Search Results (6,231)

Background/Objectives: Urine samples are the most frequently analyzed specimens in clinical microbiology laboratories. Although urine culture remains the gold standard for diagnosing urinary tract infections, it is time-consuming and resource-intensive. Therefore, reliable screening methods capable of predicting urine culture positivity are needed to optimize laboratory workflow. Automated urine analysis based on flow cytometry enables efficient screening and identification of samples with a low probability of bacterial infection, thereby rationalizing microbiological testing. This study evaluated the usefulness of a multivariable approach to support interpretation of flow cytometry results following the implementation of the Sysmex UF-4000 urine flow cytometer. Methods: Routinely collected urine samples from outpatients and hospitalized patients were analyzed using the UF-4000 flow cytometer, with a positivity threshold of ≥100 leukocytes/µL. Urinary parameters were compared between samples with positive and negative cultures. Multivariable logistic regression was applied to identify independent predictors of a positive urine culture. Urinary sediment parameters, including leukocyte, bacterial, fungal, and squamous epithelial cell counts, were assessed as covariates. Results: Urine samples with positive cultures showed significantly higher leukocyte counts (median 355.0, IQR 146.5–1429.4) and bacterial counts (median 9805.2, IQR 1134.3–45,011.5). Fungal and squamous epithelial cell counts differed only slightly between groups, although the differences were statistically significant (p < 0.001). Leukocyte counts were higher in urine samples from which Gram-negative bacteria were isolated compared with samples containing Gram-positive bacterial isolates (p < 0.001). The multivariable model demonstrated the most favorable overall performance, combining high sensitivity with improved specificity and the highest negative predictive value (AUC = 0.927). Optimal cut-off values were 70 leukocytes/µL and 105 bacteria/µL. Conclusions: Leukocyte and bacterial counts were the strongest predictors of positive urine culture results. A multivariable model including only these two parameters demonstrated high diagnostic accuracy and may serve as a practical screening tool to identify urine samples with a low probability of bacterial infection. The implementation of this approach could support more efficient use of urine cultures and help optimize laboratory workflow. Full article

Background: Autologous fat grafting is widely used in reconstructive, aesthetic and regenerative surgery and often requires repeated applications. Cryopreservation of lipoaspirate enables autologous fat banking and off-the-shelf availability; however, its clinical implementation is limited by freezing-induced tissue injury, regulatory requirements and uncertainties regarding the optimal preservation protocol. Glycerol is a biocompatible cryoprotective agent with promising preliminary data. Nevertheless, the optimal concentration for lipoaspirate cryopreservation remains unknown. The aim of this study was to determine the optimal glycerol concentration for preservation of adipose tissue processed according to the Cell-Enriched Lipotransfer (CELT) protocol in clinically relevant volumes under GMP-compatible conditions. Methods: Lipoaspirates from 10 patients were processed by centrifugation according to the CELT protocol and allocated into experimental groups: fresh unfrozen control, frozen samples without cryoprotectant, frozen samples with PBS, and frozen samples supplemented with glycerol in concentrations ranging from 10% to 60%. Samples were cryopreserved using a controlled freezing rate at a temperature of −80 °C for 24 h. Large-volume cryopreservation was additionally performed with the best concentration of glycerol. Post-thaw tissue quality was assessed by resazurin assay of whole tissue, stromal vascular fraction (SVF) cell live/dead counting, and resazurin assay after short-term cell culture. Results: Glycerol supplementation improved post-thaw tissue viability compared with cryopreservation without cryoprotectant or with PBS alone. An optimal concentration range between 10% and 30% glycerol was identified, with highest preservation of metabolic activity and surviving cell yield observed at 20%. Higher glycerol concentrations resulted in a marked decline in tissue quality. Cryopreservation in large volume was feasible and did not impair post-thaw viability compared with small-volume samples. Conclusions: Glycerol-based cryopreservation allows effective and GMP-compatible preservation of human lipoaspirate. An optimal glycerol concentration range was identified, enabling large-volume fat banking without compromising tissue quality. This protocol provides a clinically applicable strategy for autologous fat storage and may facilitate repeated reconstructive and regenerative treatments. Full article

The objective of the present study is to evaluate the effects of lactation stage, age, somatic cell count (SCC), and daily milk yield on plasmin–plasminogen (PL–PG) system activity and physicochemical milk traits in intensively reared Chios and Frizarta ewes. A total of 52 purebred ewes (26 ewes per breed and farm) were randomly selected and prospectively monitored during the 3rd, 5th, and 6th month post-lambing. Daily milk yield and body condition score (BCS) were recorded, and individual milk samples were collected for the assessment of PL–PG activities using enzymatic assays, SCC, electrical conductivity (EC), refractive index (RI), and pH. Correlation analysis and mixed linear regression models were used for the assessment of the effects. Lactation stage significantly affected PL–PG system traits in both breeds, but in opposite direction; plasmin and plasminogen plus plasmin declined toward late lactation in Chios ewes, whereas it increased in Frizarta ewes. Lower SCC was associated with reduced plasmin system activity in Chios ewes, whereas no effect was observed in Frizarta ewes. The plasminogen-to-plasmin ratio remained stable across lactation, breeds, and SCC classes, indicating coordinated regulation of the system. BCS was positively associated with plasmin activity during late lactation, suggesting a stage-dependent metabolic modulation. EC and pH were closely associated with SCC, while RI mainly reflected compositional variation. Our findings underline that, although the PL–PG system is primarily affected by lactation stage and mammary health status in sheep, there are breed-specific regulatory patterns which should be further investigated. Full article

Background: An unfavorable course of SARS-CoV-2 infection can lead to significant morbidity and mortality. The study aimed to develop a simple, accessible, and reliable tool to anticipate the poor results among COVID-19 pneumonia patients. Methods: This retrospective cohort study involves 306 individuals with severe COVID-19 pneumonia enrolled between March 2021 and June 2021. Each patient had confirmed SARS-CoV-2 infection and required oxygen therapy. Differential blood count and serum CRP were taken on admission day. Medical data were collected from the hospital’s information system. Results: Of 306 patients (133 females, 173 males, aged 66.3 ± 15.2 years), 105 (34.3%) died. Counts of neutrophils, lymphocytes, and eosinophils differed significantly between survivors and deceased (p < 0.001; p = 0.002; p = 0.009, respectively) and had substantially differentiating properties in ROC analysis. Built with the counts of neutrophils, lymphocytes, and eosinophils, the White Blood Cell Score (WBCS) was developed. WBCS robustly predicted mortality (OR = 2.821; CI: 2.037–3.906; p < 0.001) in the investigated population. Cumulative risk of death according to WBCS (ranging from 0 to 3 points) was as follows: 0 points—10.9%, 1 point—23.5%, 2 points—33.1%, 3 points—34.1%. Conclusions: Based on differential blood count, the proposed WBCS is easy to use and can be helpful in predicting mortality among severe COVID-19 patients. Full article

The bioactive compounds in thyme essential oil (TEO) have been investigated as natural preservatives. However, their direct application in foods is limited by their poor water solubility and high volatility. In this context, nanoemulsions represent promising delivery systems for bioactive compounds due to their improved physicochemical stability and functional performance. This study aimed to develop and characterize TEO nanoemulsions prepared by ultrasound-assisted encapsulation using an ultrasonic probe and whey protein concentrate as a surfactant, with potential application in chicken burgers. Different sonication times (1, 3, 5, 7, and 10 min) were evaluated, and ultrasonication time was evaluated as the experimental variable. The formulation processed for 3 min presented the smallest hydrodynamic diameter (289 nm) and a homogeneous spherical morphology. The nanoemulsions showed low cytotoxicity, maintaining cell viability above 90% at all evaluated concentrations. In vitro antibacterial assays demonstrated activity against Staphylococcus aureus and antifungal effects against Aspergillus and Penicillium species. When applied to chicken burgers, the treatment containing 100 ppm of nanoencapsulated TEO contributed to reductions in S. aureus and mesophilic aerobic microorganism counts during 7 days of refrigerated storage. These findings indicate that TEO nanoemulsions present potential as natural antimicrobial systems for food preservation applications. Full article

Autophagy-mitophagy pathways are essential for regulating immune homeostasis. However, their contribution to population-level chronic low-grade systemic inflammation (SI) remains unclear. The objective was to investigate the association between variation in the genes related to the autophagy-mitophagy pathways and SI, and to examine whether lifestyle factors modify this relationship. We conducted genome-wide association studies and gene-set enrichment analyses using data from the Korean Genome and Epidemiology Study (KoGES, n = 28,102) and UK Biobank (UKBB, n = 343,892). SI was defined as an elevated white blood cell count or high-sensitivity C-reactive protein. Using Core Longevity State Vectors (CLSVs)—gene sets representing immune-longevity pathways derived from comparative transcriptomic analysis—we tested six pathways and constructed a weighted genetic risk score (GRS) from significant variants. Gene–lifestyle interactions were examined with respect to major dietary and lifestyle factors. Among six CLSVs, only CLSV-2 (mitophagy and autophagy) showed a significant association with SI (β = 0.425, p = 0.008). Six single nucleotide polymorphisms (SNPs) in autophagy-mitophagy genes (INPP5D, ATG16L1, ATG7, AP3S1, OPTN, and VPS33A) were associated with SI in KoGES (p < 5 × 10⁻⁵), and ten SNPs (genes selected in KoGES plus RAB7A, ATG12, VPS33A, BECN1) reached genome-wide significance in UKBB (p < 5 × 10⁻⁸). A higher GRS was associated with increased SI in both cohorts and was strongly associated with metabolic syndrome (MetS, OR = 1.91 in KoGES; OR = 1.62 in UKBB). SI was characterized by neutrophilia with relative lymphopenia. In UKBB, significant gene–lifestyle interactions were observed for diet, physical activity, smoking, and alcohol (p < 0.01). Favorable lifestyle factors reduced SI most effectively in individuals with protective genotypes. Among individuals with a high vegetable/fruit intake, SI prevalence was 35%, 36%, and 38% in the negative-, zero-, and positive-GRS groups, respectively, compared with 36%, 45%, and 48% in the low-intake groups. In conclusion, genetic variations in autophagy-mitophagy pathways specifically influence SI. Genetic predisposition substantially modifies the benefits of lifestyle, underscoring the importance of integrating genetic and lifestyle factors in understanding SI susceptibility. Full article

(1) Background: The search for new polymodal antioxidants to correct oxidative stress of various origins and its consequences remains one of the most pressing and rapidly developing areas of biomedical research. (2) Methods: Hydrogen peroxide and hydroxyl radical detection, induced luminescence assay, ELISA for 8-oxoguanine detection, animal survival, blood cell count, micronucleus test, and PCR were used. (3) Results: 2R,3R-trans-dihydroquercetin (DHQ) was shown to reduce the amount of hydrogen peroxide and hydroxyl radicals formed during water radiolysis, leading to reduced damage to biomolecules. DHQ is a radioprotector, most effective at a dose of 300 mg/kg administered 15 min before radiation exposure. The dose reduction factor is 1.22. DHQ administration reduces the severity of radiation-induced leukopenia and thrombopenia by protecting red bone marrow cells. The mechanism of DHQ’s radioprotective action is fundamentally different from that of classical stress response inducers and is based on the normalization of the target cell transcriptional profile, rather than its hyperstimulation. (4) Conclusions: DHQ’s ability to restore the expression of antioxidant defense, DNA repair, and apoptotic genes to physiological levels under radiation exposure allows it to be considered a promising pharmacological agent for the correction of radiation-induced damage to normal tissues. Full article

Helicobacter pylori is a Gram-negative bacterium that inhabits the gastric mucosa, with a global prevalence in humans of approximately 40%. It is likely the cause of 90% of gastric cancer (GC) cases and thus considered the most prominent driver of GC development. However, during gastric mucosal atrophy, other bacteria such as nitrate-reducing bacteria (NRB) also proliferate. In this study, we isolated NRB from patients with gastritis and GC to examine their effects on the epithelial cell cycle and production of various cytokines in monocytic cell lines. Bacterial counts (excluding H. pylori and NRB) increased with the progression of gastric mucosal atrophy and were significantly higher in patients with GC. Gastric epithelial cell lines were stimulated with isolated NRB, and the proportion of cells in each cell cycle was measured. Strains from patients with open-type gastritis progressed more rapidly through cell cycles than those from patients with GC. NRB isolated from gastric cancer had high nitrate-reducing activity. Thus, NRB may contribute to GC progression during H. pylori-induced carcinogenesis. Therefore, evaluating gastric atrophy and microbiota may be important for managing the risk of GC. Full article

Hemotherapy in small ruminants is indicated for several acute and chronic conditions; however, its clinical use is often limited by the difficulty in identifying suitable donors, particularly regarding blood volume availability and hematologic compatibility. Xenotransfusion in small ruminants with bovine blood may represent a practical alternative in emergency situations involving severe anemia when homologous donors are unavailable. This study evaluated the clinical, hematologic, biochemical, and blood gas responses of sheep subjected to acute blood loss followed by bovine whole blood xenotransfusion. Six healthy adult castrated male sheep (mean body weight 44.3 ± 7.2 kg) underwent removal of 40% of their estimated total blood volume. Parameters were assessed before hemorrhage induction (T0) and at times T30, T6h, T12h, T24h, T48h, T72h, T96h, T5d, T6d, T7d, T8d and T16d after transfusion. Acute blood loss significantly reduced packed cell volume and erythrocyte count at T0 (p < 0.05). After xenotransfusion, packed cell volume increased at T30min, T6h, and T12h and remained stable until T72h (p < 0.05), with progressive erythrocyte recovery and sustained macrocytosis. Total leukocyte count remained unchanged, whereas platelets increased at T7D (p < 0.05). Total protein decreased at T0 and subsequently increased. Transient elevations in urea, creatinine, glucose, pO₂, and SO₂ were observed (p < 0.05), without acid–base imbalance. Clinical parameters progressively stabilized, and no severe transfusion reactions occurred. Bovine whole blood xenotransfusion may represent a promising therapeutic alternative for sheep subjected to acute blood loss under the experimental conditions evaluated in this study. The procedure was associated with improvements in clinical, hematological, and biochemical parameters, and no severe transfusion reactions were observed during the monitoring period. These findings support the potential clinical applicability of this approach as an emergency intervention in situations where homologous donors are not readily available. Full article

Sporulation represents a complex metabolic reprogramming process in bacteria. In this study, we used CRISPR-Cpf1 to knock out spoIIE and rsfA in Bacillus licheniformis. The ΔspoIIE strain completely lost sporulataion capacity, while ΔrsfA showed a 25% reduction. Although viable cell counts decreased by 80.7% and 45.7%, respectively, glucose consumption and 2,3-butanediol synthesis remained unchanged, and acetoin synthesis increased by 19% in ΔspoIIE. Per-cell metabolic rates were significantly enhanced: glucose uptake increased 2.7–3.4-fold, acetoin synthesis 2.3–4.2-fold, 2,3-butanediol synthesis 1.7-fold, and heterologous protein expression 10–15-fold. These findings demonstrate that blocking sporulation liberates metabolic resources and enhances the specific productivity of vegetative cells, providing a strategy for engineering high-performance B. licheniformis cell factories. Full article

Semen parameters, including sperm counts, have rapidly declined in men across the globe over the last five decades. Although this decline remains unexplained, lifestyle factors may affect male fertility. Recently, several studies highlighted a potential link between non-steroidal anti-inflammatory drug (NSAID) usage, such as naproxen and ibuprofen, and declining male fertility. However, the mechanisms by which these common analgesics affect male fertility, including their effects on the blood–testis barrier (BTB), remain poorly characterized. Utilizing an in vitro rhesus macaque non-human primate (NHP) BTB model, we demonstrate that serum levels of naproxen and ibuprofen alter the function of BTB. Following short-term naproxen and ibuprofen treatment of NHP primary Sertoli cells, we show that these NSAIDs increase the transepithelial electrical resistance, indicating an overall strengthening of the Sertoli cell junctions. Furthermore, naproxen and ibuprofen treatment alter the expression of genes involved in maintaining the BTB. Specifically, the genes that were significantly expressed in response to ibuprofen exposure were enriched for human phenotypic abnormalities linked to male factor infertility. Together, these results suggest that short-term naproxen and ibuprofen treatment disrupt the function of the BTB by altering the integrity of the Sertoli cell junctions, proposing a potential role of NSAIDs in male factor infertility. Full article

Infrared thermography has been proposed as a non-invasive tool for mastitis detection in dairy ruminants; however, the extent of environmental confounding and diagnostic performance in small ruminants remain poorly characterized. This study evaluated udder thermography in dairy goats through correlation analysis under winter and summer conditions, and an experimental intramammary inflammation challenge using Staphylococcus aureus lipoteichoic acid, with receiver operating characteristic analysis using somatic cell count >1500 × 10³ cells/mL as the reference standard. Strong positive correlations between ambient temperature and udder surface temperatures intensified substantially from winter to summer, while surface temperatures showed weak or absent correlations with rectal temperature. Experimental inflammation induced a 12-fold increase in somatic cell count (305 vs. 3658 × 10³ cells/mL, p < 0.001); however, thermographic responses remained minimal and spatially inconsistent, with area under the curve values approximating 0.5 and weak correlations between thermographic measurements and somatic cell count. Environmental temperature variation and physiological thermoregulatory adjustments substantially exceeded the minimal thermal signal generated by intramammary inflammation, limiting diagnostic utility. Infrared thermography showed poor diagnostic utility for detecting experimentally induced intramammary inflammation in dairy goats under the tested conditions. Full article

Background/Objectives: Corneal endothelial cells (CEnCs) exist on the inner surface of the cornea and regulate its hydration. The immune system cannot penetrate CEnCs easily because the cornea is avascular and anterior chamber-associated immune deviation suppresses the immune reaction. Nevertheless, inflammatory cells can infiltrate through the corneal stroma and anterior chambers, and corneal endothelial inflammation can occur. In this cross-sectional study, we investigated the association between the neutrophil-to-lymphocyte ratio (NLR) and human corneal endothelial cells (CEnCs). Methods: A total of 307 eyes from 307 subjects who underwent specular microscopy were included. Corneal endothelial cell density (CECD), hexagonality (HEX), central corneal thickness (CCT), coefficient of variation (CV), and cell area were measured preoperatively using specular microscopy. Whole blood samples were obtained to measure the complete blood cell count. The NLR was calculated, and its relationship with CEnCs was evaluated. Results: In all subjects, CV was positively correlated with the percentage (%) of neutrophils (r = 0.120, p = 0.037) and absolute neutrophil count (r = 0.131, p = 0.022) and negatively correlated with the % of lymphocytes (r = −0.131, p = 0.022). HEX was correlated with the % of neutrophils (r = −0.156, p = 0.006), % of lymphocytes (r = 0.141, p = 0.014), % of basophils (r = 0.142, p = 0.013), the NLR (r = −0.129, p = 0.024), and the mean corpuscular volume (r = 0.121, p = 0.035). Conclusions: CV and HEX, which indicate the stability of CEnCs, are associated with NLR in the peripheral blood, suggesting that systemic inflammation and immunity may implicate in the pathology of CEnCs. Full article

Hafnia alvei, which belongs to the Enterobacteriaceae family, has been occasionally documented in animal infections but is still not well characterized in the context of bovine mastitis. This research examined the prevalence and antimicrobial resistance characteristics of H. alvei in dairy cows suffering from subclinical mastitis in South Africa’s Free State Province. In the Thabo Mofutsanyana District, a total of 174 milk samples were obtained from cows on six different dairy farms. The California Mastitis Test (CMT) was used to screen for subclinical mastitis, and somatic cell count was used to confirm it. Standard culture methods were used for bacterial isolation, and presumptive Enterobacteriaceae isolates were identified through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Out of the 174 samples, 84 (48.2%) tested positive for CMT, and 68 (39.1%) met the SCC criteria for subclinical mastitis at a cow level, while 96/336 (28.5%) were infected at a quarter level. Of the 100 presumptive Enterobacteriaceae isolates, 33 (33.0%) were identified as H. alvei (p = 0.0034). Antimicrobial susceptibility testing showed that 50% of the isolates were resistant to penicillin, followed by tetracycline and erythromycin with 25% and 10%, respectively. Furthermore, the results showed that 17 (51.5%) isolates exhibited multidrug-resistant profiles. The results suggest that H. alvei could be a contaminant in raw milk associated with bovine subclinical mastitis in this area, necessitating additional epidemiological research that includes healthy matched controls. Full article

Tuft cells (TCs) are rare chemosensory epithelial cells that regulate mucosal homeostasis in multiple organs, but their role in salivary gland (SG) biology remains poorly defined. This study aimed to define TC structure in mice submandibular glands (SMGs), determine how TC loss affects gland organization and function, and evaluate whether TC abundance in human minor SGs is associated with Sjögren’s disease (SjD) features. Specifically, TC ultrastructure and ductal localization were characterized in female and male C57BL/6J mouse SMGs by transmission electron microscopy and immunostaining. Wild-type and C57BL/6J-Pou2f3^-/- (TC-deficient mouse strain) SMGs were analyzed by histology and bulk RNA-seq, and salivary function was assessed by saliva flow and proteomics. Human minor SG biopsies from SjD and non-Sjögren sicca (nSjD) patients were analyzed by immunostaining and Poisson regression. In mice SMGs, TCs showed conserved ultrastructural features and localization in both sexes. TC loss was associated with marked sex-biased transcriptome remodeling, morphological disruption, and altered saliva quantity and quality. In humans, TC counts differed between nSjD and SjD groups and were associated with salivary flow, serologic status, and histopathologic features. These findings support a role for TCs in SG epithelial integrity and suggest TC abundance as a candidate biomarker of SG dysfunction. Full article