Ebola virus (EBOV) infection typically results in severe-and often lethal-acute disease. However, increasing evidence suggests that EBOV can persist in certain immune-privileged tissues, which may then serve as reservoirs for the later reemergence of EBOV and disease recrudescence. Here, we report atypical EVD recrudescence in a ferret model inoculated with an otherwise lethal dose of EBOV and treated with low doses of a highly potent monoclonal antibody cocktail. Among 32 antibody-treated ferrets, 14 animals survived, while 8 succumbed to acute EVD within about 5-8 days. The remaining 10 animals succumbed to atypical EVD between 12 and 18 days post-infection (DPI) despite having shown no, or very minor, signs of illness during the acute phase of disease. While viremia disappeared by 14 DPI in most animals that succumbed to atypical EVD, it rebounded modestly just prior to death. Unlike animals that died of acute EVD, those that died of atypical EVD showed only a moderate systemic inflammatory response and few signs of organ dysfunction, in line with low levels of virus in the liver and spleen. Interestingly, however, ferrets that died of atypical EVD showed high levels of virus in the brain, consistent with increased markers of inflammation in the central nervous system and significant pathological changes, including a breakdown in the blood-brain barrier and severe meningoencephalitis. Not only does this study shed important light on the atypical and underappreciated manifestations of EVD, but it also establishes the ferret as a valuable model of EBOV recrudescence.
Hookworm infections continue to impose a substantial burden on human and animal health, but the early host responses that influence parasite establishment are not fully characterized. Experimental models that reproduce key features of hookworm biology and host-parasite interactions remain essential for advancing translational research. In this study, we examined hematological, biochemical, immunological, and parasitological parameters during the acute phase of experimental hookworm infection using the Ancylostoma ceylanicum-Mesocricetus auratus model, a small-animal system widely employed for mechanistic studies of hookworm infection. Animals were evaluated at 7 and 20 days post-infection. Hematological indices and serum iron concentrations did not differ between infected and control groups during the acute phase. In contrast, infected animals showed increased splenic mass at 20 days post-infection, indicating immunological activation. Hepatic hepcidin expression was markedly reduced, suggesting an early alteration in systemic iron regulation. Analysis of inflammatory mediators revealed selective modulation of cytokine expression, with reduced interleukin-6 transcript levels at 20 days post-infection, whereas tumor necrosis factor alpha expression remained unchanged. Parasitological analyses demonstrated progressive parasite establishment, with fecal egg output detected from 14 days post-infection and reaching approximately 300 eggs per gram by day 18, consistent with the onset of patency. Taken together, these data indicate that acute hookworm infection induces coordinated changes in immune responses and iron metabolism before the development of overt hematological alterations.
Crimean-Congo haemorrhagic fever (CCHF) is a severe, widespread, tick-borne viral zoonotic infection. It is caused by an orthonairovirus that is transmitted by ticks. Sero-epidemiological studies in humans and livestock are valuable indicators of viral circulation and infection risk. This study aimed to investigate the seroprevalence and factors associated with CCHF virus exposure in humans and livestock in mixed crop-livestock farming households in rural Burkina Faso. A cross-sectional animal-human linked study was conducted in 149 rural households across 16 randomly selected villages in two administrative regions of Burkina Faso. Human socio-demographic, livestock biodata, and serum samples were collected from household members and their livestock (cattle, sheep, and goats). Additional ecological and climatic data were extracted from online databases and merged with the field data. Serological testing was performed on human and animal samples using the ID Screen® CCHF Double Antigen Multi-species ELISA (IDvet, Grabels, France). Descriptive statistics and multivariable multilevel analyses were used to assess factors associated with exposure of cattle and small ruminants to CCHF virus, while the Fisher's exact test was applied to assess the risk factors for human exposure. The study included 717 livestock farmers and 2,295 animals, comprising 666 cattle, 659 sheep and 970 goats. The overall CCHF virus (CCHFV) seroprevalence was 3.1% (95% CI: 1.9-4.6) in humans and 54% (95% CI: 50.2-57.7) in cattle. In small ruminants, the overall seroprevalence was 5.2% (95% CI: 4.2-6.4), with 9.1% (95% CI: 7.1-11.5) in sheep, and 2.5% (95% CI: 1.7-3.8) in goats. Farmers with inadequate livestock management-related biosecurity behaviour exhibited higher seroprevalence rates and an increased risk of CCHFV seropositivity. In cattle, seropositivity was positively associated with older age, female sex, longer grazing distances, and tick infestation. Seropositivity in small ruminants was associated with older age, being of the sheep species, and longer grazing distances. Ecological factors, including a higher aridity index in both cattle and small ruminants, and steeper slopes in cattle, were significant in univariate and multivariable analysis, respectively. The seroprevalence in both cattle and small ruminants showed significant clustering within households, with intra-cluster correlation (ICC) rates of 39% and 62%, respectively. This study highlighted that CCHFV is circulating among humans and their livestock in rural Burkina Faso. Individual and household-related risk factors, including socio-demographic, livestock management practices, and ecological characteristics, were identified. These findings provide valuable insights for designing tailored public health interventions towards strengthening CCHF surveillance and prevention among rural households.
The global rise in pet ownership has increased demand for health-promoting products, particularly probiotics designed to support gastrointestinal and immune health in companion animals. However, most commercial products rely on non-host-adapted strains, which may limit gastrointestinal colonization and host-specific benefits. To address this gap, 56 bacterial isolates were obtained from the fecal and milk microbiota of clinically healthy dogs and cats. Among these, Limosilactobacillus reuteri DF/KS2, derived from the fecal microbiota of a Kangal Shepherd dog, and Enterococcus faecium CM/BS2 derived from the milk microbiota British Shorthair cat, were selected based on their broad-spectrum antimicrobial activity. Both strains exhibited a safe profile, as evidenced by γ-hemolysis and susceptibility to a panel of clinically relevant antibiotics. Under simulated gastrointestinal conditions, CM/BS2 and DF/KS2 tolerated highly acidic environments and demonstrated resilience against digestive enzymes and bile salts. Furthermore, both isolates displayed strong auto-aggregation and co-aggregation abilities with key pathogens, while adhesion assays using Caco-2 cells confirmed their capacity to inhibit pathogen attachment. Immunomodulatory evaluations further revealed that both strains effectively reduced pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) and enhanced anti-inflammatory IL-10 production in canine and feline macrophages. Optimal growth occurred at 37 °C after 24 h in 2% molasses medium, and shelf-life studies demonstrated that freeze-dried cultures retained high viability over six months at - 20 °C. Collectively, these findings highlight the probiotic potential of host-adapted L. reuteri DF/KS2 and E. faecium CM/BS2, emphasizing their suitability for inclusion in species-specific probiotic formulations aimed at supporting gastrointestinal and immune health in dogs and cats.
Crimean-Congo hemorrhagic fever (CCHF) is an emerging tick-borne disease with a growing global distribution. In Armenia, 31 Ixodidae tick species have been identified, including Hyalomma species. While CCHF virus (CCHFV) was previously isolated from ticks in the 1970s, no further studies were conducted until 2016, when antibody to CCHFV was identified.This study aimed to update data on the Ixodidae tick species, detect CCHFV in collected specimens, and perform molecular characterization of viral lineages circulating in Armenia during 2022-2024.Epizootiological investigations were performed across all 10 regions of Armenia. Fieldwork involved the collection of Ixodidae ticks, followed by morphological identification and testing of specimens. A total of 550 animals (including cattle, sheep, and horses) and surrounding open habitats were surveyed, resulting in the collection of 3158 Ixodidae ticks representing 12 species. These samples were organized into 960 pools and tested via reverse transcriptase polymerase chain reaction for CCHFV. Viral RNA was detected in 96 pools, all originating from ticks collected from cattle and representing 8 species. Positive samples were confined to two regions: Syunik and Tavush. Phylogenetic analysis revealed the presence of two CCHFV lineages in Armenia: Europe 1 (Subgroup V) and Europe 3 (Subgroup VII).Armenia's climatic and geographic diversity, expanding land use, population mobility, and growing tourism highlight the need for continued surveillance. The present identification of CCHFV in ticks in two regions emphasizes the importance of ongoing epizootiological monitoring and molecular characterization to assess the broader epidemic risk across the country.
Cellulose is the most abundant natural biopolymer obtained from plants, animals, and microbes. Cellulose is porous, biocompatible, biodegradable, noncytotoxic, and can retain water for a longer period. Therefore, cellulose has various applications in the medical, cosmetic, paper, and food industries. The current review summarizes the recent progress related to the application of cellulose in skin biology. Cellulose is preferable for skin biology applications due to its porosity, and hygroscopic nature that enables it to mimic the extracellular matrix (ECM) of native skin. Cellulose can be modified by combining with other biopolymers to make a functional cellulose composite. Researchers have developed various cellulose composites that are successfully used as wound dressing agents, scaffolds for skin tissue engineering, and vehicles for drug delivery like anticancer or other hydrophobic drugs. Cellulose-based facial masks act as a carrier for active agents like anti-aging, anti-wrinkle, and anti-acne substances. Cellulose can also be used as a stabilizer in the preparation of emulsion-based creams and other cosmetic products. It is concluded that cellulose is a suitable biopolymer in the global marketplace for skin applications, but further advancements should be made to maximize its clinical use.
Aging, trauma, genetic predisposition, and lifestyle impact the human body´s cartilage degradation. Once injured, continuous insults resulting from daily activities can trigger pathological conditions such as osteoarthritis, the primary cause of disability and socioeconomic loss worldwide. The main goal of all orthopedic surgeons treating joint cartilage injuries has been to reduce the extent of the lesion anatomically, repair the cartilage surface, and reestablish joint stability. Animal models are essential for the development of therapeutic drugs, but current models for cartilage defects are unsatisfactory. Osteochondral lesions in sheep are a valuable model for testing new therapies and biomaterials that can aid in the recovery of cartilage and bone in human joint environments. This study established an efficient protocol for inducing acute osteochondral defects in large animals. A standardized lesion was created in both medial femoral condyles. One knee was randomly assigned to receive gelatin-methacryloyl treatment, while the contralateral knee served as a control. Six months after the surgical procedure, the femoral condyle area was removed, the dissected knee joints were decalcified, embedded in paraffin, and cut into sections, which were stained with hematoxylin and eosin. Scores were used to evaluate the lesion. This methodology allows immediate macroscopic observation after injury induction. Additionally, this model effectively replicates clinical cartilage defects, providing a valuable model for studying their pathology and developing innovative therapeutic approaches.
Giardia duodenalis is a major intestinal protozoan parasite responsible for over 200 million infections annually worldwide. The gut microbiota influences parasite colonization, disease outcome, and host immune responses. In this context, the present study aimed to evaluate Th17-associated cytokine levels during Giardia duodenalis infection. This study evaluated the influence of gut microbiota modulation during Giardia duodenalis infection by analyzing Th17-related cytokines (IL-17 A, IL-17 F, IL-22, IL-23, and IL-10) in antibiotic pre-treated, Lactobacillus pre-treated, and untreated Giardia-infected mice. Serum cytokine profiles were also assessed in Giardia-infected patients to examine their association with clinical severity. Mice were divided into antibiotic pre-treated, probiotic (Lactobacillus) pre-treated, and untreated groups prior to Giardia infection. Animals were sacrificed on day 0 and on days 3, 7, and 14 post-infection. Th17 cytokines were quantified by ELISA, parasite burden was determined by RT-PCR, and intestinal pathology was assessed by hematoxylin and eosin staining. Additionally, 60 Giardia positive patients were clinically evaluated using the Vesikari scoring system and classified as mild and moderate to severe symptoms. Serum Th17 cytokines were measured by ELISA. Giardiasis increased both pro and anti-inflammatory cytokines in untreated mice and antibiotic pre-treatment exacerbated giardiasis, resulting in increased parasite load, heightened inflammatory cytokine responses, and severe intestinal mucosal damage. In contrast, probiotic pre-treatment induced an early and regulated cytokine response, reduced parasite burden, promoted mucosal healing, and facilitated faster recovery. Clinically, patients with moderate to severe disease exhibited elevated IL-17 A, IL-17 F, IL-23, and IL-10 levels, whereas higher IL-22 levels were associated with mild symptoms, suggesting a protective role. Antibiotic-induced dysbiosis aggravates giardiasis, while probiotic modulation of gut microbiota enhances protective immunity and reduces disease severity, highlighting probiotics as potential preventive and therapeutic agents.
Inflammatory bowel disease (IBD) is driven by a breakdown in immune regulation and epithelial barrier function, yet the contribution of eosinophils to this process has remained poorly defined and controversial. While eosinophils infiltrate the intestinal mucosa during both flares and remission, their role in shaping disease outcomes is unclear. Our RNA-seq analyses of colonic eosinophils isolated from dextran sulfate sodium (DSS)-treated mice revealed a significant upregulation of cyclooxygenase (COX)-2 (gene name, Ptgs2). Eosinophil-specific deletion of COX-2 (Ptgs2fl/fleoCre+/-) reduced IL-22 production and exacerbated DSS- and trinitrobenzene sulfonic acid (TNBS)-induced colitis, characterized by greater weight loss, higher disease activity, colon shortening, and epithelial injury. Administration of recombinant IL-22 reversed these phenotypes. Mechanistically, eosinophil-derived COX-2 enhanced IL-22 production by type 3 Innate lymphoid cells (ILC3s) through prostaglandin E2 (PGE2) signaling. Consistently, Ptgs2fl/fleoCre+/- mice exhibited reduced colonic PGE2 levels, while PGE2 analog treatment restored IL-22 production and mucosal protection. Our findings identify eosinophil-derived COX-2 and PGE2 as a critical regulator of IL-22 production during colitis, uncovering a eosinophil-ILC3 cross talk that safeguards the intestinal barrier and represents a promising therapeutic target in IBD.
Colorectal cancer (CRC) and cardiovascular disease (CVD) are major contributors to the global disease burden, with CRC as the third most common malignancy and second leading cause of cancer-related death, and CVD as the top killer among non-communicable diseases. The growing ageing population has driven a continuous rise in CRC-CVD comorbidity. Treatment-related cardiovascular toxicity, shared pathophysiological links, and the prognostic impact of comorbidity have made this interdisciplinary field a critical focus in cardio-oncology. This review addresses core scientific questions at the CRC-CVD intersection, including their shared risk factors and pathological mechanisms (metabolic syndrome, chronic inflammation, oxidative stress, gut microbiota dysbiosis, etc.), epidemiological and molecular associations between CRC and major CVD subtypes, as well as cardiovascular toxicity mechanisms, monitoring, and intervention strategies for CRC therapies (chemotherapy, targeted therapy, immunotherapy). Beneficial lifestyle modifications and cross-effective medications, together with multidisciplinary team (MDT) collaboration, hold great value for comorbidity management. Current studies have confirmed their epidemiological and partial mechanistic links, but key gaps remain: unclear core molecular regulatory networks, insufficient gut microbiota-cardiovascular-tumor axis research, lack of personalized risk prediction models, limited long-term cardiovascular data for novel anticancer drugs, and absent standardized precision prevention protocols. Future multi-omics studies, large-scale clinical trials, and real-world data research will help clarify shared targets, build risk models, and formulate interdisciplinary management guidelines to improve outcomes of CRC-CVD comorbidity patients.
The detrimental cycle of sarcopenic obesity (SO) significantly reduces quality of life in older adults, while the mechanisms are still unclear. We first analyzed the incidence of SO using the CHARLS database. We identified key genes by integrating differentially expressed genes, weighted gene co-expression network analysis, and targets of gut microbiota metabolites, refining the selection through machine learning methods (LASSO, XGBoost, SVM-REF, Random Forest). These genes were validated through single-cell sequencing, receiver operating characteristic analysis, and Muscle immunohistochemistry in a high-fat-diet (HFD) induced mouse model. Further analyses comprised immune infiltration profiling, pathway enrichment, and transcriptional regulation analysis. Additionally, we explored the relationships between key genes and autophagy, ferroptosis, and immunity responses. Finally, we predicted and evaluated potential therapeutic compounds via the CMap database and molecular docking. SO incidence in China increased significantly from 16.1% (2011) to 20.4% (2018). Machine learning identified ALDH1A3, CSF1R, and PHGDH as key genes. These genes were validated in external muscle single-cell datasets, demonstrating robust diagnostic performance with AUC values exceeding 0.72 across four independent GEO cohorts. Following an HFD intervention in mice, ALDH1A3 and CSF1R expression in muscle tissue was significantly upregulated, while PHGDH showed a consistent upward trend that did not reach statistical significance. Immune infiltration analysis revealed a significant increase in resting NK cells in both obesity and sarcopenia states. Functional enrichment analyses using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes linked the genes to transcriptional regulation pathways. The Cisbp_M4923 motif was identified as the most relevant transcription factor binding site. Finally, molecular docking simulations indicated stable binding of the top candidate compound, Birinapant, to the key gene targets. ALDH1A3, CSF1R, and PHGDH serve as potential co-morbid biomarkers for SO.
Laser capture microdissection (LCM) provides spatial access to specific cell populations within complex tissues through in situ visualization and isolation. To enable transcriptomic analysis of histologically defined regions in fixed tissue, a detailed LCM protocol is presented for RNA extraction from paraformaldehyde (PFA)-fixed, Optimal Cutting Temperature (OCT) compound-embedded mouse liver sections. The protocol details the identification and collection of microscale samples (approximately 1,000 cells) through a workflow encompassing tissue fixation, sucrose dehydration, OCT embedding, cryosectioning, hematoxylin staining, and laser-capture of targeted histological areas. Using this method, a high-purity RNA (A260/A280: 1.9-2.1) was obtained. The RNA integrity number (RIN) was 6.7 ± 0.9, reflecting the expected fragmentation associated with PFA fixation. However, quantitative PCR for β-actin yielded Ct values of 17-19, and RNA sequencing performed using fragmentation-optimized library preparation generated high-quality reads, with >90% of bases meeting Q20 and Q30 thresholds, confirming that the RNA is suitable for sensitive downstream analyses. Therefore, this protocol enables spatially resolved, targeted gene expression analysis by providing RNA of defined purity and integrity from specific histological regions of PFA-fixed liver tissue.
Effective pest management requires accurate and continuous monitoring. This monitoring helps assess population dynamics and guides the development of integrated pest management strategies. Traps used to capture insects are an alternative applied to various crops. However, the identification and manual counting of specimens are time-consuming, require taxonomic knowledge, and depend on the expertise of specialists. Automation could reduce costs, increase accuracy, and enable scalable analyses. Current computer vision and artificial intelligence techniques can quickly and accurately identify objects in digital images. This study presents a systematic review of literature retrieved from multidisciplinary and specialized databases (Scopus, ACM, Web of Science, IET, DBLP, Springer, and ScienceDirect), focusing on the intersections of agriculture, ecology, and computer science. We found 284 studies published between 2020 and 2025. Among them, 57 fulfilled the eligibility criteria, considering applied computing solutions for insect identification and counting using digital images of specimens collected via traps or photographed in situ on plants, in both field and laboratory settings. The findings highlight the use of electronic traps for real-time data collection and improvements in convolutional neural networks, with visual transformers and attention mechanisms for multi-species and fine-grained recognition. They also indicate opportunities to leverage microscopy resources, overcome limitations in the large-scale deployment and integration of electronic trap networks, and integrate real-time monitoring data with forecasting models using weather predictions to promote early warning systems for integrated pest management.
Chromatin accessibility plays essential roles in transcription, DNA repair, and chromosome segregation. Hyper-accessible regions usually correlate with active promoters and enhancers, facilitating transcription factor binding and regulatory activity. The assay for transposase-accessible chromatin using sequencing (ATAC-seq) enables genome-wide profiling of chromatin accessibility with very few cells. However, its implementation in Caenorhabditis elegans is limited by the nematode's rich collagen cuticle that complicates cell dissociation. Here, we present an optimized protocol for performing ATAC-seq in whole worms at the L4 stage. The procedure begins with synchronized cultures and involves cuticle disruption, enzymatic dissociation, and cell-suspension preparation. Permeabilized nuclei are then subjected to Tn5 transposition, followed by polymerase chain reaction (PCR) amplification and purification of next-generation sequencing (NGS)-ready libraries. This protocol requires 30 µL of worm pellet, can be completed in one day, and generates 5,000-9,000 accessibility peaks in the Bristol N2 reference strain. This streamlined workflow can be adapted to other developmental stages or FACS-purified cell populations. By reducing technical barriers to ATAC-seq in C. elegans, this method expands opportunities to study genome-wide chromatin accessibility in response to genetic and environmental perturbations in a whole-organism context.
The microbiome is a key determinant of organismal health, yet inter-individual variability and heterogeneous responses to environmental conditions complicates the understanding of its effects on hosts. Here, we present a comprehensive analysis using the Drosophila Genetic Reference Panel (DGRP) to investigate how the interplay between host genetic variation and diet influences microbiome composition, and to assess whether microbiome features in young flies can be used to predict lifespan and age-related traits. Our findings show that adult flies reared on a nutritionally rich control diet exhibited higher microbial richness but lower evenness compared to those on a nutritionally poor restricted diet. Principal component analysis (PCA) highlighted substantial diversity among lines reared on the same diet, and this variation was evidenced by high heritability estimates for all measured α-diversity metrics, including Unique OTU counts, Shannon and Simpson indices, as well as the relative abundances of genera and species with relative abundances exceeding 1%. These results underscore the critical roles of both environmental factors and genetic variation in shaping microbiome composition under different dietary conditions. Moreover, we identified widespread genotype-by-diet interactions, suggesting that the genetic regulation of the microbiome is highly complex. Finally, we found that the microbiome features of young flies including diversity indices, taxonomic abundances, or ordination scores cannot predict age-associated phenotypes (lifespan, locomotor activity, dry weight, and heat knockdown time). Our findings offer valuable insights into the genetic architecture that governs microbiome composition, dietary responses, and aging in Drosophila melanogaster.
Germ cells of diverse species form a syncytium, which is a multinucleated cell complex, and share cytoplasmic components via intercellular bridges. Here, we demonstrate that the paralogous transmembrane proteins ROOM-1 and ROOM-2, identified through the proteomic analysis of maternal membrane proteins, are essential for the maintenance of germ cell compartments from the rachis of Caenorhabditis elegans gonads. Although animals lacking each protein are fertile, the loss of both proteins resulted in complete sterility due to compartmentalization failure of individual germ cells in the late larval and adult gonads. Additionally, the ROOM proteins colocalize specifically to the intercellular bridge with F-actin and ANI-2, a protein organizing the intercellular bridge of germ cells. The localizations of ANI-2 and ROOM-1/2 on the rachis bridge were interdependent. Thus, the transmembrane ROOM proteins redundantly function to ensure "rooms" for germ cells by maintaining the intercellular bridges.
Brain metastasis remains a devastating clinical problem. A major challenge in brain metastasis research is the lack of high-quality models that accurately recapitulate the metastatic process, thereby enabling mechanistic insights into how cancer cells colonize in the brain. Traditional intracarotid artery injection models of brain metastasis often require permanent ligation of the common carotid artery (CCA), which alters cerebral hemodynamics and compromises the integrity of the blood-brain barrier (BBB). The protocol presents a refined method for establishing a high-fidelity mouse model of brain metastasis. The core innovation involves the Interlock Pulsatile Injection (IPI) technique for tumor cell delivery, followed by microsurgical arterial reconstruction at the puncture site to restore physiological blood flow in the CCA. Compared with the conventional CCA ligation model, the IPI-microsurgical repair approach significantly reduced perioperative mortality (2.86% vs. 25.71%) and increased the rate of brain metastasis establishment (94.12% vs. 65.38%). The IPI technique utilizes a tandem syringe configuration to minimize cell regurgitation during intracarotid injection. After tumor cell infusion, the CCA is meticulously repaired using microsurgical sutures under a stereomicroscope, thereby avoiding permanent occlusion. This preserves the native cerebral hemodynamics and the integrity of the BBB at the time of tumor cell entry, while significantly improving surgical success rates and reducing mortality. The metastatic intracranial lesions are validated by serial bioluminescence imaging and histopathology. The method provides a superior platform for studying the pathophysiology of brain metastasis and for preclinical therapeutic evaluation, thereby recapitulating the metastatic process.
Data on feline hemotropic mycoplasmas is limited in Iran. The aim of this study was to investigate the infection rate of cats with this pathogen in Mashhad,Iran and evaluate its assosiation with cliniopathological changes. Blood samples from 100 cats presenting at veterinary clinics were analysed for Mycoplasma haemofelis (Mhf) and Candidatus Mycoplasma haemominutum (CMhm). Samples were screened for hemotropic mycoplasmas using microscopic examination and conventional polymerase chain reaction (PCR). Samples were also subjected to haematological and biochemical analysis. Microscopic examination detected hemotropic mycoplasmas in 65% of samples, whereas PCR identified a 23% infection rate, with 65.21% (n = 15) positive for CMhm and 34.78% (n = 8) for Mhf. The presence of Haemomycoplasma DNA was significantly associated with age, gender, breed and roaming status (p < 0.05). Infected cats exhibited significant alterations (p < 0.05) in haematocrit (Hct), red blood cell count (RBC), haemoglobin (HGB), white blood cell count (WBC), neutrophil counts, serum total protein (TP), globulin, phosphorus (Pi), aspartate aminotransferase (AST) levels and the albumin/globulin ratio (AGR). Our study highlights a high prevalence of feline hemoplasmosis in northeastern Iran, with CMhm as the predominant species. Key risk factors included male gender, adult, outdoor access and domestic short hair (DSH) breed. Although infected cats showed consistent clinicopathologic changes, no differences were observed between Mhf and CMhm infections.
Metformin, widely used for type 2 diabetes mellitus, has demonstrated antitumor effects through modulation of metabolic and immune pathways. This review explores its potential role in the prevention and treatment of skin cancers, including melanoma and nonmelanoma skin cancers (NMSCs). A structured PubMed search was conducted in April 2025 to identify English-language, peer-reviewed original research articles evaluating the effects of metformin on melanoma, basal cell carcinoma (BCC), and squamous cell carcinoma (SCC). Experimental, observational, and clinical studies were included; reviews and meta-analyses were excluded. Twenty studies met the inclusion criteria. In melanoma models, metformin inhibited tumor proliferation, suppressed epithelial-mesenchymal transition, and enhanced immune responses. Observational studies reported improved recurrence-free survival and treatment outcomes, although findings were inconsistent. In NMSC studies, metformin use was associated with reduced incidence of BCC and SCC. Preclinical models demonstrated delayed tumor development and an enhanced response to photodynamic therapy following metformin treatment. Mechanistic and preclinical data support a biologically plausible role for metformin in skin cancer prevention and therapy. Evidence is strongest for BCC, particularly in enhancing photodynamic therapy and reducing incidence in at-risk populations. Melanoma studies suggest synergy with immunotherapy, but clinical results remain variable. Metformin shows promise as a low-cost, well-tolerated adjunctive therapy in dermatologic oncology. Further prospective and controlled studies are needed to clarify its efficacy, optimize dosing, and identify populations most likely to benefit.
The purpose of this study is to identify hub genes associated with both osteoporosis (OP) and chronic kidney disease (CKD) through bioinformatics analysis, and to explore the potential pathogenetic mechanisms in OP and CKD through these hub genes. We downloaded the GSE15072 and GSE56815 datasets from the GEO database as training sets, and GSE7158 and GSE70528 for validation. Differential expression genes were selected using the "limma" package, while gene co-expression networks were constructed with "WGCNA." Functional enrichment analyses were performed using "clusterProfiler." Hub genes were identified through machine learning techniques, and their diagnostic efficacy was evaluated by ROC curves plotted with the 'pROC' package. Immune infiltration was analyzed using CIBERSORT, and pan-cancer relationships were explored to identify associations between hub genes and various tumors. Potential therapeutic agents were investigated using the Drug Signatures Database (DSigDB). Experimental validation was conducted via RT-qPCR using cisplatin-induced chronic kidney disease (CKD) and ovariectomy (OVX)-induced osteoporosis models in C57BL/6J mice. After anesthesia and sacrifice, peripheral blood mononuclear cells (PBMCs) were collected to analyze the expression changes of hub genes. This study identified four hub genes (FAM184A, NFKBIA, RP2, HIRA). All hub genes exhibited excellent diagnostic performance, with FAM184A showing the best performance. Immune infiltration analysis revealed the relationships between hub gene expression levels and various immune cells. Pan-cancer analysis revealed the expression levels of FAM184A in different tumors, and it showed that high expression of FAM184A in SARC, SKCM, and PAAD is associated with improved prognosis and reduced mortality rates. Finally, RT-qPCR analysis revealed the mRNA expression levels of the hub genes in both OP and CKD. The mRNA expression of all hub genes were downregulated in osteoporosis model mice compared with normal mice, while in CKD mice, the mRNA expression of all hub genes except FAM184A was upregulated. This study identified four hub genes with significant diagnostic efficacy, suggesting they may act as crucial links between osteoporosis and chronic kidney disease. These genes offer promising targets for the treatment of both diseases. The findings of this study provide valuable insights for future research, which could further elucidate the complex pathogenetic mechanisms connecting chronic kidney disease and osteoporosis.