Search Results (358)

Background: Men who have sex with men (MSM) in Brazil remain disproportionately affected by HIV. Combination prevention strategies, including Pre-Exposure Prophylaxis (PrEP), are critical, yet adherence remains a challenge. This study aimed to identify factors associated with the simultaneous practice of condomless anal sex and non-use of PrEP among Brazilian MSM. Methods: A national cross-sectional study was conducted in 2020 via an online questionnaire disseminated on social media and dating apps. The outcome was defined as reporting condomless anal sex and no PrEP use in the previous year. Bivariate and multivariate logistic regression analyses were performed. Results: Among 1357 MSM participants, a high proportion (69.4%) reported condomless anal sex without PrEP use. Factors significantly associated with this behavior included being younger (18–28 years; AOR: 2.59), identifying as homosexual (AOR: 6.04), bisexual (AOR: 5.30), or pansexual (AOR: 8.67), having a steady partner (AOR: 4.57), engaging primarily in receptive or insertive anal sex, and having a prior STI diagnosis (AOR: 1.49). Conclusions: The confluence of condomless sex and PrEP non-use reveals a significant vulnerability profile among young MSM in Brazil, even within steady relationships. These findings highlight the originality of examining this combined behavioral outcome and underscore the urgent need for targeted, culturally sensitive prevention strategies that address risk perception and enhance PrEP uptake to meet the UNAIDS 2030 goals. Full article

Sweet potato (Ipomoea batatas) is a globally important crop and one of a growing number of plants recognized as naturally transgenic, harboring Agrobacterium-derived T-DNA genes whose functions remain largely uncharacterized. In this proof-of-concept study, we applied CRISPR/Cas9 technology to generate targeted knockouts of the Ib-rolB/C and Ib-rolD-like genes located within the sweet potato cellular T-DNA2 (IbT-DNA2) region. Mutations were introduced into sweet potato callus cultures using an optimized genome editing protocol, with most edits consisting of single-nucleotide insertions. Knockout of Ib-rolB/C did not affect callus growth but significantly reduced levels of chlorogenic acid derivatives. Validation in planta using transient expression in I. batatas leaves confirmed the suppressive effect of Ib-rolB/C disruption on polyphenol content. In contrast, Ib-rolD-like knockout lines showed reduced biomass accumulation and downregulation of cell cycle–related genes, but did not display significant changes in metabolite content in either callus cultures or leaf tissues. These findings suggest that Ib-rolB/C and Ib-rolD-like may differentially contribute to growth and secondary metabolism in sweet potato. Full article

Background/Objectives: Pelvic organ prolapse (POP) is a common condition affecting women. When conservative treatment fails, surgical correction is indicated. Anterior colporrhaphy (AC) is a standard procedure for anterior vaginal wall prolapse repair. Postoperatively, an indwelling urinary catheter (IUC) is typically inserted to prevent urinary retention; however, prolonged catheterization is a known risk factor for urinary tract infection (UTI). This study aimed to evaluate whether postoperative catheterization can be safely shortened from 4 days to 24 h after vaginal POP surgery, and to compare the incidence of urinary retention and UTI between the two groups. Methods: A prospective randomized controlled trial was conducted, including 119 patients scheduled for AC for POP repair. All patients received an IUC after surgery and were randomized to catheter removal after either 24 h (group 1) or 4 days (group 2). Urinary retention was defined as a postvoid residual volume > 200 mL after IUC removal. UTI was diagnosed based on typical symptoms and a positive urine culture (≥10⁵ CFU/mL). Results: Data from 80 patients were analyzed. There were no statistically significant differences in catheter reinsertion rates (15% in group 1 vs. 7.5% in group 2, p = 0.288). The incidence of urinary retention was not influenced by the use of Kelly sutures, concomitant procedures, or patient age. No UTIs were confirmed in either group. Median hospital stay was significantly shorter in group 1 (3 [2–4] days vs. 4 [4–4] days, p < 0.001). Conclusions: Short-term catheterization following anterior colporrhaphy is not associated with increased risk of urinary retention or infection. Reducing catheterization duration results in a shorter hospital stay, which may lower healthcare costs and improve patient throughput. Full article

Background: Emergency contraception (EC), also known as postcoital contraception, is a method used to prevent an unintended pregnancy following unprotected or inadequately protected sexual intercourse. The available options include emergency contraceptive pills or the insertion of an intrauterine device (IUD). Emergency contraception pills contain either levonorgestrel (a single 1.5 mg dose, effective within 72 h) or ulipristal acetate (a single 30 mg dose, effective within 120 h), both of which are most effective when taken as soon as possible after unprotected intercourse. Another highly effective option is the insertion of a copper or levonorgestrel-releasing intrauterine device, although IUDs are not registered for EC use in all countries. The aims of this cross-sectional, comparative, observational study were to collect data on the emergency contraception methods used by adolescent girls and young women to examine their association with various factors, such as religious beliefs, and to evaluate the effectiveness of different emergency contraception methods, including hormonal options and intrauterine devices. Methods: Data were collected from 240 women who attended our Family Planning Clinic using a structured questionnaire that included items on their demographic characteristics, religious beliefs, medical history, lifestyle factors, contraceptive use and side effects, prior use of emergency contraception, method selected, and reasons for seeking emergency contraception. Descriptive statistics were used to summarize the data, comparisons between religious groups were conducted using chi-square tests, and factors related to the timing of emergency contraceptive use were investigated using multinomial logistic regression analysis. Results: Most of the reasons for emergency contraception use did not differ significantly between Christian and Muslim participants. However, Christians were significantly more likely to use emergency contraception due to missed contraceptive doses (20.9% vs. 6.7%, p = 0.004) or the failure to take a progesterone-only pill (19.1% vs. 3.3%, p = 0.001). Levonorgestrel was the most frequently used method in both groups (48.9% of Christians vs. 60% of Muslims, p = 0.132), followed by ulipristal acetate (30.9% vs. 40%, p = 0.180). Notably, 18.5% of Christian participants used an intrauterine device (IUD) for emergency contraception, while no Muslim participants reported IUD use (p < 0.001), indicating a significant difference potentially influenced by cultural or religious factors. Conclusions: Both religious and individual sociodemographic factors affect not only the choice of emergency contraception but also the urgency with which the emergency contraception is used. Interventions aimed at improving contraception education, addressing partner-related challenges, and promoting timely access could improve reproductive health outcomes. Full article

Background: Complicated pneumonia (CP) in children presents in various forms—including empyema, necrotizing pneumonia (NP), necrotizing pneumonia with pleural effusion (NP + PE), and parapneumonic pleural effusion (PPE)—and is associated with significant morbidity despite advances in antimicrobial therapy. This study aimed to describe and compare the clinical characteristics, laboratory findings, antibiotic use, and outcomes across different CP subtypes in hospitalized children and to assess the impact of prior antibiotic use on presentation and treatment outcomes. Methods: This retrospective observational study included 58 children admitted with CP to tertiary hospitals in Jordan. Patients were categorized into four subtypes: empyema (n = 4), NP (n = 4), NP + PE (n = 17), and PPE (n = 33). Demographic data, clinical features, laboratory results, antibiotic regimens, and clinical outcomes were analyzed. Multivariable regression was used to identify predictors of prior antibiotic use. Results: Fever and cough were the most common symptoms (96.6%). Over 40% of patients had received antibiotics prior to admission. Those pre-treated had significantly longer symptom duration (8.2 vs. 4.5 days, p < 0.001), longer hospitalization (18.2 vs. 14.6 days, p = 0.023), and more frequent chest tube insertion (66.7% vs. 35.3%, p = 0.019). Streptococcus pneumoniae was the most common organism isolated in culture-positive cases. Vancomycin-based regimens were the most frequently used treatments. Univariate regression analysis showed that patients with prior antibiotic use had significantly higher odds of longer hospitalization duration (OR = 1.11, p = 0.028) and chest tube insertion (OR = 3.67, p = 0.021). Conclusions: Complicated pneumonia in children remains a diverse and clinically significant condition. The findings demonstrate that prolonged symptom duration prior to hospitalization and certain clinical interventions were associated with prior antibiotic exposure. These results provide insight into local disease patterns and prescribing behaviors, which may help inform strategies to optimize antimicrobial stewardship and improve care pathways for affected children. Full article

Serotonin (5-HT) is a vital intercellular messenger with diverse signaling functions throughout the human body. We have characterized and implemented a novel, in vitro fluorescence-based method of measuring 5-HT binding to gain a fuller understanding of the interactions between 5-HT and its receptors. This method involves expression of 5-HT receptor proteins in cultured cells with the fluorescent, non-canonical amino acid l-3-(6-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid (ANAP) incorporated into the ligand binding site. ANAP fluorescence was quenched in solution by both 5-HT and dopamine. Time-resolved photoluminescence and transient absorption spectroscopy confirmed that ANAP quenching by 5-HT occurs via a charge-transfer process that recovers through back-electron transfer on the nanosecond timescale. Supported by density functional theory calculations, this process likely involved an ANAP reduction by 5-HT. To test this method on intact receptors in a cellular context, we expressed 5-HT_3A receptors (5-HT-gated ion channels) in HEK293T cells with ANAP inserted co-translationally into the transmitter binding site. Fluorescently labeled 5-HT_3A receptors were functional and activated by 5-HT, as assessed by whole-cell patch clamp. Addition of 5-HT caused a concentration-dependent quenching of fluorescence from ANAP-tagged channels in intact cells and unroofed plasma membranes, demonstrating the utility of this method for measuring 5-HT binding to its receptors. Collectively, these results delineate a technique for measuring transmitter binding that can be widely adopted to explore 5-HT binding not only to 5-HT₃ receptors, but to any 5-HT receptor, transporter, or binding protein in heterologous expression systems. Full article

Background/Objectives: To investigate the long-term changes in conjunctival bacterial flora before and after dacryoendoscopic recanalization for lacrimal passage obstruction using silicone tube intubation. Methods: This prospective study included 135 eyes with lacrimal passage obstruction that underwent lacrimal passage recanalization and were followed for at least one year. The silicone tubes inserted during surgery were removed three months postoperatively in all cases. The study period was from November 2018 to January 2025. Conjunctival samples were obtained before surgery and at 12 months postoperatively. Aerobic cultures were performed to detect bacterial flora. The culture positivity rate, number of bacterial species identified, and proportion of commensal bacteria were compared before and after surgery. Results: The bacterial culture positivity rate significantly decreased from 36.3% preoperatively to 20.0% postoperatively (p = 0.003). The number of bacterial species detected decreased from 15 to 6, with Gram-negative bacilli decreasing from 6 species to 1. In contrast, the proportion of commensal bacteria such as coagulase-negative staphylococci and Corynebacterium spp. relatively increased from 49.1% to 80.7%. No drug-resistant bacteria were detected postoperatively. Conclusions: Dacryoendoscopic recanalization for lacrimal passage obstruction was shown to achieve long-term normalization of the conjunctival bacterial flora by reducing pathogenic and drug-resistant bacteria and increasing commensal bacteria. These findings suggest that the procedure prior to intraocular surgery in patients with lacrimal obstruction may reduce the risk of postoperative infection. Full article

SARS-CoV-2 persists worldwide, driving the demand for effective antivirals that inhibit replication and limit the emergence of resistant variants. Lycorine, a non-nucleoside inhibitor of SARS-CoV-2 RNA-dependent RNA polymerase, exhibits antiviral activity without direct mutagenic effects. Here, we examine the occurrence of single-nucleotide variants (SNVs) and insertions/deletions (indels) in SARS-CoV-2 B.1.499 strain during serial passages in Vero cells, comparing lycorine-treated cultures (2.5 and 5 µg/mL) with untreated controls. Whole-genome sequencing was used to assess mutation patterns and frequencies. Lycorine-treated passages displayed greater variant diversity than controls, with fixed mutations mainly affecting non-structural proteins (Nsp3-F1375A, Nsp5-L50F, and Nsp14-G265D) and the envelope protein (E-S6L). A 15-nucleotide deletion in the spike gene (QTQTN motif) occurred in both groups but became fixed only in untreated passages, suggesting negative selection under lycorine pressure. Notably, the L50F mutation in Nsp5, previously linked to nirmatrelvir resistance, was found exclusively in lycorine-treated passages. Additionally, a 1-nucleotide deletion in the accessory gene ORF8, detected only under lycorine treatment, resulted in a frameshift mutation that added four amino acids, potentially altering the protein’s function. Overall, lycorine induces a distinct mutation profile, favoring replication-related variants while suppressing deleterious deletions. These findings suggest potential mechanisms of cross-resistance and highlight the importance of monitoring resistance during clinical use. Full article

Rational design of capripoxvirus-based vaccine vectors can be achieved by knockout of immunomodulatory genes. In this study, the effect of knockout of the immunomodulatory genes LSDV005, LSDV008 and LSDV066 on the replication of Lumpy skin disease virus in cell cultures and the immune response to an integrated foreign antigen were assessed. The knockout of genes was performed by homologous recombination under conditions of temporary dominant selection. It was found that single knockout of the LSDV005 gene and the LSDV008 gene did not affect the replicative activity of recombinant viruses in vitro (Atyrau-5 and Atyrau-B). Both single knockout of the LSDV066 gene and in combination with knockout of LSDV005 or LSDV008 led to a decrease in the replicative activity of recombinant LSDVs. The recombinant Atyrau-5J(IL18) with LSDV005 gene knockout induced production of antibodies to the integrated antigen in mice. Prime-boost vaccination with all studied recombinants increased the level of interferon-γ. In addition, during immunization with the recombinant Atyrau-5J(IL18) secretion of interleukin-2 was significantly increased. The study of the functions of immunomodulatory genes and their effect on the expression of inserted sequences of foreign antigens is promising for the creation of highly effective polyvalent vector vaccines for animals. Full article

Background: Digital polymerase chain reaction (dPCR) is a highly sensitive molecular method that allows rapid detection of bacterial DNA and resistance genes, requiring only a small blood volume. Although not a new technology, its application in pediatric patients with suspected catheter-related bloodstream infection (CRBSI) remains limited. Case presentation: A 16-year-old female, diagnosed with recurrent acute myelogenous leukemia, received re-induction chemotherapy through a peripherally inserted central venous catheter (PICC). The patient developed a fever, and the blood culture (BC) drawn from the PICC was positive for methicillin-resistant S. epidermidis, leading to suspicion of CRBSI. Several antibiotics were used, and the PICC was replaced. Eventually, the fever subsided, and the BC was negative after PICC removal. The levels of S. epidermidis-specific DNA sequences and mecA genes were correlated with the results of the BC and clinical course. Turnaround time was significantly shorter in dPCR (3.5 h) than in the BC (14–21 h); dPCR was performed using only 400 µL of blood. Conclusions: This case highlights the potential of dPCR as a complementary tool to conventional BCs in the management of pediatric CRBSI. dPCR may support rapid decision-making and monitoring of the treatment response, particularly when sample volumes are limited. Full article

In traditional Chinese timber architecture, “column reduction” (Jian Zhu Zao) and “column relocation” (Yi Zhu Zao) enhances spatial continuity, yet often produces bending-dominated, material-intensive frames. This study develops a generative frame system that encodes raised beam logic into a parametric line-model workflow and couples it with simulation-based optimization. Informed by case analysis, the tool implements three lateral strategies—ridge-support revision, insertion of inclined members, and inclination of originally horizontal members—and one longitudinal strategy—longitudinal truss formation—whose use is governed by a user-defined historical authenticity parameter. Structural responses were evaluated using Karamba3D, and cross-section sizing was searched using Wallacei under gravity-dominant loading. The results indicate clearer load paths, greater axial-force participation, and reduced bending, yielding lower maximum displacements at comparable self-weight; moreover, the performance ranking aligns with the calibrated authenticity loss schedule, suggesting that the authenticity controller also acts as a practical proxy for expected stiffness gains. The framework improves design and modeling efficiency while offering quantitative decision support for culturally sensitive conservation and imitation design. Limitations include line-model idealization, simplified timber and joint behavior, gravity-only loading, and a modest historical corpus. The approach is extensible to other traditional systems via parameter and rule adaptation. Full article

Architectural education programs are rapidly expanding the use of immersive technologies worldwide. An increasing number of architecture schools have incorporated 360-degree videos as one of the accessible and cost-effective immersive tools. Despite their availability and ease of use, research on their effectiveness as a learning tool in architectural pedagogy remains limited and mostly focused on architectural design education. Few studies have discussed their application in theoretical courses and their potential to support cognitive understanding of architecture. Learning cultural heritage is considered a foundation of architectural theory. This study examines how the utilization of 360-degree videos, compared to conventional 2D videos, supports spatial cognition, learning performance and experience in cultural heritage education among undergraduate architecture students. An educational experiment was conducted with 89 students in their second year of the architecture degree at the Applied Science Private University, Jordan. Both 360-degree videos and conventional 2D videos were inserted as learning tools within the curriculum of History of Architecture 1 and 2 courses. A mixed-research-method framework, including observation and a post-test survey, was carried out. Using SPSS and Excel programs, the data were analyzed through a set of statistical analyses such as paired-sample t-tests, AHP, and basic descriptive analysis. The findings demonstrate that students were highly immersed and motivated when using 360-degree videos. Compared to conventional 2D videos, 360-degree videos enhanced students’ spatial cognition, performance, engagement, and participation levels in both face-to-face and online courses. These results suggest that 360-degree videos can serve as a sufficient, low-cost, and equipment-free learning tool, responding to the urgent need to utilize technologies in both theoretical and practical architectural pedagogy. Full article

Sex determination is a fundamental biological process governing animal reproduction. Although substantial progress has been made in elucidating its genetic basis, the genetic architecture underlying complex sex determination systems remains poorly understood. In this study, we identify sex-associated insertion–deletion (indel) variants, screen candidate genes, and compare sex-associated variation across populations with different genetic backgrounds in the Fujian oyster (Magallana angulata). Based on whole-genome resequencing data of a culture strain (designated FL), a total of 299,774 high-quality indels were identified. By integrating genome-wide association analysis (GWAS), fixation index (F_ST) analysis, and sex-biased genotype frequency comparisons, 77 overlapping sex-associated indels were identified, predominantly clustered within a 1.8 Mb (8.3–10.1 Mb) region on chromosome 9. Principal component analysis (PCA) based on the sex-associated markers and their subsets consistently separated male and female individuals in the FL strain. For two representative sex-associated indels, PCR-based genotyping methods were developed and validated. Functional annotation identified putative candidate genes for sex determination, including PKD1L1, 5-HTRL, SCP, and CCKRa. Comparative analysis of variants within PKD1L1 across wild, farmed, and selectively bred populations revealed a progressive enrichment of male-linked alleles in domesticated and selectively bred groups, particularly in male individuals. This study provides direct evidence that sex in the Fujian oyster is genetically determined and reveals that domestication and artificial selection may drive the emergence of major sex-determining loci, offering important insights into the genetic basis of sex determination in the Fujian oyster, and establishing a theoretical and practical foundation for molecular marker-assisted breeding of monosex lines for this species. Full article

The retinal pigment epithelium (RPE) is strongly involved in the pathogenesis of several retinal diseases, such as age-related macular degeneration (AMD). RPE models addressing specific pathological pathways are of high importance for understanding cellular pathomechanisms and pre-clinical screening of potential new therapeutics. The goal of this study is to establish standard operation protocols for single-eye porcine RPE preparation for AMD-relevant models of oxidative stress (RPE-Ox) and inflammation (RPE-Inf). Porcine primary RPE were prepared from one eye and seeded into one well of 12-well plates or, for polar differentiation, in transwell inserts. Different coatings (Poly-ᴅ-Lysine and laminin) and serum content of media (10%, 5%, and 1%) were tested to determine optimal culture parameters. For RPE-Ox, cells were treated with NaIO₃, CoCl₂, or erastin; cell viability (thiazolyl blue tetrazolium bromide, MTT), and gene expression (RT-qPCR) were determined. For RPE-Inf, cells were treated with lipopolysaccharide (LPS), polyinosinic/polycytidylic acid (Poly I:C), or tumor necrosis factor alpha (TNF-α); cell viability (MTT), cytokine secretion (ELISA), and gene expression (RT-qPCR) were determined. For transwell plates in RPE-Inf, cell viability (MTT), polar cytokine secretion (ELISA), gene expression (RT-qPCR), and transepithelial electrical resistance (TEER) for barrier assessment were conducted. For RPE-Ox, effective LD₅₀ could be achieved by using 24 h stimulation with 25 µm erastin, seven days after preparation in 5% serum cultures, without coating. For gene expression assessment, the use of Poly-ᴅ-Lysine is recommended. For RPE-Inf, three days of LPS stimulation (1 µg/mL) showed effective cytokine activation with 5% serum on uncoated 12-well plates. Transwell plates are not recommended for cytokine secretion assessment. It can be used for cell barrier assays in which LPS also showed effective cell barrier decrease and gene expression assays. Two specific best practice protocols for the use of porcine single-eye cultures in AMD research concerning oxidative stress and inflammation with optimized parameters were established and are provided. Full article

With the increasing severity of forest pest problems, breeding insect-resistant varieties has become a crucial task for the sustainable development of forestry. The highly insect-resistant triploid Populus line Pb29, genetically modified with BtCry1Ac, served as the maternal parent in controlled hybridization with three paternal Populus cultivars. Hybrid progenies were obtained through embryo rescue and tissue culture. Results showed that 4 °C storage was favorable for pollen preservation, with 84K poplar exhibiting superior pollen viability and embryo germination rates. All progenies displayed significantly lower seedling height and ground diameter growth than the maternal parent (p < 0.05), with some showing leaf shape and branching variations. Among the three crosses, the 84K-sired progeny exhibited the best growth performance but the highest variability. PCR analysis confirmed stable inheritance of the BtCry1Ac and Kan genes from Pb29, showing tight linkage. Progenies carrying BtCry1Ac exhibited detectable gene transcription and toxic protein accumulation, though expression levels varied due to copy number, insertion sites, and potential co-suppression effects. Ploidy analysis suggested all hybrids were aneuploid, with lower survival rates than the maternal parent. Insect-feeding assays confirmed high resistance in all BtCry1Ac-inheriting progenies, with an average larval mortality rate of 97.03%. Mortality rates and death indices significantly correlated with transcript abundance and toxin protein levels. These results demonstrate that BtCry1Ac insect resistance is stably inherited through hybridization. Transgene expression appears co-modulated by copy number, insertion sites, and ploidy status. Simultaneously, it was found that the aneuploid progeny derived from triploid hybridization exhibited growth disadvantages. This provides an important basis for subsequent poplar improvement breeding. Full article