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Blackgram (Vigna mungo (L.) Hepper) is an important legume crop extensively grown in Asia. It is one of the most abundant sources of protein, minerals, and vitamins. Studying genetic variability is an important task for genetic improvement of this crop. Ten blackgram genotypes were experimented on a randomized complete block design with three replications at Sundarbazar, Lamjung during July- October of 2019 to analyze their genetic variability. Plant height at maturity, days to 50% flowering, number of primary branches, branch length, number of pods bearing peduncles, the longest peduncle length, pod length, number of seeds per pod, 1000 seed weight, and grain yield per plant were significantly different between genotypes. BLG0076-2 had produced the highest grain yield per plant (6.19 g), followed by BLG0066-1-1 (5.42 g). The number of pods per plant had the highest genotypic coefficient of variation (GCV) (37.30 %) and phenotypic coefficient of variation (PCV) (46.44 %), followed by the number of pods bearing peduncles (GCV: 34.47, PCV: 46.44). Heritability values for branch length (0.94) are the greatest, followed by seeds per pod (0.92). Genetic advance (GA) was the highest in branch length (20.68), followed by the number of pods per plant (19.79). Similarly, the number of pod bearing peduncles had the highest genetic advance as a percent of the mean (GAM) (66.10), followed by branch length (63.59). Cluster analysis revealed that cluster 2 and cluster 3 had the greatest distance between cluster centroids (24.99), indicating genetic dissimilarity. This study suggests that blackgram genotypes, namely BLG0076-2 and BLG0066-1-1 were potential germplasm for varietal improvement programs.

We investigate theoretically the origins of observed variations in the critical currents of Nb/Al-AlO x /Nb Josephson junctions in terms of various types of disorder. We consider the following disorder sources: vacancies within the Al layer, thickness variations in the AlO x layer, and "pinholes" (i.e., point contacts) within the AlO x layer. The calculations are all performed by solving the microscopic Bogoliubov–de Gennes Hamiltonian self-consistently. It is found that a small concentration of vacancies within the Al layer is sufficient to suppress the critical current, while the presence of a small number of thick regions of the oxide layer induces a similar effect as well. The pinhole scenario is found to result in anomalous behavior that resembles neither that of a pure tunnel junction nor that of a superconductor–normal–superconductor junction but a regime that interpolates between these two limits. We comment on the degree to which each of the three scenarios describes the actual situation present in these junctions. [ABSTRACT FROM AUTHOR]

Introduction: Cardiopulmonary resuscitation (CPR) is an evidence-based intervention that saves lives. In low- and middle-income countries like Nepal, the occurrence of the problem and its outcome are seldom studied. The study aims to highlight the prevalence, performance, and outcome of CPR in government hospitals of Nepal., Methods: A mixed method study was done for two months in central and provincial government hospitals of Nepal. A total of 80 resuscitations were evaluated using a questionnaire based on the American Heart Association 2020 guidelines for cardiopulmonary resuscitation. An in-depth interview was conducted with 15 active participants of the resuscitation in different sites. Thematic analysis was done using the framework of the chain of survival of arrest victims., Results: The overall prevalence of CPR was found to be 1.92% [95% CI: 0.01,0.02] with 5.4% in central hospitals and 0.65% in provincial hospitals with 60% cardiac arrests occurring in the intensive care unit. Estimated time from recognition of the arrest to initiating CPR was 1.9 ±1.4 minutes. Asystole 66.25% was the commonest arrest rhythm and 21.25% had difficulty interpreting rhythm. Only 11.25% of the victims had return of spontaneous circulation and were subsequently transferred for post-arrest care. The qualitative analysis highlighted the lack of trained staff, a dedicated system, feedback mechanism, and provision of post-arrest care., Conclusion: Across various level of Nepal's healthcare system, cardiopulmonary resuscitation is prevalent with poor performance and outcome. To improve outcomes, it is essential to implement standardized procedures and ensure high quality resuscitation delivery before and after the event supported by well-trained healthcare personnel and adequate infrastructure., Competing Interests: The authors have declared that no competing interest exist., (Copyright: © 2025 Hamal et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Fournier's gangrene, a polymicrobial infection affecting the scrotum and perineal area, predominantly affects elderly males, presenting challenges in diagnosis and management. This report examines two cases, with a focus on the rare outcome of bilateral orchidectomy., Case Presentation: A 69-year-old diabetic male presented with severe penile and scrotal pain, leading to the diagnosis of Fournier's gangrene. In another case, a 91-year-old male with a history of Benign Enlargement of the Prostate experienced scrotal pain and ulcerative lesions, resulting in a rapid progression of gangrene. The surgical approaches encompassed multiple debridement and bilateral orchidectomy, underscoring the intricate nature of managing Fournier's gangrene., Discussion: Fournier's gangrene typically originates as a polymicrobial infection in the genitourinary or perianal regions, with common risk factors including diabetes, immunosuppression, and advanced age. Diagnosis often relies on clinical assessment, though imaging aids in complex cases. Early intervention with broad-spectrum antibiotics and repeated debridement remains the cornerstone of treatment., Conclusion: Fournier's gangrene, a rare condition, poses diverse clinical presentations, management strategies, and outcomes. Early recognition, aggressive surgical debridement, and a multidisciplinary approach are essential in managing this condition., Competing Interests: Declaration of competing interest The authors report no conflicts of interest., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Due to the increased prevalence of plastic pollution globally, atmospheric deposition of microplastics (MPs) is a significant issue that needs to be better understood to identify potential consequences for human health. This study is the first to quantify and characterize atmospheric MP deposition in the Eastern United States. Passive sampling was conducted at two locations within the Eastern United States, specifically in remote South Central Appalachia, from March to September 2023. Each location had five sampling periods, with collections over a 21 day period. Samples were processed to remove biological material, and the presence of MPs was confirmed using Raman spectroscopy to match particles based on polymer similarity. The relative average atmospheric MP deposition in South Central Appalachia was determined to be 68 MPs m -2 d -1 . Most verified MPs were fibers, and the most abundant polymer type identified was poly(ethylene terephthalate) PETE. This study's average MP deposition rate is qualitatively comparable to rates reported in other studies that employed a similar methodology in a similar landscape. Scaling up our measured deposition rate to all of South Central Appalachia, an area of over 94,000 km 2 and home to five million people, suggests a yearly MP deposition of approximately 321 metric tonnes. Our study highlights the prevalence of MP deposition in the Eastern United States, providing baseline data for future work to further assess routes of MP introduction., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Background: Inguinal hernias are encountered commonly, but there is a lack of uniformity and standardization in repair techniques. There are a variety of repair methods, from open methods to laparoscopic approaches. The available laparoscopic techniques that exist for inguinal hernia repair are transabdominal preperitoneal (TAPP) repair and totally extraperitoneal (TEP) repair. This study aims to compare the outcomes of these two laparoscopic procedures., Methods: The study was initiated after receiving ethical clearance. This single-centric retrospective study included the patients that underwent laparoscopic inguinal hernia repair from January 1, 2020, to December 31, 2023. Patient records were reviewed and assessed against the inclusion criteria of the study. Systematic random sampling was applied to select patients from the records. A structured questionnaire was prepared for the purpose of data collection. The data were refined and analyzed using the SPSS software., Results: A total of 200 patients (100 in each group) who underwent laparoscopic repair of an inguinal hernia were part of this study. The median (IQR) age of patients in the TAPP group was 50 (41.25-64.75) years, and that in the TEP group was 48.5 (39.25-64.75) years. The TAPP procedure took a median (IQR) time of 81 (77-88) min, while the TEP procedure took 69 (66-73) min to finish. The patients in the TAPP group returned to their normal activities after 15 (14-16) days of surgery, and the patients in the TEP group returned after 13 (11-14) days. The pain score at 1 week was found to be better in the TEP group. No difference was observed in complications or recurrence., Conclusion: TEP repair was found to be superior to TAPP repair in terms of procedural time, return to normal activities, and pain score at 1 week., Competing Interests: The authors declare no conflicts of interest., (© 2024 The Author(s). Health Science Reports published by Wiley Periodicals LLC.)

The likelihood of antimicrobial failure in COVID-19 patients with bacterial superinfection arises from both phenotypic (biofilms) and genotypic mechanisms. This cross-sectional study aimed to determine the inhibitory concentrations of quinolones-nalidixic acid, norfloxacin, ciprofloxacin, ofloxacin, and levofloxacin-in biofilm formers (minimum biofilm inhibitory concentration [MBIC]) and nonformers (minimum inhibitory concentration [MIC]) and correlate inhibitory concentrations with plasmid-mediated quinolone resistance (PMQR) genes in quinolone-resistant bacteria isolated from COVID-19 inpatients. Quinolone-resistant bacteria (n = 193), verified through disc diffusion, were tested for quinolone inhibitory concentrations using broth microdilution and biofilm formation using microtiter plate methods. The polymerase chain reaction was used to detect PMQR genes. Study variables were analyzed using SPSS v.17.0, with a significance level set at P <0.05. MIC-to-MBIC median fold increases for ciprofloxacin, ofloxacin, and levofloxacin were 128 (2-8,192), 64 (4-1,024), and 32 (4-512) in gram-positive cocci (GPC, n = 43), respectively, whereas they were 32 (4-8,192), 32 (4-2,048), and 16 (2-1,024) in fermentative gram-negative bacilli (F-GNB, n = 126) and 16 (4-4,096), 64 (2-64), and 16 (8-512) in nonfermentative gram-negative bacilli (NF-GNB, n = 24). In biofilm-forming F-GNB and NF-GNB, qnrB (10/32 versus 3/10), aac(6')-Ib-cr (10/32 versus 4/10), and qnrS (9/32 versus 0/10) genes were detected. A 32-fold median increase in the MIC-to-MBIC of ciprofloxacin was significantly (P <0.05) associated with qnrA in F-GNB and qnrS in NF-GNB. Biofilms formed by F-GNB and NF-GNB were significantly associated with the aac(6')-Ib-cr and qnrS genes, respectively. Nearly one-third of the superinfecting bacteria in COVID-19 patients formed biofilms and had at least one PMQR gene, thus increasing the need for quinolones at higher inhibitory concentrations.

Bovine tuberculosis (bTB) is a chronic zoonotic disease affecting cattle of all age groups including wild animals. It poses a significant threat to public health and high economic losses to dairy farmers. While the disease has been eradicated from most of the developed countries through extensive surveillance, testing and culling strategy, it is endemic in Africa, Asia, and the Middle East countries. Currently, there is limited research regarding the prevalence of bTB in cattle in Bhutan. This study aimed to determine the seroprevalence of bTB in cattle in six districts of eastern Bhutan. A two-stage probability proportional to size (PPS) sampling strategy was used to determine the number of animals from which serum samples needed to be collected in each district and sub-district. All farms and cattle for sampling were randomly selected from the data in the annual livestock census of 2020. The samples were tested using bTB ELISA test kit. The seroprevalence and their 95% confidence intervals were calculated. Logistic regression models were constructed to assess the influence of various individual animal and environmental risk factors (breed, age, sex, source of animal, body condition scores of animals, respiratory system status) associated with sero-positivity in animals. The study revealed an apparent seroprevalence of 2.57% (25/971 cattle; 95% CI:1.58-3.57), with an estimated true seroprevalence of 0.91% (95% CI: 0.0-2.81). However, none of the variables were found to be significantly associated with bTB seroprevalence in cattle. We recommend, further sampling and employment of confirmatory testing to fully ascertain the extent of bTB in the cattle herds in eastern Bhutan for prevention and control., Competing Interests: ‘The authors have declared that no competing interests exist., (Copyright: © 2024 Wangmo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Pecan ( Carya illinoensis ), an economically important deciduous tree, bears commercially valuable nutritional nuts. Spring freezes in April can severely injure pecan buds, decreasing bloom, and fruit set. This study determined how low temperatures affect pecan buds/flowers at different growth stages in several pecan scion/rootstock combinations. This study focused on three pecan scion/rootstock combinations: Pawnee/Peruque (PP), Kanza/Giles (KG), and Maramec/Colby (MC), grown at the Cimarron Valley Research Station, Perkins, Oklahoma. Branches at three different growth stages, i.e., outer bud scale shed, one week after bud break, and early bloom stages were collected from PP, MC, and KG. Branches were held in a Conviron E8 freezing unit at 4 temperatures (-2, 0, 2, and 4°C) for 4 and 8 hours; A total of 8 treatments. One sample set was kept as an untreated control. After 2-3 weeks, branch samples from all the temperature treatments were observed and categorized into two groups. Group one with number of branches had healthy buds/formation of healthy leaves/flowers and group two with number of dead branches. The carbohydrate content reserved from dormant was analyzed using an Anthrone reagent. Visual observations and carbohydrate analyses revealed differences in damage and carbohydrate content among the scion/rootstock combinations, low-temperature treatments, and growth stages. The MC combination had minimum visual damage to leaves, buds, and flowers and significantly lower soluble sugars and starch in bark phloem as well as significantly lower soluble sugars in woody tissue xylem. The KG combination had maximum visual damage and significantly higher soluble sugars and starches in the bark, and soluble sugars in the woody tissues. These results indicate the MC combination is more tolerant to spring freeze damage at all three growth stages compared to the other two pecan scion/rootstock combinations. The results also demonstrate the MC combination is using more non-structural carbohydrates, soluble sugars and starches, suggesting this is a possible mechanism in its freeze tolerance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Kaur, Zhang, Maness, Ferguson, Graham, Sun, Panta, Pokhrel, Yang and Moss.)

The mechanisms of hepatic stellate cell (HSC) activation and the development of liver fibrosis are not fully understood. Here, we show that deletion of a nuclear seven transmembrane protein, TM7SF3, accelerates HSC activation in liver organoids, primary human HSCs, and in vivo in metabolic-dysfunction-associated steatohepatitis (MASH) mice, leading to activation of the fibrogenic program and HSC proliferation. Thus, TM7SF3 knockdown promotes alternative splicing of the Hippo pathway transcription factor, TEAD1, by inhibiting the splicing factor heterogeneous nuclear ribonucleoprotein U (hnRNPU). This results in the exclusion of the inhibitory exon 5, generating a more active form of TEAD1 and triggering HSC activation. Furthermore, inhibiting TEAD1 alternative splicing with a specific antisense oligomer (ASO) deactivates HSCs in vitro and reduces MASH diet-induced liver fibrosis. In conclusion, by inhibiting TEAD1 alternative splicing, TM7SF3 plays a pivotal role in mitigating HSC activation and the progression of MASH-related fibrosis., Competing Interests: Declaration of interests R.I. and J.M.O. are co-inventors on a provisional patent for the use of ASO 56 as an inhibitor of liver fibrosis., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Hypereosinophilic syndrome with respiratory distress and multiorgan involvement is not so common in children. It is essential to identify this entity based on clinical, laboratory, and imaging features. Corticosteroids should be instituted at the earliest to stabilize the patient and prevent organ damage. Tropical infections are a common secondary cause in children warranting the administration of Diethylcarbamazine. We present a case of an adolescent male in respiratory distress with marked eosinophilia and organs involving the lungs (pulmonary infiltrates with effusion), heart (pericardial effusion), and abdomen (ascites with infiltrates in the liver) which was managed with steroids and anthelmintics. The case highlights the importance of identifying patients with Hypereosinophilic syndrome in pursuing thorough evaluation and commencing therapy.

Background: Low back pain due to disc herniation is a common problem causing frequent hospital visits and loss of working days with major socio-economic impact. Conservative treatments like analgesics, physiotherapy do not work in all patients. Surgical treatment has been the mainstay of treatment when indicated but is associated with anesthetic and surgical complications. Intradiscal oxygen-ozone chemonucleolysis is a minimally invasive procedure done under local anesthesia and has promising role in shrinking the bulged disc and reducing nerve root compression and related symptoms. This retrospective study was done to see how intradiscal oxygen-ozone chemonucleolysis reduces pain severity in patients with discogenic low back pain., Methods: Retrospective data were retrieved of those patients who underwent fluoroscopy guided intradiscal oxygen-ozone chemonucleolysis with 5-6 ml of an O2-O3 mixture (concentration of 30 microgram/ml) during a period of two years in Nepal pain care and research center. Numerical pain scale (NRS) at various follow ups were compared to preprocedural NRS., Results: Preprocedural NRS was 8± 13. NRS at three hours, one week, one month, three months and six months were 2± 13 (73 percent reduction), 2± 53 (68 percent reduction), 2± 27 (72 percent reduction), 1± 08 (77 percent reduction) and 1± 67 (79 percent reduction) respectively., Conclusions: Intradiscal oxygen-ozone chemonucleolysis can be a useful modality of treatment for discogenic low back pain in patients who fail to respond to conservative management and in whom surgery is not indicated.

Background: Traumatic cervical spinal injuries can severely affect respiratory function and cause significant morbidity and mortality. The typical respiratory morbidity in cervical spine injury is Atelectasis, Ventilator-associated pneumonia, acute respiratory distress syndrome and delayed weaning, etc. The study aims to see the prevalence of respiratory morbidity as well as mortality associated with cervical spine injury., Methods: Cross sectional study based on retrospective data was conducted on the X Sciences with the existing hospital record during the period of 3 years to find out the prevalence respiratory morbidity like Ventilatory Associated Pneumonia, delayed weaning, ARDS, atelectasis of traumatic cervical spine injury, determine the prevalence, type, and impact of respiratory morbidity and mortality in this population., Results: Total no 76 patients data meeting the inclusion criteria included in study. Male patients were more prone to develop traumatic cervical spinal injuries (SCI). The prevalence of respiratory morbidity in term of VAP(57.89), delayed weaning(46.05) and Atelectasis(22.36) was high. Patients with Asia A Neurology has higher association for VAP and delayed weaning, while Asia E Neurology patients had no respiratory morbidity. The study found a significant positive association between respiratory morbidity with hospital stay, and ventilator days (p-value: 0.019 and 0.048). A total of 15 patients died, 28.95% were discharged on request and 40.8% leaving the hospital against medical advice., Conclusions: The prevalence of respiratory morbidity higher in cervical spine injury. Furthermore, it has associated with prolonged ICU and ventilator days and increase in mortality.

DNA metabolic processes including replication, repair, recombination, and telomere maintenance occur on single-stranded DNA (ssDNA). In each of these complex processes, dozens of proteins function together on the ssDNA template. However, when double-stranded DNA is unwound, the transiently open ssDNA is protected and coated by the high affinity heterotrimeric ssDNA binding Replication Protein A (RPA). Almost all downstream DNA processes must first remodel/remove RPA or function alongside to access the ssDNA occluded under RPA. Formation of RPA-ssDNA complexes trigger the DNA damage checkpoint response and is a key step in activating most DNA repair and recombination pathways. Thus, in addition to protecting the exposed ssDNA, RPA functions as a gatekeeper to define functional specificity in DNA maintenance and genomic integrity. RPA achieves functional dexterity through a multi-domain architecture utilizing several DNA binding and protein-interaction domains connected by flexible linkers. This flexible and modular architecture enables RPA to adopt a myriad of configurations tailored for specific DNA metabolic roles. To experimentally capture the dynamics of the domains of RPA upon binding to ssDNA and interacting proteins we here describe the generation of active site-specific fluorescent versions of human RPA (RPA) using 4-azido-L-phenylalanine (4AZP) incorporation and click chemistry. This approach can also be applied to site-specific modifications of other multi-domain proteins. Fluorescence-enhancement through non-canonical amino acids (FEncAA) and Förster Resonance Energy Transfer (FRET) assays for measuring dynamics of RPA on DNA are also described. The fluorescent human RPA described here will enable high-resolution structure-function analysis of RPA-ssDNA interactions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Introduction: Ileocecal knot syndrome, a rare cause of small bowel obstruction where the ileum wraps around the cecum, poses a significant challenge for preoperative diagnosis. Prompt intervention is crucial due to the risk of rapid bowel deterioration and increased mortality., Case Presentation: A 45-year-old female presented with central abdominal pain associated with vomiting, abdominal distension, and obstipation. On examination, she was ill-looking with hypotension, tachycardia with a feeble pulse, direct and rebound abdominal tenderness, and absent bowel sounds. Aggressive fluid resuscitation was done. Based on the clinical presentation and abdominal radiograph suggestive of intestinal obstruction, an emergency exploratory laparotomy was done, which showed an ileocecal knot and 130 cm of gangrenous ileum. Peritoneal lavage followed by resection of non-viable ileum with double barrel ileostomy was done., Discussion: Ileosigmoid, appendico-ileal, ileoileal, and ileocecal knotting are the various types of intestinal knotting, with very few cases of ileocecal knotting being reported. Intestinal knotting causes severe bowel obstruction, resulting in reduced mucosal perfusion, progressive ischemia, and peritonitis, leading to high mortality. X-ray findings of multiple air-fluid levels are non-specific, and for definitive diagnosis, laparotomy is required. Assessing bowel viability before definitive surgery is essential. Despite positive outcomes, extensive resection can lead to malabsorption and ileus, with potential risk for developing short bowel syndrome., Conclusion: Despite its rarity, the possibility of ileocecal knotting should be considered in cases of small bowel obstruction due to its potential for rapid deterioration. Prompt resuscitation followed by emergency laparotomy is necessary to prevent mortality., Competing Interests: There are no conflicts to be declared.Sponsorships or competing interests that may be relevant to content are disclosed at the end of this article., (Copyright © 2024 The Author(s). Published by Wolters Kluwer Health, Inc.)

Clinicians face a global challenge treating infections caused by Enterobacteriaceae because of the high rate of antibiotic resistance. This cross-sectional study from the Nepal Armed Police Force Hospital, Kathmandu, Nepal, characterized resistance patterns in Enterobacteriaceae across different antimicrobial classes and assessed incidences of multidrug-resistant (MDR) and extensively drug-resistant (XDR) infections. Enterobacteriaceae from clinical samples were isolated on blood and MacConkey agar, except for urine samples on cysteine lactose electrolyte-deficient agar. To determine antimicrobial susceptibility patterns, including MDR and XDR, the Kirby-Bauer disc diffusion method was used. Statistics were performed using SPSS, v. 17.0. Members of the family were identified in 14.5% (95% CI: 16.2-12.8%) of the total samples (N = 1,617), primarily in urine (54.7%, 128/234), blood (19.7%, 46/234), and sputum (15.0%, 35/234). Escherichia coli (n = 118, 44.2%) was the most predominant bacteria, followed by Citrobacter freundii (n = 81, 30.3%). As much as 95.6% (392/410) of the isolates were penicillin-resistant, whereas only 36.2% (290/801) were carbapenem-resistant. A total of 96 (36.0%) MDR and 98 (36.7%) XDR Enterobacteriaceae were identified. Proteus mirabilis (44.4%, 8/18) predominated MDR cultures, whereas C. freundii (53.1%, 43/81) predominated XDR cultures. Multidrug resistant (38.4%, 71/154) and XDR Enterobacteriaceae (22.7%, 35/154) were chiefly uropathogens. Fluoroquinolone resistance rates in non-MDR, MDR, and XDR isolates were 19.9%, 63.2%, and 96.2%, respectively, whereas cephalosporin resistance rates were 28.6%, 72.9%, and 95.4% and penicillin resistance rates were 67.0%, 97.4%, and 98.0%. One-seventh of patients visiting the hospital were found to be infected with Enterobacteriaceae, and of these patients, at least one-fourth were infected with MDR strains.

Silvicultural systems are essential for effective forest management and utilize the resources by conserving biodiversity, health, and valuable services forests offer to society. However, Nepal faces a significant knowledge gap due to the limited data on the effect of silvicultural systems and forest management practices on tree diversity and regeneration. Therefore, this study was conducted to assess the effects of canopy opening on natural regeneration and species diversity of Sal ( Shorea robusta Gaertn. f.) forest in comparison to managed and control blocks in the southern plains of Nepal under irregular shelterwood systems. The vegetation sampling was carried out by the quadrat method (plot of 10 m* 10 m) for studying 48 plots in both managed and control forest areas. The Shannon-Wiener Index and the Simpson Index were used to measure species diversity, whereas Margalef's index was used to measure species richness. Dominant tree species in both managed and control forest areas were identified through the Important Value Index (IVI). The seedling and sapling density of Sal was found to be significantly higher (p < 0.05) in managed blocks. Seedlings of Sal from the seed origin were notably more abundant in the managed block (p < 0.01), whereas those from coppice origin exhibited higher numbers in the control block (p < 0.01). Sal was found to be dominant species in both managed block (IVI = 199.94) and control (IVI = 108.34). The species diversity and richness were significantly declining in the managed forest (p < 0.01) with higher species dominance in comparison to the control blocks. Our study found a positive correlation of canopy cover with species diversity (r = 0.69, p < 0.01) and richness (r = 0.59, p < 0.01) whereas a negative correlation with seedling (r = -0.43, p < 0.01) and sapling density (r = -0.16, p < 0.01). The application of the irregular shelterwood system in study area has effectively promoted natural Sal regeneration while concurrently reducing species diversity. To strike a balance between timber production and biodiversity conservation in these forests, further research focusing on moderating felling intensity within the irregular shelterwood system is strongly recommended., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

DNA metabolic processes including replication, repair, recombination, and telomere maintenance occur on single-stranded DNA (ssDNA). In each of these complex processes, dozens of proteins function together on the ssDNA template. However, when double-stranded DNA is unwound, the transiently open ssDNA is protected and coated by the high affinity heterotrimeric ssDNA binding Replication Protein A (RPA). Almost all downstream DNA processes must first remodel/remove RPA or function alongside to access the ssDNA occluded under RPA. Formation of RPA-ssDNA complexes trigger the DNA damage checkpoint response and is a key step in activating most DNA repair and recombination pathways. Thus, in addition to protecting the exposed ssDNA, RPA functions as a gatekeeper to define functional specificity in DNA maintenance and genomic integrity. RPA achieves functional dexterity through a multi-domain architecture utilizing several DNA binding and protein-interaction domains connected by flexible linkers. This flexible and modular architecture enables RPA to adopt a myriad of configurations tailored for specific DNA metabolic roles. To experimentally capture the dynamics of the domains of RPA upon binding to ssDNA and interacting proteins we here describe the generation of active site-specific fluorescent versions of human RPA (RPA) using 4-azido- L -phenylalanine (4AZP) incorporation and click chemistry. This approach can also be applied to site-specific modifications of other multi-domain proteins. Fluorescence-enhancement through non-canonical amino acids (FEncAA) and Förster Resonance Energy Transfer (FRET) assays for measuring dynamics of RPA on DNA are also described.

The amygdala processes positive and negative valence and contributes to addiction, but the cell-type-specific gene regulatory programs involved are unknown. We generated an atlas of single-nucleus gene expression and chromatin accessibility in the amygdala of outbred rats with high and low cocaine addiction-like behaviors following prolonged abstinence. Differentially expressed genes between the high and low groups were enriched for energy metabolism across cell types. Rats with high addiction index (AI) showed increased relapse-like behaviors and GABAergic transmission in the amygdala. Both phenotypes were reversed by pharmacological inhibition of the glyoxalase 1 enzyme, which metabolizes methylglyoxal-a GABA A receptor agonist produced by glycolysis. Differences in chromatin accessibility between high and low AI rats implicated pioneer transcription factors in the basic helix-loop-helix, FOX, SOX and activator protein 1 families. We observed opposite regulation of chromatin accessibility across many cell types. Most notably, excitatory neurons had greater accessibility in high AI rats and inhibitory neurons had greater accessibility in low AI rats., (© 2023. The Author(s).)

Homologous recombination (HR) is an essential double-stranded DNA break repair pathway. In HR, Rad52 facilitates the formation of Rad51 nucleoprotein filaments on RPA-coated ssDNA. Here, we decipher how Rad52 functions using single-particle cryo-electron microscopy and biophysical approaches. We report that Rad52 is a homodecameric ring and each subunit possesses an ordered N-terminal and disordered C-terminal half. An intrinsic structural asymmetry is observed where a few of the C-terminal halves interact with the ordered ring. We describe two conserved charged patches in the C-terminal half that harbor Rad51 and RPA interacting motifs. Interactions between these patches regulate ssDNA binding. Surprisingly, Rad51 interacts with Rad52 at two different bindings sites: one within the positive patch in the disordered C-terminus and the other in the ordered ring. We propose that these features drive Rad51 nucleation onto a single position on the DNA to promote formation of uniform pre-synaptic Rad51 filaments in HR., (© 2023. Springer Nature Limited.)

Background: Typhoid fever, an infective bacterial disease, is capable of causing fatal systemic infection in humans, and in an era of antimicrobial resistance, it has become of public health importance. This study aimed to investigate the laboratory diagnosis of Salmonella bloodstream infection, its serotype, antimicrobial resistance pattern, and seasonal variation at a tertiary care children's hospital., Methods: We undertook a retrospective, cross-sectional study by reviewing hospital-based laboratory records of patients whose blood culture samples were submitted from the outpatient department to the laboratory of a tertiary care children's hospital in Kathmandu, Nepal, from January 2017 to January 2019., Results: Among the total blood culture samples obtained ( n  = 39,771), bacterial isolates ( n  = 1,055, 2.65%) belonged either to the Genus Enterobacteriaceae or Genus Acinetobacter . Altogether ( n  = 91, 8.63%), isolates were positive for Salmonella spp., which were further identified as Salmonella enterica subsp. enterica ser. Typhi ( n  = 79, 7.49%), Salmonella enterica subsp. enterica ser. Paratyphi A ( n  = 11, 1.04%), and Salmonella enterica subsp. enterica ser. Paratyphi B ( n  = 1, 0.1%). The median age of patients was 6  years (IQR: 4-9), with male and female patients constituting ( n  = 53, 58.24%; OR, 1.0; 95% CI, 0.60-1.67) and ( n  = 38, 41.76%; OR, 0.98; 95% CI, 0.49-2.05) cases, respectively. The disease was observed throughout the year, with a high prevalence toward the spring season (March-May). An antibiogram showed resistance more toward nalidixic acid with S. Typhi, comprising half the isolates ( n  = 52, 65.82%; p  = 0.11). Resistance toward β-lactams with β-lactamase inhibitors (amoxicillin/clavulanate; 1.27%) was seen in a single isolate of S. Typhi. The multidrug resistance pattern was not pronounced. The multiple antibiotic resistance (MAR) index was in the range between 0.14 and 0.22 in S. Typhi and 0.22 and 0.23 in S. Paratyphi., Conclusion: Salmonella Typhi was the predominant ser. Infection was common among children between 1 and 5 years of age, showing male predominance and with the spring season contributing to a fairly higher number of cases. Antimicrobial susceptibility testing of S. Typhi showed more resistance toward nalidixic acid, with only a single isolate resistant to β-lactamase inhibitors (amoxicillin/clavulanate). Alarming multidrug resistance patterns were not observed. The MAR index in this study indicates the importance of the judicious use of antimicrobials and hospital infection prevention and control practices., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Pokhrel, Chapagain, Thakur, Basnet, Amatya, Singh and Ghimire.)

Objectives: Reports from other countries have indicated that severe forms and fatal cases of COVID-19 in older adults and people with underlying comorbidities. The aim of this study was to assess the risk factors associated with COVID-19 mortality in Nepal., Methods: A cross-sectional study was conducted from April 12 to July 23, 2021 to identify the underlying factors associated with COVID-19 deaths. Our sample included all cases diagnosed and registered as COVID-19-related deaths at 30 hospitals of Nepal., Results: A total of 1459 COVID-19 hospital-based death records were collected from 30 hospitals. Mean age at death was 60.2 (±15.6) years. One-third of cases were admitted with fever, cough, and shortness of breath. The computerized tomography Severity Score showed that 7.3% of the individuals who underwent high-resolution computerized tomography chest had a severe form of lung involvement, and 3.6% had mild to moderate involvement. The most common comorbidities were hypertension (43.7%) followed by diabetes mellitus (25.8%). Among the deceased, 37.7% were diagnosed as cases of COVID-19 pneumonia. The most common recorded causes of death were respiratory failure followed by cardio-pulmonary arrest., Conclusions: Individuals with comorbidities including hypertension and diabetes mellitus were at greater risk of developing complications and had a higher rate of mortality., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Published by Elsevier Ltd on behalf of International Society for Infectious Diseases.)

Homologous recombination (HR) is a pathway for the accurate repair of double-stranded DNA breaks. These breaks are resected to yield single-stranded DNA (ssDNA) that are coated by Replication Protein A (RPA). Saccharomyces cerevisiae Rad52 is a mediator protein that promotes HR by facilitating formation of Rad51 nucleoprotein filaments on RPA-coated ssDNA. Canonically, Rad52 has been described to function by displacing RPA to promote Rad51 binding. However, in vitro , Rad51 readily forms a filament by displacing RPA in the absence of Rad52. Yet, in vivo , Rad52 is essential for HR. Here, we resolve how Rad52 functions as a mediator using single-particle cryo-electron microscopy and biophysical approaches. We show that Rad52 functions as a homodecamer and catalyzes single-position nucleation of Rad51. The N-terminal half of Rad52 is a well-ordered ring, while the C-terminal half is disordered. An intrinsic asymmetry within Rad52 is observed, where one or a few of the C-terminal halves interact with the ordered N-terminal ring. Within the C-terminal half, we identify two conserved charged patches that harbor the Rad51 and RPA interacting motifs. Interactions between these two charged patches regulate a ssDNA binding. These features drive Rad51 binding to a single position on the Rad52 decameric ring. We propose a Rad52 catalyzed single-position nucleation model for the formation of pre-synaptic Rad51 filaments in HR., Competing Interests: Competing interests The authors declare no competing interests.

Errors in chromosome segregation underlie genomic instability associated with cancers. Resolution of replication and recombination intermediates and protection of vulnerable single-stranded DNA (ssDNA) intermediates during mitotic progression requires the ssDNA binding protein Replication Protein A (RPA). However, the mechanisms that regulate RPA specifically during unperturbed mitotic progression are poorly resolved. RPA is a heterotrimer composed of RPA70, RPA32 and RPA14 subunits and is predominantly regulated through hyperphosphorylation of RPA32 in response to DNA damage. Here, we have uncovered a mitosis-specific regulation of RPA by Aurora B kinase. Aurora B phosphorylates Ser-384 in the DNA binding domain B of the large RPA70 subunit and highlights a mode of regulation distinct from RPA32. Disruption of Ser-384 phosphorylation in RPA70 leads to defects in chromosome segregation with loss of viability and a feedback modulation of Aurora B activity. Phosphorylation at Ser-384 remodels the protein interaction domains of RPA. Furthermore, phosphorylation impairs RPA binding to DSS1 that likely suppresses homologous recombination during mitosis by preventing recruitment of DSS1-BRCA2 to exposed ssDNA. We showcase a critical Aurora B-RPA signaling axis in mitosis that is essential for maintaining genomic integrity., (© 2023. The Author(s).)

Background:  Brought with the advancements in transplantation science and the development of immunosuppressive agents, immunocompromised patients characterized with defective immunity have increased throughout the world with increased risk for opportunistic infections. This study provides an overview of the antimicrobial susceptibility pattern among opportunistic pathogens isolated from immunocompromised patients.  Methods: Clinical and laboratory records of immunocompromised patients [patients with chronic kidney disease neutropenia, diabetes, rheumatic heart disease acquired immune deficiency syndrome hepatitis B, hepatitis C, who were subjected to microbiological culture analysis in the Department of Clinical Microbiology, KIST Medical College and Teaching Hospital, for 2 years (January 2019 and December 2020) were analyzed., Results:  Out of 8,402 immunocompromised patients, 954 (11.4%) patients were subjected to microbiological culture analysis. Among 954 patients, 253 (26.5%) patients [median(interquartile range) age: 52(31-67) years; male 138 (54.5%)] were infected. A total of 295 pathogens were isolated from 1,331 cultured samples. Infections due to Escherichia coli (n=71, 24.1%), Klebsiella spp. (n=55, 18.6%), Acinetobacter calcoaceticus-baumannii complex (n=35, 11.9%), Candida albicans (n=30, 10.2%), and Staphylococcus aureus (n=28, 9.5%) were frequently observed. Among the bacterial isolates (n=239), 81.6% (n=195) of bacteria were β-lactamase producers, 51.0% (n=122) were multi-drug resistant, 9.2% (n=195) were extensively-drug resistant, 0.8% (n=195) were pan-drug resistant, and 35.7% (n=10) of S. aureus were methicillin-resistant Staphylococcus aureus., Conclusions:  The majority of infection in immunocompromised patients is caused by Gram-negative bacteria, and is often associated with a higher number of β-lactamase producers and multi-drug resistant organisms. Prescriptions of antibiotics on the grounds of antimicrobial stewardship might help to reduce the burden of antimicrobial resistance.

The avian embryo has a remarkable ability that allows it to suspend its development during blastulation for a long time at low temperatures, and to resume normal development when incubated. This ability is used by poultry hatcheries to store eggs prior to incubation. We have previously found that this ability correlates with the temperature during storage; embryos recover much better following prolonged storage at 12°C rather than at 18°C. However, the molecular and cellular mechanisms underlying these differences are poorly understood. To successfully resume development following storage, the embryo has to shift from the blastulation phase to gastrulation. Several genes are known to partake in the blastulation-to-gastrulation transition under normal conditions, such as the pluripotency-related genes Inhibitor of DNA Binding 2 (ID2) and NANOG that are expressed during blastulation, and the gastrulation-regulating genes NODAL and Brachyury (TBXT) . However, their expression and activity following storage is unknown. To elucidate the molecular mechanisms that initiate the ability to successfully transit from blastulation to gastrulation following storage, embryos were stored for 28 days at 12°C or 18°C, and were assessed either prior to incubation, 12, or 18 h of incubation at 37.8°C. Immediately following storage at 18°C group showed remarkable impaired morphology compared to the blastoderm of the 12°C group and of non-stored control embryos. Concurrently with these, expression of ID2 and NANOG was maintained following storage at 12°C similar to the control group, but was significantly reduced upon storage at 18°C. Nevertheless, when the 18°C-stored embryos were incubated, the morphology and the reduced genes were reverted to resemble those of the 12°C group. At variance, key gastrulation genes, NODAL and its downstream effector Brachyury ( TBXT ), which were similarly expressed in the control and the 12°C group, were not restored in the 18°C embryos following incubation. Notably, ectopic administration of Activin rescued NODAL and TBXT expression in the 18°C group, indicating that these embryos maintain the potential to initiate. Collectively, this study suggests a temperature-dependent mechanisms that direct the transition from blastulation to gastrulation. These mechanisms promote a successful developmental resumption following prolonged storage at low temperatures., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Pokhrel, Genin, Sela-Donenfeld and Cinnamon.)

Following the report of the first COVID-19 case in Nepal on 23 January 2020, three major waves were documented between 2020 and 2021. By the end of July 2022, 986 596 cases of confirmed COVID-19 and 11 967 deaths had been reported and 70.5% of the population had received at least two doses of a COVID-19 vaccine. Prior to the pandemic, a large dengue virus (DENV) epidemic affected 68 out of 77 districts, with 17 932 cases and six deaths recorded in 2019. In contrast, the country's Epidemiology and Disease Control Division reported 530 and 540 dengue cases in the pandemic period (2020 and 2021), respectively. Furthermore, Kathmandu reported just 63 dengue cases during 2020 and 2021, significantly lower than the 1463 cases reported in 2019. Serological assay showed 3.2% positivity rates for anti-dengue immunoglobulin M antibodies during the pandemic period, contrasting with 26.9-40% prior to it. Real-time polymerase chain reaction for DENV showed a 0.5% positive rate during the COVID-19 pandemic which is far lower than the 57.0% recorded in 2019. Continuing analyses of dengue incidence and further strengthening of surveillance and collaboration at the regional and international levels are required to fully understand whether the reduction in dengue incidence/transmission were caused by movement restrictions during the COVID-19 pandemic.