Case report: Deep sequencing and long-read genome sequencing refine prior genetic analyses in families with apparent gonadal mosaicism in PIK3CD-related activated PI3K delta syndrome.

Gonadal and gonosomal mosaicism describe phenomena in which a seemingly healthy individual carries a genetic variant in a subset of their gonadal tissue or gonadal and somatic tissue(s), respectively, with risk of transmitting the variant to their offspring. In families with one or more affected offspring, occurrence of the same apparently de novo variants can be an indicator of mosaicism in either parent. Panel-based deep sequencing has the capacity to detect low-level mosaic variants with coverage exceeding the typical limit of detection provided by current, readily available sequencing techniques. In this study, we report three families with more than one affected offspring with either confirmed or apparent parental gonosomal or gonadal mosaicism for PIK3CD pathogenic variants. Data from targeted deep sequencing was suggestive of low-level maternal gonosomal mosaicism in Family 1. Through this approach we did not detect pathogenic variants in PIK3CD from parental samples in Family 2 and Family 3. We conclude that mosaicism was likely confined to the maternal gonads in Family 2. Subsequent long-read genome sequencing in Family 3 showed that the paternal chromosome harbored the pathogenic variant in PIK3CD in both affected children, consistent with paternal gonadal mosaicism. Detection of parental mosaic variants enables accurate risk assessment, informs reproductive decision-making, and provides helpful context to inform clinical management in families with PIK3CD pathogenic variants.

Analysis of neuropathological comorbid conditions in elderly patients with mild cognitive impairment in a tertiary care center in South India.

Introduction: Mild cognitive impairment (MCI) is a transitional stage in the continuum of cognitive decline. Multiple risk factors may be involved apart from neuropathological states such as Alzheimer's disease, Parkinson's disease, and vascular dementia. There is scant data in the literature pertaining to our study population in Kerala, South India that provide associations between suggested risk factors and MCI. Most of the elderly present to family and primary care physicians with complaints of some form of memory impairment. Objectives: To find out the significant neuropathological comorbid conditions present in elderly patients with MCI. To assess for other risk factors in the same population- including laboratory parameters, comorbidities, and psychosocial parameters. Methods: This retrospective record-based study included a sample of 93 patients with MCI as quantified by the Mini-Mental Status Examination (MMSE). These subjects were compared with controls (n = 97) without MCI, with respect to neuropathological diagnoses, laboratory parameters and psychosocial parameters. Results: The findings of our study were that female gender, higher depression scores, a greater number of medications taken, benzodiazepine use, higher alkaline phosphatase levels, positive fall history, loss of a spouse, and lower levels of education were associated with MCI. MCI is negatively associated with positive alcohol history. The most commonly seen proven neuropathological diagnosis was Parkinson's disease. Conclusion: The risk factors that were found in our study should be highlighted in the elderly and preventive measures should be taken to prevent the downward progression through the cognitive continuum. Prospective studies looking into mild cognitive impairment with better screening tools and proper assessment of neuropathological comorbid conditions can further elucidate the findings related to this study.

Release of stem cells from quiescence reveals gliogenic domains in the adult mouse brain.

Quiescent neural stem cells (NSCs) in the adult mouse ventricular-subventricular zone (V-SVZ) undergo activation to generate neurons and some glia. Here we show that platelet-derived growth factor receptor beta (PDGFRß) is expressed by adult V-SVZ NSCs that generate olfactory bulb interneurons and glia. Selective deletion of PDGFRß in adult V-SVZ NSCs leads to their release from quiescence, uncovering gliogenic domains for different glial cell types. These domains are also recruited upon injury. We identify an intraventricular oligodendrocyte progenitor derived from NSCs inside the brain ventricles that contacts supraependymal axons. Together, our findings reveal that the adult V-SVZ contains spatial domains for gliogenesis, in addition to those for neurogenesis. These gliogenic NSC domains tend to be quiescent under homeostasis and may contribute to brain plasticity.

A positive attitude among primary healthcare providers predicts better hepatitis B prevention practices: evidence from a cross-sectional survey in Wakiso district, Central Uganda.

Background: Hepatitis B Virus (HBV) infection is an important occupational health risk among primary healthcare providers (PHCPs). However, there is limited evidence on whether PHCPs' level of knowledge and attitude can predict better HBV infection prevention practices. This study established the relationship between knowledge, attitude, and HBV infection prevention practices among PHCPs in Wakiso district, Central Uganda. Methods: A cross-sectional study design was used. Data were collected from 306 PHCPs, using a structured questionnaire. PHCPs were randomly selected from 55 healthcare facilities. STATA version 14.0 was used to analyse data. A 'modified Poisson' regression model was used for inferential statistics. Results: About 42.2% of PHCPs exhibited poor knowledge of HBV infection transmission and prevention, 41.8% had a negative attitude, and 41.5% exhibited poor prevention practices. Age (PR 1.82, 95% CI: 1.24-2.66) was positively associated with the level of knowledge. Healthcare facility level (PR 0.53, 95% CI: 0.34-0.84), main department of work (PR 0.69, 95% CI: 0.51-0.95), years in service (PR 0.66, 95% CI: 0.44-0.99), working in private not-for-profit healthcare facilities (PR 0.59, 95% CI: 0.34-0.99), and public healthcare facilities (PR 0.58, 95% CI: 0.42-0.80) were negatively associated with the level of knowledge. There was a negative association between the location of healthcare facility (PR 0.76, 95% CI: 0.62-0.93) and attitude, and a positive association between level of knowledge (PR 1.36, 95% 1.12-1.65) and attitude. Working in a public healthcare facility (PR 0.80, 95% CI: 0.64-0.99) was negatively associated with practices while having a positive attitude (PR 1.60, 95% CI: 1.28-1.99) predicted better HBV infection prevention practices. Conclusion: PHCPs who were more knowledgeable about HBV infection were more likely to have a positive attitude. In turn, having a positive attitude was associated with better HBV infection prevention practices. There is a need to sensitise PHCPs on HBV infection, and provision of screening and vaccination services in order to address the KAP gaps.

Impact of nutritional education in malnourished elderly patients with the comparison of Mini nutritional assessment (MNA) score.

INTRODUCTION: Prevalnce of malnutrition is high in the elderly. Very few studies exists mentioning the prevalnce of malmutrtion in elderly. In india around 7.7% of the total population belong to elderly age group and in kerala older adults constitute 13% of its population. The health of the elderly contributes to the health status of the population. Malnutrition is commonly seen in elders. There are multiple factors contributing to malnutrtion in this age group. MATERIAL AND METHODS: A sample of 30 patients, 60 years and above fulfilling the inclusion criteria are enrolled in the study. Malnutrition was assessed by MNA score. The score ranges from 0 to 30 points. The score above 24 show patient that are healthy with no nutritional problem, scores within range 17-23.5 indicate patients vulnerable to malnutrition and score below 17 as malnourished. MNA scoring was repeated after 3 months giving the dietary education. RESULTS: 29 patients were found to be at risk of malnutrition and 1 patient was found to be malnourished After 3 months of dietary intervention, 23 attained normal. CONCLUSION: MNA is a very accurate and effective tool for identifying malnutrition in elderly. Nutrition is an important area that need to be addressed in elder population.

Hepatitis B screening and vaccination status of healthcare providers in Wakiso district, Uganda.

BACKGROUND: Screening and vaccination against Hepatitis B virus (HBV) infection remains the most effective intervention in curbing the disease. However, there is limited evidence on the factors associated with the uptake of these services in Uganda. This study determined the uptake of HBV screening and vaccination status, and associated factors among Healthcare Providers (HCPs) in Wakiso district, Uganda. MATERIALS AND METHODS: This cross-sectional study was conducted among 306 HCPs, randomly selected from 55 healthcare facilities. Prevalence ratios (PR) were used to determine the factors associated with HBV screening and vaccination status of HCPs. RESULTS: Of the 306 HCPs, 230 (75.2%) had ever screened for HBV infection while 177 (57.8%) were fully vaccinated. Being male was positively associated with 'ever been screened' for HBV infection (Adjusted PR = 1.27, 95%CI 1.13-1.41). Working in a public healthcare facility (Adjusted PR = 0.78, 95%CI 0.68-0.90) was negatively associated with ever been screened. Male sex (Adjusted PR = 1.21, 95%CI 1.01-1.46), the belief that the HBV vaccine was safe (Adjusted PR = 1.72, 95%CI 1.03-2.89) and ever been screened (Adjusted PR = 2.28, 95%CI 1.56-3.34) were positively associated with being fully vaccinated. However, working in a public healthcare facility (Adjusted PR = 0.79, 95%CI 0.64-0.98), self-perceived risk of HBV infection (Adjusted PR = 0.72, 95% CI:0.62-0.84), and working in a healthcare facility with infection control guidelines (Adjusted PR = 0.79, 95%CI 0.66-0.95) were negatively associated with being fully vaccinated. CONCLUSION: Three quarters of HCPs had ever been screened for HBV while slightly more than half were fully vaccinated. HBV screening and vaccination interventions need to consider the HCP sex, risk perception, attitude towards safety and efficacy of the hepatitis B vaccine, and healthcare facility characteristics such as ownership and availability of infection control guidelines, in order to be successful.

Hypothalamic regulation of regionally distinct adult neural stem cells and neurogenesis.

Neural stem cells (NSCs) in specialized niches in the adult mammalian brain generate neurons throughout life. NSCs in the adult mouse ventricular-subventricular zone (V-SVZ) exhibit a regional identity and, depending on their location, generate distinct olfactory bulb interneuron subtypes. Here, we show that the hypothalamus, a brain area regulating physiological states, provides long-range regionalized input to the V-SVZ niche and can regulate specific NSC subpopulations. Hypothalamic proopiomelanocortin neurons selectively innervate the anterior ventral V-SVZ and promote the proliferation of Nkx2.1+ NSCs and the generation of deep granule neurons. Accordingly, hunger and satiety regulate adult neurogenesis by modulating the activity of this hypothalamic-V-SVZ connection. Our findings reveal that neural circuitry, via mosaic innervation of the V-SVZ, can recruit distinct NSC pools, allowing on-demand neurogenesis in response to physiology and environmental signals.

Recovery in Schizophrenia: What Consumers Know and Do Not Know.

The recovery model has permeated mental health systems by leading to the development of new psychiatric interventions and services and the reconfiguration of traditional ones. There is growing evidence that these interventions and services confer benefits in clinical and recovery-oriented outcomes. Despite the seeming adoption of recovery by policy makers, the transformation of mental health systems into recovery-oriented systems has been fraught with challenges.

Prospective identification and purification of quiescent adult neural stem cells from their in vivo niche.

Adult neurogenic niches harbor quiescent neural stem cells; however, their in vivo identity has been elusive. Here, we prospectively isolate GFAP(+)CD133(+) (quiescent neural stem cells [qNSCs]) and GFAP(+)CD133(+)EGFR(+) (activated neural stem cells [aNSCs]) from the adult ventricular-subventricular zone. aNSCs are rapidly cycling, highly neurogenic in vivo, and enriched in colony-forming cells in vitro. In contrast, qNSCs are largely dormant in vivo, generate olfactory bulb interneurons with slower kinetics, and only rarely form colonies in vitro. Moreover, qNSCs are Nestin negative, a marker widely used for neural stem cells. Upon activation, qNSCs upregulate Nestin and EGFR and become highly proliferative. Notably, qNSCs and aNSCs can interconvert in vitro. Transcriptome analysis reveals that qNSCs share features with quiescent stem cells from other organs. Finally, small-molecule screening identified the GPCR ligands, S1P and PGD2, as factors that actively maintain the quiescent state of qNSCs.

Synthesis and antimycobacterial activity of highly functionalized tetrahydro-4(1H)-pyridinones.

A series of 35 2e,3e,6e-triaryltetrahydro-4(1H)-pyridinones, 2e,3e,5e,6e-tetraaryltetrahydro-4(1H)-pyridinones and their N-nitroso and N-cyano analogs have been prepared. All these 35 compounds obtained were screened for their in vitro activity against Mycobacterium tuberculosis H37Rv (MTB). Among them, the N-nitrosopyridinones are found to be more active against MTB than the corresponding N-CN analogs, which, in turn, were slightly more active than NH analogs. In particular, the N-nitroso compounds, 3d, 4b and 4e with halogen-bearing phenyl rings at 2,6-positions showed maximum activity with MIC values of 3.97, 3.11 and 3.11 µM, being more efficacious than the first line anti-TB drugs, ciprofloxacin, ethambutol and pyrazinamide. A general trend has also been discerned in all the three classes of NH, N-CN and N-NO compounds, in each of which those bearing four aryl rings display higher activity than that having three analogously substituted aryl rings disclosing that lipophilicity could be an important factor underlying antimycobacterial activity.

Development and evaluation of a new method for sampling and monitoring the symphylid population in pineapple.

BACKGROUND: Symphylids (Hanseniella sp.) are polyphagous soilborne parasites. Today, symphylid populations on pineapple are monitored by observing root symptoms and the presence of symphylids at the bottom of basal leaves. The authors developed a reliable method with a bait and trap device to monitor symphylid populations in pineapple or fallow crops. The spatial distribution of the symphylid populations was evaluated using the variance/mean ratios and spatial analyses based on Moran's and Geary's indices. The method has been tested to monitor symphylid populations at different developmental stages of pineapple. RESULTS: Adding potato baits to the soil samples increased the trapping efficiency of symphylids when compared with 'soil only' and 'bait only' methods. The handling of the samples is also facilitated by the new device. Results showed that the vertical distribution of symphylids may be uniform deeply inside the soil profile under pineapple, up to 50 cm. Results showed that symphylid populations are highly aggregated, showing a spot area about 4-6 m wide for their development. CONCLUSION: The new method allows better and easier evaluation of symphylid populations. It may be very useful in the evaluation of new IPM methods to control symphylids under pineapple.

Endogenous biotin as a marker of adrenocortical cells with steroidogenic potential.

Interpretation of adrenal cortex phenotypes is greatly facilitated by simultaneous examination of multiple markers at single cell resolution. However, the availability of multiple appropriate antibodies can be rate limiting, while their cognate antigens are often subject to variable accessibility. Specific markers not subject to these constraints thus have obvious utility. Here we report that endogenous biotin, when detected in fixed, frozen tissue sections using fluorescent streptavidin, is a specific marker of apparently all cells with steroidogenic potential in the murine adrenal cortex. While streptavidin stains presteroidogenic and mature cortical cells, it does not label the adrenal capsule, medulla or vascular endothelium. Developmental profiles reveal adrenal endogenous biotin labeling from E13.5 through adulthood. Comparisons with zonal markers, hypothalamic-pituitary-adrenal (HPA) axis-remodeled tissue, transgenic Shh-nLacZ or Gli1-nLacZ animals, and Shh mutant embryos further demonstrate the utility of this approach. Fluorescent streptavidin applied using a simple one-step staining protocol thus provides a potent counterstain for use in adrenal analyses.

Polyphenol oxidase and peroxidase expression in four pineapple varieties (Ananas comosus L.) after a chilling injury.

Pineapple internal browning (IB) is a chilling injury that produces enzymatic browning associated with flesh translucency. Pineapple biodiversity allowed the investigation of how polyphenol oxidase (PPO) and peroxidase (POD) activities with their different isoforms are involved in the IB mechanism. Fruits of four varieties that expressed IB symptoms differently, Smooth Cayenne (SCay) and the hybrids MD2, Flhoran 41 (Flh 41), and Flhoran 53 (Flh 53), were stressed by cold. The susceptible varieties showed classical brown spots but different patterns of IB, whereas MD2 and controls showed no IB. Enzymatic activities were measured on fruit protein extracts and PPO and POD isoforms separated on mini-gels (PhastSystem). Only PPO activity was significantly enhanced in the presence of IB. Up to six PPO isoforms were identified in the susceptible varieties. PPO was barely detectable in the nonsusceptible variety MD2 and in controls. The number of PPO isoforms and the total PPO activity after chilling are varietal characteristics.

Localization of Sonic hedgehog secreting and receiving cells in the developing and adult rat adrenal cortex.

Sonic hedgehog signaling was recently demonstrated to play an important role in murine adrenal cortex development. The organization of the rat adrenal differs from that of the mouse, with the zona glomerulosa and zona fasciculata separated by an undifferentiated zone in the rat, but not in the mouse. In the present study we aimed to determine the mRNA expression patterns of Sonic hedgehog and the hedgehog signaling pathway components Patched-1 and Gli1 in the developing and adult rat adrenal. Sonic hedgehog expression was detected at the periphery of the cortex in cells lacking CYP11B1 and CYP11B2 expression, while signal-receiving cells were localized in the overlying capsule mesenchyme. Using combined in situ hybridization and immunohistochemistry we found that the cells expressing Sonic hedgehog lie between the CYP11B2 and CYP11B1 layers, and thus Sonic hedgehog expression defines one cell population of the undifferentiated zone.

A highly atom economic, chemo-, regio- and stereoselective synthesis and evaluation of spiro-pyrrolothiazoles as antitubercular agents.

The 1,3-dipolar cycloaddition of azomethine ylides derived from substituted isatins and 1,3-thiazolane-4-carboxylic acid to a series of 1-methyl-3,5-bis[(E)-arylmethylidene]-tetrahydro-4(1H)-pyridinones afforded novel spiro-pyrrolothiazoles chemo-, regio- and stereoselectively in quantitative yields. These compounds were screened for their in vitro activity against Mycobacterium tuberculosis H37Rv (MTB) and multi-drug resistant M. tuberculosis (MDR-TB) using agar dilution method. Among the synthesized compounds, spiro[5.3'']-5''-nitrooxindole-spiro-[6.3']-1'-methyl-5'-(2,4-di-chlorophenylmethylidene)tetrahydro-4'(1H)-pyridinone-7-(2,4-dichlorophenyl)tetra-hydro-1H-pyrrolo[1,2-c][1,3]thiazole (9k) was found to be the most active with a minimum inhibitory concentration (MIC) of 0.6microM against MTB and MDR-TB.

Shh signaling regulates adrenocortical development and identifies progenitors of steroidogenic lineages.

The adrenal cortex is a critical steroidogenic endocrine tissue, generated at least in part from the coelomic epithelium of the urogenital ridge. Neither the intercellular signals that regulate cortical development and maintenance nor the lineage relationships within the adrenal are well defined. We have explored adrenal Shh activity and found that Shh is expressed in relatively undifferentiated steroidogenic cells, which signal to the overlying capsule and subjacent nonsteroidogenic mesenchyme cells that we also find are progenitors of steroidogenic lineages. Shh-expressing cells also generate all steroidogenic cell types, but not nonsteroidogenic ones. Shh mutant adrenals have a thin capsule and small cortex. Our findings both support a novel dual lineage, Shh-independent and Shh-dependent, model of adrenocortical development, and identify distinct populations of adrenocortical progenitor and candidate stem cells.