The ATDC5 cell line exhibits the multistep chondrogenic differentiation observed during endochondral bone formation. However, it takes up to two months to complete the process of cell expansion, insulin addition to promote differentiation and further changes in culture conditions effectively to induce hypertrophy. We sought to produce consistent chondrogenesis with significant hypertrophic differentiation with simpler conditions in a more practical time period. By adding ascorbate, the prechondrogenic proliferation phase was shortened from 21 to 7 days, with production of cartilaginous nodules during the chondrogenic phase, after insulin addition, that were greater in number and larger in size. Immunohistochemistry indicated much greater matrix elaboration and the mRNA expression of sox9, aggrecan and collagen type II were all significantly increased earlier and to a much higher degree when compared with controls. Moreover, there was a robust induction of hypertrophy: Col10a1, Runx2 and Mmp13 were all induced within 7-10 days. In conclusion, addition of ascorbate to ATDC5 cultures shortened the prechondrogenic proliferation phase, produced earlier chondrogenic differentiation, heightened gene expression and robust hypertrophic differentiation, abrogating the need for extended culture times and the changes in culture conditions. This simple modification considerably enhances the practicality of this cell line for studies of chondrogenesis.

Adenosine (A) to inosine (I) RNA editing is a hydrolytic deamination reaction catalyzed by the adenosine deaminase (ADAR) enzyme acting on double-stranded RNA. This posttranscriptional process diversifies a plethora of transcripts, including coding and noncoding RNAs. Interestingly, few studies have been carried out to determine the role of RNA editing in vascular disease. The aim of this study was to determine the potential role of ADARs in congenital heart disease. Strong downregulation of ADAR2 and increase in ADAR1 expression was observed in blood samples from congenital heart disease (CHD) patients. The decrease in expression of ADAR2 was in line with its downregulation in ventricular tissues of dilated cardiomyopathy patients. To further decipher the plausible regulatory pathway of ADAR2 with respect to heart physiology, miRNA profiling of ADAR2 was performed on tissues from ADAR2-/- mouse hearts. Downregulation of miRNAs (miR-29b, miR-405, and miR-19) associated with cardiomyopathy and cardiac fibrosis was observed. Moreover, the upregulation of miR-29b targets COL1A2 and IGF1, indicated that ADAR2 might be involved in cardiac myopathy. The ADAR2 target vascular development associated protein-coding gene filamin B (FLNB) was selected. The editing levels of FLNB were dramatically reduced in ADAR2 -/- mice; however, no observable changes in FLNB expression were noted in ADAR2 -/- mice compared to wild-type mice. This study proposes that sufficient ADAR2 enzyme activity might play a vital role in preventing cardiovascular defects.

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Objective: To determine the load, clinical and laboratory findings of the patients with decompensated cirrhosis admitted in medical department of tertiary care hospital and to make plan for the improvement of these patients. Study Design: Cross Sectional study. Setting: Department of medicine of Aziz Bhatti Shaheed Teaching Hospital, Gujrat. Period: from 1st January 2019 to 31st March 2019. Material & Methods: All 964 patients who were admitted included in study, clinical and laboratory features of patients with decompensated cirrhosis were recorded. Results: Male were in majority (53.7%), median age was 39 years. Decompensated cirrhosis was found  in  216 (22.4%), patients due to complication of  Diabetes Mellitus were 170 (17.6%), COPD & Asthma was found in 130 patients(13.5%), Stroke & Hypertension in 126 patients while Gastroenteritis 6% and infectious diseases RTI, UTI, Enteric Fever etc were present in 46 patients. Among 216 patients, one hundred & seventy eight were Hepatitis C Positive, fifteen had Hepatitis B, and eight had history of alcohol consumption. Seven patients were both Hepatitis B and C positive while in 3.6% etiology was other than mentioned above. Ascites was noted in two hundred six patients, 36.5% had variceal bleed while 51 admitted due to encephalopathy. Conclusion: HCV related cirrhosis and its complications like upper GI bleed, encephalopathy, ascites and hepatoma are major burden on our hospitals and need special attention.