Genetic Screening of Selected Disease-Causing Mutations in Glutaryl-CoA Dehydrogenase Gene among Indian Patients with Glutaric Aciduria Type I.

Glutaric aciduria type I (GA-I) is an organic aciduria caused by glutaryl-CoA dehydrogenase (GCDH) deficiency. There are limited studies on GA-I from India. A total of 48 Indian GA-I patients were screened for selected disease-causing mutations such as R402W, A421V, A293T, R227P, and V400M using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Among these patients, 9 (18.8%) had R402W mutation, and none had A421V, A293T, R227P, or V400M mutation. One low excretor mutation (P286S) and several novel mutations (I152M, Q144P, and E414X) were also found in this study. We conclude that among selected mutations, R402W is the most common mutation found among Indian GA-I patients.

Development of SCAR marker associated with downy mildew disease resistance in pearl millet (Pennisetum glaucum L.).

Pearl Millet is an important crop coarse grain cereal crop in the semi arid tropics which is extremely susceptible to oomycete plant pathogen Sclerospora graminicola causing downy mildew (DM) disease. The aim of the current study is to breed resistance against downy mildew disease into high yielding cultivars of pearl millet. Hence, in the present work a sequence characterized amplified region (SCAR) marker was developed as a molecular screening tool to identify DM resistance source and presented here. Of the 27 inter simple sequence repeats (ISSR) decamer primers used to identify polymorphism amongst pearl millet genotypes ICMR-01007 (P1) and ICMR-01004 (P2) and their populations (F1 and F2), only one primer pair ISSR-22 produced polymorphic bands on ICMR-01004 producing 1.4 kb size. The PCR amplification of 1.4 kb band was found tightly linked to the resistant line of ICMR-01004 and also in F2 segregation population was in the ratio 3:1. This band was cloned, sequenced and candidate SCAR primer (SCAR ISSR 863 ) was designed. Segregant analysis of their F2 progeny revealed that the SCAR ISSR 863 marker was linked to downy mildew resistance linkage group (χ(2) 3:1 = 0.86, P = 0.22) with a genetic distance of 0.72 cM. This SCAR marker was further validated using diverse pearl millet lines of India and Africa. Results indicated that the SCAR ISSR 863 band was amplified in all the seven resistant lines and were absent in five susceptible lines. The confirmation of the ISSR-derived SCAR marker in different genetic backgrounds of pearl millet lines suggests that this marker can be exploited for DM resistance screening in pearl millet breeding programs.

Cytotoxic and pro-apoptotic effects of Abrus precatorius L. on human metastatic breast cancer cell line, MDA-MB-231.

Abrus precatorius is highly regarded as a universal panacea in the herbal medicine with diverse pharmacological activity spectra. This experimental study on the mechanism of the anticancer activity of A. precatorius leaf extracts, may offer new evidence for A. precatorius in the treatment of breast cancer in clinical practice. Cell death was determined by using MTT assay. Further analyses were carried out by doing DNA laddering, PARP cleavage, FACS, semi-quantitative RT-PCR and detection of cellular reactive oxygen species (ROS) by DCFDA assay. A. precatorius showed very striking inhibition on MDA-MB-231 cells. MTT assay showed more than 75 % inhibition of the cells and treated cells indicated visible laddering pattern with thick compact band. PARP cleavage produced 89 kDa cleavage product which was associated with apoptosis. Flow cytometer exhibited a sub-G0/G1 peak as an indicative of apoptosis. mRNA expression level of apoptosis-related genes p21 and p53 was markedly increased in cells treated with the extract as compared to control. The up-regulation of p21 and p53 may be the molecular mechanisms by which A. precatorius extract which induces apoptosis. An increase in the concentration of A. precatorius extract does not generate ROS, instead it reduces ROS formation in MDA-MB-231 cells, as evident from the shift in fluorescence below untreated control. This is the first report showing that A. precatorius leaf extract exhibits a growth inhibitory effect by induction of apoptosis in MDA-MB-231 cells. Our results contribute towards validation of the A. precatorius extract as a potentially effective chemopreventive or therapeutic agent against breast cancer.