Development of low-density oligonucleotide microarrays for detecting mutations causing wilson’s disease.

Background & objectives: Wilson’s disease (WD) is an autosomal recessive disorder caused by mutations in ATP7B, a copper transporter gene, leading to hepatic and neuropsychiatric manifestations due to copper accumulation. If diagnosed early, WD patients can be managed by medicines reducing morbidity and mortality. Diagnosis of this disease requires a combination of tests and at times is inconclusive due to overlap of the symptoms with other disorders. Genetic testing is the preferred alternative in such cases particularly for individuals with a family history. Use of DNA microarray for detecting mutations in ATP7B gene is gaining popularity because of the advantages it offers in terms of throughput and sensitivity. This study attempts to establish the quality analysis procedures for microarray based diagnosis of Wilson’s disease. Methods: A home-made microarrayer was used to print oligonucleotide based low-density microarrays for addressing 62 mutations causing Wilson’s disease reported from Indian population. Inter- and intra- array comparisons were used to study quality of the arrays. The arrays were validated by using mutant samples generated by site directed mutagenesis. Results: The hybridization reaction were found to be consistent across the surface of a given microarray. Our results have shown that 52 °C post-hybridization wash yields better reproducibility across experiments compared to 42 °C. Our arrays have shown > 80 per cent sensitivity in detecting these 62 mutations. Interpretation & conclusions: The present results demonstrate the design and evaluation of a low-density microarray for the detection of 62 mutations in ATP7B gene, and show that a microarray based approach can be cost-effective for detecting a large number of mutations simultaneously. This study also provides information on some of the important parameters required for microarray based diagnosis of genetic disorders.

Phytochemistry, traditional uses and cancer chemopreventive activity of Amla (Phyllanthus emblica): The Sustainer.

Ayurveda, which is the oldest health system in the world, appreciates and uses amla to treat a host of diseases and promote positive health. Amla [Emblica officinalis, or emblic myrobalan], is called amalaki in Sanskrit. It is extensively used as a rejuvenator in ayurveda. It is also used widely in combination with other two [chebulic and belleric] myrobalans [fruit-bearing plant species] as triphala. Amla is indeed, the key ingredient in the popular ayurvedic recipe, Chyavanaprasha. More than anything, it may be called as "King of Rasayana" [rejuvenation], owing to its multiple health benefits. Phyllanthus emblica or Indian gooseberry (Amla) possesses a vastethnomedical history and represents a phytochemical reservoir of heuristic medicinal value. It is one of the oldest oriental medicines mentioned in Ayurveda as potential remedy for various ailments. The fruit is rich in quercetin, phyllaemblic compounds, gallic acid, tannins, flavonoids, pectin and vitamin C and also contains various polyphenolic compounds. A wide range of phytochemical components including terpenoids, alkaloids, flavonoids, and tannins have been shown to posses’ useful biological activities. Many pharmacological studies have demonstrated the ability of the fruit shows antioxidant, anticarcinogenic, antitumour, antigenotoxic, antiinflammatory activities, supporting its traditional uses. In this review, we have focused our interest on phytochemistry, traditional uses, cancer chemopreventive activity of Phyllanthus emblica both in vivo and in vitro. In view of its reported pharmacological properties and relative safety, P. emblica could be a source of therapeutically useful products.