BackgroundThe decreased number and senescence of circulating endothelial progenitor cells (EPCs) are considered markers of vascular senescence associated with aging, atherosclerosis, and coronary artery disease (CAD) in elderly. In this study, we explore the role of vascular senescence in premature CAD (PCAD) in a developing country by comparing the numerical status and senescence of circulating EPCs in PCAD patients to controls.MethodsEPCs were measured by flow cytometry in 57 patients with angiographically documented CAD, and 57 controls without evidence of CAD, recruited from random patients ≤ 50 years of age at All India Institute of Medical Sciences. EPC senescence as determined by telomere length (EPC-TL) and telomerase activity (EPC-TA) was studied by real time polymerase chain reaction (q PCR) and PCR– ELISA respectively.ResultThe number of EPCs (0.18% Vs. 0.039% of total WBCs, p < 0.0001), and EPC-TL (3.83 Vs. 5.10 kb/genome, p = 0.009) were markedly lower in PCAD patients compared to controls. These differences persisted after adjustment for age, sex, BMI, smoking and medications. EPC-TA was reduced in PCAD patients, but was statistically significant only after adjustment for confounding factors (1.81 Vs. 2.20 IU/cell, unadjusted p = 0.057, adjusted p = 0.044).ConclusionsWe observed an association between increased vascular cell senescence with PCAD in a sample of young patients from India. This suggests that early accelerated vascular cell senescence may play an important mechanistic role in CAD epidemic in developing countries like India where PCAD burden is markedly higher compared to developed countries.

BackgroundSmall dense LDL is reported to be associated with increased coronary artery disease risk by various epidemiological studies. The gold standard for separation and identification of LDL subtypes in plasma is ultracentrifugation which is a lengthy procedure and difficult to perform. Various other methods like NMR, HPLC, gradient gel electrophoresis (GGE) have been reported for LDL sub fractionation all of which require specialized equipments and expertise. We report here a high throughput 3% polyacrylamide slab gel electrophoresis method (PASGE) for sub fractionation of LDL which was compared with GGE, a commonly used method for LDL sub fractionation.ResultsThe 3% PASGE method compared well with the GGE method There was a good correlation between LDL particle diameter identified by the PASGE and GGE (Pearson correlation coefficient = 0.950). A 100% concordance was found when samples were classified as per LDL phenotypes in subjects with A and B phenotype by the two methods with the concordance being 66% in subjects with intermediate (I) phenotype. The electrophoresis apparatus was optimized and designed for running twenty eight samples at a time compared to twelve to fourteen by the conventional PASGE and eight to twelve by disc electrophoresis.ConclusionThe rapid 3% polyacrylamide slab gel electrphoresis method developed is simple to perform, cost-effective and can be used for the identification LDL sub fractionation and phenotyping in large epidemiological studies.

Background Recombinant therapeutic anti-CD20 monoclonal antibody (mAb) is used for the treatment of non-Hodgkin’s lymphoma, a common B cell lymphoma constituting 80% of all lymphomas. Anti-CD20 mAb contains an Fc-linked biantennary glycan. Although, anti-CD20 monoclonal antibodies are being increasingly used for immunotherapy, their efficacy is limited in a section of patients with drug resistance to immunotherapy. There is a need to improve the efficacy by increasing the effector functions, such as the antibody-dependent cellular cytotoxicity (ADCC) activity of anti-CD20 monoclonal antibodies. Results We developed a simple and cost-effective approach to enhance ADCC effector activity in an in-house developed clone of anti-CD20 monoclonal antibody by increasing afucosylation in a new clone of Chinese Hamster Ovary (CHO) cells using 8X uridine, manganese, and galactose (UMG) to modulate the osmolality of the medium. The purified anti-CD20 monoclonal antibody from 8X UMG-containing medium showed a 2-fold increase in afucose content and 203% ADCC activity in comparison to control antibody. Conclusions Our study reports enhanced ADCC activity by modulating afucosylation using osmolality by altering simple feed additives in the culture medium.

Objectives Several studies has reported a reduced circulatory level and impaired functionality of EPCs in CAD patients and the role of eNOS in relation with reduced circulatory level of EPC and their impaired functionality has been revealed by in vitro and animal studies. However EPC’s eNOS gene expression profile in in vivo condition in PCAD patients is yet to be revealed as the prevalence of CAD at young age is markedly increased in developing countries. Our previous study has already reported a significantly reduced circulatory level of EPC in PCAD patients compared to control subjects and in continuation of that finding, present study aimed to investigate the eNOS gene expression of EPC in same study subjects as well as to establish the association between EPC’s eNOS gene expression and reduced circulatory level of EPC in PCAD patients. Results Reduced eNOS gene expression in EPC from PCAD patients compared to healthy controls were found (0.998±0.096/1.063±0.107) with a p-value of 0.002 and this difference was persistent even after adjusting for confounding factors (p=0.002). A positive correlation was found between eNOS gene expression and level of CD34+/KDR+ cells in circulation (r = 0.234 and p = 0.0664); data from previous study.

Abstract Objectives: Several studies has reported a reduced circulatory level and impaired functionality of EPCs in coronary artery disease (CAD) patients and the role of endothelial nitric oxide Synthase (eNOS) in relation with reduced circulatory level of EPC and their impaired functionality has been revealed by in vitro and animal studies. However EPC’s eNOS gene expression profile in in vivo condition in PCAD patients is yet to be revealed as the prevalence of CAD at young age is markedly increased in developing countries. Our previous study has already reported a significantly reduced circulatory level of EPC in PCAD patients compared to control subjects and in continuation of that finding, present study aimed to investigate the eNOS gene expression of EPC in same study subjects as well as to establish the association between EPC’s eNOS gene expression and reduced circulatory level of EPC in PCAD patients. Results: Reduced eNOS gene expression in EPC from PCAD patients compared to healthy controls were found (0.998±0.096/1.063±0.107) with a p-value of 0.002 and this difference was persistent even after adjusting for confounding factors (p=0.002). A positive correlation was found between eNOS gene expression and level of EPC in circulation (r = 0.234 and p = 0.0664); data from previous study.