Carrier detection in Duchenne muscular dystrophy using molecular methods.

Background & objectives: Duchenne and Becker muscular dystrophies are X-linked allelic disorders which are caused by mutations in the DMD gene. Carrier analysis in DMD is complicated due to the heterozygous nature of the X chromosome. Several techniques have been tried for carrier analysis in families where the mutation is identified including quantitative multiplex PCR (qmPCR), Southern blot, and now multiplex ligation-dependent probe amplification (MLPA). Linkage analysis is used in cases without identifiable mutations. The present study was undertaken to determine the status of probable carriers in families where the DMD deletion/duplication has been identified for the affected index cases. Methods: Carrier status was present in 150 probable carriers from 110 apparently unrelated families where the patients’ mutations were known. Of these 110 families, 100 were deletions, 9 duplications and 1 point mutation. Multiplex ligation-dependent probe amplification (MLPA) was used to assess the copy number changes and direct sequencing was used for the case with the point mutation. Results: Of the 150 cases, 49 were found to be carriers. Among the sporadic cases, it was observed that the rate of de novo mutations was very high (71%) as compared to the hereditary cases (29%), which was higher than the calculated rate (30%). It was observed that this difference was more apparent in deletion mutations than in duplications. Interpretation & conclusions: Identifying the DMD carrier rates in the families with unidentified deletions and duplications and where the causative mutation could be small insertions/deletions or point mutations could throw more light into this observation. MLPA was found to be useful in detecting copy number changes in DMD carriers and this could be the method of choice for DMD carrier analysis, when the mutation is detected in the affected child.

Hepatoprotective effect of ethanol extract from leaves of Phyllanthus emblica in rats

In this study, phytochemical extraction was carried out on the leaves of Phyllanthus emblica using ethanol as the organic solvent. The antioxidant activity of ethanol extract from Phyllanthus emblica was evaluated by various biochemical marker parameters, including super oxide dismutase [SOD - anion radical scavenging], catalase [CAT - hydrogen peroxide scavenging], glutathione peroxidase [GPX], glutathione reductase [GRD - reducing power], glutathione transferase [GTS] and thio-barbituric acid reactive substances [TBARS] measurement. Hepato cellular damage was induced by intraperitoneal administration of CCl[4] in male albino Sprague-Dawley strain rats. The reduced levels of SOD, CAT, GPX and GRD with CCl[4] treated animals attain normal level, after administration of ethanolic extract of Phyllanthus emblica. However, the activity of GTS was significantly higher in CCl[4] treated animals, which were brought down towards normal level in herb treated rats. The increased concentration of TBARS with CCl[4] treated rats attains normal level after administration of ethanolic extract of the same plant. An antioxidant property appears to be predominantly responsible for this hepato-protective action of the drug

Antimicrobial activity of extracts and isolates of the leaves of Plumeria alba Linn.

Disc diffusion method was employed to determine the effect of methanol, chloroform and acetone extracts of the dried leaves of Plumeria alba Linn. [Apocyitaceac] against fungi [Candida albicans] and bacteria [Bacillus subtilis, Proteus vulgaris Staphylococcus aureus. Pseudomonas aeruginosa, Streptococcus fecalis, Escherichio coil, Staphylococcus albus and Klebsiella pneumoniae]. The methanol and acetone extracts exhibited a prominent antimicrobial activity. The methanolic and acetone extracts were further fractionated by column chromatography to yield 2 pure isolates. The methanol pure component-2 [MPC[2]-50% Ethyl acetate:Methanol] and Acetone pure component-2 [APC[2]-70% Acetone:Ethyl acetate] have shown significant activity against all organisms used except Klebsiella