Browsing by Author "Ravichandran, M"
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Item Allele-specific PCR for a cost-effective & time-efficient diagnostic screening of spinal muscular atrophy.(2012-01) Marini, M; Sasongko, T H; Watihayati, M S; Atif, A B; Hayati, F; Gunadi; Zabidi-Hussin, Z A M H; Ravichandran, M; Nishio, H; Zilfalil, B ABackground & objectives: Genetic diagnosis of spinal muscular atrophy (SMA) is complicated by the presence of SMN2 gene as majority of SMA patients show absence or deletion of SMN1 gene. PCR may amplify both the genes non selectively in presence of high amount of DNA. We evaluated whether allelespecific PCR for diagnostic screening of SMA is reliable in the presence of high amount of genomic DNA, which is commonly used when performing diagnostic screening using restriction enzymes. Methods: A total of 126 blood DNA samples were tested in amounts ranging 80-200 ng, referred for the genetic diagnosis of SMA using both conventional PCR-RFLP and allele-specific PCR. Results: The results from both methods showed agreement. Further, allele-specific PCR was found to be a time-efficient and cost-effective method. Interpretation & conclusions: Our study demonstrated the accuracy of our allele-specific PCR and the results were comparable compatible with that of PCR-RFLP, indicating its practical application in SMA diagnostic screening.Item Detection of Cryptosporidium parvum in HIV-infected patients in Malaysia using a molecular approach.(2008-05-21) Zaidah, A R; Chan, Y Y; Asma, H Siti; Abdullah, Shukri; Nurhaslindawati, A R; Salleh, Mustafa; Zeehaida, M; Lalitha, P; Mustafa, Mahiran; Ravichandran, MThis cross-sectional study determined the prevalence of cryptosporidiosis in HIV-infected patients using polymerase chain reaction (PCR). Stool specimens were collected from HIV infected patients who were admitted to Hospital Raja Perempuan Zainab II, Kota Bharu, Malaysia, for various indications from December 2004 to December 2005. A modified acid-fast stain was performed on the direct stool smears, then the stool specimens were further tested using nested PCR targeting the 18S rRNA gene of Cryptosporidium parvum, with a built-in internal control (IC). Out of 59 samples, 11 were positives. Nested PCR identified a total of nine samples (16%) compared to microscopy, which identified only three samples. All PCR negative results showed IC amplicons, suggesting that these samples were true negatives and were not due to inhibition of PCR. This study highlights the importance of molecular diagnosis in determining the true prevalence and epidemiology of C. parvum.Item Effect of Bacillus subtilis PB6, a natural probiotic on colon mucosal inflammation and plasma cytokines levels in inflammatory bowel disease.(2009-02-21) Selvam, R; Maheswari, P; Kavitha, P; Ravichandran, M; Sas, Benedikt; Ramchand, C NThe pathophysiology of inflammatory bowel disease (IBD) involves the production of diverse lipid mediators, namely eicosanoid, lysophospholipids, and platelet-activating factor, in which phospholipase A2 (PLA2) is the key enzyme. Thus, it has been postulated that control of lipid mediators production by inhibition of PLA2 would be useful for the treatment of IBD. This hypothesis has been tested in the present study by examining the therapeutic effect of a novel natural probitic Bacillus subtilis PB6 (ATCC- PTA 6737). B. subtilis PB6 is found to secrete surfactins (cyclic lipopeptides) which have anti-bacterial potential. These surfactins inhibit PLA2, a rate-limiting enzyme involved in the arachidonic acid associated inflammatory pathway and could downregulate the inflammatory response by regulating the eicosanoid and cytokine pathways. With this concept, an experimental animal trial has been conducted in a rat model of 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. The oral administration of PB6 suppresses the colitis as measured by mortality rate, changes in the weight gain, colon morphology and the levels of plasma cytokines. The animals treated orally with PB6 at 1.5 x 10(8) CFU/kg thrice daily from day 4 to 10 significantly improve gross pathology of the colon and regain the colon weight to normal (p < 0.05), compared to TNBS-induced positive control. The plasma levels of pro-inflammatory cytokines (TNF-alpha, 1L-1beta, IL-6 and IFN-gamma) are also significantly lowered (p < 0.05) and anti-inflammatory cytokine (IL-I0 and TGF-beta) significantly (p < 0.05) increased after the oral administration of PB6 on day 11. The present study supports the concept that PB6 inhibits PLA2 by the secreting surfactins. In a clinical investigation, it is found to be well tolerated by all the healthy volunteers.Item Status of heavy metals distribution in municipal solid waste in Tiruchirappalli City, India.(2012-07) Noorjahan, A; Dhanakumar, S; Mohanraj, R; Ravichandran, MUrbanization and rapid growth of population in India has led to drastic increase in municipal solid waste. Unscientific disposal of municipal solid waste is one of the main reasons attributed for environmental degradation. The present work concentrates on municipal solid waste management in Tiruchirappalli City which comprises of four zones namely Srirangam, Goldenrock, Araiyamangalam and Abishekapuram. This study also attempted to assess the physical composition, characteristics and the heavy metals content in municipal solid waste. It can be observed that the bio-degradable fraction of municipal solid waste is found to be 74 percent of the total solid waste generated from the city. Hence composting could be the best option for the treatment of municipal solid waste.