Herd-level occurrence and spread of Theileria equi and its associated ticks on an Austrian horse farm.

Brain lithium temporospatial kinetics in bipolar disorder: a repeated-measures 7Li MRI study of dosing regimens in Germany.

Cumulative dose-based management of doxorubicin (DOX) does not necessarily reflect the true systemic exposure to DOX or its cardiotoxic metabolite, doxorubicinol (DOXol). The aim of this study was to develop and validate a regression-based limited-sampling strategy to estimate the areas under the concentration-time curves (AUCs) of DOX and DOXol in patients with diffuse large B-cell lymphoma receiving CHOP chemotherapy. Forty-five patients receiving CHOP therapy were enrolled in this study. Plasma samples were collected at 7 time points between 1.5 and 25.5 hours post-DOX administration. AUCs were calculated using noncompartmental analysis. Linear regression models were constructed using plasma concentrations from one or 2 time points. Predictive performance was assessed using 5-fold cross-validation and evaluated by mean error, mean absolute error, and root mean square error. All sampling time points showed significant correlations with the full AUCs for DOX and DOXol, with the strongest being observed at 13.5 hours. Two-point models incorporating 1.5 and 13.5 hours concentrations yielded the highest predictive accuracy (R2 = 0.968 for DOX and 0.993 for DOXol), with low %MAE and %RMSE values. These models enabled simultaneous, accurate AUC estimation for both analytes with minimal sampling. Regression-based limited-sampling strategy using one or 2 sampling time points is a feasible and accurate method for estimating the AUCs of DOX and DOXol in clinical settings. This strategy enables the practical assessment of cumulative exposure beyond conventional cumulative dose-based management and may support individualized dosing of DOX.

The current use of artificial light during the natural dark phase has acquired contaminant dimensions, known as "light pollution". It is well known that exposure to dim light at night (dLAN) during the postnatal period severely impairs the immune system and related organs, but few reports have demonstrated the effects of dLAN during the fetal period. This study, therefore, examines whether exposure to dim light at night during two critical developmental windows (i.e., prenatal and postnatal periods) leads to long-lasting dysregulation of circadian, behavioral, and immune organization, as well as spleen immune responses, in early adulthood. To address this question, these outcomes were assessed using two defined sampling time points. To answer this question, we exposed two groups of C57BL/6J male mice to dim night light during the gestational and postnatal periods and compared them with control groups that were exposed to light-dark conditions (12 h each, LD). Parametric and non-parametric activity/rest values were analyzed with circular statistics. Compared to their controls, we found differences in alpha, onset, offset, M10, and L5 start time in dLAN groups. We also assessed the transcript levels of clock genes and inflammatory mediators in spleen tissue and found a dampening of daily variation in mRNA expression in both experimental groups. Finally, we used an ovalbumin (OVA) allergy challenge to test the B-cell response in the spleen and found a significantly higher cell recruitment to the spleen and more anti-OVA IgE. Together, these results clearly show that dLAN, at two ZT sampling points, affects peripheral molecular clocks and responses in the spleen, and that these effects are independent of the life stage at which exposure to dim light at night occurs.

Recent evidence suggests that heat treatment (HT) and resistance training can limit skeletal muscle mass loss during immobilization. However, the effects of repeated HT sessions combined with hybrid exercises (EX), which promote both endurance and resistance responses, on muscle protein turnover following hindlimb unloading (HU) remain unexplored. This study investigated the effects of reloading strategies using HT and/or EX on protein synthesis, ribosome biogenesis and autophagic markers. Eight-week-old male C57BL/6J mice underwent HU (3weeks), HU+HT (1week, 5 sessions, 30min, 40°C), HU+EX (5 sessions of high-intensity inclined treadmill running), HU+EX+HT or HU+REL (passive reloading). HU+HT did not induce additional effects compared with the HU+REL group concerning protein synthesis rates, although one marker of protein synthesis (phosphorylated ribosomal protein S6) tended to increase. HU+HT did not lead to an increase in rRNA content. HU+EX induced a concomitant decrease in the mitochondrial fission markers phosphorylated dynamin-related protein 1 and phosphorylated AMP-activated protein kinase, suggesting improved mitochondrial efficiency. However, no additional effects were observed for rRNA or heat shock proteins compared with the HU+REL group. Finally, exercise followed by heat treatment post-HU (HU+EX+HT) appears to be detrimental, as protein synthesis rates did not increase, despite an increase in rRNA content. Thus, at this sampling time point (14days post-HU), neither heat therapy nor hybrid retraining produced additional effects on the studied markers compared with a normal resumption of activity. In contrast, the sequential application of the two stressors may inhibit post-HU muscle mass recovery, as evidenced by the absence of increased protein synthesis rates.

The c-Met signalling pathway is dysregulated in non-small cell lung cancer (NSCLC), with c-Met protein overexpression emerging as a potentially actionable biomarker given the development of c-Met-targeted antibody-drug conjugates. This retrospective study assessed the consistency of c-Met protein overexpression over time in patients with non-squamous NSCLC and two or more longitudinal biopsy samples (one before treatment, one post-treatment). Samples from 160 patients were analysed for c-Met protein expression. The concordance rate for c-Met protein overexpression between samples was 81.3%, indicating most patients had consistent expression status. Patients with an EGFR mutation had a lower concordance rate (65.8%). Fourteen patients with high discordance (negative to positive) between sampling time points showed a high frequency of oncogenic driver alterations, hence often received targeted therapy before the second sample and had high percentages of cells at 2+ intensity in c-Met immunohistochemistry before treatment. These results indicate c-Met protein overexpression can be assessed before or after treatment since most patients maintain consistent c-Met status. As targeted therapies may elevate c-Met overexpression over time, retesting may be necessary in those with an oncogenic driver alteration initially diagnosed as c-Met negative.

Catecholamine metabolism contributes to energy homeostasis and may differ in low-weight conditions of various origins. Free plasma metanephrines, 3-methoxytyramine (3-MT), normetanephrine (NM), and metanephrine (M), are stable end-products of dopamine, noradrenaline, and adrenaline metabolism, respectively, with potential circadian rhythmicity. However, their 24-hour profiles remain unknown in anorexia nervosa (AN) and constitutional thinness (CT). This exploratory study aimed to assess 24-hour variations of plasma free metanephrines in women with acute AN (n = 10), recovering AN (AN-Rec, n = 11), CT (n = 10), and healthy controls (n = 10), and to explore their association with biomarkers of undernutrition severity and weight regulation. Participants underwent 24-hour in-patient evaluation, including standardized meals and activity, and twelve time-point plasma sampling. Free metanephrines were measured by LC-MS/MS. 24-hour rhythmicity was analyzed using cosinor regression. AN and CT showed higher mean 24-hour levels for 3-MT, NM, and M than controls, with variable patterns depending on metabolites. AN-Rec displayed higher 3-MT levels than controls only. Cosinor analysis revealed a significant 24-hour variation consistent with a circadian pattern for 3-MT with an acrophase around 13:00 in AN, AN-Rec, and CT. NM showed a circadian pattern in AN, CT, and controls. Mean 3-MT, NM and M correlated negatively with BMI and IGF-1, and positively with ALT and vitamin B12. Midday elevations of free plasma metanephrines may reflect metabolic stress and hepatic alterations in AN. Their increase may may reflect distinct underlying physiological mechanisms in both AN and CT. Further studies are needed to confirm these exploratory findings.

Early identification of acute kidney injury (AKI), including subclinical AKI, remains a critical challenge in septic patients undergoing major non-cardiac surgery. Exosomal microRNAs (miRNAs) represent emerging biomarkers capable of reflecting early renal cellular stress before detectable changes in serum creatinine occur. This prospective observational cohort study enrolled 120 adult septic patients undergoing major non-cardiac surgery (> 2h) at Imam Sajjad Hospital, Tabriz, between early 2025 and August 2025 (Ethics Approval ID: IR.IAU.TABRIZ.REC.1404.179). Peripheral blood was obtained at three perioperative time points (preoperative, intraoperative, and 24h postoperative) for exosome isolation and miRNA quantification. Clinical AKI was defined according to KDIGO criteria. Subclinical AKI was defined as urinary TIMP-2 × IGFBP7 > 0.3 in the absence of any rise in serum creatinine. Diagnostic performance of candidate miRNAs was assessed using receiver operating characteristic (ROC) analysis based on observed patient data. Among the 120 enrolled patients, a proportion developed AKI during the 24-hour postoperative observation period (final numbers provided in the Results section). Patients who developed AKI demonstrated distinct perioperative alterations in specific exosomal miRNAs compared with those who did not develop AKI. Several miRNAs showed measurable discriminatory ability for early AKI detection across different sampling time points, with areas under the ROC curve (AUCs) ranging within clinically relevant thresholds (AUC values reported in the Results and Tables following recalculation using observed data with 95% confidence intervals). These findings suggest that perioperative plasma exosomal miRNAs may serve as promising early indicators of acute kidney injury in septic patients undergoing major surgery. Their potential role in identifying subclinical AKI warrants further investigation in studies specifically powered for this endpoint.

Temporal Sampling of Commercial Turkey Flocks Highlights Challenges of Preharvest Salmonella enterica Monitoring.

Coxiella burnetii shedding in dairy cattle and its correlation with humoral and cell-mediated immune responses.

This study investigates the dynamics of break loose glide force (BLGF) in prefilled syringes (PFS) and injection time in autoinjectors (AI) across different orientations and time points, focusing on their impact on drug delivery performance. PFS and Autoinjectors are pivotal in modern therapeutics, enabling precise administration of biologics, vaccines, and other sensitive pharmaceuticals. Patients with chronic conditions and individuals with limited dexterity requiring frequent self-administration, all benefit from devices that minimize BLGF to ensure consistent injection times. To explore the influence of storage orientation (tip-up vs. tip-down) and time on these devices, we conducted a comprehensive evaluation over a 12-month period supplemented by reference timepoint samples. Utilizing a dual-orientational framework, PFS and autoinjectors were stored under controlled conditions to assess orientation-dependent performance. Aged and empty samples served as reference controls. The study's primary metrics included BLGF, which is the initial force required to initiate plunger movement then enable injection along the barrel, and injection time. These parameters were analyzed using a calibrated Zwick/Roell mechanical tester under controlled environmental conditions.Key findings reveal statistically significant but not practically impactful effects of storage orientation on BLGF or injection time across all tested intervals. Minor time-based variations were noted, with BLGF shifts being less than 1 N and injection time deviations under 2 seconds. These changes indicated no practical significance or risk of out-of-specification performance. The Analysis of variance (ANOVA) analysis confirmed no evidence suggesting orientation affects functionality.This work demonstrates that short to mid-term storage conditions do not adversely affect device performance, providing critical assurance for manufacturers, regulators, and end-users. The study highlights the robustness of PFS and Autoinjectors against orientation and temporal changes, mitigating risks associated with handling variability and optimizing supply chain logistics. Future research should extend these findings to longer durations to confirm long-term stability.

Soccer match play induces substantial mechanical, metabolic, inflammatory, and neuromuscular stress, yet post-match monitoring in applied settings often relies on isolated markers, venous sampling, or limited time points. This observational repeated-measures study aimed to describe whether capillary-derived biomarkers, neuromuscular performance, and perceptual measures showed asynchronous recovery during the first 48 h after a standardised soccer match in elite players. Twenty-two elite male outfield soccer players completed a standardised 90 min match. Capillary blood biomarkers, countermovement jump (CMJ), 20 m sprint performance, maximal voluntary contraction (MVC), and delayed onset muscle soreness (DOMS) were assessed before the match, immediately post-match, and at 24 and 48 h post-match. Time effects were analysed using repeated-measures mixed-effects models, and associations between biochemical and functional responses were examined descriptively. Match play induced clear but domain-specific disturbances. IL-6 and cortisol rose rapidly immediately post-match, whereas hsCRP, CK, LDH, myoglobin, and DOMS showed delayed peaks during early recovery. CK, LDH, myoglobin, and soreness remained above baseline at 48 h. CMJ and sprint performance were impaired after the match but largely recovered by 48 h, whereas MVC showed its greatest decrement at 24 h. Exploratory associations indicated that larger muscle damage responses tended to co-occur with greater strength and jump decrements and higher soreness, but these analyses were not causal. Recovery after a standardised elite soccer match was multidimensional and non-synchronous across physiological, neuromuscular, and perceptual domains. A capillary-based, multi-domain assessment strategy may provide a feasible descriptive perspective for field-based observation of post-match fatigue.

Postoperative infectious complications remain a clinically relevant concern after flexible ureteroscopy (FURS), including in patients with negative preoperative bladder urine cultures (UCs). Microbiological discordance between bladder and upper urinary tract samples may contribute to under-recognition of infectious risk. We assessed the clinical value of systematic intraoperative UCs obtained at predefined stages of FURS. We relied on a prospectively collected dataset of 238 consecutive FURS procedures for renal stone disease conducted by a single experienced surgeon performed (2023-2025). UCs were obtained preoperatively and intraoperatively at standardized time points, including bladder urine at the start of the procedure, renal pelvic urine before laser lithotripsy, and renal pelvic urine after lithotripsy. The primary endpoint was postoperative infectious complications, classified as any infection and severe infection (sepsis/septic shock according to international definitions). Associations between culture-positivity patterns and outcomes were evaluated using Fisher's exact test and univariable logistic regression. Among 952 UCs analyzed, 79% were negative overall. A total of 136 patients (57.1%) had negative cultures at all sampling time points. De novo intraoperative culture positivity occurred in 7.1% of patients with negative preoperative UCs. Pre-laser intraoperative UC positivity was significantly associated with postoperative infectious complications, with rates of any infection and severe infection of 27% and 23%, respectively. In univariable analyses, pre-laser intraoperative positivity (either renal pelvic or bladder urine) was associated with an increased risk of any infection (OR 6.14, 95% CI 1.43-24.3; p = 0.01) and severe infection (OR 8.87, 95% CI 1.53-51.8; p = 0.011) in reference to the all negative UC group. In contrast, isolated preoperative or post-laser UC positivity was not significantly associated with infectious outcomes. Intraoperative UCs obtained before laser lithotripsy were significantly associated with postoperative infectious complications following FURS for renal stone disease. Preoperative bladder UCs alone showed limited ability to identify patients at increased infectious risk. As culture results become available 24-48h after surgery, intraoperative pre-laser sampling may represent a valuable microbiological tool to guide timely, culture-informed antibiotic therapy in patients who develop postoperative infectious complications.

Background: Fluoroquinolone prophylaxis during autologous stem cell transplantation (aSCT) reduces the risk of fever but raises the risk of bloodstream infection (BSI) with fluoroquinolone-resistant Enterobacterales (FRE). We performed a prospective cohort study to detect the presence and potential gain or loss of colonic FRE colonization using serial sampling before and after aSCT in a uniform population of patients with a diagnosis of multiple myeloma. Methods: Eligible subjects underwent aSCT after conditioning with dose-intense melphalan, 200 mg/m2. Peri-anal swabs were obtained before aSCT, upon hospital discharge, and 12-16 weeks after transplantation. Samples were cultured in tryptic soy broth supplemented with either ciprofloxacin or ceftriaxone with subsequent plating onto selective chromogenic agar designed to facilitate recovery and differentiation of Enterobacterales. Results: FRE colonization on pre-transplant sampling was detected for 23 of 117 subjects (19.7%) and 29 of 98 (29.6%) subjects at hospital discharge after a course of fluoroquinolone (116/117 subjects) prophylaxis (p < 0.001) and 28 of 92 (30.4%) subjects at 12-16 weeks. Including all three sampling time points, 48 of 117 subjects (41.0%) tested positive for FRE colonization. In total, 58 of the 90 FRE isolates (64.4%) from 48 subjects expressed extended-spectrum beta-lactamase (ESBL). Three FRE-colonized subjects developed FRE BSI. Bloodstream isolates for two subjects were identical to the organisms identified on pre-transplant sampling. Conclusions: We hypothesize that fluoroquinolone prophylaxis of subjects with undetected low levels of FRE colonization allows the expansion of the FRE population, placing subjects at risk of BSI with fluoroquinolone-resistant (and ESBL-expressing) Enterobacterales. Pre-transplant testing for FRE colonization permits patient-specific design of prophylactic and empiric antibiotic regimens.

The intensive use of synthetic fertilizers and pesticides has increased crop productivity but also contributed to soil degradation and biodiversity loss, highlighting the need for more sustainable agricultural strategies. Among emerging solutions, plant growth-promoting rhizobacteria (PGPR), particularly members of the Bacillota phylum, are gaining attention as effective bioinoculants that enhance plant growth and tolerance to biotic and abiotic stresses. However, introduced strains do not function in isolation. They enter complex microbial communities, shaped by plant type and developmental stage, influenced by soil properties and environmental conditions. While the positive effects of PGPR on plant performance are well documented, their impact on indigenous rhizosphere microbiota remains less studied. This review synthesizes current knowledge on how Bacillota-based inoculants influence native microbial communities in cereals, vegetables, orchard crops, and fiber plants. Most studies report shifts toward plant-beneficial taxa and reduced abundance of potential pathogens following Bacillota application. Frequently enriched genera include Bacillus, Pseudomonas, Lysobacter, Sphingomonas, Streptomyces, Azotobacter, Arthrobacter, Pseudarthrobacter, Bradyrhizobium, Devosia, Flavobacterium, Klebsiella, Herbaspirillum, and Rhodanobacter. These changes are often associated with improved plant growth and yield, and stress resilience. However, responses strongly depend on strain, plant and methodological approach. We summarize commonly applied approaches used to assess these interactions. Despite technological advances, limitations remain, such as single time-point sampling, simplified experimental systems, and insufficient integration of inoculant persistence with community analyses. Standardized, multi-site experimental frameworks, with multiple sampling terms are needed to improve predictability and ensure the safe implementation of PGPR-based solutions in sustainable agriculture.

In this study, the results of equine blood cultures sampled during the period from 2022 to 2024 were evaluated. Samples were submitted as routine samples by veterinary practices to a veterinary diagnostic laboratory in Germany. The study focused mainly on the differentiation of the bacterial isolates. In order to place the results in a clinical context, the study was complemented by a questionnaire sent to the practices enquiring for sepsis-associated clinical signs. The study was carried out retrospectively. A total of 199 blood culture samples submitted by practices located in Germany and other European countries were included into the evaluation. A questionnaire asking for the reason of the individual blood collection and possible presence of sepsis-associated clinical signs was sent to the relevant practices and the returning responses were analyzed. 50.3% (n=100) of the blood culture samples showed a positive result, divided into 71% monomicrobial and 29% polymicrobial samples. Staphylococci were most frequently detected (30.2%), followed by bacteria of the genus Enterobacterales (18.9%). Nearly half of the horses (41.3%) was pretreated with antibiotics at the time of sample collection. Respiratory infection was the most frequently listed reason (17.4%) for initiation of a blood culture analysis. The results of the present study are comparable to previous studies. The isolates were not classified into clinically relevant pathogens and contaminants, as for the majority of patients only one sampling timepoint was available, making any isolate a potential causative agent for bacteremia. Sepsis is not a specific condition, but is based on the clinical findings and the result of the performed blood culture analysis. Considering the duration of a bacteriological culture procedure as well as the commonly critical condition of the patient, availability of data concerning the most frequently occurring pathogens is essential in rapidly initiating adequate treatment.

MRD-2 in the GHSG HD21 trial assessed by a validated circulating tumor DNA sequencing assay.

Recent studies have applied machine learning (ML)-based limited sampling strategies (LSS) to predict drug exposure (AUC), achieving low prediction error and performance comparable to or better than multiple linear regression and population pharmacokinetics LSS. This study aimed to develop and validate a machine learning-based limited sampling strategy capable of predicting raltegravir (RAL) exposure. Four machine learning algorithms (XGBoost, Random Forest, GLMNet, and SVM) were trained using pharmacokinetic profiles generated via Monte Carlo simulation from a population pharmacokinetic (POPPK) model. Data were divided into training (75%) and test (25%) sets. All possible combinations of sampling times, pairs and triplets, in steady-state, up to 12 h post-dose were evaluated. Model performance was assessed by the lowest root mean square error (RMSE) in the cross-validation, and the best performing model was evaluated in the test set and externally validated using simulated PK profiles from an independent POPPK model and patient data from a clinical study. XGBoost trained with concentrations at 0.5, 2, and 4 h showed the best predictive performance. The model achieved excellent accuracy in the test set (bias/RMSE: 0.8%/8.7%) and in the independent simulation (1.9%/14.3%). Performance decreased in real patient data (5.0%/24.1%), highlighting the need for caution when extrapolating predictions to populations whose characteristics differ from those represented in the training datasets. A machine learning model using only three sampling timepoints has been developed and validated in different datasets, enabling accurate estimation of RAL AUC₀-₁₂. This approach provides a tool for pharmacokinetic and PK/PD studies and reduces intensive sampling need in clinical settings.

Transcriptomic analyses reveal the status of cells, tissues, or organisms, across states of health and disease. RNA velocity adds a temporal dimension to single cell analyses, predicting future transcriptomic and phenotypic states, based on the current spliced and unspliced mRNA of each cell. We hypothesized that RNA velocity could be adapted to predict future clinical state of individuals with acute and chronic illnesses, using their whole-blood transcriptomes. We developed VeloCD, a method for quantitative prediction of transitions in clinical state from a single time-point RNA sample. This predicts transcriptomic trajectories and future infection status in influenza A and SARS-CoV-2 controlled human infection studies, which are consistent with trajectories in naturally acquired infections. In HIV-TB coinfected individuals, VeloCD predicts the onset of immune reconstitution inflammatory syndrome. In individuals receiving biological therapy for inflammatory bowel disease, whole blood RNA velocity after the first dose of treatment indicates whether remission will be achieved by the end of the treatment course. In a multinational observational study of acutely unwell febrile children, VeloCD predicts those with greatest medical care requirements. Our results demonstrate proof-of-concept for the use of RNA velocity to predict trajectories of human diseases.

Abstract Wastewater represents a complex, unstructured mixture of biological material contributed by humans, animals, and other organisms, making it a valuable resource for pathogen surveillance. This study presents a comprehensive capture-probe-based virome analysis conducted over 1 year across four metropolitan cities, with samples collected fortnightly from 24 different sites. We identified over 170 DNA and 135 RNA viral species, including viral strains associated with multiple hosts. Notably, we detected 107 DNA viruses and 495 RNA viral genomes/segments with over 90% genome coverage. Beyond detection, we explored temporal patterns, revealing that viral families such as Astroviridae, Orthomyxoviridae, and Picornaviridae exhibited seasonal trends, while Adenoviridae and Sedoreoviridae were consistently detected throughout the year. Shannon diversity was higher and more variable at several sites in Ahmedabad, likely due to its larger population size, landscape, and heavy footfall. Bray-Curtis dissimilarity analysis showed significant variation in viral communities across cities, sites, and sampling time points. The prevalence of SARS-CoV-2 and Rotavirus A from NGS showed a significant positive correlation with digital PCR (dPCR) quantification ( p &lt; 0.05). Additionally, trends in Hepatitis A virus abundance aligned with monthly reported cases, further validating the reliability of sequencing-based surveillance.