C-6 Ceramide Induces p53 Dependent Apoptosis in Human Astrocytoma Grade4 (Glioblastoma Multiforme) Cells.

Ceramide is composed of sphingosine and a fatty acid found in large concentration within the cell membrane and often acts as a signaling molecule for various functions including programmed cell death. In the present investigation, we observed that C6-ceramide induces p53-dependent apoptosis and effectively killed the Astrocytoma grade4 (Glioblastoma Multiforme) HTB12 cell lines. Ceramide-induced cell death was confirmed by Trypan blue assay which showed about 65% cells dying from ceramide treatment. Apoptosis was confirmed by Caspase3 ELISA assay and DNA fragmentation assay. The p53 induction was confirmed by immunoblot studies. Since C6 Ceramide induces apoptosis in Glioblastoma cells, it may be employed in chemotherapeutic strategy to treat this highly malignant brain cancer.

Molecular Diagnosis of Helicobacter Pylori Strain by 16S rDNA PCR Amplification and Direct Sequencing.

AIM: Rapid detection of H.pylori strains by PCR-Sequencing. METHODS: 16S rDNA amplification by PCR from template genomic DNA, confirmation of amplicon size by agarose gel electrophoresis, sequencing of amplicons by automated sequencer, analysis of sequences by NCBI -BLAST software. RESULTS: The PCR -Sequencing and analysis of the sequence data by BLAST resulted in detection of the strain to be of H.pylori strain#26695. CONCLUSION: The pathogenicity of H.pylori depends on the strain of the bacteria, PCR-Sequencing and analysis of the sequence data by BLAST can be a very quick and useful diagnostic method of the pathogen.

Effects of 2-amino-4,6-dinitrotoluene on p53 tumor suppressor gene expression.

2-Amino-4,6-dinitrotoluene (2-Am-DNT) and its isomers are the most common metabolites of 2,4,6-trinitrotoluene (TNT). It is desirable to know the toxicity of this metabolite particularly because of its role in carcinogenicity and because it could contaminate drinking water. We used MCF-7 human breast cancer cells which have p53 tumor suppressor gene in wild type form in both the loci. Immunoblotting with p53 antibody showed enhanced p53 level in treated cells compared to untreated control cells. Similarly, p53 DNA-protein binding assays (gel-shift) showed accumulation of p53 protein in treated cells. This is the first report which shows p53 accumulation in 2-Am-DNT treated cells providing evidence of potential carcinogenic effects of 2-Am-DNT.