Search Results (183)

Background: Oxidative stress is a critical factor contributing to male infertility, impairing spermatogonial stem cells (SSCs) and disrupting normal spermatogenesis. This study aimed to isolate and characterize human SSCs and to investigate oxidative stress-related gene expression, protein interaction networks, and developmental trajectories involved in SSC function. Methods: SSCs were enriched from human orchiectomy samples using CD49f-based magnetic-activated cell sorting (MACS) and laminin-binding matrix selection. Enriched cultures were assessed through morphological criteria and immunocytochemistry using VASA and SSEA4. Transcriptomic profiling was performed using microarray and single-cell RNA sequencing (scRNA-seq) to identify oxidative stress-related genes. Bioinformatic analyses included STRING-based protein–protein interaction (PPI) networks, FunRich enrichment, weighted gene co-expression network analysis (WGCNA), and predictive modeling using machine learning algorithms. Results: The enriched SSC populations displayed characteristic morphology, positive germline marker expression, and minimal fibroblast contamination. Microarray analysis revealed six significantly upregulated oxidative stress-related genes in SSCs—including CYB5R3 and NDUFA10—and three downregulated genes, such as TXN and SQLE, compared to fibroblasts. PPI and functional enrichment analyses highlighted tightly clustered gene networks involved in mitochondrial function, redox balance, and spermatogenesis. scRNA-seq data further confirmed stage-specific expression of antioxidant genes during spermatogenic differentiation, particularly in late germ cell stages. Among the machine learning models tested, logistic regression demonstrated the highest predictive accuracy for antioxidant gene expression, with an area under the curve (AUC) of 0.741. Protein oxidation was implicated as a major mechanism of oxidative damage, affecting sperm motility, metabolism, and acrosome integrity. Conclusion: This study identifies key oxidative stress-related genes and pathways in human SSCs that may regulate spermatogenesis and impact sperm function. These findings offer potential targets for future functional validation and therapeutic interventions, including antioxidant-based strategies to improve male fertility outcomes. Full article

Background: Perioperative care is an integral part of the procedure of a surgical operation, with strictly defined rules. The need to upgrade and improve some individual long-term processes aims at optimal patient care and the provision of high-level health services. Therefore, preoperative care is drawn up with new data resulting from the evolution of technology to upgrade the procedures that need improvement. According to the international literature, a factor considered to be of major importance is high preoperative anxiety and its effects on the patient’s postoperative course. High preoperative anxiety is postoperatively responsible for prolonged hospital stays, increased postoperative pain, decreased effect of anesthetic agents, increased amounts of analgesics, delayed healing of surgical wounds, and increased risk of infections. The use of Virtual Reality technology appears as a new method of managing preoperative anxiety. Objective: This study investigates the effect and effectiveness of Virtual Reality (VR) technology in managing preoperative anxiety in adult patients. Methods: A literature review was performed on 193 articles, published between 2017 and 2024, sourced from the scientific databases PubMed and Cochrane, as well as the trial registry ClinicalTrials, with a screening and exclusion process to meet the criterion of investigating VR technology’s effectiveness in managing preoperative anxiety in adult patients. This systematic review was conducted under the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA 2020) guidelines. Results: Out of the 193 articles, 29 were selected. All articles examined the efficacy of VR in adult patients (≥18) undergoing various types of surgery. The studies represent a total of 2.354 participants from 15 countries. There are two types of VR applications: distraction therapy and patient education. From the studies, 14 (48%) used the distraction VR intervention, 14 (48%) used the training VR intervention, and 1 (4%) used both VR interventions, using a range of validated anxiety scales such as the STAI, VAS-A, APAIS, and HADS. Among the 29 studies reviewed, 25 (86%) demonstrated statistically significant reductions in preoperative anxiety levels following the implementation of VR interventions. VR technology appears to manage preoperative anxiety effectively. It is a non-invasive and non-pharmacological intervention with minimal side effects. Conclusions: Based on the review, the management of preoperative anxiety with VR technology shows good levels of effectiveness. Further investigation of the efficacy by more studies and randomized controlled trials, with a larger patient population, is recommended to establish and universally apply VR technology in the preoperative care process as an effective method of managing preoperative anxiety. Full article

The image of the vasa domus Dei, repeated on two occasions in the book of the prophet Daniel (Dan 1:2 and 5:4), enables Jerome to formulate an explicit judgement on pagan culture. Drawing extensively on a well-established repertoire, he highlights in one case its positive aspects and in the other its negative ones, and so in CDan 1,1,2b, he underlines the presence, at least in philosophy, of some truths drawn from the doctrine of God; in CDan 2,5,4, he discusses the wicked use the heretics make of the saeculares litterae. Full article

The creation of transgenic chickens holds significant promise for the agricultural and biotechnological sectors, offering potential improvements in disease resistance and production efficiency. The preferred method for generating gene-edited chickens involves the genetic manipulation of primordial germ cells (PGCs), making the identification and isolation of these cells a growing focus of research. PGCs are the precursors to sperm and oocytes, responsible for transmitting genetic material to the next generation. In humans, PGCs are characterized by their large size, round nuclei, and refractive lipids in the cytoplasm, and can be identified using periodic acid–Schiff (PAS) staining and the surface marker stage-specific embryonic antigen 1 (SSEA1). Similarly, chicken PGCs express SSEA1, but their most specific marker is the chicken vasa homologue (CVH), the avian equivalent of the RNA-binding factor gene vasa. However, SSEA1, along with other known surface markers, does not bind to all PGCs or lacks specificity, while CVH, although highly specific to PGCs, is intracellular and unsuitable for isolating viable cells. This study aims to develop an antibody targeting a PGC surface marker with the same specificity as CVH. Despite the importance of identifying surface markers for PGC characterization, to date, such reagents are limited. To address this, whole chicken PGCs were injected into mice, leading to the generation of a panel of monoclonal antibodies. One antibody was found to bind cultured chicken PGCs and showed reduced expression upon differentiation with retinoic acid, indicating its specificity to PGCs. Immunoprecipitation followed by mass spectrometry identified the antigen as myosin heavy chain-like (MYH9) protein. The antibody, αMYH9, was further characterized and shown to bind circulating PGCs and embryonic gonadal PGCs (Hamburger Hamilton (H-H) stage 30, embryonic day 6.5–7). Whilst our primary aim was to determine the binding to PGCs, further investigation is required to determine potential binding to somatic cells. In conclusion, this study provides the characterization of a surface marker for chicken PGCs, with significant implications for advancements in avian genetic preservation, agriculture, and biotechnology. Full article

The cryopreservation of ovarian tissues from fish has recently been carried out for several endangered and commercially valuable species. However, previous studies in this context have focused on the cryopreservation of immature ovaries—mainly through slow freezing and vitrification—which requires specialized freezing equipment or higher cryoprotectant concentrations to keep cell viability. Therefore, the aim of this study was to explore a convenient, rapid, efficient and less toxic method for the cryopreservation of ovaries at the stage of vitellogenesis from yellow drum (Nibea albiflora), an economically important marine fish. The ovaries at the stage of vitellogenesis were isolated and cut into blocks of approximately 1 cm³, then cryopreserved with 15% propylene glycol (PG), fetal bovine serum (FBS) and 0.2 M trehalose as cryoprotectants. Finally, the samples were treated using three different freezing procedures, including a −80 °C refrigerator, liquid nitrogen, and their combination. After 7 days, the tissues were thawed and digested, and the cell survival rates and gene expression levels were detected using cell viability assay kits and qRT-PCR, respectively. The results of the viability assay showed that the procedure of ovarian tissue storage at −80 °C in a refrigerator for 1 h, followed by transfer to liquid nitrogen, resulted in the highest cell survival rate (>90%). Furthermore, the germ cells at various phases were of normal size; presented a full, smooth surface and regular shape; and did not show any signs of cell rupture, atrophy, depression, granulation or cavitation. Furthermore, the qRT-PCR results revealed that genes related to reproductive development, such as vasa, foxl2, zp3 and gsdf, were all down-regulated under the optimal protocol, while the expression of the nanos2 gene (which is specifically distributed in oogonia) maintained a higher level, similar to that in the control group. This indicated that the viability of germ stem cells (oogonia) was not weakened after freezing and that oogonia could be isolated from the cryopreserved ovaries for germ cell transplantation. The present study successfully establishes an optimal cryopreservation protocol for ovarian tissues from Nibea albiflora, providing reference for the preservation of ovaries at the stage of vitellogenesis from other species. Full article

The superfamily Alpheoidea comprises eight families: Alpheidae, Barbouriidae, Bythocarididae, Hippolytidae, Lysmatidae, Merguiidae, Ogyrididae and Thoridae. Alpheoids are characterized by possessing two pairs of chelate pereopods, a multiarticulate carpus on pereopod 2, and a narrow strip as the last article on maxilliped 2. However, during the inspection of the reproductive system (RS) of several alpheoids, we consistently observed the presence of ejaculatory bulbs (EBs) in vasa deferentia (VDs) of these shrimps. To investigate whether the presence of EBs in the RS is a conserved trait among Alpheoidea representatives, we analyzed as many species as possible along the Brazilian coast: Alpheidae—5 genera, 19 spp., Hippolytidae—2 genera, 2 spp., Lysmatidae—2 genera, 10 spp., Merguiidae—1 genus, 1 sp., Ogyrididae—1 genus, 2 spp., and Thoridae—1 genus, 1 sp. In addition, we examined representatives of the superfamilies Atyoidea (1 family, 2 genera, 2 spp.), Nematocarcinoidea (1 family, 1 genus, 2 spp.), Palaemonoidea (2 families, 4 genera, 4 spp.) and Processoidea (1 family, 2 genera, 2 spp.) to determine whether EB are present in these groups. Among the groups analyzed, except for the family Alpheidae, most species of alpheoids exhibit an expansion on the ventral portion of the VD in continuity with the lumen of the vas deferens, i.e., the EB. This structure increases the surface area of the VD, consequently increasing the quantity of the seminal material to be ejaculated onto the female. We did not observe the presence of EB in any other of the analyzed superfamilies, suggesting that this structure is exclusive in Alpheoidea. In conclusion, the presence of EB in VD appears to be an exclusive trait in Alpheoidea, being considered a well-preserved synapomorphic trait in this group, except in the family Alpheidae that do not harbor EB, representing a plesiomorphic condition within this superfamily. Full article

Succenturiate placenta is a rare anatomical variant characterized by one or more accessory lobes connected to the main placental mass by fetal vessels. While frequently asymptomatic, this condition can lead to serious maternal–fetal complications if not diagnosed prenatally. Early detection through advanced ultrasonographic techniques plays a critical role in guiding obstetric management and reducing adverse outcomes. Objective: To describe and analyze the prenatal diagnosis, sonographic characteristics, clinical management, and maternal–fetal outcomes of succenturiate placenta cases diagnosed over a ten-year period at a tertiary care center. Methods: We conducted a retrospective observational study of nine pregnancies diagnosed with succenturiate placenta between 2014 and 2024. Data collected included maternal demographics, ultrasound findings, type of cord insertion, presence of associated anomalies such as velamentous cord insertion or vasa previa, vaginal or cesarean delivery, complications, and neonatal outcomes. Ultrasound evaluation was scored based on a four-point checklist assessing key diagnostic steps. Results: Five of the nine cases (55.6%) presented isolated succenturiate placenta, while four (44.4%) were associated with velamentous cord insertion. No cases of vasa previa were identified. Obstetric outcomes included three vaginal deliveries (33.3%), two instrumental (22.2%), and four cesarean sections (44.4%), one of which was emergent due to fetal distress. Complications occurred in 44.4% of cases, with intrapartum bradycardia being the most common. One neonatal death was reported due to placental abruption. The quality of the ultrasound diagnosis was high in most cases, though transvaginal scanning was inconsistently applied. Conclusions: Prenatal identification of succenturiate placenta via detailed ultrasound, including color Doppler and targeted assessment of cord insertion, is essential to minimize risks associated with this condition. Standardized diagnostic protocols can improve detection rates and enable timely clinical decisions, ultimately improving maternal and neonatal outcomes. Full article

To address the growing consumer demands for improved fish meat quality, desirable morphological traits, and sustainable production practices, researchers have intensified efforts in the selective breeding and genetic improvement of carp (Cyprinus carpio) varieties. However, traditional breeding methods are often time-consuming and inefficient, which poses challenges to the sustainable development of the carp aquaculture industry. The establishment of germ stem cell lines offers a crucial tool for the study of germ cells, genetic improvement, and species conservation. In this study, we successfully established a spermatogonial stem cell line (YRSSCs) from Yellow River carp (Cyprinus carpio haematopterus) that can be cultured in vitro for the long term. We optimized the culture conditions to maintain their self-renewal and differentiation capabilities. The results demonstrated that YRSSCs have a diploid karyotype and can stably proliferate for over a year in L-15 medium supplemented with 5 mmol/L HEPES, 50 μmol/L β-mercaptoethanol, 15% FBS, 2 ng/mL bFGF, 2 ng/mL LIF, 1% carp serum, 800 IU/mL penicillin, 0.8 mg/mL streptomycin, 2 μg/mL amphotericin B, 1% zebrafish embryo extract, and 1% glutamine at 30 °C in the absence of CO₂. The cells exhibited a typical germ stem cell gene expression profile, with strong expression of the vasa, plzf-a, and Oct4-a genes. Additionally, this study found that YRSSCs possess the ability to differentiate in vitro and functionally colonize in vivo within recipient bodies. This research explored the establishment of YRSSCs and their differentiation potential both in vitro and in vivo, providing a novel strategy for the genetic improvement of aquaculture fish species through germ stem cell-based gene editing and transplantation technologies. Full article

Objective: Global utilization of assisted reproductive technology (ART) is increasing; however, it is associated with adverse perinatal outcomes. Placental and umbilical cord abnormalities contribute significantly to these negative outcomes. However, it remains unclear whether ART independently increases the risk of such abnormalities. This study aimed to investigate the association between ART and key umbilico-placental abnormalities, after adjustment for confounders. Methods: In this retrospective cohort study, singleton pregnancies receiving routine antenatal care (January 2015 to June 2024) at the 3rd Department of Obstetrics and Gynecology, Aristotle University of Thessaloniki, Greece, were analyzed. Pregnancies conceived via ART were compared to those conceived spontaneously. To investigate placental and cord anomalies, this study employed multiple logistic regression. This approach adjusted for various confounders, including maternal age, BMI, parity, smoking status, history of previous cesarean section, diabetes mellitus, and thyroid disease. Results: This study included a total of 13,854 singleton pregnancies, of which 647 were conceived via ART. ART was significantly associated with an increased risk of placenta previa (aOR 1.99, 95% CI 1.10–3.61), low-lying placenta (aOR 1.71, 95% CI 1.38–2.11), bilobate placenta (aOR 2.81, 95% CI 1.92–4.11), single umbilical artery (aOR 2.62, 95% CI 1.022–6.715), marginal (aOR 1.63, 95% CI 1.32–2.01) and velamentous cord insertion (aOR 3.13, 95% CI 1.98–4.95), and vasa previa (aOR 5.51, 95% CI 1.28–23.76). Conclusions: ART-conceived pregnancies appear to carry a higher risk for certain placental and umbilical cord abnormalities, potentially contributing to adverse perinatal outcomes. Further studies are required to investigate the pathophysiology underlying these associations. Full article

Background: Fluoroquinolone-induced tendinopathy is a clinically significant adverse effect associated with this class of antibiotics, particularly affecting the Achilles tendon. Despite its growing recognition, the precise pathophysiological mechanisms remain incompletely understood, with hypotheses referencing increased matrix metalloproteinase activity, collagen degradation, and oxidative stress. Methods: This prospective randomized pilot study evaluates the potential protective effectiveness of vitamin E and selenium supplementation in mitigating fluoroquinolone-induced tendinopathy. The study was conducted on 25 patients receiving 500 mg/day levofloxacin antibiotherapy, randomly divided into a control group and an experimental group—vitamin E (400 IU/day) and selenium (200 µg/day), oral supplementation for 28 days. Clinical assessment of the pain level through the VAS score and of functionality through the VISA-A score was performed, alongside ultrasound imaging of the Achilles tendon. To assess potential toxicity and ensure adherence to the supplementation protocol, serial biochemical analyses of serum vitamin E and selenium were performed at predetermined intervals. Results: A significant improvement was observed in pain scores (p = 0.0120) and functional outcomes (p = 0.0340) when comparing the control and experimental groups at the three-month follow-up. Ultrasound analysis revealed reduced tendon thickness and neovascularization, supporting structural recovery. Although the incidence of tendinopathy was lower in the interventional group (13.3% vs. 40%), statistical significance was not reached, possibly due to the small sample size. Conclusions: These findings suggest that antioxidant supplementation with vitamin E and selenium may provide a protective effect against fluoroquinolone-induced tendinopathy, warranting further investigation in larger randomized clinical trials. Full article

Spermatogenesis is a process of self-renewal of spermatogonial stem cells and their proliferation and differentiation to generate mature sperm. This process involves interactions between testicular somatic (mainly Sertoli cells) and spermatogonial cells at their different stages of development. The functionality of Sertoli cells is regulated by hormones and testicular autocrine/paracrine factors. In this study, we investigated the effects of follicle-stimulating hormone (FSH) and testosterone addition on Sertoli cell cultures that undergo hypotonic shock, with a primary focus on Sertoli cell activity. Cells were enzymatically isolated from testicular seminiferous tubules of 7-day-old mice. These cells were cultured in vitro for 3 days. Thereafter, some cultures were treated with hypotonic shock to remove germ cells. After overnight, fresh media without (control; CT) or with FSH, testosterone (Tes), or FSH+T were added to the hypotonic shock-treated or untreated (CT) cultures for 24 h. The morphology of the cultures and the presence of Sertoli cells and germ cells were examined. The expression of growth factors (CSF-1, LIF, SCF, GDNF) or other specific Sertoli cell factors [transferrin, inhibin b, androgen receptor (AR), androgen binding protein (ABP), FSH receptor (FSHR)] was examined by qPCR. Our immunofluorescence staining showed depletion/major reduction in VASA-positive germ cells in Sertoli cell cultures following hypotonic shock (HYP) treatment compared to untreated cultures (WO). Furthermore, the expression of the examined growth factors and other factors was significantly increased in HYP cultures compared to WO (in the CT). However, the addition of hormones significantly decreased the expression levels of the growth factors in HYP cultures compared to WO cultures under the same treatment. In addition, the expression of all other examined Sertoli cell factors significantly changed following HYP treatment compared to WO and following treatment with FSH and or T. However, the expression levels of some factors remained normal following the treatment of Sertoli cell cultures with one or both hormones (transferrin, Fsh-r, Abp, Ar). Thus, our results demonstrate the crucial role of germ cells in the functionality of Sertoli cells and the possible role of FSH and T in maintaining, at least partially, the normal activity of Sertoli cells following germ cell depletion in vitro by hypotonic shock treatment. Full article

Biosensors as advanced analytical tools have found various applications in food safety, healthcare, and environmental monitoring in rapid and specific detection of target analytes in small liquid samples. Up to now, planar electrochemical electrodes have shown the highest potential for biosensor applications due to their simple and compact construction and cost-effectiveness. Although a number of commercially available electrodes, manufactured from various materials on different substrates, can be found on the market, their high costs for single use and low reproducibility persist as major drawbacks. In this study, we present an innovative, cost-effective approach for the rapid fabrication of electrodes that combines lamination of 24-karat gold leaves with low-cost polyvinyl chloride adhesive sheets followed by laser ablation. Laser ablation enables the creation of electrodes with customizable geometries and patterns with microlevel resolutions. The developed electrodes are characterized by cyclic voltammetry and electrochemical impedance spectroscopy, scanning electronic microscopy, and 3D profiling. To demonstrate the manufacturing and biosensing potential, different geometries and shapes of electrodes were realized as the electrochemical transducing platform and applied for the realization of magnetic bead (MB)-labeled biosensors for quantitative detection of food-borne pathogens of Salmonella typhimurium (S. typhimurium) and Listeria monocytogenes (L. monocytogenes). Full article

Metabolic syndrome (MetS) is a subclinical disease, resulting in increased risk of type 2 diabetes (T2D), cardiovascular diseases, cancer, and mortality. Dynamical network biomarkers (DNB) theory has been developed to provide early-warning signals of the disease state during a preclinical stage. To improve the efficiency of DNB analysis for the target genes discovery, the DNB intervention analysis based on the control theory has been proposed. However, its biological validation in a specific disease such as MetS remains unexplored. Herein, we identified eight candidate genes from adipose tissue of MetS model mice at the preclinical stage by the DNB intervention analysis. Using Drosophila, we conducted RNAi-mediated knockdown screening of these candidate genes and identified vasa (also known as DDX4), encoding a DEAD-box RNA helicase, as a fat metabolism-associated gene. Fat body-specific knockdown of vasa abrogated high-fat diet (HFD)-induced enhancement of starvation resistance through up-regulation of triglyceride lipase. We also confirmed that DDX4 expressing adipocytes are increased in HFD-fed mice and high BMI patients using the public datasets. These results prove the potential of the DNB intervention analysis to search the therapeutic targets for diseases at the preclinical stage. Full article

Background/Objectives: Vasa and PL10 belong to the DEAD-box protein family, which plays crucial roles in various cellular functions, such as DNA replication, DNA repair, and RNA processing. Additionally, DEAD-box family genes have also been identified as being related to gonadal development in many species. However, the function of vasa and PL10 in abalone is poorly understood on a molecular level. Methods: In the present study, we individually isolated and characterized the vasa and PL10 orthologs in Haliotis discus hannai (Hdh-vasa and Hdh-PL10). We also characterized the mRNA distributions of vasa and PL10 in various tissues from adult organisms and different embryonic developmental stages using real-time PCR (RT-qPCR) techniques. Furthermore, spatial and temporal expression of Hdh-vasa and Hdh-PL10 throughout embryonic and larval development was examined by whole-mount in situ hybridization (WMISH). Results: The two predicted amino acid sequences contained all of the conserved motifs characterized by the DEAD-box family. Homology and phylogenetic analyses indicate that they belong to the vasa and PL10 subfamilies. We found that vasa and PL10 mRNA were not solely restricted to gonads but were widely expressed in various tissues. WMISH showed that Hdh-vasa and Hdh-PL10 largely overlapped, with both being maternally expressed and specifically localized to the micromere lineage cells during early cleavage stages. By the gastrulation stage, Hdh-vasa were expressed strongly in two bilaterally symmetrical paraxial clusters, but Hdh-PL10 was dispersed in entire endodermal region. Our results suggest that Hdh-vasa-expressing cells are located as a subpopulation of undifferentiated multipotent cells that express Hdh-PL10. As such, we infer that primordial germ cells are specified from these vasa-expressing cells at some point during development, and inductive signals (epigenesis) play an important role in specifying primordial germ cells (PGCs) in H. discus hannai. Conclusions: This study provides valuable insights into the molecular characteristics and expression patterns of Hdh-vasa and Hdh-PL10, contributing to a better understanding of their roles in germ cell specification and early embryonic development in H. discus hannai. Full article

Estrogen receptor β (ERβ) is the most highly expressed subtype in the colon epithelium and mediates the protective effect of estrogen against the development of colon cancer. Indeed, the expression of this receptor is inversely related to colorectal cancer progression. Structurally estrogen-like compounds, including vitamin E components, affect cell growth by binding to ERs. In the present study, cell proliferation was measured by cell counting in a Bürker hemocytometer, and ERβ expression was measured by Real-Time qPCR and immunoenzymatic methods. The results obtained show that natural δ-tocopherol (δ-Toc) and two of its semi-synthetic derivatives, bis-δ-tocopheryl sulfide (δ-Toc)₂S and bis-δ-tocopheryl disulfide (δ-Toc)₂S₂, play an antiproliferative role and upregulate ERβ expression, similar to 17-β-estradiol (17β-E2), in human colon adenocarcinoma HCT8 cells engineered to overexpress ERβ protein (HCT8-β8). These events are not present in HCT8-pSV2neo and in HCT8-β8 pretreated with ICI 182,780, suggesting that they are mediated by the binding of compounds to ERβ, as also boosted by an in silico assay. The antiproliferative effect is independent of the intracellular redox state and (δ-Toc)₂S and (δ-Toc)₂S₂ reduce cell proliferation at concentrations lower than that of δ-Toc and all tested compounds are also able to upregulate ERβ expression. Taken together, the data indicate that, through the involvement of ERβ activity and expression, δ-Toc, (δ-Toc)₂S, and (δ-Toc)₂S₂ may provide potential therapeutic support against colorectal cancer. Full article