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

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

Characterization of angiotensin converting enzyme gene polymorphism and risk of different heart diseases

The present work aims to test the association of angiotensin converting enzyme [ACE] gene insertion/ deletion [I/D] polymorphism in patients with myocardial infarction [MI]. The study comprised 79 Egyptian cases with MI. Their mean age was 54.4 +/- 9.9 years including 60 [75%] males and 19 [24.1%] females, 23 [29.1%] were smokers, 21 [26.6%] had a positive family history of MI, 25 cases [31.6%] were diabetic, 16 cases [20.3%] were hyperlipidemic. For comparison, 238 healthy subjects of nearly matched age and sex, with no history of any cardiac diseases were taken as a control group. For all subjects, DNA testing for ACE gene I/D polymorphism was done using PCR amplification to detect both D and I alleles followed by a second run PCR specific for the I allele for cases typed as DD in the first run. Cases had higher frequency of DD [29.1%] and ID [62%] than II [8.9%] genotype with a higher frequency of D allele than I allele [64.4% vs 33.6%]. Compared to controls, cases had significantly higher frequency of ID genotype [62% versus 47.5%, P < 0.05]. Cases with low risk factors had a higher frequency of ID genotype compared to controls [66.7% vs 47.5%, P = 0.002]. The same was, also, found in the high risk group but with a lower level of significance [63.6% vs 47.5%, P = 0.041]. ACE gene polymorphism is probably a risk factor for ischemic heart disease among Egyptian cases particularly if integrated with other environmental and genetic risk factors

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

Plasma endothelin-1 and nitric oxide in children with insulin-dependent diabetes mellitus

Dysfunction of vascular endothelium is considered an early step in the development of diabetic complications. To assess plasma endothelin-1 [ET-1] and nitric oxide [NO] levels in children with insulin-dependent diabetes mellitus [IDDM] and their relation to the degree of metabolic control and disease duration. Plasma ET-1 and NO levels were assessed-by enzyme immunoassay-in 34 children with IDDM and compared to 17 healthy controls of matched age and sex. Diabetic patients had higher plasma ET-1 levels compared to controls [median [IQR]=5.9 [4.9-39.2] Vs 4.9 [4.4-6.1] pg/mI, P=0.02]. ET-1 levels were higher in patients with poor and moderate metabolic control when compared to those with ideal control [p=0.004 and 0.001; respectively]. ET-1 levels were positively correlated with NO levels [r=0.48, p=0.004]; HblAc level [r=0.57, P=0,001]; and disease duration [r=0.39, p=0.02]. Although, plasma NO levels in diabetic patients were not significantly different from controls [median [IQR]=24.6 [21.9-30.2] Vs 22.0 [21.0-26.5] umol/L, P 0.09]; NO levels were significantly higher in patients with poor metabolic control when compared to those with ideal control [p<0.001]. In children with IDDM, poor metabolic control and increased disease duration are associated with increased ET-1 production, which may be related to future diabetic complications. The elevated plasma NO levels in poorly controlled patients might mean a compensatory protective response towards increased ET-1 production

Bone mineral density and serum calcium hotheostasis in Egyptian children and adolescents with insulin-dependent diabetes mellitus

Although osteopenia is often reported as a complication of insulin-dependent-diabetes mellitus [IDDM], studies of bone mineral density [BMD] and serum calcium homeostasis in IDDM have yielded conflicting results. To determine BMD and serum calcium homeostasis in children and adolescents with IDDM and evaluate its relationship to metabolic control and disease duration. BMD was measured by dual-energy X-ray absorptiometry [DXA] in the lumbar spine [L2-L4] in 38 patients with IDDM [14 males, 24 females; aged 4-15 years; duration of diabetes 1.5-10 years] and their values were compared to those of 352 healthy Egyptian children and adolescents-[195 boys and 157 girls, aged 1-15 years]. In addition, glycosylated hemoglobin [HbA1c], serum calcium, phosphorus, 25 [OH] D3, and intact parathyroid hormone [iPTH] were measured in the diabetic patients and in 12 healthy controls of matched age and sex. As compared to normal BMD of Egyptian children and adolescents, nine patients [23.7%] had severe osteopenia [Z-score-2 SD or more negative] and 5 patients [13.1%] had mild osteopenia [Z-score:-1 to<-2 SD]; 24 [63.2%] patients had normal BMD. A negative correlation was found between BMD Z-score and disease duration [r-0.44, p=0,01], No relationship was found between BMD Z-score and metabolic control [HbA1c] or calcium homeostatic parameters. Diabetic patients had significantly lower iPTH compared to controls. On the other hand, serum calcium, phosphorus, and 25 [OH] D3 were not significantly different in IDDM patients compared to controls. IDDM in Egyptian children and adolescents was associated with osteopenia, which is significantly related to disease duration but not the degree of metabolic control. Calcium homeostasis was within normal in IDDM. iPTH was significantly lower in diabetic patients compared to controls. Periodic assessment of BMD in diabetic children and adolescents is mandatory