Developing and testing models to predict mortality in the general population.

We have previously proposed an approach using information collected from published reports to generate prediction models. The goal of this project was to validate this technique to develop and test various prediction models. A risk indicator (R) is calculated as a linear combination of the hazard ratios for the following predictors: age, male gender, diabetes, albuminuria, and either CKD, CVD or both. We developed a linear and two exponential expressions to predict the probability of the outcome of 2-year mortality and compared to actual outcome in the target dataset from NHANES. The risk indicator demonstrated good performance with area under ROC curve of 0.84. The linear and two exponential expressions generated similar predictions in the lower categories of risk indicator (R ≤ 6). However, in the groups with higher R value, the linear expression tends to predict lower, and the exponential expressions higher, probabilities than the observed outcome. A Combined model which averaged the linear and logistic expressions was shown to approximate the actual outcome data the best. A simple technique (named Woodpecker™) allows derivation functional prediction models and risk stratification tools from reports of clinical outcome studies and their application to new populations by using only summary statistics of the new population.

Synthesis of novel amino and acetyl amino-4-methylcoumarins and evaluation of their antioxidant activity.

The six novel 4-methylcoumarins bearing different functionalities such as amino, hydroxy, N-acetyl, acetoxy and nitro have been synthesized and confirmed on the basis of their spectral data (1H-, 13C-NMR, UV, IR and EI mass). They were examined for the first time for their effect on NADPH dependent liver microsomal lipid peroxidation in vitro, and the results were compared with other model 4-methylcoumarin derivatives to establish the structure-activity relationship. Our studies demonstrated that amino group is an effective substitute for the hydroxyl group for antioxidant property and produced a dramatic inhibition of lipid peroxidation. Ortho dihydroxy and ortho hydroxy-amino coumarins were found to possess highest antioxidant and radical scavenging activities.

Cardiovascular risk assessment before and after kidney transplantation.

Cardiovascular disease (CVD) is the leading cause of death in dialysis patients and the most common cause of death and allograft loss among kidney transplant recipients. End-stage renal disease (ESRD) is associated with an increased incidence and prevalence of a wide range of CVDs including coronary artery disease, stroke, congestive heart failure, atrial fibrillation, sudden cardiac death, pulmonary hypertension, and valvular heart disease. CVD risk factors are very common in patients with ESRD, and most patients have multiple risk factors. Kidney transplantation is the treatment of choice for patients with ESRD, as a successful transplant improves longevity and quality of life, primarily by decreasing the incidence and severity of CVD. Correction of the uremic state and improved glomerular filtration rate seem to be the major mechanism of this benefit. Transplant candidates should undergo cardiovascular assessment, usually echocardiography and exercise stress testing, and may require formal cardiology consultation. Higher risk candidates, including those aged >50 years, hypertension, diabetes, established coronary artery disease or peripheral vascular disease, left ventricular hypertrophy, and dialysis duration >1 year, should have repeat cardiovascular assessment every 1-2 years. Transplant candidates and recipients should have individualized treatment for CVD and risk factors such as hypertension, diabetes, hyperlipidemia, and obesity. Special consideration should be given for statin therapy, as its use is associated with decreased cardiovascular death in dialysis and transplant patients. Prospective randomized, controlled trials are needed to determine the optimal approach to diagnosis and treat CVD in the transplant candidate and recipient population.

Crescentic Glomerulonephritis with Anti-GBM and p-ANCA Antibodies.

We are presenting a case of renal failure with anti-GBM and p-ANCA antibodies positive. Patients with dual antibodies are considered to be a vasculitis-variant of anti-GBM antibody nephritis. These patients may have atypical presentation and it may delay diagnosis and treatment. Recurrence rate is higher in these patients. We reviewed the literature of cases and studies on cresenteric glomerulonephritis with anti-GBM and p-ANCA positive patients. We recommend that patients suspected with pulmonary-renal syndrome should be checked for anti-GBM and p-ANCA antibodies, should undergo renal biopsy and should should have close long term follow up to watch for recurrence.

Calreticulin transacetylase catalyzed activation of rat tracheal smooth muscle cell nitric oxide synthase by acetoxycoumarins.

The Transacetylase function of Calreticulin (CR) catalyzing the transfer of acetyl groups from acetoxycoumarins (AC) to certain proteins was identified for the first time in our laboratory. Protein acetyltransferase action of CR was termed Calreticulin Transacetylase (CRTAase). In the present work, CRTAase of rat tracheal smooth muscle cells (TSMC) was characterized with respect to the specificity for various AC and its role in the activation of nitric oxide synthase (NOS). 7,8-Diacetoxy-4-methylcoumarin (DAMC), a model AC, when incubated with TSMC along with L-arginine caused profound activation of NOS as compared to that with L-arginine alone. Further, the inclusion of N-omega-nitro-L-arginine methyl ester (L-NAME) along with DAMC resulted in the reduction of NO levels of TSMC to that of control, there by confirming the activation of TSMC NOS. Also, several AC were found to activate TSMC NOS in tune with their specificities to CRTAase. The results presented in this paper bear evidence for the activation of TSMC NOS by AC and their effectiveness to enhance NO of airway cells may be expected to find useful applications in respiratory diseases.

Acetoxy drug: protein transacetylase catalyzed activation of human platelet nitric oxide synthase by polyphenolic peracetates.

An enhanced intracellular level of Nitric oxide (NO) is essential to ameliorate several pathological conditions of heart and vasculature necessitating the activation of NOS. We have projected in this report the acetylation of eNOS by polyphenolic peracetates (PA) catalyzed by the novel enzyme acetoxy drug: protein transacetylase (TAase) discovered in our laboratory as an unambiguous way of activating NOS which results in the manifestation of physiological action. The human platelet was chosen as the experimental system in order to validate the aforementioned proposition. PA caused profound irreversible activation of platelet NADPH cytochrome c reductase mediated by TAase. The convincing biochemical evidences are presented to show that PA could cause acetylation of the reductase domain of NOS leading to the activation of eNOS in tune with their specificities to platelet TAase. As a result, the enhanced level of NO due to activation of platelet eNOS by PA was found to inhibit the ADP-induced platelet aggregation. The present studies highlight for the first time the role of PA as the novel potent agent for enhancing the intracellular NO levels.