Life Sciences

Melittin, a highly active natural antimicrobial peptide derived from honeybee venom, holds immense pharmacological potential against solid tumors. However, its specific anti-tumor efficacy and transcriptomic dynamics in cervical carcinoma remain to be systematically characterized. This study evaluated the anti-cancer properties of melittin on murine U14 cervical cancer cells following by transcriptomic investigation of the underlying mechanism. Phenotypic evaluations revealed that melittin potently inhibited U14 cell viability, while wound healing assays demonstrated a profound, dose-dependent suppression of cellular migration, culminating in near-complete migratory arrest at high concentrations. Furthermore, flow cytometry quantified a dramatic, dose-dependent surge predominantly in late apoptotic cell populations. These apoptotic events were structurally corroborated by scanning electron microscopy (SEM), which revealed severe plasma membrane perforation and morphological exhaustion. Enrichment analyses indicated that the physical membrane disruption inflicted by melittin translated into a severe metabolic crisis, marked by a global suppression of ribosomal biogenesis and mitochondrial oxidative phosphorylation. Furthermore, melittin profoundly repressed the Tie2-mediated angiogenic pathway (Etv2 down-regulation) while triggering the lethal hyperactivation of the AP-1 transcriptional stress complex (Jun, Fos, Fosb). Collectively, these findings elucidate the pharmacological networks underlying melittins cytotoxicity, providing solid molecular evidence for its development as a natural therapeutic agent against cervical cancer.

Mitochondrial dysfunction and oxidative stress represent two interconnected, primary causes for Diabetic Neuropathy (DN); however, the majority of currently available anti-diabetic therapies have focused on glucose control as opposed to neurodegenerative downstream effects. Corilagin, is an ellagitannin having high anti-oxidant properties; however, it has not been evaluated against hyperglycemia induced neuronal injury. The present study demonstrates the ability of Corilagin to protect against mitochondrial dysfunction via models of diabetic nephropathy and cerebral ischemia. High glucose (50 mM, 24 hr) was utilized to induce diabetes like conditions in the SH-SY5Y human neuroblastoma Cell Line. High glucose induced significant decreases in cell viability, increases in intracellular and mitochondrial reactive oxygen species, depletion of reduced glutathione reserves, induces apoptosis, and causes mitochondrial depolarization and fragmentation. Corilagin pre-treatment attenuated each of these high-glucose induced effects by protecting against mitochondrial membrane potential loss and maintaining mitochondrial network morphology while reducing apoptotic cell fraction relative to glucose alone. Additionally, these protective effects were accompanied by restoration of AMPK phosphorylation and up-regulation of SIRT1, PGC1 and TFAM, components that are part of the principal signaling pathway that regulates mitochondrial biogenesis; therefore, therefore, this pathway may contribute mechanistically to the cyto-protective effect of Corilagin. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=99 SRC="FIGDIR/small/740444v1_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@1be4a92org.highwire.dtl.DTLVardef@11d6e9org.highwire.dtl.DTLVardef@1346757org.highwire.dtl.DTLVardef@16c9f1e_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical abstractC_FLOATNO Proposed mechanism underlying the neuroprotective effects of Corilagin against high glucose-induced mitochondrial dysfunction.High glucose suppresses AMPK phosphorylation, leading to downregulation of the SIRT1-PGC-1-TFAM signaling axis, increased intracellular and mitochondrial reactive oxygen species (ROS), glutathione depletion, apoptosis, mitochondrial depolarization, and mitochondrial fragmentation. Corilagin pretreatment restores AMPK activation and the downstream SIRT1-PGC-1-TFAM pathway, thereby reducing oxidative stress, preserving intracellular glutathione, preventing apoptosis, maintaining mitochondrial membrane potential, and protecting mitochondrial network integrity. C_FIG

Histone deacetylases (HDACs) are pivotal epigenetic regulators that modulate diverse cellular pathways by removing acetyl groups from lysine residues on both histone and non-histone proteins. Histone deacetylase 11 (HDAC11), the sole member of class IV HDACs, exhibits both deacetylation and fatty acid deacylation activities. Accumulating evidence implicates HDAC11 as a key epigenetic regulator of fundamental cellular processes, including metabolism, immune responses, and tissue development. Dysregulation of HDAC11 activity has been associated with inflammatory diseases, metabolic disorders, neurodegenerative conditions, and cancer, highlighting its potential as a therapeutic target. Although several HDAC11-specific inhibitors have been identified, none have progressed to clinical development. In this study, we aimed to discover HDAC11-selective inhibitors by integrating in silico and in vitro validation approaches. Homology modelling of the HDAC11 structure was conducted, followed by model validation, structure-based virtual screening, molecular dynamics (MD) simulations, and binding free energy calculations. We identified and validated three lead compounds and their intermediates using biochemical and cell-based assays. Fluorescence-based and HPLC-based enzymatic assays demonstrated potent inhibition of both the deacetylase and deacylase activities of HDAC11, with Inhibitor 6 and Inhibitor 3 exhibiting the strongest effects among the six compounds tested. Further, a decrease in lipid accumulation, reduced stability of the HDAC11 substrate SHMT2, as determined by immunoblot analysis and decreased cell viability, as assessed by MTT assay, confirmed HDAC11 inhibition in cellular models. The study shows that new HDAC11 inhibitors significantly reduce the viability of breast cancer cells and induce apoptosis; inhibitor 6, in particular, showed high potency, similar to the reference compound SIS-17. Flow cytometry showed that treated MDA-MB-231 cells exhibited cell-cycle arrest and increased apoptosis, a finding further confirmed by Annexin V/PI staining. Molecular analysis showed that BAX increased while BCL2 decreased, indicating that apoptotic pathways were activated in novel compound-treated MDA-MB-231 cells. The results suggest that inhibiting HDAC11 is an effective way to induce cancer cell death and provide a basis for further assessment of these compounds as potential treatments for breast cancer. Collectively, this study identifies novel zinc-chelating HDAC11 inhibitors containing a nitro-sp2 group, providing promising candidates for further therapeutic development.

Subarachnoid hemorrhage (SAH) resulted from intracranial aneurysm (IA) rupture is an especially severe form of stroke. Endothelial dysfunction represents the initiating event of IA pathogenesis. Understanding the role of endothelial cells (ECs) underlying formation of IAs is helpful to seek for pharmaceutical treatment strategy. Based on single-cell RNA sequencing, proteomics, and metabolic analysis, we discovered a change in cell population in IA samples, majorly in ECs and macrophages (MPs). Abnormal ECs exhibit senescence and death in IA samples, which is absent in the control arterial samples. Cross-analysis of multi-omics revealed that CALM1, a calcium detector involved in mechanotransduction, is downregulated in the abnormal ECs. CALM1 knockdown leads to senescence and inhibits the proliferation and maturation of ECs under turbulent flow. Through high-throughput virtual screening, this work identified compound ZC04329651 as a potent CALM1 activator in maintaining the stability of endothelial cell junctions and attenuating cellular senescence. Thus, our findings showed compound ZC04329651 up-regulate the expression of CALM1 to restore ECs, which maybe a promising pharmacological treatment strategy for IAs.

Background Intrauterine inflammation is associated with increased lifetime cardiovascular disease (CVD) risk in offspring, yet long-term molecular mechanisms remain underexplored. We investigated the lasting cardiometabolic, hemodynamic, and vascular effects of prenatal inflammatory exposure in adult offspring using a mouse model. Methods On day 15 of gestation, pregnant CD-1 mice were randomized to receive intraperitoneal (IP) injections of either lipopolysaccharide (LPS group) or saline (SAL group). Blood samples were collected before and after injection to measure IL-6 concentrations. Blood pressure (BP) in offspring was measured at 6-8 months of age using the tail-cuff method, and tissues were collected. Protein was extracted from the brain cortex to measure cytokines utilizing the xMAP Luminex assay. The C-reactive protein (CRP) was measured in serum. Gene expression of IL-6, NOS3, NFkB, AT-1, and Trp53 was determined by real time RT-qPCR in the aorta. The experimental unit was a mother. Results were analyzed using the appropriate statistical methods. Results Maternal LPS administration induced significant acute systemic inflammation (P<0.05 for IL-6). At 6-8 months after delivery, maternal circulating CRP was significantly higher in LPS mice as compared to SAL (P=0.02). In offspring at 6-8 months of age, systolic BP was significantly elevated in LPS groups compared with SAL animals (P=0.04). LPS-exposed offspring exhibited higher total body and visceral adipose tissue weights. Aortic expression of the angiotensin II type 1 receptor (AT1) and NFkB were significantly upregulated (P=0.04 and P=0.03, respectively). Cortices of LPS offspring displayed a pro-inflammatory profile with increased total Inflammatory score, elevated pro-inflammatory IL-1{beta}, and reduced anti-inflammatory IL-4 and IL-10. Furthermore, protective Trp53 gene expression was significantly downregulated in both the aorta (P=0.01) and heart (P=0.04). Conclusion A single mid-gestation inflammatory insult programs long-term cardio-inflammatory perturbations in offspring and results in long-term systemic inflammation in mothers. Prenatal LPS exposure induces persistent visceral obesity, hypertension, vascular RAS upregulation, central neuroinflammation, and cardiac/vascular Trp53 suppression in adult offspring, offering a mechanistic link between prenatal inflammation and offspring CVD risk.

AimThe aim of the present study was to determine whether pharmacological inhibition of aldosterone synthesis during the stress-hyporesponsive period (SHRP) affects behaviour and adrenocortical stress responsiveness later in development and whether these effects differ between males and females. MethodsNewborn Wistar rat pups (males n=40, females n=40) were treated with aldosterone synthase inhibitor FAD286 (30 mg/kg per day, orally) or vehicle from PND3 to PND9. To verify the pharmacodynamic action of FAD286, serum and adrenal glands from 10-day-old pups were analysed. The remaining pups were weaned on PND21 and underwent open-field (PND23), elevated plus-maze (PND29) and salt-preference testing. At PND46, half of each group was exposed to restraint stress for 120 min. ResultsIn 10-day-old pups, treatment with FAD286 resulted in increased gene expression of CYP11B2 (aldosterone synthase) and CYP11B1 (11-beta-hydroxylase) in the adrenal glands, increased serum levels of corticosterone, and decreased concentrations of serum aldosterone. FAD286 did not modify the general locomotor activity assessed in juvenile rats. Inhibition of aldosterone synthase by FAD286 resulted in altered anxiety-like behaviour in a sex-dependent manner. Postnatal FAD286 treatment led to increased anxiety-like behaviour in female, but not male rats. During adolescence, early FAD286 treatment increased overall aldosterone concentrations without altering the aldosterone response to restraint. Basal corticosterone concentrations were unchanged, whereas the response to restraint was enhanced. ConclusionsThe present study demonstrates that transient inhibition of aldosterone synthesis during the SHRP led to alterations in anxiety-related behaviour and adrenocortical regulation later in development, with some behavioural effects being sex-dependent.

RationaleOxidative stress, resulting from an imbalance between the production of reactive oxygen species (ROS) and antioxidant defence mechanisms, disrupts cellular redox homeostasis and contributes to damage of biomolecules, including nucleic acids, proteins and lipids. Such disturbances can impair intracellular signalling pathways and have been implicated in the pathophysiology of female reproductive disorders. Nitroblue Tetrazolium (NBT) is a distinctive dye that assesses cellular redox activity in neutrophils through the detection of superoxide anion (O2-). However, the relationship between neutrophil-derived oxidative stress and women infertility remains to be fully elucidated. Aim of the study: We aimed to investigate the role of neutrophil oxidative stress in women infertility using a novel Nitroblue Tetrazolium (NBT) method developed in our laboratory. Materials and methodsBlood and serum specimens were obtained from a total of 100 Libyan women, comprising healthy fertile women (n = 21; controls) and infertile women (n = 79). Superoxide anion (O2-) generation in neutrophils was assessed using the novel NBT method, while malondialdehyde (MDA), a marker of lipid peroxidation, was determined using the thiobarbituric acid reactive substances (TBARS) assay. ResultsA significant increase in both NBT-reactivity levels and the percentage of NBT-positive neutrophils was observed in infertile women compared with healthy fertile controls (P < 0.0001). In addition, MDA levels were significantly higher in infertile women than in the control group, indicating enhanced lipid peroxidation. MDA levels were positively correlated with NBT-reactivity levels (r = 0.410, P = 0.0001) and the percentage of NBT-positive neutrophils (r = 0.21, P = 0.047). A significant positive correlation was also observed between NBT-reactivity levels and the percentage of NBT-positive neutrophils (r = 0.510, P = 0.0001), demonstrating a close association between neutrophil oxidative activity and lipid peroxidation in women infertility. ConclusionThe present study demonstrates the utility of a novel NBT method for detecting reactive oxygen species (ROS), a marker of oxidative stress, in neutrophils from both blood and serum specimens. The findings demonstrate increased neutrophil-derived oxidative stress in infertile women and suggest that this method may have potential as a diagnostic tool for the assessment of women infertility. ImpactThis study provides evidence that neutrophil-derived oxidative stress is significantly increased in women infertility and demonstrates the application of a novel Nitroblue Tetrazolium (NBT) assay for assessing oxidative stress in both blood and serum specimens. The significant associations between NBT-reactivity, NBT-positive neutrophils and malondialdehyde levels provide further insight into the contribution of oxidative stress to female reproductive pathophysiology. These findings support the potential utility of the novel NBT method as a simple and reliable diagnostic approach for evaluating oxidative stress in women infertility and provide a foundation for future studies investigating oxidative stress biomarkers in reproductive medicine.

Background: Early detection of preeclampsia with noninvasive and reliable biomarkers is the foremost step for minimizing adverse effects during pregnancy. However, none maternal serum analyte evaluated so far is sufficiently accurate to allow recommending their routine use. Methods: Microarray was used to first identify differentially expressed LncRNA and mRNA. Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to determine bio-functions and signaling pathways. Quantitative real-time polymerase chain reaction was used to validate the results of microarray. Finally, the lncRNA-mRNA co-expression network was constructed to find the interaction of genes. Result: A total of 1476 differentially expressed lncRNAs and 594 mRNAs were identified. Both RNA-seq and RT-qPCR showed the expression of T241171,T338586, uc002ywy.3 was up-regulated, and the expression of ENST00000524858, T131416, T357032, uc.335+ was down regulated in the plasma of patients with preeclampsia. Pathway analysis showed the targeted mRNAs were enriched in apoptosis, sulfur metabolism, starch and sucrose metabolism. ceRNA network found that LncRNA-uc002ywy.3 might be the upstream regulator of miRNA-4498. LncRNA-uc002ywy.3/miRNA-4498 was predicted to interplay with genes involved in programmed cell death-1and its ligand (PD-1/PD-L1) pathway by KEGG analysis. Conclusion s PD-1/PD-L1 signaling pathway may be involved in the development of preeclampsia. The dysregulated LncRNA-uc002ywy.3/miRNA-4498 shed light on a new layer involved in the regulatory network of preeclampsia.

Background: Metabolic Syndrome (MS) is an escalating non-communicable disease (NCD) and public health crisis heavily driven by modern high-calorie dietary transitions. MS is closely linked to catastrophic cardiovascular dysfunction (CVD). Conventional CVD medications are frequently constrained by high financial costs, system-wide adverse effects, and poor population-level accessibility in lower-middle-income countries (LMICs). This underscores an urgent public health need to evaluate safe, scalable, and low-cost dietary nutraceuticals for primary prevention. This study evaluated the cardioprotective and preventative effects of Beta-caryophyllene (BCP)--an affordable, widely accessible, and GRAS-designated dietary cannabinoid--in a lifestyle-mimicking rodent model of MS. Methods: Seventy-two male Wistar rats (10 weeks old) were divided into six groups (n=12/group). Groups I and II served as controls given Normal Diet (ND) alone and ND+{beta}-caryophyllene (50 mg/kg p.o.), respectively. Metabolic Syndrome was induced in Groups III-VI using a High-Fat High-Fructose Diet (HF/HFD) for 16 weeks. For the subsequent 4 weeks, Group III continued on HF/HFD, Group IV received HF/HFD + BCP (50 mg/kg p.o.), Group V underwent dietary withdrawal (returned to a normal diet [ND]), and Group VI received ND + BCP. Conscious hemodynamic variables and electrocardiograms (ECG) were recorded via telemetry. Plasma apolipoprotein-B (Apo-B) and C-reactive protein (CRP) were quantified using ELISA. Lipid profiles, oxidative stress parameters, and organ histopathology were evaluated using standard biochemical and staining methods, with data analysed using ANOVA at = 0.05. Results: In Group II, BCP decreased SBP (145.00{+/-}12.13 vs 120.30{+/-}2.15 mmHg), DBP (105.70{+/-}10.80 vs 94.00{+/-}17.20 mmHg), heart rate (448.00{+/-}11.90 vs 280.00{+/-}18.70 beats/min), and PR-interval (49.50{+/-}3.53 vs 45.00{+/-}12.70 ms) compared with Group I. Group IV significantly decreased SBP (129.00{+/-}1.28 vs 125.5{+/-}11.1 mmHg) and heart rate (380.00{+/-}16.20 vs 351.00{+/-}3.00 beats/min) compared with Group III. Furthermore, BCP significantly increased HDL-cholesterol (10.80{+/-}1.70 vs 15.40{+/-}1.00 mg/dl), SOD (0.35{+/-}0.07 vs 0.76{+/-}0.30 U/mg protein), and GSH (1.64{+/-}0.02 vs 3.60{+/-}0.13 mM) in Group IV compared with Group III. Conversely, BCP significantly decreased total cholesterol (166{+/-}31.00 vs 113{+/-}18.50 mg/dl), Apo-B (26.70{+/-}1.68 vs 19.00{+/-}1.30 mg/dl), leptin (5.10{+/-}0.03 vs 2.18{+/-}0.01 ng/ml), NO (6.30{+/-}0.40 vs 4.30{+/-}2.61 M), and malondialdehyde (11.80{+/-}4.10 vs 9.90{+/-}2.30 M) in Group IV. BCP significantly decreased triglycerides (64.90{+/-}1.30 vs 55.90{+/-}1.30 mg/dl) in Group VI compared with Group IV, while lowering CRP across all groups. Distortion of the cytoarchitecture of the heart, liver, and pancreas observed in Group III was completely reversed by BCP treatment in Group IV. Conclusion: Beta-caryophyllene effectively ameliorates diet-induced metabolic syndrome and associated cardiovascular dysfunction via potent anti-atherogenic, anti-inflammatory, and antioxidant mechanisms. These findings suggest that integrating affordable, culturally accepted, BCP-rich indigenous functional ingredients into regional nutrition frameworks represents a highly scalable, low-cost public health strategy for global NCD prevention. Keywords: Cardiovascular dysfunction, Metabolic Syndrome, Beta-caryophyllene, High-fat-high-fructose diet, Public Health Nutrition, NCD Prevention.

Environmental exposure to endocrine disrupting chemicals, like bisphenol-A (BPA), can impart detrimental effects on developing feto-placental unit, during pregnancy. Placenta remains a central player maintaining this feto-placental homeostasis for sustenance of a healthy pregnancy. Thus, the bisphenol-A mediated endocrine disruption affects the healthy functioning of placenta by altering key processes, such as tissue remodelling, angiogenesis, and metabolism. However, the underlying mechanism of BPA-altered ECM remodelling remains elusive. Therefore, in this study we investigated the BPA mediated changes in placental tissue remodelling using a bisphenol-A exposed murine model during pregnancy. The results reveal that, the phenotypic changes in feto-placental interface correlates with perturbed placental proteome in response to BPA. Further investigation highlights a S100a10-Annexin A2 axis mediated upregulation of tissue plasminogen activator (tPA), which drives altered extracellular matrix (ECM) degradation in placental decidua. This culminates into functional dysregulation in feto-placental axis, leading to reduced size of fetus and placenta. Therefore, this study provides novel insights of a S100a10-Annexin A2 axis associated mechanism for alteration of ECM remodelling in placental decidua due to BPA exposure, which may lead to toxicity related adverse pregnancy outcome.

Introduction: Preterm birth is often associated with fetal membrane (FM) inflammation. Opioid-use disorder during pregnancy is a major public health issue as all opioids carry a risk for preterm birth. While maintenance therapy with methadone improves maternal outcomes, it also elevates the risk of preterm birth. We previously reported that the opioid, buprenorphine, induced human FM sterile inflammation. The goal of this study was to investigate if methadone also induced FM inflammation and to determine the mechanisms involved. Methods: Human FM explants were treated with or without methadone in the presence or absence of inhibitors to TLR4/MD2, TLR7, TLR8, NLRP3, the -opioid receptor, p65 NF{kappa}B, p38 MAPK, ERK or JNK. Supernatants were measured for cytokines/chemokines by ELISA. Tissues were analyzed by Western blot for NF{kappa}B and MAPK activity, and by RT-qPCR for the TLR7/8-activating miRs: miR-21a, miR-29a, miR-146a-3p, Let-7b. Pregnant C57BL/6 mice were injected with PBS or methadone on E15.5 and 6hrs later FMs were analyzed by RT-qPCR. Results: Methadone elevated human FM explant secretion of IL-1{beta}, TNF, MIP-1, GM-CSF, and PGE2, and expression of miR-21a. In vivo, methadone elevated mouse FM expression of MIP1A, KC, and COX2. TLR4/MD2 inhibition had no effect on human FM inflammation in response to methadone. NLRP3 inhibition reduced methadone-induced FM IL-1{beta}, TLR7 inhibition reduced methadone-induced FM TNF, and inhibition of either TLR8 or the -opioid receptor reduced methadone-induced secretion of TNF, MIP-1, GM-CSF, and PGE2, but not IL-1{beta} Methadone increased human FM activity of p65 NF{kappa}B, p38 MAPK, JNK, and ERK and inhibition of these signaling pathways collectively reduced methadone-triggered FM inflammation. Finally, naloxone reduced most of the methadone-induced FM inflammatory response. Conclusion: Methadone induces sterile inflammation in human and mouse FMs. In human FMs this inflammation is mediated through activation of innate immune TLR7, TLR8, and NLRP3; the -opioid receptor; and NF{kappa}B/MAPK signaling. These findings provide further insight into the mechanisms underlying the association between opioid use during pregnancy and preterm birth.

INTRODUCTIONSex differences in mental health emerge during adolescence, a period marked by the onset and the establishment of menstrual cycle. However, is rarely examined how menstrual cycle regularity, a marker of hormonal function, modulates mental health. OBJECTIVEto analyse sex differences in mental health symptoms among adolescents considering the menstrual cycle regularity and sleep. METHODSA three-group design (female students with regular cycles/FR, n=77; with irregular cycles/FI, n=59; and male students/M, n=76) in a sample of Brazilian high-school adolescents (n=212; 14-18 years) enrolled in morning and full-time classes was used to test the hypothesis that mental health symptoms follow a graded pattern across these groups. RESULTSMean DASS-21 scores across all groups fell at or above the Mild severity threshold for mental health subscales. GLMs confirmed a monotonic gradient increase in group order (M[-&gt;]FR[-&gt;]FI) which was associated with higher scores on all outcomes (stress {beta}/step=4.33, p<.001; anxiety {beta}/step=4.08, p<.001; and depression {beta}/step=2.34, p=.010; model R{superscript 2}=.16, .13, .08 respectively). However, no differences were observed in sleep duration, social jetlag, chronotype, sleep quality, or sleep-debt. Then, a secondary analysis assessed sex-specific associations between socioeconomic status (SES) and mental health; higher SES was inversely related to stress, anxiety, and depression, being protective only in males (stress Males {beta}=-2.68, p=.011/Females {beta}=0.46, p=.614). CONCLUSIONThese findings support the reframing of menstrual irregularity not only as a reproductive health concern but also as a biological determinant of mental health risk in female adolescents, a vulnerability that sleep disruption and socioeconomic resources do not adequately explain.

Lymphedema is a chronic disease characterized by impaired lymph drainage and accumulation of protein-rich interstitial fluid, which progresses to develop irreversible fibrosis. Importantly, effective therapies and treatments are lacking to alleviate and mitigate the disease. The pathological inflammation and fibrogenesis underlying lymphedema prompted us to evaluate a preclinical medicine IVA337, a pan-peroxisome proliferator-activated receptor (PPAR) agonist that improves liver fibrosis in patients with metabolic dysfunction-associated steatohepatitis (MASH) by activating three PPAR isoforms (, {beta}/{delta}, {lambda}), which play critical roles in lipid metabolism, anti-inflammation responses, and anti-fibrogenesis. Here, we investigate the therapeutic effects of IVA337 during the early stage of surgery-induced secondary lymphedema in mice and explored the underlying mechanisms. IVA337 administration alleviated lymphedema progression, improved lymphatic drainage, reduced dermal thickness, and resolved lymphatic vessel dilation. Mechanistically, IVA337 suppressed the TGF{beta}/SMAD2/3 signaling pathway, reduced immune cells infiltration, and improves lymphatic vessels integrity. In human dermal lymphatic endothelial cells (HDLECs), IVA337 attenuated TGF{beta} induced SMAD2/3 phosphorylation and preserved the expression of cell junction Claudin5, reduced VE-Cadherin-stained cell-cell gaps. Collectively, our findings demonstrate that IVA337 protects against early stage lymphedema by inhibiting TGF{beta}/SMAD2/3-mediated inflammatory and fibrotic responses. This study provides a potential therapeutic strategy to improve lymphatic function during the early phase of lymphedema and prevent progressive fibrosis in patients with lymphedema and related disorders.

Neurotensin receptor 1 (NTSR1) activation suppresses feeding and promotes weight loss but is limited by adverse effects associated with Gq signaling. SBI-553 is a {beta}-arrestin-biased allosteric modulator of NTSR1 that avoids these effects. We evaluated the effects of SBI-553 (5 or 12 mg/kg, i.p.) on food intake and body weight in lean and diet-induced obese mice. Neither dose altered metabolic parameters, locomotor activity, or wheel running. SBI-553 did not affect ad libitum feeding in lean mice; however, both doses acutely reduced high-fat diet intake in obese mice. While 5 mg/kg had no effect on hunger-induced feeding, 12 mg/kg suppressed refeeding in both chow- and high-fat-fed mice of both sexes and reduced weight regain in obese mice. These findings identify SBI-553 as a potential strategy for reducing food intake and supporting weight loss.

Plant-derived immunomodulators are of immense therapeutic interest, of which sesquiterpene lactones (SLs) are a promising source. Artemisinin and Parthenolide are two structurally related SLs that are known to inhibit NF-{kappa}B. However, whether this mechanism produces comparable immunomodulatory outcomes has not been directly examined. In this study, we compared the effects of Artemisinin and Parthenolide across different in vitro and in vivo inflammatory contexts: T-cell activation, macrophage activation, and Salmonella Typhimurium infection. Parthenolide inhibited mouse T-cell activation more effectively than Artemisinin in terms of cell cycling, IL-2 production, and induction of activation markers, CD69 and CD44. In addition, Parthenolide suppressed production of LPS-induced nitrite and pro-inflammatory cytokines in primary thioglycolate (TG)-elicited macrophages as well as RAW 264.7 cells. It also reduced ROS production across all tested conditions. In contrast, Artemisinin exhibited comparatively modest effects, although it increased IL-6 in LPS-activated RAW 264.7 cells. Neither compound reduced bacterial burden in an in vitro model of S. Typhimurium infection; however, Parthenolide, but not Artemisinin, lowered TNF- amounts. Together, these findings demonstrate that Parthenolide is a broader and more consistent immunomodulator than Artemisinin. These observations led us to investigate the effects of Parthenolide in an in vivo model of mice orally fed with S. Typhimurium. Parthenolide-treated mice showed higher survival accompanied with lower sera amounts of IL-6 and TNF-, indicating that its protective effects operate by limiting host immunopathology. The implications of these observations on the use of compounds that lower host inflammatory responses during infections are discussed.

Obesity is a major public health challenge of the 21st century, particularly in low- and middle-income populations. Adipogenesis plays a central role in the development of obesity and associated metabolic disorders, as it determines adipocyte number, size, and function. The 14-3-3 protein family comprises seven paralogs in mammals that regulate multiple cellular processes, yet their specific roles during adipogenesis remain poorly understood. In this study, we characterized the expression profiles of 14-3-3 paralogs during the early and late stages of adipogenic differentiation using quantitative PCR under standard adipogenic differentiation medium and modified drug-supplemented conditions. We found that the expression of specific paralogs is strongly influenced by the composition of the differentiation medium. The absence of insulin led to an early increase in Ywhaz, which could not be maintained during late adipogenesis and was associated with impaired adipogenic differentiation. In contrast, stimulation with incretins in combination with insulin induced late expression of Ywhag and Ywhab paralogs and promoted the formation of a greater number of smaller lipid droplets. These findings indicate that individual 14-3-3 paralogs exert distinct and context-dependent effects on adipogenesis, highlighting their potential roles as modulators of adipocyte differentiation and metabolic function.

REV-ERB{beta} is a transcriptional repressor of nuclear receptors that regulates the circadian rhythm and plays an important role in regulation of the proliferation, differentiation, and maturation of neurons. Dysregulation of the circadian rhythm has been associated with neuropsychiatric disorders, and activation of REV-ERBs can induce anxiolytic behavior in mice. Furthermore, hippocampal neurogenesis is important in the effects of antidepressants. However, the role of REV-ERB{beta} in adult hippocampal neurogenesis in vivo at the single-cell level is not known. In this study, protein localization of REV-ERB{beta} in the subgranular zone of the hippocampal dentate gyrus (DG) was mainly shown in NeuN-positive neurons, and the effect of expressing a dominant negative form of REV-ERB{beta} lacking the C-terminal region on newborn neurons in the hippocampal DG of adult mice was examined to investigate the role of REV-ERB{beta} in neurogenesis. A retroviral vector containing the dominant negative REV-ERB{beta} or a control vector was injected into the mouse DG. At 4 weeks after injection, the morphology of dendrites and dendritic spines of newborn neurons labeled by the virus was examined. Expression of the dominant negative form of REV-ERB{beta} inhibited dendrite outgrowth and branching and decreased dendritic spine formation in newborn neurons in the adult mouse hippocampal DG. This study revealed a new role for REV-ERB&{beta} in adult hippocampal neurogenesis at the single cell level, and the results will provide insight into neurogenesis in the adult brain and its relationship with psychiatric disorders.

BackgroundCardiac fibrosis drives adverse myocardial remodelling through persistent fibroblast activation, ECM deposition, and impaired cardiac function. Current therapies offer limited protection against cardiac fibrosis progression. Elafibranor is a dual PPAR-/{delta} agonist approved for the treatment of liver disease. However, its effects in human models of cardiac fibrosis remain insufficiently explored. MethodsElafibranor was evaluated in complementary human in vitro TGF-{beta}1-induced cardiac fibrosis models: 2D primary fibroblasts, 3D fibroblast spheroids, spontaneously contracting 3D cardiac microtissues, and hiPSC-derived cardiomyocytes. Viability, apoptosis, fibroblast activation, ECM remodelling, mitochondrial respiration, nucleotide and NAD pools, calcium handling, contractility, and transcriptomic profiles were assessed. ResultsAt non-cytotoxic concentrations, elafibranor attenuated TGF-{beta}1-driven cardiac fibrosis responses. In 2D cardiac fibroblasts, it reduced myofibroblast differentiation, procollagen 11 secretion, and partially restored mitochondrial respiratory capacity. In 3D spheroids, it preserved viability, attenuated caspase-3/7 activation, and suppressed procollagen 11 release. In cardiac microtissues, elafibranor reduced ECM accumulation, shifted transcriptomic profiles toward redox-metabolic/cytoprotective pathways, altered adenine nucleotide and NAD pools, and partially recovered contraction parameters. In hiPSC-derived cardiomyocytes, elafibranor modulated calcium handling, contractility, and mitochondrial respiration. ConclusionsElafibranor mitigates TGF-{beta}1-driven cardiac fibrosis by suppressing fibroblast activation and ECM remodelling while promoting adaptive metabolic, redox, and bioenergetic responses, supporting balanced PPAR-/{delta} activation as a potential therapeutic strategy for cardiac fibrosis. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/745425v1_ufig1.gif" ALT="Figure 1"> View larger version (54K): org.highwire.dtl.DTLVardef@1cbd94eorg.highwire.dtl.DTLVardef@27a44borg.highwire.dtl.DTLVardef@9354baorg.highwire.dtl.DTLVardef@9f9946_HPS_FORMAT_FIGEXP M_FIG C_FIG

Objectives: The bile acid nuclear receptor Farnesoid X Receptor (FXR, NR1H4) is a major regulator of metabolism and energy homeostasis in peripheral organs. It modulates bile acid, glucose, and lipid metabolism, as well as fat mass and body weight. However, FXR is also expressed in the brain, particularly in the hypothalamus, a key center for the regulation of energy homeostasis. Although one study has demonstrated a role for brain FXR activation in energy balance, its specific hypothalamic role is still unknown. Here, we examined the role of FXR in the mediobasal hypothalamus in the regulation of energy balance. Methods: We used a genetic approach combined with metabolic phenotyping to determine the effect of FXR invalidation in the mediobasal hypothalamus on metabolic parameters involved in the central regulation of energy homeostasis. Results: Our results demonstrate that hypothalamic FXR deficiency induces a positive energy balance, resulting in a reduction in energy expenditure due to alterations in glucose metabolism accompanied by structural changes in white adipose tissues. Conclusion: This study uncovers a previously unrecognized role for hypothalamic FXR in the central homeostatic control of energy balance, providing new insights into its contribution to peripheral glucose metabolism and adipose tissue structural remodeling.

Kaposi's sarcoma (KS) is an angioproliferative tumor associated with infection by human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma-associated herpesvirus (KSHV). Among the viral genes involved in KS pathogenesis, the viral G protein-coupled receptor (vGPCR) plays a key role in oncogenic transformation and angiogenesis, and its sustained activity contributes to tumor development and maintenance. Quercetin (QUE), a naturally occurring flavonoid with well-recognized anticancer properties, has been shown to reduce cell viability and proliferation and to induce apoptosis in vGPCR-transformed endothelial cells. This study further investigated the mechanisms underlying the antineoplastic effects of QUE in vGPCR cells, an in vitro model of Kaposi's sarcoma. Initially, QUE promoted the activation of MAPK signaling pathways in vGPCR cells, increasing the phosphorylation levels of ERK1/2, JNK1/2, and p38, without affecting AKT phosphorylation. In parallel, QUE increased intracellular oxidant levels, followed by lipid peroxidation and DNA damage, supporting a link between oxidative imbalance and genotoxicity. However, increased total glutathione content, and unaltered Gpx4 mRNA expression, suggest the induction of oxidative stress and compensatory antioxidant responses. Furthermore, increased intracellular oxidant levels were associated with MAPK signaling activation, particularly JNK1/2, whereas ERK1/2 and p38 activation may reflect a compensatory response that is insufficient to overcome QUE-induced cytotoxicity. Taken together, these findings indicate that QUE exerts antiproliferative effects in vGPCR cells through a mechanism involving oxidative stress, MAPK activation, and DNA damage, supporting its potential relevance in the study of therapeutic strategies for virus-associated malignancies.