Search Results (714)

Background/Objectives: In case of EDTA-induced pseudothrombocytopenia (PTCP), MgSO₄-anticoagulated tubes are recommended for platelet counting, requiring the collection of an additional tube. The aim of this study was to analyze whether complete blood count (CBC) and differential performed on MgSO₄-anticoagulated tubes were comparable to the results obtained on K₃-EDTA samples, and to characterize the stability of the CBC over a 24 h period. Methods: In 355 patients (70 with a confirmed PTCP and 285 without PTCP), we compared CBC results obtained on K₃-EDTA- and MgSO₄-anticoagulated tubes, using DxH800 analyzers. In 33 cases, a differential was available for both anticoagulants, and for 10 patients, samples were re-analyzed 6, 12, and 24 h after the first determination. Results: In the presence or absence of clumps, white blood cell (WBC) count, hematocrit, and mean corpuscular volume (MCV) were slightly lower in MgSO₄ than in K₃-EDTA tubes, whereas mean corpuscular hemoglobin concentration (MCHC) was slightly higher. Mean platelet volume (MPV) was significantly lower on MgSO₄- than on K₃-EDTA-anticoagulated tubes. Values were highly correlated between both anticoagulants, and mean relative biases (MRBs) were below Ricos’s recommendations, except for MCHC and MPV. For differential, neutrophils were significantly lower on MgSO₄- in comparison to K₃-EDTA-anticoagulated tubes (MRB = −2.9%, below Ricos’s optimal bias). The morphology of white blood cells (WBCs) was similar on both anticoagulants. During storage at room temperature, MCV and red cell distribution width increased slightly, but the increase was more pronounced in K₃-EDTA than in MgSO₄ tubes. Conclusions: CBC and differentials obtained with the DxH 800 analyzer on MgSO₄-anticoagulated samples are similar to those obtained with K₃-EDTA, except for MPV. Full article

Plant-based symbiotic systems are often limited by poor storage stability and inconsistent biofunctional performance. This study evaluated the stability and functionality of a fermented blend based on sacha inchi (Plukenetia volubilis) oil press cake (SIC) and yacon (Smallanthus sonchifolius) flour (YF) as sources of protein and fructooligosaccharides (FOS), respectively, using two processing strategies: fermentation with Lactobacillus rhamnosus (T1) and combined enzymatic hydrolysis with Alcalase and fermentation with Lactobacillus plantarum (T2). Both treatments maintained viable cell counts (VCC) above probiotic thresholds (>10⁶ CFU mL⁻¹) during 28 days of storage at 4 °C, confirming their suitability as probiotic carriers. Notably, T2 significantly enhanced metabolic activity, as evidenced by higher organic acid production and increased soluble protein content due to Alcalase-mediated hydrolysis, which promoted the generation of bioactive peptides associated with improved antioxidant and antihypertensive activities. Biofunctional properties, including total phenolic content, antioxidant capacity (AC), and angiotensin-converting enzyme (ACE) inhibitory activity, remained stable throughout storage, while FOS degradation was minimal, confirming preservation of prebiotic functionality. LC–MS/MS Q-TOF analysis revealed a complex phenolic profile that was differentially modulated by lactic acid fermentation, with L. plantarum (T2) promoting extensive phenolic biotransformation and increased metabolite diversity, whereas L. rhamnosus (T1) largely preserved the original phenolic profile. These findings demonstrate that the synergistic interaction between enzymatic hydrolysis and L. plantarum fermentation promoted peptide release, intensified microbial metabolism, and enhanced phenolic biotransformation, thereby contributing to the superior functional properties observed in T2, while maintaining stable biofunctional characteristics throughout refrigerated storage in both treatments. Full article

Background: Virological suppression is a key outcome of antiretroviral therapy. Despite progress in HIV treatment in Kazakhstan, virological non-suppression remains a relevant clinical and public health issue requiring further analysis. This study aimed to assess the prevalence of virological suppression (VS) and to identify factors associated with the absence of VS among people living with human immunodeficiency virus (HIV) who are receiving antiretroviral therapy (ART) in Kazakhstan. Methods: A retrospective cross-sectional analytical study was conducted using secondary analysis of a de-identified national registry database of people living with HIV (PLHIV) receiving ART in the Republic of Kazakhstan as of 30 September 2025. The primary outcome was virological non-suppression (VNS), defined as the last viral load (VL) value of at least 200 copies per milliliter. The analysis included sex, age, presumed route of HIV transmission, the first available cluster of differentiation 4 (CD4) cell count recorded in the registry, the last recorded percentage category of adherence to ART, and the aggregated category of ART regimen. The main descriptive, bivariate, and multivariable analyses were performed using a complete-case approach. Independent associations were assessed using multivariable logistic regression, and the results were presented as adjusted odds ratios (aORs) with 95 percent confidence intervals (CIs). Results: The initial registry extraction included 33,614 records, of which 32,130 patients were included in the final analytical sample. VS was achieved in 29,454 (91.7%) patients, whereas VNS was observed in 2676 (8.3%) patients. In the multivariable model, higher adjusted odds of VNS were observed among men compared with women (aOR 1.14; 95% CI 1.02–1.26), as well as among patients with a first CD4 count < 200 cells/μL compared with those with a first CD4 count of ≥500 cells/μL (aOR 1.25; 95% CI 1.09–1.44). The strongest association was found for reduced adherence to therapy. Compared with adherence of at least 95%, the adjusted odds of VNS were markedly higher among patients with adherence of 85–94% (aOR 28.66; 95% CI 25.85–31.77) and among those with adherence below 85% (aOR 61.05; 95% CI 50.50–73.81). In all age groups older than 25 years, the adjusted odds of VNS were lower than among patients younger than 25 years. Lower adjusted odds of VNS were also observed among patients with homosexual transmission, vertical transmission, and other or unspecified transmission routes compared with heterosexual transmission. Among ART regimens, regimens containing non-nucleoside reverse transcriptase inhibitors (NNRTIs) were associated with lower adjusted odds of VNS than dolutegravir-containing regimens (DTG-containing regimens) (aOR 0.68; 95% CI 0.52–0.88), whereas no statistically significant differences were identified for regimens containing protease inhibitors (PIs). Conclusions: Despite the high overall level of VS among PLHIV receiving ART in Kazakhstan, VNS remains concentrated in clinically and programmatically important subgroups. It was most strongly associated with reduced adherence and was also associated with younger age, marked baseline immunosuppression, and male sex in the primary model. These findings support the need for targeted interventions focused on adherence support, early diagnosis, and differentiated long-term follow-up of patients. Full article

Selective dry cow therapy (SDCT) has emerged as a key strategy to reduce antimicrobial use in dairy production while maintaining udder health. This study aimed to evaluate the feasibility and impact of implementing SDCT in Romanian dairy farms by comparing low-risk cows treated with internal teat sealant only (ITS) at dry-off with low-risk cows treated with intramammary antibiotics at dry-off. A prospective field study was conducted on two commercial dairy herds, including 87 cows classified based on somatic cell count (SCC) and differential SCC (DSCC), and compared with a historical cohort of 37 cows. Udder health parameters, milk yield during the first 100 days in milk (DIM), antimicrobial use, and economic outcomes were evaluated. No significant differences were observed between groups in terms of postpartum intramammary infections, somatic cell score, DSCC, or clinical mastitis incidence. Milk yield during early lactation was also not affected by treatment. The ITS-only strategy resulted in a substantial reduction in antimicrobial use (−88.8% per cow) without significant differences in total economic costs. Farm-related differences highlighted the influence of management conditions on outcomes. These findings indicate that, in low-risk cows, SDCT using ITS alone is a safe and effective alternative to antibiotic treatment and support the feasibility of implementing SDCT under Romanian dairy production conditions as a sustainable strategy to promote the targeted and prudent use of antimicrobials while reducing unnecessary antibiotic exposure in dairy herds. However, given the limited number of herds and animals included, further studies are needed to confirm these promising findings under a broader range of production conditions. Full article

Muscovy ducks (Cairina moschata) exhibit strong nesting tendencies, which result in reduced egg-laying performance. The research team previously identified differential expression of the Ras homolog family member B (RHOB) gene in the ovaries of Muscovy duck during the nesting and laying periods through RNA-seq and quantitative real-time polymerase chain reaction (qPCR) analysis. This finding suggested that RHOB may be associated with nesting behavior in Muscovy ducks. Previous studies have demonstrated that the nesting behavior of Muscovy ducks is closely associated with the proliferation and apoptosis of their ovarian granulosa cells. It is speculated that RHOB may be involved in the proliferation and apoptosis of Muscovy duck ovarian granulosa cells. This study employed qPCR, immunofluorescence staining, live-cell Caspase3 activity and mitochondrial membrane potential assays, reactive oxygen species (ROS) staining, 5-Ethynyl-2′-deoxyuridine (EdU) staining, cell cycle analysis, cell apoptosis detection and cell counting kit-8 (CCK-8) assays. Our results showed that RHOB inhibited granulosa cell apoptosis and promoted granulosa cell proliferation. Similarly, in a granulosa cell apoptosis model, RHOB was also found to inhibit apoptosis in Muscovy duck granulosa cells. Further studies revealed that RHOB regulates mitochondrial function in granulosa cells. The combined experimental results indicate that RHOB regulates granulosa cell apoptosis in Muscovy duck follicles via the mitochondrial apoptosis pathway. These findings provide an experimental basis and theoretical foundation for the selective breeding of desirable traits in Muscovy ducks, such as low nesting behavior and high egg production. Full article

Background: Remodeling of epithelial junctional architecture contributes to colorectal cancer (CRC) progression; however, the spatial organization linking tight-junction components to early dissemination remains incompletely characterized. Claudin-2 (CLDN-2) is frequently upregulated in CRC, yet whether it is associated with compartment-specific epithelial remodeling has not been systematically examined. Methods: In a retrospective single-center cohort of 54 surgically resected CRCs, we integrated clinicopathological variables, quantitative tumor budding counts, compartment-specific membranous E-cadherin expression, lymphovascular invasion, lymphoid follicles, and immune-cell densities. Analyses focused on spatial structural relationships within the tumor. Results: Higher CLDN-2 expression was enriched among node-positive tumors and advanced TNM stages. CLDN-2–higher tumors exhibited increased tumor budding and spatially selective adhesion remodeling, characterized by reduced membranous E–cadherin at the invasive front and budding sites, with more preserved membranous epithelial organization within metastatic lymph-node deposits. Descriptive co-occurrence and correlation analyses demonstrated concordant spatial relationships among CLDN-2 expression, tumor budding, nodal involvement, lymphovascular invasion, and compartment-specific E-cadherin patterns. In contrast, immune-related parameters showed weaker differentiation across CLDN-2 strata. Conclusions: CLDN-2 expression is associated with spatial epithelial remodeling in colorectal cancer, characterized by compartment-specific adhesion changes and increased microinvasive activity. The findings support a model in which CLDN-2 expression aligns with an invasion-associated epithelial configuration linked to tumor budding and nodal dissemination. These observations warrant validation in independent cohorts with outcome data. Full article

Pembrolizumab is standard adjuvant therapy for high-risk clear cell renal cell carcinoma, but serositis is an uncommon immune-related adverse event that may mimic recurrence or infection. We report a 55-year-old man who achieved no evidence of disease after nephrectomy and metastasectomy and developed anasarca, large bilateral pleural effusions, mild ascites, peripheral eosinophilia, and a small pericardial effusion after six cycles of adjuvant pembrolizumab. Pleural fluid was exudative and contained 20% eosinophils. Cytology showed inflammatory cells without evidence of malignancy; bacterial, mycobacterial, and fungal studies were negative; and mildly elevated adenosine deaminase did not support tuberculosis. Cardiac function and natriuretic peptides were preserved. Pembrolizumab was discontinued, thoracentesis and corticosteroids were administered, and symptoms, eosinophilia, renal function, and albumin improved rapidly. Follow-up through March 2026 showed no oncologic progression, although some residual pleural and abdominal fluid persisted alongside imaging findings suggestive of portal-hypertension physiology, which may have contributed to residual fluid but did not explain the eosinophilic pleural syndrome. In a targeted literature review, effusion eosinophil data were infrequently reported. This case highlights a likely underrecognized eosinophilic pleural-fluid phenotype within pembrolizumab-associated polyserositis and supports routine differential cell counts in drained serosal fluid when immune-related serositis is suspected. Full article

Bovine mastitis remains a major impediment to optimal dairy production. Somatic cell count (SCC) is commonly used as an indicator of mammary gland inflammation, while milk microbiota may also reflect mastitis-related changes. Here, we employed Oxford Nanopore full-length transcript sequencing to delineate the peripheral blood transcriptomic landscape of Xinjiang Brown cattle stratified by high (SCC ≥ 1,000,000 cells mL⁻¹) and low (SCC ≤ 200,000 cells mL⁻¹) SCCs, with the objective of identifying candidate genes underpinning mastitis resistance. We identified 226 differentially expressed genes and 441 differentially expressed transcripts. Genes in the high-SCC group were prominently enriched in immune response pathways and chemokine signalling cascades. Protein–protein interaction network analysis further delineated a core module of ten immune-related genes, including CCL4, IL1B and CXCL2. Integrative analysis with complementary second-generation sequencing data pinpointed CXCL2 as a high-priority candidate. Subsequent RT–qPCR and enzyme-linked immunosorbent assay (ELISA) validation revealed that CXCL2 expression was significantly elevated both in high-SCC individuals and in an LPS-induced bovine mammary epithelial cell inflammation model. Collectively, these findings establish CXCL2 as a putative molecular marker for mastitis resistance breeding and provide a foundational resource for deciphering the molecular mechanisms governing mammary health. Full article

Background/Objectives: Early functional status at hospital discharge is a clinically relevant outcome after aneurysmal subarachnoid hemorrhage (aSAH), but early prognostic assessment remains challenging. We evaluated whether admission inflammatory blood cell ratios were associated with discharge independence and added prognostic information beyond established neurological severity scales. Methods: In this retrospective single-center cohort study, 252 consecutive adults with aSAH were screened, and 144 endovascularly treated patients with available admission complete blood count with differential were included. Discharge independence was defined as a Barthel Index score ≥60 at hospital discharge. A clinical reference model included age, World Federation of Neurosurgical Societies (WFNS) grade, and Hunt–Hess grade. Multivariable logistic regression was used to assess associations between inflammatory ratios and discharge independence. Discrimination was assessed using receiver operating characteristic analysis with DeLong’s test, and the final model was internally validated by bootstrap resampling. Results: Forty-one patients (28.5%) achieved discharge independence. Higher admission neutrophil-to-lymphocyte ratio (NLR) was independently associated with lower odds of discharge independence (adjusted odds ratio 0.47 per interquartile range increase, 95% CI 0.24–0.90; p = 0.022). Adding NLR to the clinical reference model improved discrimination (AUC 0.790 vs. 0.737; p = 0.039), with an optimism-corrected AUC of 0.767 after bootstrap validation. Other inflammatory indices did not significantly improve discrimination. Conclusions: In this single-center retrospective cohort of endovascularly treated patients with aSAH, admission NLR was independently associated with discharge independence and provided modest incremental prognostic information beyond established neurological severity scales. Full article

Glucose is the principal energy substrate for the brain. Hypo- and hyperglycemic episodes frequently occur in senescent people, contributing to functional and structural impairment of brain neurons and causing cognitive deficits in this population. In this study, we investigate whether long-term changes in the extracellular concentration of glucose affect viability and transmitter functions of septum-derived SN56 cholinergic neuronal cells through alterations in acetyl-CoA availability. Cells with low cholinergic expression (NCs) and cAMP/retinoic acid-induced high cholinergic expression (DCs) were investigated. Hypoglycemia brought about similar (approximately 20–30%) decreases in pyruvate dehydrogenase complex (PDHC) and ATP-citrate lyase (ACLY) activities and a 65% decline in lactate dehydrogenase (LDH) activity in NCs and DCs. Choline acetyltransferase (ChAT) and LDH activities in DCs were about 3–8 and 1.7–2.4 times higher than in NCs over the tested glucose concentration range, respectively. DCs appeared to be more resistant than NCs to hypoglycemia, as evidenced by lower glucose IC₅₀ values for cell count and intracellular LDH activity. On the other hand, some of functional properties of DCs, such as the cholinergic phenotype and their plasma membrane functions (trypan blue exclusion, TB+), were found to be more sensitive to hypoglycemia than those of NCs, as demonstrated by the higher IC₅₀ for glucose in DCs. Acetyl-CoA levels in DCs were 40% lower than in NCs, and decreased by about 25% with increasing hypoglycemia in both cell types. The cytotoxic effects of amyloid-β_25–35 (Aβ) and sodium nitroprusside (SNP; NO generator) were also tested. In 25 mM glucose medium, these toxic compounds exerted greater detrimental effects on DCs than on NCs. In contrast, in 1 mM glucose, more evident cytotoxicity of SNP and Aβ was observed in NCs. These data suggest that the higher rate of glycolysis in differentiated cholinergic septal neurons may be a protective mechanism against hypoglycemia. Full article

Background/Objective: With rising per capita sugar consumption, skin glycation-related issues including dullness, homeostasis disruption and accelerated wrinkling have gained widespread attention. However, globally standardized and rigorous evaluation criteria for anti-glycation efficacy remain lacking. This study aimed to establish stage-specific glycation injury cell models and elucidate the stage-dependent molecular mechanisms of glycation-induced fibroblast damage, providing a standardized reference for anti-glycation efficacy assessment. Methods: Three glycation injury models were constructed in human foreskin fibroblasts (HFF-1): early-stage (glucose-induced), intermediate-stage (glyoxal-induced), and late-stage (advanced glycation end products (AGEs)-induced). Core biomarkers including Nε-(carboxymethyl)lysine (CML), collagen type I (Col I) and elastin (ELN) were used to optimize modeling conditions via Cell Counting Kit-8 (CCK-8) and enzyme-linked immunosorbent assay (ELISA). Untargeted metabolomics based on ultra-high-performance liquid chromatography (UHPLC)-Q Exactive Orbitrap was applied to identify differential metabolites and perturbed pathways, following Metabolomics Standards Initiative (MSI) Level 2 identification criteria. Results: Optimal conditions were determined as 50 mmol/L glucose for 48 h, 0.5 mmol/L glyoxal for 48 h, and 200 μg/mL AGEs for 24 h. A total of 319, 34 and 148 differential metabolites were identified in the three groups, respectively. Six key pathways were significantly perturbed. Early and intermediate models shared similar mechanisms (purine metabolism disturbance), while the late model showed distinct alterations in pyrimidine, nicotinate, arachidonic acid and steroid hormone metabolism. Conclusions: Three stable stage-specific glycation models were successfully established in HFF-1 cells. Significant differences in metabolic profiles and mechanisms exist across the three stages, providing a rational basis for model selection and theoretical support for anti-glycation efficacy evaluation. Full article

Polycythemia vera (PV) is a JAK2V617F-driven myeloproliferative neoplasm characterized by erythroid expansion, increased thrombotic risk, and heterogeneous clinical outcomes. Although prior studies have described key transcriptional abnormalities—including Janus kinase–signal transducer and activator of transcription (JAK–STAT) hyperactivation and chronic myeloinflammation—most have examined single hematopoietic compartments. A multi-compartment approach may reveal conserved and lineage-specific disease-associated transcriptional programs. Here, an integrated, multi-compartment transcriptomic analysis of publicly available microarray datasets was performed, spanning bone marrow (BM) CD34+ progenitors, peripheral blood (PB) CD34+ progenitors, and whole blood from PV patients and healthy controls, with independent validation in neutrophils. Differential gene expression, pathway enrichment, and protein–protein interaction network analyses were used to delineate conserved versus compartment-specific transcriptional programs and to evaluate persistence of progenitor-derived signatures into mature myeloid cells. Across compartments, PV demonstrated consistent enrichment of inflammatory, interferon, and JAK–STAT-associated pathways despite limited overlap at the individual gene level, indicating that core disease processes are maintained through lineage- and differentiation-stage-specific transcriptional reprogramming. Network analysis identified highly connected hub genes, which were used to derive a single-sample gene set enrichment (ssGSEA) signature. This signature showed strong diagnostic performance across cohorts; remained enriched in PV neutrophils; and correlated with platelet count, indolent disease status, and reduced levels in post-splenectomy patients. Together, these findings support a model in which PV is driven by stable, progenitor-derived inflammatory programs that persist across myeloid differentiation while incorporating compartment-specific adaptations, and highlight the value of multi-compartment, network-based approaches for translational biomarker development. Full article

Post-infectious bronchiolitis obliterans (PiBO) is a chronic lung disease that develops after severe lower respiratory infections and leads to persistent inflammation and fibrotic changes in the small airways. In the present study, gene expression analysis was used to identify differentially expressed genes (DEGs) in sputum cells derived from PiBO patients and compare them to healthy controls. Clinical history, lung function parameters, and induced sputum samples were collected from nine patients with PiBO and eight healthy controls. Multiplex immunohistochemistry (mIHC) as well as mRNA sequencing (MACE-Seq) were performed. Evaluation of the biological targets was done by KEGG pathway enrichment analysis. PiBO patients showed significantly reduced lung function parameters, an increased neutrophil count, and an altered macrophage profile in sputum. Transcriptome analysis revealed significant upregulation of the TNFα-dependent NFκB signalling pathway, as well as significant downregulation of the oxidative phosphorylation (OXPHOS). Linear regression analyses and mIHC indicated a shift in macrophage polarisation that may contribute to the dysregulated gene expression. Notably, expression of these DEGs significantly correlated with FEV₁ lung function. These findings indicate a central role of macrophages in the immunopathology of PiBO and contribute to our understanding of the molecular mechanisms involved in the disease process. Full article

Background: Rapid and accurate preoperative diagnosis of periprosthetic joint infection (PJI) in patients undergoing revision arthroplasty remains challenging. Intraoperative differentiation between PJI and aseptic failure is limited by the lack of rapid and reliable diagnostic tools. Flow cytometric bacterial cell count of synovial fluid may represent a promising adjunct in this setting. This study explores the feasibility of using flow cytometric bacterial count of synovial fluid in the detection of PJI in patients during revision arthroplasty. Methods: In this exploratory pilot study, 93 patients provided informed consent. After exclusion of dry taps (n = 33), synovial fluid samples were collected intraoperatively from 60 patients undergoing revision total hip (THA) or total knee arthroplasty (TKA) at three hospitals in the Netherlands. Sixty samples were analyzed using the Sysmex UF-4000 to quantify flow cytometrically bacterial cells. Infection status was retrospectively determined according to the criteria of the European Bone and Joint Infection Society (EBJIS). Group differences were assessed using non-parametric analyses. Results: After exclusions due to dry taps and technical issues, samples of 43 patients could be analyzed. Of these patients, 16 had a confirmed PJI. Bacterial counts were significantly higher in patients with PJI compared with aseptic failure (median 1029 [IQR 287–4089] vs. 251 [IQR 92–433], p < 0.01). Conclusions: Flow cytometric synovial fluid analysis using the Sysmex UF-4000 demonstrated the ability to differentiate between patients with a PJI and with aseptic failure based on bacterial counts. However, this study was underpowered for diagnostic accuracy claims, and the feasibility of using flow cytometric bacterial count over established parameters such as total leukocyte and PMN% remains unclear at this stage. Full article

Meningiomas are the most common intracranial tumors. DNA methylation analysis in benign and aggressive meningiomas showed decreased MIR21 methylation and overexpression of hsa-miR-21-5p in atypical and anaplastic tumors. Transcriptomic analysis of distinct WHO grade meningiomas showed multiple predicted hsa-miR-21-5p target genes as differentially expressed. They were mainly related to processes of intercellular and intracellular signaling. Intercellular communication in meningioma was investigated using the deposited scRNA-seq dataset and deconvolution of our RNA-seq data. We found WHO grade-related differences in the microenvironment including inverse correlation between the count of border-associated macrophages (BAM) and the level of hsa-miR-21-5p. Single-cell transcriptomics suggests the role of interleukin 6 in direct communication between tumor cells and BAMs. IL6R and IL6ST are predicted targets of hsa-miR-21-5p downregulated in atypical/anaplastic meningiomas. IL6R downregulation was also confirmed by immunohistochemistry. Hsa-miR-21-5p enhanced proliferation and viability of KT21-MG1 meningioma cells and showed a regulatory effect on IL6R, IL6ST and other predicted target genes TIMP3, PIK3R, RHOB, and SASH1 by interacting with 3′UTRs. DNA hypomethylation-related overexpression of hsa-miR-21-5p contributes to aggressive meningioma growth by interaction with multiple target genes, and probably affects microenvironment communication between meningioma cells and BAMs by lowering the IL6R level in tumor tissue. Full article