Identification of genetic variants associated with a wide spectrum of phenotypes clinically diagnosed as Sanfilippo and Morquio syndromes using whole genome sequencing.

Mucopolysaccharidoses (MPSs) are inherited lysosomal storage disorders (LSDs). MPSs are caused by excessive accumulation of mucopolysaccharides due to missing or deficiency of enzymes required for the degradation of specific macromolecules. MPS I-IV, MPS VI, MPS VII, and MPS IX are sub-types of mucopolysaccharidoses. Among these, MPS III (also known as Sanfilippo) and MPS IV (Morquio) syndromes are lethal and prevalent sub-types. This study aimed to identify causal genetic variants in cases of MPS III and MPS IV and characterize genotype-phenotype relations in Pakistan. We performed clinical, biochemical and genetic analysis using Whole Genome Sequencing (WGS) in 14 Pakistani families affected with MPS III or MPS IV. Patients were classified into MPS III by history of aggressive behaviors, dementia, clear cornea and into MPS IV by short trunk, short stature, reversed ratio of upper segment to lower segment with a short upper segment. Data analysis and variant selections were made based on segregation analysis, examination of known MPS III and MPS IV genes, gene function, gene expression, the pathogenicity of variants based on ACMG guidelines and in silico analysis. In total, 58 individuals from 14 families were included in the present study. Six families were clinically diagnosed with MPS III and eight families with MPS IV. WGS revealed variants in MPS-associated genes including NAGLU, SGSH, GALNS, GNPTG as well as the genes VWA3B, BTD, and GNPTG which have not previously associated with MPS. One family had causal variants in both GALNS and BTD. Accurate and early diagnosis of MPS in children represents a helpful step for designing therapeutic strategies to protect different organs from permanent damage. In addition, pre-natal screening and identification of genetic etiology will facilitate genetic counselling of the affected families. Identification of novel causal MPS genes might help identifying new targeted therapies to treat LSDs.

Epidemiology and clinical spectrum of pediatric patients with human immunodeficiency virus (HIV) infection: A 15 years' experience.

Objective: To view the different patterns of presentation of HIV in pediatric population along with mode of transmission and associated co infections and co morbidities. Methods: It was a retrospective study conducted at Pakistan Institute of Medical Sciences, Islamabad, in which we evaluated the records of pediatric patients diagnosed with HIV from 2005 to 2020. All the data like age, gender, area, presenting complaints, examination findings at the time of diagnosis, mode of transmission, co infection and co morbidities were recorded. Descriptive analysis was done to calculate frequencies and means of the variables. SPSS 20 was used for data analysis. Results: Ninety four participants were evaluated with male to female ratio as 1.8:1 and mean age of 5.2 years. Majority of patients (44%) were below 4 years. Fever (55%) was the most reported symptom followed by cough (39%), diarrhoea (29%), pallor (27%), shortness of breath (26%), weight loss (23%) and failure to thrive (22%). Co infection with TB was present in (16%). Eight (9%) patients were thalassaemic. Mother to child transmission (60%) was the commonest mode of transmission followed by blood transfusion (23%) and parenteral transmission (6%). Conclusion: In children HIV is more prevalent in males especially under 4 years with fever, cough, diarrhea and pallor being the common symptoms at presentation. Tuberculosis is the commonest co infection as we are endemic for TB and mother to child transmission is the commonest mode of transmission as there was no outbreak in our area.

Identifying the genetic causes of phenotypically diagnosed Pakistani mucopolysaccharidoses patients by whole genome sequencing.

Background: Lysosomal storage disorders (LSDs) are a group of inherited metabolic diseases, which encompass more than 50 different subtypes of pathologies. These disorders are caused by defects in lysosomal enzymes, transporters, and other non-lysosomal proteins. Mucopolysaccharidosis (MPS) is the most common subgroup of lysosomal storage disorders in which the body is unable to properly breakdown mucopolysaccharides. The aim of the present study was to identify novel genes and pathogenic variants in families from diverse regions of Pakistan with clinically diagnosed mucopolysaccharidosis type I and mucopolysaccharidosis type II. Methods: Clinical diagnosis identified 12 with mucopolysaccharidosis I and 2 with mucopolysaccharidosis II in 14 families and whole genome sequencing (WGS) was performed to identify the causative variations in 15 affected individuals. Twenty-two unaffected individuals including parents or normal siblings of patients were also sequenced. Putative causal variants were identified by co-segregation and functional annotation. Results: Analysis of whole genome sequencing data revealed ten novel and six previously reported variants in lysosomal storage disorders-associated genes (IDUA, GALNS, SGSH, GAA, IDS, ALDOB, TRAPPC4, MASP1, SMARCAL, KIAA1109, HERC1, RRAS2) and a novel candidate gene (ABCA5) for lysosomal storage disorder-like phenotypes, which has previously been associated with symptoms strongly related with lysosomal storage disorder in animal models. Conclusion: Multigenic inheritance was found in several families highlighting the importance of searching for homozygous pathogenic variants in several genes also in families with a high degree of consanguinity.

Occurrence of Diarrheal Disease among Under-Five Children and Associated Sociodemographic and Household Environmental Factors: An Investigation Based on National Family Health Survey-4 in Rural India.

Diarrheal disease is a significant public health problem leading to mortality and morbidity among children aged 0-59 months in rural India. Therefore, the rationale of this study was to identify the sociodemographic and environmental predictors associated with diarrhea among under-five children in rural India. A total of 188,521 living children (0-59 months) were studied from the National Family Health Survey-4, (NFHS-4) 2015-2016. Bivariate and binary logistic regression models were carried out from the available NFHS-4 data for selected sociodemographic and environmental predictors to identify the relationship of occurrence of diarrhea using STATA 13.1. In rural India, children aged 12-23 months, 24-35 months, 36-47 months, and 48-59 months were significantly improbable to suffer diarrheal disease. Children of the female sex, as well as children of scheduled tribes (ST) and other backward classes (OBC), were less likely to experience diarrhea. The disease was more likely to occur among children of scheduled castes (SC); Muslim or other religions; children belonging to central, eastern, and western regions; children with low birth weight; as well as children with improper stool disposal and rudimentary roof materials. In the rural parts of India, sociodemographic and household environmental factors were most influential. Effective community education; improved handwashing practices; pure water supply; and proper waste disposal, including building and utilizing latrines, would help reduce the burden of diarrheal disease in children.

Implementation Of National Institute Of Health Guidelines And Other Factors Contributing To Work-Related Burnout In Covid Isolation Ward And ICU Physicians.

BACKGROUND: During the pandemic, disruption of work schedule and constant health risks causes physician burnout. This study aims to identify Burnout in physicians working in COVID ICU and isolation Ward. METHODS: A cross-sectional study was conducted on 200 physicians who had worked in either COVID ICU or Isolation ward. A survey of 23 questions was designed to assess the work-related Burnout using the Copenhagen burnout inventory and the implementation of NIH guidelines. RESULTS: Among the 200 physicians, 151 (75.5%) were well informed of the guidelines, of which 52 (34.4%) participants believed the guidelines were not sufficient. These 34.4% of individuals showed a mean burnout score of 70.05% (p-value 0.001). Medical registrars and Medical Officers suffered the highest burnout mean percentage score of 76% and 72.42%, respectively. 89 (44.5%) individuals arranged PPE suits on their own and suffered a burnout score of 71.3% ±14.35 (p-value <0.001). Seventy-two (36%) claimed their administration was not cooperative in resolve safety issues. These individuals showed a mean burnout score of 74.3% ±13.82 (p-value <0.001). CONCLUSIONS: Lack of physician's faith in the adequacy of the NIH guidelines has been found to be a significant factor in contributing to work-related Burnout. Due to the shortage of PPE, a majority of physicians have to reuse their PPE multiple times and even have to personally arrange their PPE suits, which further compounds the burnout. Increase administrative cooperation in resolving issues related to safety measures should help elevate work-related burnout in physicians working in the ICU and isolation ward.

Quantifying Serum Derived Differential Expressed and Low Molecular Weight Protein in Breast Cancer Patients.

BACKGROUND: Searching the biomarker from complex heterogeneous material for early detection of disease is a challenging task in the field of biomedical sciences. OBJECTIVE: The study has been arranged to explore the proteomics serum derived profiling of the differential expressed and low molecular weight protein in breast cancer patient. METHODS: Quantitative proteome was analyzed using the Nano LC/Mass and Bioinformatics tool. RESULTS: This quantification yields 239 total protein constituting 29% of differentially expressed protein, with 82% downregulated differential protein and 18% up-regulated differential protein. While 12% of total protein were found to be cancer inducing proteins. Gene Ontology (GO) described that the altered proteins with 0-60 kDa mass in nucleus, cytosol, ER, and mitochondria were abundant that chiefly controlled the RNA, DNA, ATP, Ca ion and receptor bindings. CONCLUSION: The study demonstrate that the organelle specific, low molecular weighted proteins are significantly important biomarker. That act as strong agents in the prognosis and diagnosis of breast cancer at early stage.