Genomic and serum tumor markers in Egyptian females with and without family cancer history

OBJECTIVE: Multiple variables affect the probability of development of cancer. The present study aimed to screen Egyptian females for early prognostic cancer markers such as carcinoembryonic antigen (CEA), the soluble form of transmembrane mucin protein (CA15-3), MUC1 and important sex hormones (Progesteron, Oestrogen, and Prolactin) and three germline BRCA1/2 founder mutations. PATIENTS AND METHODS: Forty-five DNA samples were screened for 185delAG and 5382insC in the BRCA1 and 6174delT in the BRCA2 genes using polymerase chain reaction (PCR)-directed mutagenesis. Each sample of the 185delAG and the 6174delT mutations was confirmed using Restriction Fragment Length Polymorphism (RFLP) analysis. Nine suspected PCR products of 185delAG and the forty-five amplicons of 6174delT mutations were further confirmed using Sanger sequencing. Sex hormones (Progesteron, Oestrogen, and Prolactin) and cancer antigens (CA 15-3 and CEA) concentrations were quantitatively determined in serum samples using ELISA. RESULTS: We found significant associations only for oestrogen (p-value=0.036), while non-significant (p-value= 0.123) hyperprolactinemia with cancer history. But none of the individuals carried the BRCA1/2 studied mutations while new variants were detected; (delA) in position 93865, deletion (delA) or substitution of A by G (A/G) in position 93858 and (insA) in position 93844 with frequency of 50%, 50%, 25% and 25%, respectively, in subjects with cancer history. CONCLUSIONS: The serum level of oestrogen could be a useful non-invasive cancer marker while significant association of hyperprolactinemia and the new BRCA1/2 variants with cancer needs extra study

Diversity in growth and expression pattern of PoHKT1 and PoVHA transporter genes under NaCl stress in Portulaca oleracea taxa

Plant growth and the expression of two transporter genes; PoHKT1 and PoVHA transcripts in root and shoot tissues were studied under salt stress of three Portulaca oleracea s.l. taxa. The study showed no significant differences in ratios between root lengths in saline and non-saline treatments of the three taxa, which was correlated with a clear down-regulation of the PoHKT1 transcripts in the root after 150mM NaCl. All measured growth parameters except root length increased in P. oleraceae, decreased in P. granulatostellulata and remain unchanged after 100mM NaCl in P. nitida compared to control under saline conditions. The result was consistent with the type of taxon which had significant effect on the shoot length, number of leaves and dry weight (P< 0.05). All measured growth parameters except root length showed a significant negative correlation with the shoot fold change of PoHKT1 transcripts (r = -0.607, -0.693 and -0.657 respectively). The regulation of PoVHA in root and shoot tissues in the three taxa are significantly different. Under salt stress, both decreased uptake of Na+ into the cytosol by decreasing the expression of PoHKT1 and increased vascular compartmentalization ability of Na+ by inducing the expression of PoVHA seem to work more efficiently in P. oleraceae and P. nitida than in P. granulato-stellulata

Validation of New ELISA Technique for Detection of Aflatoxin B1 Contamination in Food Products versus HPLC and VICAM

Toxin-contaminated foods and beverages are a major source of illness, may cause death, and have a significant negative economic impact worldwide. Aflatoxin B1 (AFB1) is a potent toxin that may induce cancer after chronic low-level exposure. This study developed a quantitative recombinant AflR gene antiserum ELISA technique for aflatoxin B1 detection in contaminated food products. Aflatoxin B1 residuals from 36 food samples were analyzed with HPLC and VICAM. DNA was extracted from aflatoxin-contaminated samples and the AflR gene amplified using PCR. PCR products were purified and ligated into the pGEM-T vector. Recombinant plasmids were sequenced and transformed into competent E. coli (BL21). Molecular size and B-cell epitope prediction for the recombinant protein were assessed. The purified protein was used to induce the production of IgG antibodies in rabbits. Serum IgG was purified and labeled with alkaline phosphatase. Finally, indirect-ELISA was used to test the effectiveness of polyclonal antibodies for detection of aflatoxin B1 in food samples