Tissue Determinants of Human NK Cell Development, Function, and Residence.

Immune responses in diverse tissue sites are critical for protective immunity and homeostasis. Here, we investigate how tissue localization regulates the development and function of human natural killer (NK) cells, innate lymphocytes important for anti-viral and tumor immunity. Integrating high-dimensional analysis of NK cells from blood, lymphoid organs, and mucosal tissue sites from 60 individuals, we identify tissue-specific patterns of NK cell subset distribution, maturation, and function maintained across age and between individuals. Mature and terminally differentiated NK cells with enhanced effector function predominate in blood, bone marrow, spleen, and lungs and exhibit shared transcriptional programs across sites. By contrast, precursor and immature NK cells with reduced effector capacity populate lymph nodes and intestines and exhibit tissue-resident signatures and site-specific adaptations. Together, our results reveal anatomic control of NK cell development and maintenance as tissue-resident populations, whereas mature, terminally differentiated subsets mediate immunosurveillance through diverse peripheral sites. VIDEO ABSTRACT

Effect of Mycorrhiza on Vegetative Growth of Mandarin’s (Citrus reticulata Blanco) Seedlings

Citrus is among the top fruits which are cultivated around the world, and among citrus, Mandarin (Citrus reticulata Blanco) comes first and belongs to the family Rutaceae. A field experiment was conducted in Gorkha, Nepal, from April 2021 to July 2022, to study the effect of different doses of Mycorrhiza in the vegetative growth seedlings. The experiment was set up in a Randomized Complete Block Design (RCBD) with five treatments and four replications. The treatments were named T1: Control, T2: 3g/seedling, T3: 6g/seedling, T4: 9g/seedling, and T5: 11g/seedling, respectively. Eighteen months old seedlings of local Mandarin were uprooted and transplanted in finely pulverized and prepared seedbeds, and Mycorrhiza was added subsequently. Data was collected for plant height, number of leaves, and stem diameter. Data analysis was done through Microsoft- Excel, RStudio, and Analysis of Variance (ANOVA). Results of the study showed that plants treated with 9g of Mycorrhiza produced the highest number of measured parameters compared to plants treated with other doses. Control treatment (0g) produces the least number of plant leaves, Height, and stem diameter, and it can be concluded that a mycorrhizal dose of 9g is the best for the vegetative growth of Mandarin

Factors affecting utilization of mental health services from Primary Health Care (PHC) facilities of western hilly district of Nepal.

AimTo explore the factors affecting mental health service utilization from Primary Health Care facilities of Arghakhanchi district, a western hilly district of Nepal.BackgroundMental health service utilization has many facilitating and hindering factors present at different socio-ecological levels. Stigma and lack of awareness in the community have been identified as the major barriers for mental health service demand and access worldwide.MethodsA cross-sectional qualitative study was conducted in Arghakhanchi district of Nepal in July-August 2019 that collected information through face-to-face In-depth and Key Informant Interviews of three categories of participants selected judgmentally. Thirty-two purposively selected participants from the three categories were interviewed using validated interview guidelines. Thematic analysis was performed using RQDA package for EZR software. Validation of translated transcripts, member checking and inter-coder percent agreement were performed to maintain rigor in the study.ResultsMental health stigma and inadequate awareness were identified as major factors that caused barriers for mental health service utilization at community level. They also influenced different factors at other socio-ecological levels to act as barriers. Awareness in community along with accessibility and availability of comprehensive mental health services were recommended by the participants for increasing service utilization from Primary Health Care facilities.ConclusionIndividual, family and community awareness could help reduce and/or eliminate mental health stigma. Accessibility of health facilities and availability of comprehensive mental health services in Primary Health Care facilities could help increase service utilization from those facilities

The differentiation of ROR-γt expressing iNKT17 cells is orchestrated by Runx1.

iNKT cells are a unique lineage of T cells that recognize glycolipid presented by CD1d. In the thymus, they differentiate into iNKT1, iNKT2 and iNKT17 effector subsets, characterized by preferential expression of Tbet, Gata3 and ROR-γt and production of IFN-γ, IL-4 and IL-17, respectively. We demonstrate that the transcriptional regulator Runx1 is essential for the generation of ROR-γt expressing iNKT17 cells. PLZF-cre Runx1 cKO mice lack iNKT17 cells in the thymus, spleen and liver. Runx1-deficient iNKT cells have altered expression of several genes important for iNKT17 differentiation, including decreased expression of IL-7Rα, BATF and c-Maf and increased expression of Bcl11b and Lef1. However, reduction of Lef1 expression or introduction of an IL-7Rα transgene is not sufficient to correct the defect in iNKT17 differentiation, demonstrating that Runx1 is a key regulator of several genes required for iNKT17 differentiation. Loss of Runx1 leads to a severe decrease in iNKT cell numbers in the thymus, spleen and liver. The decrease in cell number is due to a combined decrease in proliferation at Stage 1 during thymic development and increased apoptosis. Thus, we describe a novel role of Runx1 in iNKT cell development and differentiation, particularly in orchestrating iNKT17 differentiation

Separation of plasmacytoid dendritic cells from B-cell-biased lymphoid progenitor (BLP) and Pre-pro B cells using PDCA-1.

B-cell-biased lymphoid progenitors (BLPs) and Pre-pro B cells lie at a critical juncture between B cell specification and commitment. However, both of these populations are heterogenous, which hampers investigation into the molecular changes that occur as lymphoid progenitors commit to the B cell lineage. Here, we demonstrate that there are PDCA-1(+)Siglec H(+) plasmacytoid dendritic cells (pDCs) that co-purify with BLPs and Pre-pro B cells, which express little or no CD11c or Ly6C. Removal of PDCA-1(+) pDCs separates B cell progenitors that express high levels of a Rag1-GFP reporter from Rag1-GFP(low/neg) pDCs within the BLP and Pre-pro B populations. Analysis of Flt3-ligand knockout and IL-7Rα knockout mice revealed that there is a block in B cell development at the all-lymphoid progenitor (ALP) stage, as the majority of cells within the BLP or Pre-pro B gates were PDCA-1(+) pDCs. Thus, removal of PDCA-1(+) pDCs is critical for analysis of BLP and Pre-pro B cell populations. Analysis of B cell potential within the B220(+)CD19(-) fraction demonstrated that AA4.1(+)Ly6D(+)PDCA-1(-) Pre-pro B cells gave rise to CD19(+) B cells at high frequency, while PDCA-1(+) pDCs in this fraction did not. Interestingly, the presence of PDCA-1(+) pDCs within CLPs may help to explain the conflicting results regarding the origin of these cells

Inhibition and Reduction of Biofilm Production along with Their Antibiogram Pattern among Gram-Negative Clinical Isolates

Background. Bacterial biofilm is a significant virulence factor threatening patients, leading to chronic infections and economic burdens. Therefore, it is crucial to identify biofilm production, its inhibition, and reduction. In this study, we investigated biofilm production among Gram-negative isolates and assessed the inhibitory and reduction potential of ethylene diamine tetra acetic acid (EDTA) and dimethyl sulfoxide (DMSO) towards them. In addition, we studied the antimicrobial resistance pattern of the Gram-negative isolates. Methods. Bacterial isolation and identification was done using standard microbiological techniques, following the Clinical and Laboratory Standards Institute (CLSI) guideline, 28th edition. The Kirby–Bauer disk diffusion method was used to determine the antibiotic susceptibility pattern of the isolates, and β-lactamase production was tested via the combination disk method. Biofilm formation was detected through the tissue culture plate (TCP) method. Different concentrations of EDTA and DMSO were used to determine their inhibitory and reduction properties against the biofilm. Both inhibition and reduction by the various concentrations of EDTA and DMSO were analyzed using paired t-tests. Results. Among the 110 clinical isolates, 61.8% (68) were found to be multidrug resistant (MDR). 30% (33/110) of the isolates were extended-spectrum β-lactamase (ESBL) producers, 14.5% (16/110) were metallo-β-lactamase (MBL), and 8% (9/110) were Klebsiella pneumoniae carbapenemase (KPC) producers. Biofilm formation was detected in 35.4% of the isolates. Biofilm-producing organisms showed the highest resistance to antibiotics such as cephalosporins, chloramphenicol, gentamicin, and carbapenem. The inhibition and reduction of biofilm were significantly lower (p < 0.05) for 1 mM of EDTA and 2% of DMSO. Conclusion. Isolates forming biofilm had a higher resistance rate and β-lactamase production compared to biofilm nonproducers. EDTA and DMSO were found to be potential antibiofilm agents. Hence, EDTA and DMSO might be an effective antibiofilm agent to control biofilm-associated infections

Prevalence of cardiovascular disease risk factors: A community-based cross-sectional study in a peri-urban community of Kathmandu, Nepal

Background: As a low-income country, Nepal is experiencing cardiovascular diseases as an emerging health problem. However, studies are lacking on the risk factors of cardiovascular diseases in peri-urban communities; where the socio-demographical transition is in progress. Therefore, this study aimed to identify the prevalence and socio-demographic distribution of cardiovascular disease risk factors in one of the peri-urban communities in Kathmandu, Nepal. Methods: We conducted a cross-sectional study in Sitapaila Village Development Committee, Kathmandu from February 2014 to February 2015. Altogether, 347 adults from 18 to 70 years of age were selected randomly. Data were collected through modified WHO STEPS questionnaire for non-communicable disease (NCD) risk factors survey and analyzed in SPSS V.16.0 software. Results: Mean age of the participant was 42.5 ± 13.2 years. Majority of them were female (n = 206; 59.4%), one-third (34%) represented Brahman and Chetri, and over a quarter (29.1%) did not attend school. Cardiovascular disease risk factors included smoking (17.6%), alcohol consumption (29.4%), insufficient fruit and vegetables intake (98%), insufficient physical activity (21.0%), obesity (15.3%), hypertension (34.4%), diabetes (10.5%), and high triglyceride levels (10.8%). They were significantly associated with different socio-demographic characteristics: smoking with gender, age groups and education level; alcohol consumption was with gender, age groups, ethnicity and occupation; insufficient physical activity with gender, age groups and occupation; hypertension with gender, age groups, ethnicity, education level and occupation. Conclusion: A high prevalence of cardiovascular disease risk factors and their disproportional distribution among the study population indicated an inevitable risk of cardiovascular events in near future. Keywords: CVD risk factors, Diabetes, Hypertension, NCDs, Nepal, Peri-urban communit