T-cell responses in COVID-19 survivors 6-8 months after infection: A longitudinal cohort study in Pune.

BACKGROUND: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immune response is crucial for disease management, although diminishing immunity raises the possibility of reinfection. METHODS: We examined the immunological response to SARS-CoV-2 in a cohort of convalescent COVID-19 patients in matched samples collected at 1 and 6-8 months after infection. The peripheral blood mononuclear cells were isolated from enrolled study participants and flow cytometry analysis was done to assess the lymphocyte subsets of naive, effector, central memory, and effector memory CD4+ or CD8+ T cells in COVID-19 patients at 1 and 6-8 months after infection. Immunophenotypic characterization of immune cell subsets was performed on individuals who were followed longitudinally for 1 month (n = 44) and 6-8 months (n = 25) after recovery from COVID infection. RESULTS: We observed that CD4 +T cells in hospitalized SARS-CoV-2 patients tended to decrease, whereas CD8+ T cells steadily recovered after 1 month, while there was a sustained increase in the population of effector T cells and effector memory T cells. Furthermore, COVID-19 patients showed persistently low B cells and a small increase in the NK cell population. CONCLUSION: Our findings show that T cell responses were maintained at 6-8 months after infection. This opens new pathways for further research into the long-term effects in COVID-19 immunopathogenesis.

Establishment of integration free iPSC clones, NCCSi011-A and NCCSi011-B from a liver cirrhosis patient of Indian origin with hepatic encephalopathy.

Liver cirrhosis accompanied with hepatic encephalopathy commonly causes cognitive impairment in patients. To model this disease, two independent patient specific induced pluripotent stem cell-line (iPSC) clones, NCCSi011-A and NCCSi011-B were generated by reprogramming the CD4+ T cells of an Indian male patient suffering from this chronic condition. Both clones expressed the stemness markers, formed embryoid bodies (EBs) with potential for spontaneous differentiation in to all the three lineages, exhibited normal karyotype (46, XY) and demonstrated alkaline phosphatase activity. These generated iPSC lines have potential for use in understanding biology of the disease and evaluation of drugs.

Development and characterization of two independent integration free iPSC clones NCCSi010-A and NCCSi010-B from a patient with alcoholic liver cirrhosis of Indian ethnicity.

We generated two human induced pluripotent stem cell-line (iPSC) clones, NCCSi010-A and NCCSi010-B, from a 32-year-old alcoholic cirrhosis patient with minimal hepatic encephalopathy of Indian origin by reprogramming his CD4+ T cells with integration free Sendai viral vector system. The generated iPSC clones showed high alkaline phosphatase activity, expressed pluripotency markers, possessed potential for multi-lineage differentiation and exhibited a normal karyotype (46, XY). These two-patient specific iPSC clones of alcoholic liver cirrhosis can potentially serve as models for disease modeling, drug development and organoid generation (Shah and Bataller, 2016).

Generation and characterization of three clones (NCCSi007A, NCCSi007B and NCCSi007C) of an integration free induced Pluripotent Stem Cell line from a patient with alcoholic liver cirrhosis of Indian ethnicity.

Three induced pluripotent stem cells (iPSC) clones NCCSi007-A, NCCSi007-B and NCCSi007-C were generated from CD4+T cells of a 38 years old male patient suffering from liver cirrhosis- alcoholic and minimal hepatic encephalopathy of Indian origin. The CD4+T cells of the patient were reprogrammed using integration free, Sendai viral vector system. Each of the three iPSC clones showed high alkaline phosphatase (ALP) activity, expressed pluripotency markers OCT4, SOX2, NANOG, KLF4, SSEA-4, TRA-1-60, showed normal male karyotype (46, XY) and exhibited multi-lineage differentiation.

Use of Smartphone-Based Video Directly Observed Therapy (vDOT) in Tuberculosis Care: Single-Arm, Prospective Feasibility Study.

BACKGROUND: India accounts for nearly one-quarter of the global tuberculosis (TB) burden. Directly observed treatment (DOT) through in-person observation is recommended in India, although implementation has been heterogeneous due largely to resource limitations. Video DOT (vDOT) is a novel, smartphone-based approach that allows for remote treatment monitoring through patient-recorded videos. Prior studies in high-income, low disease burden settings, such as the United States, have shown vDOT to be feasible, although little is known about the role it may play in resource-limited, high-burden settings. OBJECTIVE: The goal of the research was to assess the feasibility and acceptability of vDOT for adherence monitoring within a resource-limited, high TB burden setting of India. METHODS: We conducted a prospective, single-arm, pilot implementation of vDOT in Pune, India. Outcome measures included adherence (proportion of prescribed doses observed by video) and verifiable fraction (proportion of prescribed doses observed by video or verbally confirmed with the patient following an incomplete/unverifiable video submission). vDOT acceptability among patients was assessed using a posttreatment survey. RESULTS: A total of 25 patients enrolled. The median number of weeks on vDOT was 13 (interquartile range [IQR] 11-16). Median adherence was 74% (IQR 62%-84%), and median verifiable fraction was 86% (IQR 74%-98%). More than 90% of patients reported recording and uploading videos without difficulty. CONCLUSIONS: We have demonstrated that vDOT may be a feasible and acceptable approach to TB treatment monitoring in India. Our work expands the evidence base around vDOT by being one of the first efforts to evaluate vDOT within a resource-limited, high TB burden setting. To our knowledge, this is the first reported use of vDOT in India.

Glucose Monitoring in Critically Ill: Is Absence of "Stress Hyperglycemia" a Cause for Concern?

AIM: To study variations in glucose levels over 48 hours in critically ill patients by capillary blood glucose done on glucometer and compare the same in different categories of patients based on various diseases, as well as their correlation with sepsis and diabetes mellitus. To compare the same results in a subset of patients with the readings of continuous glucose monitoring. MATERIAL AND METHODS: We studied 50 critically-ill patients (Age≥18 years), admitted in medical ICU (on mechanical ventilation/ionotropic supports/in sepsis) in a teaching hospital in semi-urban Maharashtra. Critical illness was defined as any physiological instability leading to disability or death within minutes or hours, based on neurological assessment, respiratory system involvement and cardiovascular involvement. Capillary blood sugar levels were done 4 hourly using 'NIPRO' glucometer. Site was rotated. 5 patients had simultaneous continuous glucose monitoring, using I-Pro bio-sensor. RESULTS: Total 50 patients were included in the study. The data was collected and tabulated. Analysis showed that all critically ill patients showed some higher than normal recordings of blood sugar, which till now has been attributed to 'stress-hyperglycaemia'. This may be absent or blunted in sepsis. In the criticallyill patients with primary involvement of gastrointestinal tract, meal-unrelated fluctuations were seen. In critically-ill patients with CNS and CVS involvement, lowest BSL recordings were seen (meal unrelated) at 2 am. CONCLUSIONS: We concluded that that patients who develop hypoglycaemia may have an equally bad prognosis or even worse than those who develop hyperglycaemia during the period of critical illness. CGM devices record tissue glucose levels continuously, and may be useful as a 'tissue hypoglycaemia' alert.

An acute unusual presentation of hepatocellular carcinoma.

Hepatocellular Carcinoma (HCC) is a growing cause of mortality world over. The common risk factors include cirrhosis, viral infections, aflatoxin amongst others. Alpha Fetoprotein (AFP) levels and Ultrasonography (USG) are the preferred surveillance tools in early diagnosis of HCC. Here we present an unusual case of a young female with no known risk factors, no cirrhosis, no viral markers, and normal AFP levels who had a Acute hepatic failure eventually diagnosed as Primary Hepatocellular carcinoma.

Helicobacter pylori cagA, vacA and iceA genotypes in western Indian population of Maharashtra with varied gastroduodenal diseases.

Scarce reports relying on rapid urease test, serology and histopathology are currently known for H. pylori from Western India, Maharashtra. We investigated H. pylori genotypes at molecular level in gastro-duodenal disease population during the years 2002-2005. H. pylori presence was scored by polymerase chain reaction in the infected biopsies (n = 95) in various gastric diseases. H. pylori specific 16S rDNA gene amplification based preliminary identification coupled with protein coding gene amplification scores were assessed for the incidence. H. pylori 16S rDNA and 7 housekeeping genes were detected in all biopsies, whereas 71.18% and 28% found to be cagA positive and negative respectively. The vacA toxigenic alleles (vacA s1) and middle region subunit vac m1a were found in 54%, and 59% patients. However, the iceA1 was present in 40.06%; the iceA2 was less i.e. in 13.5% patients. The most common allelic combinations in different age groups irrespective of disease types were 13-30, 31-45, 46-60 and 61-73 were cagA-vac m1a-vacA s1-iceA1. In our analysis, PCR was found to be 100% accurate in detecting H. pylori in gastric biopsies. Among West Indian population H. pylori was found to be present, irrespective of any correlation with the genotype and gender of patients with the clinical outcome. However, the genotype incidences were related to age of the patients, wherein the age group ranging from 46 to 60 years was found be susceptible for H. pylori infection.