Positive Cells Research Articles

Pyridoxine supplementation before puberty ameliorates MAM-induced cognitive and sensorimotor gating impairments.

Schizophrenia is a kind of neurodevelopmental mental disorder in which patients begin to experience changes early in their development, typically manifesting around or after puberty and has a fluctuating course. Environmental disturbances during adolescence may be a risk factor for schizophrenia-like deficits. As a better treatment option, preventive intervention prior to schizophrenia may be more beneficial than direct treatment. More effective stress-relieving interventions during the critical puberty period may prevent schizophrenia-like neuronal changes and the transition to schizophrenia in adulthood. Pyridoxine deficiency alters the function of NMDA (n-methyl-D-aspartic acid) receptors and plays a key role in learning and memory. In this study, we prepared a progeny model of schizophrenia by exposing pregnant rats to methoxymethanol acetate (MAM) on gestational day 17. The offspring rats were injected intraperitoneally with pyridoxine daily from birth to prepuberty PND12-PND21), and behavioral changes in the offspring rats were observed in adulthood. Cannabinoid receptor interacting protein 1 (CNRIP1) and cannabinoid receptor-1 (CB1R), which regulate memory, cognitive and motor activity, were detected in the prefrontal cortex (PFC) and hippocampus of the offspring rats, and the cell proliferation in the hippocampal dentate gyrus (DG) was also observed. The results showed that the MAM rats spent less time the open arm in the elevated plus maze test, decreased discrimination coefficient in novel object recognition test, and decreased prepulse inhibition, while the MAM rats supplemented with pyridoxine in prepuberty did not show any of the above abnormal behavioral changes in adulthood. By examining related proteins in the PFC and hippocampus, we found that only CB1R protein expression was downregulated in the PFC, whereas CNRIP1 expression was not only elevated in the hippocampus, but also significantly increased in pyridoxine- supplemented MAM rats. Meanwhile, pyridoxine supplementation alleviated the reduction of doublecortin (DCX)-positive cells and Ki67-positive cells in MAM rats. These results indicate that prepuberty pyridoxine supplementation has a positive effect on the prevention of cognitive deficits and sensorimotor gating impairment in MAM-induced schizophrenia-like rats, accompanied by changes in the CB1R and CNRIP1 expression in PFC and hippocampus, as well as the regeneration of neurons in the DG region.

Novel Dynamic Organ Storage System Enhances Liver Graft Function in a Porcine Donation After Circulatory Death Model.

Donation after circulatory death (DCD) livers face increased risks of critical complications when preserved with static cold storage (SCS). Although machine perfusion (MP) may mitigate these risks, its cost and logistical complexity limit widespread application. We developed the Dynamic Organ Storage System (DOSS), which delivers oxygenated perfusate at 10°C with minimal electrical power requirement and allows real-time effluent sampling in a portable cooler. In a porcine DCD model, livers were preserved using DOSS or SCS for 10 hours and evaluated with 4 hours of normothermic MP, with n = 5 per group. After 4 hours of normothermic MP, the DOSS group demonstrated significantly lower perfusate lactate (p = 0.023), increased perfusate fibrinogen (p = 0.005), higher oxygen consumption (p = 0.018), greater bile production (p = 0.013), higher bile bicarbonate levels (p = 0.035) and bile/perfusate sodium ratio (p = 0.002), and lower hepatic arterial resistance after phenylephrine administration (p = 0.018). Histological analysis showed lower apoptotic markers in DOSS-preserved livers, with fewer cleaved caspase-3 (p = 0.039) and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL; p = 0.009) positive cells. These findings suggest that DOSS can enhance DCD allograft function during transport, offering potential clinical benefits and contributing to the expansion of the donor pool.

Infiltration of CD138-positive plasma cells in endoscopic material from patients with inflammatory bowel diseases

Introduction: CD138 (syndecan-1) expression hasbeen confirmed in plasma cells and normal tissues,particularly in epithelial cell types. As a cell surfaceprotein, CD138 regulates cell-cell and cell-matrixinteractions. CD138 may also regulate cellproliferation, migration, and cytoskeletal organiza-tion. In addition, positive CD138 plasma cells canalso be detected with extreme infiltration ininflammatory diseases such as chronic endometritis.Purpose: Investigation of the prognosticsignificance of the infiltration of CD 138 plasmacells in endoscopic material from patients withulcerative colitis (UC) and Crohn's disease (CD).Materials and methods: The study group included34 patients with ulcerative colitis and 8 patientsdiagnosed with CD. The expression of CD138 wasperformed by immunohistochemistry and assessedas a membrane-cytoplasmic color reaction inplasma cells localized in the lamina propria andglandular epithelium. We also examined theinsensitivity of general CD138-positive cells anddivided them into 4 groups: 0-absent, 1-mildincrease, 2-moderate increase, and 3-markedincrease.Results: The study group consists of 16 femalesand 18 males in UC and 2 female and 6 male ofCD. The UC lesions were mainly located in the rectum (19/34 cases), sigmoid (8/34cases), andothers (7/34 cases). Crohn's disease was observed inthe small intestine (3/8 cases), caecum (4/8 cases)and rectum (1/8 cases). The inflammatoryinfiltration consists of CD138+ cells in less than90% of inflammatory cells in UC (median 52.39%)and in less than 60% of inflammatory cells in CD(36.50%). The increase in CD138+ plasma cells inUC was mild in 3 cases, moderate in 10 cases andmarked in 15 cases. CD138+ plasma cells in UCwere found to correlate with Geboes score(R=0.404), CD15+ neutrophils (R=0.467) andCD15+macrophages (R=0.459). Plasma cells wereslightly increased in 2 cases and moderatelyincreased in 5 cases of CD. Infiltration of CD138+cells in CD sections was associated with age (R= -0.481) and crypt destruction (R=0.606). Loss ofCD138 expression in glandular epithelium wasdetected in 4/38 cases of UC and 1/8 cases of CD.Conclusion: Our results showed that infiltration ofCD138-positive plasma cells is closely related tothe degree of inflammation and architecturalchanges that could allow anti-CD138 treatment ininflammatory bowel disease.

Involvement of autophagy in germ cells in an experimental model of varicocele in rats before and after varicocelectomy.

Autophagy, a catabolic process enabling cellular organelles and proteins' reuse for energy, has been observed in varicocele models, but the effect of surgical treatment on this process remains unknown. This study aims to assess autophagy in varicocele models undergoing surgical correction. Twenty-one adolescent male rats were induced with varicocele and divided into three groups: sham, varicocele, and varicocele with varicocelectomy. After 21days, testicles were examined histologically for spermatogenesis (Jonhsen's score) and immunohistochemically for autophagy markers (LC3A, Beclin-1, Ambra-1, ULK-1, p62). Positive germ cells were quantitatively evaluated, and data were statistically analyzed (p < 0.05). Histological examination revealed significantly reduced Jonhsen's scores in varicocele compared to sham and varicocelectomy groups (p < 0.05). Expression of autophagy markers (LC3A, Beclin-1, Ambra-1, ULK-1, p62) was significantly higher in varicocele than sham and varicocelectomy groups (p < 0.05), and in varicocelectomy than sham (p < 0.05). Varicocele activates autophagy markers, with p62 potentially modulating autophagy despite being considered an inhibitor. While varicocelectomy improves histology, it doesn't fully inhibit autophagy, suggesting ongoing germ cell dysfunction despite treatment. This underscores varicocele's detrimental effects on germ cell functionality.

Early-life challenge enhances cortisol regulation in zebrafish larvae.

The hypothalamic-pituitary-adrenal (HPA) axis in mammals and the hypothalamic-pituitary-interrenal (HPI) axis in fish are open systems that adapt to the environment during development. Little is known about how this adaptation begins and regulates early stress responses. We used larval zebrafish to examine the impact of prolonged forced swimming at 5 days post-fertilization (dpf), termed early-life challenge (ELC), on cortisol responses, neuropeptide expression in the nucleus preopticus (NPO), and gene transcript levels. At 6 dpf, ELC-exposed larvae showed normal baseline cortisol but reduced reactivity to an initial stressor. Conversely, they showed increased reactivity to a second stressor within the 30-min refractory period, when cortisol responses are typically suppressed. ELC larvae had fewer corticotropin-releasing hormone (crh), arginine vasopressin (avp), and oxytocin (oxt)-positive cells in the NPO, with reduced crh and avp co-expression. Gene expression analysis revealed upregulation of genes related to cortisol metabolism (hsd11b2, cyp11c1), steroidogenesis (star), and stress modulation (crh, avp, oxt). These results suggest that early environmental challenge initiates adaptive plasticity in the HPI axis, tuning cortisol regulation to balance responsiveness and protection during repeated stress. Future studies should explore the broader physiological effects of prolonged forced swimming and its long-term impact on cortisol regulation and stress-related circuits.

Tissue-resident natural killer cells support survival in pancreatic cancer through promotion of cDC1-CD8 T activity.

The immunosuppressive microenvironment in pancreatic ductal adenocarcinoma (PDAC) prevents tumor control and strategies to restore anti-cancer immunity (i.e. by increasing CD8 T-cell activity) have had limited success. Here, we demonstrate how inducing localized physical damage using ionizing radiation (IR) unmasks the benefit of immunotherapy by increasing tissue-resident natural killer (trNK) cells that support CD8 T activity. Our data confirms that targeting mouse orthotopic PDAC tumors with IR together with CCR5 inhibition and PD1 blockade reduces E-cadherin positive tumor cells by recruiting a hypoactive NKG2D-ve NK population, phenotypically reminiscent of trNK cells, that supports CD8 T-cell involvement. We show an equivalent population in human single-cell RNA sequencing (scRNA-seq) PDAC cohorts that represents immunomodulatory trNK cells that could similarly support CD8 T-cell levels in a cDC1-dependent manner. Importantly, a trNK signature associates with survival in PDAC and other solid malignancies revealing a potential beneficial role for trNK in improving adaptive anti-tumor responses and supporting CCR5 inhibitor (CCR5i)/αPD1 and IR-induced damage as a novel therapeutic approach.

Pyroptosis was suppressed by 20-hydroxyecdysone through Lin28b-mediated let-7d in vascular endothelial cells.

20-hydroxyecdysone (20E), a natural polyhydroxylated steroid, has exhibited anti-inflammatory effects across various diseases. This study investigates the potential connection between 20E's anti-inflammatory properties and the RNA-binding protein Lin28b, which is notably upregulated in TNF-α-stimulated endothelial cells. Herein, we discovered that 20E can selectively downregulate Lin28b expression without affecting its paralog Lin28a. Notably, 20E treatment could significantly attenuate pyroptosis, an inflammatory form of programmed cell death, as evidenced by reduced IL-1β and LDH release, and fewer propidium iodide (PI)-positive cells. Subsequent protein analysis revealed that 20E inhibited the enhanced expressions of key pyroptosis-associated proteins, GSDMD, GSDMD-N, and GSDME-N. Besides, this suppression of Lin28b and pyroptosis may be partially mediated through TNFR1. Additionally, 20E upregulated let-7 miRNA, particularly let-7d, a critical downstream target of Lin28b. To elucidate the role of Lin28b in 20E-mediated pyroptosis attenuation, we performed Lin28b overexpression and silencing experiments. Overexpressing Lin28b negated 20E's inhibition of LDH release and PI-related fluorescence, exacerbating GSDMD and GSDME cleavage. Conversely, Lin28b knockdown augmented the suppressive effect of 20E on pyroptosis, which was reversed by a let-7d inhibitor. Co-transfection with let-7d mimic and Lin28b plasmid demonstrated let-7d's role in mitigating pyroptosis aggravated by Lin28b overexpression. Overall, this study demonstrates that 20E may mitigate GSDMD and GSDME-mediated pyroptosis in HUVECs through the Lin28b/let-7d-dependent signaling pathway. These results highlight the potential of 20E as a promising inhibitor of pyroptosis, offering new insights into its anti-inflammatory mechanisms.

Case report: Vitamin B12 deficiency-associated hemolytic anemia

Vitamin B12 deficiency is commonly known to cause hematological, neurological, and gastrointestinal disturbances including anemia, pancytopenia, sub-acute combined degeneration, and glossitis, among others. Hemolytic anemia is a rare but possible presentation of vitamin B12 deficiency. We present a case series of three elderly females with different past medical histories but with similar presentations of severe anemia. Initial investigation revealed pancytopenia and hemolytic anemia. Further evaluation indicated vitamin B12 deficiency and subsequently, pernicious anemia, as confirmed by low serum vitamin B12 levels and positive intrinsic cell antibody respectively. These findings directed the diagnosis toward vitamin B12 deficiency-induced hemolytic anemia, a rare clinical entity. All three patients responded to treatment with intramuscular vitamin B12 injections, showing significant improvement in their pancytopenia and hemolytic anemia. This case series highlight the importance of considering vitamin B12 deficiency as a potential cause of hemolytic anemia in patients presenting with pancytopenia and ineffective hematopoiesis. Further research is necessary to illuminate the therapeutic implications of this rare condition.

Histomorphometric analysis of external apical root resorption using L-loops and different intrusive forces

ABSTRACT Purpose This study aimed to conduct a histomorphometric analysis on the impact of varying intrusive forces on external apical root resorption (EARR) in rat molars, using an L-loop was employed to apply intrusion forces, artificially inducing EARR in the maxillary left first molar (M1) of rats. Materials and methods The L-loop was bent on a plaster rat model using stainless steel round wires with diameters of 0.014 and 0.016 inches. Using a universal test force measuring device, L-loops were calibrated to 4 N/mm and 6 N/mm for the 0.014-inch and 0.016-inch diameter stainless steel round wires, respectively. Fifteen-week-old Sprague-Dawley (SD) male rats were used and divided into the control (no loop) and experimental (0.014 and 0.016) groups (n = 5 for each group). The L-loop was attached to the left maxillary molar, applying an intrusive force to M1. After 15 days, the rats were euthanized. Maxillary bones were analysed using micro-computed tomography (μCT), evaluating M1 intrusive distance of vertical tooth movement and counting the number of tartrate-resistant acid phosphatase (TRAP)-positive cells on the surface of the mesial root apex and alveolar bone. Results The L-loop groups exhibited significantly increased M1 intrusive distance and TRAP-positive cell counts compared to controls. Specifically, the 0.016 group demonstrated greater M1 intrusive distance and more TRAP-positive cells than the 0.014 group. The M1 intrusive distance and TRAP-positive cells increased when intrusive forces were applied using L-loops of different diameters. Conclusion Greater intrusive forces enhance TRAP-positive cell activity, influencing EARR and vertical tooth movement.

Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro Culture

Rat dental pulp stem cells (DPSCs) can be used to elucidate mesenchymal stem cell (MSC) applications in regenerative medicine. However, information on rat DPSCs during long-term passage, which could lead to replicative senescence, is limited. In this study, we investigated the phenotypic changes in DPSCs after 3–26 passages (3P–26P). The results show that cell morphology and nuclear size increase proportionally with passage number. The phosphorylated histone H2A.X (γ-H2A.X) positive cells (indicating DNA damage) increased significantly earlier than the 4-Hydroxynonenal (4-HNE) stained cells (indicating an abundance of intracellular reactive oxygen species). Compared to the cells subjected to 3P and 5P, the cells subjected to 15P showed reduced proliferation despite being positive for Ki67. Furthermore, cell growth was completely arrested after 26P. The senescence markers, senescence-associated β-galactosidase (SA-β-gal) and p16, exhibited similar expression patterns that were not correlated with those of p21 and urokinase-type plasminogen activator receptor (uPAR). Nearly all cells expressed SA-β-gal and p16 after 26P, whereas only half expressed p21 and uPAR. These results will contribute to understanding the characteristics of DPSCs toward replicative senescence, which are applicable to elucidate mechanisms related to regenerative medicine and stem cell aging.

Adjuvant Radiation Sparing after Neoadjuvant Chemotherapy and TORS in Selected HPV-Positive Oropharyngeal Cancer.

Transoral robotic surgery (TORS) has shown promising results in treating human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC), and there has been increasing interest in incorporating neoadjuvant chemotherapy (NCT) prior to TORS. This study aimed to assess the feasibility and safety of sparing adjuvant RT following NCT and TORS. A retrospective cohort study included consecutive patients with HPV-positive OPSCC who underwent NCT followed by TORS without adjuvant RT. Disease-free survival (DFS) was the primary outcome. Pattern of recurrence (local, regional, and distant), salvage treatment outcomes, and predictors of tumor recurrence were also assessed. A total of 84 patients were included in the analysis. No patients died during the study period. DFS rates (95% Confidence Interval, CI) at 1, 2, and 3 years were 92.8% (87.4-98.5), 87.0% (79.7-94.9), and 84.4% (76.0-93.8), respectively. Local, regional, and distant recurrence rates were 7.0%, 9.5%, and 3.6%, respectively. Salvage treatment achieved a 100% salvage rate. Predictors of tumor recurrence included the number of positive lymph nodes (hazard ratio: 2.66; 95% CI: 1.19-5.92) and clinical stage III at diagnosis (hazard ratio: 7.65; 95% CI: 1.97-29.7). Recommendation of adjuvant treatment based on pathologic adverse features appears to be associated with favorable outcomes in selected HPV-positive OPSCC cases treated with NCT and TORS. Future studies should focus on refining criteria for recommending adjuvant RT to further reduce recurrence rates and minimize treatment-related toxicity, contributing to personalized treatment strategies for HPV-related OPSCC. 4 Laryngoscope, 2024.

Perifollicular concentric granulomas: A clue to IgG4-related lymphadenopathy.

A 69-year-old with well-controlled HIV was evaluated for persistentcough, in the context of years of fatigue and influenza A infection 6months prior. Chest CT and PET scans were notable for adenopathyconcerning for a lymphoproliferative disorder. Radiologic studies alsoshowed diffuse FDG uptake in the prostate, consistent with prostatitis.Axillary lymph node biopsy showed follicular and paracorticalhyperplasia, and few germinal centers showed perifollicular non-necrotizing granulomas. Immunohistochemical staining demonstrated apredominance of IgG4 positive plasma cells. Serum proteinelectrophoresis (SPEP) and immunosubtraction showed a board-domedpeak pattern suggestive of possible monoclonality. Serum IgG4 levelswere elevated, and the patient was diagnosed with IgG4-relateddisease (IgG4-RD). This case highlights morphologic and SPEP patternsthat can aid in supporting a diagnosis of IgG4-RD.

Expression of CD68+ Cells in Synovial Tissue from Patients with PsA and its Association with Disease Activity Indices: A Clinical Pilot Study.

Investigating CD68+ positive cells in the synovial tissue is crucial for understanding the pathogenesis of psoriatic arthritis (PsA) and developing targeted treatment strategies. The role of CD68+ positive cells in the synovial tissue of patients with PsA for joint destruction has not been fully studied. The objective of the study was to examine the presence of CD68+ cells in the synovial tissue of patients with PsA, particularly those with high inflammatory activity. Synovial tissue samples were collected during knee joint replacement surgeries from patients with PsA (16 patients) and gonarthrosis (25 patients). Immunohistochemical methods were employed to detect CD68+ cell expression in the tissue samples. The results were analyzed by histologists, and the staining intensity and percentage of positively stained cells were evaluated. The data were then divided into three groups for statistical analysis: negative, weakly positive, and strongly positive histological samples. Routine indices for disease activity, VAS, DAPSA, PASDAI, and mCPDAI were used to assess PsA activity in all patients and to assess correlations with CD68+ positive cells in the synovial tissue. Statistical analysis was performed using SPSS version 26.0 (SPSS Inc., Chicago, IL, USA). The expression of CD68+ positive cells was significantly higher in patients with PsA compared to those with activated gonarthrosis (p < 0.001). The indices for disease activity, VAS, DAPSA, PASDAI, mCPDAI, and mCPDAI showed a significant positive relationship with the expression of CD68 + cells on synovial tissue in patients with PsA (p < 0.01) Conclusion: The findings of the study confirm the increased numbers of CD68+ cells in PsA vs. gonathrosis synovium. This suggests the need to explore therapeutic approaches aimed at suppressing or blocking CD68+ cells to potentially mitigate joint damage.

ROS-Induced Gingival Fibroblast Senescence: Implications in Exacerbating Inflammatory Responses in Periodontal Disease.

Periodontal disease is the pathological outcome of the overwhelming inflammation in periodontal tissue. Cellular senescence has been associated with chronic inflammation in several diseases. However, the role of cellular senescence in the pathogenesis of periodontal disease remained unclear. This study aimed to investigate the role and the mechanism of cellular senescence in periodontal disease. Using single-cell RNA sequencing, we first found the upregulated level of cellular senescence in fibroblasts and endothelial cells from inflamed gingival tissue. Subsequently, human gingival fibroblasts isolated from healthy and inflamed gingival tissues were labeled as H-GFs and I-GFs, respectively. Compared to H-GFs, I-GFs exhibited a distinct cellular senescence phenotype, including an increased proportion of senescence-associated β-galactosidase (SA-β-gal) positive cells, enlarged cell morphology, and significant upregulation of p16INK4A expression. We further observed increased cellular reactive oxygen species (ROS) activity, mitochondrial ROS, and DNA damage of I-GFs. These phenotypes could be reversed by ROS scavenger NAC, which suggested the cause of cellular senescence in I-GFs. The migration and proliferation assay showed the decreased activity of I-GFs while the gene expression of senescence-associated secretory phenotype (SASP) factors such as IL-1β, IL-6, TGF-β, and IL-8 was all significantly increased. Finally, we found that supernatants of I-GF culture induced more neutrophil extracellular trap (NET) formation and drove macrophage polarization toward the CD86-positive M1 pro-inflammatory phenotype. Altogether, our findings implicate that, in the inflamed gingiva, human gingival fibroblasts acquire a senescent phenotype due to oxidative stress-induced DNA and mitochondrial damage, which in turn activate neutrophils and macrophages through the secretion of SASP factors.

M1 polarization of hepatic macrophages in cows with subclinical ketosis is an important cause of liver injury.

Subclinical ketosis (SCK) is highly prevalent and easily overlooked, with insidious and slow progression of hepatic injury, often characterized by an imbalance in immune homeostasis. In nonruminants, macrophage polarization plays an important regulatory role in hepatic lipid accumulation, fibrosis and inflammatory processes. Thus, we aimed to investigate the status of hepatic macrophage polarization in SCK cows and to corroborate its association with liver injury and inflammation. Twelve Holstein dairy cows (parity, 2-4) were selected, and liver biopsy and blood were collected on the second week postpartum (10-14 d days in milk). On the basis of serum BHBA concentrations., selected cows were categorized into healthy (n = 6; BHBA <1.0 mM) and SCK (n = 6; 1.2 mM ≤ BHBA < 3.0 mM) groups. Serum biochemical parameters were measured using an automatic biochemical analyzer, which indicated higher serum levels of BHBA and NEFA and an upregulation of liver injury indicators (AST, ALT, TP, GLB) in SCK cows compared with healthy cows. ELISA assays revealed that SCK cows displayed systemic low-grade inflammation, as demonstrated by increased serum levels of HP, SAA, TGF-β, IFN-γ, and IL-1β. Liver biopsies revealed pathological histological alterations, hepatic inflammation, and macrophage polarization status. Oil red staining indicated steatosis, while Sirius red staining demonstrated mild extracellular matrix deposition in the liver of SCK cows. The expression of inflammatory response-related proteins (TLR4, p-NFκB, p-I-κB, NLRP3, and Caspase-1) was elevated in the liver of SCK cows, with the increased mean fluorescence intensity of NFκB further confirming the activation of the inflammatory pathway. Furthermore, the levels of pro-inflammatory factors, TNF-α and IFN-γ, were elevated in the tissue homogenate. Macrophage phenotypic changes in SCK cows were further explored based on the results of liver injury and inflammation. Compared with healthy cows, the protein and mRNA abundance of the macrophage marker CD68 in the liver of SCK cows was higher, along with an increased mean fluorescence intensity of CD68. SCK cows also exhibited reduced mRNA expression of the Kupffer cell marker CLEC4F and elevated chemokine levels (CXCL1 and CCL2). As evidenced by greater protein and mRNA abundance of macrophage M1 polarization markers (iNOS, IL-1β, CD86, IL-6, IL-12b, and CCL3), higher fluorescence intensity of iNOS and CD86, and an increased number of CD68+/CD86+-positive cells observed via immunofluorescence, the macrophage polarization phenotype in the liver of SCK cows was predominantly M1. In contrast, the protein and mRNA abundances of M2 polarization markers (CD206, IL-10, and Arg1) were lower in SCK cows, accompanied by a reduced fluorescence intensity of CD206 and a lower number of CD68+/CD206+-positive cells. Overall, the present study revealed that the number of macrophages in liver is enhanced during subclinical ketosis and is dominated by pro-inflammatory macrophages (M1 macrophages). This could partly explain the increased risk of steatosis, fibrosis, and inflammatory response processes in these cows.

Characterization and Biocontrol Efficacy of Bacillus velezensis GYUN-1190 against Apple Bitter Rot.

The application of synthetic fungicides has resulted in environmental pollution and adverse effects on non-target species. To reduce the use of agrochemicals, crop disease management requires microbial biological control agents. Bacillus-related genera produce secondary metabolites to control fungal pathogens. Bacillus velezensis GYUN-1190, isolated from soil, showed antagonistic activity against Colletotrichum fructicola, the apple anthracnose pathogen. Volatile organic compounds and culture filtrate (CF) from GYUN-1190 inhibited C. fructicola growth in vitro, by 80.9% and 30.25%, respectively. The CF of GYUN-1190 inhibited pathogen spore germination more than cell suspensions at 10 8 cfu/ml. Furthermore, GYUN-1190 CF is effective in inhibiting C. fructicola mycelial growth in vitro, and it suppresses apple fruit bitter rot more effectively than GYUN-1190 cell suspensions and pyraclostrobin in planta. The mycelial growth of C. fructicola was completely inhibited 48 h after immersion into the CF, in compared with positive controls and GYUN-1190 cell suspensions. The genetic mechanism underlying the biocontrol features of GYUN-1190 was defined using its whole-genome sequence, which was closely compared to similar strains. It consisted of 4,240,653 bp with 45.9% GC content, with 4,142 coding sequences, 87 tRNA, and 28 rRNA genes. The genomic investigation found 14 putative secondary metabolite biosynthetic gene clusters. The investigation suggests that B. velezensis GYUN-1190 might be more effective than chemical fungicides and could address its potential as a biological control agent.

Identification of new antigenic epitopes of porcine reproductive and respiratory syndrome virus nsp12 protein using monoclonal antibodies

Porcine reproductive and respiratory syndrome virus (PRRSV), a member of the arteritis virus family, significantly impacts the swine industry due to its high infectiousness. nsp12, the nonstructural protein encoded by PRRSV, is a membrane-associated protein, with limited knowledge about its antigenic properties and functions. In this study, we used the expressed and purified nsp12 protein as antigen to immunize mice, and successfully screened five positive hybridoma cell lines that stably secrete anti-nsp12 monoclonal antibodies using immunological assays such as indirect ELISA and IFA. The antigenic epitopes recognized by the five monoclonal antibodies were identified using the fusion expression of peptides derived from the overlapping truncators of the nsp12 gene. The results showed that monoclonal antibodies 3G11 and 9C2 recognized the antigenic epitope 93TWGFESDTAY102, 2E3 recognized 115DYNDAFRARQ124, and 10G6 and 2A4 recognized 142PGPVIEPTL150. Furthermore, the three newly identified antigenic epitopes were all immunodominant and located on the surface of nsp12 protein. Notably, the antigenic epitopes 93–102 aa are all highly conserved across PRRSV-2 strains, making them suitable targets for PRRSV-2 detection. In conclusion, our findings advance the understanding of the antigenic properties of the PRRSV nsp12 protein and facilitate the development of assays for PRRSV detection.

Exposure to an environmentally representative mixture of polybrominated diphenyl ethers (PBDEs) alters zebrafish neuromuscular development

Polybrominated diphenyl ethers (PBDEs) are a prevalent group of brominated flame retardants (BFRs) added to several products such as electronics, plastics, and textiles to reduce their flammability. They are reported as endocrine disruptors and neurodevelopmental toxicants that can accumulate in human and wildlife tissues, thus making their ability to leach out of products into the environment a great cause for concern. In this study, zebrafish (Danio rerio) embryos and larvae were exposed to a wide concentration range (1.5, 15, 150 and 300 pM) of a PBDE mixture from one to six days post-fertilization (dpf). Hatching rates, mortality and general morphology were assessed during the exposure period. A delay in hatching was observed at the two highest PBDEs concentrations and mortality rate increased at 6 dpf. By 4 dpf, larvae exposed to 150 pM and 300 pM PBDEs developed an upcurved phenotype. Analysis of motor behavior at 6 dpf revealed that PBDE exposure acutely reduced locomotion. To further analyze these motor deficits, we assessed the neural network density and motor neuron and neuromuscular junctions (NMJ) development by immunostaining and imaging. Acetylated α-tubulin staining revealed a significant loss of neurons in a dose-dependent manner. Synaptic vesicle protein 2 (SV2) and ⍺-bungarotoxin (⍺-BTX) staining revealed a similar pattern, with a significant loss of SV2 and nicotinic acetylcholine receptors, thus preventing the colocalization of presynaptic neurons with postsynaptic neurons. Consistent with these results, the presence of cleaved caspase-3 and acridine orange positive cells showed increased cell death in zebrafish larvae exposed to PBDEs. Our results suggest that exposure to PBDEs leads to deficits in the zebrafish neuromuscular system through neuron death, inducing morphological and motor deficiencies throughout their development. They provide valuable insight into the neurotoxic effects of PBDEs, further highlighting the relevance of the zebrafish model in toxicological studies.

Neuroprotective effects of biogenic silver nanoparticles in Parkinson's disease: In vitro and in vivo study

The study aimed to develop immunomodulatory silver nanoparticles (CHT-PCT/AgNPs) using a green chemistry approach with Umbilicaria esculenta (U.E.) extract to protect neurons from Parkinson's disease. The synthesized CHT-PCT/AgNPs measured 88.2 ± 33.7 nm in size, with a zeta potential of approximately 24.5 ± 1.9 mV. In vitro assays demonstrated that the green-synthesized nanoparticles exhibited greater biocompatibility and antioxidant properties compared to chemically synthesized AgNPs. The MTT assay indicated that CHT-PCT/AgNPs were cytocompatible up to 50 μg/mL, but higher concentrations resulted in significant toxicity. The DPPH assay revealed dose-dependent antioxidant activity, although it was lower than that of ascorbic acid. In animal studies of MPTP-induced Parkinsonism, the count of tyrosine hydroxylase (TH)-positive cells varied across groups, with healthy controls showing 145.3 ± 11.0 cells compared to the negative control group with only 21.6 ± 4.1. Additionally, the number of DRD1-positive cells also reflected a similar trend. These findings suggest that CHT-PCT/AgNPs may serve as a potential neuroprotective agent to mitigate Parkinson's disease.

Pleiotrophin Overexpression Reduces Adolescent Ethanol Consumption and Modulates Ethanol-Induced Glial Responses and Changes in the Perineuronal Nets in the Mouse Hippocampus.

To investigate whether pleiotrophin (PTN) overexpression influences ethanol consumption during adolescence and its effects on glial responses, neurogenesis, and perineuronal nets (PNNs) in the mouse hippocampus. Male and female adolescent transgenic mice with elevated PTN levels (Ptn-Tg) and controls underwent an intermittent access to ethanol (IAE) 2-bottle choice protocol. Ethanol consumption, PTN levels, neurogenesis, and glial responses were measured in the hippocampus. Immunohistochemistry was used to assess changes in new neurons, microglial and astrocyte populations, and PNNs. Ptn-Tg mice consumed significantly less ethanol compared to controls, irrespective of sex. Chronic alcohol exposure reduced PTN levels in the hippocampus. PTN overexpression decreased the number of new neurons in the dentate gyrus (DG) and prevented ethanol-induced microglial activation. Ptn-Tg mice had significantly more astrocytes and fewer PNNs, with a higher percentage of parvalbumin (PV) positive cells surrounded by PNNs under basal conditions. However, ethanol drastically reduced the number of PV+ cells in the DG of Ptn-Tg mice, despite the presence of PNNs. PTN overexpression reduces adolescent ethanol consumption and influences ethanol-induced effects on hippocampal neurogenesis, glial responses, and PNN remodeling. These findings underscore the importance of PTN in modulating alcohol-induced neurotoxicity.