No abstract

No abstract

No abstract

Hypoxia is an integral part of tumorigenesis and contributes extensively to the neoplastic phenotype including drug resistance and genomic instability. It has also been reported that hypoxia results in global demethylation. Because a majority of the cytosine-phosphate-guanine (CpG) islands are found within the repeat elements of DNA, and are usually methylated under normoxic conditions, we suggested that retrotransposable Alu or short interspersed nuclear elements (SINEs) which show altered methylation and associated changes of gene expression during hypoxia, could be associated with genomic instability. U87MG glioblastoma cells were cultured in 0.1% O2 for 6 weeks and compared with cells cultured in 21% O2 for the same duration. Real-time PCR analysis showed a significant increase in SINE and reverse transcriptase coding long interspersed nuclear element (LINE) transcripts during hypoxia. Sequencing of bisulphite treated DNA as well as the Combined Bisulfite Restriction Analysis (COBRA) assay showed that the SINE loci studied underwent significant hypomethylation though there was patchy hypermethylation at a few sites. The inter-alu PCR profile of DNA from cells cultured under 6-week hypoxia, its 4-week revert back to normoxia and 6-week normoxia showed several changes in the band pattern indicating increased alu mediated genomic alteration. Our results show that aberrant methylation leading to increased transcription of SINE and reverse transcriptase associated LINE elements could lead to increased genomic instability in hypoxia. This might be a cause of genetic heterogeneity in tumours especially in variegated hypoxic environment and lead to a development of foci of more aggressive tumour cells.

No abstract

NAD(P)H quinone oxidoreductase 1 (NQO1) catalyzes reactions having a cyto-protective effect against redox cycling and oxidative stress. A single base polymorphism (C/T) at nucleotide 609 of the NQO1 gene impairs the stability and function of its protein. Its role in the development of diabetic nephropathy (DN) has not been deciphered. Therefore, this study aimed to evaluate the association of NQO1*2 (rs1800566) polymorphism with plasma NQO1 levels and DN. This study screened 600 participants including healthy controls (HC), type 2 diabetes mellitus without complications (T2DM) and diabetic nephropathy (DN): 200 each for studying NQO1*2 gene polymorphism using the PCR-RFLP. Plasma NQO1 levels were measured by ELISA. Analysis of variance and logistic regression were used to evaluate the association of NQO1 polymorphism with plasma NQO1 levels and DN. The allelic frequencies of NQO1*1/NQO1*2 were 0.88/0.12 in HC, 0.765/0.235 in T2DM and 0.65/0.35 in DN. Carriers of the NQO1*2 allele had significantly lower plasma NQO1 levels (p<0.05) and revealed higher risk towards the development of DN (OR=1.717, p=0.010). NQO1*2 SNP is a functional polymorphism having a significant effect on NQO1 levels. Our results indicate that NQO1*2 genotype may increase susceptibility to DN in north Indian subjects with T2DM.

New variety of pH-sensitive hydrogels, having macroporous interior with honey-comb type architecture and continuous skin at the surface, have been developed by single step aqueous copolymerization of acrylic acid (AAc) and N-[3-(dimethylamino)propyl]-methacrylamide (DMAPMA) in different feed ratios at 41 AE 1 C. Interlocked nanogels of $ 300 nm were identified as the building blocks in all of these cylindrical poly(-AAc-co-DMAPMA) matrices (PDMAAc). The gels showed good compressive strength even at a swelling as high as 4330% (mass) at pH 7.0. Morphology of McCoy fibroblast cell line remained unchanged in direct contact with different PDMAAc gels, and cell viability (AESD) was recorded to be in the range of 105 (AE3)% to 87 (AE8)% after 72 h. Bovine serum albumin (BSA) loaded gels were prepared by means of equilibrium partitioning. Loading efficiency of PDMAAc gels has been found to be in the range of 210-277 mg/g dry gel. BSA release from PDMAAc gels at 37 C has been found to follow nonFickian diffusion mechanism in simulated gastric juice (pH 1.2), and Case II transport in simulated intestinal juice (pH 7.4). In vitro study showed that the gels are capable of retaining >95% of the loaded BSA in gastric medium through average gastric emptying period. Again, $ 56% BSA release was recorded in 24 h at pH 7.4, indicating prolonged BSA diffusion in intestinal condition. Constant rate of BSA diffusion was reflected from the release profiles at both the pH. Diffusion exponents also supported the same and indicated for absolute zero-order kinetics at pH 7.4.

This study highlights the fact that establishing primary cultures from ascitic fluid or solid tumor tissue may help us to understand the molecular profile of the cancer cells, which allow us to select the best chemotherapeutic agent for ovarian cancer patients and thus take a step toward patient-tailored therapy so that patients are not exposed to drugs to which they are not likely to respond.