Search Results (15,849)

The study was conducted to establish the thermoneutral zone (TNZ) of Murrah buffalo calves under precisely regulated environmental conditions. Six healthy calves (8–10 months) were exposed to progressively decreasing (24–15 °C; Temperature-Humidity Index (THI): 72.45–58.98) and increasing (25–40 °C; THI: 73.49–90.32) ambient temperatures with 3 °C intervals in a psychrometric chamber. Each exposure was maintained for 10 consecutive days under cyclic conditions (12 h exposure and 12 h thermoneutral recovery). Physiological responses and body surface temperature (BST) were recorded, and blood samples were collected on day 10 at 1500 h. Segmented regression analysis was performed using the SegReg (Oosterbaan, 2017) software program to identify lower and upper critical temperatures (LCT, UCT) and corresponding THI thresholds. LCTs ranged from 18.06 to 19.14 °C across different responsive parameters, with breakpoints (BPs) at 18.60 °C (SE_BP = 0.634) for pulse rate (PR), 18.15 °C (SE_BP = 0.945) for haemoglobin, 18.15 °C (SE_BP = 0.189) for lymphocytes, 18.24 °C (SE_BP = 0.404) for granulocytes, 19.14 °C (SE_BP = 1.09) for reactive oxygen species (ROS), 18.15 °C (SE_BP = 23.9) for superoxide dismutase (SOD), and 18.06 °C (SE_BP = 4.2) for cortisol; no distinct LCT was detected for respiratory rate (RR), rectal temperature (RT), total leukocyte count (TLC), or heat shock protein (HSP) 70/HSP90 expression. BST at different locations exhibited an LCT between ~20 and 21 °C. UCTs for PR, RR and RT were observed at 28.9 °C (SE_BP = 0.892), 28.6 °C (SE_BP = 0.843) and 29.2 °C (SE_BP = 1.16), respectively. Neither BST nor erythrocytic parameters exhibited a distinct UCT; however, BST increased progressively with increasing exposure temperature. TLC, lymphocyte and granulocyte % showed UCTs of 28.15 °C (SE_BP = 0.753), 28.15 °C (SE_BP = 0.497) and 32.65 °C (SE_BP = 14.20), respectively. The UCT for cortisol was observed at 28.15 °C (SE_BP = 5.52), whereas ROS and SOD showed UCTs of 28.75 °C (SE_BP = 4.67) and 34.15 °C (SE_BP = 6.55), respectively. HSP70 and HSP90 exhibited breakpoints at 35.35 °C (SE_BP = 1.32) and 34.30 °C (SE_BP = 2.11), respectively. Integration of key physiological and systemic responses (excluding BST) indicated a TNZ ranging from 19.14 °C to 28.15 °C. These findings provide a multi-level characterization of thermophysiological responses in buffalo calves and support the use of integrated biological indicators for defining thermoneutral conditions under controlled environments. The estimated TNZ of 19.14–28.15 °C provides a practical reference for managing Murrah buffalo calves. Producers should monitor housing temperature and consider ventilation, shade and air movement when temperatures approach or exceed ~28 °C while providing protection from cold when temperatures fall below ~19 °C. This may help reduce thermal strain and maintain normal physiological function. Full article

The detection of circulating tumor DNA (ctDNA) and circulating tumor cells (CTCs) is a minimally invasive approach for diagnosing and monitoring cancer. These liquid biopsy-based strategies enable the identification of primary or metastatic tumors and provide valuable information on tumor biology and treatment response. A variety of techniques are employed to analyze components obtained through liquid biopsy. While CTCs are typically detected using cell-based methods, ctDNA is commonly analyzed using highly sensitive molecular approaches, including quantitative PCR (qPCR), digital PCR (dPCR), and next-generation sequencing (NGS). Extracellular vesicles (EVs), which carry tumor-derived nucleic acids, proteins, and other biomolecules, are increasingly investigated as potential biomarkers, while tumor-educated platelets (TEPs) can reflect tumor-associated molecular changes and may provide additional information for cancer detection and monitoring. Furthermore, emerging multi-cancer early detection (MCED) approaches aim to identify molecular signatures associated with multiple cancer types from a single blood sample, highlighting the broader diagnostic potential of liquid biopsy. Due to the high specificity of tumors and somatic mutations, ctDNA serves as a real-time biomarker for tracking cancer progression. The advantages of ctDNA diagnostics include its low invasiveness and its capacity to detect cancer at early stages. In addition to confirming the presence of a tumor, liquid biopsy approaches facilitate the assessment of malignancy and the evaluation of treatment response. In the field of human medicine, ctDNA plays a pivotal role in diagnostic procedures. It is utilized not only for screening tests that detect the presence of cancer but also for the development of targeted treatment protocols for specific patients. These protocols are informed by the detection of genomic alterations and are designed to monitor the response to therapy over the course of treatment. Similarly, CTC, EV, TEP, and MCED approaches are being investigated as complementary tools for cancer detection, molecular characterization, prognosis, and longitudinal disease monitoring. In the domain of veterinary medicine, ctDNA has been instrumental in the diagnosis of various neoplasms, including osteosarcomas, hemangiosarcomas, lymphomas, canine mammary tumors, and melanomas. Other liquid biopsy components, including CTCs and EVs, also show promise for the detection and characterization of tumors in companion animals, although their clinical application remains less developed than in human medicine. Another significant application is in the detection of minimal residual disease (MRD), where a small number of cancer cells remain undetectable by conventional tests, such as blood counts. This underscores the significance of advanced ctDNA analysis. However, challenges persist, including the low concentration of ctDNA in blood, the low ratio of mutated to normal DNA fragments, potential contamination, biological variability, and the need for standardized protocols. This review synthesizes the current knowledge on liquid biopsy, including ctDNA, CTCs, EVs, TEPs, and emerging MCED approaches, and the potential for transferring these technologies from human medicine to animal medicine. Continued research is necessary to enhance the sensitivity and specificity of detection, which could facilitate early cancer diagnosis in animals and improve survival through timely treatment. Full article

Shannon entropy of single-cell transcript counts reflects composition and sampling. We tested whether interferon-beta (IFN-β) stimulation was associated with donor-consistent diversity changes after employing technical controls. We reanalyzed GSE96583 peripheral blood mononuclear cells from eight donors with lupus, treating donors—not the 8949 target cells obtained from them—as inferential units. Analyses included sampling without replacement at 562 unique molecular identifiers, a 750-molecule sensitivity analysis, Miller–Madow correction, IFN-gene removal, and exact compositional decomposition. Mean paired-donor raw-count plug-in entropy increased by 0.138 bits in CD14+ monocytes and 0.329 bits in FCGR3A+ monocytes and decreased by 0.080 bits in natural killer cells. Directions persisted across prespecified robustness variants. Monocyte increases accompanied higher evenness and lower transcript dominance; natural killer cells showed the opposite pattern, without consistent richness changes. Adding IFN activity to donor-, condition-, and technical-covariate-adjusted models increased R² by at most 0.0108. Entropy contributions strongly overlapped pseudobulk expression changes. In GSE194122, the implemented binary chromatin-accessibility entropy was determined by open-peak count, precluding biological cross-modality coupling claims. These findings support a lineage-divergent association within this single lupus cohort; independent replication is required before broader biological or diagnostic interpretation. Full article

Background: Accurate interpretation of neonatal complete blood count (CBC) parameters is vital for managing life-threatening conditions in newborns. However, clinical decisions in Eastern Sudan frequently rely on non-local Western-derived or manufacturer-provided reference intervals (RIs), thereby risking diagnostic errors. Therefore, this study aimed to establish the local neonatal hematological RIs for healthy, term newborns in Eastern Sudan. Methods: This hospital-based, cross-sectional study was conducted at New Halfa Hospital. Umbilical cord blood samples were collected from 250 healthy term singleton neonates born to mothers with uncomplicated pregnancies and deliveries. Samples were analyzed using a Sysmex KX-21 hematology analyzer (Sysmex, Kobe, Japan). In accordance with Clinical and Laboratory Standards Institute (CLSI) guidelines, sex-specific 95% RIs were initially estimated, followed by assessment of whether sex-specific partitioning was warranted by calculating the central 2.5th and 97.5th percentiles. Differences in the distributions of hematological parameters between males and females were evaluated using the Mann–Whitney U test. Results: Of the total 250 newborns (123 males, 127 females), no statistically significant differences were found between male and female newborns for any of the eleven evaluated hematological parameters, justifying the pooling of data to establish unified RIs. The established 95% pooled RIs and medians were as follows: white blood cell (WBC) count: 6.33–20.90 × 10³/µL (median: 11.15 × 10³/µL); red blood cell (RBC) count: 2.91–5.04 × 10⁶/µL (median: 4.02 × 10⁶/µL); hemoglobin: 11.20–16.52 g/dL (median: 13.90 g/dL); hematocrit: 36.30–55.30% (median: 43.70%); mean cell volume (MCV): 93.41–127.53 fL (median: 109.70 fL); platelets: 49.20–438.68 × 10³/µL (median: 251.00 × 10³/µL); mean platelet volume (MPV): 7.36–11.55 fL (median: 9.30 fL); platelet distribution width (PDW): 16.10–19.37% (median: 16.90%). Conclusions: The results of this study provide preliminary locally derived neonatal CBC RIs for Eastern Sudan. The absence of statistically significant sex differences in this sample supports consideration of pooled neonatal RIs. The lower reference boundaries for hemoglobin, leukocytes, and platelets identified here differ markedly from Western-derived values and those reported in high-altitude cohorts, underscoring the critical clinical need to use these population-specific intervals, which may reduce inappropriate classification arising from the use of non-local reference intervals. Full article

The peripheral blood transcriptomic profile associated with clinically observed cutaneous radiation injury (CRI) during fractionated radiotherapy remains incompletely characterized. We analyzed blood transcriptomes from 22 adults receiving radiotherapy: 10 with CRI and 12 without radiation-related skin injury at blood collection. RNA was assayed using two-color Agilent microarrays. Variance filtering retained 12,251 probes that were evaluated using four complementary analyses: preranked Gene-Set Enrichment Analysis (GSEA) for ranked gene-set enrichment, Gene-Set Variation Analysis (GSVA) for sample-level gene-set scoring, weighted gene co-expression network analysis for co-expression structure, and differential expression analysis at the probe level. GSEA identified positive enrichment of interferon-γ response and E2F targets and negative enrichment of heme metabolism. Consistently, GSVA showed higher interferon-response, E2F-target, unfolded-protein-response, and cholesterol-homeostasis scores and lower angiogenesis, xenobiotic-metabolism, protein-secretion, hypoxia, and heme-metabolism scores in participants with CRI. Ten modular co-expression networks met the data-derived CRI-correlation prioritization criterion. Positively correlated networks were enriched for negative regulation of response to wounding and complement/coagulation cascades. Intramodular connectivity and sample-level expression analyses prioritized KDM4C and PARPBP as exploratory high-connectivity candidates, with both retaining top-10 connectivity in 95.5% of leave-one-out iterations. Exploratory differential expression analysis identified 1842 probes, comprising 585 upregulated and 1257 downregulated probes in CRI. Genes mapped from downregulated probes were significantly enriched for ribosome, spliceosome, and oxidative phosphorylation pathways. Across complementary analytical frameworks, CRI was associated with a coherent peripheral blood profile characterized by interferon and cell-cycle enrichment; altered wound-response and complement/coagulation processes; reduced heme-metabolism, angiogenesis, and xenobiotic-metabolism scores; and downregulated transcripts involved in ribosomal function, RNA processing, and mitochondrial energy metabolism. Although the cross-sectional design and later sampling of CRI participants precluded full separation of CRI-associated differences from treatment-time and cumulative-exposure effects, these findings extend the molecular characterization of CRI and identify pathways and high-connectivity candidates for longitudinal assessment of reproducibility, temporal dynamics, and potential diagnostic or predictive relevance. Full article

Background: Practical localized resistance tasks involving functionally divergent muscle groups may produce different systemic biochemical responses, although neither task isolates a single muscle. Methods: In this exploratory, nonrandomized, order-balanced within-participant crossover study, 102 young adults completed bilateral seated plantar flexion and unilateral elbow flexion, with 51 participants assigned by alternation to each condition sequence. The analysis-ready biochemical dataset contained complete observations for 102 participants. Blood was collected before and 2–3 min after each condition. We analyzed plasma glucose and lactate and serum total cholesterol, HDL-C, LDL-C, triglycerides, creatine kinase, lactate dehydrogenase, GGT, and cortisol. Linear mixed-effects models included exercise condition, sampling time, study period, assigned sequence, and their respective interactions with sampling time as fixed effects. Participant and participant-specific study visit were included as random intercepts where supported by the model. The exercise condition × time interaction remained the primary contrast, and Holm correction was applied across the ten predefined biochemical outcomes. Linear mixed-effects models included exercise condition, sampling time, study period, assigned sequence, and their respective interactions with sampling time as fixed effects. Participant and participant-specific study visit were included as random intercepts where supported by the model. The exercise condition × time interaction remained the primary contrast, and Holm correction was applied across the ten predefined biochemical outcomes. Results: After adjustment for period and sequence and Holm correction across the ten primary interaction tests, cortisol remained the only statistically significant exercise condition × time interaction (149.219 nmol·L⁻¹, 95% CI 62.338 to 236.099; unadjusted p = 0.000821; Holm-adjusted p = 0.008). Glucose showed a negative interaction estimate (−7.013 mg·dL⁻¹, 95% CI −12.238 to −1.788; unadjusted p = 0.009), but did not remain significant after Holm correction (adjusted p = 0.078). No period × time or sequence × time effect remained statistically significant after correction. Sensitivity analyses were not fully concordant with the primary model, supporting cautious interpretation of the cortisol finding. Conclusions: These findings describe immediate responses to two non-equivalent multi-muscle tasks and cannot be attributed to selective soleus or biceps-brachii activation, fiber-type composition, or muscle-specific substrate use. Full article

Background/Objectives: Prenatal methamphetamine exposure may adversely affect fetal and neonatal outcomes, but its contribution to fetal and infant death remains incompletely understood. This study aimed to describe six fetal or infant deaths in which methamphetamine and amphetamine were detected and maternal methamphetamine exposure was considered a possible contributing factor. Methods: After obtaining the necessary approvals, medico-legal records, autopsy findings, and toxicological results were retrospectively reviewed for six cases from Bursa (n = 3) and Şanlıurfa (n = 3). Results: One case occurred in 2019, one in 2020, and four in 2021. Five cases involved preterm delivery between 24 and 32 gestational weeks, whereas Case 4 was born at 37 gestational weeks. Three cases were stillbirths, two infants died on postnatal day 4, and the term-born infant died on postnatal day 17. Methamphetamine and amphetamine were detected in postmortem specimens from all cases. Blood samples were available in five cases, with methamphetamine concentrations ranging from 19 to 256 ng/mL and amphetamine concentrations ranging from 4 to 42 ng/mL. In one case, only decomposition fluid was available. The organ damage attributable to methamphetamine and its role in the pathophysiology of death could not be fully established. Stimulant exposure was considered the most probable cause of death in Case 3, methamphetamine exposure was classified as the direct cause of death in Case 6, and methamphetamine exposure was considered a predisposing factor in the remaining four cases. Conclusions: Although methamphetamine and amphetamine were detected in all cases, these findings alone do not establish a causal relationship with death. Larger studies are needed to clarify prenatal and postnatal exposure pathways and to establish interpretive toxicological reference ranges for methamphetamine in fetal and infant populations. Full article

Background: Whole-blood transcriptomic profiling can capture systemic immune-response alterations associated with COVID-19 and may support host-response-based classification. However, evidence regarding the discriminatory value of targeted immune-gene panels remains limited, and in small, high-dimensional datasets, the timing of feature selection may substantially affect model performance and interpretation. Aim: This study aimed to evaluate whether NanoString Human Immunology Panel profiles could distinguish COVID-19 from healthy-control measurements and to compare full-dataset feature selection (FDFS) with leave-one-out cross-validation (LOOCV)-embedded feature selection (LEFS). Methods: Publicly available E-MTAB-8871 data comprising 579 genes and 32 whole-blood transcriptomic profiles were analyzed. The dataset included 22 longitudinal COVID-19 measurements obtained from three participants and 10 measurements obtained from 10 healthy controls. Elastic Net regularization was used for feature selection. Random Forest, XGBoost, and LightGBM classifiers were evaluated using sample-level LOOCV. Model performance was assessed using threshold-dependent, discrimination, and probability-based metrics. A separate exploratory LightGBM model was analyzed using SHapley Additive exPlanations (SHAP) to characterize feature contributions. Results: FDFS identified a fixed 40-gene set, whereas LEFS selected a mean of 42 genes per fold (range: 40–47). LightGBM correctly classified all 32 measurement-level profiles (derived from 13 unique participants: 10 healthy controls and three longitudinally sampled COVID-19 participants) in both frameworks, achieving area under the receiver operating characteristic curve (ROC-AUC) and area under the precision–recall curve (PR-AUC) values of 1.000 and Brier scores of 0.005 and 0.006 in the FDFS and LEFS frameworks, respectively. Random Forest achieved accuracies of 0.969 and 1.000, whereas XGBoost achieved an accuracy of 0.969 in both frameworks. SHAP analyses consistently identified AICDA as the dominant contributor to model predictions, followed by ARHGDIB. Conclusions: This exploratory analysis showed that targeted NanoString immune-response profiles contained a compact transcriptomic signal capable of distinguishing COVID-19 from healthy-control measurements within the analyzed dataset. These findings provide proof-of-concept evidence of internal measurement-level discrimination. However, because the COVID-19 profiles consisted of repeated measurements from only three participants, sample-level LOOCV did not constitute independent participant-level validation. External validation in larger cohorts comprising independently sampled participants is required. Given that the COVID-19 arm comprised only three independent participants, these biological findings should be regarded as hypothesis-generating and require validation in substantially larger independent cohorts. Full article

The diagnosis of Bluetongue Virus (BTV) in Argentina is primarily conducted via RT-qPCR and faces challenges due to the limited number of diagnostic laboratories, many of which are located in regions where BTV is exotic and far from primary sample collection areas. As a result, blood samples must be transported over considerable distances for analysis. This poses significant logistical challenges due to the specific transport conditions that are required. Consequently, the development of methodologies that optimize sample collection, preservation, and safe transport is of vital importance. The objective of this study was to standardize a sampling protocol for the detection of BTV using filter paper cards. This protocol was developed to ensure adequate preservation and transport of blood samples for diagnosis. Hydration with TE at 37 °C resulted in the lowest Cq among the tested treatments, with ~10 TCID₅₀/mL representing the lowest viral concentration that was consistently detected under the experimental conditions. The stability of the sample on the cards was evaluated through a series of storage trials at temperatures of 25 °C, 4 °C, −20 °C, and −80 °C for 1, 7, 30, 60, 120, and 360 days. Stability was verified up to 120 days at all temperatures. Furthermore, the absence of infectious material in the card eluate was confirmed. A preliminary field evaluation was conducted, in which blood samples from cattle and sheep were collected in tubes and on cards. According to the Cohen’s Kappa index, the agreement between both methods was nearly perfect. The standardized methodology signifies a substantial advancement in BTV diagnosis, as it facilitates the transportation of samples without the necessity of refrigeration and with adequate biosafety conditions without losing diagnostic capacity. Full article

Microbacterium species, a genus of aerobic Gram-positive coryneform rods, are rare causes of human infection with only a few sporadic cases reported worldwide. MALDI-TOF MS, 16S rRNA gene sequencing, and even whole-genome sequencing are used to identify the species in a clinical laboratory setting. The study reports a case of Microbacterium paraoxydans bloodstream infection and reviews the literature, including eight additional reported cases of M. paraoxydans infection. We summarize the characteristics of each case, highlight the identification methods, and identify recurrent characteristics for M. paraoxydans infection. A 65-year-old man presented with stable condition of esophageal cancer and long-term prednisolone therapy for psoriasis. A central venous catheter (CVC) was placed for years for treatment of esophageal cancer. The patient presented with fever and chills, and Microbacterium paraoxydans was identified by MALDI-TOF MS from blood culture samples via chemo port. After the chemo port was removed, vancomycin followed by levofloxacin was administered for the entire 14-day treatment course according to susceptibility tests. This study highlights that a long-term central venous catheter and immunocompromised status were common characteristics among the reported cases of Microbacterium spp. infection. The susceptibility of Microbacterium spp. is not fully characterized and varies across reports; gentamicin and trimethoprim/sulfamethoxazole appeared more consistently active, whereas susceptibility to vancomycin was variable. We report this case to highlight the increasing number of infections caused by Microbacterium spp. in the current era and to provide information on the treatment of M. paraoxydans. Full article

Hypertension (HTN) is often asymptomatic and difficult to detect using blood pressure (BP) measurements unless BP is substantially elevated. Given its association with metabolic alterations and complications, this study aimed to establish a metabolite risk score (MRS) as a molecular tool to complement BP-based diagnosis. Plasma samples and clinical data from healthy individuals and HTN patients were obtained through the Korea Biobank Network, and non-targeted metabolomics was performed. Eight HTN-associated key metabolites were selected by least absolute shrinkage and selection operator (LASSO) regression. An MRS was calculated as their weighted sum in the discovery set and subsequently validated in the replication set. The MRS showed strong discriminative performance for HTN status in the replication set [area under the curve (AUC) = 0.926, 95% confidence interval (CI): 0.876–0.976] and remained significantly associated with prevalent HTN after adjustment for age and BMI [odds ratio (OR) = 1.747, 95% CI: 1.317–2.318, p < 0.001]. At the MRS cut-off, classification accuracy in the replication set was approximately 84%, with 83.7% sensitivity and 84% specificity. The MRS also showed weak-to-moderate positive correlations with systolic BP in both the discovery set (r = 0.320, p = 0.001) and the replication set (r = 0.335, p < 0.001); however, these correlations were no longer statistically significant after adjustment for age and BMI. These findings support further evaluation of the MRS as a complementary molecular approach for HTN discrimination. Integration with other omics platforms may facilitate the development of more comprehensive molecular approaches for HTN. Further validation in larger prospective longitudinal cohorts is required before its potential clinical application. Full article

Background: Fulminant myocarditis (FM) is a life-threatening inflammatory cardiac disease in children, frequently triggered by viral infections. Although mechanical circulatory support has improved short-term survival, data on viral etiology and long-term outcomes remain limited, particularly in Asian populations. Methods: We conducted a mixed retrospective–prospective analysis of pediatric FM patients at the Children’s Hospital of Fudan University from 2015 to 2025. Clinical data from 2015 to 2023 were collected retrospectively, while data from 2024 onward were collected prospectively. Clinical data, microbiological findings (PCR and metagenomic next-generation sequencing), treatment strategies, and outcomes were extracted from electronic medical records. Follow-up data for survivors were collected through December 2025. Results: A total of 53 children with FM were included. Median age was 91.5 months; 43.4% were male, and 56.6% were female. Common presenting symptoms included fever (64.2%) and vomiting (56.6%). Microbiological evidence was identified in 11 patients (20.8%), with rhinovirus (5.7%) and influenza virus (5.7%) being the most frequent, followed by enterovirus (3.8%) in peripheral specimens. During hospitalization, 79.2% required mechanical ventilation, 64.2% received extracorporeal membrane oxygenation (ECMO), and 45.3% underwent continuous renal replacement therapy (CRRT). Intravenous immunoglobulin (IVIG) was administered to 84.9%. In-hospital mortality was 13.2% (7/53), and 17.0% (9/53) of patients were discharged against medical advice (DAMA). Among 35 followed patients (median 12.4 months), most achieved favorable cardiac recovery; however, persistent conduction abnormalities, structural cardiac changes, and neurological sequelae were observed in a minority. Conclusions: Pediatric FM carries substantial in-hospital morbidity and resource utilization, despite favorable recovery rates in most survivors. The low pathogen detection rate in peripheral blood and the lack of endomyocardial tissue sampling preclude definitive conclusions regarding the underlying etiology, whether active viral replication or immune-mediated injury predominates. Endomyocardial biopsy-based investigations are urgently needed to clarify the underlying pathobiology. Full article

Background: This study aimed to compare grazing behavior scores between adolescents with essential hypertension (HTN) and controls. It also investigated the associations of grazing behavior with obesity, emotion regulation difficulties (ERD), problematic smartphone use, and screen time. Methods: This cross-sectional case–control study enrolled 60 adolescents with essential HTN and 60 controls selected via frequency matching based on age and sex, all aged 12–18 years. The diagnosis of essential HTN was confirmed using 24-h ambulatory blood pressure monitoring (ABPM). Grazing behavior, ERD, problematic smartphone use, screen time and body mass index (BMI) were assessed in all participants. Group comparisons and associations between grazing behavior and the study variables were analyzed. Results: The essential HTN and control groups were comparable with respect to age and sex distribution (p > 0.05). The mean BMI in the essential HTN group was 31.18 ± 6.81 kg/m², which was significantly higher than that in the control group (23.35 ± 4.47 kg/m²) (p < 0.001). A significant positive correlation was found between total Grazing Questionnaire (GQ) scores and BMI (r = 0.252; p = 0.006), ERD (r = 0.187; p = 0.041), and problematic smartphone use (r = 0.315; p < 0.001). A statistically significant difference was observed between the essential HTN and control groups in total GQ scores [F(1, 117) = 4.85, p = 0.030, η²p = 0.040]. After adjustment for BMI, participants with hepatic steatosis had significantly higher total GQ scores (p = 0.019, η²p = 0.138), Grazing Behaviors subscale scores (p = 0.046, η²p = 0.107), and Uncontrollability subscale scores (p = 0.025, η²p = 0.123) than those without hepatic steatosis. Conclusions: Adolescents with essential HTN had higher grazing behavior scores than controls after adjustment for BMI. Grazing behavior was also associated with ERD and problematic smartphone use across the study sample. The findings regarding grazing behavior and hepatic steatosis warrant cautious interpretation, as the subgroup analysis was exploratory and included a relatively small number of participants. Full article

In post-mortem genetic identification, teeth represent one of the most reliable sources of nuclear DNA when other biological samples are unavailable, owing to their ability to protect genetic material from autolysis, microbial degradation, and environmental insults. The present study describes a forensic veterinary case report involving a Staffordshire Bull Terrier litter in which the appearance of blue-coated offspring from a phenotypically black sire and a blue dam prompted parentage verification. The sire died before formal ante-mortem sampling could be carried out. In accordance with Regulation (EC) No. 1069/2009, Article 15, the carcass was interred in the owner’s garden. Before burial, two teeth were collected as the sole biological material available for identification by the official veterinarian, ensuring an unambiguous chain of custody from death certification to laboratory analysis. DNA extraction was performed using two independent protocols: Method A, a silica-membrane-based purification (QIAamp DNA Kit, Qiagen), applied to dental pulp, and Method B, a decalcification-based workflow (T-Bone Ex Kit) followed by DNA extraction using EZ2 Connect Instruments (Qiagen). Quantification of the DNA samples obtained were done by fluorometry. Short Tandem Repeat (STR) genotyping was performed using the Canine Genotypes™ Panel 1.1 by Applied Biosystems (Thermo Fisher Scientific, Waltham, MA, USA) targeting the 18 autosomal ISAG-recommended microsatellite loci. Parentage assignments were carried out against reference STR profiles obtained from peripheral blood of the dam and offspring. The two methods, considering a different biological matrix and different operating conditions, allowed us to obtain the STR profile, but Method B showed quality electropherograms, improving the confidence and interpretation of all the markers analyzed. The differences observed between the two approaches may reflect a combination of factors, including the different dental matrices analysed, the quantity and quality of the starting material, tooth identity and tissue preservation, and differences inherent to the extraction procedures themselves; therefore, in this case, the more complete and clearer STR profile obtained with Method B should be considered a case-specific observation rather than evidence of a direct effect of the extraction method. Overall, regardless of the DNA extraction method used, the results did not exclude the biological paternity of the blue-mantled offspring. Furthermore, this case report provides practical methodological guidance for DNA recovery from canine teeth and supports their use as a reliable source of post-mortem DNA for STR-based parentage testing. Full article

Background/Objectives: Prompt identification of bloodborne pathogens and their antimicrobial resistance is critical for effective sepsis management. Direct molecular testing of blood offers faster results than blood culture; however, its clinical utility has been limited by suboptimal sensitivity and procedural complexity. This study evaluated an automated rapid multiplex PCR assay (BPID^® system) and compared its performance with gold-standard methods. Methods: This was a preliminary, single-centre diagnostic-agreement study. Forty-three whole-blood samples were collected from patients with suspected bloodstream infection at Tri-Service General Hospital, Taipei, Taiwan, between January 2025 and January 2026, and were analysed. Three molecular methods and two culture-based methods were compared: the BPID^® system, a Qiagen^®-based workflow, the BioFire^® FilmArray^® BCID2 panel, blood culture, and MALDI-TOF MS identification. Positive percent agreement (PPA) was calculated at the organism level against a composite reference standard. Analytical sensitivity (limit of detection, LoD) was assessed in whole blood spiked with serial dilutions of ten reference organisms. Results: The composite reference standard yielded 47 reference-positive organisms across 42 samples. The BPID^® system achieved a PPA of 95.74% (95% CI, 85.8–98.8), numerically similar to the post-culture BCID2 panel (93.62%; p = 1.00, exact McNemar test), and higher than the Qiagen^® workflow (72.34%; p = 0.001), blood culture (72.34%; p = 0.007), and MALDI-TOF MS (74.47%; p = 0.013). In a sensitivity analysis using a composite reference standard from which the index test was excluded, 46 organisms remained reference-positive and the PPA of the BPID^® system was 95.65% (44/46; 95% CI, 85.5–98.8), again numerically similar to the BCID2 panel (93.48%; p = 1.00). For antimicrobial resistance determinants covered by both panels, the BPID^® system reproduced the BCID2 determinant profile in all 16 specimens in which such a determinant was reported (100%), versus 10 of 16 specimens (62.5%) for the Qiagen^® method. Confirmed LoD values for the BPID^® system were 0.67–20.81 CFU/mL, an 8.8- to 24.3-fold improvement over the Qiagen^® method (6.51–224.61 CFU/mL). Conclusions: The automated BPID^® system substantially improves PCR-based diagnostic sensitivity for bacteremia and, by operating directly on whole blood, removes the culture-incubation step that post-culture molecular panels require. Because the study was not designed as an equivalence or non-inferiority trial and evaluated only 43 specimens, the similar agreement observed for the BPID^® system and the BCID2 panel should not be interpreted as demonstrated equivalence; these findings should be regarded as a preliminary clinical evaluation requiring confirmation in larger multicentre studies that also enrol culture-negative and uninfected controls. Full article