Search Results (153)

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

Porcine Sapelovirus (PSV) is widely prevalent in pig herds throughout the world and induces diarrhea, encephalomyelitis, respiratory tract symptoms, and reproductive disorders. However, the epidemiological and genetic evolution characteristics of PSV remain unclear in Yunnan Province. In this study, 1622 fecal samples were collected from pig farms in Yunnan Province. PSV and its co-infection rates with other pathogens were detected; then, the PSV VP1 gene was amplified and sequenced; and the genetic evolution characteristics of the VP1 gene were analyzed. The overall infection rate of PSV in Yunnan Province was 36.50%, and the differences among regions were significant (p < 0.05). The positive rates among different seasons were significantly different (p < 0.01), ranging from 73.33% (autumn) to 19.00% (summer). The PSV positive rate in diarrhea samples (47.26%) was significantly higher (p < 0.001) than that of non-diarrhea samples (31.77%). The co-infection rates of PSV with porcine rotavirus (PoRV) and PSV with porcine epidemic diarrhea virus (PEDV) were 5.07% and 3.04%. A total of 36 VP1 sequences were obtained, and the average identity among the 36 sequences was 85.3%, which was higher than that with other reference strains. Phylogenetic analysis revealed that all 36 PSV strains belonged to the PSV-1 genotype. The VP1 gene was under strong negative selection pressure (average dN/dS = 0.0838); however, the 95th amino acid was under positive selection pressure. Our study revealed the epidemiological, co-infection, and genetic evolution characteristics of PSV in pig herds of Yunnan Province, providing more data for preventing and controlling diarrhea pathogens in pigs. Full article

Potato (Solanum tuberosum L.) is a globally important food crop, but its tetrasomic inheritance and diploid self-incompatibility have limited the discovery of potato genes and progress in breeding. Here, we developed an F₂ segregating population consisting of 174 lines by crossing a self-compatible genome-homozygous diploid line (Y8, female parent) with a heterozygous diploid line (IVP101, male parent), followed by selfing. Using whole-genome resequencing, we constructed a high-density genetic map containing 4464 recombinant bin markers with an average physical distance of 165.51 Kb. Phenotypic evaluation of 8 traits related to yield, tuber shape, and tuber eye number across three environments revealed significant parental differences and wide phenotypic variation within the F₂ population. QTL (Quantitative trait loci) mapping using this genetic map and multi-environment phenotypic data identified 89 QTLs, including 7 previously reported QTLs/genes. In addition, 10 QTLs were stably detected across multiple seasons (stable QTLs). Further genetic effect analysis showed that favorable alleles of these stable QTLs significantly enhanced phenotypic values. Notably, two pleiotropic QTLs were identified on chromosomes 5 and 12; the major-effect QTL on chromosome 12 (qTY-12-6, qTS-12-3, and qTE-12-4) exhibited high phenotypic variance explained (PVE). Its favorable allele from Y8 significantly increased mean tuber weight, tuber number per plant, and promoted rounder tuber shape while reducing eye number, simultaneously improving yield and quality. Collectively, this study provides a reference for genetic mapping using homozygous and heterozygous diploid parents, and the identified QTLs offer valuable genetic resources for potato breeding and molecular mechanism research, enhancing our understanding of the genetic regulation of yield, tuber shape, and eye number in potato. Full article

This study extends the Theory of Functional Connections, previously applied to constraints specified at discrete points, to encompass continuous integral constraints of the form ${\displaystyle {\int }_{x_0}^{x_f}f(x,t)\mathrm{d}x=\mathcal{I}\left(t\right)}$, where $\mathcal{I}\left(t\right)$ can be a constant, a prescribed function, or an unknown function to be estimated through optimization. The framework of continuous integral constraints is developed within the context of initial value problems (IVP) and boundary value problems (BVP). To demonstrate the effectiveness of this analytical approach, examples validate the method and highlight distinctions between satisfying continuous integral constraints via simple interpolation versus functional interpolation. A limitation of the proposed approach is the inability to inherently enforce inequality constraints, such as the positivity constraint $f(x,t)\ge 0$, for modeling probability density functions in classical mechanics. Despite this, numerical experiments on boundary-value problems rarely result in negative values, indicating that the issue occurs infrequently. However, a mitigation strategy based on non-negative least-squares methods combined with Bernstein polynomials is proposed to address these rare cases. This approach is validated through an additional numerical test, demonstrating its efficacy in ensuring nonnegativity when required. Full article

Intravenous push (IVP) administration of controlled substances in hospital settings presents operational challenges related to medication waste, documentation, and diversion risk. This multi-site observational study aimed to quantify the pharmacy workforce time and associated costs linked to IVP waste management across a 16-hospital health system in Southwest Florida. Data were collected from over 4400 controlled substance transactions involving fentanyl, midazolam, hydromorphone, morphine, ketamine, and lorazepam. Methods included automated transaction analysis, manual chart reviews, and software-based compliance case evaluations. Results indicated patterns of partial dose waste, particularly for midazolam (85.2%) and hydromorphone (78.8%), and identified opportunities where documentation efforts could be further optimized through automation. Manual review of 333 incidents required an average of 6 min and 43 s per case, extrapolating to over 496 h of quarterly pharmacy labor or nearly 1985 h annually. Software-based case reviews added another 32 h per quarter or 130 h annually. Additionally, waste receptacle systems incurred over USD 1.1 million in capital costs and USD 322,500 in annual maintenance, with technician labor contributing further operational burden. These findings underscore the resource demands of IVP waste management and support the need for standardized dosing, enhanced documentation workflows, and pharmacy-led interventions to improve efficiency and reduce diversion risk. Full article

Kobuvirus is a new genus of viruses in the Picornaviridae family causing diarrhea in animals. Porcine kobuvirus (PKV) is an important pathogen with a high rate of infection in pig herds. In this study, a total of 10,990 fecal swabs and tissue samples were collected from different areas of Guangxi province in southern China during 2021–2025 and then tested for PKV using RT-qPCR. The results showed a 19.19% (2109/10,990) PKV positivity rate. Sixty-two PKV-positive samples, which were selected according to sampling regions, sampling seasons, and detection Ct values, were used for PCR amplification and gene sequencing. A sequence comparison showed that the nucleotide and amino acid identities of VP1, 2B, and 3D genes were 78.6–99.5% and 83.5–100%, 77.7–99.8% and 80.9–100%, and 90.9–99.8% and 94.9–99.9%, respectively, indicating that the 3D gene was more conserved than the VP1 and 2B genes. The phylogenetic trees based on these three genes revealed that the PKV VP1 gene sequences from different countries could be classified into two groups (Groups I and II), and the PKV VP1 gene sequences obtained from Guangxi province were distributed in Groups I and II and formed independent clades. The 2B and 3D gene sequences could also be classified into two groups (Groups I and II). Bayesian analysis indicated a state of population growth for PKV strains from the time of their discovery until 2009, at which point it began to decline. Amino acid sequence analysis of the VP1 gene identified mutations and insertions in the obtained PKV strains. Recombinant analysis showed that no recombinant event was found in the VP1, 2B, and 3D genes of the obtained strains. The results indicated the geographically specific inheritance and variation in PKV, provided more information on the prevalence and genetic evolution of PKV in Guangxi province, Southern China, and emphasized the importance of regularly monitoring genetic variation in PKV for better comprehension of PKV. Full article

Fiber orientation is an important descriptor of the microstructure for short fiber polymer composite materials where accurate and efficient prediction of the orientation state is crucial when evaluating the bulk thermo-mechanical response of the material. Macroscopic fiber orientation models employ the moment-tensor form in representing the fiber orientation state, and they all require a closure approximation for the higher-order orientation tensors. In addition, various models have more recently been developed to account for rotary diffusion due to fiber-fiber and fiber-matrix interactions which can now more accurately simulate the experimentally observed slow fiber kinematics in polymer composite processing. It is common to use explicit numerical initial value problem-ordinary differential equation (IVP-ODE) solvers such as the 4th- and 5th-order Dormand Prince Runge–Kutta (RK45) method to predict the transient and steady-state fiber orientation response. Here, we propose a computationally efficient method based on the Newton-Raphson (NR) iterative technique for determining steady state orientation tensor values by evaluating exact derivatives of the moment-tensor evolution equation with respect to the independent components of the orientation tensor. We consider various existing macroscopic-fiber orientation models and several closure approximations to ensure the robustness and reliability of the method. The performance and stability of the approach for obtaining physical solutions in various homogeneous flow fields is demonstrated through several examples. Validation of our orientation tensor exact derivatives is performed by benchmarking with results of finite difference techniques. Overall, our results show that the proposed NR method accurately predicts the steady state orientation for all tensor models, closure approximations and flow types considered in this paper and was relatively faster compared to the RK45 method. The NR convergence and stability behavior was seen to be sensitive to the initial orientation tensor guess value, the fiber orientation tensor model type and complexity, the flow type and extension to shear rate ratio. Full article