Rizk El-Baz

Gene polymorphisms of TNF-α−308 (G/A), IL-10−1082 (G/A), IL-6−174 (G/C) and IL-1Ra (VNTR) in Egyptian cases with type 1 diabetes mellitus

Background. Type 1 diabetes (T1D) is a genetically conditioned autoimmune disease in which cytokines play an important role. Objectives. To check for the association of polymorphisms of cytokine genes with type 1 diabetes. Subjects. This work included 50 cases with T1D and 98 healthy individuals from the Nile Delta region of Egypt. Cases included 20 males and 30 females with a median age of 25 and range of 15–50 years. Methods. DNA was amplified using PCR with sequence-specific primers for detection of polymorphisms related to tumor necrosis factor (TNF)-α− 308 (G/A), interleukin (IL)-10− 1082 (G/A), IL-6− 174 (G/C), and IL-1Ra (VNTR). Results. Cases with T1D showed significant higher frequency of genotypes of TNF-α− 308 AA (p < 0.001, odds ratio (OR) = 7.91), IL-6-17CC (p < 0.05, OR = 3.36) and IL-1Ra A1A1 (p < 0.05, OR = 3.68) with significant lower frequencies of TNF-α− 308 GA, and IL-1Ra A1A2 genotypes (p < 0.001 and < 0.05, respectively). They also showed significant higher frequency of TNF-α− 308 allele A (p < 0.05, OR = 2.0), IL-1Ra allele A1 (p < 0.05, OR = 2.98) with a significant lower frequency of TNF-α− 308 G allele and IL-1Ra A2 allele (p < 0.05). No significant difference was detected among cases in relation to IL-10− 1082 (G/A) genotypes or alleles nor in relation to age, sex, consanguinity or family history of the disease. Conclusions. Polymorphisms related to TNF-α and IL-1Ra genes may be considered genetic markers for T1D among Egyptians with a potential impact on family counseling and management.

Angiotensin-converting enzyme gene insertion/deletion polymorphism in Egyptian patients with myocardial infarction.

Introduction. This work aimed to test the association of the angiotensin-converting enzyme gene insertion/deletion (I/D) polymorphism with myocardial infarction. Subjects and methods. This study comprised 79 Egyptian myocardial infarction cases with a mean age of 54.4±9.9 years including 60 males and 19 females, plus 238 healthy unrelated individuals of nearly matched age and sex as a control group. For all subjects, DNA testing for the angiotensin-converting enzyme gene I/D polymorphism was done using PCR amplification for detection of both the D and I alleles followed by a second run PCR specific for the I allele for samples typed as DD in the first run. Results. Cases had a higher frequency of DD (29.1%) and ID (62.0%) genotypes 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 a significantly higher frequency of ID genotype (62.0% vs. 47.5%, p<0.05).This was more apparent among cases in the low risk group (p=0.002) than in the high risk group (p=0.041). Conclusion. The angiotensin-converting enzyme gene I/D polymorphism is probably a risk factor for ischaemic heart disease among Egyptian cases, particularly if integrated with other environmental and genetic risk factors.

Frequency of factor V Leiden mutation in Egyptian cases with myocardial infarction

Abstract Background: Acute myocardial infarction (MI) is death or necrosis of myocardial cells due to lack of blood supply. One of the causes may be thrombosis resulting from inherited resistance to activated protein C caused by the factor V Leiden (FVL) mutation. Objectives: To check for the presence of FVL mutation among Egyptian cases with MI compared to normal population controls. Subjects and methods: This study is a form of a prospective controlled study including 44 MI cases with an age ranging from 25 to 80 years and sex of 36 males (81·8%) and 8 females (18·2%). These cases were taken randomly from those admitted in the Intensive Care Units of Mansoura University Hospitals, Egypt. Of these cases, 10 cases (55·6%) were smokers, 7 cases (75·0%) had a positive family history of MI, 8 cases (18·18%) were diabetic and 20 cases (45·45%) were hyperlipidemic. For association and risk analysis, cases were compared to 211 healthy unrelated control subjects of matched age and sex. Factor V Leiden (G1691A) gene mutation was detected using a multiplex allele-specific PCR amplification. Results: Mutant A allele frequency of factor V Leiden was significantly higher in cases (30·68%) than in controls (10·19%) (p<0.0001, OR=3·9). Total cases showed significant higher frequency heterozygous mutant genotype GA (43·0%) compared to controls (16·6%), (p<0.0001, OR=4·45). Also total cases showed significant higher frequency of the homozygous mutant genotype AA (9·0%) compared to controls (1·9%), (p=0.0094, OR=8·19). On the other hand, no significant difference was found between cases subgroups related to age, sex, smoking, diabetes and hyperlipedemia. Conclusion: Frequency of factor V Leiden mutation among Egyptian cases with myocardial infarction is relatively high. So, families of affected subjects should be genotyped and counseled for proper prophylaxis.

Methylenetetrahydrofolate Reductase Gene Polymorphisms in Egyptian Women with Unexplained Recurrent Pregnancy Loss

AIMS This work aims at testing for the association of the methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms with unexplained recurrent pregnancy loss (RPL) among Egyptian women. SUBJECTS AND METHODS Participants were 70 cases having a history of two or more events of unexplained RPL and 136 controls with a good obstetric history. Detection of MTHFR C677T and A1298C mutations was done by polymerase chain reaction with restriction fragment length polymorphisms assay using restriction enzymes HinfI and MboII respectively. RESULTS Compared with controls, cases with unexplained pregnancy loss showed higher frequency of the homozygous mutant MTHFR 677 TT, 1298 CC genotypes, and the mutant haplotype 677T/1298C, although not reaching statistical significance. The frequency of 677 mutant genotypes (TT or TC) combined with either the mutant 1298 (CC or AC) or normal 1298 (AA) genotypes was significantly increased among cases with late-stage pregnancy loss versus those with early-stage pregnancy loss (p=0.001). There was also increased frequency of the 677 mutant genotypes among cases with secondary infertility compared with those with primary infertility and among cases with pregnancy loss >4 times compared with those with ≤4 times but with no statistical significance. Regarding other risk factors, it was noted that the frequency of mutations among cases with no or just one risk factor did not differ significantly from those having two or more risk factors (p=0.98). CONCLUSIONS Mutations related to the MTHFR gene are increased but not statistically significant in Egyptian women with unexplained pregnancy loss. Interaction with other genetic variants might be speculated and need to be investigated.

Association of ACE and MTHFR genetic polymorphisms with type 2 diabetes mellitus: Susceptibility and complications

Hypothesis/introduction: Polymorphisms of angiotensin converting enzyme (ACE) and methylene-tetrahydrofolate reductase (MTHFR) genes have been proposed to be associated with type 2 diabetes mellitus (T2DM) with conflicting results. This work was planned in order to check for the association of these polymorphisms with the susceptibility for and complications of T2DM among Egyptian cases. Materials and methods: This is a case controlled study involving 203 patients with T2DM and 311 healthy controls. Polymorphic variants of ACE I>D and MTHFR (677 C>T and 1298 A>C) were determined using the polymerase chain reaction (PCR) restriction analysis technique. Results: The susceptibility to T2DM was higher among subjects having the MTHFR 677TT (odds ratio (OR)=2.2, p=0.01), MTHFR 1298 AA (OR=1.84, p=0.001) and ACE (ID+II) (OR=2.0, p=0.0007) genotypes. Logistic regression analysis showed that MTHFR 677T allele was a risk factor for diabetic retinopathy (DR) (OR=3.47, p<0.001), diabetic polyneuropathy (DPN) (OR=5.2, p<0.0001) and ischemic heart disease (IHD) (OR=2.9, p<0.05), while MTHFR 1298 C allele was a risk factor for DR (OR=4.2, p<0.001) and the ACE DD genotype was a risk factor for DPN (OR=3.1, p<0.001). Conclusions: The MTHFR 677 TT genotype was associated with T2DM susceptibility and complications (DR, DPN and IHD). The MTHFR 1298 CC, AC and ACE DD genotypes were associated with DR and DPN.

Tumor necrosis factor-α gene polymorphism relationship to seminal variables in infertile men.

OBJECTIVE To assess the tumor necrosis factor (TNF)-α gene polymorphism relationship with seminal variables in fertile men (N) and those with asthenozoospermia (A), asthenoteratozoospermia (AT), and oligoasthenoteratozoospermia (OAT). MATERIALS AND METHODS A total of 50 infertile men without a female factor who were attending a fertility clinic and 48 fertile men were randomly screened for semen analysis, analysis of the TNF-α promoter region for polymorphism, seminal caspase-9, acrosin activity, α-glucosidase, and reproductive hormones. RESULTS The TNF-α GG genotype was present in 83.9%, 72.7%, 66.7%, and 59.5%, the TNF-α AA genotype in 3.2%, 6.8%, 10.4%, and 11.9%, and TNF-α AG genotype in 12.9%, 20.5%, 22.9%, and 28.6% in the N, A, AT, OAT groups, respectively. The occurrence of A allele was significantly greater among infertile patients than among fertile controls (21.6% vs 9.7%; odds ratio 0.388, 95% confidence interval 0.2 to 0.75, P = .005). Men with the TNF-α AA genotype demonstrated a significant decrease in the sperm count, sperm motility, normal sperm morphology, acrosin activity, and seminal α-glucosidase and a significant increase in seminal caspase-9 compared with those with the TNF-α GG genotype. CONCLUSION This single nucleotide polymorphism in the TNF-α(-308) gene was associated with significantly increased seminal caspase-9 and a significantly decreased sperm count, sperm motility, normal sperm morphology, acrosin activity, and seminal α-glucosidase.

Factor V Leiden and prothrombin gene mutations in Egyptian cases with unexplained recurrent pregnancy loss

Abstract Background: Thrombophilias have been suggested as a possible cause of recurrent pregnancy loss (RPL). Objective: Testing for the association of factor V Leiden (FVL) and prothrombin (FII) mutations with RPL among cases from the Nile Delta region of Egypt. Subjects and methods: Participants included 72 cases having a history of two or more events of unexplained RPL and 70 controls with a good obstetric history. Detection of FVL (G1691A) and FII (G20210A) mutations was carried out using PCR with sequence specific primers. Results: Cases showed a significantly higher frequency of FVL GA (OR = 21·38, P<0·0001) and FII GA (OR = 36·7, P<0·0001) genotypes. Cases with two or more risk factors had significant higher frequency of both mutant genotypes, while no significant difference could be elicited related to primary or secondary infertility, number of fetal losses, or phase of pregnancy loss. Conclusion: Screening for thrombophilic mutations may help in the prevention of unexplained RPL.

MTHFR C677T, A1298C and ACE I/D polymorphisms as risk factors for diabetic nephropathy among type 2 diabetic patients.

Background: Genetic variations have been proposed to play a role in the susceptibility to diabetic nephropathy. Objectives: To check for the association of genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) and angiotensin converting enzyme (ACE) genes with the development of diabetic nephropathy among type 2 diabetic patients. Methods: Participants comprised 202 patients with type 2 diabetes, of whom 102 were affected with diabetic nephropathy. Genetic variants corresponding to MTHFR C677T, A1298C and ACE I/D genotypes were determined using the PCR technique coupled with digestion and restriction analysis. Results: Cases with diabetic nephropathy had a significantly higher frequency of the MTHFR 677 TT, 677 CT, ACE DD mutant genotypes compared with diabetic cases without nephropathy. Analysis of the association of studied MTHFR C677T, A1298C and ACE I/D polymorphisms with albuminuria showed that the MTHFR 677 T polymorphism, in the recessive and dominant models, was a risk factor for both micro and macroalbuminuria, while the ACE DD mutant genotype was a risk factor for microalbuminuria and the MTHFR 1298C in the dominant model only was a risk factor for macroalbuminuria. Conclusion: These findings indicate that ACE and MTHFR genetic polymorphisms might be considered as genetic risk factors for diabetic nephropathy among patients with type 2 diabetes.

Association of cytokine gene polymorphisms with psoriasis in cases from the Nile Delta of Egypt

Background: Psoriasis is a chronic inflammatory skin disease with an immunogenetic background. This work was planned to check for the association of polymorphisms related to cytokine genes TNF-α-308 (G/A), IL-10-1082 (G/A), IL-6-174 (G/C), and IL-1Ra (VNTR) with psoriasis in cases from Egypt. Materials and Methods: This work included 46 cases with psoriasis recruited from the Dermatology Departments, University Hospitals, Nile Delta region of Egypt. They included 14 males and 32 females with an age mean ± SD of 46.68 ± 12.16 years and range of 15–70 years. Their genotypes were compared to 98 healthy controls of matched age and sex from the same locality. Genotyping was done through deoxyribonucleic acid amplification using PCR with sequence specific primers for polymorphic alleles. Results: Compared to controls, cases showed significant higher frequency of certain genotypes including IL-6-174 CC (P < 0.001, OR = 6.7), IL-10-1082 GG (P < 0.05, OR = 5.1), and TNF-α-308 GG (P < 0.05, OR = 3.7). TNF-α-308 GG and IL-10-1082 GG genotypes were higher among cases with plaque subtype of moderate severity. Combined heterozygosity for IL-10 GA, IL-6 GC with TNF GA showed a significant low frequency among studied cases. Conclusion: Genetic polymorphisms related to IL6, IL10, and TNF-α genes showed a particular pattern of association with psoriasis that may have a potential impact on disease counseling and management.

Genetic polymorphisms of NFκB1-94ins/delATTG and NFκBIA-881A/G genes in Egyptian patients with colorectal cancer

To assess the association of genetic polymorphisms of NFκB1 and NFκBIA genes with the susceptibility to colorectal cancer (CRC). Subjects included 100 Egyptian patients with CRC (60 males and 40 females) in addition to 85 healthy controls (47 males and 38 females) from the same locality. For all participants, genetic polymorphisms of NFκB1-94ins/delATTG (rs28362491) and NFκBIA-881A/G (rs3138053) were detected by using restriction fragment length polymorphism polymerase chain reaction (RFLP–PCR). CRC patients showed a significantly higher frequency of the NFκB1-94ins/ins genotype than controls (30 vs. 4.7%) that was significant in the recessive (OR 17.69, 95% CI   5.41–57.82, p < 0.0001) and codominant models (OR   18.28, 95% CI   4.87–68.6, p < 0.0001). The NFκB1-94ins allele frequency was significantly higher among patients than controls (58 vs. 39%, OR  2.18, 95% CI  1.4–3.3, p = 0.0004). We also noticed that the genotype G/G of NFκBIA-881 polymorphism was present in patients (4%) while it was absent (0%) in controls with increased frequency of the NFκBIA-881G allele in patients compared to controls (23 vs. 14%, p = 0.041). These polymorphisms were more associated with smoking and advanced tumor staging. This study indicates that the NFκB1-94ins/ins genotype was associated with the risk of developing colorectal cancer in Egyptian subjects. Also, CRC cases showed an increase in the frequency of NFκBIA-881G allele but not reaching statistical significance for multiple comparisons.