Search Results (649)

This study aimed to evaluate daily patterns of hourly milking frequency (MF) in dairy cows milked with an automatic milking system (AMSs), considering the effects of season, parity order (PO), days in milk (DIM), and milk yield (MY). A retrospective longitudinal study was conducted on a commercial dairy farm in southern Brazil over one year using data from 130 Holstein cows and 94,611 milking events. MF data were analyzed using general linear models. Overall, hourly MF followed a consistent daily pattern, with peaks between 4:00 and 11:00 a.m. and between 2:00 and 6:00 p.m., regardless of season, PO, DIM, or MY category. MF was higher in primiparous (2.84/day, p = 0.0013), early-lactation (<106 DIM; 3.00/day, p < 0.0001), and high-yielding cows (≥45 L/day; 3.09/day, p < 0.0001). High-yielding cows also showed sustained milking activity into the late nighttime. Although seasonal and individual factors significantly affected MF, they had limited influence on the overall daily distribution of milkings. These results suggest stable behavioral patterns within the specific AMS management conditions observed in this study and suggest that adjusting milking permissions and feeding strategies based on cow characteristics may improve system efficiency. Full article

Due to the feeding system (high-concentrate diet) during the early lactation stage, ruminal pH in dairy cows follows a diurnal pattern and can remain below the critical level of 5.5 for extended periods of the day. This study aimed to evaluate the effect of low ruminal pH on blood concentrations of certain macrominerals, trace minerals, and fat-soluble vitamins and on the oxidative status of dairy cows during the first half of lactation. Fifty-three randomly selected lactating Holstein cows were used; blood and ruminal fluid samples were collected from all cows on days 30, 90 and 150 of lactation. Blood samples were obtained via coccygeal venipuncture, while the ruminal fluid was obtained by rumenocentesis and the pH was measured immediately after collection. Using a threshold pH of 5.5, samples were classified as normal (pH > 5.5) or low pH (pH ≤ 5.5). Serum concentrations of Ca, Mg, K, Cr, Mn, Zn, Se, and vitamins A, D3, E, and K were not significantly affected by ruminal pH, either by days in milk or by their interaction (p > 0.05). Plasma malondialdehyde and reduced glutathione followed the same trend (p > 0.05). Copper concentration was significantly higher (p < 0.05), and Fe concentration tended to be higher in cows with low pH compared to those with normal pH (p = 0.052). On day 150 of lactation, Cu, Fe, and Co concentrations were significantly higher in low-pH cows compared to normal-pH cows (p < 0.05). Low ruminal pH is associated with significant changes in serum concentrations of copper, iron, and cobalt but has no significant effect on the oxidative status of the animals or on the serum concentrations of the macro elements and fat-soluble vitamins studied. Full article

Oxidative and glycolytic metabolism produce energy in the form of ATP and produce intermediates for biomass production. Oxidative metabolism predominates under normoxic conditions and in quiescent or slowly proliferating cells. On the other hand, under hypoxic or pseudohypoxic conditions and in rapidly proliferating cells, glycolysis becomes the predominant pathway. The balance between oxidative and glycolytic metabolism is finely tuned in physiological conditions and becomes dysregulated in many pathological conditions, most notably cancer. In this article we summarize the evidence that has been gathered over the last few years on the mechanisms underlying this balance and the consequences of their dysregulation. We discuss first the non-metabolic factors (mitochondria, cell cycle, cell type, tissue type), then molecules that are at the intersection between glycolytic and oxidative metabolism and those molecules that are inherent to oxidative or glycolytic metabolism that affect the equilibrium between the two energy-producing pathways. Eventually, we discuss pharmacologic or genetic means that allow manipulating this equilibrium. As will be seen, lactic acidosis has taken center stage in this field and lactate has been shown to fuel oxidative metabolism. This suggests that if glycolytic metabolism predominates, as has often been shown in cancer, mechanisms come into work that reestablish a metabolic heterogeneity. Thus, while one pathway may be predominant over the other, it seems as if fail-safe mechanisms are at work that avoid the possibility that it becomes the only energy-producing pathway. Eventually, we discuss possible therapeutic consequences that may derive from this expanding knowledge, in particular, as regards tumor therapy. Full article

This review aimed to be comprehensive and to critically analyze the factors that may affect the macronutrient and energy content of breast milk. Systematic reviews were prioritized, even though other types of literature reviews on the subject, as well as studies not included in these reviews, were included. Reported factors that potentially affect the macronutrient and energy content of breast milk comprise: maternal factors, such as age, nutritional status, dietary intake, smoking habits, lactation stage, circadian rhythmicity, and the use of galactagogues; obstetrical factors, such as parity, preterm delivery, multiple pregnancies, labor and delivery, and pregnancy morbidities including intrauterine growth restriction, hypertensive disorders, and gestational diabetes mellitus; and newborn factors, including sexual dimorphism, and anthropometry at birth. Some factors underwent a less robust assessment, while others underwent a more in-depth analysis. For example, the milk from overweight and obese mothers has been reported to be richer in energy and fat. A progressive decrease in protein content and an increase in fat content was described over time during lactation. The milk from mothers with hypertensive disorders may have a higher protein content. Higher protein and energy content has been found in early milk from mothers who delivered prematurely. Full article

Background/Objective: The hepatitis B virus (HBV) can cause chronic hepatitis B (CHB), which can rapidly progress into fatal liver cirrhosis (CHB-LC) and hepatocellular carcinoma (CHB-HCC). Methods: In this study, we investigated metabolites associated with distinct clinical stages of HBV infection for the identification of stage-specific serum metabolite biomarkers using ¹H-NMR-based metabolomics. Results: A total of 64 serum metabolites were identified, among which six core discriminatory metabolites, namely isoleucine, tryptophan, histamine (for CHB), and pyruvate, TMAO, lactate (for CHB-HCC), were consistently significant across univariate and multivariate statistical analyses, including ANOVA with FDR, OPLS-DA, and VIP scoring. These metabolites were closely linked to key metabolic pathways, such as propanoate metabolism, pyruvate metabolism, and the Warburg effect. Conclusions: The findings suggest that these six core metabolites serve as potential stage-specific biomarkers for CHB, CHB-LC, and CHB-HCC, respectively, and offer a foundation for the future development of metabolomics-based diagnostic and therapeutic strategies. Full article

Measuring methane (CH₄) emissions from dairy systems is crucial for advancing sustainable agricultural practices aimed at mitigating climate change. However, current CH₄ measurement techniques are primarily designed for controlled research settings and are not readily scalable to diverse production environments. Thus, there is a need to develop accessible, production-level methods for estimating CH₄ emissions. This review examines the relationship between enteric CH₄ emissions and milk fatty acid (FA) composition, highlights key FA groups with potential as biomarkers for indirect CH₄ estimation, and outlines critical factors of predictive model development. Several milk FAs exhibit strong and consistent correlations to CH₄ emissions, supporting their utility as predictive biomarkers. Saturated and branched-chain FAs are generally positively associated with CH₄ emissions, while unsaturated FAs, including linolenic acid, conjugated linoleic acids, and odd-chain FAs, are typically negatively associated. Variability in the strength and direction of correlations across studies is often attributable to differences in diet or lactation stage. Similarly, differences in experimental design, data processing, and model development contribute to much of the variation observed in predictive equations across studies. Future research should aim to (1) identify milk FAs that consistently correlate with CH₄ emissions regardless of diet, (2) develop robust and standardized prediction models, and (3) prioritize the external validation of prediction models across herds and production systems. Full article

Although age-related changes in the gut microbiome of pigs have been extensively studied, the dynamic patterns of fecal microbiota and SCFAs during the gestation-to-weaning period in sows remain poorly characterized. We aim to characterize the changes in fecal microbiota and SCFAs from pregnancy to weaning, and to investigate their associations with maternal weight gain during gestation. We systematically collected 100 fecal samples at four time points (day 30 of pregnancy (T1), 1–2 days before delivery (T2), day 10 after delivery (T3), and day 21 of weaning stage (T3)), and measured the body weight of sows at T1 (132 kg ± 10.8) and T2 (205 kg ± 12.1). The primary nutrient components of the diets during the gestation and lactation periods are summarized. All fecal samples were subjected to 16S rRNA gene sequencing. We found that a high proportion of crude fiber (bran) is a key feature of the gestation diet, which may affect enterotype shifts and gut microbial composition. Sows fed a high-fiber diet showed significant enrichment of gut microbiota, including genera such as Prevotellaceae_UCG-003, Prevotellaceae_NK3B31_group, and Prevotella_9 during the gestational period (LDA score > 2). Moreover, Eubacterium_coprostanoligenes_group (average relative abundance: 5.5%) and Lachnospiraceae_NK4A136_group (average relative abundance: 2.5%) were the dominant bacteria during the lactation stage. Fecal propionate and butyrate levels were lowest in late gestation, and propionate negatively and acetate positively correlated with body weight change (p < 0.05). Additionally, certain Prevotella taxa were associated with arachidonic acid metabolism and acetate production (p < 0.05). Our study identified key microbial communities across four stages from gestation to weaning and revealed that dietary patterns can shape the sow gut microbiota. Furthermore, we observed significant correlations between SCFAs and body weight change during pregnancy. These findings provide a scientific basis and theoretical support for future strategies aimed at modulating gut microbiota and targeting SCFAs to improve maternal health and productivity throughout the gestation-to-weaning period. Full article

Introduction: Two-stage revision with an antibiotic-loaded, temporary static cement spacer is a common treatment for periprosthetic joint infection (PJI) of the knee. However, limited data exists on in vivo antibiotic elution kinetics after spacer implantation. This pilot study uses the technique of microdialysis (MD) to collect intra-articular knee samples. The aim was to evaluate MD as an intra-articular sampling method to detect spacer-eluted antibiotics within 72 h after surgery and to determine whether they show specific elution kinetics. Methods: Ten patients (six male, four female; age median 71.5 years) undergoing two-stage revision for knee PJI were included. A MD catheter was inserted into the joint during explantation of the infected inlying implant and implantation of a custom-made static spacer coated with COPAL cement (0.5 g gentamicin (G) and 2 g vancomycin (V)). Over 72 h postoperatively, samples were collected and analyzed for spacer-eluted antibiotics, intravenously administered antibiotics (e.g., cefazolin and cefuroxime), metabolic markers (glucose and lactate), and Interleukin-6 (IL-6). Local and systemic levels were compared. Results: All catheters were positioned successfully and well tolerated for 72 h. Antibiotic concentrations in MD samples peaked within the first 24 h (G: median 9.55 µg/mL [95% CI: 0.4–17.36]; V: 37.57 µg/mL [95% CI: 3.26–81.6]) and decreased significantly over 72 h (for both p < 0.05, G: 4.27 µg/mL [95% CI: 2.26–7.2]; V: 9.69 µg/mL [95% CI: 3.86–24]). MD concentrations consistently exceeded blood levels (p < 0.05), while intravenously administered antibiotics showed higher blood concentrations. Glucose in MD samples decreased from 17.71 mg/dL to 0.89 mg/dL (p < 0.05). IL-6 and lactate concentrations showed no difference between MD and blood samples. Conclusions: Monitoring antibiotics eluted by a static spacer with intra-articular MD for 72 h is feasible. Gentamicin and vancomycin levels remained above the minimal inhibitory concentration. Differentiating infection from surgical response using metabolic and immunological markers remains challenging. Prolonged in vivo studies with MD are required to evaluate extended antibiotic release in two-stage exchanges. Full article

This study aimed to investigate the tyrosine kinase inhibitor Nintedanib and its potential role in reversing epithelial–mesenchymal transition (EMT) induced by transforming growth factor beta 2 (TGF-β2) in retinal pigment epithelial (RPE) cells, along with its therapeutic potential using a mouse model of subretinal fibrosis. We hypothesized that the blockade of angiogenesis promoting and fibrosis inducing signaling using the receptor tyrosine kinase inhibitor Nintedanib (OfevTM) can prevent or reverse EMT both in vitro and in our in vivo model of subretinal fibrosis. Primary human retinal pigment epithelial cells (phRPE) and adult retinal pigment epithelial cell line (ARPE-19) cells were treated with TGF-β210 ng/mL for two days followed by four days of Nintedanib (1 µM) incubation. Epithelial and mesenchymal phenotypes were assessed by morphological examination, quantitative real-time polymerase chain reaction(qPCR) (ZO-1, Acta2, FN, and Vim), and immunocytochemistry (ZO-1, vimentin, fibronectin, and αSMA). Metabolites were measured using luciferase-based assays. Extracellular acidification and oxygen consumption rates were measured using the Seahorse XF system. Metabolic-related genes (GLUT1, HK2, PFKFB3, CS, LDHA, LDHB) were evaluated by qPCR. A model of subretinal fibrosis using the two-stage laser-induced method in C57BL/6J mice assessed Nintedanib’s therapeutic potential. Fibro-vascular lesions were examined 10 days later via fluorescence angiography and immunohistochemistry. Both primary and ARPE-19 RPE stimulated with TGF-β2 upregulated expression of fibronectin, αSMA, and vimentin, and downregulation of ZO-1, consistent with morphological changes (i.e., elongation). Glucose consumption, lactate production, and glycolytic reserve were significantly increased in TGF-β2-treated cells, with upregulation of glycolysis-related genes (GLUT1, HK2, PFKFB3, CS). Nintedanib treatment reversed TGF-β2-induced EMT signatures, down-regulated glycolytic-related genes, and normalized glycolysis. Nintedanib intravitreal injection significantly reduced collagen-1+ fibrotic lesion size and Isolectin B4+ neovascularization and reduced vascular leakage in the two-stage laser-induced model of subretinal fibrosis. Nintedanib can induce Mesenchymal-to-Epithelial Transition (MET) in RPE cells and reduce subretinal fibrosis through metabolic reprogramming. Nintedanib can therefore potentially be repurposed to treat retinal fibrosis. Full article

Esophageal adenocarcinoma (EAC) is a highly aggressive malignancy with rising incidence and poor prognosis. TP53, previously identified as the most frequently mutated gene in EAC in our studies, plays a central role in tumor suppression and regulation. However, the metabolic consequences of TP53 mutations in EAC remain largely uncharacterized. We metabolically profiled three TP53-mutant EAC cell models (OE33, OE19, and FLO1) representing progressive stages of tumor differentiation and harboring distinct TP53 alterations. Our analyses revealed different metabolic phenotypes associated with TP53 status. OE33 cells predominantly use glycolytic metabolism but display limited adaptability to environmental changes, possibly due to a higher differentiation state. FLO1 cells exhibit a strong glycolytic dependence, elevated lactate production, and robust proliferation under acidic conditions, consistent with an aggressive and metastatic phenotype. OE19 cells preferentially utilize oxidative phosphorylation, demonstrated by resilience to glucose and glutamine deprivation, and ROS accumulation. These findings highlight the metabolic plasticity of EAC and suggest that TP53 mutation type might influence bioenergetic dependencies. Targeting these metabolic vulnerabilities may offer novel therapeutic avenues for personalized treatment in EAC. Full article

This study investigated the effect of different farming systems and lactation stages on the physicochemical characteristics, somatic cell count (SCC), and total bacterial count (TBC) of milk from Skopelos goats. This study was conducted over two consecutive lactation periods on two commercial farms in Greece, an extensive system on Skopelos Island and an intensive system in the Attica region, involving 237 goats of shared genetic background, thereby minimizing genetic variability and strengthening the validity of the comparisons between the production systems. Higher milk yields were observed in the extensive system (0.98 vs. 0.85 kg/day), while milk from this system also had a higher protein (3.57% vs. 3.47%; p < 0.001) and casein content (2.72% vs. 2.57%; p < 0.001), which are traits favorable for cheese production. Fat content peaked during mid-lactation (4.83%; p < 0.05) and remained unaffected by the farming system. Lactose declined from early (4.74%) to late lactation (4.42%; p < 0.001). Both SCC and TBC were significantly elevated in the extensive system (p < 0.001), possibly due to hand milking, environmental exposure, and less-controlled hygiene conditions. These findings highlight a trade-off between the nutritional advantages of extensive systems and challenges related to milk hygiene. A balanced approach, optimizing both quality and sustainability, is recommended. Full article

Background/Objectives: Pancreatic cancer (PC) is an aggressive malignancy with a poor prognosis, frequently diagnosed at a metastatic stage. The identification of accessible, cost-effective prognostic biomarkers is critical for optimizing treatment strategies. The Endothelial Activation and Stress Index (EASIX), calculated using lactate dehydrogenase (LDH), creatinine, and platelet count, reflects endothelial dysfunction and has shown prognostic value in hematological cancers. However, its utility in metastatic PC remains unexplored. This study is the first to evaluate the prognostic significance of the EASIX in patients with metastatic PC receiving first-line FOLFIRINOX chemotherapy. Methods: This retrospective cohort study analyzed 204 patients diagnosed with metastatic pancreatic adenocarcinoma at Istanbul Training and Research Hospital between 2020 and 2025. All patients received FOLFIRINOX as first-line therapy. EASIX was calculated as LDH (U/L) × creatinine (mg/dL)/platelet count (10⁹/L). A cut-off value of 1.33 was used to stratify patients into low and high EASIX groups. Overall survival (OS) was assessed using Kaplan–Meier analysis and compared with the log-rank test. Results: The mean patient age was 63.0 ± 9.4 years; 61.8% were male. There were no significant differences in baseline characteristics between groups. Patients with EASIX ≥ 1.33 had significantly lower platelet counts and higher LDH and creatinine levels. Median OS was 14 months for EASIX < 1.33 and 8 months for EASIX ≥ 1.33 (p < 0.001). Conclusions: EASIX is a simple, inexpensive prognostic marker associated with overall survival in metastatic PC. Its integration into clinical practice may facilitate early risk stratification. Further prospective studies are needed to confirm its prognostic utility. Full article

Cryptosporidium spp. are protozoan parasites that cause cryptosporidiosis, an enteric disease that can affect a wide range of vertebrate hosts. Pigs play a potential role in the transmission of Cryptosporidium spp. to humans, although infections are most often subclinical. This study aimed to assess the occurrence and molecular characterization of Cryptosporidium spp. in swine farms located in Aragón, northeastern Spain. Fecal samples (n = 72) were collected from 10 breeding farms, encompassing various production stages (lactation, nursery, fattening, and/or wean-to-finish). Data regarding the type of production system (two- or three-stage), production stages, and farming facilities (the type of flooring and water source) associated with the parasite presence were also analyzed using a questionnaire. The results showed that Cryptosporidium spp. were more frequently detected in fecal samples originating from three-stage production systems (21.9%) compared to two-stage systems (12.5%). Samples from the fattening stage exhibited the highest positivity rate and estimated oocyst count (3.0 oocyst/microscopic field). Furthermore, the molecular characterization of Cryptosporidium spp. revealed the circulation of multiple species both among farms and within the same pig production flow, with Cryptosporidium scrofarum being the most prevalent species (7/72; 9.7%), followed by Cryptosporidium suis (1/72; 1.4%). These findings underscore the importance of the surveillance and molecular characterization of Cryptosporidium spp. for controlling infections in pigs, considering the potential for the zoonotic transmission of this parasite to humans. Full article

Objectives: The present study aimed to determine the reference intervals for complete blood count and total protein parameters in Greek indigenous Capra prisca goats and to evaluate their associations with parasitic burden, age and reproductive stage. Methods: Two-hundred clinically health goats were grouped by parasite status (gastrointestinal nematodes, Eimeria spp., and lungworm infection), age (3–6-month-old growing kids; lactating non-pregnant goats ≤ 3 or >3 years old) and reproductive stage (non-lactating pregnant goats; lactating non-pregnant goats). Blood samples were analyzed for erythrogram, leukogram and megakaryocytic parameters using an automated analyzer and manual blood smears. Total plasma proteins were measured using refractometry. Results: Gastrointestinal nematode-infected animals (>300 eggs per gram of feces) were associated with a significant reduction in red blood cell counts and hematocrit estimation, and an increase in mean corpuscular hemoglobin and mean corpuscular hemoglobin concentrations, while lungworm-infected animals were associated with decreased red blood cells, red cell distribution width and neutrophils, and increased lymphocytes compared to non-infected animals. Eimeria spp. affected only basophils in growing kids. Age influenced all erythrocytic and leukocytic parameters (apart from neutrophils and monocytes), as well as all megakaryocytic parameters and total proteins, with younger animals showing higher red and white blood cell counts and platelets compared to adults. Pregnant does had elevated hemoglobin, hematocrit, neutrophils and monocytes compared with lactating non-pregnant does. Conclusions: The calculated 95% reference intervals for our demographic groups of animals provide a useful diagnostic framework for assessing Capra prisca health in Greek goat farming. Full article

Background: Septic shock is a heterogeneous syndrome with diverse clinical presentations and pathophysiology, yet current management guidelines largely treat it as a homogenous entity. Early risk stratification relies on lactate and different predictive scoring systems, which may not capture the underlying heterogeneity in host responses. Aim: To identify discrete subphenotypes of septic shock using admission demographics and laboratory parameters, and to evaluate their relationship with in-hospital outcomes. Methods: We conducted a retrospective multicenter cohort study of 10,462 adult patients with ICD-10-defined septic shock admitted to intensive care units between 2014 and 2015. We used Two-Step Cluster Analysis using log-likelihood distance and the Bayesian Information Criterion to identify two distinct phenotypes. We compared clusters on baseline characteristics, in-hospital outcomes including mortality, days on mechanical ventilation, vasopressor use, acute kidney injury (AKI), AKI requiring renal replacement therapy (RRT), and ICU and hospital lengths of stay. Results: We identified two clusters (Cluster 1, n = 5355 and Cluster 2, n = 5107) in our study. Cluster 1 showed greater biochemical severity at presentation, including higher median lactate (2.40 vs. 2.20 mmol L⁻¹; p < 0.001), serum creatinine (1.39 vs. 1.20 mg dL⁻¹; p < 0.001), blood urea nitrogen (28 vs. 25 mg dL⁻¹; p < 0.001), and neutrophil-to-lymphocyte ratio (11.12 vs. 10.38; p < 0.001), and a higher mean SOFA score (7.05 ± 3.85 vs. 6.76 ± 3.87; p < 0.001). Despite this, Cluster 1 required mechanical ventilation more frequently (46.1% vs. 42.2%; p < 0.001) and had a higher incidence of AKI (58.1% vs. 55.6%; p = 0.009), including more stage 3 AKI (17.2% vs. 15.2%; p < 0.001) and dialysis (6.6% vs. 5.2%; p = 0.005), yet experienced similar in-hospital mortality (15.4% vs. 15.8%; p = 0.615) and comparable ICU (2.18 vs. 2.26 days; p = 0.254) and hospital lengths of stay (6.63 vs. 6.80 days; p = 0.251). Conclusions: Two septic shock phenotypes were identified, one with marked early organ dysfunction (Cluster 1) and another with milder initial derangements (Cluster 2), yet both showed convergent short-term mortality and lengths of stay despite divergent support needs. These results challenge reliance on single-parameter severity markers and underscore the need for phenotype-guided risk stratification and personalized management strategies in septic shock. Full article