Polycystic ovary syndrome (PCOS) is a prevalent endocrine abnormality affecting the women of child bearing age. PCOS markedly raises the chances of infertility, cardiovascular disease, and increased blood pressure. Ocimum tenuiflorum is known for its diverse roles and is used in Ayurveda to address a variety of pathological conditions. This study was done to scrutinize the role of inflammatory genes such as iNOS, COX-2, IL-1β, and TNF-α in letrozole-induced PCOS in female Wistar rats and to assess the therapeutic potential of Ocimum tenuiflorum extracts (OTM and OTEA) in restoring biochemical, histological, and molecular parameters through western blot and RT-PCR analyses. In this research, a total of sixty albino female Wistar rats were employed. Eighteen rats were designated for evaluating the toxicity of the plant extracts, and the remaining 42 were distributed randomly across seven groups. PCOS was induced by administering letrozole at a dosage of 1 mg/kg body weight, which was formed in 0.5% CMC solution, for a duration of 21 days. After induction (21 days), the rats received oral treatment with methanol (OTM) and ethyl acetate (OTEA) extracts of Ocimum tenuiflorum at the dose of 50 and 100 mg/kg body weight, while metformin (25 mg/kg body weight) was used as the positive control. The plant material underwent Soxhlet extraction, resulting in five separate extracts: hexane, ethyl acetate, ethanol, methanol, and aqueous. The in vitro anti-inflammatory efficacy of these extracts was assessed using protein denaturation, proteinase inhibition, erythrocyte membrane stabilization, and nitric oxide scavenging assays. An initial toxicity assessment was conducted to determine the safety of these extracts. Subsequently, in vivo evaluation included ovarian histology, serum biochemistry, and molecular analysis. Serum levels of sex hormones (testosterone and estradiol) were measured and the expression of inflammatory markers (iNOS, COX-2, TNF-α, and IL-1β) was examined using RT-PCR and western blotting. GC-MS analysis was performed to identify the compounds responsible for the pharmacological results. At 600 μg/mL concentration, the extracts of methanol and ethyl acetate showed notable anti-inflammatory effects, with inhibition of protein denaturation at 88.22 ± 1.52% and 82.51 ± 1.74% (P < 0.001), proteinase activity at 93.43 ± 1.6% and 89.03 ± 1.46% (P < 0.0001), erythrocyte membrane haemolysis at 89.61 ± 0.93% and 88.94 ± 0.92% (P < 0.0001), and nitric oxide activity at 88.34 ± 1.43% and 90.14 ± 1.2% (P < 0.0001), respectively. In vivo experiments using Wistar rats demonstrated that the oral administration of potent extracts did not produce any toxic effects. Post-treatment with OTM and OTEA extracts in PCOS-induced rats resulted in decreased serum testosterone and estrogen levels. A notable reduction in apparent follicular cysts was seen in the ovaries, along with a marked down-regulation in the expression levels of mentioned inflammatory genes suggesting potential anti-inflammatory and PCOS-modulating effects. Finally, GC-MS analysis confirmed the active compounds with robust anti-inflammatory efficacy. Our study showed that Ocimum tenuiflorum has the potential to attenuate PCOS by restoring hormonal levels and exerting anti-inflammatory effects.
Focal cortical dysplasia (FCD) is a leading cause of drug-resistant epilepsy, whereas its molecular and cellular mechanisms remain poorly understood. This study aimed to characterize the cellular heterogeneity of FCD and investigate the function of ferroptosis in FCD pathogenesis. Single-nucleus RNA sequencing was carried out on epileptogenic cortical tissues from 18 patients with FCD and 6 perilesional control samples with normal histology. Data were analysed using uniform manifold approximation and projection for dimensionality reduction and visualization. Differentially expressed genes (DEGs) were identified and subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses. UCell scoring and gene set enrichment analysis (GSEA) were applied to assess pathway activity. Expression levels of ferroptosis-related genes (FRGs) were validated by immunofluorescence, and biochemical assays quantified the levels of superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA) and lipid peroxides (LPO). A total of 170 747 nuclei were profiled, resolving five major cell types, including inhibitory neurons, excitatory neurons, astrocytes, microglia and oligodendrocytes. DEGs across these populations were significantly enriched in ferroptosis and oxidative stress-associated pathways. UCell and GSEA highlighted remarkable alterations in ferroptosis, apoptosis and oxidative stress, particularly in inhibitory neurons and astrocytes. Immunofluorescence confirmed upregulation of key FRGs, including ferritin light chain, ferritin heavy chain 1, poly rC binding protein 1, microtubule-associated protein 1 light chain 3B and prion protein-encoding gene, in FCD tissues. Concordantly, biochemical assays demonstrated reduced SOD and GSH levels, alongside elevated MDA and LPO levels, confirming the transcriptional and histological findings. The results indicated that ferroptosis may play a notable role or act as a concurrent mechanism in the pathogenesis of FCD, potentially contributing to the neuronal and glial dysfunction and epileptogenesis. Integrating transcriptomic, histological and biochemical data, this study demonstrated that targeting ferroptosis-related pathways may hold promise as a potential therapeutic strategy for FCD, providing new insights into the molecular mechanisms underlying this condition. This study pioneers the first single-nucleus transcriptomic atlas for Focal Cortical Dysplasia (FCD) types I and II, deciphering the cellular heterogeneity across five major brain cell types within the epileptogenic cortex. Through integrated multi-omics analysis, it reveals for the first time a significant association between the ferroptosis pathway and FCD pathogenesis. We identify and validate ferroptosis-related genes (e.g., FTH1, FTL, PCBP1) as potential biomarkers and therapeutic targets, supported by congruent biochemical evidence of oxidative stress in this drug-resistant epilepsy.
Ulcerative colitis (UC) is a global inflammatory bowel disease (IBD) and is a chronic mucosal inflammation of the large intestine. UC is accompanied by the increment in the production and release of pro-inflammatory mediators. Due to the immunomodulatory potentials of Berberine (BBN), the present study aimed at examining its anti-ulcerogenic activity against experimentally induced ulcerative colitis (UC), by intrarectal instillation of 1 ml of 3% acetic acid (AA). Thirty adults female Wistar rats were divided into three groups: (1) Negative control, (2) AA-induced UC rats (intrarectal), (3) Treated AA-induced UC + BBN (50 mg/kg/day; orally). Biochemical, molecular, histopathological, and immunohistochemical investigations were conducted. Intrarectal administration of AA provoked several macroscopic and microscopic alterations in the colons of UC-induced rats, increased the colonic lipid peroxidation, upregulated the expression of nuclear factor kappa B (NF-κB), caspase-3, and interferon gamma (IFN-γ), increased levels of colonic inflammatory tumor necrosis factor-alpha (TNF-α), Interleukin-1 beta (IL-1β), and prostaglandin E 2 (PGE-2), and downregulated the immunoexpression of nuclear factor erythroid 2-related factor 2 (Nrf-2). In contrast, treatment of UC-rats with BBN exhibited curative activities manifested by downregulating the expression of NF-κB and caspase-3, reducing the colonic contents of malondialdehyde (MDA), TNF-α, IL-1β, and PGE-2; and activating Nrf-2 immunoexpression. This study evidenced the anti-ulcerative and colo-therapeutic potentials of BBN that might be ascribed to its anti-lipid peroxidation, anti-apoptotic, and anti-inflammatory activities.
Early-onset colorectal cancer (EOCRC), defined as diagnosis before the age of 50 years, is increasing globally and is frequently characterized by aggressive biology and a disproportionate burden of liver metastases. This review synthesizes emerging evidence on the distinct molecular, immunologic and clinical features that differentiate EOCRC liver metastases from those arising in older adults. Genomic studies revealed increased chromosomal instability, increased copy number variation burden and unique amplification patterns involving MYC, RAD21, GNAS and MAPK1, alongside altered frequencies of classical driver mutations and increased germline predisposition. EOCRC liver metastases also exhibit a progenitor-like transcriptional state and an immune-cold microenvironment marked by reduced myeloid infiltration, impaired antigen presentation and profound resistance to immunotherapy, particularly in microsatellite-stable disease. Mechanistic insights into ferroptosis highlight therapeutic vulnerabilities, especially in PIK3CA-mutant tumors, where aspirin and ferroptosis inducers show synergistic potential. Clinically, high-risk EOCRC patients often present with left-sided primary tumors, synchronous metastases, adverse histology, elevated CEA levels and a hereditary predisposition, with prognostic models incorporating these variables outperforming traditional staging. Collectively, accumulating evidence suggests that EOCRC liver metastases may represent a biologically and clinically distinct entity, although ongoing debates regarding the extent of this distinction underscore the need for age-specific molecular profiling and prospectively validated therapeutic strategies.
Several studies have shown that excessive fructose consumption increases the risk of developing metabolic syndrome (MS), which can lead to diabetes mellitus. This study aimed to evaluate the metabolic and histological abnormalities accompanying MS induced by progressive high fructose diet (HFr) in Wistar rats. Twelve Wistar rats were used, divided into two groups (n = 6) (NC and HFr). They were fed a HFr with daily administration of increasingly higher doses of fructose solutions (dose 1 = 20% from week 1 to 4, dose 2 = 25% from week 5 to 8 and dose 3 = 30% from week 9 to 12 ), during which body weight and blood glucose were measured, while an oral glucose tolerance test (OGTT) was performed, it's glucose, triglyceride, and total cholesterol concentrations were determined, and pancreatic, liver, and kidney tissues were studied at the end of the experiment. Daily administration of increasingly high doses of fructose caused body weight increase, manifested by a highly significant difference in body weight, a metabolic abnormality, differences in fasting blood glucose (hyperglycaemia), plasma concentration of glucose, glucose intolerance, and by a very significant increase of total cholesterol and triglycerides levels in blood (hyperlipidaemia) between HFr and NC groups of rats. Histopathological study showed a slight swelling of the hepatocytes and a narrowing of the blood sinusoids, as well as an inflammatory infiltration of the portal space in the hepatic section of HFr group, no morphological alterations, necrosis, or steatosis in the pancreatic and renal sections of HFr group of rats. A twelve-week HFr diet can cause some abnormalities in the liver tissue, in carbohydrate and lipid metabolism in Wistar rats without damaging their pancreas and kidneys, which may be explained by the fact that metabolic abnormalities can precede histological lesions.
This study investigated the pathological and molecular effects of Centella asiatica extract-based cream and ointment formulations on burn wound healing in streptozotocin-induced diabetic rats. A total of 92 female Wistar albino rats were assigned to six groups: control (C), burn (B), diabetes (D), diabetes + burn (DB), diabetes + burn + cream (DBC), and diabetes + burn + ointment (DBP). Each group was further subdivided into 7-day (First subgroups) and 14-day (Second subgroups) treatment subgroups. Diabetes was induced by a single intraperitoneal injection of streptozotocin (55 mg/kg). Second-degree thermal burns (1 cm diameter) were induced on the dorsal skin of rats in the burn groups (B, DB, DBC, and DBP). Following burn induction, the DBC and DBP groups received daily topical treatment with a 2.5% Centella asiatica extract-based cream or ointment, respectively. Necropsies were performed on day 7 in the first subgroups and on day 14 in the second subgroups following burn model induction. Wound healing was evaluated macroscopically and histopathologically, and gene expression levels of TNF-α, FGF-2, EGF, VEGF, and IGF-1 were analyzed using quantitative real-time PCR. Wound areas were significantly larger in the DB group, whereas significant reductions were observed on days 7 and 14 in the treated groups (p < 0.05). Histopathological analysis revealed decreased inflammatory cell infiltration and enhanced fibroblast/fibrocyte density, angiogenesis, and epithelialization in the treated groups. Molecular analyses demonstrated significantly decreased TNF-α and increased FGF-2, EGF, VEGF, and IGF-1 expression in the treated groups compared to the DB group (p < 0.05). In conclusion, treatment with Centella asiatica extract supported healing in a diabetic burn model through anti-inflammatory, regenerative, angiogenic, and epithelializing effects. Notably, molecular analyses demonstrated a previously unreported increase in local IGF-1 levels following Centella asiatica extract treatment, supported by pathological findings.
BACKGROUND This report describes the case of an 81-year-old woman with a history of resected right-lung adenocarcinoma and presenting with a solitary nodule in the head of the pancreas, which demonstrated diagnostic challenges differentiating between metastasis of lung cancer versus primary pancreatic adenocarcinoma during the pathology workup. Differentiating between primary and metastasis in an isolated pancreatic lesion in a patient with a history of lung carcinoma is critical for patient treatment and management. Histology comparison, immunohistochemical characterization, and molecular studies, including next-generation sequencing (NGS) and microarray, are often utilized to differentiate between these two. CASE REPORT An 81-year-old woman with history of surgically resected right-lung adenocarcinoma in 2016 followed by adjuvant chemotherapy, presented in December 2024 with a solitary hypermetabolic pancreatic head nodule on positron emission tomography-computed tomography (PET CT). Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) of the pancreas lesion was consistent with moderately differentiated adenocarcinoma, showing overlapping histologic morphology and immunohistochemical profile with her prior lung adenocarcinoma. NGS identified genetic alterations favoring a lung origin. However, microarray analysis utilizing real-time RT-PCR done on the subsequently resected pancreas mass classified the lesion as more likely to be of gastrointestinal/pancreatobiliary, rather than lung origin. CONCLUSIONS Differentiating between primary versus metastasis from the lungs in a patient with an isolated pancreatic adenocarcinoma with a history of lung adenocarcinoma can be challenging, and occasionally a definite conclusion is still difficult to reach after comprehensive workups. Driver gene mutations, if present, which can be detected by NGS, may be more reliable in predicting tumor origin/type.
Osteoarthritis (OA) is a common degenerative joint disorder associated with aging, marked by chondrocyte senescence, cartilage degradation, and chronic inflammation. Although cellular senescence significantly contributes to OA progression, the molecular mechanisms governing this process are not fully understood. Interleukin-1 receptor-associated kinase 3 (IRAK3) acts as a negative regulator of Toll-like receptor (TLR) signaling; however, its role in OA and chondrocyte senescence remains unclear. We employed bioinformatics analysis, human OA cartilage samples, a destabilization of the medial meniscus (DMM) mouse model, and in vitro assays in ATDC5 chondrocytes to explore the expression and functional role of IRAK3 in OA. To evaluate senescence, mitochondrial function, reactive oxygen species (ROS), and inflammatory signaling, we conducted gain- and loss-of-function experiments alongside biochemical and histological analyses. IRAK3 expression was markedly reduced in both human and murine osteoarthritis (OA) cartilage. Silencing IRAK3 intensified hydrogen peroxide-induced senescence, mitochondrial dysfunction, reactive oxygen species (ROS) production, and catabolic metabolism, whereas its overexpression offered protective effects. Mechanistically, IRAK3 inhibited the TLR7/TLR9–NF-κB pathway, leading to decreased secretion of pro-inflammatory cytokines and reduced chondrocyte senescence. In vivo, intra-articular injection of adeno-associated virus (AAV) to knock down IRAK3 can promote the progression of OA in DMM mice. IRAK3 serves as a crucial inhibitor of chondrocyte senescence and the progression of osteoarthritis (OA) by reducing TLR-mediated inflammation and oxidative stress. These findings highlight IRAK3 as a promising therapeutic target for addressing senescence-related osteoarthritis.
Accelerated cellular senescence may be a key process in the progression of periodontitis, as it integrates the devastating effects of the major risk factors for periodontitis. Circadian rhythm disruption (CRD) affects the expression levels of multiple genes, such as brain and muscle ARNT-Like-1 (BMAL1), which is thought to be an important trigger or exacerbator of periodontitis. Even though CRD mechanisms are acknowledged to control cellular senescence, their effect on the senescence that happens during periodontitis is not well defined. This research aimed to explore the role and pathogenic mechanism of CRD in periodontitis and the involvement of cellular senescence, with the purpose of providing innovative ideas for the prevention and treatment of periodontitis. A rat model combining CRD and periodontitis was established. Periodontal lesions were assessed via histological staining. The expression levels of core circadian genes and senescence markers were evaluated. Inflammatory mediators related to the senescence-associated secretory phenotype (SASP) were quantified. The BMAL1 agonist SR8278 was employed to verify the key role of BMAL1 and the BMAL1/cryptochrome 2 (CRY2)/period circadian regulator 1 (PER1) signaling pathway. Finally, the effect of BMAL1 modulation on cellular senescence was examined in lipopolysaccharide (LPS)-induced human periodontal ligament cells (hPDLCs). CRD exacerbated experimental periodontitis lesions and aggravated the periodontal tissue senescence phenotype. BMAL1/CRY2/PER1 gene levels were down-regulated in a model of CRD-complexed periodontitis, and restoration of BMAL1 levels could alleviate CRD-exacerbated periodontitis by attenuating the periodontal tissue senescence phenotype. Interestingly, LPS exposure resulted in increased cellular senescence and decreased BMAL1/CRY2/PER1 in hPDLCs. Knockdown of BMAL1 resulted in further upregulation of cellular senescence in hPDLCs, whereas overexpression of BMAL1 inhibited LPS-induced cellular senescence. This study establishes a significant link between CRD and the aggravation of experimental periodontitis, within which a dysregulated BMAL1/CRY2/PER1 axis and an enhanced senescence phenotype are prominent features. This perspective opens new avenues for periodontitis intervention by focusing on circadian rhythm modulation.
Bellidifolin (BEL), the primary active compound in Gentianella acuta, demonstrates significant cardioprotective effects. This study aimed to investigate the protective role of BEL against DOX-induced myocardial injury and its mechanisms. Network pharmacology was utilized to identify possible therapeutic targets of BEL for disease treatment. In vivo, 30 C57BL/6 mice were randomly assigned to three experimental groups: Control, DOX, and BEL treatment (DOX + BEL). Mice were orally administered BEL (30 mg/kg) for 21 days, with a single intraperitoneal injection of DOX (15 mg/kg) on day 14. In vitro, H9c2 cells were used to assess the cardioprotective effects of BEL (50 µM) against DOX-induced toxicity. Cardiac function, myocardial histopathological changes, serum myocardial injury markers, oxidative stress indicators, and apoptosis were evaluated. Western blot analysis was conducted to detect the Nrf2/HO-1 and Galectin-3/NLRP3 pathway proteins. A total of 123 intersection targets were identified between BEL and disease-related proteins, including core targets such as caspase-3, IL-1β, and TNF. GO and KEGG pathway analyses revealed that BEL's protective effects against DOX-induced myocardial injury were associated with apoptosis, oxidative stress and inflammation. In vivo, BEL treatment improved cardiac function and myocardial histopathological morphology. BEL reduced MDA levels while increasing SOD and GSH-Px activity. Furthermore, BEL alleviated DOX-induced mitochondrial damage. BEL also decreased the apoptosis rate and modulated the expression of Caspase-3, Bax, and Bcl-2. Additionally, BEL activated the Nrf2/HO-1 pathway, as evidenced by increased nuclear translocation of Nrf2, upregulation of Nrf2, HO-1, GCLM, and NQO1 proteins, and a simultaneous decrease in Keap1 levels. BEL also suppressed the Galectin-3/NLRP3 inflammatory pathway, reducing the expression of Galectin-3, NLRP3, ASC, Caspase-1, IL-18, and IL-1β. In vitro, BEL treatment significantly decreased ROS levels induced by DOX. Overall, BEL mitigated DOX-induced cardiotoxicity by inhibiting oxidative stress, inflammation, and apoptosis via the Nrf2/HO-1 and Galectin-3/NLRP3 pathways.
Meningitis remains the leading infectious cause of neurological disabilities globally, disproportionately affecting children younger than 5 years and populations in the African meningitis belt. Whereas previous global estimates focused on ten pathogen categories, this study presents the most comprehensive analysis to date, assessing the meningitis burden attributable to 17 causative pathogens based on the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023 framework. GBD is a systematic, scientific effort aimed at quantifying the comparative magnitude of health loss caused by diseases, injuries, and risk factors across age groups, sexes, and geographical locations over time. We estimated meningitis mortality using the Cause of Death Ensemble model (CODEm) and morbidity using DisMod-MR 2.1, incorporating data from vital registration, verbal autopsy, surveillance, hospital data, and systematic reviews. Aetiology-specific estimates were generated with pathogen-linked case-fatality ratios and splined binomial regression models. Risk factor attribution was based on established risk-outcome pairs and population attributable fractions. In 2023, there were 259 000 (95% uncertainty interval 202 000-335 000) global deaths and 2·54 million (2·20-2·93) incident cases of meningitis. Children younger than 5 years accounted for more than a third of deaths (86 600 [53 300-149 000]). Streptococcus pneumoniae, Neisseria meningitidis, non-polio enteroviruses, and other viruses were the leading causes of death, while non-polio enteroviruses caused the most cases. The four WHO-defined preventable meningitis pathogens of interest (S pneumoniae, N meningitidis, Haemophilus influenzae, and Group B streptococcus) contributed to 98 700 deaths (77 000-127 000) and 594 000 cases (514 000-686 000). Low birthweight, short gestation, and household air pollution were the top risk factors for meningitis-related mortality. Although mortality and incidence have declined significantly since 1990, progress is insufficient to meet WHO 2030 targets. Despite marked progress in reducing bacterial meningitis via global vaccination campaigns, a substantial meningitis burden persists, attributable both to common pathogens such as S pneumoniae and N meningitidis and to emerging non-bacterial pathogens such as Candida spp and drug-resistant fungi. Achieving WHO goals will require sustained investment in surveillance, vaccination, maternal screening, and health-system strengthening, especially in high-burden settings. Gates Foundation, Wellcome Trust, and UK Department of Health and Social Care.
The relationship between actin dynamics and remyelination after intracerebral hemorrhage (ICH) remains unclear. This study aims to investigate the mechanism by which scalp acupuncture (SA) regulates remyelination in acute ICH rats through the PAK1/LIMK1/CFL1 signaling pathway. The Male Sprague-Dawley rats were used to establish the ICH model by autologous blood injection. The treatment group SA intervention, with the selective inhibitor of PAK1, IPA-3, used as a positive control. The regulatory effect of SA on the PAK1/LIMK1/CFL1 signaling pathway after ICH was evaluated using neurological function assessment, Luxol fast blue staining, Western blot, and immunofluorescence staining. On the third day after ICH, there is a peak in p-PAK1 levels, and SA intervention has the best therapeutic effect. Therefore, the third day was chosen for the IPA-3 positive control experiment. The study found that the PAK1/LIMK1/CFL1 signaling pathway is activated after ICH, leading to the inhibition of filamentous actin depolymerization, resulting in severe myelin sheath loss and neurological dysfunction. Following SA or IPA-3 intervention, the PAK1/LIMK1/CFL1 pathway is inhibited, leading to the depolymerization of actin filaments and a reduction in the degree of myelin sheath loss, as well as improvement in symptoms of neurological dysfunction. SA regulates the homeostasis of actin proteins by inhibiting the PAK1/LIMK1/CFL1 signaling pathway, promoting neuronal myelin sheath regeneration after acute ICH, and improving neurological behavioral disorders.
This study aims to evaluate the global burden of adverse effects of medical treatment (AEMT) using data from the Global Burden of Disease Study (GBD) 2021. Data were extracted from the GBD 2021, covering 204 countries/territories from 1990 to 2021. AEMT was defined using ICD-9 and ICD-10 codes, encompassing complications from medical procedures, treatments, or healthcare exposures. Estimates were categorized into fatal and non-fatal outcomes and stratified by age, sex, year, and covariates, including the Socio-demographic Index (SDI). Mortality-incidence ratios (MIRs), defined as the ratio of mortality calculated by dividing the number of deaths by the total incident cases, were analyzed. In 2021, the global age-standardized prevalence, incidence, disability-adjusted life years (DALYs), and mortality rates of AEMT were 11.48 (95% uncertainty interval [UI], 8.86-14.13), 150.44 (131.19-171.81), 64.19 (51.06-73.11), and 1.53 (1.29-1.68) per 100,000 population, respectively. DALY rates were highest in the early neonatal group (4,789.47 per 100,000 population [95% UI, 3,682.00-5,963.30]), while mortality rates followed a U-shaped pattern across age groups. In 2021, MIRs were highest at both ends of the age range: the early neonatal group (0.58 [95% UI, 0.55-0.58]) and the 95+ age group (0.05 [0.04-0.06]). This pattern was consistent across all SDI quintiles, with higher MIRs observed in lower SDI quintiles. The significantly higher prevalence and incidence rates of AEMT among the older population in high SDI quintiles, compared to lower SDI quintiles, could be attributed to the healthcare overutilization, highlighting the need for policy adjustments.
Bleomycin (BLM) is a chemotherapeutic agent that has been demonstrated to induce severe pulmonary and cardiac toxicity. Pirfenidone (PFD) is an established antifibrotic drug, while fisetin (FST) is a naturally occurring cardioprotective flavonol. The present study was conducted with the objective of evaluating the underlying mechanisms of BLM-induced cardiac damage and investigating the potential protective effects of combined PFD and FST treatment. Forty male Wistar albino rats were randomly divided into five groups (n = 8): Sham (intratracheal saline), and four groups that received a single intratracheal bleomycin injection (5 mg/kg) on day 1: BLM alone, BLM + PFD (50 mg/kg, oral), BLM + FST (25 mg/kg, oral), and BLM + PFD+FST (50 mg/kg PFD and 25 mg/kg FST, oral). Following induction, treatments were administered daily via gavage for 14 days. Cardiac damage was assessed using Hematoxylin and Eosin staining. ER stress-mediated apoptosis and oxidative stress were evaluated via immunohistochemistry (GADD153, Caspase-3), gene expression analysis (Bax/Bcl-2), and biochemical assays. The administration of BLM has been demonstrated to induce severe structural deterioration, characterised by the presence of persistent eosinophilic cardiomyocytes and haemorrhagic areas. While single treatments (BLM + PFD and BLM + FST) provided moderate improvements to histology, the combined treatment (BLM + PFD+FST) demonstrated superior preservation of tissue architecture. Biochemically and immunohistochemically, the BLM group exhibited a significantly increased Bax/Bcl-2 ratio, elevated caspase-3 and GADD153 expression, and exacerbated oxidative stress parameters. Conversely, the combined treatment significantly suppressed these pro-apoptotic signalling pathways and restored oxidative balance more effectively than either agent alone. Widespread cardiac fibrosis was not observed in any group during this acute phase. The combination of pirfenidone and fisetin in treatment provides superior cardiac protection against the toxicity induced by bleomycin in comparison to monotherapies. This is achieved by significantly attenuating pro-apoptotic signalling and restoring oxidative balance (GSH, MDA, MPO). These findings suggest that multi-targeted therapeutic approaches addressing both ER stress and oxidative pathways may be more effective than single-agent therapy in mitigating the cardiac complications associated with pulmonary fibrosis models.
Periodic fasting is known to improve metabolic health, but its impact on pancreatic islet plasticity remains unclear. We investigated the effects of intermittent fasting-mimicking diet (FMD) cycles on islet architecture and function in mice by performing immunohistochemical, ultrastructural, and metabolic analyses after fasting and after refeeding separately. Twelve-week-old female C57BL/6J mice were randomized to fasting (n = 9), refeeding (n = 10), or control group (n = 10). FMD was supplied weekly for 3 days (50%, 10%, 10% of daily caloric intake) followed by 4 days of chow food ad libitum (except for pre-IGTT food withdrawal and the duration of the IGTT). Intraperitoneal glucose tolerance tests (IGTTs) were performed at day 11 (fasting group), day 14 (refeeding group), and day 13 (control group). Mice were sacrificed 7 days after IGTT, and pancreata were subjected to fluorescence immunohistochemistry or scanning electron microscopy (STEM). Bodyweight, blood glucose, proinsulin, and IGF-1 concentrations were significantly decreased after fasting but rebounded after refeeding. Pancreatic insulin+glucagon+, BRN4+, and PDX1+BRN4+ cells increased significantly after fasting and tended to remain high after refeeding, thereby indicating increased pancreatic islet plasticity after fasting. In STEM images, the insulin granule core-to-halo ratio increased significantly after fasting. The fasting but not the refeeding group showed impaired glucose tolerance. The more crystallized mature β-cell granules indicate increased insulin secretory capacity, and the reduced proinsulin-to-insulin ratio suggests reduced endoplasmic reticulum stress in ß-cells after fasting. We propose that this observed plasticity may provide a basis for novel concepts of in vivo β-cell regeneration. However, further studies to investigate molecular mechanisms of fasting/refeeding in murine type 1 diabetes to evaluate its therapeutic potential are needed.
This article, the second in a series of eight, highlights the lives and original works of 21 scientists whose names are preserved in 20 enduring eponyms still found in dental anatomy and histology textbooks. Though frequently referenced in education, the historical context and the original publications behind these terms are often overlooked. By revisiting their biographies and citing the original sources where each eponym was first described, this work offers a long-overdue acknowledgment of their lasting contributions to dental science.
Emerging evidence suggests a crucial role of MARCH8, a membrane-associated RING-E3 ubiquitin ligase, in cancer progression by regulating the turnover of cancer-related proteins. Our previous research identified potential MARCH8 targets using an RNAi-coupled proteomics approach, revealing that MARCH8 mediates the proteasomal degradation of E-cadherin and β2m, contributing to Esophageal Squamous Cell Carcinoma (ESCC) progression. Additionally, EpCAM emerged as a key target, and this study aims to investigate how MARCH8 regulates EpCAM stability and its implications in tumor migration. Herein, Co-immunoprecipitation (Co-IP) and Western Blotting (WB) demonstrated that MARCH8 directly interacts with EpCAM and regulates its turnover by ubiquitination and proteasomal degradation. Further, MARCH8-EpCAM co-localization was observed in ESCC cells using Immunofluorescence (IF). Additionally, Immunohistochemistry (IHC) analysis in ESCC tissues revealed a significant inverse correlation between MARCH8 and EpCAM expression (r = -0.6730, p < 0.0001). Interestingly, EpCAM showed a context-dependent expression in esophageal tissues. Adjacent non-malignant esophageal squamous epithelium showed no detectable EpCAM staining, superficial tumor regions displayed membranous EpCAM expression, whereas tumor cells at the deeper invasive front exhibited markedly reduced or lost EpCAM expression. Consistently, scratch assay coupled with IF and IHC analysis further demonstrated reduced EpCAM expression in migrating ESCC cells. Overexpression studies further revealed an inverse relationship between MARCH8 and EpCAM in migration, invasion, and β-catenin signaling, as shown by invasion/migration assays and WB analysis. Collectively, these findings establish EpCAM as a novel target of MARCH8. Given that loss of EpCAM is associated with migratory phenotypes and that MARCH8 promotes ESCC progression, MARCH8-driven EpCAM degradation may contribute to enhanced tumor cell migration and disease progression.
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A histological safety profile is essential for non-antibiotic microbial interventions in aquaculture. This study assessed the multi-organ histological biocompatibility of a novel actinobacterial strain, Agrococcus sp. RKDAS_1, in Nile tilapia (Oreochromis niloticus) under controlled conditions. Juvenile tilapia were fed diets with varying concentrations of RKDAS_1 (105, 107, and 109 CFU g- 1 feed) for 60 days. Tissues from the gill, intestine, liver, and heart were analysed through standard histological methods, employing semiquantitative scoring and intestinal morphometry. Across all examined organs, RKDAS_1 supplementation did not induce inflammatory responses, degenerative lesions, or structural disruptions, which are indicative of tissue-level toxicity. Gill architecture was intact, with normal hepatocyte arrangements and no necrosis or fibrosis, while cardiac tissues showed a normal structure. Intestinal morphology maintained epithelial integrity, displaying dose-related variations in villus height and goblet cell density. Intermediate-dose live hepatic sections showed reduced cytoplasmic vacuolation compared to controls, though the difference was not statistically significant. The semi-quantitative histopathological evaluation showed that the tissue structure remained intact across different treatments. The results collectively suggest that Agrococcus sp. RKDAS_1 did not cause any noticeable histopathological damage, indicating enhanced biocompatibility. The findings suggest that Agrococcus sp. RKDAS_1 is compatible with biological tissues without causing significant damage. However, these conclusions are restricted to structural analysis and do not ensure functional improvements or overall safety beyond tissue examination. The findings provide a critical starting point for future research, which will delve into molecular, immunological, and long-term exposure studies. These investigations aim to evaluate the biotherapeutic potential of RKDAS_1 thoroughly.
Polycystic ovary syndrome (PCOS), one of the leading causes of anovulatory infertility, is a complex endocrinopathy influenced by environmental factors. Evidence suggests a connection between PCOS and mammalian target of rapamycin (mTOR), a serine/threonine kinase involved in cellular growth, proliferation, and metabolism. MicroRNA-21(miR-21), a key post-transcriptional regulator of ovarian function, is also implicated in follicular development, atresia, and steroidogenesis. This study aimed to investigate the relationship between miR-21 and mTOR in an experimental PCOS mouse model.Forty-two Balb/c female mice (25-day-old) were divided into six groups: Control, SO (PCOS vehicle-control), PCOS, Inhibition, PCOS+Inhibition, and DMSO (mTOR inhibition-vehicle-control. PCOS was induced via subcutaneous injection of dehydroepiandrosterone (6 mg/100 g), and mTOR inhibition was achieved with KU-0063794 (1 mg/100 g). Serum estrogen (E2) and progesterone (P4) levels were measured by ELISA. Ovarian tissues were evaluated histomorphologically and immunohistochemically.Compared with controls, PCOS mice exhibited increased body weight (p < 0.05), elevated serum E2 (p < 0.001), P4 (p < 0.01) and cystic follicles, reduced corpora lutea (p < 0.001), and enhanced mTOR/p-mTOR and PCNA immunoreactivity (p < 0.001). In the PCOS + INH group, mTOR and p-mTOR immunoreactivity were significantly reduced compared with PCOS (p < 0.001), and DHEA-induced weight gain was attenuated; however, E2 remained elevated (p < 0.001) and corpora lutea were not restored (p < 0.001). miR-21 expression was significantly upregulated in both PCOS (p < 0.05)-PCOS + INH (p < 0.001) groups, indicating persistent miR-21 activation despite mTOR inhibition.These findings demonstrate that while mTOR signaling is activated in PCOS and may be pharmacologically suppressed, mTOR inhibition alone is insufficient to normalize steroidogenesis, ovulation, or miR-21 expression, suggesting mTOR-independent regulation of miR-21 in this model.