Maysaa Adil Hadi
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Extraction of Genomic DNA from Formalin Fixed - Paraffin Embedded Tissue for Forensic DNA Analysis: Influence of Proteinase K Maysaa Adil Hadi
Journal of Global Pharma Technology Volume 11 Issue 03
Publisher : Journal of Global Pharma Technology

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Abstract

A huge part of tissues preserved throughout the world used in diagnostic aim is formalin-fixed and paraffin embedded tissue (FFPET) which are a highly worthy supply for retrospective (genetic) application contain screening of mutations in cancer-critical genes in addition to detection of pathogens. The purpose of present study was to estimate the best method in the extraction of DNA from FFPET for utilizing in the forensic analysis by comparing three various extraction methods with and without proteinase K digestion and estimated the influence of these procedures of extraction in DNA quality, quantity and polymerase chain reaction (PCR) amplicon. In this study, the comparison was between three DNA extraction methods from five identically processed FFPET. The extraction methods, including DNA extraction kit, phenol: chloroform: isoamyl alcohol with and without proteinase K methods. Amplifiable DNA fragment size was assessed by amplification of exon 7 of CYP1A1 gene and the size of the fragment was (312 bp). Extractions by different methods used in this study were found suitable for amplification of fragment (312 bp) from all tissues used. The best amplification was after extraction without proteinase K and DNA extraction kit. In conclusion, amplification by PCR shows extremely affected by the method of extraction utilized. Pre-treatment of proteinase K was not a prerequisite for extractions of DNA from FFPET and the suitable extraction method was phenol: chloroform: isoamyl alcohol method without proteinase K as it was less time, less cost, yielding a good DNA quantity and quality.Keywords: Formalin Fixed-Paraffin Embedded Tissue, Forensic Analysis, Proteinase K.
Effect of Flax Seeds (Linum usitatissimum L.) Extract in Male Reproductive System of Albino Rat Treated with Cyproterone Acetate Maysaa Adil Hadi
Journal of Global Pharma Technology Volume 9 Issue 10
Publisher : Journal of Global Pharma Technology

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Abstract

The purpose of this report to assess the possible protective effects of 20%methanolic extract of flax seeds on cyproterone acetate mediated reproductive dysfunction of adult albino rats. Albino rats (35) place din a random manner into 7 groups, 5 rats in each. The normal control rat was the first group, while the second control group was given corn oil. Third group received cyproterone acetate (CPA) (10 mg/kg) as (positive control). Fourth and fifth groups were treated with 250&500mg/kg of flaxseeds extract respectively. The other protected sixth and seventh groups received CPA (10mg/kg) with flaxseeds extract 250 and 500 mg/kg respectively. All groups treated for 50 days orally by gavages tube. In CPA group, the outcome displayed significant reduction (P≤0.05) in the body weight, sperm parameters and FSH level, a significant raise (P≤0.05) in sperms abnormality and level of testosterone, and non-significant decline (P˃0.05) in reproductive organs and the accessory glands/ body weight ratios and LH level as compared to oil control. Treatment with flaxseeds 250&500 mg/kg caused decrease in the body weight but it considered as significant raise(P≤0.05)in comparison with CPA group, while group treated with flaxseeds extract 250 mg/kg make on-significant changes (P˃0.05) in each of reproductive organs and accessory gland/body weight ratios, LH level record non-significant decrease (P˃0.05) and significant decrease(P≤0.05) in testosterone level as compared to control D.W and CPA group whereas treatment with flaxseeds extract 500 mg/kg recorded significant increase in testes and epididymis/body weight ratio as compared to D.W, corn oil controls, and CPA groups. Furthermore, oral administration of the flaxseeds extract with CPA at two doses 250 & 500 mg/kg with CPA could reduce the negative effect of CPA in the body weight but it could not enhance the most of the sperm parameters and histological changes of both testes and epididymis caused by CPA. In conclusion, CPA (10mg/kg)induce degenerative changes in the reproductive organs of male rats and the 20%methanolic extract of flaxseeds were exhibit similar effect of CPA but at less extent in the male reproductive system.Keywords: Flax seeds (linum usitatissimum L.), Cyproterone Acetate, Reproductive Male system, Albino Rat.