ABSTRACT Gastric cancer often presents at advanced stages, limiting treatment outcomes. Circulating tumor cells (CTCs), particularly those undergoing epithelial–mesenchymal transition (EMT), have emerged as promising biomarkers for real‐time disease monitoring. Using magnetic‐activated cell sorting (MACS), we enriched both epithelial (EpCAM+) and mesenchymal (CD90+) CTCs and evaluated the expression of the tumor‐suppressive microRNA miR‐29b in CTCs of gastric cancer patients. This study aimed to explore the clinical relevance of perioperative CTC phenotypes and miR‐29b as potential diagnostic and prognostic markers. We conducted a multi‐phase study integrating systematic review, bioinformatics, and experimental analyses to investigate CTC phenotypes and ECM‐associated gene expression in GC. Peripheral blood samples were collected from 30 treatment‐naïve gastric cancer patients at two time points: before initiation of therapy and 1 month after curative‐intent gastrectomy. Circulating tumor cells (CTCs) were isolated using CD45‐negative magnetic‐activated cell sorting (MACS). Immunofluorescence staining was performed to phenotype CTCs based on EpCAM (epithelial) and CD90 (mesenchymal) expression. Total RNA was extracted from tumor tissues and isolated CTCs to evaluate ECM‐related genes and miR‐29b expression using quantitative real‐time PCR. Bioinformatics‐guided gene selection and statistical analysis were used to assess diagnostic and prognostic relevance. CTCs were detected in 83.3% (25/30) of gastric cancer patients prior to treatment. Mesenchymal CTCs (CD90+) were markedly more frequent than epithelial CTCs (EpCAM+), with mean baseline counts of 14.07 versus 0.5 cells per 10 mL of blood, respectively. Following gastrectomy and chemotherapy, mesenchymal CTCs significantly declined (mean: 1.18; p = 0.0064), while epithelial CTCs were nearly undetectable (p = 0.0246). Based on perioperative CTC dynamics, patients were stratified into three risk groups: high‐risk (16%), intermediate‐risk (68%), and low‐risk (16%). All observed deaths occurred in the high‐risk group during 14 months of follow‐up, with Kaplan–Meier analysis showing significantly lower survival in this group (p = 0.0107). Expression analysis revealed significant downregulation of miR‐29b in tumor tissues (fold change = 0.5163; p = 0.0136) and in CTCs (fold change = 0.2049; p < 0.0001) compared to healthy controls. Although miR‐29b levels were broadly downregulated in gastric cancer patients, no significant differences were observed among clinicopathological subgroups within the patient cohort. ROC analysis showed strong diagnostic performance of miR‐29b in tissue (AUC = 0.883, 95% CI: 0.798–0.969), with 96.7% sensitivity and 70% specificity. CD90 serves as a sensitive mesenchymal marker for detecting CTCs that lack epithelial features, enabling more comprehensive identification of aggressive CTC phenotypes in gastric cancer. Moreover, the perioperative dynamics of CTCs—especially postoperative increases—provide a valuable tool for patient risk stratification and survival prediction. Importantly, our findings suggest that miR‐29b expression in CTCs, together with perioperative CTC dynamics, may serve as a non‐invasive biomarker for monitoring disease progression and recurrence in gastric cancer.
ABSTRACT Bicaudal C Homolog 1 (BICC1) is a conserved RNA‐binding protein that, in mammals, has been primarily associated with polycystic kidney disease and renal organogenesis. However, its role in other disease contexts, including cancer, remains poorly understood. In this study, we characterized the BICC1 interactome, with emphasis on its dependence on RNA and the sterile alpha motif (SAM) domain, to identify novel biological processes associated with BICC1 function. Protein complexes were purified from HEK293T cells by co‐immunoprecipitation and analyzed by mass spectrometry. Notably, co‐immunoprecipitations performed in the presence of RNA yielded a larger number of interacting proteins, with 31 of 71 proteins (~43%) uniquely identified under RNA‐preserved conditions, highlighting the critical role of RNA in mediating BICC1 protein–protein interactions. Enriched proteins were predominantly associated with mRNA splicing, the PRMT5 methylosome complex, and membraneless organelles, such as biomolecular condensates. Consistent with these findings, immunofluorescence assays performed on stressed cells revealed the co‐localization of BICC1 with stress granule markers. Moreover, BICC1 interactions with PRMT5, STK38, PARP1, and IGF2BP1 were confirmed by immunoblotting.
Emerging evidence indicated that long non-coding RNAs (lncRNAs) play critical roles in mammalian reproductive physiology, yet their roles in asthenozoospermia (AS), a major contributor to male infertility, remain elusive. Here, we identify LINC00966 as a candidate lncRNA associated with impaired sperm function. LINC00966 expression was significantly elevated in semen from AS patients and negatively correlated with sperm concentration, total motility, and progressive motility. In mouse testes, Linc00966 was detected in spermatogonia, spermatocytes and spermatids with predominant cytoplasmic localization as well as in GC-2spd cells. Furthermore, Linc00966 overexpression reduced cell viability and migration and increased intracellular reactive oxygen species (ROS), whereas knockdown enhanced viability and migration and lowered ROS in GC-2spd cells. Bulk RNA-seq analysis revealed that the phosphodiesterase 2A (Pde2a), a key cGMP/cAMP-hydrolyzing enzyme, were upregulated when Linc00966 was knockdown. The GO enrichment and KEGG analysis of differentially expressed genes suggested the cyclic guanosine monophosphate (cGMP) process were enriched. Further validation found that Linc00966 suppressed Pde2a. In clinical semen samples, LINC00966 and PDE2A expression levels were inversely correlated, and both were closely associated with semen quality parameters. Collectively, these findings implicated LINC00966 and its downstream target PDE2A as potential pathogenic regulators of sperm motility in AS. By linking lncRNA-mediated modulation of cGMP signaling and oxidative stress to spermatogenic cell function, our results demonstrated a plausible signaling axis underlying AS and suggests LINC00966/PDE2A as promising candidates for biomarker development and therapeutic investigation in male infertility.
ABSTRACT The COVID19 pandemic, caused by the betacoronavirus SARS‐CoV‐2, now ranks among the world's deadliest plagues, and certainly one of the most significant public health crises in the last 100 years. Despite more than 6 years of investigating the pandemic, it has been difficult to assess the origin of the disease in the City of Wuhan, China. The Wuhan Municipal Public Security Bureau regretfully suppressed information exchanges between physicians early in the Wuhan outbreak, which interfered with the contact tracing necessary for epidemiological investigations. The scientific community and government agencies throughout the world have been divided in their assessments of the pandemic's origin in Wuhan China. In the 2021 intelligence assessment released by the U.S. Office of the Director of National Intelligence (ODNI), only the FBI assessed the possibility of a laboratory origin. By 2023 the U.S. Department of Energy (DOE) revised its assessment to acknowledge a lab origin was likely, and in 2025 the U.S. Central Intelligence Agency (CIA) acknowledged a similar view. There has been political polarization of views, condescension and accusations of conspiracies among scientists. The COVID19 pandemic is prompting policy changes to research guidelines and recommendations for research restrictions based upon the assessment that pandemic may have resulted from laboratory research. While some express this as politically motivated, as we approach the seventh year of the COVID19 pandemic, more evidence has emerged that supports the research origin theory and scant evidence supporting a zoonotic emergence. It is imperative that our policy makers understand the facts when considering legislation and policies that could affect infectious disease research.
ABSTRACT The epidermis provides the body's outermost barrier, yet how G‐protein‐coupled receptor (GPCR) signaling via the G12/13 family regulates epidermal homeostasis in vivo remains unclear. Here, we selectively activated G12 signaling in keratinocytes using a chemogenetic strategy. Activation of a G12‐coupled designer receptor (G12D) in mouse epithelial cells induced pronounced epidermal thickening while preserving stratified architecture and avoiding overt inflammatory skin changes. This thickening was accompanied by increased Ki67‐positive cells, expansion of keratin 10‐ and filaggrin‐positive layers, and transcriptomic upregulation of genes related to keratinocyte differentiation, keratinization, and epidermal barrier function. Functionally, G12D activation strengthened barrier performance, as shown by blunted transepidermal water loss responses to mechanical barrier disruption. Although alarmin‐related genes were upregulated, cytokine analyses indicated only modest inflammatory changes. Pharmacologic inhibition of TYK2 (deucravacitinib) partially reduced G12D‐driven epidermal thickening, whereas mTORC1 inhibition (rapamycin) produced a stronger suppressive effect, suggesting that the mTORC1‐dependent keratinocyte response is a major driver of this phenotype, with a moderate TYK2‐dependent component. Together, these findings identify epidermal G12 signaling as a regulator that promotes “non‐pathological” epidermal thickening coupled to enhanced barrier function, supporting G12‐coupled GPCRs as potential therapeutic entry points for barrier‐compromised skin disorders.
ABSTRACT N 6 ‐methyladenosine (m 6 A), N 1 ‐methyladenosine (m 1 A), 5‐methylcytosine (m 5 C), 7‐methylguanosine (m 7 G), and pseudouridine (ψ) have been identified as the most prevalent epitranscriptomic modifications on eukaryotic RNAs. It is not yet clear about the comprehensive analysis of RNA m 6 A/m 1 A/m 5 C/m 7 G/ψ in pancreatic cancer, one of the most aggressive malignancies all over the whole world. We have identified that most of the RNA m 6 A/m 1 A/m 5 C/m 7 G/ψ regulators are highly expressed in pancreatic cancer. Copy number variations (CNVs) or the methylation level of some genes are associated with their mRNA expression and affect the overall survival of patients. The expression of most of the regulators is positively related to the immune cell infiltration of the tumor microenvironment (TME). Furthermore, the regulators or TME‐infiltrated cells are associated with tumor heterogeneity or tumor stemness. According to the similarity of TME‐infiltrating cells, we classify patients into two different clusters, and we find that patients in cluster 1 (C1) are at high risk, have worse prognoses, and are more sensitive to drug treatment than cluster 2 (C2). Three machine learning algorithms, least absolute shrinkage and selection operator (LASSO), support vector machine‐recursive feature elimination (SVM‐RFE), and random forest (RF), show that some regulators have a strong correlation with survival. Our study has revealed the potential roles of regulators in pancreatic cancer, which may provide new insights into the field of pancreatic cancer research.
ABSTRACT Simulated microgravity (SMG or μG) influences Toxoplasma gondii growth and modifies the cellular structure and function of the host cells. As an obligate intracellular parasite, Toxoplasma gondii depends on the metabolic activity and mitochondrial function of its host. However, the impact of SMG on host–pathogen interactions remains unclear. We investigated the effect of SMG on host cell structure and mitochondrial functions and its impact on Toxoplasma gondii infection. Cultured human foreskin fibroblasts (HFF) and Vero cells under SMG formed stratified cell layers and exhibited morphological mitochondrial abnormalities, such as outer membrane distortion and inner membrane distension. Functional analyses revealed altered mitochondrial membrane potential and intracellular ATP levels without substantial changes in reactive oxygen species. Upon infection, T. gondii showed cell‐type‐dependent growth, increasing in Vero cells but decreasing in HFF at 48 h post‐infection. These findings indicate that SMG‐induced mitochondrial remodeling is associated with altered host cell susceptibility to toxoplasmosis. Our study highlights the role of physical environmental changes in modulating host–pathogen interactions in the context of host cell mitochondrial function.
ABSTRACT The influential 2019 study by Bycroft et al. reported remarkably pronounced ultrafine‐scale genetic structure within a small region of Galicia (northwestern Iberia). Using fineSTRUCTURE clustering of ChromoPainter coancestry profiles, the authors identified multiple internally coherent clusters characterized by high levels of within‐group haplotype sharing and reduced sharing with neighboring populations. On a broader national scale, they also reported a marked East–West genetic differentiation and North–South homogeneity across the Iberian Peninsula, patterns which they largely attributed to historical population movements associated with the gradual territorial expansion of Christian kingdoms between the 8th and 15th centuries following the initial Islamic conquest. Several subsequent studies have adopted similar interpretations, but all relied on the same genetic resource, the EPICOLON Phase I cohort. Following a detailed reexamination of Bycroft et al., we identify multiple methodological issues, most notably signals consistent with unaccounted batch effects in EPICOLON, some of which were recognized by the authors in earlier studies. In contrast, we highlight recent analyses based on independent genomic datasets that consistently report high levels of genetic homogeneity within Galicia and across central and southern Iberia, with no evidence supporting a pronounced East–West genetic divide across Iberia. In addition, recent studies have documented North African demographic influences in Iberia, particularly in Galicia, predating the period of Islamic rule and therefore challenging the historical interpretation proposed by Bycroft et al. We also show that while fineSTRUCTURE is a powerful tool for detecting subtle population stratification, it may be highly sensitive to technical confounders, which can magnify or distort inferred clustering patterns. This study demonstrates that technical artifacts can strongly affect fine‐scale population structure inferred by fineSTRUCTURE , emphasizing the need for rigorous quality control and cautious interpretation of highly resolved genetic clusters.
ABSTRACT Diabetes is a global health concern, with type 2 diabetes mellitus (T2DM) being the most common form. T2DM is characterized by insulin resistance and impaired glucose uptake in skeletal muscle. Spexin shows hypoglycaemic effects, but its mechanism of action, especially in skeletal muscles, remains elusive. This study aims to investigate the effects of spexin on key genes associated with the insulin‐dependent pathway in C2C12 cells. The differentiation of C2C12 myoblasts to C2C12 myotubes was confirmed using the fusion index and gene expression of myogenic factor 5 (Myf5), myosin heavy chain (MyHC), and myogenic regulatory factor 4 (MRF4) analysis. Cytotoxicity of spexin at varying concentrations (100 nM, 200 nM, 400 nM, 800 nM, and 1000 nM) was assessed via MTS assay. Insulin resistance–like conditions were induced in C2C12 myotubes using a high‐glucose medium and characterized at the mRNA level by quantitative reverse transcription PCR (RT‐qPCR) of peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha (PGC‐1α), glucose transporter 4 (GLUT4), and hexokinase II (HKII) genes. Fusion index analysis evaluated high‐glucose effects on myogenesis. RT‐qPCR examined spexin's effects on insulin receptor substrate 1 (IRS‐1), phosphatidylinositol 3‐kinase (PI3K), and GLUT4 expression over different time points. Spexin concentrations at 1000 nM were not cytotoxic. High‐glucose incubation did not affect myogenesis yet was associated with downregulation of PGC‐1α and GLUT4 genes, consistent with a transcriptional profile typically observed in insulin‐resistant states. Spexin‐treated C2C12 cells statistically significantly downregulated the expression of IRS‐1 after 2 h of incubation. The effects of spexin on IRS‐1, PI3K, and GLUT4 gene expression were comparable to metformin‐treated in insulin resistance C2C12 myotubes.
Heart failure (HF) as the final stage of cardiovascular disease in the elderly leads to frequent readmissions and seriously affects their quality of life. This study aimed to develop a predictive model for hospital readmission or death during the vulnerable phase in elderly HF patients, and to identify the key associated risk factors. The dataset was randomly divided into 70% training sets and 30% validation sets. Three feature selection methods were applied to the training data, followed by the construction of 18 predictive models using six machine learning (ML) algorithms (XGBoost, LightGBM, AdaBoost, GBDT, GNB, and SVM). The performance of each model was assessed on the validation set using receiver operating characteristic (ROC) curves, sensitivity, accuracy, specificity, F1 score, and Brier score. SHapley Additive exPlanations (SHAP) were used to interpret the feature contributions both globally and locally. Eleven models achieved an area under the ROC curve (AUC) greater than 0.8, with the Boruta-XGBoost model performing best, showing an AUC of 0.873 in the validation set, along with a sensitivity of 0.839, accuracy of 0.769, specificity of 0.747, F1 score of 0.634, and Brier score of 0.130. SHAP analysis revealed that the top five important features were hemoglobin (HGB), serum free thyroxine (FT4), age, diabetes, and serum potassium (K). The Boruta-XGBoost based risk prediction model, combined with SHAP interpretation, demonstrated high predictive accuracy and robust interpretability for forecasting hospital readmission or death during the vulnerable phase in elderly HF patients.
ABSTRACT Cigarette and hookah smoking remain prevalent worldwide, yet their comparative effects on oral microbiota and hematological parameters are not fully understood. This cross‐sectional study investigated these impacts among 87 healthy males (18–40 years) in Sulaymaniyah, Iraq, divided into cigarette smokers, hookah smokers, and non‐smokers (n = 29 each). Oral rinses were analyzed for microbial load and species distribution using selective media and standard identification techniques, while venous blood samples were evaluated for complete blood counts using an automated analyzer. Cigarette smokers exhibited the highest oral bacterial load (5.96 ± 0.19 log10 CFU/mL) compared with non‐smokers (3.70 ± 0.10; p < 0.001), followed by hookah smokers. Colonization by Candida albicans and Gram‐positive cocci (Staphylococcaceae, Streptococcaceae) was more frequent in smokers. Both smoking methods significantly increased RBC count, hemoglobin, and hematocrit levels (p < 0.01). Mean corpuscular volume increased significantly only in cigarette smokers, whereas red cell distribution width and mean platelet volume were significantly higher in hookah smokers (p < 0.01). Exploratory correlation analysis revealed strong positive associations between oral bacterial load and erythrocyte‐related parameters (RBC, HGB, HCT; r = 0.91–0.94), while weak correlations were observed with inflammatory and platelet indices (WBC, RDW, MPV). These findings indicate that cigarette and hookah smoking disrupt oral microbial balance and alter hematological parameters, with distinct patterns between smoking types. The observed correlations likely reflect parallel systemic effects of smoking‐related toxicants rather than a direct causal link between oral dysbiosis and erythropoiesis. Overall, the results reinforce that hookah smoking is not a safer alternative to cigarette use and highlight the need for targeted public health interventions.
ABSTRACT This study investigated the effects of lanthanum chloride on vascular calcification associated with chronic kidney disease (CKD) and the mechanisms involved in changes in nano‐hydroxyapatite. Vascular calcification was induced in CKD rats using a high‐phosphorus diet and adenine. Human vascular smooth muscle cells (hVSMCs) were calcified in vitro using sodium β‐glycerophosphate (β‐GP) and saturated nano‐hydroxyapatite. The effects of lanthanum chloride were evaluated using various analytical methods, including serum biochemistry, EVG, Vonkossa staining, Alizarin Red staining, Ca2+ detection, Western blotting, and transmission electron microscopy. The results showed that lanthanum chloride effectively inhibited calcium deposition and osteogenic differentiation, a finding confirmed in both in vivo and in vitro experiments. Scanning electron microscopy (SEM) analysis of the calcified crystals revealed altered crystal morphology and a decreased calcium‐to‐phosphorus ratio after treatment. X‐ray diffraction confirmed that these crystals were hydroxyapatite. Proteomic analysis indicated that the effects of lanthanum chloride were associated with apoptosis and the PPARγ/Wnt/β‐catenin signaling pathway. Lanthanum chloride inhibits apoptosis in human vascular smooth muscle cells (hVSMCs), while PPARγ inhibitors can reverse this effect. In summary, lanthanum chloride may significantly reduce vascular calcification in patients with CKD by altering the morphology of hydroxyapatite and activating PPARγ to inhibit apoptosis.
Acute inflammation is rapidly elicited in response to pathogens, toxins, and other cellular components that interact with immune cells through surface receptors, activating signaling cascades that lead to the expression of molecules aimed at neutralizing the pathogen and stimulating other cells to begin the healing process. However, a robust inflammatory response can be detrimental if it is prolonged. Therefore, the inflammatory response must be tightly regulated to prevent secondary harmful effects. In this regard, cells contain molecular switches that turn the inflammatory process on and off. The rapid response to infection and injury is mediated by constitutive cellular proteins that are activated by posttranslational modifications, of which phosphorylation is the most common. Consequently, compensatory mechanisms often involve the removal of phosphate groups by phosphatases. A family of phosphatases known as dual-specificity phosphatases (DUSPs), which dephosphorylate both tyrosine and serine/threonine residues, has emerged as a critical mechanism for controlling inflammation. To date, more than 40 DUSPs have been identified in the human genome. Despite their shared catalytic specificity, they appear to participate in diverse cellular processes and are expressed in response to various stimuli. In the present study, we detected a distinctive pattern of DUSP expression in response to exogenous lipid nanoparticles and toxins. These observations suggest that the transcriptional regulation of several DUSPs may be coordinated. They also raise the possibility that DUSPs may play redundant roles to ensure the proper resolution of the inflammatory response.
ABSTRACT Axon growth is an essential cellular process during neural development, and its dysregulation contributes to numerous neurodevelopmental disorders. During axon growth, extracellular signals direct neurons to extend projections that connect with their synaptic targets. Paxillin is a key member of adhesion sites that control motility by linking the intracellular actin cytoskeleton to the extracellular matrix. Paxillin also binds to the cytoskeletal protein, tubulin. However, little is known about the role of adhesion proteins in neurons. Here, we use conditional paxillin knockout mice to investigate how the loss of paxillin in pyramidal cortical neurons affects developing neuron morphology. Surprisingly, loss of paxillin in pyramidal cortical neurons caused no change in axon length or soma area between control (PxnF/F) and conditional paxillin knockout (PxnF/F; Emx1‐Cre) mice at basal conditions. Following brain‐derived neurotrophic factor stimulation, the loss of paxillin resulted in no change in soma area or axonal β‐tubulin levels, but did result in a significant increase in axon length, as compared to control. Finally, the corpus callosum size was not significantly different between PxnF/F and PxnF/F; Emx1‐Cre animals. In summary, these data suggest that paxillin is not required for axonal growth during neural development.
N6-methyladenosine (m6A), N1-methyladenosine (m1A), 5-methylcytosine (m5C), 7-methylguanosine (m7G), and pseudouridine (ψ) have been identified as the most prevalent epitranscriptomic modifications on eukaryotic RNAs. It is not yet clear about the comprehensive analysis of RNA m6A/m1A/m5C/m7G/ψ in pancreatic cancer, one of the most aggressive malignancies all over the whole world. We have identified that most of the RNA m6A/m1A/m5C/m7G/ψ regulators are highly expressed in pancreatic cancer. Copy number variations (CNVs) or the methylation level of some genes are associated with their mRNA expression and affect the overall survival of patients. The expression of most of the regulators is positively related to the immune cell infiltration of the tumor microenvironment (TME). Furthermore, the regulators or TME-infiltrated cells are associated with tumor heterogeneity or tumor stemness. According to the similarity of TME-infiltrating cells, we classify patients into two different clusters, and we find that patients in cluster 1 (C1) are at high risk, have worse prognoses, and are more sensitive to drug treatment than cluster 2 (C2). Three machine learning algorithms, least absolute shrinkage and selection operator (LASSO), support vector machine-recursive feature elimination (SVM-RFE), and random forest (RF), show that some regulators have a strong correlation with survival. Our study has revealed the potential roles of regulators in pancreatic cancer, which may provide new insights into the field of pancreatic cancer research.
ABSTRACT Retrotransposons, including non‐LTR elements such as LINEs and viral‐derived endogenous retroviruses (ERVs), have long been dismissed as “junk DNA” and thought to be biochemically inert. However, emerging evidence suggests that the erosion of epigenetic control during aging and pathological states can lead to the awakening of these dormant genetic elements. Although the role of ERVs in cancer and neurodegeneration is increasingly recognized, their impact on musculoskeletal health has received little attention. This perspective review synthesizes recent and previous findings linking retrotransposon reactivation (particularly ERVs) to osteoarthritis (OA), rheumatoid arthritis (RA), and potentially osteoporosis. We discuss the epigenetic mechanisms that typically silence ERVs in musculoskeletal tissues, how these mechanisms fail in disease, and how the resulting reactivation leads to viral and molecular mimicry. These processes trigger both innate and adaptive immune responses, as well as cellular senescence. Finally, we highlight the therapeutic potential of targeting retrotransposon dysregulation, including its encoded proteins and nucleic acid‐sensing pathways to treat chronic bone and joint disorders.
ABSTRACT Dapagliflozin is a sodium‐glucose cotransporter‐2 (SGLT‐2) inhibitor primarily used to treat type 2 diabetes by lowering blood glucose levels. In addition to its antidiabetic action, it has demonstrated cardioprotective and renoprotective effects, along with antioxidant, anti‐inflammatory, and anticancer activities. Encapsulation of dapagliflozin in nanocarriers represents an innovative strategy to improve existing therapies and develop targeted treatments. Such formulations can enhance solubility and stability, improve epithelial permeability and bioavailability, and reduce potential side effects. This study investigates the cellular and molecular effects of dapagliflozin encapsulated in chitosan nanoparticles, focusing on colon (Caco‐2) cells. The findings showed that free dapagliflozin significantly reduced cell viability, whereas the encapsulated form preserved high cell viability. In addition, the encapsulated drug effectively reduced reactive oxygen species levels, maintaining its antioxidant activity. Free dapagliflozin did not increase the expression of Nrf2, NFκB, or HO‐1 signaling pathways. In contrast, encapsulation in chitosan nanoparticles resulted in increased expression of all three pathways, indicating potential regulatory involvement in these signaling mechanisms.
ABSTRACT Apolipoprotein E4 (APOE4) is the strongest genetic risk factor for Alzheimer's disease (AD), yet it's unclear how this allele promotes disease. While factors like diet and sex may modify AD susceptibility in APOE4 carriers, the interaction between these factors is poorly understood. Here, we sought to determine if APOE4, sex, and diet interact to influence AD related outcomes in mice. Male and female APOE3 and APOE4 targeted replacement (TR) mice were fed a low‐fat diet or high‐fat diet from 4 to 8 months old. Serum neurodegenerative disease biomarkers, brain amyloid beta (Aβ), APOE, and tau, learning and memory, hippocampal mitochondrial function and proteomics data were collected. Serum GFAP and NfL were unaffected by APOE4, while HFD was associated with greater serum NfL and GFAP. Whole brain Aβ was significantly altered by sex, diet, and genotype. There was a main effect of genotype on levels of brain APOE with levels being lower in APOE4 mice. APOE4 TR mice also exhibited impaired learning before diet. Proteomic analysis revealed that APOE4 exerts diet‐ and sex‐dependent effects on mitochondrial pathways. This included downregulation of pyruvate metabolism in HFD males and oxidative phosphorylation in HFD females. Basal respiration was lower in APOE4 versus APOE3 TR females. We provide novel evidence that APOE4 may drive early sex‐ and diet‐dependent reductions in pathways that support brain mitochondrial energy metabolism.
Aberrant anabolic activity is critical to tumor biology; however, much remains to be learned about the regulators of protein anabolism in cancer and how this regulation may affect cancer pathophysiology. MicroRNA (miRNA), a family of small nucleotide regulatory molecules, may serve as a potential source of proteostatic regulation. Here, we examined the ability of two co-transcribed miRNA species, miR15a and miR16 (jointly described as miR15a/16) to regulate protein handling and pathophysiology in non-small cell lung cancer (NSCLC). We found that miR15a/16 regulates genes in numerous metabolic and pathological pathways, including those related to protein metabolism. Transfection of cellular models of NSCLC with miR15a/16 mimetics caused reductions in both cell growth and protein synthesis rates. These findings indicate that miR15a/16 acts as regulators of protein anabolism in NSCLC, serving as novel metabolic regulators and potential clinical therapeutic targets for malignant lung cancer.
ABSTRACT Osteoarthritis (OA)‐related meniscal degeneration involves complex interactions between oxidative stress and proteasomal dysfunction. However, the molecular drivers of regional meniscal vulnerability remain poorly defined. This study integrated multiple transcriptomic datasets from OA and control menisci to identify functional networks and hub genes by using weighted gene co‐expression network analysis. Human meniscal tissues from medial and lateral compartments were harvested during total knee arthroplasty and subjected to western blot analysis. In vitro assays on the basis of human chondrocytes were exposed to lipopolysaccharide or the proteasome inhibitor MG132 (carbobenzoxy‐l‐leucyl‐l‐leucyl‐l‐leucinal) to evaluate the stimulus‐specific regulation of identified network and hub genes. Weighted gene co‐expression network analysis revealed microsomal glutathione S‐transferase (MGST1) as the hub gene within a module enriched for ubiquitination and proteasome activity. Experimental validation in human meniscal tissues demonstrated pronounced upregulation of MGST1, ubiquitin‐conjugating enzyme E2 N (UBE2N), and proteasome activator complex subunit alpha (PSMA) in mechanically overloaded medial compartments compared to lateral regions. In vitro studies demonstrated stimulus‐specific modulation: lipopolysaccharide‐induced inflammatory stress upregulated MGST1, whereas proteasome inhibition via MG132 led to its downregulation. These findings highlight a dynamic interplay between redox adaptation and proteostasis, where chronic mechanical stress drives MGST1‐mediated antioxidant responses and compensatory ubiquitination. Together, these results suggest that joint tissues dynamically adapt to mechanical and inflammatory challenges by modulating oxidative stress defenses and protein quality control mechanisms, processes central to OA pathophysiology.
