Search Results (2,373)

Background/Objectives: The retromer protein complex is involved in various physiological processes, especially endosomal trafficking, and its dysregulation has been linked to Alzheimer’s disease and Parkinson’s disease, as well as VPS35 knockout (KO), causing early embryonic lethality. We aimed to investigate the cellular consequences of VPS35 deficiency. Methods: To investigate the effects of VPS35 loss, we used CRISPR/Cas9 to generate VPS35 KO human embryonic kidney 293 (HEK293) cells. We analyzed changes in retromer component expression, cell proliferation, apoptosis, and mitochondrial dynamics using Western blotting, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and confocal microscopy. Results: VPS35 KO led to a significant reduction in cell proliferation and decreased expression of VPS29 and VPS26, both essential for retromer complex assembly. Consequently, retromer formation was impaired. Compared to control cells, KO cells exhibited elevated levels of cleaved caspase-3, poly(ADP-ribose) polymerase, cytochrome C, and p21, while the expression of Ki-67, CDK4, and cyclin D was reduced. Additionally, VPS35 deletion also promoted mitochondrial fragmentation, associated with increased expression of mitochondrial fission-related proteins. Finally, the rescue experiment using the human VPS35 gene confirmed that the recovery of VPS35 not only led to the recovery of the essential elements constituting the retromer but also the recovery of molecules related to the cell cycle, restoring cell death to a normal level. Conclusions: These findings suggest that VPS35 plays a critical role in cell growth and survival by modulating apoptosis, mitochondrial dynamics, and cell cycle progression. Full article

Background/Objectives: Uveal melanoma (UM) is the most common intraocular malignancy in adults. Although brachytherapy of the primary tumor provides an approximate 80% five-year survival, with time, nearly half of patients experience predominant liver metastases. It was proposed that malignant cells migrate early and stay dormant as they adapt to the liver microenvironment. We propose that cancer vaccine-mediated activation of UM-targeted immunity in primary UM patients could prevent progression of metastatic disease from dormant cells or malignant seeds. Thus, this study explored DNA vaccination as a measure to educate the immune system to recognize the most common UM-associated Q209L tumor driver mutation in GNAQ and GNA11 G-alpha proteins. Methods: Several DNA constructs encoding mutated GNAQ were developed and tested for activation of UM-reactive T cells in HLA-A2/Hd transgenic mice and human T cells ex vivo. Results: Constructs containing immune-enhancing PADRE and VP22-derived epitopes boosted T cell responses against mutant GNAQ, which correlated with reduced experimental lung metastases. Ex vivo dendritic cell-mediated T cell activation with vaccine constructs containing optimized structure produced cytolytic T cells that secreted IFN gamma and killed mutated GNAQ-expressing UM cells in vitro. Conclusions: These findings propose the utility of the fusion DNA vaccines in eliciting T cell immunity against UM cells bearing the Q209L mutation in GNAQ/GNA11 protein to prevent the establishment and progression of metastatic disease. Full article

Enteroviruses (EVs) are small, non-enveloped RNA viruses that can cause diverse clinical outcomes, particularly severe in patients with primary immunodeficiency (PID) due to their impaired ability to clear infections. This study aimed to characterize EV excretion among 138 Tunisian PID patients over a five-year period, to identify circulating EV serotypes and assess their genetic diversity. A total of 558 stool samples were collected and analyzed by virus isolation and intratypic differentiation using RT-qPCR. Molecular typing was performed through Sanger sequencing of the VP1 region and whole genome sequencing using Next-Generation Sequencing (NGS) technologies. Phylogenetic analysis was conducted using the Maximum Likelihood (ML) method. EVs were detected in 55 stool samples from 23 patients. The excretion kinetics of EVs ranged between 30 and 946 days. Thirteen serotypes were identified, including one Poliovirus (PV) and twelve Non-Polio Enteroviruses (NPEVs), predominantly belonging to species B. Two previously unreported serotypes in Tunisia were detected: Coxsackievirus A5 (CVA5) and Echovirus type 19 (E19). In addition, five patients presented enhanced susceptibility to the excretion of successive EV serotypes, and one patient exhibited a co-infection. A possible recombination event was identified in one patient involving Coxsackievirus B5 (CVB5), Coxsackievirus A9 (CVA9) and Coxsackievirus B1 (CVB1) sequences. Phylogenetic analysis showed close genetic relationships with European, American and Asian strains. These findings underscore the dynamic nature of EV circulation and the importance of ongoing molecular surveillance to detect emerging serotypes and guide public health strategies. Full article

A total of 168 fecal samples were obtained from both diarrheic dogs exhibiting clinical symptoms and clinically healthy individuals across four provinces/cities in Northern Vietnam. Based on a polymerase chain reaction method, the Gyrovirus gala 1 (GyVg1) genome was found in 18 (10.71%) of 168 samples. Six representative GyVg1 genomes identified in Vietnam were each 2375 nucleotides long. They shared nucleotide identities of 94.98–98.62%. Whole genome phylogenetic analysis showed that the six obtained GyVg1 strains were grouped into two distinct lineages: GyVg1 I (2/6 strains) and GyVg1 II (4/6 strains). Vietnamese GyVg1 strains were genetically related to Chinese strains. Multiple residue replacements were identified in VP (VP1–VP3) proteins of the Vietnamese GyVg1 strains. A recombination event led to the emergence of the GyVg1/Dog/VNUA-06 recombinant strain. Overall, these observations indicate the presence of GyVg1 viruses in domestic dogs in the North of Vietnam. Full article

During open mitosis, reassembly of the nuclear envelope requires the coordinated recruitment of the ESCRT machinery, initiated by the chromatin-associated factor BAF1 and the nuclear-envelope-associated factor LEM2. Because telomeres are enriched at the reforming envelope, we investigated whether ESCRT factors contribute to telomere integrity. Reduction in the pivotal nuclear ESCRT factor CHMP7 caused DNA damage, heterochromatin disorganization, and telomere defects, including sister telomere associations and telomere free ends. Extending this analysis, we found that additional ESCRT components, including TSG101, VPS28, CHMP4B, and the ESCRT-associated factor AKTIP/Ft1, also contribute to telomere integrity, although with different strengths. Genetic interaction analyses suggest that CHMP7 converges in a common pathway with CHMP4B and AKTIP/Ft1, while it functions in parallel routes to TNKS1, a telomere-specific regulator of the shelterin TRF1. More genetic analyses indicated that BAF1 and LEM2 contribute to safeguarding of telomeres during nuclear envelope reassembly. Because defects in nuclear envelope dynamics and chromatin–membrane coupling are hallmarks of disorders associated with nuclear deformation and fragility, including aging and cancer, our findings contribute a new angle into these conditions and suggest potential targets for selectively modulating telomere maintenance pathways. Full article

Introduction: Accurate nuclear segmentation is essential for the reliable diagnostic interpretation of fluorescence in situ hybridization (FISH) results. However, automated 2D digital algorithms often fail in samples with dense or overlapping nuclei, such as lymphomas, due to the loss of spatial depth information. Here, we tested if AI-based 3D nuclear segmentation can improve the accuracy, reproducibility, and diagnostic reliability of FISH reading in critical situations. Materials and Methods: Formalin-fixed follicular lymphoma sections were FISH-labeled for BCL2 gene rearrangements and digitally scanned in multilayer Z-stacks. The analytic performance in nuclear segmentation of the adaptive thresholding-based FISHQuant, and the freely accessible AI-based NucleAIzer, StarDist, and Cellpose algorithms, were compared to the eye control-based traditional FISH testing, primarily focusing on nuclear segmentation. Results: We revealed that the Cellpose algorithm showed limited sensitivity to low-intensity signals and the adaptive thresholding 2D segmentation, and FISHQuant struggled to resolve densely packed nuclei, occasionally underestimating their counts. In contrast, 3D segmentation across focal planes significantly improved the nuclear separation and signal localization. AI-driven 3D models, especially NucleAIzer and StarDist, showed improved precision, lower variance (VP/VS ≈ 0.96), and improved gene spot correlation (r > 0.82) across multiple focal planes. The similar average number of gene spots per cell nuclei in the AI-based analyses as the eye control counting, despite the elevated number of cell nuclei found with AI, validated the AI nuclear segmentation results. Conclusions: Inaccurate segmentation limits automated diagnostic FISH signal evaluation. Deep learning 3D approaches, particularly NucleAIzer and StarDist, may overcome thresholding and 2D constraints and improve the consistency of nuclear detection, resulting in better classification of pathogenetic gene aberrations with automated workflows in digital pathology. Full article

Although large-scale studies and potential pathways of genes on intramuscular fat (IMF) in livestock have been reported, research on circRNAs in yaks—a unique, low-IMF-content animal species that is native to the Qinghai–Tibetan Plateau—is still lacking. Based on previous high-throughput sequencing results on longissimus dorsi with different IMF content, a novel circRNA encoded by the VPS13C gene (designated as circVPS13C) was found to exhibit significant differential expression. Here, we systematically characterized the function and mechanism of circVPS13C on IMF deposition in yaks by adopting a series of experiments. Sequencing, RNase R processing, and nucleoplasmic separation experiments confirmed the circular structure feature of circVPS13C, and it was predominantly distributed in the cytoplasm. Furthermore, these experiments demonstrated that circVPS13C was mainly distributed in the cytoplasm. The circVPS13C/miR-5606-x/ECHDC3 axis was constructed through ceRNA network analysis and validated by dual-luciferase reporter and rescue experiments. Furthermore, the function of these three potential regulators during IMF deposition was investigated through CCK-8, BODIPY, Oil Red O staining, and qRT-PCR analyses, and results showed that both circVPS13C and miR-5606-x promoted the differentiation and inhibited the proliferation of yak intramuscular preadipocytes, while the function of ECHDC3 was the opposite. In conclusion, circVPS13C could act as a competitive endogenous RNA (ceRNA) sponge to sequester miR-5606-x, thereby relieving the inhibitory effect of miR-5606-x on ECHDC3. Full article

Mouse models for “definitive” polyomavirus nephropathy with lytic viral replication and end-organ disease (PyVN) do not exist. We aimed at defining a PyVN model in Black-Swiss mice (n = 126) by injecting newborn animals with murine polyomavirus (strain A2; 1 × 10⁵ plaque forming units) that led in all mice to a productive intrarenal infection with genetically stable, episomal MuPyV lacking latency. Animals were monitored and morphologic, immunohistochemical, molecular, genetic, and immunological analyses were conducted. Results: Within 3–6 weeks peak PyVN developed characterized by acute tubular injury, lytic replication in up to 14% of tubules (mainly collecting ducts/distal nephrons), high viral gene coverage (up to 3589 viral DNA reads/cell equivalent) and RNA expression (up to 9317 VP1-RNA reads/10⁷ RNA reads), inflammation, DNAemia, and-uria. MuPyV doubling times were high early post-infection (urine: 0.17 day–1.61 day) followed by steady slow viral clearance post day 28 (urine, half-life: 9.90 days). By 54 weeks PyVN had morphologically cleared (no chronic tissue injury) with only qPCR evidence of residual MuPyV in 17% of kidneys/mice. Infection induced an IgM/IgG response (peak plasma IgG titer at 7–30 weeks 1:20,480; low IgG titers in 92% of mice at end of follow-up after one year). During infection, episomal MuPyV remained genetically stable, without significant alterations that could have modified the course of PyVN. The mouse model resembles “definitive” PyVN in humans. It is suited for research on the pathogenesis of PyVN including virally induced tubular injury and immunologic interactions. It facilitates in vivo studies of therapeutic interventions aimed at blocking lytic intrarenalPyV replication. Full article

Background/Objectives: Rabbit hemorrhagic disease (RHD) is an incurable, highly contagious disease caused by different RHD virus (RHDV) strains, such as the coexisting RHDV2 and classical RHDV. Disease control is based on vaccination using different or polyvalent vaccines. This study assessed the protective efficacy, onset of immunity (OOI), and duration of immunity (DOI) of a new recombinant vaccine containing a single active substance developed to target both strains: against RDHV2, highly virulent RDHV2 (hvRHDV2), and classical RDHV strains. Methods: This study included six randomized, controlled, blinded trials in clinically healthy New Zealand White rabbits. Rabbits were grouped to receive the recombinant vaccine or placebo (1:1 ratio) and challenged with RHDV2 (2¹⁵ hemagglutination [HA], infective dose; n = 39 for duration and n = 43 for onset), hvRHDV2 (2¹⁵ HA; n = 48 and n = 40), or classical RHDV (2¹² hemagglutination [HA], infective dose; n = 20 and n = 22) strains to evaluate OOI and DOI. Results: Rabbits receiving the vaccine showed a lower mortality than those receiving placebo upon challenge with any of the three strains. OOI trials showed that vaccinated rabbits exhibited higher levels of antibodies against RHDV2 than controls seven days post-vaccination. DOI trials revealed that, compared with controls, vaccinated rabbits had increased levels of antibodies against RHDV2 across all time points assessed, at least until the day of the viral challenge with any of the RHDV strains (approximately 12 months post-vaccination). Conclusions: This recombinant vaccine is the first to show a durable and robust protection against all tested strains, including the RHDV2, hvRHDV2, and classical RHDV strains, underscoring its potential as a comprehensive tool for RHD prevention. Full article

Type II collagen is a primary fibrillar component of articular cartilage, and its early degradation is a key biomarker of joint-degenerative disorders such as osteoarthritis, rheumatoid arthritis, gout, etc. Reliable detection at low concentrations remains challenging due to limited assay accessibility, complex analytical procedures, and nonspecific responses in multicomponent biological matrices. This research reports the development of a Molecularly Imprinted Polymer (MIP)–based electrochemical sensor engineered for the selective recognition of type II collagen. A series of monomer formulations were evaluated, and the 1AAM:2VP composition produced a well-defined imprinted layer on screen-printed carbon electrodes, yielding the highest electrochemical sensitivity and linearity. The optimized sensor exhibited strong anodic and cathodic responses proportional to increasing collagen concentrations, with a calibration slope corresponding to an R² value of 0.9394. Minimal signal interference was observed, confirming high molecular selectivity. The limit of detection (LOD) was calculated to be approximately 0.065 µg/mL. These characteristics demonstrate that the proposed MIP sensor provides a low-cost, accessible, and highly selective analytical platform suitable for early-stage cartilage degeneration monitoring. Full article

Background/Objective: Infectious bursal disease (IBD) is an acute and highly contagious immunosuppressive disease in chickens caused by infectious bursal disease virus (IBDV). In recent years, a novel variant IBDV (nVarIBDV) has emerged and spread widely, inducing severe immunosuppression and posing a substantial threat to the poultry industry. More importantly, owing to antigenic variations, nVarIBDV can escape the immune protection of the existing vaccines. Therefore, it is imperative to develop a new vaccine that is antigenically matched to nVarIBDV. Methods: The major protective antigen gene VP2 of the representative nVarIBDV strain SHG19 was inserted into the eukaryotic expression plasmid pCAGGS to construct the recombinant plasmid pCASHGVP2. Subsequently, pCASHGVP2 was encapsulated in lipid nanoparticles (LNPs) to form pCASHGVP2-LNP nanoparticles. Finally, using the SPF chicken model, the immune efficacy of pCASHGVP2-LNP was preliminarily assessed by administering two vaccine doses (10 and 20 μg) and two immunization regimens (single or double immunization). Results: Efficient VP2 protein expression from pCASHGVP2 was confirmed by in vitro transfection experiments. The prepared pCASHGVP2-LNP nanoparticles exhibited an optimal particle size distribution and acceptable polydispersity index, indicating a homogeneous formulation. Furthermore, animal experiments showed that the candidate DNA vaccine elicited specific neutralizing antibodies after double immunization and protected immunized chickens from disease induced by nVarIBDV challenge. Conclusions: This study reports the first development of an LNP-encapsulated VP2 DNA vaccine (pCASHGVP2-LNP) against nVarIBDV, highlighting its potential application for the prevention of nVarIBDV. Full article

This study focuses on estimating the nine parameters of the three-diode model (3DM) for photovoltaic (PV) cells by integrating the Atan-Sinc Optimization Algorithm (ASOA) with the Newton–Raphson (NR) method. The ASOA, a population-based metaheuristic approach inspired by the behaviors of the Sech and Tanh functions, systematically generates candidate solutions for the complete set of parameters in the 3DM. For each of these solutions, the NR method is employed to solve the transcendental equation governing the solar cell model, facilitating a precise evaluation of the associated objective function. To guide the parameter estimation process, experimental current-voltage (I-V) and voltage-power (V-P) curves are utilized. The robustness of the proposed methodology is validated through studies on both monocrystalline and polycrystalline solar cells. Computational results reveal that the ASOA effectively navigates the parameter space, while the NR method provides accurate evaluations, resulting in reliable and precise parameter estimations. All numerical validations were conducted using MATLAB software, version 2024b. Full article

Climate change has intensified drought events, compromising popcorn production, particularly in tropical regions. This study aimed to identify popcorn inbred lines with superior water-use efficiency and responsiveness, and to examine the relationships among morpho-agronomic traits associated with expanded popcorn volume per hectare (VP). Fifty inbred lines were evaluated under well-watered (WW) and water-stressed (WS) conditions across two cropping seasons (2020 and 2021). Water deficit was imposed at pre-anthesis, with the permanent wilting point occurring during early reproductive stages in 2020 and during grain filling in 2021. Principal component analysis and efficiency/responsiveness classification were used to characterize line performance. Significant genotype × water condition × season interactions affected all traits. Water stress reduced VP by 75% in 2020 and 46% in 2021, reflecting the differing timing of stress. Line L477 showed high efficiency and responsiveness, while genotypes such as L213, L221, and L222 were inefficient and non-responsive in both years. Under WW, VP was mainly associated with hundred-grain weight, ear length, and grain number per row, whereas under WS, ear diameter and number of rows per ear were the strongest contributors, indicating that the available genetic variability is more effectively exploited through selective morpho-agronomic criteria tailored to each water scenario. Contrasting crosses between efficient and non-responsive lines (L325 and L481) and inefficient but responsive lines (L513, L625, and L689) are recommended to support the development of hybrids that combine high yield under irrigation with resilience under water-stress conditions. Full article

Background/Objectives: Preoperative differentiation between oral squamous cell carcinoma (SCC) and minor salivary gland carcinoma (SGC) remains clinically challenging due to overlapping imaging characteristics. This study aimed to develop a diagnostic model based on quantitative dynamic contrast-enhanced MRI (qDCE-MRI) parameters to distinguish SCC from SGC prior to surgery. Methods: Patients with histopathologic confirmed SCC or minor SGC who underwent preoperative 3.0T qDCE-MRI were recruited. Clinical characteristics and pharmacokinetic parameters, including volume transfer constant (K^trans), reverse reflux rate constant (K_ep), volume fraction of extravascular extracellular space (V_e), plasma volume fraction (V_p), time to peak (TTP), maximum concentration (MAXConc), maximal slope (MAXSlope), and area under the concentration-time curve (AUCt), along with the apparent diffusion coefficient (ADC), were extracted. Univariate and multivariable logistic regression analyses were performed to identify independent discriminators. Diagnostic performance was assessed using receiver operating characteristic analysis, and model comparisons were conducted with the DeLong test. Interobserver agreement was evaluated using intraclass correlation coefficients (ICC). Results: All qDCE-MRI parameters demonstrated excellent interobserver agreement (ICC range, 0.82–0.94). Multivariable analysis identified K_ep (OR = 2620.172, p = 0.001), maximal slope (OR = 1.715, p = 0.024), and tumor location (OR = 5.561, p = 0.027) as independent predictors. The qDCE-MRI model achieved superior diagnostic performance compared with the clinical model (AUC: 0.945 vs. 0.747; p = 0.012). Conclusions: A qDCE-MRI–based model incorporating K_ep and MAXSlope was shown to provide excellent accuracy for preoperative differentiation between oral SCC and minor SGC. Full article

Background: Obesity, defined as an abnormal accumulation of body fat, is becoming a global epidemic. Individuals with obesity may present with increased abdominal fat, which is associated with hypertension, altered respiratory mechanics, higher resting heart rate, and may contribute to an increased cardiovascular risk. Physiological parameters, such as heart rate, blood pressure, respiratory rate, and oxygen saturation, can change hours before the occurrence of a clinically relevant adverse event. Thus, physiological parameters can be considered good predictors of clinical deterioration. Obesity is also associated with physical dysfunctions that can impair physical performance. The non-pharmacological therapeutic strategy for the treatment of obesity involves lifestyle modifications, including a healthy diet and regular physical exercise. Whole-body vibration (WBV) exercise, a type of physical activity, has demonstrated benefits in several specific populations, including obese individuals. Objectives: The objective of this study was to evaluate the immediate effects of a single whole-body vibration (WBV) exercise session, consisting of 15 sets, using a vibration platform (VP) with alternating vertical or lateral displacement, on physiological parameters, perceived exertion, strength, and functionality in obese adults. Methods: Seventy-two obese adult participants were randomly divided into three groups (vertical group, alternating lateral group, and placebo group). Physiological parameters were assessed before, during, and after the intervention, in addition to perceived exertion, functionality, and muscle strength. Results: When comparing the results before and after the intervention, the heart rate–pressure product increased significantly in the alternating lateral group (p = 0.005), and heart rate increased significantly (p = 0.0001) and then decreased significantly (p = 0.030) only in the alternating lateral group. Post hoc analysis revealed a significant increase in perceived exertion in the lateral alternation group, from the period before the intervention to the 10th set (p = 0.006) and from the period before to the period after the intervention (p = 0.011). In the vertical group, a significant increase was observed from the period before the intervention to the 10th set (p = 0.020). Conclusions: In conclusion, considering all the findings of this study, whole-body vibration (WBV) exercise promoted some immediate changes in physiological parameters and perception of effort in obese adults. WBV exercise with the alternating vibration platform induced significant fluctuations in heart rate and increased the heart rate–blood pressure product, although with values within the normal range. Perception of effort increased in all groups. Considering the absence of discrepant changes in physiological parameters, impact on the cardiovascular system, and fatigue, the WBV exercise intervention in side-alternating or vertical vibration vibratory platforms can be considered a viable non-conventional exercise option for the obese population. Full article