Polymerase chain reaction-based DNA analysis for gender identification of antemortem and postmortem human skin

Gender identification is an important investigative tool as it can be used to assess rapid information in forensic cases involving missing persons, intersex problems, mass disaster and in crime scenes. The aim of the present work was to assess the reliability of the skin as a tissue sample whether fresh or putrified for gender determination and to validate an application of gene print sex determination system. Forty skin samples were used [15 obtained from cadavers, and 25 from discarded surgically resected skin]. DNA extraction was done using two methods [crude and column] with two different kits, [Puregene cell, tissue kit and GFX purification kit]. Amplification of the single copy X-Y homologous amelogenin gene using PCR technology was followed. The results showed that DNA extracted by the column method was of high quality with no gross contaminants as compared to the crude method. The success rate of the amelogenin amplification was 100% in all skin samples [antemortem and postmortem]. It enabled gender identification from as low as 100 mg skin sample with low cost and less complicated technique. In conclusion the amelogenin gene sex determination system is a highly discriminating and reliable method and skin can be used as a good source for DNA extraction used for identification purposes

Study of some autoantibodies in cases of lymphoproliferatve disease

lymphoproliferative disorders are usually complicated by autoimmune disorders, the aim of this work is to study the autoimmune profile in some cases of lymphoproliferative disease. Material and Sixty cases of B-lymphoproliferative disorders, forty-five cases of non-Hodgkin lymphoma [NHL] and fifteen chronic lymphocytic leukemia [CLL] were included in this study. The autoimmune profile including; anti-nuclear antibodies, anti-ds-DNA anti-cardiolipin antibodies, both IgG and IgM were evaluated. 42% percent of studied cases displayed one or more autoimmune marker positivity. A total thirteen out of sixty [22%] were postive for anti-nuclear antibodies; three out fifteen CLL cases [20%] and ten out of forty-five non-Hodgkin lymphoma cases [22%]. Two out of fifteen [13%] CLL cases and eight out of forty-five [18%] NHL cases were positive for anti-ds-DNA antibodies. Only thirty-eight cases were tested for anti-cardiolipin antibodies; three case were acL IgG positive and nine were acL IgM positive. Although there was a high percent of positive autoimmune marker, SLE was diagnosed clinically in only of the NHL cases [2.2%]. HCV seropositivity was tested to exclude HCV as a contributing factor for autoantibodies development. Causes of autoimmune phenomena in lymphoproliferative conditions are heterogeneous and the association may not necessarily causal; certain individuals may be genetically predisposed to develop both disorders i.e. that genetic factors disturbing the regulation of the immune system may predispose both to lymphoid neoplasms and to autoimmune disease

Study of the mutations in the NAD[p]H: quinone oxidoreductase in cases of chronic myeloid leukemia

NAD [p] H: quinone oxidoreductase NQO1 originally called DT-diaphorase is an enzyme that is able to detoxify a number of natural and synthetic compounds, including quinones and their derivatives. It is induced by synthetic antioxidants and cruciferous vegetables and protects cells against oxidative stress. The human NQO1gene is located on chromosome 16q22.1, a single nucleotide polymorphism [cytosine to thymine, [CT]] at position 609 in the NQO1 gene has been identified in a human colon cancer cell line with very low NQO1 activity. This variant produces a proline-to-serine substitution that inactivates the enzyme. People who are homozygous for the variant allele completely lack NQO1 activity, while heterozygote have low to intermediate activity compared with people with the wild type. The incidence of the polymorphism varies widely by race, and associations have been made between the presence of variant alleles and lung and urological cancers. NQO1 mutation had been identified as risk factor in AML and ALL but no avilable data in chronic myeloid leukemia. Our study included 21 chronic myeloid patients and 10 apparently healthy persons using PCR-RFLP technique to detect the NQO1 nutation. Among the cases 11[52.4%] were positive for the mutation compared to 1[10%] of control. A higher incidence of the mutation was noticed among patients in crisis stage. Conclusion from our data we noticed a high incidence of NQO1 mutation in cases of CML which may suggest an important role of this gene in the etiology and pathogenesis of chronic myeloid leukemia