nucleotide sequences of the genes under analysis. In pneumonic calves, a substantial (<i>p</i> ˂0.05) reduction was detected in RBCs, Hb count, PCV%, and lymphocytes count, and a notable (<i>p</i> ˂ 0.05) increase in WBCs and neutrophil count was correlated with healthy control calves. The findings of the serum profile showed that there was a meaningful (<i>p</i> ˂ 0.05) rise in the serum values of IL-1α, IL-1β, IL-6, TNF-α, IFN-γ, and MDA, with significant reductions in the SOD, GSH, TAC, and IL-10 in the pneumonic compared to the healthy calves. Our results provide valuable information about the nucleotide sequence, gene expression, and serum profile differences of putative indicators for pneumonia in calves. This could be applied in monitoring calves' pneumonia through the discriminate breeding of naturally resistant animals.

End-stage renal disease (ESRD) is a pathophysiological condition that requires radical solutions for treatment. Although ESRD is a worldwide health problem, the incidence of the disease is relatively high in the Mediterranean island of Cyprus. Moreover, the etiology of the disease largely remains poorly understood. Overlapping diseases such as diabetes mellitus may pose a challenge during the differential diagnosis of ESRD. Hence, molecular diagnosis could be powerful in terms of ESRD diagnosis. Nevertheless, the genetic isolation of Cyprus and the complexity of the disease necessitate more comprehensive, population-based approaches. In the current study, exome sequencing (ES) data obtained from 24 Turkish Cypriot patients with ESRD were re-analyzed. Variants were filtered according to the allele frequencies in population databases and the renal anomalies-related gene list. Variants were then prioritized according to the guidelines of the American College of Medical Genetics. The re-analysis underscored that different genes and their variants could explain the molecular mechanisms of the disease. However, numerous variants were found to be prevalent in all patients, limiting the ability to reach a population-based conclusion about the aetiology of ESRD in Cyprus. Although ES could be useful for the molecular diagnosis of ESRD, further population-based cohort studies and a whole genome database with data from healthy participants are still needed.

Positron emission tomography (PET) imaging targeting glypican-3 (GPC3) holds promise for improving the detection and characterization of hepatocellular carcinoma (HCC). Preclinical and early clinical studies have largely utilized high-molecular-weight antibodies radiolabeled with isotopes such as ⁸⁹Zr and ¹²⁴I, demonstrating high affinity and tumor uptake but suffering from prolonged circulation times and suboptimal signal-to-background ratios. To address these limitations, interest has shifted toward low-molecular-weight vectors-synthetic peptides and small antibody fragments-labeled with shorter-lived radionuclides (e.g., ⁶⁸Ga and ¹⁸F) to enable rapid pharmacokinetics and same-day imaging protocols. Emerging platforms such as affibodies and aptamers offer further advantages in target affinity and reduced immunogenicity. However, clinical translation requires rigorous validation: larger, histologically confirmed cohorts, head-to-head comparison with CT/MRI, and correlation with hard clinical endpoints. Moreover, leveraging GPC3 expression as a biomarker could guarantee a deeper knowledge of tumor biology-differentiation grade and vascular invasion risk-and guide theranostic strategies. While β-emitters (⁹⁰Y, ¹⁷⁷Lu) have been explored for GPC3-directed therapy, their efficacy is influenced by oxygenation and cell-cycle status, whereas α-emitters (²²⁵Ac) may overcome these constraints, albeit with challenges in radionuclide selection and daughter nuclide management. Finally, dual-targeting probes combining GPC3 and prostate-specific membrane antigen (PSMA) have demonstrated superior uptake and retention in murine models, suggesting a versatile approach for future clinical diagnostics and therapy planning.

We report on the synthesis and characterization of a novel fluorenyl-methoxycarbonyl (Fmoc)-containing thioxo-triazole-bearing dipeptide <b>5</b>, evaluated for potential therapeutic applications. The compound was tested for its antioxidant and antimicrobial properties, demonstrating significant effects in scavenging reactive oxygen species (ROS) and inhibiting microbial growth, particularly when combined with plant extracts from an endemic <i>Peonia</i> species from the Caucasus. Circular dichroism (CD) binding studies with bovine serum albumin (BSA) and calf thymus DNA revealed important interactions, suggesting the dipeptide's potential in biomedically relevant conditions that involve DNA modulation. Molecular docking and CD spectra deconvolution provided additional insights into the binding mechanisms and structural characteristics of the formed complexes with the biomolecular targets. The Fmoc group enhances the dipeptide's lipophilicity, which may facilitate its interaction with cellular membranes, supporting efficient drug delivery. A computational evaluation at the ωB97XD/aug-cc-pVDZ level of theory was carried out, confirming the experimental results and revealing a powerful potential of the peptide as an antioxidant, through FMOs, MEP analysis, and antioxidant mechanism assessments. Together, these findings suggest that this dipeptide could be valuable as an antimicrobial and antioxidant agent, with potential applications in pathologies involving oxidative stress, DNA modulation, and microbial infections.

<h4>Background/objectives</h4>Bovine respiratory disease (BRD) is a common disease impacting cattle throughout the US. BRD is a multifactorial disease as disease risk varies with the genetic profile of the host, environmental conditions, and pathogen exposure. Selection for enhanced BRD resistant cattle can aid in reducing BRD. The objectives of this study were to identify loci, gene sets, and genes associated and enriched for BRD in pre- and post-weaned Holstein cattle.<h4>Methods</h4>Cases consisted of 2147 and 5607 calves treated for BRD as pre-weaned (0-60 days old) and post-weaned (61-420 days old) calves, respectively. Controls consisted of calves untreated for BRD that remained in the herd for 61 (<i>n</i> = 14,219) days for pre-weaned or 421 (<i>n</i> = 12,242) days for post-weaned calves. A genome-wide association analysis (GWAA) identified loci and positional candidate genes associated with BRD (uncorrected <i>P</i> < 1 × 10^-5) for additive, dominant, and recessive inheritance models. A gene set enrichment analysis (GSEA-SNP) identified gene sets and leading-edge genes enriched (NES ≥ 3) for BRD.<h4>Results</h4>In pre-weaned calves, 62 loci and 123 positional candidate genes were associated (<i>P</i> < 1 × 10^-5) in addition to the 12 gene sets and 126 leading-edge genes enriched (NES ≥ 3) for BRD. In post-weaned calves, 181 loci and 185 positional candidate genes were associated (<i>P</i> < 1 × 10^-5), and 63 gene sets and 849 leading-edge genes were enriched (NES ≥ 3) for BRD.<h4>Conclusions</h4>These results provide further insight and validation of genomic regions that enhance selection for BRD resistance and for healthier cattle.

<h4>Background</h4>The heart is the first functional organ to develop in the vertebrate embryos. In mice, the primitive tubular heart begins beating at embryonic day (E) 8.0-E.8.5 and undergoes rightward looping to form the atrial and ventricular chambers. The proepicardium, a transient cell cluster at the sinus venous-lateral plate mesenchyme junction migrates onto the heart and gives rise to the embryonic epicardium, a squamous epithelium that plays a key role in cardiac development. Despite advances in understanding epicardial lineage contributions, the molecular mechanisms governing these processes remain poorly understood.<h4>Methods</h4>To characterize the transcriptional and post-transcriptional regulation of epicardial development, we performed RNA sequencing at two critical timepoints, proepicardium formation and embryonic epicardium establishment. We analysed differentially expressed coding and non-coding RNAs, focusing on microRNAs and their potential regulatory interactions.<h4>Results</h4>We identified a complex network involving differentially expressed mRNAs, microRNAs and lncRNAs between proepicardium and embryonic epicardium. Notably, with miR-495 and let-7c emerged as key regulators of epicardial cell migration, an essential process for proper epicardium formation and epicardial-derived cell migration. Our findings also reveal that these microRNAs not only regulate target gene expression but also modulate other microRNAs, suggesting a novel regulatory mechanism in epicardial development. Additionally, Foxf1 inhibition modulates let-7c, promoting the expression of key cardiogenic lineage markers in epicardial cells.<h4>Conclusion</h4>Our study highlights the role of Foxf1 in regulating miR-495 and let-7c, which in turn modulate epicardial cell migration and myocardial specification. These finding provide new insights into the intricate interplay between transcription factors and microRNAs in governing cardiogenesis.

Clinical care for patients with limited life expectancy often requires adjustments, prioritizing immediate benefits over long-term outcomes, as the relevance of future complications diminishes. This study identifies indicators of mortality risk in horses with chronic orthopaedic conditions to enhance individualized care and welfare. Over 3 years, 123 chronically lame horses and 6 healthy control horses at an animal sanctuary underwent regular (every 3 months) comprehensive health assessments and activity monitoring using wearable sensors. Data collected included body condition scores, musculoskeletal pain scores, lameness evaluations, and time budgets for eating, resting, and activity. Of the 123 chronically lame horses, 31 horses died (n = 31/123, 25.2%), with 10 succumbing to acute decompensation of their chronic condition (DAC, n = 10/123, 8.1%), while 21 were euthanized due to intractable pain or progressively deteriorating health and function (DCC, n = 21/123, 17.1%). Statistical modelling using death as outcome measure revealed body condition, pain scores, and time budget data to be strongly associated with equine mortality. Notably, low body condition score and reduced eating time predicted mortality in DAC horses, aligning with human studies linking weight loss to frailty and increased mortality risk. Additionally, depression-like behaviours were prevalent in DAC horses, mirroring the link between depression and mortality in humans. While pain scores were elevated in all deceased horses, weight loss was specific to DAC, suggesting multifactorial influences beyond pain. These findings provide a foundation for developing equine-specific tools to predict outcomes and guide clinical and end-of-life decisions, enabling individualized treatment to enhance the welfare and quality of life for aging horses. These insights may also offer valuable information for human medicine, particularly for at-risk groups such as individuals with cognitive impairments who may struggle to communicate their symptoms.

The fillet yield phenotype is a trait that can be improved in aquaculture species through conventional selective breeding. This approach was applied to rainbow trout for three consecutive generations of selection to produce a high-yield line (HY) that exhibits 2.5 percentage points higher fillet yield compared to a low-yield line (LY). To characterize the genetic and physiological mechanisms contributing to the HY phenotype, transcriptomic analysis of liver and skeletal muscle was performed at three stages of development, 2 g, 60 g, and 300 g, which corresponded to 35, 208, and 277 days post-hatch. Functional analysis of differentially expressed genes (DEG) suggests that increased muscle yield in the HY line is partially driven by greater hyperplasia at 60 g; although, higher rates of protein accretion, primarily attributed to lower rates of protein degradation, promote muscle cell hypertrophy during all stages of development. Additionally, DEGs support reductions in glycolysis in the HY muscle, with increased activity of the more efficient citric acid cycle and oxidative phosphorylation reactions for energy production compared to the LY line. In the liver, DEGs indicate unique nutrient utilization mechanisms in the HY line that support reduced visceral adiposity compared to the LY line. These findings provide insight into the physiology and metabolism driving the high fillet yield phenotype; this information is useful for the development of genomic markers to enhance breeding strategies toward the improvement of performance traits.

<h4>Background</h4>Huntington's disease (HD) is a fatal, autosomal dominant neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in the HTT gene. While chorea is the hallmark motor symptom, HD presents with diverse psychiatric and cognitive manifestations that usually precede motor onset.<h4>Methods</h4>A 10-question online survey was distributed to 130 neurologists and neuro-geneticists from the European Huntington's Disease Network (EHDN) to identify clinical symptoms considered pathognonomic of HD and criteria for genetic testing. Responses from 52 specialists were anonymized and analysed using Microsoft Excel and SPSS 26.<h4>Results</h4>Respondents, averaging 18.4 years of experience, universally identified chorea as indicative of HD, alongside cognitive slowing, irritability, and gait abnormalities. Symptoms like neuropathy, limb weakness, and tremor were deemed inconsistent with HD. Notably, 19% of experts reported that ancillary symptoms would not deter them from recommending testing if a primary HD symptom was present. Without a family history, only chorea with or without additional symptoms was deemed sufficient for testing.<h4>Discussion</h4>The findings highlight the complexity of diagnosing HD, the importance of considering subtle psychiatric and cognitive symptoms, and the need for comprehensive patient counselling. Advances in genetic testing and therapeutic trials targeting the molecular root of HD offer hope for curative treatments.<h4>Conclusion</h4>This study underscores the growing recognition of HD's pleiotropy, the ethical considerations in testing, and the importance of clinical vigilance as patients may often first present in a non-neurological setting.

The DEK chromatin remodeling protein has oncogenic functions in breast cancers, but its functional role in normal mammary gland epithelium has remained unexplored. We developed two novel genetically engineered mouse models to study the role of Dek in normal mammary gland biology in vivo. Mammary gland-specific Dek transgenic mice developed hyperplasia and had a transcriptional profile that revealed increased expression of cell cycle, mammary stem/progenitor, and lactation-associated genes. Conversely, Dek knockout mice exhibited mammary gland functional defects resulting in dramatically reduced pup survival. Analysis of previously published scRNA-sequencing of mouse mammary glands revealed that <i>Dek</i> is most highly expressed in mammary stem cells and alveolar progenitor cells, supporting the observed phenotypes. Mechanistically, we discovered that Dek is a modifier of Ezh2 methyltransferase activity, up-regulating the levels of histone H3K27me3 to control gene transcription. Combined, this is the first report to show that Dek promotes proliferation of mammary epithelial cells via transcriptional deregulation of cell cycle genes, potentially via epigenetic mechanisms, in vivo.

Amyotrophic lateral sclerosis (ALS) involves motor neuron death due to mislocalized TDP-43. Pathologic TDP-43 associates with stress granules (SGs), and lowering the SG-associated protein ataxin-2 (ATXN2) using Atxn2-targeting antisense oligonucleotides prolongs survival in TAR4/4 sporadic ALS mice but failed in clinical trials likely due to poor target engagement. Here we show that an AAV with potent motor neuron transduction delivering Atxn2-targeting miRNAs reduces Atxn2 throughout the central nervous system at doses 40x lower than published work. In TAR4/4 mice, miAtxn2 increased survival (50%) and strength, and reduced motor neuron death, inflammation, and phosphorylated TDP-43. TAR4/4 transcriptomic dysregulation recapitulated ALS gene signatures that were rescued by miAtxn2, identifying potential therapeutic mechanisms and biomarkers. In slow progressing hemizygous mice, miAtxn2 slowed disease progression, and in ALS patient-derived lower motor neurons, our AAV vector transduced >95% of cells and potently reduced ATXN2 at MOI 4 logs lower than previously reported. These data support AAV-RNAi targeting ATXN2 as a translatable therapy for sporadic ALS.

Cellular senescence in stem cells compromises regenerative capacity, promotes chronic inflammation, and is implicated in aging. Hematopoietic stem and progenitor cells (HSPCs) are responsible for producing mature blood cells, however, how cellular senescence influences their function is largely unknown. Here, we show that JMJD3, a histone demethylase, activates cellular senescence by upregulating p16^Ink4a in competition with Polycomb group proteins, and reprograms HSPC integrity to overcome hematopoietic defects induced by replicative and oncogenic stresses. Jmjd3 deficiency does not alter global H3K27me3 levels, indicating that JMJD3 epigenetically regulates specific and limited JMJD3 targets under stress. JMJD3 deficiency also impairs stem cell potential, proper cell cycle regulation, and WNT pathway activation in HSPCs under stress. These impaired phenotypes are rescued through exogenous and retroviral introduction of p16^Ink4a. This JMJD3-p16^INK4a axis in hematopoiesis is age-dependent and is distinct from cellular senescence. Treatment with a selective JMJD3 inhibitor attenuates leukemic potential during cellular senescence. Taken together, these results demonstrate that JMJD3-p16^INK4a mediates cellular senescence and plays critical roles in the functional integrity of HSPCs under stress.

<h4>Purpose</h4>The study investigates the results of kidney stone cultures and its correlation with preoperative urine tests, stone composition and postoperative systemic inflammatory response syndrome (SIRS) in patients undergoing percutaneous nephrolithotomy (PCNL).<h4>Methods</h4>Data from 338 PCNL procedures performed between January 2018 and September 2023 at two centers in the Netherlands were included for analysis. Preoperative urine tests, kidney stone analysis and the outcome of kidney stone cultures were evaluated in addition to general patient characteristics and surgical information. Preoperative urine tests and stone cultures were concordant if they were both negative or both positive with the same microorganism. Multivariable logistic regression evaluated factors associated with positive stone cultures and their relationship to postoperative SIRS.<h4>Results</h4>Stone cultures were positive in 43% of cases and were concordant with preoperative urine tests in 66% of cases. Stone culture results identified 33 different microorganisms, extending beyond urease-positive bacteria: most commonly found were Enterococcus faecalis, Escherichia coli, and Proteus mirabilis. Positive stone cultures were found across all types of stone composition. Positive stone cultures were significantly associated with postoperative SIRS (OR 3.12, 95% CI 1.01-9.65) and a positive preoperative urine test was not (OR 1.60, 95% CI 0.56-4.53).<h4>Conclusion</h4>Stone cultures provide additional microbiological insights to preoperative urine testing. All types of stone composition can be associated with positive stone cultures, questioning the current thinking of infection vs. non-infection stones.

The induction of tissue-specific vessels in in vitro living tissue systems remains challenging. Here, we directly differentiated human pluripotent stem cells into CD32b⁺ putative liver sinusoidal progenitors by dictating developmental pathways. By devising an inverted multilayered air-liquid interface culture, hepatic endoderm, septum mesenchyme, arterial and sinusoidal quadruple progenitors self-organize to generate and sustain hepatocyte-like cells neighboured by divergent endothelial subsets composed of CD32b^lowCD31^high, LYVE1⁺STAB1⁺CD32b^highCD31^lowTHBD^-vWF^- and LYVE1^-THBD⁺vWF⁺ cells. WNT2 mediates sinusoidal-to-hepatic intercellular crosstalk potentiating hepatocyte differentiation and branched endothelial network formation. Intravital imaging reveals the iPS-cell-derived putative liver sinusoidal endothelial progenitor develops fully perfused human vessels with functional sinusoid-like features. Organoid-derived hepatocyte- and sinusoid-derived coagulation factors enable correction of in vitro clotting time with Factor V-, VIII-, IX- and XI-deficient plasma, and rescues the severe bleeding phenotype in haemophilia A mice on transplantation. Advanced organoid vascularization technology allows for interrogating key insights governing organ-specific vessel development, paving the way for coagulation disorder therapeutics.

Maturation of human pluripotent stem (hPS) cell-derived cardiomyocytes is critical for their use as a model system. Here we mimic human heart maturation pathways in the setting of hPS cell-derived cardiac organoids (hCOs). Specifically, transient activation of 5' AMP-activated protein kinase and estrogen-related receptor enhanced cardiomyocyte maturation, inducing expression of mature sarcomeric and oxidative phosphorylation proteins, and increasing metabolic capacity. hCOs generated using the directed maturation protocol (DM-hCOs) recapitulate cardiac drug responses and, when derived from calsequestrin 2 (CASQ2) and ryanodine receptor 2 (RYR2) mutant hPS cells exhibit a pro-arrhythmia phenotype. These DM-hCOs also comprise multiple cell types, which we characterize and benchmark to the human heart. Modeling of cardiomyopathy caused by a desmoplakin (DSP) mutation resulted in fibrosis and cardiac dysfunction and led to identifying the bromodomain and extra-terminal inhibitor INCB054329 as a drug mitigating the desmoplakin-related functional defect. These findings establish DM-hCOs as a versatile platform for applications in cardiac biology, disease and drug screening.

Bipolar disorder is a heritable mental illness with complex etiology. While the largest published genome-wide association study identified 64 bipolar disorder risk loci, the causal SNPs and genes within these loci remain unknown. We applied a suite of statistical and functional fine-mapping methods to these loci and prioritized 17 likely causal SNPs for bipolar disorder. We mapped these SNPs to genes and investigated their likely functional consequences by integrating variant annotations, brain cell-type epigenomic annotations, brain quantitative trait loci and results from rare variant exome sequencing in bipolar disorder. Convergent lines of evidence supported the roles of genes involved in neurotransmission and neurodevelopment, including SCN2A, TRANK1, DCLK3, INSYN2B, SYNE1, THSD7A, CACNA1B, TUBBP5, FKBP2, RASGRP1, FURIN, FES, MED24 and THRA among others in bipolar disorder. These represent promising candidates for functional experiments to understand biological mechanisms and therapeutic potential. Additionally, we demonstrated that fine-mapping effect sizes can improve performance of bipolar disorder polygenic risk scores across diverse populations and present a high-throughput fine-mapping pipeline.

Fibrosis is a major complication of Crohn's disease (CD) marked by excess deposition of extracellular matrix, leading to stricturing and functional impairment. As mechanistic characterization and therapeutic options are lacking, we paired single-cell and spatial transcriptomics in 61 samples from 21 patients with CD and 10 patients without inflammatory bowel disease (IBD). Intestinal strictures were characterized by increased immune cells, including IgG⁺ plasma cells, CCR7-hi CD4⁺ T cells and inflammatory fibroblasts. Spatial transcriptomics showed that key subsets colocalize within diseased tissues and identified additional populations such as interstitial cells of Cajal and enteric neurons. Furthermore, we mapped gene expression onto intestinal biogeography, finding that known genetic risk loci are enriched within discrete spatial modules, defined by the presence of inflammatory fibroblasts and lymphoid follicles. Altogether, our datasets chart the key transcriptomic and cellular networks in stricturing CD and highlight the spatial organization of multicellular genetic risk factors.

<h4>Background</h4>Elderly patients have an impaired functional state and multiple comorbidities, resulting in poor postoperative rehabilitation ability and high rates of disability and mortality. However, little evidence exists on mortality predictors for geriatric hip fractures within the context of the multidisciplinary team co-management model. This study aimed to investigate the incidence and explore preoperative indicators of 1-year mortality following hip fractures in the elderly under this model.<h4>Methods</h4>A total of 439 elderly patients (130 men and 309 women) surgically treated for hip fractures under the multidisciplinary team co-management model between January 2018 and June 2021were included. Data regarding demographics, health state-related variables, injury- and admission-related variables, and preoperative laboratory test results were collected from medical records. Univariate and multivariate logistic regression analyses were used to identify preoperative indicators for 1-year mortality.<h4>Results</h4>A total of 49 patients died within 1 year of hip fracture surgery between January 2018 and June 2021, with an accumulated mortality rate of 11.16%. In univariate analysis, 14 items were found to be significant. In the multivariable logistic regression model, age >85 years, body mass index <21.0 kg/m², time from injury to admission >9.5 h, preoperative haemoglobin <117 g/L, serum albumin <33.9 g/L, lactate dehydrogenase >292 U/L, and blood urea nitrogen >8.5 mmol/L were the independent preoperative indicators for 1-year mortality after surgery in elderly patients with hip fracture under the multidisciplinary team co-management model.<h4>Conclusions</h4>This study establishes a novel set of preoperative predictors for 1-year mortality in geriatric hip fracture patients managed under an MDT model, distinct from previous investigations focusing on postoperative interventions. The identified indicators enable early risk stratification, facilitating timely preoperative optimization. These findings underscore the prognostic value of integrating clinical and biochemical markers before surgery, warranting validation in multicenter prospective studies. Further prospective studies should be conducted to elucidate these associations and assess the effectiveness of targeted measures.

<h4>Background and objectives</h4>Tandem repeats (TRs) are DNA regions of tandemly repeated nucleotide motifs. Their pathogenic expansions cause various, mainly neurologic, diseases.<h4>Methods</h4>We analyzed 65 TR loci using ExpansionHunter in individuals who underwent short-read whole-exome sequencing (WES) or whole-genome sequencing (WGS) for the diagnosis of a rare neurologic condition.<h4>Results</h4>Of 1,106 proband samples (1,053 WES, 53 WGS), we detected 232 TR expansions in the intermediate or pathogenic range in 18.7% (207/1,106). However, 51 TR expansions were revised as false positives (FPs) and 83 as nondisease-causing. Of the 98 disease-causing TR expansions, 5 were classified as causal hemizygous or heterozygous TR expansions associated with X-linked recessive (XLR) or autosomal dominant (AD) neurologic disorders in 5 probands (0.5%). The low incidence is due to the fact that individuals with typical clinical symptoms (spinocerebellar ataxia) were tested for TR expansion by conventional laboratory methods. Only 1 proband with clinical suspicion of spinal and bulbar muscular atrophy was fully explained by TR expansion in the <i>AR</i> gene, and in 4 others, we hypothesize the possible involvement of 2 different neurologic diseases. Another 82 causal hemizygous or heterozygous TR expansions associated with XLR or AD non-neurologic diseases (secondary findings) were identified in 81 probands (7.3%), of which 70 expansions in <i>TCF4</i> were associated with Fuchs endothelial corneal dystrophy, a common eye disease in older patients. Finally, we detected 11 heterozygous TR expansions for XLR and autosomal recessive (AR) diseases in 11 probands who had no clinical symptoms of the associated TR disease.<h4>Discussion</h4>The unexpectedly high detection rate (18.7%) of TR expansions necessitates the filtration of FPs and nondisease-causing expansions, thereby underscoring the necessity of visual inspection of ExpansionHunter results. The study demonstrated that both WES and WGS diagnostics can benefit from TR expansion analysis. The secondary findings indicate that the previously published pathogenic ranges of TR expansions in <i>RUNX2</i> and <i>ZIC3</i> warrant further investigation.

<b>Background:</b> Eating disorders and cannabinoid hyperemesis syndrome are increasingly common causes of nausea, vomiting, and weight loss in adolescent females. Acute intermittent porphyria (AIP) is rare but has considerable pathophysiological overlap with these conditions and requires a high index of suspicion. <b>Purpose and Basic Procedures:</b> We present the case of a 15-year-old girl who presented with nausea, vomiting, and decreased appetite in the context of cannabis use and disordered eating. She was initially discharged from the emergency department but returned the next day experiencing seizures and altered mental status. Medical workup revealed AIP, and she responded well to the appropriate treatment. <b>Main Findings:</b> To date, no literature exists about the overlap between cannabinoid hyperemesis syndrome and AIP, although they often present with similar features. There is scant information about the interplay between AIP and disordered eating. As our case report shows, an AIP diagnosis could be delayed by misattribution of presenting symptoms to cannabis use or disordered eating. <b>Principal Conclusion:</b> AIP is a rare but highly treatable cause of nausea, vomiting, and altered mental status in adolescents. Due to its symptomatologic overlap with more common conditions like cannabinoid hyperemesis syndrome and eating disorders, it is easily missed. Thus, a high index of suspicion is required to obtain an AIP diagnosis and initiate treatment.

<h4>Background</h4>Crotonylation, an emerging epigenetic modification, has been implicated in various biological processes, including tumor progression. However, its role in glioma remains poorly understood. This study aims to investigate the prognostic and therapeutic implications of crotonylation-associated genes in glioma.<h4>Methods</h4>Crotonylation levels were assessed by IHC in glioma tissues of varying grades. Key crotonylation-associated genes were identified and analyzed across five glioma datasets. A prognostic risk score was developed using machine learning algorithms and validated in multiple cohorts. Genomic alterations, immune landscapes, and therapeutic responses were examined in relation to the risk score. Single-cell dataset GSE131928 was analyzed to explore the relationship between the risk score and immune cell infiltration. After crotonate treatment of T98G cells, ChIP-seq and qPCR were performed to investigate the effect of crotonylation on gene expression. Finally, PD-1 and GZMB expression levels were assessed in glioma tissues with varying crotonylation levels.<h4>Results</h4>Crotonylation levels were negatively correlated with glioma grade. Crotonylation-related genes stratified patients into two subtypes with distinct overall survival outcomes. High-risk patients exhibited increased somatic mutations, specific copy number variations, and an immunosuppressive tumor microenvironment. The risk score correlated positively with TIDE scores, indicating resistance to immune checkpoint blockade therapy. Single-cell analysis revealed a positive association between the risk score and TAM infiltration. Candidate therapeutic agents tailored for high- and low-risk groups were identified. ChIP-seq and qPCR demonstrated that reduced crotonylation suppressed <i>CXCL1</i> expression and promoted <i>GZMB</i> expression in the glioma microenvironment.<h4>Conclusion</h4>Crotonylation-associated genes play a pivotal role in glioma progression and prognosis. The risk score provides a robust tool for patient stratification and treatment guidance, underscoring the importance of crotonylation in glioma biology and its potential as a therapeutic target.

<h4>Background</h4>Huntington's Disease (HD) remains without disease-modifying treatments, with existing therapies primarily targeting chorea symptoms and offering limited benefits. This study aims to identify druggable genes and potential biomarkers for HD, focusing on using RNA-Seq analysis to uncover molecular targets and improve clinical trial outcomes.<h4>Methods</h4>We reanalyzed transcriptomic data from six independent studies comparing cortex samples of HD patients and healthy controls. The Propensity Score Matching (PSM) algorithm was applied to match cases and controls by age. Differential expression analysis (DEA) coupled with machine learning algorithms were coupled to identify differentially expressed genes (DEGs) and potential biomarkers in HD.<h4>Results</h4>Our analysis identified 5834 DEGs, including 394 putative druggable genes involved in processes like neuroinflammation, metal ion dysregulation, and blood-brain barrier dysfunction. These genes' expression levels correlated with CAG repeat length, disease onset, and progression. We also identified FTH1 as a promising biomarker for HD, with its expression downregulated in the prefrontal cortex and upregulated in peripheral blood in a CAG repeat-dependent manner.<h4>Conclusions</h4>This study highlights the potential of FTH1 as both a biomarker and a therapeutic target for HD. Advanced bioinformatics approaches like RNA-Seq and PSM are crucial for uncovering novel targets in HD, paving the way for better therapeutic interventions and improved clinical trial outcomes. Further validation of FTH1's role is needed to confirm its utility in HD.

Infertility is a growing global phenomenon affecting millions of individuals and is characterized by multifactorial causes, including both lifestyle and environmental factors. These include smoking, chronic exposure to environmental pollutants, stress, excessive caffeine or alcohol intake, drug use, improper eating habits and physical inactivity. The potential to modify these behaviors has gained increasing interest due to its impact on reproductive health and its role in mitigating infertility. Preconception counseling has also emerged as a fundamental strategy, providing education and risk assessments to improve pregnancy outcomes. Among lifestyle factors, nutrition, body composition and physical activity significantly influence female fertility, emphasizing the strong connection between metabolism and reproductive function. Supplementation with anti-inflammatory nutrients, such as omega-3 polyunsaturated fatty acids (<i>n</i>-3 PUFAs), a key component of the Mediterranean diet, may offer benefits for female fertility, partially through the modulation of gene expression in reproductive tissues. However, the specific mechanisms linking diet and fertility remain unclear. The primary objective of this review is to explore how the modification of selected lifestyle factors, with particular reference to dietary habits, may positively influence the female reproductive system and improve fertility and pregnancy-related outcomes.

Adenylyl cyclases (ACs) are key regulators of cyclic adenosine monophosphate (cAMP) signaling-a pathway critical for neuroregeneration, synaptic plasticity, and neuronal survival. In both the central and peripheral nervous systems, injury-induced activation of ACs promotes axonal outgrowth and functional recovery through the stimulation of protein kinase A (PKA), exchange proteins directly activated by cAMP (Epac), and cAMP-response element-binding protein (CREB). Among the various AC isoforms, calcium-sensitive AC1, AC8, and AC5, as well as bicarbonate-responsive soluble AC (sAC), have emerged as crucial mediators of neuroplasticity and axon regeneration. These isoforms coordinate diverse cellular responses-including gene transcription, cytoskeletal remodeling, and neurotransmitter release-to metabolic, synaptic, and injury-related signals. Dysregulation of AC activity has been implicated in the pathophysiology of neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis, as well as in chronic pain syndromes. Pharmacological modulation of cAMP levels through AC activation, phosphodiesterase (PDE) inhibition, or pituitary adenylyl cyclase-activating polypeptide (PACAP) receptor signaling has shown therapeutic promise in preclinical models by enhancing neurogenesis, remyelination, and synaptic repair. Conversely, targeted inhibition of specific AC isoforms, particularly AC1, has demonstrated efficacy in reducing maladaptive plasticity and neuropathic pain. This review highlights the diverse roles of ACs in neuronal function and injury response and discusses emerging strategies for their therapeutic targeting.

Neurology research largely utilizes rat brains due to their structural and functional similarities to humans, making them valuable models for studying various neurological conditions. There is growing interest in investigating diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), cognition, and other mental health-related disorders. This has created a need for a comprehensive, combined, and easy-to-follow method to isolate serum, cerebrospinal fluid (CSF), and hippocampal neurons. The hippocampus, responsible for learning and memory, is affected by various neurological and psychiatric disorders. However, obtaining samples like CSF and hippocampal neurons is challenging, especially from small animals like rats. Currently, there is no efficient method for isolating these samples altogether from a single animal, and its use in downstream applications has not been thoroughly tested. We have developed a comprehensive and streamlined method for isolating serum, CSF, and hippocampal neurons from a single animal, suitable for downstream applications such as proteomics and biomarker research. This method involves using high-speed centrifugation instruments and density gradient centrifugation, which are easy to follow. The isolated proteins were identified through mass spectrometry. Our method has been successfully tested for high-throughput applications with small sample volumes, demonstrating its clinical utility. With our simplified approach, proteins in serum, CSF, and neural cells can be studied simultaneously. The method achieves ease of use, cost-effectiveness, and reproducibility, thereby facilitating a better understanding of neurological disorders.

<h4>Background</h4>Familial adenomatous polyposis (FAP) is regarded as a precancerous stage of colon adenocarcinoma (COAD). COAD concurrent with FAP is quite rare in colorectal cancer screening. In order to create a new COAD prognostic prediction model and to shed light on the landscape of the tumor immune microenvironment in COAD, we examined differentially expressed genes (DEGs) between COAD and FAP in this study.<h4>Methods</h4>DEGs between COAD and FAP were identified using proteomic technology. Expression matrix data for COAD were obtained from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases. Using multivariate and least absolute shrinkage and selection operator (LASSO) regression analyses, a prognostic risk model for COAD concurrent with FAP (COAD-FAP) was created. In the training and validation sets, the efficacy of the risk model was confirmed, respectively. A systematic analysis was conducted on the relationships between the prognostic signature and immunological score, tumor immune cell infiltration, and immune checkpoints.<h4>Results</h4>A survival risk model was constructed using four prognostic genes: fatty acid-binding protein 4 (<i>FABP4</i>), cysteine-rich protein 2 (<i>CSRP2</i>), leucine-rich repeat containing helicase 4 (<i>LRCH4</i>), and tissue inhibitor of metalloproteinases 1 (<i>TIMP1</i>). The training and testing datasets both provided validation for this model. It was verified by Kaplan-Meier curves and time-dependent receiver operating characteristic (ROC) curve analysis that the prognosis of COAD could be reliably predicted by these four COAD-FAP risk signatures. The risk score was positively correlated with immune score and immune cell infiltration levels, according to the correlation analysis.<h4>Conclusions</h4>These findings imply that the four COAD-FAP risk signatures could be a practical tool for forecasting prognostic risk, assessing immunotherapy efficacy, and personalizing customized treatment choices for COAD patients.

<h4>Background</h4>Prostate cancer (PC) remains one of the leading causes of cancer-related mortality, necessitating further research into novel prognostic biomarkers and therapeutic targets. Long noncoding RNA second chromosome locus-associated with prostate-1 (SChLAP1) plays a crucial role in the aggressiveness of PC; however, its precise mechanism remains unclear. This study aimed to investigate the role of SChLAP1 in PC metastasis and apoptosis, with a particular focus on its interaction with miR-101.<h4>Methods</h4>The expression levels of SChLAP1 were analyzed by quantitative polymerase chain reaction (qPCR) and fluorescence in situ hybridization (FISH) in 42 clinical specimens, including 21 PC tissues and their corresponding adjacent normal tissues, as well as in PC cell lines (PC-3, DU145, and LNCaP). Functional assays were performed using lentivirus-mediated knockdown and overexpression of SChLAP1 and miR-101-5p. Cell proliferation, invasion, apoptosis, and autophagy were assessed via Cell Counting Kit-8 assays, Transwell migration assays, flow cytometry, and Western blot analysis. Bioinformatics analysis and complementary expression experiments were used to validate the interaction between SChLAP1 and miR-101-5p. An <i>in vivo</i> xenograft model was established by subcutaneously implanting PC-3 cells into nude mice to evaluate tumor growth.<h4>Results</h4>SChLAP1 was significantly upregulated in PC tissues and was correlated with higher Gleason scores (P<0.05). Knockdown of SChLAP1 suppressed PC-3 cell proliferation, invasion, and cell cycle progression while promoting apoptosis through MMP-9/Bcl-2 downregulation and caspase-3 activation. SChLAP1 functioned as a cytoplasmic competing endogenous RNA (ceRNA) by directly binding to miR-101-5p. Overexpression of miR-101-5p inhibited metastasis and induced apoptosis but had no significant effect on autophagy. <i>In vivo</i>, both SChLAP1 knockdown and miR-101-5p overexpression significantly reduced tumor volume.<h4>Conclusions</h4>SChLAP1 promotes PC progression by regulating metastasis and apoptosis via the miR-101-5p axis. The SChLAP1/miR-101-5p signaling pathway represents a novel diagnostic and therapeutic target, with potential implications for improving prognostic assessment and treatment strategies for advanced PC. Further clinical studies are warranted to evaluate its therapeutic potential in clinical settings.

Colorectal cancer (CRC) is a major global health concern, particularly in Western countries where there is high consumption of processed food. Gut microbiota, intestinal inflammation, and autophagy play pivotal roles in CRC initiation and progression. Probiotics and probiotic metabolites (particularly short-chain fatty acids) have emerged as potential preventive and adjuvant therapeutics by restoring a balanced gut microbiota, dampening inflammation, stimulating immune response, and improving barrier integrity and intestinal epithelial homeostasis by modulating autophagy. This narrative review discusses the current evidence supporting the anti-inflammatory, immunomodulatory, and pro-autophagy effects of probiotics and their metabolites and explores their potential preventive and therapeutic applications in CRC management.

<b>Background</b>: Scyllo-inositol (SCI) is a naturally occurring carbocyclic sugar implicated in many biological processes. Studies have highlighted the potential of using SCI in Alzheimer's therapy. However, in order to fully use this compound in the treatment of neurovegetative diseases, its pharmacokinetics must be thoroughly understood. <b>Objectives</b>: We undertook the task of analyzing SCI in a Wistar rat animal model. The aim of this study was to observe the changes in SCI concentration after oral administration. <b>Methods</b>: All rats received 10 mg/kg of SCI as a solution in distilled water by oral gavage. Estimated parameters were based on the serum concentration of SCI observed in six individual rats with regard to time. <b>Results</b>: The first peak concentration appeared at 30 min for SCI. Thereafter, the serum SCI concentration increased rapidly and reached its highest level after approximately 1.5 h. There was no second peak in SCI concentration. The elimination half-life was determined to be 10.07 h and the mean residence time was 14.52 h. There were no side effects of SCI supplementation noticed during the study. <b>Conclusions</b>: Although our results present an analysis of SCI immediately after oral administration up to 48 h, further studies are necessary.

The health problem of infertility has garnered increasing attention, prompting a deeper understanding of its causes. The broad-spectrum and nonselective herbicide glufosinate ammonium (GLA) is widely used in many countries. Previous studies have demonstrated the reproductive toxicity of GLA, but its potential toxic mechanisms remain unclear. Here, mice, Sertoli cells, and Leydig cells were used to create GLA preconditioning models. Results showed that GLA exposure caused morphological and functional damage of sperm. Concurrently, our study revealed that GLA, similar to Erastin, could induce ferroptosis in Sertoli and Leydig cells, as indicated by the dose-dependent increases of intracellular iron levels, lipid peroxidation, and cell death. Additionally, both the lipid ROS scavenger Fer and the iron chelator deferiprone were found to mitigate GLA-induced cell death. Intriguingly, our findings suggested that GLA-induced ferroptosis was dependent on autophagy, as the use of pharmacological inhibitors (3-methyladenine, chloroquine, and bafilomycin A1) or autophagy-related gene 5 gene knockout markedly reduced ferroptosis induced by GLA. We also demonstrated that nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy, which involves the autophagic degradation of the primary intracellular iron storage protein ferritin, is essential for GLA-induced ferroptosis by showing that NCOA4 knockdown decreased intracellular iron levels and attenuated lipid peroxidation, eventually alleviating GLA-induced cell death. Moreover, we observed that inhibition of the AMP-activated protein kinase-Unc-51-like kinase 1 (AMPK-ULK1) pathway activity by knockdown of AMPK expression markedly reduced the mitochondrial reactive oxygen species (mtROS) level and alleviated GLA-induced ferroptosis. Collectively, GLA induced excessive mtROS production through activation of the AMPK-ULK1 pathway, triggering excessive autophagy that ultimately led to ferroptosis via NCOA4-mediated ferritinophagy.

<h4>ABSTRACT</h4> Huntington’s disease (HD) is a progressive neurodegenerative disorder with no approved therapies. Two major molecular drivers—somatic expansion of inherited CAG repeats and toxic mutant HTT (mHTT) variants—lead to neuronal dysfunction. Despite multiple trials, HTT-lowering strategies have not shown meaningful clinical benefit. Using therapeutic divalent siRNAs, we assessed the long-term impact of silencing MSH3 (a key regulator of somatic expansion), HTT, or both. In Q111 HD mice (>110 CAGs), which exhibit robust expansion, mHTT inclusions, and transcriptional dysregulation by 12 months, long-term MSH3 silencing blocked expansion, reduced inclusions, and reversed gene expression changes. HTT silencing alone had limited effect, but combined MSH3/HTT targeting synergistically eliminated inclusions and restored transcriptomic profiles. Parallel treatment in wild-type mice showed no toxicity, supporting the safety of long-term intervention. These findings position somatic expansion as a promising therapeutic target and demonstrate the potential of RNAi-based co-silencing of MSH3 and HTT as a disease-modifying strategy for HD.

Huntington’s disease (HD) is a monogenic autosomal dominant neurodegenerative disorder caused by a CAG repeat expansion in the first exon of the HTT gene, yielding a gain-of-toxic-function mutant Huntingtin protein mHTT. CRISPR/Cas9 is a potentially powerful therapeutic tool for treating HD by eliminating mutant HTT (m HTT ) gene. We developed a specific SaCas9 guide RNA to target human m HTT , and a self-inactivating gene editing system that abolishes SaCas9 after a short transient expression for high gene editing efficiency and maximal safety to prevent off-target effects. Both conventional and the new self-inactivating gene editing systems achieved successful elimination of m HTT gene, 60-90% mHTT protein and 90% of mHTT aggregation in BAC226Q HD mouse brains, which resulted in significant long-term rescue of neural pathology, motor deficits, weight loss and shortened lifespan. These beneficial effects were observed when gene editing was applied before, at and well after the on-set of pathological and behavioral abnormalities. These proof-of-concept data demonstrate that gene editing can be a highly effective therapeutic approach for HD and other inherited neurodegenerative diseases. <h4>One Sentence Summary</h4> Self-inactivating CRISPR for mutant huntingtin in HD mice achieved long-term rescue of neural pathology, motor deficits, weight loss and survival.

Most genetic variants associated with complex diseases lie in non-coding regions, complicating efforts to identify effector genes and relevant cell types. Here, we map cis-eQTLs across 2.2 million single cells from blood and intestinal biopsies of 421 individuals, including 125 with inflammatory bowel disease (IBD). Cell-type-level eQTLs were more distal to transcription start sites, enriched in enhancers, less likely to regulate the nearest gene, and over two-fold more likely to colocalise with IBD GWAS loci than eQTLs detected at tissue-level resolution. We nominate effector genes at over half of known IBD loci, including MAML2 , PSEN2, and ZMIZ1 in myeloid cells, implicating reduced Notch signalling in intestinal immune dysfunction. We also identify Wnt regulated genes, including MYC , in epithelial stem and progenitor cells, suggesting that impaired renewal contributes to barrier breakdown. Our results provide a mechanistic map linking genetic risk to specific genes and cell types in IBD, and a framework for effector gene discovery in complex disease.

<title>Abstract</title> <p><bold>Background</bold> Several studies have demonstrated that impaired metabolism of branched chain amino acids (BCAAs) is related to cancer progression. However, the specific mechanisms underlying BCAA metabolism in head and neck squamous cell carcinoma (HNSCC) remain to be explored. The aim of this study was to identify prognostic genes associated with BCAA metabolism in HNSCC and to elucidate their functional mechanisms. <bold>Methods</bold> The HNSCC related datasets (TCGA-HNSCC, GSE65858 and GSE140042) were enrolled in this study. Candidate genes were acquired by overlapping differentially expressed genes form differential expression analysis and key module genes connected with BCAA-metabolism related genes (BCAA-MRGs) scores from weighted gene co-expression network analysis. Subsequently, prognostic genes were obtained to construct the risk model through univariate Cox regression analysis, proportional hazards hypothesis test, and least absolute shrinkage and selection operator regression analysis selected in sequence. Afterwards, independent prognostic analysis, enrichment analysis and immune microenvironment analysis were performed. Furthermore, the expression changes of prognostic genes at the cellular level were assessed through single-cell RNA sequencing (scRNA-seq) data analysis and pseudo-time analysis. Additionally, RT-qPCR was used to confirm the expression levels of prognostic genes in HNSCC tissues. <bold>Results</bold> SMS, PRDX6, GSTO1, and ADA were determined as prognostic genes to create the risk model. The HNSCC samples were divided into high-risk group (HRG) and low-risk group (LRG), with LRG demonstrating significantly higher survival rates compared to the HRG. Furthermore, the nomogram model constructed using risk score and age had an excellent predictive ability for HNSCC patients. Enrichment analysis revealed that ‘pentose phosphate pathway’ and ‘fructose and mannose metabolism’ were significantly associated with HNSCC progression. At the same time, we also found that the level of infiltration of 20 immune cells (plasmacytoid dendritic cells, mast cells, and T follicle helper cells) and the expression of 10 immune checkpoints (CD276, CD27, and CD40) differed between the HRG and the LRG. Additionally, epithelial cells were selected as key cells due to higher expression of prognostic genes. Importantly, the trend of prognostic gene expression varied with different stages of cell differentiation. Through RT-qPCR experiment, SMS, GSTO1, and ADA all expressed highly in the tumor group, but PRDX6 had not remarkably difference between tumor and normal groups. <bold>Conclusion</bold> In summary, we pinpointed four genes-SMS, PRDX6, GSTO1, and ADA-linked to the prognosis of HNSCC within the context of BCAA metabolism. Subsequently, we developed a risk model. This model offers a novel reference for prognostic assessment and treatment strategies tailored to HNSCC patients.</p>

<h4>Background</h4>Persistent elevation of aspartate aminotransferase (AST) is commonly indicative of liver injury or disease, but isolated AST elevation without concurrent alanine aminotransferase (ALT) increase is rare and difficult to diagnose. While AST is non-specific and found in various tissues, its isolated elevation is due to less common conditions, such as macro-AST, where AST binds with immunoglobulins creating a high-molecular-weight complex that affects serum activity.<h4>Case description</h4>A 68-year-old female with a history of high-grade serous ovarian cancer (HGSOC) who had persistent isolated AST elevation for several years. Evaluations including physical exams, imaging, and routine liver function tests showed no evidence of hepatic or muscular disease. The polyethylene glycol (PEG) precipitation significantly reduced serum AST activity, confirming the presence of the macro-enzyme form of AST (macro-AST).<h4>Conclusion</h4>This case highlights the rare and novel occurrence of macro-AST in a patient with ovarian cancer. It emphasizes the importance of considering macro-AST in the differential diagnosis of isolated AST elevation, particularly in patients without clear evidence of liver or muscular disease. Recognizing this benign condition can prevent unnecessary diagnostic procedures and anxiety.

The phenotype of occult hepatitis C virus (HCV) infection (OCI) in anti-HCV treatment-naive patients remains elusive. A prospective 3-year cohort study of 218 anti-HCV treatment-naive patients was conducted. OCI was defined as positive HCV RNA in peripheral blood mononuclear cells (PBMCs) of patients with negative serum HCV RNA. Among the 128 HCV antibody (Ab)-positive patients, 15 (11.7%) had OCI; higher HCV Ab titers were noted in patients with OCI than in those without OCI (84.25 ± 64.76 vs 46.60 ± 46.07 ng/mL, <i>P</i> = 0.006), and the HCV Ab titer was a marker of OCI (cutoff: >53.2 ng/mL). The 3-year cumulative incidence (CI) of cardiovascular events was greater in patients with OCI than in those without OCI (84% vs 46.4%, <i>P</i> = 0.043). Among the 90 HCV Ab-negative patients, 5 (5.6%) had OCI; the alanine aminotransferase (ALT) levels were greater in patients with OCI than in those without OCI (114.4 ± 71.6 vs 67.2 ± 87.4 U/L, <i>P</i> = 0.035). A greater CI of autoimmune disease was noted in patients with OCI than in those without OCI (20% vs 1.2%, <i>P</i> = 0.006). Among anti-HCV treatment-naive patients, the prevalence rates of OCI were 11.7% in HCV Ab-positive patients and 5.6% in HCV Ab-negative patients. High HCV Ab titers in HCV Ab-positive patients require caution regarding OCI and cardiovascular events, and cryptogenic hepatitis warrants suspicion of OCI and autoimmune diseases.IMPORTANCEAmong anti-HCV treatment-naive patients, the prevalence rates of OCI were 11.7% in HCV Ab-positive patients and 5.6% in HCV Ab-negative patients. High HCV Ab titers (cutoff: >53.2 ng/mL) in HCV Ab-positive patients require caution regarding OCI and cardiovascular events, and cryptogenic hepatitis warrants suspicion of OCI and autoimmune diseases.

BK polyomavirus (BKPyV) is a non-enveloped, double-stranded, circular DNA virus that is a member of the <i>Polyomaviridae</i> family. The BKPyV genome is divided into three regions, including the non-coding control region (NCCR), the early region, and the late region. BKPyV has one of the highest mutation rates among DNA viruses, and four genotypes have been identified based on amino acid variation within the VP1 region. Mutations within the NCCR have been noted, and this region exhibits hypervariability. Here, we show that many of the point mutations observed within the NCCR are genotype-associated, termed genotype-associated polymorphisms (GAPs). These GAPs correlate with regions of hypervariability, are inherent to their genotype, and can be used to genotype clinical strains, separate from other genomic regions. We also show that these GAPs fall within predicted transcription factor binding sites and therefore provide targets for further functional studies.IMPORTANCEBK Polyomavirus (BKPyV) is the cause of hemorrhagic cystitis in hematopoietic cell transplant recipients and BKPyV-associated nephropathy in renal transplant recipients and thus is an important determinant of transplant outcome. The viral mechanisms leading to disease manifestation remain to be thoroughly explored, but viral genetic variation has emerged as an area of interest. Understanding genomic diversity between and within BKPyV genotypes can provide sites of interest that may ultimately improve screening strategies and provide insights into the viral factors that contribute to disease.

Ultra-high-resolution Fourier transform ion cyclotron resonance mass spectrometry (UHR FT-ICR MS) for top-down proteomics has shown the potential to resolve proteoforms and splice variants, particularly those with post-translational modifications. Here, we integrate trapped ion mobility and mass selection in tandem with ultraviolet photodissociation (UVPD) followed by FT-ICR MS measurements. The proposed method using mobility/mass-selected UVPD before FT-ICR MS allows for high protein sequence coverage and PTM localization with high mass accuracy (<1 ppm) and a better duty cycle (2×). When applied to the analysis of a bovine histone mixture, characteristic UVPD <i>a</i>/<i>b</i>/<i>c</i>/<i>x</i>/<i>y</i>/<i>z</i> ions led to the annotation of 51 proteoforms from H2B, H2A, and H4 core histones with high sequence coverages (up to 77%). Histone variants and PTM combinations, including acetylation, mono-, di-, and trimethylation, and phosphorylation, identified at the top-down level were confirmed using bottom-up analysis. This work provides the foundation for effective mobility and mass preselection of precursor ions and better annotation and spectral decongestion of UVPD fragments from protein mixtures, with general applicability for top-down proteoform analysis with minimal sample preparation.

Neurodegenerative conditions, including Alzheimer's disease, Parkinson's disease, and Huntington's disease, result in a substantial health problem for the elderly, marked by ongoing neuronal degeneration and a deterioration in mental faculties. These disorders are frequently linked to oxidative stress, problems with mitochondria, and persistent inflammation in the brain, which worsen neuronal damage. The neurohormone melatonin, primarily secreted by the pineal gland, has gained recognition as a promising therapeutic agent due to its antioxidant, anti-inflammatory, and neuroprotective effects. Melatonin's functions extend beyond its regulation of circadian rhythms, as research has demonstrated its ability to remove free radicals, improve mitochondrial performance, and adjust immune system responses, ultimately reducing the progression of neurodegenerative disease. Research findings from preclinical and clinical trials imply that taking melatonin supplements could lead to improved cognitive abilities, slower disease progression, and an overall better quality of life for elderly individuals suffering from neurodegenerative conditions. The mechanisms through which melatonin acts, the best dosage, and its long-term effectiveness are still being researched. This review underscores the potential benefits of melatonin as a supplementary treatment for neurodegenerative disorders in older adults, stressing the necessity for additional studies to confirm its efficacy and standardize its use in treatment plans.

Prolonged sleep deprivation (Pr-SD) causes death in many species. While various mechanisms related to sleep regulation or this fatal consequence of sleep loss have been identified, the core molecular basis linking Pr-SD-induced lethality and sleep homeostasis remains unknown in mammals. A critical "point of no return (PONE)" status in Pr-SD subjects is highlighted in classic research, and characterizing PONE status could help uncover this mystery. Using a Pr-SD model and a reliable PONE status prediction method, we show that mice in PONE exhibit an inability to enter natural sleep, and significant disruptions in brain phosphoproteome, independent of deprivation time but closely linked to PONE status. Brain kinase or phosphatase dysfunction influences PONE status development and leads to corresponding sleep aberration concurrently. Daily 80-min recovery sleep significantly delays PONE onset and restores brain phosphoproteome. The harmful effects of excessive kinase activity on PONE development can be eliminated by combining recovery sleep and compensatory phosphatase expression. We conclude that sleep is crucial for maintaining brain phosphoproteome homeostasis, whose disruption may impact both Pr-SD-induced lethality and sleep regulation.

<h4>Background</h4>Lung cancer, one of the most prevalent and deadly malignant tumors, is increasingly common globally in terms of both morbidity and mortality rates. Unlike pyroptosis and necroptosis, cuproptosis is a non-apoptotic programmed cell death process. Studies have demonstrated that cuproptosis is involved in the incidence, development, and metastasis of lung cancer. However, the mechanisms underlying cuproptosis in lung cancer remain to be fully elucidated.<h4>Purpose</h4>In this work, we primarily examine the most recent research on copper homeostasis, cuproptosis mechanisms, and the connection between lung cancer and cuproptosis.<h4>Methods</h4>A comprehensive literature search was conducted using the Web of Science, PubMed, and Google Scholar databases. The search strategy employed a combination of the keywords "cuproptosis" and "lung cancer." The results encompassed publications from January 2020 to May 2025. This study aimed to gather research focusing on the relationship between lung cancer and cuproptosis, as well as the anticancer effects of drugs that induce cuproptosis.<h4>Results</h4>Our research indicates that lung cancer patients experience disruptions in copper ion homeostasis, as well as inhibition of cuproptosis, which points to a potentially effective therapeutic avenue through the induction of cuproptosis in lung cancer cells. This paper provides an overview of the mechanisms that highlight proteins associated with copper homeostasis and genes that play a role in cuproptosis in the lung cancer context. The modulation of proteins such as CTR1, ATOX1, FDX1, CCS, COX17, ATP7A, and ATP7B, in addition to LIPT1 and various long non-coding RNAs (AL691432.2, AC093010.2, AC107464.3, COLCA1, AC026471.3, LINC01833, and ITGB1-DT), along with genes such as DβH, LOXL2, SOD1, UBE2D1, and UBE2D3, can affect these mechanisms. These elements are vital for evaluating clinical interventions for lung cancer patients, predicting risks, and recognizing drug resistance. Moreover, we identified that pharmaceuticals facilitating copper ion transport (elesclomol and disulfiram), copper-chelating agents (DPEN, ATTM, and TETA), and nanoparticles engineered to transport copper are capable of effectively adjusting copper ion concentrations in lung cancer cells, subsequently inducing cuproptosis as a prospective treatment strategy for lung cancer. Furthermore, our findings suggest that lung cancer cells might counteract the harmful impacts of copper buildup by modulating proteins linked to copper metabolism, bolstering the antioxidant defense mechanisms, altering metabolic pathways, and triggering cellular stress responses and repair mechanisms.<h4>Conclusion</h4>Cuproptosis represents a novel mechanism dependent on copper that has important implications for the treatment of lung cancer. This process primarily involves the buildup of copper ions, which leads to a disruption in protein homeostasis, ultimately resulting in cell death. Additionally, the abnormal expression of crucial regulatory genes, such as FDX1 and LIPT1, along with transport proteins like CTR1 and ATP7A/B, is closely linked to the advancement of lung cancer. At present, drugs that act as carriers for copper ions (such as elesclomol and disulfiram), metal-organic frameworks based on copper, and copper chelators (including D-penicillamine and ammonium tetrathiomolybdate) have demonstrated promise in eliciting copper-mediated cell death in lung cancer cells. These discoveries suggest new potential targets and strategies for treating lung cancer, which could enhance the prognosis for patients diagnosed with the disease.

BACKGROUND: Hemoglobin metabolism disorder can result in systemic iron overload, leading to pigmentation in multiple organs. Although these disorders are often of genetic origin, the specific genes and mechanisms remain incompletely understood. Lanping black bone sheep (LP–BBS), a unique population from the high altitudes along the Hengduan Mountains in Yunnan province, exhibits hyperpigmentation in multiple tissues. Investigating the genetic and environmental factors underlying this phenotype provides a natural model to better understand hemoglobin metabolism disorder. RESULTS: LP-BBS were found to exhibit increased red blood cell counts, elevated hemoglobin levels, and systemic iron overload, evidenced by hyperpigmentation in various tissues. Histological and molecular analyses revealed that hyperpigmentation is driven by ferriheme overload, an inheritable quantitative trait influenced by both genetic variation and environmental factors. Genome-wide association studies identified FRRS1L as a candidate gene, with significant mutations in its 3′-untranslated region (3′-UTR) reducing FRRS1L expression. Functional assays demonstrated that insufficient FRRS1L expression promotes ferriheme accumulation in reticuloendothelial cells and macrophages, as confirmed in vitro using FRRS1L knockdown models. Ferriheme overload was associated with oxidative stress and systemic inflammation, causing pathological damage to critical organs such as the kidney, liver, and uterus. CONCLUSION: This study identifies FRRS1L as a key contributor to ferriheme overload through aberrant hemoglobin metabolism in LP–BBS. These findings offer new insights into the genetic basis and pathological mechanisms of iron overload disorders, providing a potential target for therapeutic intervention. Moreover, LP–BBS serves as a valuable natural model for studying hematogenous pigment disorders and their interplay with environmental factors.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder in which motor neurons (MNs) of the brain and spinal cord degenerate, leading to paralysis. Generating MNs from patient-specific induced pluripotent stem cells (iPSCs) may help elucidate early stages of disease. Here, we combined MNs from patients with early-onset disease with brain microvascular endothelial-like cells in a microfluidic device we termed spinal cord chips (SC-chips) and added media flow, which enhanced neuronal maturation and improved cellular health. Bulk transcriptomic and proteomic analyses of SC-chips revealed differences between control and ALS samples, including increased levels of neurofilaments. Single-nuclei RNA sequencing revealed the presence of two MN subpopulations and an ALS-specific dysregulation of glutamatergic and synaptic signaling. This ALS SC-chip model generates a diversity of mature MNs to better understand ALS pathology in a model that has an active blood-brain barrier-like system for future drug screening.

During mouse gastrulation, extraembryonic mesoderm (ExEM) contributes to the extraembryonic yolk sac (YS) and allantois, both of which are essential for successful gestation. Although the genetic networks coordinating intra-embryonic mesodermal subtype specification are well studied, ExEM diversification remains poorly understood. Here, we identify that embryoid body (EB) in vitro differentiation generates distinct lineages of mesodermal cells, matching YS and allantois development. Combining in vitro and in vivo mouse models, we discover that Eomesodermin (Eomes) controls the formation of YS-fated ExEM but is dispensable for allantois formation. Furthermore, simultaneous disruption of Eomes and T impedes the specification of any YS or allantois mesoderm, indicating compensatory roles for T during allantois formation upon Eomes depletion. Our study highlights previously unrecognized functional and mechanistic diversity in ExEM diversification and endothelial development and introduces a tractable EB model to dissect the signaling pathways and transcriptional networks driving the formation of key extraembryonic tissues.

<h4>Objectives</h4>Ruxolitinib is used to treat myelofibrosis, polycythemia vera, and steroid-refractory graft-versus-host disease following allogeneic stem cell transplantation. This study aimed to determine the association between ruxolitinib and adverse events by evaluating case reports published between January 2014 and March 2024 in the Japanese Adverse Drug Event Report database.<h4>Methods</h4>The signals for the ruxolitinib-adverse event association were identified using propensity score-adjusted reporting odds ratio analysis. Data obtained from the drug-gene interaction, drug signature, search tool for chemical interactions, and interaction reference index databases were used to construct a drug-gene interaction network. Functional and pathway enrichment analyses were performed using the Disease Ontology Semantic and Enrichment and ReactomePA R packages.<h4>Results</h4>The propensity score-adjusted reporting odds ratio for ruxolitinib-associated adverse events was as follows: anemia, 18.49 (95% confidence interval (CI): 16.15-21.16); myelosuppression, 4.70 (95% CI: 3.54-6.24); pancytopenia, 1.97 (95% CI: 1.23-3.16); cardiac failure, 2.29 (95% CI: 1.60-3.28); hepatic function abnormal, 1.60 (95% CI: 1.15-2.23); herpes zoster, 6.40 (95% CI: 4.35-9.41); pneumonia, 2.96 (95% CI: 2.35-3.73); renal impairment, 1.34 (95% CI: 0.94-1.90); sepsis, 5.14 (95% CI: 3.75-7.05); interstitial lung disease, 0.33 (95% CI: 0.21-0.52); deep vein thrombosis, 0.32 (95% CI: 0.07-1.44); hemorrhage, 1.99 (95% CI: 1.05-3.75). We also assessed 3015 human genes that directly or indirectly interact with ruxolitinib. The molecular complex detection plug-in of Cytoscape was used to detect 24 clusters. Several genes were enriched in the biological processes of "anemia" and "bacterial infections," identified as significant ruxolitinib-related disease terms.<h4>Conclusions</h4>This retrospective analysis using the Japanese Adverse Drug Event Report database indicated potential associations between ruxolitinib and adverse events, including anemia and bacterial infections. Future research should explore the underlying pharmacological mechanisms using functional enrichment analysis of ruxolitinib-associated genes related to blood toxicity and bacterial infections.

Fibrillogenesis of plasma fibronectin (FN) can be triggered by shear stress, even in the absence of cells. While high shear stress is known to promote FN fibril formation, recent evidence suggests that low shear conditions also contribute to FN assembly. It is likely that distinct molecular mechanisms govern fibril formation under different flow regimes. In this study, we investigated FN fibril formation under high shear (2000-5000 s^-1) and low shear (50 s^-1) conditions which correspond to 7-17.5 Pa and 0.175 Pa, respectively. Morphological analysis showed that high shear rapidly induced thick, interconnected fibrils, whereas low shear led to the gradual formation of thin, sparse fibrils. Molecular dynamic simulations indicated that shear stress alone did not unfold FN in suspension; however, surface-adsorbed FN enabled shear-driven interaction with soluble FN. Under low shear, fibrillogenesis proceeded via slow FN accumulation and weak intermolecular binding, while high shear promoted strong domain-domain interactions. Molecular docking identified top-ranked trans-binding interfaces between FNI1-5 and FNIII1-3 regions, stabilized by salt bridges (e.g., Glu92-Arg694) and hydrogen bonds involving residues such as Lys149-Gly713 and Tyr265-Ser864. These interdomain interactions provide a structural basis for shear-induced fibril assembly.

Body weight (BW) is a crucial indicator of animal growth and development, significantly influencing animal husbandry practices. Previous research has identified several genes associated with BW in certain yak breeds. However, the genetic basis of BW in Gannan yaks has not been reported. In this study, 309 yaks from six breeds across five provinces in China were sampled. This collection included 247 54-month-old female Gannan yaks, along with 20 Xizang yaks, 15 Muli yaks, 10 Pamir yaks, 9 Bazhou yaks, and 8 Zhongdian yaks. Body weight measurements were recorded for the Gannan yaks. Initial analyses of runs of homozygosity (ROH), nucleotide diversity, and linkage disequilibrium (LD) decay among the six yak breeds revealed that Gannan yaks exhibited the lowest ROH, the highest nucleotide diversity, and the fastest LD decay, indicating rich genetic diversity. Subsequently, a genome-wide association study (GWAS) identified 19 BW-related genes in the Gannan yaks, with <i>PMAIP1</i>, <i>GABBR1</i>, <i>LRPPRC</i>, and <i>PPP1R11</i> identified as key genes. Genome-wide scanning of Group 1 and Group 2 (detailed in section 2.4) identified 90 genes, and Gene Ontology (GO) analysis highlighted <i>FGF2</i>, <i>SHH</i>, and <i>WNT11</i> as significantly associated with growth, development, and metabolism. Three overlapping genes were identified between GWAS and genome-wide scans. Further analyses, including nucleotide diversity, LD analysis of significant GWAS sites, allele frequency analysis, and SNP association studies, suggested that <i>DRC1</i> and <i>SELENOI</i> are novel candidate genes for BW in Gannan yaks. These findings provide a molecular foundation for the genetic improvement of Gannan yaks.

<h4>Introduction</h4>In mammals, the ubiquitin-proteasomal pathway plays a key role in the host antiviral response by targeting viral genes for degradation. Here, E1-activating enzymes, E2-conjugating enzymes, and E3 ligases attach the ubiquitin chain to molecules to be functionally modified or destined for degradation. One specialized version of this pathway is performed by ISG15, a ubiquitin-like protein modifier that works through a process called ISGylation. In mammals, ISGylation involves specialized E1-E3 molecules and specialized mammalian ISG15 "deubiquitinases" also exist. Targeting host and viral proteins, ISG15 can inhibit the release of viral particles or hinder viral replication, thereby exerting strong antiviral effects. In Atlantic salmon, endothelial cells from the heart is a main target for infectious salmon anemia virus (ISAV).<h4>Results</h4>Here, we established a new cell line from Atlantic salmon heart tissue denoted ASH2-2, which has endothelial-like characteristics and is permissive for infection with ISAV and other salmonid viruses. We used this cell line as a model to compare the effect of recombinant interferon gamma (rIFNg) and ISAV on genes potentially involved in the ubiquitin-proteasome pathway. ASH2-2 cells have a response profile matching endothelial cells and respond quickly to ISAV infection with upregulation of viral sensors such as DHX58, MDA5, and MX transcripts. Two <i>ISG15</i> genes are strongly upregulated 48 h post-infection (p.i.) while other ubiquitin genes were unaffected. Related to the mammalian E3 ligases known to be ISGylated, phylogenetic analysis identified two additional teleost-specific HERC8 and HERC9 clusters in addition to the clade previously defined as HERC7. Duplicate genes for Atlantic salmon <i>HERC7</i> and <i>HERC9</i> are both more upregulated by virus and rIFNg at 24 h p.i. as opposed to <i>HERC8</i> genes. Early regulation of the ISGylation process is also indicated by a strong upregulation of one <i>USP18</i> duplicate already at 4 h p.i.<h4>Conclusion</h4>In conclusion, we expand the list of teleost genes potentially involved in the ubiquitin-proteasome pathway in cells from a main target organ. Our results highlight the need for functional studies to clarify the roles of these candidates.

<h4>Objective</h4>To explore the chain mediating role of environmental adaptation and coping styles between life stressors and depression among university students.<h4>Methods</h4>Twelve thousand, one hundred ninety-eight college students were investigated with Student Life Stress Inventory (SLSI), Depressive Episode Scale (DES), Environmental Adaptation Scale (EAS), and Coping Style Scale (CSS). The chain intermediary model was constructed and verified with SPSS.<h4>Results</h4>Life stressors and coping styles were positive predictors of depression, while environmental adaptation was a negative predictors. The direct impact of life stressors on depression was significant. Environmental adaptation and coping styles exhibited both independent mediating effects and a chain mediating effect.<h4>Conclusion</h4>Life stressors can directly lead to depression, and indirectly exacerbate depression through environmental adaptation and coping styles. Therefore, reducing burden, enhancing environmental adaptability and learning appropriate coping strategies can effectively prevent depression.

<h4>Background</h4>Ulcerative colitis (UC) has a complex etiology, and whether there are sex-related differences in its molecular mechanisms remains unclear. This study employed multi-omics analysis to explore sex-based differences in UC, aiming to provide support for personalized treatment.<h4>Methods</h4>The GSE36807 and GSE206171 datasets from the GEO database were grouped by sex. Data were preprocessed using the R software, and DEGs identified using the limma package and key modules of WGCNA. LASSO regression was conducted to screen hub genes, ROC curves were used to evaluate diagnostic value, CIBERSORT was used to analyze immune cell proportions, and Spearman's correlation was performed to explore associations. The single-cell dataset GSE214695 was processed using Seurat to analyze immune cell proportion differences. Histological, immunohistochemical, and metabolomic analyses were performed on the colon tissues of DSS-induced colitis model mice.<h4>Results</h4>Thirty-seven DEGs and 47 co-expression modules were identified. LASSO regression highlighted RPS4Y1 as the core gene, which was significantly upregulated in males. Females showed higher proportions of resting NK cells and M0 macrophages but a lower number of eosinophils. RPS4Y1 expression was positively correlated with resting memory CD4+ T cells and eosinophils and negatively with M0 macrophages and resting mast cells. Macrophage function exhibited sex-based disparities. Increased immune cell infiltration was observed in female colon tissues compared with that in male colon tissues. Metabolomic analysis identified 140 sex-dimorphic metabolites, with significant alterations in glutathione metabolism.<h4>Conclusion</h4>RPS4Y1 exhibits sex-specific expression in UC and plays a key role in immunomodulation. Mitochondrial energy metabolism contributes to sex-based macrophage differences, highlighting the importance of considering sex-specific mechanisms in UC diagnosis and individualized treatment.

Marine heatwaves (MHWs) have recently become more frequent, intense, and prolonged, posing significant threats to marine life and fisheries. In this study, transcriptomic analysis was employed to investigate the genes and pathways in <i>Seriola dumerili</i> that respond to MHW-induced stress at 28 °C (T28) and 32 °C (T32), using 24 °C (T24) as the control. Transcriptome sequencing revealed that 17 differentially expressed genes (DEGs) belonging to the heat shock protein (HSP) families-HSP30, HSP40, HSP70, and HSP90-were significantly upregulated under short-lasting MHW stress in the T24-4d vs. T32-4d comparison. Additionally, genes related to oxidative stress (e.g., protein disulfide isomerase family A member 6 [<i>pdia6</i>]), immune responses (e.g., interferon regulatory factor 5 [<i>irf5</i>]), and energy metabolism (e.g., hexokinase-1 [<i>hk1</i>]) were also identified. Enrichment analysis of DEGs in the T24-4d vs. T32-4d group revealed that <i>S. dumerili</i> exhibited adaptive responses to MHWs through the upregulation of HSPs and the activation of antioxidant, energy metabolism, and immune response pathways. However, in the T24-13d vs. T32-13d group, DEGs associated with these pathways were either not significantly expressed or were downregulated. These findings indicate that <i>S. dumerili</i> is unable to sustain its adaptive responses under repeated, intense MHW exposure, resulting in the disorder of its antioxidant defense system, immune suppression, and metabolic dysfunction. This study provides valuable insights into the molecular responses of <i>S. dumerili</i> to MHWs and supports the selection for thermal resistance in this species.

Global concerns about environmental pollution, poor waste management, and the rise in antimicrobial resistance due to uncontrolled antibiotic use have driven researchers to seek alternative, multifaceted solutions. Plants, animals, microorganisms, and their processing wastes serve as valuable sources of natural biopolymers and bioactive compounds. Through nanotechnology, these can be assembled into formulations with enhanced antimicrobial properties, high safety, and low toxicity. This review explores polysaccharides, including chitosan, alginate, starch, pectin, cellulose, hemicellulose, gums, carrageenan, dextran, pullulan, and hyaluronic acid, used in nanotechnology, highlighting their advantages and limitations as nanocarriers. Addressing the global urgency for alternative antimicrobials, we examined natural compounds derived from plants, microorganisms, and animals, such as phytochemicals, bacteriocins, animal antimicrobial peptides, and proteins. Focusing on their protection and retained activity, this review discusses polysaccharide-based nanoformulations with natural antimicrobials, including nanoparticles, nanoemulsions, nanocapsules, nanoplexes, and nanogels. Special emphasis is placed on strategies and formulations for the encapsulation, entrapment, and conjugation of natural compounds using polysaccharides as protective carriers and delivery systems, including a brief discussion on their future applications, prospects, and challenges in scaling up.

Duchenne muscular dystrophy (DMD) manifests as a hereditary condition that diminishes muscular strength through the progressive degeneration of structural muscle tissue, which is brought about by deficiencies in the dystrophin protein required for the integrity of muscle cells. DMD is among four different types of dystrophinopathy disorders. Current studies have established that long non-coding RNAs (lncRNAs) play a significant role in determining the trajectory and overall prognosis of chronic musculoskeletal conditions. LncRNAs are different in terms of their lengths, production mechanisms, and operational modes, but they do not produce proteins, as their primary activity is the regulation of gene expression. This research synthesizes current literature on the role of lncRNAs in the regulation of myogenesis with a specific focus on certain lncRNAs leading to DMD increments or suppressing muscle biological functions. LncRNAs modulate skeletal myogenesis gene expression, yet pathological lncRNA function is linked to various muscular diseases. Some lncRNAs directly control genes or indirectly control miRNAs with positive or negative effects on muscle cells or the development of DMD. The research findings have significantly advanced our knowledge about the regulatory function of lncRNAs on muscle growth and regeneration processes and DMD diseases.

Glioblastoma multiforme (GBM) is a highly aggressive, treatment-resistant grade IV brain tumor with poor prognosis that grows rapidly and invades surrounding tissues, complicating surgery and frequently recurring. Although the crucial role of endogenous peptides has been highlighted for several tumors, the specific peptidomic profile of GBM remains unexplored to date. This study aimed to perform a preliminary characterization of the low molecular mass proteome fraction of Cavitron Ultrasonic Surgical Aspirator (CUSA) fluid collected from different tumor zones, i.e., the core and tumor periphery of newly diagnosed (ND) and recurrent (R) GBM. The samples, pooled by tumor type and collection zone, were centrifuged through molecular cut-off filter devices to collect the non-retained fraction of the proteome <10 kDa for direct full-length LC-MS analysis. A total of 40 and 24 peptides, fragments of 32 and 18 proteins, were marked as ND and R GBM COREs, respectively, while 132 peptides, fragments of 46 precursor proteins, were identified as common and included proteins which were cancer-related or involved in GBM pathophysiology. Besides providing a preliminary overview of the unexplored peptidome of GBM, this pilot study confirms peptidomics as a promising tool to discover potential GBM biomarkers in the perspective of clinical applications increasingly oriented towards a precision medicine approach. Data are available via ProteomeXchange with the identifier PXD060807.

Zebrafish is a well-recognized model for studying human genetic disorders. Recently, we proposed the homozygous <i>cdkl5^sa21938</i> mutant zebrafish as a model of CDKL5 deficiency disorder (CDD), a developmental epileptic encephalopathy with diverse symptoms. This study aimed to explore Cdkl5-associated molecular mechanisms in zebrafish and assess their similarity to those in mammals. We conducted RNA sequencing on whole <i>cdkl5</i>^-/- zebrafish and wild-type siblings at 5 and 35 days post-fertilization (dpf) to compare their gene expression profiles. Most significant differentially expressed genes (DEGs) were related to muscle, neuronal, and visual systems which are affected in CDD. Gene Ontology analysis revealed downregulated DEGs enriched in muscle development, extracellular matrix, and actin cytoskeleton functions at both stages, while upregulated DEGs were enriched in eye development functions at 35 dpf. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed enrichment of downregulated DEGs in focal adhesion and extracellular matrix (ECM)-receptor interaction pathways at both stages. Neuronal development DEGs were mainly downregulated at both stages, while synaptic signaling DEGs were upregulated at 35 dpf. Crossing <i>cdkl5</i>^-/- mutants with the Hb9:GFP transgenic line showed fewer motor neuron cells with shorter axons compared to the wild type, which may explain the impaired motor phenotype observed in zebrafish and CDD patients. Moreover, we identified key downregulated DEGs related to cartilage development at both stages and bone development at 35 dpf, potentially explaining the skeletal defects seen in zebrafish and CDD individuals. In conclusion, Cdkl5 loss in zebrafish leads to dysregulation of genes involved in CDKL5-associated functions in mammals, providing new insights into its less studied functions and phenotypes.

Aberrant DNA methylation is a hallmark of colorectal cancer (CRC), contributing to tumor progression through the silencing of tumor suppressor genes and activation of oncogenes. Indicaxanthin (IND), a dietary betalain pigment from <i>Opuntia ficus indica</i>, has shown antiproliferative effects in CRC models, yet its epigenetic impact remains unexplored. In this study, we investigated the effects of IND on the methylome of Caco-2 cells using Reduced Representation Bisulfite Sequencing (RRBS). IND induced a global hypermethylation profile, particularly at gene promoters and CpG islands. Among the differentially methylated genes, 60% were protein-coding, and 10% encoded transcription factors, including <i>PAX5</i> and <i>TFAP4</i>, both hypermethylated at active enhancers. Functional enrichment analysis revealed pathways beyond canonical intestinal functions, suggesting altered cell identity and plasticity. Transcription factor targets (<i>SOX10</i>, <i>NFKB1</i>, <i>AHR</i>, <i>ARNT</i>) were significantly enriched among the affected genes, several of which are involved in transdifferentiation processes. Methylation changes also indicated potential reprogramming toward epithelial cell types from pulmonary or neuroectodermal origin. Moreover, IND induced selective hypomethylation of Alu elements on chromosome 21 and hypermethylation of rDNA loci, hinting at suppressed ribosomal biogenesis. Overall, these findings highlight the epigenetic remodeling potential of IND and its possible role in modulating cell fate and metabolism in CRC cells.

Hydroxyapatite (HAp) has emerged as a biomaterial of significant interest due to its intrinsic biocompatibility and structural similarity to natural bone minerals. While HAp is traditionally derived from natural sources, chemical synthesis <i>via</i> conventional methods, such as wet chemical precipitation and sol-gel processing, and newer techniques like microwave-assisted synthesis and hydrothermal methods have enabled greater control over its physicochemical properties. With the expansion of applications beyond conventional biomedical uses, recent research has concentrated on engineering nanohydroxyapatite with precisely tailored morphologies and structures. This review examines the influence of various organic modifiers on nano-HAp synthesis, highlighting how these agents modulate its crystal growth, crystallinity, surface topology, particle dimensions, and porosity. Potent chelating agents (<i>e.g.</i>, citric acid and EDTA) have been shown to yield purer, more uniform nanoparticles, whereas cationic-anionic surfactants (<i>e.g.</i>, CTAB and SDS) enhance the surface area. Modifiers such as Triton X-100, chitosan, and polyethylene glycol effectively adjust the pore size. Scientists are also investigating environmentally friendly and toxicant-free modifiers. Through summarization of insights from current literature, this review provides a comprehensive framework for selecting suitable modifiers to fabricate well-defined HAp nanomaterials for diverse applications in future studies.

Takotsubo syndrome (TS), also known as stress-induced cardiomyopathy, is classically characterized by an acute onset mimicking myocardial infarction and by distinctive transient wall motion abnormalities detectable via echocardiography, often resembling a Japanese octopus trap (the so-called "takotsubo"). The possibility that a genetic background may contribute to TS susceptibility emerged early, supported by several familial case reports. Despite a large number of investigations, no definitive genetic markers associated with TS risk have been conclusively identified. The lack of a clear Mendelian inheritance pattern suggests a multifactorial etiology and pathogenesis, likely involving complex gene-environment interactions and a polygenic background. This review analyzes the genetic variants implicated in the different functional pathways contributing to TS pathogenesis and discusses the current state of knowledge regarding its genetic underpinnings. Finally, we propose future directions for research aimed at identifying a multigene susceptibility panel that could be useful in diagnosis, prevention strategies, and the identification of novel therapeutic targets for individuals at high risk. We conclude that innovative approaches based on data-mining algorithms and nonlinear analytic methods applied to large patient datasets may be instrumental in resolving the genetic complexity of TS.

<b>Background/Objectives</b>: Wool is an important product in sheep production, but the genetic mechanisms underpinning variation in wool growth are not fully understood. Identifying the genes and genomic variants that play a role in increasing fleece weight may allow for increased selection accuracy and improved economic return to producers. <b>Methods</b>: A genome-wide association study (GWAS) was conducted to investigate genetic associations with lifetime fleece weight, average fleece weight and average post-lambing ewe weight for Rambouillet, Polypay, Suffolk and Columbia ewes (<i>N</i> = 1125). Weir-Cockerham F_ST and runs of homozygosity (ROH) analyses were conducted to improve detection of putative wool-related signatures. <b>Results</b>: Twenty-four SNPs were identified through GWAS for lifetime fleece weight, average fleece weight and average post-lambing ewe weight. Chromosomes 2 and 6 contained ROH islands in Rambouillet, and chromosomes 2, 3 and 10 contained ROH islands in Suffolk. The F_ST analysis identified 18 SNPs in proximity to 37 genes of interest. <b>Conclusions</b>: Many of the SNPs and signatures of selection reported in this study are near or within current candidate genes for wool production and wool quality, including <i>ADAR</i>, <i>KCNN3</i>, <i>NTN1</i>, <i>SETBP1</i>, <i>TP53</i> and <i>TNFSF12</i>. The significant SNPs implicated by GWAS may be used to predict ewes' potential for lifetime wool production and are suggested as candidates for further study to continue to elucidate the genetic mechanisms underlying wool production traits in United States sheep breeds.

<i>Background and Objectives</i>: Inherited thrombophilia (IT) increases the risk of adverse pregnancy outcomes, but the benefit of low-molecular-weight heparin (LMWH) prophylaxis remains debated. This study aimed evaluate the effect of LMWH by analyzing outcomes in women with IT who received LMWH versus those who did not and also compare pregnancy complication rates before and after inherited thrombophilia diagnosis. <i>Materials and Methods</i>: We conducted a retrospective case-control study including 276 pregnant women with inherited thrombophilia and prior pregnancy complications and 276 healthy pregnant controls on delivery. The main outcome was the incidence of complications: preterm rupture of membranes, oligohydramnios, fetal growth restriction, preterm delivery, stillbirth, HELLP syndrome, gestational hypertension, deep vein thrombosis, and recurrent pregnancy loss. The effect of LMWH was assessed by comparing complication rates among inherited thrombophilia patients who received therapy versus those who did not. <i>Results</i>: Women with IT were older, had higher BMI, delivered earlier, and had neonates with lower birth weight compared to controls. In current pregnancies, LMWH was associated with reduced rates of preterm delivery, fetal growth restriction, gestational hypertension, and recurrent pregnancy loss, especially in factor V Leiden carriers. LMWH had little effect on low-risk genotypes and was not independently associated with outcome reduction. <i>Conclusions</i>: LMWH prophylaxis should be reserved for high-risk women with IT. Routine use in all IT pregnancies is not justified and may cause unnecessary risks and costs. Early screening, risk stratification, and individualized care are essential to optimize outcomes.

<b>Background:</b> Ovarian cancer is a major challenge in oncology due to high mortality rates, especially in advanced stages, despite current therapeutic approaches relying on chemotherapy and surgery. The search for novel therapeutic strategies is driven by the need for more effective treatments. This study focuses on novel xanthone derivatives modified with a morpholine ring, aiming to improve anticancer efficacy. <b>Methods:</b> In silico studies were conducted using ProTox III and SwissADME databases to assess the toxicity and ADME properties of the synthesized compounds. Molecular changes in cellular stress-related genes were investigated through qPCR in two ovarian cancer cell lines (TOV-21G and SKOV-3) following treatment with the compounds. <b>Results:</b> In silico analyses predicted high gastrointestinal absorption and blood-brain barrier permeability for the derivatives. Compounds exhibited varying toxicity and metabolic profiles. qPCR revealed significant alterations in genes related to antioxidant enzymes, molecular chaperones, and xenobiotic metabolism, indicating potential mechanisms of action and cellular responses to the compounds. <b>Conclusions:</b> The study demonstrates the potential of novel xanthone derivatives as promising candidates for ovarian cancer therapy, with implications for enhancing therapeutic efficacy and addressing drug resistance. Further research is warranted to elucidate the precise mechanisms underlying the observed effects and to develop tailored treatment strategies leveraging these agents.

<h4>Background</h4> Serum IgA and IgM levels in cohorts of adults with hemochromatosis are not reported. <h4>Methods</h4> We compiled serum IgA and IgM levels at diagnosis of hemochromatosis in probands with HFE p.C282Y (rs1800562) homozygosity, investigated associations of IgA and IgM with clinical and other laboratory characteristics, and compared mean IgA and IgM of probands with combined/weighted means of published adult European cohorts not selected for hemochromatosis. <h4>Results</h4> There were 73 probands (36 men, 37 women; mean age 51 ± 13 y). Fifty probands (68.5%) had human leukocyte antigen (HLA)-A*03. Mean IgA ± standard deviation [95% confidence interval] was 2.11 ± 1.06 g/L [1.87, 2.35]. Mean IgM was 1.11 ± 0.75 g/L [0.94, 1.28]. IgM was inversely associated with age (Pearson’s r 73 = –0.2733; p = 0.019). A multiple regression on IgA revealed no significant association with other characteristics. A regression on IgM revealed one positive association (daily alcohol intake; p = 0.036) and one negative association (age; p = 0.016). Mean IgA of male and female probands and corresponding mean IgA of Europeans in two cohorts (918 men, 458 women) did not differ significantly. Mean IgM of probands was lower than the mean IgM of Europeans in four cohorts (men 1.03 ± 0.84 g/L vs. 1.35 ± 0.55 g/L (n = 1084)), respectively (p <0.001); women 1.18 ± 0.67 g/L vs. 1.57 ± 0.68 g/L (n = 622), respectively (p <0.001)). <h4>Conclusions</h4> Serum IgM levels of hemochromatosis probands with HFE p.C282Y homozygosity are positively associated with daily alcohol intake and inversely associated with age. Mean IgM levels of male and female probands are lower than those of European men and women not selected for hemochromatosis.

The global rise in dementia necessitates scalable cognitive assessments that can evolve to serve both clinical and research applications. We present the Oxford Cognitive Testing Portal (OCTAL), a remote, browser-based platform providing performance metrics for memory, attention, visuospatial and executive function domains. Four validation studies (N=1,749) confirmed cross-cultural applicability, lifespan sensitivity and clinical utility. Task performance was equivalent in English- and Chinese-speaking younger adults and mapped domain-specific ageing trajectories in mid- to late-adulthood. In a memory-clinic cohort (N=194), a 5-minute OCTAL screen distinguished patients with Alzheimer’s disease dementia from subjective cognitive decline (AUC = 0.92), matching a standard paper-based test, while a 20-minute subset surpassed this (AUC = 0.98; p = 0.04). Test-retest reliability was very good (ICC ≥ 0.79; N = 118). OCTAL enables remote assessment for large-scale research and screening, with an open, modular architecture that makes it a uniquely sustainable and evolvable tool for the research community.

No abstract available.

<h4>Introduction/aims</h4>Muscle cramps are a common symptom in amyotrophic lateral sclerosis (ALS). Ameliorating muscle cramps may improve quality of life in devastating diseases like ALS. A traditional Japanese medicine (Kampo, TJ-68) is widely prescribed in Japan for muscle cramps. However, it is not available in the USA. This study evaluated the safety, tolerability, and efficacy of TJ-68 in ALS.<h4>Methods</h4>This study was a double-blind, randomized, placebo-controlled crossover trial, consisting of four periods, conducted at three centers in the USA. Safety was evaluated using multiple measures. The primary efficacy outcome was the Visual Analog Scale for Muscle Cramps Affecting Overall Daily Activity (item #5 of the Muscle Cramp Scale (MCS)). The secondary outcomes included the remaining items of the MCS and the Clinical Global Impression of Changes (CGIC), among others. The study was planned to enroll 22 participants with ALS within 2 years.<h4>Results</h4>The enrollment was slow and was completed with 11 participants. There were no serious safety issues and TJ-68 was well tolerated. Although the primary outcome measure did not reach statistical significance (p = 0.35), several secondary measures showed significant results: MCS #1 triggering of cramps (p = 0.01), MCS #2 cramp frequency (p = 0.03), MCS Additional 1 change of motor behaviors (p = 0.02), and CGIC assessed by the evaluator (p = 0.009). Other outcome measures did not reach statistical significance.<h4>Discussion</h4>The study revealed that N-of-1 trial design can detect changes in a small sample size, and TJ-68 appeared to be safe. Larger studies are needed to confirm the efficacy of TJ-68.

Nanocomposites, are materials that incorporate nanosized particles into a matrix of standard material, have emerged as a versatile class of materials with tunable properties for a wide range of applications. Traditional fabrication approaches, including physical blending, in situ polymerization, layer-by-layer assembly, and sol-gel synthetic methods, have been widely employed to develop nanocomposites with high structural homogeneity and tailored properties. This review presents a cohesive and comprehensive overview of nanocomposite fabrication methods, spanning from conventional synthetic strategies to cutting-edge approaches such as 3D printing technologies. How 3D printing has driven innovations in nanocomposite applications, particularly in biomedicine, soft robotics, electronics, and water treatment, is explored. Additionally, key challenges in 3D-printed nanocomposite development are discussed, and emerging advancements such as 5D printing, artificial intelligence (AI)-assisted material optimization, nanoscale additive manufacturing, and closed-loop recycling systems are highlighted. By bridging traditional synthesis with cutting-edge fabrication techniques, this review aims to provide insights into the future directions of nanocomposite research and applications.

Spinal and bulbar muscular atrophy (SBMA) is a neuromuscular disease caused by an expanded CAG repeat in the androgen receptor (AR) gene. To elucidate the cell type-specific temporal gene expression in SBMA, we performed single-nucleus RNA sequencing on the spinal cords of an SBMA mouse model (AR-97Q). Among all cell types, oligodendrocytes had the highest number of differentially expressed genes before disease onset. Analysis of oligodendrocyte clusters suggested that pathways associated with cation channels and synaptic function were activated before disease onset, with increased output from oligodendrocytes to neurons in AR-97Q mice compared with wild-type mice. These changes in the early stages were abrogated at the advanced stages. An oligodendrocyte model of SBMA showed phenotypes similar to those of AR-97Q mice at early stages, such as increased transcriptional changes in synapse organization, and Ca2+ imaging of oligodendrocytes in AR-97Q mice revealed the increased Ca2+ responses. A coculture system of primary rat oligodendrocytes and neurons revealed that the mutant AR in oligodendrocytes affected the activity and synchronization of neurons. These findings suggest that dysregulated cell-to-cell communication plays a critical role in early SBMA pathology and that synaptic or ion channel-related proteins, such as contactin associated protein 2 (Cntnap2) and NALCN channel auxiliary factor 1 (Fam155a), are potential therapeutic targets for SBMA.

This study introduces a drug delivery system utilizing boronic ester-functionalized mesoporous silica nanoparticles (<b>MSNP-BA-Tf</b>) for targeted cancer therapy. By conjugating transferrin (Tf) to the MSNP surface, the system actively targets cancer cells via transferrin receptor (TfR)-mediated endocytosis. The incorporation of boronic ester linkages enables hydrogen peroxide (H₂O₂)-responsive drug release, enhancing therapeutic efficacy. In vitro experiments demonstrated that <b>MSNP-BA-Tf</b> effectively delivered doxorubicin (DOX) to cancer cells while sparing normal cells, as confirmed by fluorescence imaging and cytotoxicity assays. In vivo studies using a colon cancer xenograft model showed that <b>DOX@MSNP-BA-Tf</b> inhibited tumor growth more effectively than free DOX. These findings highlight <b>MSNP-BA-Tf</b> as a promising nanocarrier for cancer therapy, offering targeted and controlled drug delivery through active targeting and H₂O₂ responsiveness.

<h4>Background</h4>The incidence of small bowel neuroendocrine tumors (SBNETs) is steadily increasing, new therapies are urgently needed to prolong the overall survival of patients.<h4>Objective</h4>This study aimed to identify diagnostic and therapeutic candidate markers for SBNETs.<h4>Methods</h4>Expression profiles of miRNAs were collected from GSE70534, and GSE103317, that of mRNAs were collected from GSE65286. Differentially expressed genes (DEmiRs, DEmRs) were analyzed between SBNETs and controls. Enrichment and coexpression analyses were carried out for DEmRs. XGBoost algorithm was used to screening feature miRNAs. Module genes in SBNETs-related pathways were selected to construct regulated network for feature miRNAs. Drug targeting prediction and immune environment evaluation were identified.<h4>Results</h4>A total of 57 common DEmiRs with the same direction of expression were identified. Hsa - miR - 375, hsa - miR - 107, hsa - miR - 1180, hsa - miR - 330 - 3p, and hsa - miR - 328 were identified as feature miRNAs. Among the target genes of feature miRNAs, PDPK1 was the correlation between PDPK1 and the target of Lutetium-177 (177Lu)-DOTATATE was the largest, which were regulated by miR - 375. Additionally, PDPK1 showed correlations with eosinophils, cytotoxic cells, and checkpoints in SBNETs.<h4>Conclusions</h4>Five feature miRNAs may have a good diagnostic role for the SBNETs. MiR - 375 regulated PDPK1 may serve as an effective therapeutic candidate marker for SBNETs.

We identified novel Molecular subtypes according to the expression of fibrosis-related genes (FRGs) and constructed a prognostic model using different expression genes (DEGs) for patients with Hepatocellular carcinoma (HCC). We downloaded the clinical data and transcriptome data of HCC from The Cancer Genome Atlas (TCGA) database, Gene Expression Omnibus (GEO) database, and International Cancer Genome Consortium (ICGC) database. We identified two fibrosis-related molecular subtypes of HCC by consensus unsupervised clustering analysis. Interestingly, these two molecular subtypes significantly differed in overall survival (OS) and clinical characteristics. Besides, the most minor absolute shrinkage and selection operator (Lasso) and multivariate Cox regression analysis were performed to develop a novel prognostic model by three genes (including KPNA2, LPCAT1, and AKR1D1). There was a statistically significant difference in OS between the high-risk and low-risk groups. The area under the ROC curve (AUC) of OS in 1-, 3-, and 5-year were satisfactory. Besides, the risk score was connected with critical clinical characteristics and could be an independent factor in predicting prognosis. Then, the nomogram was built by incorporating risk scores with clinical parameters. Additionally, the risk score was remarkedly correlated with TME and drug susceptibility. Finally, the results of H&E staining and immunohistochemistry of Ki67 showed that the tumor of higher-risk patients are more malignant. The FRGs-based subtype and signature explain the HCC heterogeneity, which might provide a new method to develop a more efficient treatment.

Recent years have seen an increase in the incidence of acne in adults and teenagers resulting in active research in acne treatment. In the present study, we aimed to improve the anti-acne effects of Hwangryeonhaedok-tang (HHT) through fermentation using a kimchi-derived lactic acid bacterium. Lactiplantibacillus plantarum WiKim0111 (Lpb. plantarum WiKim0111) was selected to ferment HHT due to its strong antioxidant activity and its ability to effectively survive and grow in the HHT medium. The antimicrobial activity of HHT and fermented HHT (FHHT) against Cutibacterium acnes (C. acnes) was determined using an agar well diffusion assay. FHHT exhibited stronger antimicrobial, radical scavenging, and superoxide dismutase activity compared to HHT. RAW 264.7 cells pretreated HHT or FHHT showed significantly lower production of nitric oxide, interleukin (IL) -1β, and IL-6 than lipopolysaccharide -treated cells, with FHHT demonstrating higher efficiency than HHT. Overall, HHT fermented by Lpb. plantarum WiKim0111 displayed enhanced antioxidant and anti-inflammatory properties, as well as greater antibacterial activity against C. acnes compared to unfermented HHT. These findings suggest that FHHT may help mitigate acne symptoms by reducing oxidative stress, modulating inflammation, and directly inhibiting the growth of C. acnes. Further in vitro studies focusing on sebum-producing skin cells, as well as clinical trials evaluating the topical or oral application of FHHT, would be valuable to confirm its efficacy and safety in acne treatment.

To analyze the effects of high body energy reserve (BER) within the oviductal environment and its composition, Nellore cows were fed two different nutritional plans to obtain animals with moderate BER (MBER) and high BER (HBER). After obtaining the groups with different BERs, all animals were subjected to oestrus synchronization and artificial insemination, and 120 hours after ovulation induction, the cows were slaughtered, the reproductive tract was removed, and the ipsilateral oviduct to the corpus luteum was collected and dissected. Analyses were performed only for animals that had an 8-cell embryo in the isthmus. After embryo identification, we evaluated the molecular profiles of extracellular vesicles from oviductal flushing (OF-EVs) and luminal epithelial cells (OV-Cell) and performed histomorphological analysis of oviductal tissue from the ampullary and isthmic oviductal regions. The HBER group presented higher concentrations of ampullary extracellular vesicles (AMP-EVs) and larger sizers of isthmic extracellular vesicles (IST-EVs). The miRNA profile of AMP-EVs showed that the differentially expressed miRNAs were predicted to regulate pathways associated with cell growth, migration, differentiation and metabolism, with the HBER group being more susceptible to insulin modulation. The MBER animals showed greater ampullary vascularization than the HBER animals did. Additionally, the miRNA profile and differential gene expression (DEG) data obtained for ampullary (AMP-Cell) and isthmic (IST-Cell) luminal epithelial cells revealed pathways related to insulin metabolism. Thus, elevated BER may lead to oviductal insulin resistance, affecting normal functioning and, probably, embryo metabolism during early development, thus impacting gestational rates in these animals.

The biologically active peptides derived from broad bean protein, including broad bean albumin 1b (BA1b), have anti-oxidation and anti-glycation effects. However, the potential for BA1b to improve glucose metabolism and the underlying mechanisms remain unexplored. We examined the effects of BA1b on glucose metabolism in diabetic mice and further studied the effects of BA1b on intestinal cells based on small intestinal organoids (SIOs). The results showed that BA1b significantly reduced blood glucose and serum insulin levels, promoted secretion of glucose-dependent insulinotropic polypeptide (GIP), and glucagon-like peptide-1 (GLP-1) in diabetic mice. Similarly, BA1b significantly increased the secretion of GIP and GLP-1 levels in SIOs. Mechanistic studies found that BA1b promoted the secretion of incretin (GIP and GLP-1) by up-regulation of VDAC1 gene expression levels. These findings emphasized the important role of BA1b in improving glucose metabolism and provided a new insight into strategies to explore the effects of functional foods.

Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy, with standard treatment consisting of intensive chemotherapy and corticosteroids. While curative in most cases, this regimen leads to significant toxicity and long-term sequelae. Recent advancements in cancer immunotherapy, including chimeric antigen receptor T cells and bispecific T cell engagers, have improved outcomes, yet are limited by toxicity and immune escape by target downregulation. Thus, novel less toxic treatment modalities are highly warranted. The tumor mutational burden in ALL is low, which results in a low number of potentially immunogenic neo-antigens that could be used as targets for neo-epitope-specific therapeutic cancer vaccines. However, recent findings in solid cancer demonstrate that it is not the quantity but the quality of neo-antigens in the tumor that determine the tumor-specific immune response. Furthermore, novel sequencing techniques such as long-read sequencing and optical genome mapping can identify unknown genetic aberrations that may be targeted by neo-antigen vaccines. In ALL, both the ETV6-RUNX1 and BCR-ABL1 fusion genes, and the RAS-isoform mutations are frequent, and these genomic alterations generate immunogenic neo-epitopes. Additionally, therapeutic cancer vaccinations are well suited for ALL as the tumor burden is extremely low at time of a potential post-induction vaccination therapy, and patients are relatively young and are therefore less affected by immunosenescence. Thus, we envisage that neo-antigen specific therapeutic cancer vaccines could pose an important modality in future treatment algorithms for ALL.

G-protein-coupled receptors are membrane-bound proteins that control physiological and psychological activities such as hormonal regulation, sensory signaling and neurotransmission in the human body. The photoisomerization capability of azobenzenes has enabled the development of a new generation of photoswitchable drugs that offer greater efficacy while reducing the side effects typical of conventional treatments. Serotonin binding to the serotonin receptor can be controlled by conjugating it with <i>cis</i> and <i>trans</i> photoswitchable isomers of azobenzene at the hydroxyl position. We have used a combination of molecular dynamics (MD) simulation and free-energy methods for receptor binding to <i>trans</i>-azobenzene-fused-serotonin (TAS) and <i>cis</i>-azobenzene-fused serotonin (CAS) ligand systems. The CAS ligand experienced greater conformational fluctuations in comparison to the TAS ligand. Binding free-energy values showed significant differences between the CAS-receptor and TAS-receptor complexes, indicating that the TAS ligand binds actively to the serotonin receptor as compared to the CAS ligand. It also revealed that the presence of a lipid membrane has a strong influence on the stability and dynamics of the receptor-ligand complex and, thus, on the differential binding free-energy values. The change in binding free energy between the CAS-receptor and TAS-receptor complexes is found to be entropically driven. A qualitative and quantitative analysis of the stacking interactions between aromatic residues of interest and each ligand indicates that T-type stacking interactions predominate and particularly Trp327 and Phe330 residues contribute notably to the stability of the TAS-receptor complex. The study indicates that by leveraging the photoswitchable properties of azobenzenes, a photoswitchable serotonin-based drug with tunable binding affinity can be designed, which may facilitate advanced drug discovery and therapeutic approaches for mental health treatments.

The cell cycle is a fundamental process in eukaryotic biology and is accordingly controlled by a highly conserved core signaling cascade. However, whether recently evolved proteins also influence this process is unclear. Here, we systematically map the influence of evolutionarily recent transcription factors (TFs) on human cell cycle progression. We find that the genomic targets of select young TFs, many of which belong to the rapidly evolving Krüppel-associated box zinc-finger protein (KZFP) family, exhibit synchronized cell cycle expression. Systematic perturbation studies reveal that silencing recent TFs disrupts normal cell cycle progression, which we experimentally confirm for ZNF519, a simian-restricted KZFP. Furthermore, we show that the therian-specific KZFP ZNF274 sets the cell cycle expression and replication timing of hundreds of clustered genes, many of which are KZFPs. These findings highlight an underappreciated level of lineage specificity in cell cycle regulation.

Huntington's disease (HD) is a progressive, inherited neurodegenerative disorder characterised by motor, cognitive, and neuropsychiatric dysfunction for which several mouse models have been developed. Knock-in models, such as zQ175, retain the genetic context observed in people with HD by introducing CAG repeats into the native huntingtin gene. In this study, we conducted a comprehensive, longitudinal analysis of phenotypic changes in the zQ175 mouse, with a focus on exploring the emergence of complex cognitive processes. Our findings indicate that robust cognitive and motivational deficits precede motor dysfunction in this model, with some apparent sex differences. Specifically, male zQ175 mice were slower to habituate to a novel environment and they showed impaired sensorimotor gating, in comparison to female mice. By 12 weeks old, cognitive deficits were observed in zQ175 mice of both sexes on a Pavlovian classical conditioning task. Reduced motivation to work for reward was identified as early as 27 weeks, while attentional and visuospatial deficits were also detected in the 5-choice serial reaction time task. Implicit learning deficits were identified at 30 weeks. zQ175 mice were hypoactive at ∼24 weeks but became hyperactive by 60 weeks of age. Motor impairments emerged by 24 weeks for females and 48 weeks for males. Thus, we observed a wide range of cognitive deficits (attentional, visuospatial, sensorimotor, instrumental and implicit learning), as well as a gradual progression of motor changes. This detailed phenotypic timeline establishes the face validity of this model insofar as these mice present with complex neuropsychiatric and cognitive impairments that are evident in people with HD.

No abstract available.

Anemia is a hallmark of myelodysplastic syndromes/neoplasms (MDS) and most patients with MDS chronically require red blood cell transfusions. Due to the body's inability to excrete excess iron, patients are at increased risk of iron overload, often defined by ferritin levels >1000 ng/mL. Iron overload can cause progressive organ damage from iron deposition in tissues and has been linked to increased mortality. In MDS patients undergoing allogeneic hematopoietic cell transplantation (HCT), iron overload has also been associated with increased non-relapse mortality, decreased overall survival, and a higher incidence of relapse. Prospective and retrospective studies have demonstrated the safety and clinical benefit of iron chelation therapy (ICT) in lower-risk MDS. Despite some common adverse effects associated with ICT, such as renal toxicity and gastro-intestinal symptoms, managing iron levels remains essential in transfusion-dependent MDS patients, and those who are undergoing HCT to optimize pre-transplant conditions, and enhance post-transplant outcomes.

Exercise-induced fatigue refers to a temporary decline in physiological function resulting from prolonged or high-intensity exercise, which is characterized by decreased muscle strength, diminished exercise performance, and heightened subjective feelings of fatigue. The study of exercise fatigue holds significant importance not only in competitive sports and public health, but also extends to medicine, military applications, and occupational safety. MicroRNA (miRNA) represents a class of non-coding RNA that plays a pivotal role in the regulation of gene expression. The involvement of miRNAs in exercise-induced fatigue has garnered increasing attention within the scientific community. This article provides an overview of fundamental concepts and biological functions associated with miRNAs, defines and classifies exercise fatigue while outlining its physiological changes, emphasizes alterations in miRNA expression during episodes of exercise-induced fatigue, and conducts an in-depth analysis regarding the mechanisms through which miRNAs influence this phenomenon via modulation of energy metabolism, inflammatory responses, and oxidative stress. Furthermore, this article anticipates future research directions as well as potential clinical applications for miRNAs concerning exercise-induced fatigue. This review holds significant importance for elucidating the molecular mechanisms underlying exercise-related fatigue while fostering advancements within sports medicine and rehabilitation science.

Novel three-dimensional porous composites of alginate-pectin (A/P) with zinc- or manganese-substituted hydroxyapatites (A/P-ZnHA and A/P-MnHA) were synthesized via lyophilization and calcium cross-linking. Powder X-ray diffraction and infrared spectroscopy analyses confirmed single-phase apatite formation (crystallite sizes < 1 µm), with ZnHA exhibiting lattice contraction (*c*-axis: 6.881 Å vs. 6.893 Å for HA). Mechanical testing revealed tunable properties: pristine A/P sponges exhibited an elastic modulus of 4.7 MPa and a tensile strength of 0.10 MPa, reduced by 30-70% by HA incorporation due to increased porosity (pore sizes: 112 ± 18 µm in the case of MnHA vs. 148 ± 23 µm-ZnHA). Swelling capacity increased 2.3-2.8-fold (125-155% vs. 55% for A/P), governed by polysaccharide interactions. Scanning electron microscopy investigation showed microstructural evolution from layered A/P (<100 µm) to tridimensional architectures with embedded mineral particles. The A/P-MnHA composites demonstrated minimal cytotoxicity for the NCTC cells and good viability of dental pulp stem cells, while A/P-ZnHA caused ≈20% metabolic suppression, attributed to hydrolysis-induced acidification. Antibacterial assays highlighted A/P-MnHA's broad-spectrum efficacy against Gram-positive (<i>Bacillus atrophaeus</i>) and Gram-negative (<i>Pseudomonas protegens</i>) strains, whereas A/P-ZnHA targeted only the Gram-positive strain. The developed composite sponges combine cytocompatibility and antimicrobial activity, potentially advancing osteoplastic materials for bone regeneration and infection control in orthopedic/dental applications.

Different subtypes of breast cancer pose great challenges for precision therapy, especially triple-negative breast cancer (TNBC), because it lacks effective therapeutic targets and is highly resistant to chemotherapy. In this study, the transmembrane protein ITM2A was systematically identified as a novel prognostic biomarker and potential therapeutic target for TNBC. ITM2A was found to be significantly under expressed in TNBC tissues, as revealed by differential expression profiling. Furthermore, patients exhibiting low ITM2A expression demonstrated worse overall survival (OS), recurrence-free survival (RFS), and distant metastasis-free survival (DMFS). A combined multi-omics analysis revealed a significant association between low ITM2A expression and immunosuppressive microenvironmental features. It is noteworthy that the ITM2A high-expression group exhibited substantial clinical benefits in anti-PD-L1 treatment (AUC=0.982) and CAR-T treatment (AUC=0.827). Gene Ontology functional annotation and KEGG pathway enrichment analysis indicated that ITM2A may coordinate anti-tumor immune responses by regulating copper ion metabolic reprogramming and immune checkpoint networks. Pharmacogenomic analysis further confirmed that the expression level of ITM2A was negatively correlated with the sensitivity of etoposide. By establishing the 'immunometabolism-therapeutic response' regulatory axis of ITM2A, this study hopes to provide an innovative theoretical basis for the targeted treatment of TNBC and the precise stratification of immunotherapy.

<h4>Background</h4>Dilated Cardiomyopathy (DCM) is one of the leading causes of non-ischemic cardiomyopathy in the United States (US). The aim of our study is to analyze the general trends in DCM admissions between 2016 and 2021, and analyze social and healthcare disparities in terms of treatments and outcomes.<h4>Methods</h4>National Inpatient Sample (NIS) data for the years 2016 to 2021 were used for the analysis. General population trends were analyzed. Normality of data distribution was tested using the Kolmogorov-Smirnov test and homogeneity was assessed using Levine's test. One-way ANOVA was used after confirmation of normality of distribution to analyze social and healthcare disparities. Subgroup analysis was conducted, with the paired <i>t</i>-test for continuous variables and Fischer's exact <i>t</i>-test for categorical variables to analyze statistical differences. Multivariate regression analysis was conducted to analyze the association of factors that were significant in the one-way ANOVA and paired t/chi square tests. A two-tailed <i>p</i>-value < 0.05 was used to determine statistical significance.<h4>Results</h4>A total of 5262 admissions for DCM were observed between 2016 and 2021. A general declining trend was observed in the total number of DCM admissions, with a 33.51% decrease in total admissions in 2021 compared to 2016. All-cause in-hospital mortality remained stable across the years (between 3.5% and 4.5%). A total of 15.3% of admissions had CRT/ICD devices in place. A total of 425 patients (8.07%) for DCM underwent HT, and 214 admissions for DCM (4.06%) underwent LVAD placements between 2016 and 2021 In terms of interventions for DCM, namely Cardiac Resynchronization Therapy (CRT), Left Ventricular Assist Devices (LVADs) and Heart Transplantations (HTs), significant variance was observed in the mean age of the admissions with admissions over the mean age of 55 had lower number of interventions. Significant variance in terms of sex was observed for DCM admissions receiving HT, with lower rates observed for females. In terms of quarterly income, patients belonging to the lowest fourth quartile had higher rates of LVAD and HT compared to general DCM admissions. In the multivariate regression analysis, age at admission had significant association with lower chances of receiving LVADs and HT among DCM admissions, and significant association with higher chances of all-cause mortality during the hospital stay.<h4>Conclusions</h4>A general declining trend in the total number of DCM admissions was observed between 2016 and 2021. Significant gender disparities were seen with lower rates of females with DCM receiving LVADs and HT. DCM admissions with mean age of 55 and above were found to have significantly lower rates of receiving LVADs and HT, and higher chances of all-cause mortality during the admission.

The human gastrointestinal tract harbors a complex and diverse microbial community. Emerging evidence has revealed bidirectional communication between the gut microbiome and the central nervous system, termed the "microbiota-gut-brain axis". This axis serves as a critical regulator of glial cell function, positioning it as an essential target for ameliorating the onset and progression of ischemic stroke. In this review, we discuss the developments in the relationship between ischemic stroke and neuroinflammation via MGBA. The gut microbiome plays a critical role in signaling to microglia, astrocytes, and other immune components within this axis. We also summarize the interactions between the gut microbiota and glial cells under both healthy and ischemic stroke conditions. Additionally, we also focus on the role of microbiota-derived metabolites and neurotransmitters in ischemic stroke. Furthermore, we investigate the potential of targeting the intestinal and blood-brain barriers to improve MGBA. Finally, we evaluate the preclinical and clinical evidence for dietary interventions, probiotics, prebiotics, and fecal microbiota transplantation in ischemic stroke. A comprehensive understanding of the MGBA is essential for developing MGBA-based treatment for ischemic stroke.

Macrophages are multifunctional immune cells distributed throughout the whole body, and they have functions in antigen presentation, phagocytosis, killing, and immune regulation. As the most widely studied molecule in the netrin family, netrin-1 plays a key role in neuronal navigation, angiogenesis, and cell survival. Macrophage-derived netrin-1 not only regulates neurovascular regeneration through ligand-receptor binding but also influences macrophage phenotypes by modulating polarization, thereby achieving the purpose of promoting or repairing disease damage. In this review, we will summarize the recent research advances on the role of macrophage-derived netrin-1 and its receptors in a variety of inflammatory diseases and cancers.

<h4>Background and objective</h4>Alcohol use accounts for significant morbidity and mortality globally. A wide range of esophageal disorders have been associated with alcohol consumption, most concerning of which is esophageal squamous cell cancer (ESCC). Alcohol use has also been associated with exacerbation of gastroesophageal reflux disease (GERD) symptoms. Despite being one of the first organs to encounter this toxic agent, the molecular effects of alcohol on the human esophagus and the mechanisms by which alcohol exerts its effects on esophageal motility and carcinogenesis remain incompletely defined. The objective of this narrative review is to provide an overview of the effect of alcohol and its toxic metabolite acetaldehyde on cellular signaling, the microbiome, and motility in the esophagus, along with a discussion on the possible mechanisms by which alcohol use increases risk for ESCC.<h4>Methods</h4>A literature search was performed using PubMed, Google Scholar, and Ovid MEDLINE for the period from inception to December 1, 2024. To identify all relevant literature, the following search terms were used: "Alcohol OR ethanol" AND "esophagus OR GERD OR gastroesophageal reflux OR reflux OR cancer OR EoE OR eosinophilic esophagitis OR signaling OR epithelium OR stroma OR fibrosis OR microbiome". Studies published in English with full available text were included.<h4>Key content and findings</h4>This narrative review provides an overview of the effect of alcohol consumption on the human esophagus. The review highlights the dose-dependent risk of ESCC with alcohol use, with increasing risk with higher consumption; with evidence of increased risk even with the smallest amounts of alcohol consistent with the International Agency for Research on Cancer recognition of alcohol as a Group 1 carcinogen. The review also discusses the mechanisms of carcinogenesis, highlighting the role of acetaldehyde, as well as the non-malignant consequences of alcohol on the esophagus, including effects on cellular signaling, the microbiome, and gastro-esophageal reflux disease.<h4>Conclusions</h4>Alcohol consumption continues to exert morbidity and mortality worldwide. Increasing awareness of the multitude of established and potential effects on the human esophagus will allow for rationale supporting at the very least the moderation of alcohol consumption.

Cancer immunotherapy is an innovative treatment approach that leverages the immune system's ability to identify and attack tumor cells. Its goal is to initiate or re-establish the tumor-immune cycle. However, challenges such as limited response rates and adverse immune responses have hindered the further application and advancement of this therapy. Recent progress in nanomedicine, particularly in self-assembled nanomaterials, has attracted significant attention due to their excellent physical and chemical properties. Self-assembled nanoplatforms can be designed to selectively deliver immunoadjuvants, therapeutic drugs, photosensitizers, and sonosensitizers, overcoming the limitations of traditional monotherapies. By utilizing these self-assembled nanoplatforms to synergistically combine cancer immunotherapy with photodynamic therapy (PDT), photothermal therapy (PTT), radiotherapy (RT), and sonodynamic therapy (SDT), it becomes possible to amplify and enhance the immune responses elicited by these localized treatments, thus offering new strategies for cancer therapy. In this review, we discussed various immunotherapy platforms based on the self-assembly of nucleic acids, peptides and proteins, metals, and supramolecules. We also highlight the significant research advancements over the past three years in the use of self-assembled nanomaterials for combination therapies centered on immunotherapy, aiming to provide valuable insights and references for ongoing tumor therapy research.

<title>Abstract</title> <p>Recent studies underscore herpesvirus-associated risks, especially HHV6 and Epstein–Barr virus (EBV), in CAR-T and tumor-infiltrating lymphocyte (TIL) therapies, partly due to ex vivo cell manipulation. EBV-driven B cell outgrowth can occur during TIL expansion, and EBV-transformed B cells may exert immunomodulatory functions, potentially impacting TIL therapy efficacy — areas that remain underexplored. Here, we characterized a spontaneously arising wild-type EBV-transformed B cell line, lcl_burn0214, derived from rectal cancer tumor tissue during ex vivo culture. The lcl_burn0214 cell line has undergone 70 passages and was confirmed as a monoclonal B cell line of patient origin via HLA genotyping, immunophenotyping (CD19, CD20), B cell receptor clonotype analysis. RNA-seq analysis comparing EBV-positive and -negative B lymphoma and lymphoblastoid cell lines revealed that lcl_burn0214’s gene expression closely resembles that of lymphoblastoid cells rather than malignant B lymphomas. In immunocompromised mice, lcl_burn0214 exhibited limited tumorigenicity. The cell line expressed immune checkpoint genes (CD274, CSF1, TNFSF4). Nevertheless, autologous TILs were negative for B cells and exhibited specific anti-EBV activity, as demonstrated by the interferon-gamma response in the ELISPOT assay during coculture with lcl_burn0214. Our findings underscore the importance of monitoring TIL products for EBV-positive B cell contamination during manufacturing to ensure safety and therapeutic efficacy.</p>

<h4>Background</h4>Individuals with thalassemia or sickle cell disease (SCD) are at an elevated risk of developing cancer, yet the full extent of this risk remains inadequately characterised.<h4>Methods</h4>We conducted a systematic review using three databases (MEDLINE, EMBASE and Scopus) to compare the cancer risk in patients with thalassemia and SCD with that of the general population. Secondly, we assessed the impact of other putative risk factors in patients with thalassemia and SCD on cancer risk.<h4>Results</h4>Five studies from four countries were included, encompassing a total of 24,439 individuals overall. The risk estimates of developing cancer were highly variable, being affected by study design, population and comparators recruited, and type of cancer. The results showed an elevated cancer risk in SCD and thalassemia patient groups, with the highest incidence observed for hematologic malignancies (especially myeloid leukaemia in SCD patients). Patients with transfusion-dependent thalassemia had a significantly higher overall cancer risk, particularly hepatocellular carcinoma, compared to both non-transfusion-dependent patients and the general population. Other key factors seem to contribute to the increased cancer risk besides transfusion dependency, viral infections, and use of iron chelators. However, people with hemoglobinopathies appeared to be at lower risk of breast and prostate cancer compared to the general population.<h4>Discussion</h4>The findings are limited by variability among studies, including differences in control groups, inconsistent reporting of outcomes, and missing assessments of confounding factors. Therefore, future research with robust methods is essential to improve cancer surveillance and develop targeted prevention strategies for high-risk populations.

<h4>Background</h4>Genome-wide association studies have provided profound insights into the genetic aetiology of metabolic syndrome (MetS). However, there is a lack of machine-learning (ML)-based predictive models to assess individual genetic susceptibility to MetS. This study utilized single-nucleotide polymorphisms (SNPs) as variables and employed ML-based genetic risk score (GRS) models to predict the occurrence of MetS, bringing it closer to clinical application.<h4>Methods</h4>Feature selection was performed using Least Absolute Shrinkage and Selection Operator. Six ML algorithms were employed to construct GRS models. A fivefold cross-validation was utilized to aid in the internal validation of models. The receiver operating characteristic (ROC) curve was used to select the better-performing GRS model. The SHapley Additive exPlanations (SHAP) was then applied to interpret the model. After extracting GRS, stratified analysis of BMI, age and gender was performed. Finally, these conventional risk factors and GRS were integrated through multivariate logistic regression to establish a combined model.<h4>Results</h4>A total of 17 SNPs were selected for analysis. Among the GRS models, the extreme gradient boosting (XGBoost) model demonstrated superior discriminative performance (AUC = 0.837). The XGBoost's optimal robustness was also validated through five-fold cross-validation (mean ROC-AUC = 0.706). The XGBoost-based SHAP algorithm not only elucidated the global effects of 17 SNPs across all samples, but also described the interaction between SNPs, providing a visual representation of how SNPs impact the prediction of MetS in an individual. There was a strong correlation between GRS and MetS risk, particularly observed among young individuals, males and overweight individuals. Furthermore, the model combining conventional risk factors and GRS exhibited excellent discriminative performance (AUC = 0.962) and outstanding robustness (mean ROC-AUC = 0.959).<h4>Conclusion</h4>This study established a reliable XGBoost-based GRS model and a GRS prediction platform (https://metabolicsyndromeapps.shinyapps.io/geneticriskscore/) to assess individual genetic susceptibility to MetS. This model has high interpretability and can provide personalized reference for determining the necessity of primary prevention measures for MetS. Additionally, there may be interactions between traditional risk factors and GRS, and the integration of both in a comprehensive model is useful in the prediction of MetS occurrence.

One of the most limiting factors in cancer treatment is the difficulty of delivering anticancer agents effectively to tumor sites. To overcome this challenge, this research focuses on developing a carrier that has been modified to possess amphiphilic properties while remaining biodegradable and biocompatible. Amphiphilic esters derived from poly(γ-glutamic acid) by modifying the carboxylate side groups with 4-phenyl-butyl bromide were prepared. These copolymers self-assembled into nanoparticles via nanoprecipitation. The cytocompatibility of the nanoparticles was assessed through lactate dehydrogenase release and metabolic activity of U-87 glioma cells. Fluorescein isothiocyanate labeling demonstrated effective cellular uptake of nanoparticles. These nanoparticles were further decorated with polyethylene glycol (PEG) and a PEG-folic acid conjugate (FA-PEG-NH₂), their sizes being 174 and 156 nm, respectively. Successful grafting was confirmed through ¹H-NMR and FTIR spectroscopy. The nanoparticles were loaded with doxorubicin, and release studies showed their sensitivity to the pH of the environment, the encapsulated drug being released faster at pH 4.2 compared to pH 7.4. Encapsulated doxorubicin's effect on U-87 cells was tested at various concentrations and time points, showing significantly better performance compared to free doxorubicin. These results suggest that those poly(γ-glutamic acid) derivatives hold great promise for improving the delivery of hydrophobic drugs and enhancing cancer treatment.

<h4>Backgroud</h4>Multiple studies have reported that microRNA-103 is unregulated in a variety of tumours, involved in tumorigenesis, and associated with tumour prognosis, so a systematic review and meta-analysis were performed to determine the relationship between microRNA-103 and the prognosis of solid tumours.<h4>Methods</h4>The PubMed, Web of Science, and EMBASE databases were searched to retrieve articles to determine the relationship between microRNA-103 and tumour prognosis. Relevant articles were graded according to the Newcastle-Ottawa Scale (NOS). The 95% confidence interval (CI) was calculated by the fixed-effect/random-effect models and the risk ratio (RR) were summarised.<h4>Results</h4>Eight out of 162 retrieved articles were included in this review, with an average NOS score of 7.2 points. Four studies of tissue samples and four studies of serum samples suggested that the overexpression of microRNA-103 was associated with overall survival (RR = 2.65, 95% CI: 1.79-3.93, P = 0.000 and RR = 3.31, 95% CI: 2.04-5.36, P = 0.000, respectively).<h4>Conclusion</h4>This meta-analysis, combining 9 studies, found that overexpression of miRNA-103 is associated with poor prognosis in solid tumours, particularly in serum samples. Sensitivity analysis confirmed that high tissue expression correlates with poor outcomes. miRNA-103's role in tumor progression suggests its potential as a prognostic biomarker for solid tumors, warranting further research for clinical applications.

<h4>Background</h4>Chronic intestinal hypoxia and accompanying mucosal inflammation is a hallmark of ulcerative colitis. Hyperbaric oxygen therapy involves breathing 100% oxygen under increased atmospheric pressure to increase tissue oxygenation. It reduces systemic and local inflammation and up-regulates hypoxia response pathways, making it an attractive therapeutic option. In this trial we aim to confirm the treatment benefits of hyperbaric oxygen therapy for hospitalized ulcerative colitis patients and assess the long-term durability of treatment effect.<h4>Methods</h4>This prospective, double-masked, multicenter, 1:1 randomized, sham-controlled trial will enroll 126 participants with known or newly diagnosed ulcerative colitis hospitalized for an acute moderate to severe flare. Participants will be randomized to either hyperbaric oxygen therapy with steroids or sham air with steroids. The trial will involve a 5-day intervention period followed by a 12-month observational period with a 90-day standard of care visit and 12-month telephone visit. The primary outcome measure is clinical response defined as complete resolution of rectal bleeding and improvement in stool frequency, without need for in-hospital biologics, small molecules, or colectomy by study day 5. Secondary endpoints include additional key patient-reported outcomes and histo-endoscopic measures of disease activity.<h4>Discussion</h4>Novel and effective treatments are needed for this population to optimize disease outcomes while minimizing treatment-related risks. Demonstrating the ability of hyperbaric oxygen therapy to improve clinical response to steroids and avoid in-hospital rescue therapy has the potential to change the management of hospitalized ulcerative colitis flares.<h4>Trial registration</h4>ClinicalTrials.gov NCT05987852. Registered on August 14, 2023.

The neocortex, which is the site of higher-order cognitive functioning, is comprised of two main neuronal types: excitatory (E) and inhibitory (I). Neurodevelopmental disorders that disrupt the balance of E:I neurotransmission predispose individuals to atypical brain function, highlighting the importance of generating the correct numbers of each neuronal type. During development, neurons with E and I neurotransmission profiles are primarily generated from neural stem and progenitor cells (NPCs), located in the dorsal and ventral telencephalon, respectively. To ensure that correct numbers of each neuronal type are generated, NPC differentiation dynamics vary depending on positional and temporal information and host species. Despite variations in NPC differentiation kinetics and outcomes, proneural genes encoding basic helix-loop-helix (bHLH) transcription factors (TFs) have remained constant as the core drivers of neurogenesis and neuronal subtype specification from fly to human. This high degree of functional conservation raises the question of how proneural TF activity is regulated to control precise neurogenic patterns. In the neocortex, the proneural genes neurogenin 1 (Neurog1) and Neurog2 specify an excitatory neuronal identity in dorsal telencephalic NPCs, whereas achaete-scute family bHLH transcription factor 1 (Ascl1) specifies an inhibitory neurotransmission fate in ventral NPCs, generating interneurons that then migrate tangentially to enter the neocortex. Here, we review our current knowledge of how Neurog1/Neurog2 and Ascl1 functions are regulated to ensure that E:I balance is ultimately achieved in the lissencephalic murine cortex and in gyrencephalic species. Together, these studies point to emergent and conserved features of proneural gene regulation and function across evolutionary time.

Neurodegenerative diseases involve toxic protein aggregation. Recent evidence suggests that biomolecular phase separation, a process in which proteins and nucleic acids form dynamic, liquid-like condensates, plays a key role in this aggregation. Optogenetics, originally developed to control neuronal activity with light, has emerged as a powerful tool to investigate phase separation in living systems. This is achieved by fusing disease-associated proteins to light-sensitive oligomerization domains, enabling researchers to induce or reverse condensate formation with precise spatial and temporal control. This review highlights how optogenetic systems such as OptoDroplet are being used to dissect the mechanisms of neurodegenerative disease. We examine how these tools have been applied in models of neurodegenerative diseases, such as amyotrophic lateral sclerosis, Alzheimer's, Parkinson's, and Huntington's disease. These studies implicate small oligomeric aggregates as key drivers of toxicity and highlight new opportunities for therapeutic screening. Finally, we discuss advances in light-controlled dissolution of condensates and future directions for applying optogenetics to combat neurodegeneration. By enabling precise, dynamic control of protein phase behavior in living systems, optogenetic approaches provide a powerful framework for elucidating disease mechanisms and informing the development of targeted therapies.

Phosphoglucomutase 3 (PGM3) deficiency (OMIM (Online Mendelian Inheritance in Man) #615816) is a rare autosomal recessive congenital disorder of glycosylation that disrupts multiple glycosylation pathways, with few cases reported in the literature. It leads to a broad clinical spectrum ranging from hyper-IgE syndrome (HIES)-like features to severe combined immunodeficiency (SCID). We report a case of a 17-year-old female of Brazilian origin, referred to our center in Portugal for investigation of persistent neutropenia. Her medical history included recurrent infections in early childhood, severe eczema, and autism spectrum disorder. She exhibited persistent neutropenia and T-cell lymphopenia, with elevated IgE levels. Genetic analysis using a next-generation sequencing panel for primary immunodeficiencies identified compound heterozygous likely pathogenic variants in PGM3: a missense variant (c.1475C>T, p.(Thr492Ile)) and a complete gene deletion in the other allele, confirming the diagnosis of PGM3 deficiency. Chronic neutropenia was the main finding that prompted the genetic investigation. Although it is not a defining feature of PGM3 deficiency, it has been reported in nearly half of the cases. In this patient, the clinical presentation has been comparatively milder than the severe phenotypes described in the literature, which highlights the phenotypic variability of this condition and the need for clinical suspicion, even when classical features are absent. The genetic diagnosis has important implications for clinical follow-up and enables appropriate genetic counseling. This case illustrates the clinical variability of PGM3 deficiency and reinforces the role of genetic testing in clarifying the diagnosis, guiding management, and informing long-term follow-up in rare inborn errors of immunity.

Natural antioxidants isolated from fruits, vegetables, herbs and spices have drawn great attention owing to their numerous health-promoting effects. Cinnamaldehyde (CA), an abundant antioxidant in cinnamon spice, has been explored more intensely over the last decade as it has been demonstrated to be effective and safe in the treatment of various diseases. Structurally, a substituted aldehyde group with an unsaturated carbon-carbon double bond with two electrophilic sites for reaction with receptors and enzymes can exert diverse biological effects. Although cinnamon has been traditionally used as a spice and herbal remedy, many studies investigating the most dominant functional compound, CA, and its biological activities have been reported in recent years. This review article intends to present an overview of recent advances in analytical methods and the application of cinnamon extract/oil, CA and its derivatives, CA-polymer/biomolecule conjugates and CA micro/nanosystems in alleviating various chronic diseases including cancer, diabetes, obesity, cardiovascular disease, neurological disorders, osteoarthritis and osteoporosis. Both in vitro and in vivo studies have demonstrated the improved pharmacological efficiency of CA and its derivatives as well as their polymer/drug/biomolecule conjugates and micro/nanoencapsulated forms, suggesting a possible alternative natural therapy and adjuvant therapy with conventional drugs via a synergistic process.

Parkinson’s disease (PD) is characterized by the progressive loss of midbrain dopaminergic (mDA) neurons, leading to severe debilitating motor impairment. Recent single-cell analyses have revealed that specific mDA subpopulations, such as SOX6⁺ AGTR1⁺ neurons, are most vulnerable to degeneration. Current human pluripotent stem cell differentiation protocols fail to selectively generate these subtypes. Here, we describe a multiomic-guided strategy that enriches SOX6 + mDA neurons by combining enhancing Sonic Hedgehog agonism with prolonged Wnt activation. This approach accelerates floor plate specification, increases expression of mDA developmental markers, and substantially increases the yield of SOX6⁺ mature mDA neurons compared to prior attempts. Following intrastriatal transplantation into hemiparkinsonian mice, these cells restored motor function in approximately 4 months and generated grafts containing SOX6⁺ A9-like neurons. Our work thus establishes a reproducible differentiation platform for generating the PD-susceptible mDA subtype, providing a foundation for precision disease modelling and subtype-targeted cell replacement therapies.

Ferroptosis is an iron-dependent form of cell death that contributes to the pathophysiology of chronic obstructive pulmonary disease (COPD). Ferroptosis-associated factors, including acyl-CoA synthetase long-chain family 4 (ACSL4), soluble transferrin receptor 1 (sTfR1), glutathione peroxidase 4 (GPX4), and apoptosis-inducing factor 2, (AIFM2) mediate intracellular iron metabolism, oxidative stress, and lipid peroxidation. Despite their potential clinical relevance, no studies have measured serum levels of these factors with respect to the manifestations of COPD. The study enrolled 179 stable, non-anemic outpatients diagnosed with COPD and 57 age- and sex-matched smokers who did not carry this diagnosis. Clinical characteristics were assessed together with baseline serum levels of the four ferroptosis-associated factors. Moderate and severe exacerbations of COPD were monitored over the following 12 months. Soluble TfR1 levels were higher and GPX4 levels were lower among those in the COPD group compared to smokers without COPD (p = 0.004 and p = 0.002, respectively). The sTfR1/GPX4 ratio was also higher among those in the COPD group (p = 0.001). Multivariate analysis identified low serum GPX4 (OR 5.475; p = 0.001), and high sTfR1/GPX4 (OR 4.293; p < 0.001) as independent predictors of poor performance on the six-minute walk distance test. Additionally, high sTfR1 (HR 1.850; p = 0.004), low GPX4 (HR 2.301; p = 0.001), and high sTfR1/GPX4 (HR 2.223; p < 0.001) were associated with increased risk of moderate exacerbation. High sTfR1 (HR 2.970; p = 0.014), low GPX4 (HR 3.753; p = 0.012), and high sTfR1/GPX4 (HR 3.668; p = 0.009) were also independent predictors of severe exacerbation. Serum levels of ferroptosis-associated factors were significantly different in patients diagnosed with COPD compared to smokers who had not been diagnosed with this disorder. Elevated sTfR1, low levels of GPX4, and higher sTfR1/GPX4 were associated with poor clinical outcomes, including reduced exercise capacity and an increased risk of moderate and severe exacerbations. These findings highlight the potential of ferroptosis-associated factors, particularly the calculated sTfR1/GPX4, in predicting COPD progression and the risk of exacerbation in stable, non-anemic outpatients.

RAF protein kinase acts downstream of RAS in the MAPK pathway, and mutations in the RAF family protein BRAF are frequently observed in adult tumours, with MEK inhibitors proving effective in treatment. However, RAF family protein RAF1 fusions are rare, particularly in pediatric soft tissue tumors, and the efficacy of targeted therapies remains uncertain. This study presents a rare case of a MAP4-RAF1 fusion-positive solid tumor in a child, unresponsive to conventional chemotherapy, surgery, second-line chemotherapy, and sorafenib-targeted therapy. Protein structure simulations predicted trametinib to exhibit optimal efficacy among MEK inhibitors in structural modelling, and it then demonstrated clinical effectiveness in this child. This study underscores the importance of molecular interaction simulations in precise drug screening for clinical decision-making and highlights MEK inhibitors' potential as adjunctive therapy for pediatric RAF1 fusion tumors.

Serum lipid levels, which are influenced by both genetic and environmental factors, are key determinants of cardiometabolic health and are influenced by both genetic and environmental factors. Improving our understanding of their underlying biological mechanisms can have important public health and therapeutic implications. Although psychosocial factors, including depression, anxiety, and perceived social support, are associated with serum lipid levels, it is unknown if they modify the effect of genetic loci that influence lipids. We conducted a genome-wide gene-by-psychosocial factor interaction (G×Psy) study in up to 133,157 individuals to evaluate if G×Psy influences serum lipid levels. We conducted a two-stage meta-analysis of G×Psy using both a one-degree of freedom (1df) interaction test and a joint 2df test of the main and interaction effects. In Stage 1, we performed G×Psy analyses on up to 77,413 individuals and promising associations (P < 10^-5) were evaluated in up to 55,744 independent samples in Stage 2. Significant findings (P < 5 × 10^-8) were identified based on meta-analyses of the two stages. There were 10,230 variants from 120 loci significantly associated with serum lipids. We identified novel associations for variants in four loci using the 1df test of interaction, and five additional loci using the 2df joint test that were independent of known lipid loci. Of these 9 loci, 7 could not have been detected without modeling the interaction as there was no evidence of association in a standard GWAS model. The genetic diversity of included samples was key in identifying these novel loci: four of the lead variants displayed very low frequency in European ancestry populations. Functional annotation highlighted promising loci for further experimental follow-up, particularly rs73597733 (MACROD2), rs59808825 (GRAMD1B), and rs11702544 (RRP1B). Notably, one of the genes in identified loci (RRP1B) was found to be a target of the approved drug Atenolol suggesting potential for drug repurposing. Overall, our findings suggest that taking interaction between genetic variants and psychosocial factors into account and including genetically diverse populations can lead to novel discoveries for serum lipids.

Although the intestinal clock (the circadian timing system in the gastrointestinal tract) is known to direct a wide variety of diurnal nutrients and metabolites, its role in the functioning of extra-intestinal tissues such as the brain remains elusive. Here the role of the intestinal clock in shaping cognitive function is investigated. It is found that Bmal1-iKO mice (mice with Bmal1 [Brain and muscle Arnt-like protein 1] specifically knocked out in the intestine, a mouse line deficient in intestinal clock function) show a defect in cognitive memory irrespective of the time-of-day. Bmal1-iKO-associated cognitive decline is attributed to impaired adenosine signaling and compromised long-term potentiation (LTP) in the hippocampus. Adenosine signaling promotes LTP via enhancing BDNF expression and inhibiting synapse loss. Furthermore, the impairment in adenosine signaling is accounted for by the reductions in intestinal absorption of and hippocampal level of adenosine but not by a change in adenosine receptors. Consistently, adenosine supplementation rescues cognitive deficits associated with the malfunction of the intestinal clock. Moreover, BMAL1 regulates the expression of ADK (adenosine kinase, a primary enzyme for adenosine clearance) in the small intestine and thus promotes intestinal adenosine absorption through REV-ERBα which binds directly to Adk P2 promoter to inhibit its transcription. Together, an unsuspected role of the intestinal clock in controlling cognitive memory is identified, highlighting the intestinal clock as a promising target for the management of cognitive disorders.

Necroptosis is a form of programmed cell death that, when dysregulated, is associated with cancer and inflammatory and neurodegenerative diseases. Here, starting from hits identified from a phenotypic high-throughput screen for inhibitors of necroptosis, we synthesized a library of compounds containing a 7-phenylquinoline motif and validated their anti-necroptotic activity in a novel live-cell assay. Based on these data, we designed an optimized photoaffinity probe for target engagement studies and through biochemical and cell-based assays established receptor-interacting kinase 1 (RIPK1) as the cellular target, with inhibition of necroptosis arising from the prevention of RIPK1 autophosphorylation and activation. X-ray crystallography and mass spectrometry revealed that these compounds bind at the hinge region of the active conformation of RIPK1, establishing them as type I kinase inhibitors. In addition, we demonstrated <i>in vitro</i> synergy with type III kinase inhibitors, such as necrostatin-1 and found that lead compounds protected mice against acute inflammation in necroptosis models <i>in vivo</i>. Overall, we present a novel pharmacophore for inhibition of human RIPK1, a key protein involved in necroptosis, and provide a photoaffinity probe to explore RIPK1 target engagement in cells.

Carpet could serve as a potential vehicle for transmitting <i>Clostridioides difficile</i> endospores, a leading cause of healthcare-associated infections. To develop carpet disinfection practices, a validated recovery method and standard efficacy testing method are necessary. The recovery method was optimized to detect <i>C. difficil</i>e endospores on carpet by using different concentrations of Tween-80 and varying stomaching times. Adding 0.2% Tween-80 followed by stomaching for 3 min and sonication increased the recovery rate of <i>C. difficile</i> endospores to >60%. The efficacy of three disinfectants (two hydrogen peroxide-based products, A and B, and one chlorine-based product, C) and steam was tested against <i>C. difficile</i> endospores on two types of nylon carpet (with water-permeable backing and waterproof backing). The results showed that product B was the most effective among the chemical disinfectants, achieving a 5.8 and 4.9 log₁₀ CFU reduction of <i>C. difficile</i> endospores in 30 min on carpets with water-permeable and waterproof backings, respectively. A steam treatment of 120 s resulted in 4.9 and >6.0 log₁₀ CFU reduction on water-permeable and waterproof backings, respectively. On water-permeable backing, 30 s steam treatment followed by application of H₂O₂-based products A and B resulted in a 4.4 and >6.1 log₁₀ CFU reduction of <i>C. difficile</i> endospores, respectively. Overall, a 120 s steam treatment was more effective than any of the three disinfectants tested. Disinfectant efficacy varied by carpet backing type, underscoring the need for guidance in carpet selection for healthcare safety.<h4>Importance</h4><i>Clostridioides difficile</i>, a spore-forming anaerobic bacterium and a leading cause of healthcare-associated infections, can be transmitted from the floor to other surfaces via air movement. Therefore, disinfection of all floors after cleaning, regardless of type, might be necessary to prevent recurrent <i>C. difficile</i> infections among patients. To develop carpet disinfection practices, a validated recovery method and standard efficacy testing method are necessary. In this study, we first optimized the spore recovery method from carpets. Next, our study demonstrated that carpet backing affected the efficacy of chemical disinfectants and steam against <i>C. difficile</i> endospores. Steam was particularly effective on carpets with waterproof backing, while only product B showed strong efficacy on carpets with water-permeable backing. When steam was combined with a chemical disinfectant, the efficacy of both H₂O₂-based products against <i>C. difficile</i> endospores was enhanced. These findings can inform the development of carpet disinfection practices.

Prior analyses suggested that the type II secretion system (T2SS) of <i>Legionella pneumophila</i> secretes ≥47 proteins beyond its 26 known substrates. Upon examination of mutants of wild-type strain 130b that lack those exoproteins most conserved across the <i>Legionella</i> genus, we discovered that protein "06635" majorly promotes <i>L. pneumophila</i> replication within amoebae. Immunoblotting, proteomics, and whole-cell enzyme-linked immunosorbent assay (ELISA) confirmed that 06635 exists in culture supernatants and on the bacterial outer surface and does so in a T2SS-dependent manner. Bioinformatic analyses identified 06635 as a novel member of the carbohydrate esterase family 4, whose members deacetylate bacterial surface polysaccharides, peptidoglycan, chitins, and/or xylans. Given 06635's T2SS-dependent secretion, low-level amino acid similarity to known peptidoglycan deacetylases, and the unaltered lysozyme resistance of a <i>06635</i> mutant, we pursued the hypothesis that 06635 deacetylates a polysaccharide on <i>L. pneumophila</i>'s surface. Supporting this, the <i>06635</i> mutant exhibited increased binding to both wheat germ agglutinin (i.e., more surface <i>N</i>-acetylglucosamine) and antibodies that recognize acetylated lipopolysaccharide (LPS). Nuclear magnetic resonance (NMR) analysis of isolated mutant vs wild-type LPS confirmed that 06635 promotes LPS deacetylation. Thus, we designated 06635 as PdaA, for <u>p</u>olysaccharide <u>d</u>e<u>a</u>cetylase <u>A</u>. Compatible with its altered surface, the <i>pdaA</i> mutant showed greater autoaggregation, increased biofilm formation, and heightened sensitivity to both polymyxin and human serum. Thus, we hypothesize that, following its secretion via the T2SS, PdaA deacetylates LPS, and perhaps other moieties, impacting many significant processes. While defining PdaA, we identified many more putative substrates of the <i>L. pneumophila</i> T2SS, bringing the size of the T2SS output to approximately 120.IMPORTANCE<i>Legionella pneumophila</i> is the principal cause of Legionnaires' disease, an increasingly common form of pneumonia. Although prior work demonstrated that the bacterium utilizes its type II protein secretion system (T2SS) to survive in aquatic environments and to cause lung infection, the full scope and impact of this <i>Legionella</i> secretion system is still relatively underappreciated. By utilizing an expanded proteomic approach and testing newly made mutants in a wide range of assays, we have determined that the <i>L. pneumophila</i> type II secretome encompasses approximately 120 proteins, and among these proteins is a novel polysaccharide deacetylase (PdaA) that modulates the <i>L. pneumophila</i> surface and lipopolysaccharide, impacting intracellular infection, biofilm formation, and resistance to both antibiotics and human serum. Moreover, since T2SSs and homologs of PdaA were found in many other bacteria, our findings should also have implications for understanding other infectious diseases and environmental processes.

This study seeks to understand cardiac medication use in a large cohort of males with Duchenne Muscular Dystrophy (DMD) followed prospectively with focus on current practices and adherence to consensus directed medical therapy (CDMT). DMD patients have been enrolled in the Advanced Cardiac Therapies Improving Outcomes Network (ACTION) Dystrophinopathy Registry since 2021. Cardiac medication use was analyzed at enrollment and most recent follow-up. CDMT was defined as concurrent use of angiotensin-converting-enzyme inhibitor (ACEi)/angiotensin II receptor blocker (ARB) /angiotensin receptor-neprilysin inhibitor (ARNI) plus beta-blocker (BB) plus mineralocorticoid receptor antagonist (MRA). Two hundred sixty-five males with DMD (median age 17.5 (IQR 14.5-21.5) years) were prospectively followed; median follow-up was 11.5 (IQR 6.2-15.6) months. At most recent follow-up, 153 patients (57.7%) had decreased LV systolic function, 67 (25.3% of the cohort) had moderate or severe dysfunction. For patients with moderate or severe dysfunction, CDMT was used for 49/67 (73.1%) at most recent follow-up, similar to 36/51 (70.6%) at enrollment (p = 0.92). Target doses of CDMT were achieved for 27% of males on ACEi/ARB/ARNI, 28% on BB, and 23% on MRA. Initial analysis of a prospective registry of males with DMD showed that ~ 30% of patients with moderate or severe LV dysfunction were not on CDMT at most recent follow-up and the majority did not reach target dosing. Further understanding regarding the optimal combination of cardiac medications for DMD cardiomyopathy is needed, as is a better understanding of the barriers to CDMT optimization given increasing cardiac causes of death for DMD patients.

No abstract available.

Immunotherapy represents a pivotal therapeutic modality in endometrial cancer (EC). Nevertheless, the efficacy of this treatment is limited to a subset of patients. The present investigation endeavors to amalgamate multi-omics data in order to elucidate the determinants impacting individual immune responsiveness and enhance the optimization of immunotherapy for EC. To differentiate EC patients into non-response (NR) and response (R), multi-omics data from publicly available databases were employed in conjunction with the TIDE computational framework. The validity of these findings was further confirmed through the utilization of the EaSIeR and ImmunoPhenoScore algorithms. The study employed functional enrichment and gene set variant analysis to discern noteworthy disparities in biological pathways across various groups. Moreover, three deconvolution algorithms (ESTIMATE, TIMER, and EPIC) were employed to quantify the tumor microenvironment (TME). Somatic mutation and copy number variant (CNV) analyses were also conducted to identify genomic alterations that impact immunotherapy. Integrated bulk and single-cell RNA sequencing (scRNA-seq) data were employed to identify cell populations linked to efficacy and deduce cell-cell interactions. The immunotherapy response rates were found to be greater in elderly EC patients aged 65 years and above. The NR group of patients displayed notable enrichment in cellular differentiation, angiogenesis, and tumor proliferation characteristics, as evidenced by higher tumor purity and lower expression of immune checkpoints. Conversely, the R group exhibited a stronger correlation with immunity, as indicated by pathway enrichment and composition of TME. Patients in the NR group demonstrated higher frequencies of somatic mutations, with a 2- to 6-fold disparity between the groups in genes such as RPRD1B and CTNNB1. Patients in the R group exhibited elevated mutation scores and higher mutation frequencies at the same mutation loci compared to those in the NR group. Moreover, the incidence of mutations was more prevalent among patients in the R group. In independent cohorts, the Scissor algorithm suggests that macrophages may exert a substantial impact on immunotherapy response in patients with EC. Subsequent analysis unveiled an enrichment of M2-like tumor-associated macrophages (TAMs) within the TME of patients in the NR group. These macrophages facilitate angiogenesis and cell proliferation through intercellular communication with subpopulations such as endothelial and epithelial cells. TME of patients in the R group exhibited an enrichment of M1-like TAMs, which primarily engaged with immune cells via diverse immune-activating factors. Furthermore, immunohistochemistry and flow cytometry demonstrated that responders to immunotherapy had significantly increased infiltration of M1-like TAMs. M1-like TAMs were shown to inhibit proliferation and migration of Ishikawa cells in co-culture assays. This research offers a comprehensive insight into the multi-omics level factors influencing the immunotherapy response of EC patients, emphasizing the influence of genomic variants and TAMs on said response. This contributes to an enhanced comprehension of the biological mechanisms underlying EC immunotherapy response and aids in the optimization of EC immunotherapy.

Kudzu's invasive nature has contributed to its classification as a weed, as it frequently outcompetes native plant species, leading to extensive overgrowth. Efforts to control kudzu have proven challenging, with moderate success using physical or biological agents. In this study, we evaluated the effects of two such control agents, ultraviolet C radiation and <i>Myrothecium verrucaria</i>, to significantly increase the production of tuberosin, a phytoalexin isoflavone. Our findings demonstrate that estrogenic activity of tuberosin is cell-type-dependent, displaying antagonist or competitive inhibition when combined with 17-β-estradiol in the estrogen receptor (ER) positive cell lines MCF-7 and T-47D, while showing dose-dependent agonist activity in HEK293 cells transfected to express both ER receptors (α and β). Tuberosin was shown to modulate ER pathways, alter ER-mediated gene expression, and increase cell proliferation in a dose-dependent manner while maintaining expression of the ERα protein. Binding affinity and docking simulations confirmed tuberosin binding to the ERα pocket in a similar but weaker manner compared to synthetic estrogen. Tuberosin-treated endothelial cells suppressed vascular network assembly and maturation without affecting the cellular proliferative capacity. The presented studies leverage current kudzu management methods to naturally produce tuberosin, examine cell-type-specific effects, and support further investigation as an antiestrogen for breast cancer treatment.

<h4>Introduction</h4>Research findings show a substantial correlation between Crohn's disease and psoriasis. However, the exact cause or pathogenesis of the concurrent manifestations of these two conditions in the same individuals remains uncertain. This research aimed to scrutinize the important molecules and mechanisms responsible for the concomitance of Crohn's disease and Psoriasis by using quantitative bioinformatics utilizing a publicly available RNA sequencing repository.<h4>Methods</h4>The database Gene Expression Omnibus were assessed, specifically for Crohn's disease (GSE95095) and psoriasis (GSE13355). The 'limma' library of the R programming syntax is employed to identify differentially expressed genes. The Search Tool for Interacting Genes dataset was utilized to study the interaction between proteins networks. The Cytoscape software was utilized to efficiently view and analyse these Protein-Protein Interaction networks. The ctoHubba Cytoscape plugin helps in the selection of hub genes. These hub genes have been confirmed using data from GSE102133 for Crohn's disease and GSE14905 for psoriasis. The ROC curves were utilized in this study to assess the diagnostic value of the hub genes. Moreover, new research involving gene-set enriched studies and the study of immunological surveillance associated with these specific genes is attainable.<h4>Results</h4>Among the identified common DEGs, 40 genes were downregulated and 37 were upregulated, totaling 77 genes. Crohn's disease and Psoriasis had a higher concentration of pathways associated with inflammation. After validation, functionality of hub genes was confirmed for S100A12, CXCL8, IL1RN, S100A9, CXCL10, MMP1, CXCL1, FPR1, CXCR2, and S100A8. The hub genes showed an increase in expression in response to neutrophil infiltration. The expression of S100A12, CXCL8, IL1RN, S100A9, CXCL10, MMP1, CXCL1, FPR1, CXCR2, and S100A8 was found to be significantly linked to immune processes such as neutrophil activation, neutrophil chemotaxis, and neutrophil migration associated with Crohn's and Psoriasis disease.<h4>Conclusions</h4>This bioinformatics study has elucidated S100A12, CXCL8, IL1RN, S100A9, CXCL10, MMP1, CXCL1, FPR1, CXCR2, and S100A8 as the central genes in the pathogenesis of CD and Psoriasis comorbidity. The significance of neutrophil infiltration in promoting inflammatory and immune-mediated dysfunction seems to be crucial in the etiology of concurrent Crohn's and Psoriasis, offering an avenue for diagnostic and therapeutic methods.

Identifying effective therapeutic targets for Parkinson's disease (PD) is challenging, with no current disease-modifying therapies available. To address this, The Michael J. Fox Foundation for Parkinson's Research launched the Targets to Therapies (T2T) initiative, uniting experts to prioritize and validate promising targets. T2T aims to develop validation strategies, create comprehensive target data profiles, and build tools to support drug development, ultimately accelerating the discovery of new therapies for PD patients.

Periodontitis is a chronic inflammatory disease initiated by plaque microorganisms, with the regulatory mechanisms of its progression being a primary research focus. Ferroptosis, a unique form of cell death driven by iron-dependent lipid peroxidation, has been increasingly recognized for its crucial role in modulating chronic inflammation. This study focused on the molecular mechanisms by which plaque microorganisms and the inflammatory microenvironment trigger ferroptosis in periodontal cells, elucidating how ferroptosis in these cells promotes periodontitis progression. Additionally, the potential exacerbation of periodontitis through ferroptosis in systemic diseases such as Alzheimer's disease, nonalcoholic steatohepatitis, chronic obstructive pulmonary disease, and type 2 diabetes is discussed. This review aims to provide new theoretical foundations and strategies for the treatment of periodontitis.

Refining the classification of autism spectrum disorder (ASD) subtypes is essential for advancing personalized interventions strategies, given the substantial heterogeneity in phenotypic clinical symptoms among individuals with ASD. Thus, the current study integrated normative modeling, resting-state fMRI data, clinical assessment, and eye-gaze patterns to investigate potential ASD subtypes. By incorporating both static and instant dynamic (strength and variability) functional connectivity as predictive variables within the normative models, we aimed to delineate multi-level functional developmental trajectories. Our comprehensive analysis of 1046 participants (479 with ASD, 567 typical development) identified two distinct neural ASD subtypes with unique functional brain network profiles despite comparable clinical presentations. One ASD subtype was characterized by positive deviations in the occipital network and cerebellar network, coupled with negative deviations in the frontoparietal network, default mode network, and cingulo-opercular network. Conversely, the other subtype exhibited an inverse pattern of functional deviations across these networks. Furthermore, an independent cohort of 21 ASD individuals revealed that these neural subtypes were also associated with distinct gaze patterns assessed by two autism-sensitive eye-tracking tasks focused on preference for social cues. These findings emphasize the complexity and heterogeneity of ASD, uncovering the presence of neurobehavioral subtypes that extend beyond simple neural variations and manifest in diverse functional developmental patterns and behavioral presentations. This study underscores the importance of adopting personalized intervention strategies that cater to the individual needs of each person, rather than relying on a standardized, one-size-fits-all approach, despite the presence of similar clinical symptoms.

<h4>Objective</h4>Attention-deficit hyperactivity disorder (ADHD) is a common neuropsychiatric disorder with a significant genetic component. The latest genome-wide association study (GWAS) meta-analysis of ADHD identified 27 whole-genome significant risk loci in the European population. However, genetic risk factors for ADHD are less well-characterized in the Asian population, especially for low-frequency / rare variants.<h4>Methods</h4>In this study, we aimed to investigate the contributions of both common and low-frequency / rare variants to ADHD in a Hong Kong sample. Our sample comprised 279 cases and 432 controls who underwent genotyping using the Illumina Infinium Global Screening Array. We employed various analytical methods at different levels, while also leveraging multi-omics data and large-scale summary statistics to comprehensively analyze the genetic basis of ADHD.<h4>Results</h4>We identified 41 potential genomic risk loci with a suggestive association (p < 1e^-4), pointing to 111 candidate risk genes, which were enriched for genes differentially expressed during late infancy brain development. Furthermore, tissue enrichment analysis implicated the involvement of the cerebellum. At the polygenic level, we also discovered a strong genetic correlation with resting-state functional MRI connectivity of the cerebellum involved in the attention/central executive and subcortical-cerebellum networks. In addition, an accumulation of ADHD common-variant risks found in European ancestry samples was found to be significantly associated with ADHD in the current study. In low-frequency / rare variant analyses, we discovered the correlations between ADHD and collapsing effects of rare damaging variants in TEP1, MTMR10, DBH, TBCC, and ANO1. Based on biological and functional profiles of the potential risk genes and gene sets, both common and low-frequency / rare variant analyses demonstrated that ADHD genetic risk was associated with immune processes.<h4>Conclusions</h4>These findings re-validate the abnormal development of the neural system in ADHD and extend the existing neuro-dysfunction hypothesis to a multi-system perspective. The current study identified convergent risk factors from common and low-frequency / rare variants, which implicates vulnerability in late-infancy brain development, affecting especially the cerebellum, and the involvement of immune processes.

<h4>Background</h4>Hymenolepis nana (H. nana) is a zoonotic parasitic worm that parasitizes the small intestines of humans and rodents. Ulcerative colitis (UC) is a chronic and recurrent inflammatory bowel disease. Current symptom-based clinical treatments do not alter the natural course of UC, and mucosal healing has become a primary therapeutic goal for UC. However, the regulatory role of excretory/secretory products (ESPs) from H. nana adult worms in repairing the damaged intestinal mucosal barrier remains unclear.<h4>Methods</h4>This study investigated the protective effects of ESPs on intestinal mucosal integrity by using a dextran sulfate sodium (DSS)-induced colitis mouse model and a mouse small intestine organoid inflammation model. Histopathological alterations of mouse intestinal tissues were determined by pathological staining; the alterations in mucins, tight junction proteins, cytokines, and the number of various intestinal cells were detected by Western blotting (WB), immunohistochemistry (IHC), immunofluorescence (IF) and real-time quantitative polymerase chain reaction (RT-qPCR), etc. RESULTS: ESPs significantly improved DSS-induced intestinal damage in mice. Meanwhile, ESPs increased mucins and tight junction proteins expression and promoted intestinal stem cell proliferation and differentiation, thereby maintaining intestinal mucosal barrier integrity and alleviating UC in mice. In the DSS-induced inflamed small intestinal organoid model, ESPs reduced organoid damage and promoted the proliferation and differentiation of intestinal stem cells. The protective mechanism of ESPs might be related to the activation of the tuft/IL-13 signaling pathway, regulating intestinal barrier function and promoting the regeneration of intestinal stem cells.<h4>Conclusions</h4>In conclusion, H. nana-derived ESPs intervention facilitates healing of intestinal mucosa to alleviate UC in mice, enriching the feasibility and selectivity of "helminthic therapy."

<h4>Background</h4>Despite the high prevalence of cerebral ischemic stroke, effective clinical treatments remain limited. With the development of regenerative medicine, induced neural progenitor cells (iNPCs) demonstrate ideal potential and good availability for autologous transplantation therapy. However, current differentiation protocols for iNPCs still have room for improvement in terms of purity, reproducibility, scalability and differentiation potential.<h4>Methods</h4>We aimed to develop a scalable, stable, and efficient 3D aggregate-based method for iNPC production in suspension culture, avoiding detrimental cell dissociation and replating processes. We evaluated the therapeutic potential of iNPCs in the chronic phase of a transient middle cerebral artery occlusion (tMCAO) mouse model and explored iNPC subpopulations via single-cell RNA sequencing to elucidate their pleiotropic therapeutic potentials.<h4>Results</h4>iNPCs generated from three iPSC lines displayed high NPC marker expression and an average 176-fold cell expansion over the 12-day culture period. These iNPCs could spontaneously differentiate into both neurons and glial cells in vitro. In the tMCAO model, transplanted iNPCs remodeled the microenvironment by alleviating neuroinflammation, inhibiting chronic microgliosis and astrogliosis, promoting M2 polarization of microglia, and preserving astrocytic morphology in the ischemic penumbra. Mechanistically, iNPCs can be divided into four subpopulations, with neuroepithelia being the most abundant and capable of rapidly replenishing damaged cells and mitigating microenvironmental deterioration.<h4>Conclusions</h4>We developed a simple and efficient 3D aggregate-based method for iNPC differentiation. These iNPCs showed excellent potential for post-stroke recovery and represent a valuable tool for clinical translation.

Ferroptosis, an iron dependent mode of cell death, is characterized by oxidative damage to cell membrane by uncontrolled lipid peroxidation. Prevention of lipid peroxide formation or its rapid neutralization is critical to avert ferroptosis. Selenoproteins such as glutathione peroxidase 4 (GPX4) and thioredoxin reductase 1 (TXNRD1) are critical enzymes which prevent ferroptosis. Peroxiredoxin 6 (PRDX6) is a lipid peroxide specific peroxidase enzyme with moonlighting functions that affect ferroptosis sensitivity in a multitude of ways. Further, it has recently been discovered that PRDX6 also acts as an intracellular carrier of selenium, allowing efficient utilization of organic and inorganic sources of selenium for synthesis of selenoproteins. Thus, PRDX6 can affect abundance and functionality of anti-ferroptotic selenoproteins GPX4 and TXNRD1. This review is focussed on understanding the different functions of PRDX6 in ferroptosis in light of its newly discovered role in selenoprotein synthesis.

<h4>Background</h4>Malnutrition and epilepsy share a complex bidirectional relationship, with malnutrition serving as a potential risk factor for epilepsy development, while epilepsy, in turn, often exerts profound effects on nutritional status. Nutritional interventions have emerged as a critical adjunctive approach in epilepsy management.<h4>Aim</h4>To explore the multifaceted associations between malnutrition and epilepsy, structured into three primary sections: (1) Elucidating the impact of malnutrition as a risk factor for epilepsy onset; (2) Examining the reciprocal influence of epilepsy on nutritional status, and (3) Evaluating diverse nutritional interventions in the management of epilepsy.<h4>Methods</h4>A systematic search was conducted across PubMed, Scopus, and Web of Science databases utilizing defined keywords related to malnutrition, epilepsy, and nutritional interventions. Inclusion criteria encompassed various study types, including clinical trials, animal models, cohort studies, case reports, meta-analyses, systematic reviews, guidelines, editorials, and review articles. Four hundred sixteen pertinent references were identified, with 198 review articles, 153 research studies, 21 case reports, 24 meta-analyses, 14 systematic reviews, 4 guidelines, and 2 editorials meeting the predefined criteria.<h4>Results</h4>The review revealed the intricate interplay between malnutrition and epilepsy, highlighting malnutrition as a potential risk factor in epilepsy development and elucidating how epilepsy often leads to nutritional deficiencies. Findings underscored the importance of nutritional interventions in managing epilepsy, showing their impact on seizure frequency, neuronal function, and overall brain health.<h4>Conclusion</h4>This systematic review emphasizes the bidirectional relationship between malnutrition and epilepsy while emphasizing the critical role of nutritional management in epilepsy treatment. The multifaceted insights underscore the need for a holistic approach to addressing nutritional aspects alongside conventional epilepsy management strategies.

Inflammatory bowel disease (IBD)-associated colorectal cancer (CRC) is a serious global health issue. Owing to the successful mimicking of the entire process of CRC induced by IBD, azoxymethane (AOM) and dextran sulfate sodium (DSS)-induced colitis-associated cancer (CAC) animal models have been widely used to study the carcinogenic mechanisms of IBD-associated CRC and aid in the discovery of new candidate target drugs for CRC treatment. This article summarizes the molecular characteristics and mechanisms of the AOM/DSS-induced CAC animal model and reports the effects and mechanisms of dietary active ingredients such as phenolics, flavonoids, terpenes, polysaccharides, alkaloids, and other food extracts in the prevention and treatment of AOM/DSS-induced CAC in preclinical trials. Further research and application of dietary active ingredients against AOM/DSS-induced CAC are needed in the future. In conclusion, this study may provide insights into the potential application of active dietary ingredients in the treatment of CAC.

<h4>Introduction</h4>In an increasingly aging society, testing hippocampal-dependent cognition in a quick and low resource manner will be crucial in: assessing the potential benefits of lifestyle choices and interventions affecting cognitive ageing (such as those involving exercise, diet, and sleep); detecting pathological aging, such as in Alzheimer's disease, where hippocampal degeneration occurs relatively early on.<h4>Methods</h4>Over 300 participants aged 18-89 completed three cognitive tests, namely the Addenbrooke's Cognitive Examination-III (ACE-III), The Four Mountains Task (4MT), and a new task introduced here, the Spaces and Sequences Episodic Video Task (SSEVT). Hippocampal tissue is particularly vulnerable to aging, and the 4MT and SSEVT were designed to be hippocampal-dependent. Accordingly, we tested the hypothesis that 4MT and SSEVT performance would be significantly compromised by aging. As an initial proof-of-concept exploration of these tests' ability to detect pathological aging, such as in Alzheimer's disease, we compared 10 patients with Mild Cognitive Impairment (MCI) with matched subsamples of the older group (Healthy ageing, HA).<h4>Results</h4>Supporting the hippocampal-aging related hypothesis, 4MT and SSEVT scores showed appreciably stronger age-related declines than ACE-III scores. The middle-aged group (mean: ∼51 years) were significantly worse than the young group (mean: ∼21 years) on the 4MT (Cohen's d = 0.724) and the SSEVT (Cohen's d = 0.443); and the older group (mean: ∼71 years) were significantly worse than the middle-aged group on the SSEVT (Cohen's d = 0.724). Neither pattern was seen for ACE-III. Suggestively, the MCI patients performed worse than the matched HA group on the 4MT (consistent with previous work), and on our novel SSEVT, but not on the ACE-III.<h4>Discussion</h4>We conclude that the 4MT and SSEVT may be suitable for assessing lifestyle choices and interventions affecting cognitive ageing. We also propose that these findings provide an initial proof-of-concept for these tests' ability to detect pathological aging in its early stages and support further exploration of this with larger clinical samples.

<h4>Objective</h4>To investigate the sex differences in environmental exposure to per- and poly-fluoroalkyl substances (PFAS) in ischemic heart disease (IHD) and to identify potential targets for future prevention and treatment of PFAS-associated IHD.<h4>Methods</h4>The Global Health Data Exchange database was used to explore the sex differences in IHD mortality and morbidity. The National Health and Nutrition Examination Survey (NHANES) database was used to identify sex differences in response to environmental exposure to PFAS, including survival probability and dose-response. The Comparative Toxicogenomics Database and Gene Expression Omnibus databases were used to search for critical signaling pathways involved in IHD pathogenesis and potential targets for the prevention and treatment of PFAS-associated IHD. The binding stability of these complexes was evaluated by molecular docking and molecular dynamics simulations.<h4>Results</h4>Globally, the mortality, morbidity, years of life lost, and years lived with disability are higher for men than women. Among 42,742 participants from NHANES, including IHD and control groups as well as PFAS-affected IHD subjects, men had significantly lower survival rates than women. Four PFAS exposures, including perfluorooctane sulfonamide, perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), and 2-(N-methyl-PFOSA) acetate, significantly worsened the survival of patients with IHD and interacted with 105 human genes associated with cardiovascular diseases. Combining differentially expressed genes from the pluripotent stem cell-derived cardiomyocyte dataset, five promising genes-CASP3, PDK4, GDF15, RPL17, and CTNNB1-were identified as having high binding stability to PFAS.<h4>Conclusion</h4>Men with IHD have significantly worse survival rates than women, yet women are more susceptible to PFOA and PFOS toxicity. This study also identifies several PFAS receptor genes that affect key pathways in IHD pathogenesis, which are promising potential targets for future prevention and treatment of PFAS-associated IHD.

The endoplasmic reticulum (ER) is the most metabolically active organelle in cells, and recent research has shown that abnormal ER function is involved in the occurrence and development of acute kidney injury (AKI), but the underlying molecular mechanism needs to be further elucidated. Here, we review the biological functions of the ER in cellular metabolism, explore the current research progress on the role of the ER in different triggers of AKI, and summarize the ER stress inhibitors discovered thus far. Finally, we explore the possibility of targeting ER homeostasis as a therapeutic target for AKI.

<h4>Background</h4>Gastric cancer (GC) is characterized by heterogeneous tumor microenvironment (TME) with various cell types contributing to disease progression and patient outcomes. This study aims to dissect the single-cell transcriptomic landscape of GC, highlighting the role of tumor-associated macrophages (TAMs) and establishing a novel prognostic signature based on high oxidative phosphorylation (OXPHOS) macrophages.<h4>Methods</h4>Single-cell sequencing data from paired GC and normal stomach tissues, obtained from the GEO database (GSE184198), were processed to reveal cellular heterogeneity and identify TAM subsets with high OXPHOS activity. Using the TCGA STAD dataset, survival analyses were conducted on 435 GC patients to establish a high-OXPHOS-macrophage-related prognostic signature.<h4>Results</h4>We identified eight distinct cell types within the GC TME, indicating significant cellular heterogeneity. Macrophages, particularly TAMs, were found in greater numbers in tumor tissue, with the C3 macrophage subset exhibiting the highest OXPHOS score. A 19-gene high-OXPHOS-macrophage-related prognostic signature was constructed, stratifying patients into different risk categories with significant survival differences (P<0.05). NPC2, LY96, and TPP1 were identified as key macrophage-expressed markers, correlating with prognosis. Cell communication analysis revealed increased interaction in tumor tissues, especially involving NPC2, LY96, and TPP1 positive macrophages, which facilitated tumorigenesis and immune evasion.<h4>Conclusion</h4>The high-OXPHOS-macrophage-related prognostic signature derived from scRNA-seq data provides valuable insights into GC patient stratification. NPC2, LY96, and TPP1, highly expressed in TAMs, were implicated in promoting tumor growth and immune escape, offering potential targets for novel therapeutic interventions.

Ganmai Dazao Decoction (GMZ) is a famous classic traditional Chinese medicine compound prescription for the treatment of depression and has a good clinical effect. However, its specific components and molecular mechanisms are still unclear. The chronic unpredictable mild stress (CUMS) mouse model was employed in this study, and the antidepressant effect of GMZ was evaluated via behavioral and serum corticosterone level assessment. Chemical components in GMZ water decoction, drug-containing serum, and drug-containing brain homogenate were detected by ultraperformance liquid chromatography quadrupole time-of-flight mass spectrometry (LC-TOF-MS). GO and KEGG analyses were performed on the intersecting targets of drugs and diseases to predict the antidepressant pharmacological mechanism of GMZ. Potential active components of GMZ were screened by molecular docking. The predicted targets were further validated for their role in and to verify the pharmacological mechanism of GMZ through <i>in vitro</i> and <i>in vivo</i> experiments. The CUMS mouse model was successfully established and GMZ (7 and 14 g/kg) was proven to have an antidepressant effect in this study. By using LC-TOF-MS analysis, 36 chemical components were identified in GMZ water decoction, 33 and 7 chemical components were identified in mouse serum and brain homogenates, respectively. There were 237 intersection targets between drug active targets and disease targets, and 48 core targets were further screened by the STRING database. GO and KEGG analyses indicated that GMZ regulated cAMP signaling pathway to achieve an antidepressant effect. Three potential components of GMZ were screened out by molecular docking, including Glyasperin C, Naringenin, and Scopoletin. The regulation effects of GMZ and its active components on the cAMP-PKA-CREB-BDNF signaling pathway were confirmed by <i>in vitro</i> and <i>in vivo</i> experiments. Our research proved that GMZ exerts an antidepressant effect through regulating the cAMP signaling pathway based on the model of "pharmacodynamics-serum pharmacology-network pharmacology-mechanism verification". This model could be a prospective approach to exploring the pharmacological mechanisms of traditional Chinese medicine.

<h4>Introduction</h4>Previous studies have reported that genetic polymorphisms may impact the signs and symptoms of temporomandibular disorder (TMD). Therefore, this study aimed to investigate the association between polymorphisms in the <i>Dopamine Receptor D2</i> (<i>DRD2</i>) and <i>Ankyrin Repeat and Kinase Domain Containing 1</i> (<i>ANKK1</i>) genes and oral health-related quality of life of male patients with TMD.<h4>Methods</h4>This cross-sectional study included construction workers with at least one sign or symptom of TMD. The reduced version of the Oral Health Impact Profile questionnaire (OHIP-14) was used to assess oral health-related quality of life. Genomic DNA was used to genotype genetic polymorphisms in the locus 11q22-q23, one in <i>ANKK1</i> (rs1800497) and two in <i>DRD2</i> (rs6275 and rs6276), using real-time polymerase chain reaction. The total OHIP-14 score and those for each domain were compared among the genotypes using the Kruskal-Wallis test and Dunn's test in the genotypic co-dominant model. The Mann-Whitney test was used in the recessive model (alpha = 0.05).<h4>Results</h4>The sample included a total of 114 male patients. OHIP-14 total score ranged from 0 to 33. Chronic pain (87.7%), followed by disc displacement (38.2%), was the most common sign and symptom observed. All the genetic polymorphisms assessed were within the Hardy-Weinberg equilibrium. The "Handicap" domain (D6) was statistically associated with the genetic polymorphism rs1800497 in <i>ANKK1</i> (<i>p</i> = 0.008). The genetic polymorphism rs1800497 Taq1A in <i>DRD2/ANKK1</i> was associated with oral health-related quality of life, as measured by the handicap domain in OHIP-14, in male patients with TMD.<h4>Discussion</h4>This study showed that genetic polymorphisms can negatively impact the oral health-related quality of life, as measured by the handicap domain of the OHIP-14. The physical and emotional condition of patients, together with biological pathways, should receive more attention in future studies, and personalized treatment plans should be created to improve patients' quality of life.

Peroxisomes are essential organelles, present in all nucleated cells, with key roles in lipid and redox homeostasis. They are important for maintaining healthy cell function, with defects in peroxisome biogenesis and/or metabolism leading to disease. Notably, patients with peroxisomal diseases exhibit predominantly neurological phenotypes, and peroxisomes are observed to be altered in a range of neurodegenerative conditions, highlighting the crucial roles they play in the brain. While most studies so far have focused on the contribution of peroxisomal metabolism, it is becoming apparent that many different aspects of peroxisome biology are necessary for healthy neural function. Peroxisomes are highly dynamic, responding to cellular needs with changes in number, shape and distribution. Furthermore, they do not act in isolation but instead interact and cooperate with a range of organelles to carry out their roles. This review summarizes our current knowledge on the importance of peroxisome dynamics and inter-organelle interactions in neuronal function and dysfunction. It considers their impact on neuronal physiology, and discusses the evidence that defects in these processes are associated with neurological pathophysiology and may thus represent a novel therapeutic target for treating diseases affecting the nervous system. Finally, the review outlines the current knowledge gaps relating to the mechanisms by which peroxisome dynamics and inter-organelle interactions influence neuronal (dys)function, proposing potential new research directions to address these and further our understanding of the multi-faceted roles peroxisomes play in brain health and disease.

<b>Background:</b> The STING (Stimulator of Interferon Genes) pathway plays a vital role in the body's innate immune defense system, primarily involved in DNA sensing and type I interferon production. While STING is well-established in various immune cells, its role in natural killer (NK) cells, particularly within the context of liver fibrosis, remains inadequately explored. <b>Aim:</b> The current study investigates the relationship between STING expression, NK cell activity, and insulin receptor (IR) signaling in patients with metabolic dysfunction-associated steatohepatitis (MASH). <b>Methods:</b> Peripheral NK cells were isolated from healthy controls and MASH patients with varying stages of liver fibrosis (early: F1/F2; advanced: F3/F4). The expressions of STING, IR, NK cell activation markers (CD107a, NKp46), and NK cell inhibitory markers (LAIR-1, Siglec-7) were assessed using flow cytometry. NK cell cytotoxicity against primary hepatic stellate cells (pHSCs) was evaluated through apoptosis assays. STING agonists (2'3'-cGAMP and DMXAA) were used to stimulate NK cells, and their effects on STING expression, NK cell activation, and cytotoxicity were measured. Additionally, the impact of insulin signaling on STING expression and NK cell function was examined. <b>Results:</b> Our results demonstrate that STING expression in NK cells correlates with disease severity in liver fibrosis. NK cells from MASH patients with advanced fibrosis (F3/F4) showed inhibited STING protein levels that were statistically comparable to healthy NK cells and accompanied by impaired cytotoxicity and decreased IFN-γ production. In contrast, NK cells from early fibrosis (F1/F2) exhibited higher STING expression and better functional activity. STING agonist treatment (2'3'-cGAMP) restored STING expression and enhanced NK cell activity across all fibrosis stages. Furthermore, insulin treatment and combined insulin and 2'3'-cGAMP treatment synergistically upregulated both IR and STING expressions, leading to improved NK cell function and increased cytotoxicity, particularly in advanced fibrosis. <b>Conclusion:</b> Our results highlight the potential of targeting STING and insulin signaling pathways as a therapeutic approach in restoring NK cell function and enhance immune surveillance in liver fibrosis.

Increasing evidence underlines the role of B-cells in the development of hepatic fibrogenesis following viral infections and metabolic dysfunction, through different mechanisms depending on the etiology. Circulating biomarkers of B-cell activation-such as B-cell activating factor (BAFF), immunoglobulin G (IgG) subclasses, and free light chains (FLCs)-may be associated with different results between viral and metabolic hepatic fibrosis, supporting their use as diagnostic tools. We conducted a case-control study including 100 patients with liver fibrosis, 50/100 of metabolic etiology and 50/100 of viral etiology. A reference group of 30 healthy donors was included as control. Serum levels of BAFF were measured using ELISA, while IgG subclasses (IgG1, IgG2, IgG3, IgG4), κ-FLC, λ-FLC, and the κ/λ ratio were quantified by turbidimetric methods. In univariate analysis, κ-FLC, λ-FLC, and BAFF levels were significantly elevated in both patient groups, with the highest concentrations consistently observed in metabolic fibrosis. IgG2 was selectively increased in metabolic fibrosis, whereas IgG3 was specifically elevated in viral fibrosis. Multivariate analysis confirmed these findings, showing a clear clustering of the three groups and identifying increased BAFF and κ-FLC as key features of metabolic fibrosis, while elevated IgG3 emerged as the most distinctive marker of viral etiology. These results reveal distinct B-cell-related immunological signatures in metabolic and viral hepatic fibrosis supporting the role of BAFF, FLCs, and IgG subclasses as biomarkers of etiological differentiation, and provide novel insights into the immune mechanisms driving fibrosis progression, potentially contributing to the identification of new therapeutic targets.

<h4>Background</h4>Hepatocellular carcinoma (HCC)-a predominant type of primary liver cancer-poses a significant global health threat with high incidence and mortality rates. Despite advances in treatment modalities, including surgery, chemotherapy, and immunotherapy, HCC exhibits high relapse rates and low long-term survival, necessitating the identification of novel prognostic markers and treatment targets. This study aims to develop a prognostic model centered on the PDZ domain by identifying key PDZ proteins associated with HCC through bioinformatics analysis of large-scale public datasets, in order to improve prognosis prediction and inform therapeutic strategies.<h4>Methods</h4>Differentially expressed PDZ proteins (DEPs) in HCC were identified through RNA sequencing (RNA-seq) analysis from The Cancer Genome Atlas (TCGA), International Cancer Genome Consortium (ICGC), and Gene Expression Omnibus (GEO) databases. Cox regression and random survival forest (RSF) modeling were employed to construct a prognostic model and evaluate the prognostic potential of DEPs. Subsequently, the correlation of DEP-related risk scores with immune cell infiltration and genetic variations was analyzed separately. Quantitative polymerase chain reaction (qPCR) was conducted to validate the expression of key DEPs in HCC tissues.<h4>Results</h4>A prognostic model for HCC constructed using nine key DEPs demonstrated reliable predictive performance across 1-, 3-, and 5-year survival rates. DEP-related risk scores were significantly associated with immune cell infiltration, with high-risk groups exhibiting an increase in pro-tumor immune cells and a decrease in anti-tumor immune cells. Genetic variations, including single nucleotide polymorphisms (SNPs) and copy number variations (CNVs), also differed between high- and low-risk groups. qPCR validation confirmed that the expression of SNX27, DLG5, PARD3, and RHPN1 was significantly upregulated in HCC tissues.<h4>Conclusions</h4>PDZ proteins may serve as prognostic markers and therapeutic targets in HCC. DEP-related risk scores offer insights into immune infiltration patterns and treatment responsiveness, providing a foundation for future HCC research and development of precision medicine.

Hydrocephalus is a complex neurological condition marked by abnormal cerebrospinal fluid (CSF) accumulation, often leading to elevated intracranial pressure and structural brain damage. Despite advances in surgical treatment, diagnostic precision and prognosis remain challenging, especially in idiopathic normal pressure hydrocephalus (iNPH). This narrative review aims to synthesize the current knowledge regarding molecular and neuroimaging biomarkers that hold diagnostic, prognostic, and therapeutic significance in hydrocephalus. A comprehensive literature search was conducted across PubMed, Scopus, Web of Science, and Google Scholar. The inclusion criteria encompassed peer-reviewed studies involving congenital or acquired hydrocephalus and reporting on mechanistic, diagnostic, or monitoring biomarkers. Both established and emerging biomarkers were included, and preclinical findings were considered when translational relevance was apparent. The review highlights a broad spectrum of molecular markers including aquaporins, vascular endothelial growth factor, neurofilaments, glial fibrillary acidic protein, matrix metalloproteinases, and neuroinflammatory markers. The genetic markers associated with ciliogenesis also show promise in subtyping disease. Parallel to molecular advances, neuroimaging techniques, ranging from classic markers like Evans' index to advanced modalities such as diffusion tensor imaging (DTI), arterial spin labeling (ASL), and glymphatic MRI, provide functional perspectives on hydrocephalus diagnosis and management, while artificial intelligence may further enhance diagnostic algorithms. Molecular and imaging markers could not only increase diagnostic confidence, but also provide information on disease causes and progression. As research progresses, merging various methodologies may result in more accurate diagnoses.

Genetic dystonias are a heterogeneous group of movement disorders characterized by involuntary, sustained muscle contractions that cause repetitive movements and abnormal postures. Often beginning in childhood, they can significantly affect quality of life. Although individually rare, genetic causes are collectively relevant in pediatric dystonias, with over 250 associated genes. Among these, <i>TOR1A</i>, <i>SGCE</i>, and <i>KMT2B</i> are the most frequently reported in pediatric forms. Diagnosis is challenging due to the wide clinical and genetic variability. Recent advances in genetic testing, including whole-exome and whole-genome sequencing, have improved the early identification of causative variants. Functional data on selected mutations are helping to refine genotype-phenotype correlations. Management typically requires a multidisciplinary approach. Symptomatic treatments include anticholinergics, benzodiazepines, and botulinum toxin, while deep brain stimulation can be effective in refractory cases, especially in patients with <i>TOR1A</i> variants. Disease-modifying therapies are also emerging, such as gene therapy for AADC deficiency, highlighting the potential of precision medicine. This review provides an updated overview of pediatric genetic dystonias, with a focus on differential diagnosis and treatment strategies. Early and accurate diagnosis, together with personalized care, is key to improving outcomes in affected children.

The sterol demethylation inhibitors (DMIs) are among the most widely used fungicides for controlling black Sigatoka (<i>Mycosphaerella fijiensis</i>) and yellow Sigatoka (<i>Mycosphaerella musicola</i>) in banana plantations in Brazil. Black Sigatoka is considered more important due to causing yield losses of up to 100% in commercial banana crops under predisposing conditions. In contrast, yellow Sigatoka is important due to its widespread occurrence in the country. This study aimed to determine the current sensitivity levels of <i>Mf</i> and <i>Mm</i> populations to DMI fungicides belonging to the chemical group of triazoles. Populations of both species were sampled from commercial banana plantations in Registro, Vale do Ribeira, São Paulo (SP), Ilha Solteira, Northwestern SP, and Janaúba, Northern Minas Gerais, and were further characterized phenotypically. Additionally, allelic variation in the <i>CYP51</i> gene was analyzed in populations of these pathogens to identify and characterize major mutations and/or mechanisms potentially associated with resistance. Sensitivity to the triazoles propiconazole and tebuconazole was determined by calculating the 50% inhibitory concentration of mycelial growth (EC₅₀) based on dose-response curves ranging from 0 to 5 µg mL^-1. Variation in sensitivity to fungicides was evident with all nine <i>Mf</i> isolates showing moderate resistance levels to both propiconazole or tebuconazole, while 11 out of 42 <i>Mm</i> strains tested showed low to moderate levels of resistance to these triazoles. Mutations leading to CYP51 substitutions Y136F, Y461N/H, and Y463D in <i>Mm</i> and Y461D, G462D, and Y463D in <i>Mf</i> were associated with low or moderate levels of resistance to the triazoles. Interestingly, Y461H have not been reported before in <i>Mm</i> or <i>Mf</i> populations, and this alteration was found in combination with V106D and A446S. More complex CYP51 variants and <i>CYP51</i> promoter inserts associated with upregulation of the target protein were not detected and can explain the absence of highly DMI-resistant strains in Brazil. Disease management programs that minimize reliance on fungicide sprays containing triazoles will be needed to slow down the further evolution and spread of novel CYP51 variants in <i>Mf</i> and <i>Mm</i> populations in Brazil.

No abstract available.

<h4>ABSTRACT</h4> Approved by the FDA in 2006, varenicline became the first nicotinic-based therapeutic for smoking cessation and has since been used by tens of millions of smokers worldwide. Varenicline works by targeting the α4β2 nicotinic acetylcholine receptor (nAChR), the primary focus for nicotine addiction, where ligand recognition by the receptor triggers ion channel opening. While widely recognized that varenicline’s development was rooted in the well-established pharmacology of cytisine, the two compounds display notably different profiles, not only at nAChRs, but also at key off-target sites such as the 5-HT 3 serotonin receptor. Despite varenicline’s widespread use and proven efficacy as a smoking cessation aid, our knowledge of the precise molecular mechanism underlying its action, particularly the specific receptor-ligand interactions that underpin its functional specificity, remains incomplete. Through a multidisciplinary approach that integrates complementary fields of research, this study reveals the critical receptor-ligand interactions that distinguish varenicline from related nAChR agonists, such as cytisine and nicotine. Our findings reveal previously unrecognized, critical hydrogen bonding interactions within the α4β2 binding sites, specifically involving α4T139, α4T183, and β2S133, that are uniquely and selectively engaged by varenicline. Of these, β2S133 emerged as the pivotal determinant of varenicline’s function, with substitution by valine significantly impairing the ligand efficacy. Furthermore, the design and synthesis of novel varenicline analogues shed new light into the functional importance of the ligand’s quinoxaline moiety, revealing that not just the presence but also the precise positioning of this hydrogen bond acceptor are critical for receptor activation by varenicline. Together, these findings uncover a previously uncharacterized interaction network essential for varenicline’s function at α4β2, offering a deeper and more comprehensive framework for understanding its distinct pharmacological profile while expanding our broader understanding of how ligand binding is translated into function in these receptors. <h4>GRAPHICAL ABSTRACT</h4>

Studying the genetics of intelligence can help us understand the neurobiology of cognitive function and the aetiology of rare neurodevelopmental conditions. The largest previous genetic studies of intelligence have used ~270k individuals who completed the fluid intelligence (FI) tests in UK Biobank. Here, we integrate additional FI measures in this cohort and leverage 82 correlated variables to impute FI values for unmeasured individuals, increasing the sample size to >450k. Through population-based and within-family genome-wide association studies and genetic correlation analyses, we show that this imputation produces a phenotype that genetically resembles measured FI, as well as reduces ascertainment bias within the cohort. We further show that combining measured and imputed FI scores increases the number of independent SNP associations (p<5x10-8) from 385 to 608 and increases polygenic score accuracy in external cohorts by 15% on average. Additionally, incorporating imputed FI scores increases the number of gene-level associations with rare variants from five to twenty-six (FDR<1%). These include fourteen well-established developmental disorder-associated genes, a four-fold enrichment (p=8x10-8); for several of these (e.g. ATP1A1, CACNA1E), our results suggest that the gene has a loss-of-function mechanism as well as the previously-documented altered function mechanism. We also implicate twelve genes without strong prior evidence of association developmental disorders, of which eight have not been previously linked to intelligence (ROBO2, RB1CC1, ANK3, CHD9, TLK1, PCLO, DPP8, IPO9). These twelve genes were significantly enriched for de novo loss-of-function mutations in a set of >31k patients with developmental disorders (p=6.8x10-4). We further identify three genes showing significant rare variant associations with educational attainment but not with FI in UK Biobank, including CADPS2 in which, unusually, protein-truncating variants show a positive association. Our results demonstrate the power of phenotype imputation for genetic studies and suggest that incorporating genetic association results for cognitive phenotypes in the general population could help discover new developmental disorder genes.

<h4>Background</h4> Sexual maturation is a key developmental process important for reproductive success. Understanding the molecular mechanisms behind variation in sexual maturation can provide insights into reproductive biology and how life history variation is encoded in the genome. Atlantic salmon ( Salmo salar ) has become an excellent sexual maturation research model due to its diversity of life history strategies and its ecological and economic importance. A major challenge has been the lack of a comprehensive transcriptional investigation of reproductive tissues that captures the dynamic transcriptional changes across individuals, tissues, and developmental stages. Long non-coding RNAs (lncRNAs) also play crucial roles in maturation, yet their functions in salmon maturation remain underexplored. <h4>Results</h4> In this study, we sequenced 98 transcriptomes and found substantial transcriptomic complexity in the gonad and pituitary tissues of Atlantic salmon. We identified transcripts corresponding to 2,364 putative newly characterized protein-coding genes and 4,421 putative long intergenic non-coding RNAs (lincRNAs), many with tissue-specific expression. Gene co-expression network analysis (WGCNA) revealed tissue-specific gene network modules, linked to GO terms including Wnt signaling in immature testis, lipid metabolism, and cilia assembly in mature testis, ribosome biogenesis and DNA repair in the ovary, and hormone activity in pituitary. We identified new copies of known genes, such as gh1, pou3f2 , and ier5 associated with the regulation of gonadal and pituitary functions. Some lncRNAs and their nearest genes showed correlated expression within modules, suggesting potential regulatory roles. Candidate lincRNAs indicated cis-acting regulatory potential on genes like tnfrsf11b and fgl1 , which are implicated in immune privilege during gonadal development and sperm quality control. <h4>Conclusions</h4> Our study provides a comprehensive transcriptomic analysis of Atlantic salmon gonad and pituitary tissues, significantly improving the functional annotation of the Atlantic salmon genome. These findings reveal key regulatory pathways and novel molecular players involved in sexual maturation, particularly in the testis. Importantly, our study highlights the regulatory potential of lncRNAs in reproductive biology and maturation age variation, advancing our understanding of the molecular mechanisms governing sexual maturation. They further unlock future gene expression analyses and regulatory network reconstruction for dissecting the roles of lncRNAs in Atlantic salmon life history variation.

CRISPR is a gene editing tool adapted from naturally occurring defense systems from bacteria. It is a technology that is revolutionizing the interrogation of gene functions in driving liver disease, especially through genetic screens and by facilitating animal knockout and knockin models. It is being used in models of liver disease to identify which genes are critical for liver pathology, especially in genetic liver disease, hepatitis, and in cancer initiation and progression. It holds tremendous promise in treating human diseases directly by editing DNA. It could disable gene function in the case of expression of a maladaptive protein, such as blocking transthyretin as a therapy for amyloidosis, or to correct gene defects, such as restoring the normal functions of liver enzymes fumarylacetoacetate hydrolase or alpha-1 antitrypsin. It is also being studied for treatment of hepatitis B infection. CRISPR is an exciting, evolving technology that is facilitating gene characterization and discovery in liver disease and holds the potential to treat liver diseases safely and permanently.

The ability to assay the molecular composition of biological systems with single-cell resolution has revolutionized our understanding of tissue heterogeneity and function. Recent advances in single-cell proteomics (SCP) now enable the unbiased quantification of the proteome to a depth of several thousand proteins across hundreds of cells. Yet, broader adoption beyond specialized groups remains limited due to the need for specific equipment and expertise. A major challenge in making these analyses more broadly available is sample preservation for the transport of biological material to SCP-capable facilities. To address this issue and provide practical solutions, we first evaluated various cell preservation methods from monolayer culture samples, then tested our optimized methodology on both cultured cells and, for the first time, preserved animal tissue from an <i>in vivo</i> mouse model. Our findings highlight that the feasibility of SCP analyses in preserved tissues is more likely to be successful, significantly expanding its current applicability. By optimizing upstream processing, our approach enables robust single-cell proteome analysis of both cells and tissues, making SCP more accessible to the wider scientific community. Ultimately, this advancement expands the potential applications of SCP, particularly in disciplines where analyzing rare or heterogeneous populations is beneficial.

RNA therapeutics represent a disruptive technology that has transformed drug discovery and manufacturing, gaining significant prominence during the COVID-19 pandemic. RNA therapeutics encompass diverse molecules like antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), microRNAs (miRNAs), RNA aptamers, and messenger RNAs (mRNAs), which can function through different mechanisms. RNA therapeutics are increasingly used to treat various diseases, including neurological disorders. For example, ASO therapies such as nusinersen for spinal muscular atrophy and eteplirsen for Duchenne muscular dystrophy are successful applications of RNA-based treatment. Emerging ASO treatments for Huntington's disease and amyotrophic lateral sclerosis are also promising, with ongoing clinical trials demonstrating significant reductions in disease-associated proteins. Still, delivery of these molecules remains a pivotal challenge in RNA therapeutics, especially for ASOs in penetrating the blood-brain barrier to target neurological disorders effectively. Nanoparticle-based formulations have emerged as leading strategies to enhance RNA stability, reduce immunogenicity, and improve cellular uptake. Despite these advances, significant hurdles remain, including optimizing pharmacokinetics, minimizing off-target effects, and ensuring sustained therapeutic efficacy. Regulatory frameworks are evolving to accommodate the unique challenges of RNA-based therapies, including ASOs with efforts underway to establish comprehensive guidelines for RNA therapeutics, yet there are also sustainable manufacturing issues that need to be considered for long-term feasibility. By addressing these challenges, RNA therapeutics hold immense potential to revolutionize treatment paradigms for neurological disorders. Looking forward, the future of RNA therapeutics in neurology appears promising but requires continued interdisciplinary collaboration and technological innovation.

Anisotropic hydrogels are promising candidates as load-bearing materials for tissue engineering, while huge challenges remain in exploring effective and scalable methods for the preparation of anisotropic hydrogels with simultaneous high tensile strength, large toughness, good fracture strain, excellent fatigue and swelling resistances. Inspired by the brick-and-mortar layered structure of nacre and the hierarchical fibril strucure of soft tissues (e.g., tendon and ligament), a facile organogel-assissted calendering strategy is reported to design anisotropic hydrogels with a highly oriented and dense fiber lamellar strucure. The synergy of shearing and annealing promotes macromolecular chain alignment and crystallinity along the calendering direction while forming a nacre-like lamellar morphology in the thickness direction. The tensile strength, elastic modulus, toughness and fracture energy of the anisotropic hydrogels can reach as high as 41.0 ± 6.4 MPa, 67.0 ± 5.1 MPa, 46.2 ± 3.3 MJ m^-3, and 62.20 ± 8.55 kJ m^-2, respectively. More importantly, the hydrogels show excellent crack growth and swelling resistances with the fatigue threshold increased to 2170 J m^-2. This study provides a promising approach for fabrication of large-sized biomimetic anisotropic hydrogels with outstanding mechanical properties for biomedical and engineering applications.

<h4>Background</h4>CAR-T cell therapy has demonstrated remarkable success in hematologic malignancies; however, its effectiveness against solid tumors remains limited due to tumor antigen heterogeneity. NKG2DLs, including MICA/B and the ULBP family, are stress-induced molecules frequently upregulated on the surface of tumor cells and components of the tumor microenvironment, providing attractive targets for immunotherapy. To broaden the targeting capability beyond conventional Claudin18.2-directed CAR-T cells, we engineered a Synthetic NKG2D Receptor (SNR). The SNR comprises the extracellular domain of NKG2D fused with the intracellular signaling domains of DAP10 and DAP12, enabling effective targeting of NKG2D ligands (NKG2DLs).<h4>Methods</h4>Expression of NKG2DLs and CLDN18.2 were detected by immunohistochemistry on a gastric cancer tissue microarray. We designed SNR CAR-T cells by linking CLDN18.2 CAR with SNR by a 2A self-cleaving peptide. We assessed their cytotoxicity, tumor infiltration, persistence, and antitumor efficacy using in vitro assays, patient-derived xenograft (PDX) models, and murine syngeneic models. Additionally, transcriptomic analysis and flow cytometry were performed to evaluate exhaustion and memory markers.<h4>Results</h4>SNR CAR-T cells demonstrated enhanced cytotoxicity against tumor cells with heterogeneous CLDN18.2 expression, effectively lysing both CLDN18.2-positive and NKG2DL-positive tumor cells in vitro. In PDX and murine models, SNR CAR-T cells exhibited superior antitumor efficacy, leading to significant tumor regression and CAR-T expansion compared to conventional CAR-T cells. Furthermore, SNR CAR-T cells displayed reduced expression of exhaustion markers and increased expression of memory-associated markers. Enhanced tumor infiltration, proliferation and cytotoxicity within the tumor microenvironment, and a reduced presence of myeloid-derived suppressor cells (MDSCs) and tumor neovasculature were observed. Importantly, SNR CAR-T cell therapy was well-tolerated, with no significant toxicity noted in all the treated animals.<h4>Conclusion</h4>The SNR CAR-T cell approach addresses tumor antigen heterogeneity and suppressive tumor microenvironment, offering a promising therapeutic strategy for solid tumors and paving the way for its future clinical applications.

Hemophagocytic lymphohistiocytosis (HLH) has been described as a threshold disease depending on triggering factors and the residual cytotoxic capacity of NK cells. This study aimed to investigate the clinical characteristics of Epstein-Barr virus (EBV)-triggered primary HLH and the prognostic value of EBV-DNA load in EBV-triggered primary HLH cases. We retrospectively analyzed the clinical data of 95 patients with primary HLH treated between January 2013 and January 2024. Based on the peripheral blood EBV status at initial diagnosis, 57 patients were categorized into the EBV-triggered primary HLH group and 38 patients into the non-EBV-triggered primary HLH group. Clinical and functional characteristics, response to treatment, and prognosis were compared between the two groups. Among patients with EBV-triggered primary HLH, the proportion of patients with familial HLH type 2 (FHL2) was significantly lower (P = 0.011), whereas the proportion of patients with X-linked lymphoproliferative disorder (XLP) was higher (P = 0.037). Functional assays showed that in primary HLH patients with gene defects in cytotoxic degranulation, the proportion of EBV-triggered primary HLH patients with reduced NK cell activity and degranulation function was significantly higher (P = 0.026 and P = 0.030). Importantly, multivariate Cox regression analysis identified EBV-DNA > 10,000 copies/mL as an independent risk factor affecting the prognosis of patients with EBV-triggered primary HLH, particularly in non-FHL cases (P = 0.045). EBV-triggered primary HLH is more prevalent in patients with XLP but less frequent in FHL2 patients. High EBV-DNA load is an adverse prognostic factor in EBV-triggered primary HLH patients.