Search Results (159)

Introduction: Intravenous push (IVP) beta-lactam antibiotics have been adopted during parenteral solution shortages to conserve resources. Data evaluating the safety and efficacy of cefepime administered IVP versus intravenous piggyback (IVPB) infusion in Gram-negative bacteremia remain limited. We compared clinical outcomes of cefepime administered IVP versus IVPB in hospitalized patients with Gram-negative bacteremia. Methods: This was a retrospective cohort study across a five-hospital health system from 1 January 2014 through 31 December 2021. Adults receiving cefepime for Gram-negative bacteremia were included. The primary outcome was a tailored desirability of outcome ranking (DOOR) composite assessed through 30 days or hospital discharge, integrating clinical cure and cefepime-associated neurologic adverse effects. Clinical cure was defined as absence of recurrent bacteremia with the index pathogen after 48 h, no antibiotic escalation, and no in-hospital mortality. Results: A total of 254 met the inclusion criteria (127 IVPB; 127 IVP). Baseline severity was similar between groups. The primary outcome assessed by DOOR revealed no difference between IVPB and IVP groups (p = 0.656). Vasopressor support during therapy was more frequent in the IVP group (22.0% vs. 10.2%, p = 0.011), and median hospital length of stay was longer (10 vs. 7 days, p = 0.020). No differences were noted in other endpoints. General ward admission (OIR [aOR] 2.563, 95% CI 1.271–5.168; p = 0.009) and genitourinary source of bacteremia (aOR 3.398, 95% CI 1.509–7.652; p = 0.003) independently predicted clinical cure. Conclusions: In patients with Gram-negative bacteremia, cefepime administered IVP demonstrated similar safety and efficacy to IVPB infusion. Full article

Feline calicivirus (FCV) is a major pathogen that threatens feline health worldwide. Its global prevalence, extensive genetic variability, and limited cross-protection among strains present significant challenges for vaccine development. In this study, an infectious clone of the FCV-GDJM202201 strain was constructed using the eukaryotic expression plasmid pcDNA3.1 under the control of the cytomegalovirus (CMV) promoter. The rescued virus, rGDJM-A4822T, exhibited growth kinetics comparable to those of the parental strain in vitro. Subsequently, two recombinant viruses, rGDJM-VP1_JL and rGDJM-VP1_SH, were generated by replacing the VP1 gene in the GDJM202201 backbone with those from heterologous FCV strains. Notably, these recombinant viruses exhibited reduced viral titers compared to rGDJM-A4822T. Finally, neutralization assays revealed differential neutralizing antibody titers among the recombinant FCVs, with rGDJM-A4822T inducing higher neutralizing antibody titers and cross-neutralizing activity. Collectively, this study establishes an FCV infectious clone that can be used to rescue recombinant viruses carrying heterologous VP1 proteins and to evaluate neutralizing antibody responses. Full article

In this study, we investigated the long-term evolutionary dynamics of human norovirus GII.17[P17] using the RNA-dependent RNA polymerase (RdRp) region and the VP1 capsid gene, integrating phylogenetics, time-scaled inference, phylodynamics, and structure-based analyses. Maximum-likelihood phylogenies of both genomic regions consistently resolved four major clades (Clades 1–4). VP1 patristic-distance distributions indicated higher within-clade diversity in the phylogenetically basal Clades 1 and 3, whereas Clades 2 and 4 showed lower diversity, consistent with recent demographic expansion. Similarity-plot analysis identified pronounced variability in the VP1 P2 domain, while the S and P1 domains remained comparatively conserved, supporting P2 as the primary hotspot of diversification. Bayesian time-scaled analyses estimated the most recent common ancestor around 1993 (VP1) and 2000 (RdRp) and revealed two major lineages (Clade 1/2 and Clade 3/4), with the split between Clades 3 and 4 occurring around 2016–2017. Bayesian skyline plots showed a marked increase in effective population size after 2013, and substitution-rate estimates indicated faster evolution in VP1 than in RdRp, with higher VP1 rates in the Clade 3/4 lineage than in Clade 1/2. Capsid dimer modeling further mapped high-confidence conformational B-cell epitopes and positively selected residues predominantly to the distal surface of P2, with broadly conserved spatial patterns across clades. Compared with the Clade 1 reference (Kawasaki323), Clade 2 accumulated numerous P2 substitutions, whereas Clades 3 and 4 retained fewer changes and remained closer to Clade 1 at the amino-acid level. Together, these results suggest lineage turnover within GII.17[P17] driven by constrained diversification at the P2 surface, potentially contributing to the recent predominance of the Clade 3/4 lineage. Full article

High-efficiency Ovum Pick-Up (OPU) and in vitro embryo production (IVP) are critical for the genetic improvement of high-value Wagyu cattle. However, oxidative stress and mitochondrial dysfunction during oocyte maturation remain major bottlenecks limiting blastocyst yield. This study investigated the role of inhibin in Wagyu oocyte competence through two independent proof-of-concept approaches. In the in vivo active immunization model, thirty Wagyu donors were immunized with a recombinant inhibin protein (INHA group), resulting in a significant increase in the number of recovered cumulus–oocyte complexes (COCs) (461 vs. 279, p < 0.05) and the proportion of high-quality oocytes compared to controls. Oocytes from the INHA group exhibited improved cytoplasmic maturation and mitochondrial function, characterized by higher membrane potential (ΔΨm, JC-1 ratio: 1.55 ± 0.06 vs. 0.83 ± 0.08, p < 0.05), elevated ATP content (2.35 ± 0.07 vs. 1.63 ± 0.03 pmol/oocyte, p < 0.05), and increased NADPH levels. Furthermore, the INHA group showed significantly reduced reactive oxygen species (ROS) accumulation and an increased GSH/GSSG ratio (8.48 ± 0.18 vs. 6.25 ± 0.09, p < 0.05), indicating restored redox homeostasis. Independently, in the in vitro anti-inhibin antibody (AIA) supplementation model, AIA supplementation during oocyte maturation significantly improved the nuclear maturation rate (92.96% ± 1.04%), blastocyst formation rate (56.63% ± 2.36%), and total cell number compared to controls (p < 0.05). Notably, AIA-derived blastocysts achieved a significantly higher pregnancy rate (78.65% ± 1.57%) following transfer. Collectively, these findings demonstrate that targeting inhibin mitigates oxidative injury and stabilizes mitochondrial bioenergetics, providing two distinct, physiology-based strategies for optimizing Wagyu oocyte yield and embryo production. Full article

In vitro embryo production (IVP) has emerged as a crucial tool in assisted reproduction and animal biotechnology. A key factor in this process is in vitro oocyte maturation (IVM), a critical process preceding fertilization that directly influences embryo quality. FLI supplementation, composed of fibroblast growth factor (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor 1 (IGF1), has been shown to facilitate the IVM process to mimic essential aspects of in vivo oocyte development, and therefore, promote higher rates of oocyte maturation, embryonic viability, blastocyst formation, and improve the number of live animals born after embryo transfer. Individually or together, these components participate in signaling pathways that are crucial for improving oocyte competence and early embryo development. This review highlights the individual and combined roles of FGF2, LIF, and IGF1 in maturation and embryo culture medium, their influence on subsequent embryonic development, and their signaling pathways. Additionally, the incorporation of antioxidants and amino acids as supplementary components in combination with FLI is explored as a strategy to mitigate oxidative stress and enhance metabolic support during IVM and embryo culture. Together, these elements can significantly improve IVP outcomes, providing a potential pathway for optimizing the efficiency of embryo production in various species. Full article

In classical explicit Runge–Kutta methods for solving initial value problems (IVPs) of the form $y^{\prime }\left(x\right)=f(x,y),y\left(x_0\right)=y_0$, the first stage is typically given by evaluating the right-hand side at the initial point, i.e., $f_1=f(x_0,y_0)$. However, this approach becomes inefficient or even ill-posed when the $f(x_0,y_0)$ exhibits a singularity at $x_0$, as is common in many physically motivated problems such as the Lane–Emden equation or Thomas–Fermi model. To address this issue, we propose an alternative approach that was originally introduced by Oliver for low-order methods. In this formulation, the first stage is shifted away from the singular point and is instead evaluated at a shifted location: $f_1=f(x_0+c_1\tau ,y_0)$, where $\tau$ is the step size and $c_1\ne 0$ is a nonzero coefficient. This allows the method to bypass the singularity while preserving consistency with the IVP. We derive the corresponding order conditions for algebraic order six and construct an eight-stage scheme satisfying these constraints. The resulting method demonstrates significantly improved efficiency when applied to problems with initial-point singularities, outperforming classical Runge–Kutta pairs of orders $6\left(5\right)$ and even $8\left(7\right)$. Full article

The formation and expansion capacity of blastocysts plays a very important role in successful implantation. During mammalian embryo development derived from in vitro production (IVP), early embryos are highly susceptible to various cellular stresses, including endoplasmic reticulum (ER) stress, which has been identified in IVP embryos, suggesting that ER stress modulation is crucial for IVP embryo development. In this study, we aimed to evaluate the influences of ER stress on preimplantation embryos during blastocyst formation and expansion potential in pigs. Tunicamycin (TM), an ER stress inducer, was employed in porcine embryos, significantly increasing the mRNA levels of ER stress-related markers ATF6, CHOP, and GRP78. When one-cell embryos were cultured in the presence of TM, the blastocyst formation and diameter (reflecting the blastocyst expansion capacity) were significantly inhibited in a dose-dependent manner. When morula-stage porcine embryos were cultured in TM, the blastocyst formation rate, blastocyst diameter, total cells and EdU-positive cell numbers were significantly lower than the TM-free control group. TM reduced the potential of blastocoel recovery (ex-expansion) in blastocysts collapsed by cytochalasin D and impeded blastocyst expansion. In addition, TM reduced the mRNA levels of CDH1 and TJP1 and affected the normal expression pattern of E-cadherin, Oct4, Sox2 and Cdx2 in porcine blastocysts. Taken together, these findings suggest that TM treatment during embryo development in vitro interferes with the formation and expansion capacity of the blastocoel in pigs. Full article

In bovine embryo transfer (ET) using in vitro-produced (IVP) embryos, recipient factors and embryo grade are well-established predictors of pregnancy success, but the impact of the laboratory-level developmental competence of IVP embryos remains insufficiently characterized. This retrospective study evaluated factors affecting pregnancy rates following the transfer of IVP beef embryos to dairy recipients. Medical records from 462 ETs were analyzed across three categories: (1) recipient-related factors (parity, body condition, estrus synchronization, corpus luteum characteristics); (2) laboratory factors (cleavage, blastocyst formation, degeneration, embryo grade, developmental stage, cryopreservation); and (3) environmental factors (temperature–humidity index, transport time). Mean comparison and chi-square analyses revealed significant differences in pregnancy rates based on corpus luteum volume, cleavage rates, blastocyst formation rates, degeneration rates, and embryo grade. In binary logistic regression, categorized increases in blastocyst formation rate, degeneration rate, and embryo grade were associated with a 1.45-fold increase, 0.74-fold decrease, and 0.56-fold decrease in pregnancy odds, respectively; no recipient or environmental variables were independent predictors. These findings indicate that developmental competence of IVP embryos is more critical for pregnancy success than recipient or environmental factors, suggesting that optimizing IVP systems to maximize embryo quality is the most effective strategy to improve reproductive efficiency in ET. Full article

A key objective of the dairy industry is to balance genetic progress with reproductive efficiency. Ovum pick-up followed by in vitro embryo production (OPU-IVP) is a pivotal technology for accelerating genetic gain. However, the relationship between follicle size and oocyte developmental competence in high-producing dairy cows under hormonal stimulation remains to be fully elucidated. This study systematically evaluated the effects of follicle diameter ovum pick-up on OPU-IVP outcomes and the underlying follicular fluid (FF) microenvironment. A total of 109 high-yielding Holstein cows were subjected to ovarian stimulation and OPU. Follicles were categorized as small (2.0–5.9 mm), medium (6.0–9.9 mm), or large (10.0–20.0 mm). Oocyte recovery, quality, and developmental competence were assessed. FF was analyzed for hormonal profiles, including anti-Müllerian hormone (AMH), estradiol (E2), follicle-stimulating hormone (FSH), and progesterone (PROG); oxidative stress markers, including malondialdehyde (MDA), glutathione peroxidase (GPx), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC); and untargeted metabolomics (n = 10 per group). Consistently, oocytes from medium follicles exhibited superior developmental competence, achieving the highest maturation (89.93%), cleavage (72.19%), and blastocyst rates (41.88%). In contrast, large follicles had a low recovery rate (32.64%), a high proportion of degenerated oocytes (32.00%), and reduced embryonic efficiency. Metabolomic profiling revealed distinct microenvironmental differences, with medium follicles enriched in pathways like pyruvate metabolism and arachidonic acid metabolism indicating an optimal metabolic state. Hormonally, AMH decreased while E2 and PROG increased with follicle size. Large follicles exhibited significantly elevated MDA levels, indicating oxidative stress, without a concurrent rise in antioxidant capacity. In conclusion, while small follicles provide an abundant source of morphologically good oocytes, medium follicles (6.0–9.9 mm) represent a distinct “window of competence” for OPU-IVP, characterized by a follicular microenvironment most conducive to embryo production. Excessive reliance on large follicle aspiration should be avoided due to signs of over-maturity and oxidative damage. These findings provide a physiological basis for optimizing OPU strategies to enhance IVP efficiency in high-producing dairy cows. Full article

Embryo spots on potato seed enhance the efficiency of doubled haploid screening by facilitating selection. While the spots are known to involve anthocyanin accumulation, their genetic regulation remains unclear. Here, loci and genes regulating spot formation were investigated. An F₁ population was generated by crossing the haploid inducer IVP101 (embryo-spotted male parent) with the diploid inbred line Y8 (non-spotted female parent). Subsequent BSA-seq of the extreme F₁ pools mapped a locus to chromosome 10 (49.96–54.31 Mb). QTL mapping via a high-density genetic map of the F₂ segregating population (derived from F₁ selfing) identified four QTLs (on chromosomes 2, 5, 10, 11). These included the QTLs qSP10-1 (explaining 23.85% of phenotypic variance) and qSP11-1 (18.23%). qSP11-1 overlapped with the reported P locus encoding flavonoid 3′,5′-hydroxylase (F3′5′H), whereas qSP10-1 confirmed the BSA-seq results. Integration of the BSA-seq and QTL mapping results narrowed the target gene locus to a 384.6 kb interval at the end of chromosome 10. Transcriptome sequencing of spotted vs. non-spotted F₁ seed, together with gene expression profiling in the qSP10-1 interval, identified five differentially expressed candidate genes. These findings clarify the genetic basis of potato embryo spot formation and provide a reference for breeding and further research. Full article

VPS13A disease is a rare, autosomal-recessive, neurodegenerative disorder characterized by involuntary movements, orofacial dystonia, seizures, psychiatric symptoms, and the presence of spiky, deformed red blood cells (acanthocytes). The disease is caused by mutations in the VPS13A gene, which encodes the VPS13A protein (previously known as chorein). This protein is a member of the family of bridge-like lipid transport proteins, involved in bulk lipid transfer between membranes and intracellular vesicle trafficking. We describe the case of a 37-year-old woman with gait instability, semi-flexed legs, and involuntary distal muscle movements. Genetic testing was performed using next-generation sequencing (NGS), followed by molecular analysis. Fibroblasts from the patient, her mother, and a healthy control were analyzed by immunofluorescence and Western blotting. NGS identified a novel homozygous 2.8 kb deletion encompassing exons 69–70 (69–70del) of the VPS13A gene (NM_033305.3). The same variant was detected in the patient’s mother in a heterozygous state and her brother in a homozygous state. Although other deletions in the gene have been described, a comprehensive search of population variant databases and the existing literature did not reveal previous reports of this deletion. Fibroblasts from the patient, her mother and a healthy control were characterized. Functional assays showed a complete absence of the VPS13A protein in the patient’s fibroblasts. This study expands the mutational spectrum of VPS13A-linked VPS13A disease and underlines the importance of comprehensive genetic analysis in atypical cases. Full article

The developmental competence of oocytes is a critical limiting factor in bovine in vitro embryo production (IVEP). Our study aimed to investigate the relationship between the intrafollicular concentrations of interleukin-10 (IL-10) and progesterone (P4), follicle characteristics, and the subsequent developmental success of bovine oocytes. Follicular fluid (FF) and corresponding cumulus–oocyte complexes (n = 314) were collected from FSH-stimulated heifers. A novel, high-sensitivity Surface Plasmon Resonance Imaging biosensor was used to quantify IL-10, while P4 was measured by an enzyme-linked fluorescent assay. Oocytes were individually cultured to assess cleavage (Day 3) and blastocyst formation (Day 7). Statistical analysis revealed that intrafollicular IL-10 concentration was a significant positive predictor of developmental success, significantly correlating with blastocyst rate (ρ = 0.29, p = 0.016). Oocytes from follicles with IL-10 concentrations above an optimized cutoff of 142.16 pg/mL had a 16.33-fold greater chance of developing into a blastocyst (p = 0.006). A predictive model combining IL-10 and oocyte morphology demonstrated the highest accuracy for predicting blastocyst success (AUC = 0.724). Conversely, poor oocyte morphology (Grade 4) and large follicular volume (>1200 µL) were significantly associated with developmental failure. Intrafollicular P4 concentration was not directly correlated with embryo development but rather with follicle size. Our findings identify intrafollicular IL-10 as a potent biomarker for predicting bovine oocyte competence and suggest that its quantification using sensitive biosensor technology could enhance the efficiency of IVEP programs. Full article

In this study, we consider and analyze an inhomogeneous Whittaker-type differential equation of the form ${\displaystyle \frac{d^{2}y\left(x\right)}{d{x}^2}}+{\displaystyle \frac{1}{x}}{\displaystyle \frac{dy\left(x\right)}{dx}}-{\alpha }^2\left(x^2-{\beta }^2\right)y\left(x\right)=g\left(x\right)$, where $\alpha$ and $\beta$ are given parameters. We investigate the analytical structure of its solution through the application of the Whittaker integral representation. The analysis encompasses both initial value problems (IVPs) and boundary value problems (BVPs), wherein appropriate conditions are imposed within a unified analytical framework. Furthermore, a systematic methodology is developed for constructing explicit solutions within the framework of Whittaker function theory. This approach not only elucidates the functional behaviour of the solutions but also provides a foundation for extending the analysis to more general classes of second-order linear differential equations. Full article

A high-order single-step implicit method, the Legendre–Darboux Method of order six (LDM6), is introduced for solving both linear and nonlinear initial value problems. Unlike classical Taylor expansions, LDM6 systematically constructs higher-order derivatives via the Darboux formula with Legendre polynomials, yielding a compact scheme of exceptional accuracy and strong stability. To the best of current knowledge, LDM6 is the only single-step method exhibiting spectral-like behavior, achieving near machine-precision global accuracy while retaining efficiency for large step sizes. Comparative experiments on nonlinear cooling problems and the logistic growth model demonstrate that LDM6 surpasses the classical eighth-stage Runge–Kutta method (RK6) in accuracy, stability, and robustness. It attains unprecedented global errors as low as ${10}^{-38}$ and maintains stability for large steps (e.g., $h=10$), whereas RK6 suffers significant error accumulation. These results establish LDM6 as a uniquely efficient, high-fidelity integrator and the first single-step method with spectral-like accuracy, offering a new paradigm for high-precision time integration. Full article

This study introduces a family of implicit four-point block methods for solving first-order initial value problems (IVPs) with oscillatory solutions. In addition to an eighth-order block method, amplification-fitted and phase-fitted implicit block methods are also derived. The methods are implemented in a predictor–corrector framework, where the predictor is a four-point explicit block method constructed with the corresponding properties. A comprehensive stability analysis is carried out to assess the robustness of the proposed approaches. Comparative evaluations with existing methods demonstrate the superior efficiency of the new algorithms. Numerical experiments further confirm that the proposed techniques provide significant improvements over traditional methods, particularly for oscillatory IVPs. Full article