Molecular characterization of glutathione S-transferase, endothelial nitric oxide synthase and Vitamin D receptor genes in breast cancer cases

Enzymes of the Glutathione S-transferase system [GST] modulate the effects of exposure to several cytotoxic and genotoxic agents. Nitric oxide [NO] is constitutively synthesized in the endothelium by endothelial nitric oxide synthase [eNOS] and acts as a pleiotropic regulator involved in carcinogenesis. Vitamin D levels may influence breast cancer development. The vitamin D receptor [VDR] is a crucial mediator for the cellular effects of vitamin D and additionally interacts with other cell-signaling pathways that influence cancer development. To check for the association of polymorphisms of GST, eNOS3 and VDR genes with the susceptibility and severity of breast cancer in Egyptian cases. This work included 100 cases with breast cancer and 100 healthy individuals. The mean age of cases was 48.31 +/- 11.40 years. They included 100 females. DNA was amplified using PCR-RFLP for detection of polymorphisms related to eNOS3 and VDR, also DNA was amplified using PCR-SSP for detection of polymorphisms related to GST and calculating the odds ratios and their 95% confidence intervals. Total cases showed high significant frequency of eNOS3[-786] CC [P<0.05, OR=18.58] genotypes, GSTT1 [null] [OR = 2.68; CI 95%=1.51-4.75; p=0.001]. These were considered risk genotypes for disease susceptibility. On the other hand, total cases showed low significant frequency with homozygosity for eNOS3[-786] TT [P=0.01] and the GSTT1 gene was present in 42.0% of the cancers and in 66.0% of controls [OR = 0.37; CI 95%= 0.21-0.66; p=0.001]. These may be considered low risk genotypes. No significant difference in frequencies of null and present genotypes of GSTM1 and VDR FOKI in total cases compared to controls. Polymorphisms related to eNOS3[-786], GSTT1 and VDR FOKI genes may be considered genetic markers for BC among Egyptian cases. This may have potential impact on family counselling as well as future management plans

[Knowledge, attitudes and practices related to cutaneous leishmaniasis among attendants of primary health care centers in Qassim Region - Saudi Arabia]

To determine knowledge of/and attitudes towards the infection with cutaneous leishmaniasis and the related practices among attendants of primary care centers of Qassim region. A descriptive cross sectional study. Participants [398] were interviewed and completed a structured questionnaire focusing on disease knowledge, attitudes and preventive practices. Approximately 43.2% of the participants showed an insufficient level of knowledge about the disease. The lowest level of knowledge was about how the disease is transmitted or prevented. The most common symptoms reported were: a lesion [54.0%] and a scar [30.4%]. The infectious nature of the disease was known to 14.6% while 29.9% didn't know that the disease can be treated. The majority failed to link the disease spreads with vector bites [91.5%]. Most of participants [73.6%] did not recognized the biting time of the vector. Regression analysis ranked education, high income and presence or history of a case within the family the most significant determinants of knowledge variable. Participants showed insufficient knowledge regarding the disease transmission and the poor related protective practices against the transmitting vector. Educational interventions using different media are recommended

Prevalence of alpha-1-antitrypsin gene mutations in Saudi Arabia

alpha-1 antitrypsin [AAT] deficiency results from mutations of the protease inhibitor [PI]. The AAT gene is mapped on chromosome 14 and has been associated with chronic liver disease and chronic obstructive pulmonary disease [COPD]. To determine the frequency of AAT mutations on S and Z carrier alleles in healthy Saudi individuals from Qassim Province in Saudi Arabia. A total of 158 healthy, unrelated participants from Qassim Province were recruited. They were genotyped for the two AAT-deficiency alleles, PI*S and PI*Z, using polymerase chain reaction, with primers designed throughout to mediate site-directed mutagenesis. Of the 158 subjects, 11.39% were carriers for the S mutation [i.e., had the MS genotype], whereas 2.53% were carriers for the Z mutation [i.e., had the MZ genotype]. The SZ genotype was present in 3.8% of subjects, while the homozygous genotype SS was present in 1.9% of subjects. No subjects showed the ZZ mutant genotype. Accordingly, frequency of the mutant S and Z alleles of AAT gene was 9.49% and 3.19%, respectively. The results obtained showed a high prevalence of the AAT deficiency allele in the Saudi population. This probably warrants adoption of a screening program for at-risk individuals, so that they might initiate adequate prophylactic measures

Association of factor V Leiden mutation with deep vein thrombosis among Egyptian cases

Deep vein thrombosis [DVT] is a blood clot in a major vein, usually in the legs and/or pelvis. If part of the thrombus breaks off, it becomes an embolism, which can travel through the heart and block the arteries to the lungs. Factor V Leiden [FVL] is a common genetic risk factor for hereditary hypercoagulability disorder in several populations. The present study investigates the association of FVL mutation with DVT among Egyptian cases. The study included 44 cases [16 males and 28 females] with an age range of 20 to 80 years in addition to 211 healthy unrelated controls of matched age and sex. A multiplex allele-specific PCR amplification was conducted for assignment of FVL gene mutation [G1691 A]. Cases having the mutant allele A [AA and AG genotypes] were significantly higher than controls [38.6% vs. 18.5%; P < 0.05, OR= 2.78 and CI 95%, 1.380-5.589]. These results concluded that FVL mutation has a high frequency and positive association with the occurrence of deep vein thrombosis among Egyptian cases

Rapid sex determination using PCR technique compared to classic cytogenetics

Fetal sexual differentiation relies on the translation of chromosomal sex established at fertilization into gonadal sex and somatic sex as development proceeds. In cases where chromosomal, gonadal, and somatic sex are incongruent in human infants and children, rapid establishment of the diagnosis and implementation of medical and surgical management is of paramount importance, since the gender identity is so important to the psychological well-being throughout life. This work was done in order to test the value of PCR technique for rapid sex determination compared to classic cytogenetic technique. Subjects included 20, cases including 10 neonates with ambiguous genitalia, 2 adult females with delayed puberty and 8 adult males with infertility, in addition to 20 normal infants of both sexes as a control group. The diagnosis of sex was attempted through examination, cytogenetic study, ultrasonography, gonadal biopsy and hormonal analysis, in addition to PCR amplification for the detection of SRY and ATL1 gene loci on Y and X chromosomes respectively. Four neonates were diagnosed as partial testicular feminization showed both positive bands for the Y and X chromosomes and a karyogram of 46/XY. Three neonates were diagnosed as true hermaphrodites showed positive amplification for both Y and X chromosomes with a mosaic karyogram 46, XX/XY. Three neonates were diagnosed as cases of adrenogenital syndrome showed positive amplification of only the Xchromosome and had a karyogram of 46/XX. One of the two adult females was diagnosed as Turner syndrome showed positive amplification of the X chromosome and a karyogram of 45/XO; the other one was diagnosed as complete testicular feminization had a positive amplification of X and Y chromosomes and a karyogram of 46/XY. The 8 adult males with infertility showed a positive amplification of X and Y chromosome and a karyogram of 47/XXY [Klinefelter syndrome] in 7 cases and 46/XY gonadal dysgenesis in one case. We concluded that PCR as a simple, rapid and reliable technique can complement and also confirm cytogenetic studies in the diagnosis of sex in cases of sex chromosome disorders

IL-10 Implications in Psoriasis

Interleukin [IL]-10 is a pluripotent cytokine with effects on numerous cell populations, in particular circulating and resident immune cells as well as epithelial cells. With its potent immunoregulatory capacities, its main biological function seems to be the limitation and termination of inflammatory responses. Hence, its low level expression found in psoriasis may have pathophysiological relevance to this immune disease. Remarkably, the induction of IL-10 expression was found by conventional antipsoriatic therapies, supporting the hypothesis that it may be a key cytokine in psoriasis. Furthermore, the first use in clinical trials in patients with established psoriasis showed that it had moderate antipsoriatic effects and was well tolerated. Moreover, long-term application in psoriatic patients in remission showed that it decreases the incidence of relapse and prolongs the disease free interval. The IL-10 antipsoriatic activity is suggested to be due to the effects on different cell populations, including antigen presenting cells and T-cells [type 1/type 2 balance shift], but not through direct effects on keratinocytes. In conclusion, IL-10 seems to have major clinical and therapeutic implications in psoriasis. Further multicenter, placebo-controlled, double blind trials are required to be an established antipsoriatic therapy. We can come to the conclusion that IL-10 genetic polymorphism and expression is potentially a key immune marker in psoriasis

Diagnosis of sex chromosome disorders and prenatal diagnosis of Down Syndrome using interphase Fluorescent In-Situ Hyperidization Technique

Background: Thousands of infants are born each year with chromosomal abnormalities that severely impact physical and mental development. Among common genetic disorders are Down syndrome [trisomy 21] and sex chromosomal disorders

Objectives: Evaluation of guidelines used for prenatal diagnosis of Down syndrome [DS] as well as sex chromosomal disorders including interphase Fluorescent In Situ Hyperidization [FISH] technique

Methods: Enrolled cases were among those presenting to Genetics and Neonatology Units, Mansoura and Ain-Shams University hospitals,[Egypt] during 2002 to 2004. These included: Groups 1 comprised fifty pregnant women presenting for genetic counseling. They were subjected to complete history analysis, ultrasound examination in addition to triple screening test [for alpha feto protein [AFP], human chorionic goandotrophin [HCG] and unconjugated esteriol [E2]. Results were confirmed by doing routine karyogram on cultured amniotic fluid. Groups 2 comprised suspected cases with sex chromosomal disorders including neonates with ambiguous genitalia [64 cases] and adults with primary amenorrhea [69 cases] or infertility [38 cases]. They were subjected to a diagnostic workup including

Results: Among the pregnant women group, seven were found to be at a high risk of having DS fetuses including 3 cases with a history of affected off-springs, 2 cases with age above 35 years, and 2 cases with high triple test. Only one case had positive trisomy 21 on interphase FISH confirmed by karyogram on cultured amniotic cells. The other 6 ladies had normal FISH confirmed by karyograms. Regarding the other group, 5 cases out of the 9 females were proved to be feminized males, one proved mosaic turner, one proved mixed gonadal dysgenesis and 2 normal females. On the other hand one out of three males were proved to be verilized female while the other one was a male with incomplete testicular feminization and the last one was a male with infertility diagnosed as Klinefelter syndrome at the age of 26 years

Conclusion: Interphase FISH is a rapid, accurate and very sensitive method in sex chromosom and autosomal abnormalities. It adds to the diagnostic utility of routine cytogenetics and its use on interphase nuclei overcomes the difficulty of conventional cytogenetics. It could be used in the prenatal diagnosis of DS in addition to ultrasonography, and triple test

Diagnosis of sex chromosome disorders and prenatal diagnosis of Down syndrome using interphase fluorescent in-situ hyperidization technique

Thousands of infants are born each year with chromosomal abnormalities that severely impact physical and mental development. Among common genetic disorders are Down syndrome [trisomy 21] and sex chromosomal disorders. Evaluation of guidelines used for prenatal diagnosis of Down syndrome [DS] as well as sex chromosomal disorders including interphase Fluorescent In Situ Hyperidization [FISH] technique. Enrolled cases were among those presenting to Genetics and Neonatology Units, Mansoura and Ain-Shams University hospitals,[Egypt] during 2002 to 2004. These included: Groups 1 comprised fifty pregnant women presenting for genetic counseling. They were subjected to complete history analysis, ultrasound examination in addition to triple screening test [for alpha feto protein [AFP], human chorionic goandotrophin [HCG] and unconjugated esteriol [E2]. Results were confirmed by doing routine karyogram on cultured amniotic fluid. Groups 2 comprised suspected cases with sex chromosomal disorders including neonates with ambiguous genitalia [64 cases] and adults with primary amenorrhea [69 cases] or infertility [38 cases]. They were subjected to a diagnostic workup including. Among the pregnant women group, seven were found to be at a high risk of having DS fetuses including 3 cases with a history of affected off-springs, 2 cases with age above 35 years, and 2 cases with high triple test. Only one case had positive trisomy 21 on interphase FISH confirmed by karyogram on cultured amniotic cells. The other 6 ladies had normal FISH confirmed by karyograms. Regarding the other group, 5 cases out of the 9 females were proved to be feminized males, one proved mosaic turner, one proved mixed gonadal dysgenesis and 2 normal females. On the other hand one out of three males were proved to be verilized female while the other one was a male with incomplete testicular feminization and the last one was a male with infertility diagnosed as Klinefelter syndrome at the age of 26 years. Interphase FISH is a rapid, accurate and very sensitive method in sex chromosom and autosomal abnormalities. It adds to the diagnostic utility of routine cytogenetics and its use on interphase nuclei overcomes the difficulty of conventional cytogenetics. It could be used in the prenatal diagnosis of DS in addition to ultrasonography, and triple test

Molecular diagnosis of duchennemuscular dystrophy by multiplexpolymerase chain reaction

Duchenne or Becker muscular dystrophy [DMD/BMD] is one of the most common X-linked lethal genetic diseases with a worldwide frequency of one in 3500 live male births. The Dmd locus is, the largest human gene known, span 2.4 Mb in Xp21. It consists of 79 exons that encode a14 Kb transcript. We have investigated the frequency of deletion in the Dmd gene in 37 unrelated Duchenne muscular dystrophy [DMD] Egyptian patients. All patients were screened for 21 exonic deletion which pro-posed by Multicenter Study Group. The screening of deletion was done using single strand conformation polymorphism [SSCP] after multiplex PCR. The most common exonic deletion in our study are exon 48 [46 %],exons-19, 45 [37.8 % of each] followed by exons-43 [ 29.7 %], PM, 44, 47,and 50 [27 %]. The overall deletion in our study about 83.8% in DMD. Deletions were clustered at two spots: 76 % at the hot spot in the distal region of the gene and 24 % at the hot spot in the proximal region of the gene. The multiplex PCR is simple, reliable and rapid technique for detection of carrier in families with DMD