Restriction Fragment Length Polymorphism-polymerase Chain Research Articles

Interleukin-17 Genotypes in Egyptian Asthmatic Children at Zagazig University Hospitals.

Bronchial asthma (BA) is increasing among Egyptian children. It is affected by multiple factors including genetic ones. In the current study, we assessedthe relationship between interleukin-17 (IL-17) genotypes and the occurrence of BA among Egyptian children. This case-control study included 100 participants. Group I (the control group) comprised 50 healthy subjects. Group II (the asthmatic group) comprised 50 subjects diagnosed with atopic asthma according tothe Global Initiative for Asthma. Measurement of serum Ig E and eosinophilic count was performed. Detection of single nucleotide polymorphism rs2275913 of IL-17 gene by restriction fragment length polymorphism-polymerase chain reaction was conducted. GA and AA genotypes were more frequent in the asthmatic group compared tothe control group (P = 0.03 and 0.01, respectively). Subjects carrying GA and AA genotypes were more susceptibleto have asthma [odds ratio (OR) = 2.21, 95% confidence interval (CI) = 1.14-9.94, P = 0.03; OR = 7.78, 95% CI = 1.59-38.3, P = 0.01, respectively]. The A allele was higher in the asthmatic group (33%) compared tothe control group (10%). A allele carriers were more susceptibleto have asthma (OR = 4.43, 95% CI = 2.04-9.82 and P < 0.001). Immunoglobulin E (IgE) levels and eosinophil percentages were higher among the carriers of GA and AA genotypes when compared with the GG genotype. All pulmonary function tests were significantly lower among carriers of AA genotype compared with GG genotype. An A allele carrier, AA genotype, increased IgE level, and eosinophil level were significant predictors for occurrence of asthma (P = 0.01, 0.02, 0.004, and 0.01). In conclusion, AA genotype carriers and A allele carriers of the IL-17 gene are more likely to have asthma compared with controls.

The establishment of a molecular diagnostic platform for mitochondrial diseases: from conventional to next-generation sequencing

BackgroundThe aim of this study was to create a molecular diagnostic platform and establish a diagnostic pipeline for patients highly suspected of mitochondrial disorders. The effectiveness of three methods, namely, traditional restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR), Sanger sequencing for hotspot detection and whole mitochondrial DNA (mtDNA), and third-generation (Nanopore) whole mtDNA sequencing, will be compared in diagnosing patients with suspected primary mitochondrial diseases (PMDs). The strengths and limitations of different methods are also discussed. Material and methodsA single-center prospective cohort study was conducted to validate the diagnostic pipeline for suspected mitochondrial diseases. In the first stage, a PCR-based method with five sets of primers was used to screen for eight hotspots (m.3243A>G, m.3460G>A, m.8344A>G, m.8993T>G, m.9185T>C, m.11778G>A, m.13513G>A, and m.4977deletion) using either RFLP or direct Sanger sequencing. Sanger sequencing was also used to confirm the RFLP-positive samples. In the second stage, for samples with negative screening results for the eight hotspots, mitochondrial whole-genome sequencing was performed using Sanger sequencing or third-generation nanopore sequencing. ResultsBetween June 2020 and May 2023, 30 patients from ages 0 to 63 with clinically suspected mitochondrial disease were enrolled. The positive yield for the diagnosis of PMDs was 8/30=26.7%, and the sensitivity of the heteroplasmy level for the RFLP-based method was approximately 5%. The remaining 22 patients who tested negative at the first stage were tested using Sanger sequencing or the third-generation sequencing Nanopore, and all tested negative for pathological mtDNA mutations. Compared to the Sanger sequencing method, the results of RFLP-PCR were compromised by the limitations of incomplete RFLP enzyme digestion. For whole-genome sequencing of mtDNA, Sanger sequencing, instead of nanopore sequencing, is preferred at our institution because of its cost-effectiveness. Conclusions: In our highly selective cohort, most tested positive in the first stage of the 8 hot spots screen. Sanger sequencing is a conventional and accurate method for mitochondrial disease screening, at least for the most common hot spots in the region. The results revealed that Sanger sequencing is an accurate method with the benefit of being more cost-effective. This integral platform of molecular diagnosis bears the advantages of being relatively low cost and having a shorter reporting time, facilitating crucial identification of patients with clinical evidence of such disorders. This diagnostic flowchart has also been translated into routine clinical use in the tertiary hospital.

Association of polymorphism of IL-17A, IL-17F, and IL-6 with Toxoplasma gondii infection susceptibility in HIV/AIDS patients in Shiraz, southern Iran.

Toxoplasma gondii infection is considered as one of the most important opportunistic infections and cause of death in HIV patients. In this cross-sectional study, 334HIVpositive patients were included. The molecular test was performed by the restriction fragment length polymorphism-polymerase chain reaction method. Allelic frequency, haplotype analyses, and linkage disequilibrium were calculated. The odds ratio was calculated. The linear regression model was used to analysis of interleukin (IL)-17A, IL-17F, and IL-6 single-nucleotide polymorphism genotypes in HIV patients with and without toxoplasmosis. In total, 95 tested'patients (28.4%) were positive for toxoplasmosis. The risk of toxoplasma infection in the current study did not correlate with IL-17 and IL-6 polymorphism and the risk of contracting toxoplasma was also not significantly correlated in this study. There was no association between the frequency of alleles and the risk of toxoplasma infection in IL-17 haplotype analysis. The findings of this study revealed that there were significant differences in the serum levels of IL-6 and IL-17A, but not IL-17F, between the case and control groups in various genetic models. However, these polymorphisms did not show a significant relationship with toxoplasma infection in HIV-positive patients. This study represents the first investigation in Iran to explore the role of IL-6 and IL-17 polymorphisms in toxoplasma infection among HIV-positive patients.

Probiotics Potential of Lactic Acid Bacteria from Horse Milk in Tambora, West Nusa Tenggara, Indonesia

This study explores the probiotic potential of lactic acid bacteria (LAB) in horse milk from Tambora, West Nusa Tenggara by examining its functional and probiotic attributes, including bile salt tolerance, low pH, simulated gastric juice (SGJ), simulated intestinal juice (SIJ), antibacterial activity, as well as bile salt hydrolase (BSH) genes. Genotyping of LAB was performed using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) analysis and then identified based on the 16S rRNA gene. A total of 25 LAB isolates showed the ability to grow at low pH, tolerant to bile, and survived under SIJ and SGJ conditions. The BSH gene was confirmed in three isolates, namely: SK1-28, SK2-30, and SK2-34. Results of RFLP-PCR analysis showed that the LAB isolates were grouped into three groups based on the number and molecular weight of the differences DNA fragments. The 16S rRNA analysis showed that the first two groups were Lacticaseibacillus rhamnosus, whereas the third group was Lactiplantibacillus plantarum. In addition, all Lacticaseibacillus rhamnosusisolates in group I showed the ability to grow at pH 9.0, but not group II. It can be concluded that Lacticaseibacillus rhamnosus and Lactiplantibacillus plantarum can be used as the indigenous probiotic bacteria source from Indonesia.

Relationship between interleukin 17 polymorphism in rs 2275913 and rs 763780 and interleukin 6 in rs 1800795 gene region with sensitivity to antiretroviral drugs in patients with human immunodeficiency virus

Drug complication is still considered as one of the most important causes of death and drug in-compliance around the world. In this cross sectional study, 372 people living with HIV (PLHIV) above 16 years were enrolled. The drug complication was extracted based on the information of the patient's file. The molecular test was performed by the Restriction Fragment length polymorphism-Polymerase Chain Reaction method. Allelic frequency, haplotype analyses, linkage disequilibrium and odds ratio (OR) were calculated. The linear regression model was used to analyze the association of IL’SNPs with drug complication after adjustment for age and sex. Drug complications were observed in 150(40.3%) participants. The most common drug complications were hematological 94(62.7%) ones. The SNPs- rs 2275913 and rs763780- of IL-17were in complete linkage (D́ = 1 and r = 1). A-A haplotype of IL-17 in SNPs- rs 2275913 and rs763780 can increase the risk of drug complication up to 1.628 times more than other haplotypes and G-G and G-A haplotypes have a protective role among them 0.268 and 0.628 times, respectively. Our result for the first time demonstrated the role of IL-17 polymorphism in induced antiretroviral drug complication incidence. Probably A-A haplotype could increase the immune response to anti-retroviral drugs, and G-G and A-G haplotypes can decrease it.

Association between myeloperoxidase polymorphism (MPO-129G/A), oxidized low-density lipoprotein level, and atherogenic indices in patients with atherosclerosis; a case-control study

ObjectiveThis study aimed to evaluate the correlation between myeloperoxidase polymorphism (MPO-129G/A) and the risk of atherosclerosis and its association with oxidized low-density lipoprotein (Ox-LDL) levels and atherogenic indices. Materials and methodsThe present study was performed on 255 subjects, including 125 controls and 130 atherosclerotic patients. To determine MPO-129G/A polymorphism, the restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) method was utilized. Serum lipid profile levels and fasting blood sugar (FBS) level were calculated using an auto-analyzer. Atherogenic indices and Ox-LDL levels were measured using formulae and ELISA method, respectively. ResultsThe findings illustrated no considerable difference between the patient and control groups concerning MPO-129G/A genotypes (p > 0.05). There was no substantial relationship between MPO-129G/A genotypes, Ox-LDL levels, and atherogenic indices in the patient group (p > 0.05), while patients possessed considerably higher Ox-LDL level and atherogenic indices (p < 0.05). In addition, CRI-II (LDL/HDL) was significantly correlated with Ox-LDL (p = 0.001) and FBS (p = 0.0007). ConclusionMPO-129G/A indicated no association with Ox-LDL levels and atherogenic indices. However, atherosclerotic patients had higher Ox-LDL levels and atherogenic indices. Furthermore, CRI-II was significantly correlated with levels of Ox-LDL and blood glucose, suggesting a strong marker for predicting the risk of atherosclerosis.

The Association between Apolipoprotein E Genotypes and Serum Malondialdehyde Level with End-Stage Renal Disease.

The Apolipoprotein E (ApoE) polymorphism plays an important role in the pathophysiology of end-stage renal disease (ESRD). Additionally, ApoE may contribute to the progression of oxidative stress. Thus, this study aimed to determine the ApoE gene polymorphism and evaluate the malondialdehyde (MDA) level in ESRD patients and healthy individuals. The present cross-sectional study was conducted at 2010 at Kermanshah University of Medical Sciences (Kermanshah, Iran). The study population comprised ESRD patients (n=136) and healthy individuals (n=137). The MDA level was assessed using high-performance liquid chromatography (HPLC), and the frequencies of ApoE gene alleles were analyzed using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). The data were analyzed using Statistical Package for Social Sciences (SPSS), version 13. The significant differences of ApoE genotypes in case and control groups were assessed using Pearson's Chi square tests, and two-tailed Student's tests. A logistic regression model was used to calculate the odd ratio. P<0.05 was considered statistically significant. According to the results, ESRD patients had a higher frequency of the E2/E3 genotype than the healthy group (P<0.001). The results indicated that E3/E4 genotype frequency in the patients' group was higher than that of the control group (P=0.026). Furthermore, the E3/E2 (OR=5.7, 95% CI=2.68-12.14) (P<0.001) and E3/E4 (OR=1.57, 95% CI=1.05-2.34) (P=0.029) genotypes were found to increase the risk of ESRD. Moreover, the MDA level in ESRD patients was higher than the healthy individuals (P<0.001). The patients with E3/E2 (P<0.001) and E3/E4 (P<0.001) genotypes had a higher level of MDA than the control group. According to the findings, patients with ESRD had higher genotypes of E3/E2 and E3/E4, which suggests a higher risk of developing ESRD.

Evaluation of Vitamin D Receptor Taq I (rs731236) and Bsm I (rs1544410) Gene Polymorphisms in Patients with Chronic Kidney Disease-mineral and Bone Disorder

BACKGROUND: Chronic kidney disease-mineral and bone disorder (CKD-MBD) is one of the main culprits of increasing morbidity and mortality in patients with stage: 3–5 CKD. Association between Vitamin D receptor (VDR) genetic polymorphisms and CKD-MBD has been inconsistent. AIM: The aim of the study was to assess the association of VDR Taq I (rs731236) and Bsm I (rs1544410) gene polymorphisms with CKD; and with the development and progression of CKD-MBD. METHODS: Sixty adult (43–56 years.) Egyptian CKD-MBD male patients (CKD stages: 3–5 with estimated glomerular filtration rate &lt;60 mL/min 1.73 m2) and 30 matched-pair healthy controls were recruited from Theodor-Bilharz Research Institute. Bsm I and Taq I polymorphisms of VDR gene were assessed using restriction fragment length polymorphism-polymerase chain reaction. RESULTS: CKD-MBD patients having Taq I “tt” mutant gene had a significant decrease in serum 25 hydroxy Vitamin D and a significant elevation of plasma intact parathyroid hormone levels. Having the homotypic “tt” gene variant of VDR Taq I increased the susceptibility to CKD-MBD (Odds ratio [OR]: 19.6, CI: 4.3–89.9 p &lt; 0.01) compared to having the wild “TT” or heterotype “Tt” genotype. Moreover, presence of VDR Taq I “tt” genotype increases OR of having 25 hydroxy Vitamin D deficiency in CKD-MBD patients 7.25 times (CI = 2.21–23.80; p &lt; 0.01). CONCLUSION: VDR Taq I (rs731236) “tt” genotype increases the susceptibility to CKD-MBD development and progression in Egyptian CKD patients. Moreover, the presence of Taq I “tt” genotype in CKD-MBD patients is independently associated with the risk of developing Vitamin D deficiency.

Effects of ABCG2 C421A and ABCG2 G34A genetic polymorphisms on clinical outcome and response to imatinib mesylate, in Iranian chronic myeloid leukemia patients

BackgroundChronic myeloid leukemia (CML) is a multifactorial clonal myeloid neoplasm that mainly arises from the Philadelphia chromosome. Even though imatinib mesylate (IM) is considered the gold standard for first-line treatment, a number of CML patients have shown IM resistance that can be influenced by many factors, including pharmacogenetic variability. The present study examined whether two common single nucleotide polymorphisms (SNPs) of ABCG2 (G34A and C421A) contribute to IM resistance and/or good responses.Material and methodsA total of 72 CML patients were genotyped with high-resolution melting (HRM) and restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). We also determined the cytogenetic and hematological response, as evaluable factors for measuring response to imatinib.ResultsIn the current study, we explored the relationship between the different variants of ABCG2 G34A and C421A and clinical response to imatinib among CML patients. There were no statistically significant differences between genotypes of C421A and G34A and allele frequencies among the resistant and responder groups, with response to IM (P > 0.05). Also, we found no statistically significant association between genotypes and cytogenetic and hematological responses.ConclusionThis is the first study to investigate the association between genotypes of the G34A and C421A SNPs and the outcome of IM treatment in Iranian population. As a whole, genotyping of these SNPs is unhelpful in predicting IM response in CML patients.

Serum human β-defensin-1 (hBD-1) and -20G/A DEFB1 gene polymorphism in Behçet’s disease patients: Relation to clinical characteristics

Aim of the work:Behçet’s disease (BD) is a multisystemic inflammatory disease of unknown cause. Beta defensins (β-defensins) are endogenous antimicrobial peptides that have the ability to kill or inactivate a wide spectrum of bacteria and fungi by changing their membranes stability. It also plays a significant role in inflammation owing to its chemotactic activity for T cells and immature dendritic cells. To explore whether β-defensins might be involved in BD pathogenesis, serum human β-defensin-1 (hBD-1) and defensin-β-1 (DEFB1)-20 G/A gene polymorphism were assessed in patients with BD. Patients and methodsThe study included 40 BD patients and 40 matched controls. Serum hBD-1 was assessed by enzyme-linked immunosorbent assay (ELISA) and the -20G/A genotypes (rs11362) were determined by restriction fragment length polymorphism-polymerase chain reaction(RFLP-PCR) analysis. ResultsThe mean age of patients was 33.5 ± 10.8 years. They were 32 males and 8 females (M:F 4:1). Disease duration was 8.5 ± 7.3 years. In patients, the median serum hBD-1 concentration was 295.5 pg/ml (27.6–1776.6 pg/ml) and was significantly higher than in the control (median 129.6 pg/ml; 23.7–986.3 pg/ml) (p < 0.001). Serum hBD-1 significantly correlated with C-reactive protein (r = 0.85, p = 0.001) and platelets (r = 0.4, p = 0.012). No significant difference was found between patients and control regarding genotyping of -20G/A DEFB1 (rs11362). Skin manifestations were significantly higher among patients with GA genotype 18/23 (78.2 %) than patients with AA genotype 7/16 (43.8 %)(p = 0.027). However, there was no association between serum hBD-1 and −20A/G polymorphism. ConclusionsThe hBD-1 is implicated in the pathogenesis of BD and may be a possible therapeutic target.

Explore the distribution of (rs35742686, rs3892097 and rs1065852) genetic polymorphisms of cytochrome P4502D6 gene in the Moroccan population

BackgroundThe CYP2D6 gene encodes a crucial enzyme involved in the metabolic pathways of many commonly used drugs. It is a highly polymorphic gene inducing an interethnic and interindividual variability in disease susceptibility and treatment response. The aim of this study is to evaluate the frequency of the three CYP2D6 most investigated alleles (CYP2D6*3, CYP2D6*4, and CYP2D6*10 alleles) in Morocco compared to other populations.This study enrolled 321 healthy Moroccan subjects. CYP2D6 genotypes and allele frequencies were assessed using a restriction fragment length polymorphism–polymerase chain reaction genotyping method. The Principal Component Analysis (PCA) and dendrogram were conducted to evaluate genetic proximity between Moroccans and other populations depending on CYP2D6 allele frequencies.ResultsAccording to the current study, the results observed the homozygous wild type of the three studied SNPs were predominant among the Moroccan population, while 1.4% of Moroccans carried the CYP2D6*4 allele responsible for a Poor Metabolizer phenotype and associated with low enzyme activity which may induce a treatment failure. The PCA and cluster dendrogram tools revealed genetic proximity between Moroccans and Mediterranean, European and African populations, versus a distancing from Asian populations.ConclusionThe distribution of CYP2D6 polymorphisms within Morocco follows the patterns generally found among the Mediterranean, European and African populations. Furthermore, these results will help to lay a basis for clinical studies, aimed to introduce and optimize a personalized therapy in the Moroccan population.

Protective Role of IL7A-197 A/G Heterozygosity in the Development and Severity of Colorectal Cancer in the Bulgarian Population.

Background: Interleukin (IL)-17A and IL-17F, expressed mainly by a novel subset of CD-positive (+) T-helper (Th) cells of the immune system, has been closely related to inflammatory conditions underlying colorectal cancer pathogenesis. Accordingly, we conducted a case-control study to investigate the association of common single nucleotide polymorphisms (SNP) in the IL17A and IL17F genes (rs2275913 and rs763780, respectively) with the susceptibility and severity of CRC patients from the Bulgarian population. Methods and Materials: 136 patients with histologically confirmed CRC diagnosis and 116 healthy individuals were recruited in the present study. Genotypes were determined by the restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique. Results: The IL17A heterozygous A/G-genotype was overrepresented among the control group (p = 0.003). Additionally, the carriers of the heterozygous A/G-genotype had a 2.39-fold lower risk for CRC compared to the G/G-genotype (OR = 0.418, p = 0.006). Our results also indicated that in the advanced CRC stages (III + IV) the heterozygous genotype (A/G) appeared to be less frequent (p = 0.024, χ2-test). Among the patients with detected distant metastases, the A/G-carriers were the smallest part (14.3%) compared to the homozygous genotypes A/A (42.9%) and G/G (42.8%), p = 0.006. There was no association of the studied IL17F rs763780 SNP with susceptibility and severity of CRC among the studied subjects, although the heterozygous C/T-carriers had shorter median survival compared to the T/T-carriers (p = 0.129). Conclusions: Our study finds a protective role of heterozygosity for the IL17A-197A/G SNP and negative effects of the A-allele on CRC progression.

Vitamin D receptor polymorphism and prostate cancer prognosis.

Prostatic epithelial cells synthesize the active form of vitamin D (1,25-dihydroxyvitamin D3), which participates in regulating prostate growth. Calcitriol, a synthetic form of vitamin D3, exhibits antiproliferative and prodifferentiation activities in prostate cancer. The function of 1,25-dihydroxyvitamin D3 is mediated by its binding to vitamin D receptor (VDR). VDR forms a heterodimer, typically with retinoid X receptor, to regulate vitamin D target genes. We evaluated the relationship between VDR polymorphism and clinical characteristics associated with prostate cancer risk and prognosis among Egyptian men. This case-control study included 2 groups of patients: group A, a control group of 50 subjects with benign prostate hyperplasia, and group B, 50 subjects newly diagnosed with prostate cancer. All participants performed complete blood count, liver and kidney function tests, prostate specific antigen measurement, histopathological analysis and immunohistochemistry for Dickkopf Homolog 3. Restriction fragment length polymorphism-polymerase chain reaction as performed to detect VDR polymorphism. Patients with prostate cancer and controls showed a significantly different CA genotype frequency (p = 0.007). Furthermore, prostate-specific antigen levels were significantly different in different genotypes in patients with prostate cancer (p < 0.001). Finally, T stage and the VDR ApaI C/A polymorphism were significantly associated (p < 0.041). The VDR ApaI C/A polymorphism may be a diagnostic and prognostic marker for prostate cancer in Egyptian men.

Genetic Polymorphism in Toll-Like Receptor 3 and Interferon Regulatory Factor 3 in Hepatitis C Virus-Infected Patients: Correlation with Liver Cirrhosis.

Host genetic factors could play a primary role in determining the risk for cirrhosis development in chronic hepatitis C virus (HCV)-infected patients. We designed this work to study the effect of single-nucleotide polymorphism (SNP) in Toll-like receptor 3 (TLR3) and interferon regulatory factor (IRF) on the risk of HCV-related cirrhosis. This study enrolled 139 Egyptian HCV-infected patients. They were divided into patients with cirrhotic (56) and noncirrhotic (83) liver. Genotyping of rs3775291 F459F (+1234C/T) and rs3775290 L412F (+1377C/T) in TLR3 and IRF3 rs2304204 (-925A/G) was performed by restriction fragment length polymorphism-polymerase chain reaction. Although there is no significant difference in genotype and allele distribution of +1377C/T of TLR3 gene between cirrhotic and noncirrhotic subjects, CC (odds ratio [OR] = 1.572, 95% confidence interval [CI]: 0.781-3.164); TT (OR = 1.463, 95% CI: 0.351-6.104) genotypes might be considered as risk factors for liver cirrhosis. On the contrary, the analysis revealed that only one genotype (CC) and one allele (C) were detected in +1234C/T SNP, with the total disappearance of CT/TT genotypes and T allele in all subjects. On the contrary, lower frequency has been found for the AG genotype of the IRF3 (-925A/G) gene in cirrhotic patients compared with noncirrhotic ones, indicating that AG is a protective genotype (OR = 0.509, 95% CI: 0.256-1.012). Our data stressed the association of AG genotype SNP in IRF3 (-925A/G) in protection against the worth outcome of HCV infection.

Association between ATP binding cassette gene member 1 polymorphism and glucocorticoid response in children with immune thrombocytopenia.

ATP binding Cassette gene member 1 (ABCB1) polymorphism has been incriminated in susceptibility to many malignant, infectious and autoimmune diseases. Recently, it was reported that ABCB1 polymorphisms might have a link to disease progression as well as response to therapy. We aimed to study the association between ABCB1 gene polymorphism and glucocorticoid response in children with newly diagnosed immune thrombocytopenia (ITP). A case control study was conducted on 90 newly diagnosed children with ITP and 90 healthy controls over a period of 1 year. ABCB1 (C3435T) polymorphism was determined by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) in patients and controls. There was no significant difference between patients and controls as regards to frequency of different ABCB1 genotypes (CC, CT, and TT genotypes were 44.4%, 36.7%, and 18.9% respectively in patients and 48.9%, 38.9%, and 12.2% respectively in controls, P value = 0.18). 80% of patients who received steroids alone or steroids in combination with intravenous immunoglobulin showed complete recovery. There was highly significant relationship between ABCB1 genotypes and response to steroids where 55 % of responders had CC (wild) genotype while 40 % of nonresponders had TT (mutant) genotype. We concluded that ABCB1 gene polymorphism may contribute to the response to steroids in Egyptian children with ITP where patients with homozygous CC genotype responded better to steroids than patients with homozygous TT genotype. These results may help us choose the appropriate initial treatment in these children.

Role of matrix metalloproteinase 9 gene polymorphism as genetic risk factor for lung cancer

Lung cancer (LC) is one of the most incident malignancies worldwide. This study investigated the potential role of matrix metalloproteinase9 gene (MMP9) single nucleotide polymorphisms (SNPs) in Lung cancer patients. The study population was included 75 Iraqi patients with confirmed Lung cancer and 25 healthy people as controls. In this study, the -1562 C/T polymorphism in MMP9 gene was detected by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique. The genotype distribution results of the -1562 C/T SNP of MMP9 gene have shown no significant differences (p&gt;0.05) between controls (CC: n= 15, 60%; CT: n=9, 36%; TT: n=1, 4%) and Lung cancer patients (CC: n= 44, 58.66%; CT: n=30, 40%; TT: n=1, 1.34%). Results of the association analysis between genotype distribution of the -1562 C/T SNP of MMP9 gene and clinicopathological criteria of cases such as histological grading, TNM staging and lymph node metastasis, had shown no significant association (p&gt;0.05). While analysis of the association between genotype distribution of the -1562 C/T SNP of MMP9 gene and smoking status among patients were statically significant (p&lt;0.05). These results suggested that the SNP in MMP9 gene was involved in lung cancer progression, tissue invasion and metastasis involvement.

Association of the genetic variants of XRCC1 DNA repair gene as genetic factors for breast cancer in Iraqi patients

Breast cancer (BC) is the second leading cause of cancer deaths worldwide and the most common type of cancer among women. This study was included blood of 75 Iraqi women with confirmed breast cancer and 25 samples with normal breast tissues were considered as control group. The restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique was performed the detection and genotyping for polymorphisms of X-Ray Repair Cross Complementing 1 (XRCC 1) gene Arg194Trp. The genotype distribution of the XRCC 1 Arg194Trp showed significant difference (p&lt;0.05) between controls (Arg/Arg: n= 25, 100%; Arg/Trp and Trp/Trp n=0) and breast cancer patients (Arg/Arg: n= 47, 62.67%; Arg/Trp: n=2, 2.67%; Trp /Trp: n=26, 34.67%). The result showed an increased in Trp/Trp genotype and Trp allele of the XRCC1 Arg194Trp in breast cancer patients than control.

Association of matrix metalloproteinase-8 gene promoter polymorphisms with type II diabetes mellitus in Syrian population

ObjectivesMatrix metalloproteinase (MMP)-8 is an enzyme that cleaves extracellular matrix (ECM). It is involved in many diseases' pathology including diabetes mellitus type II (DM). In MMP-8 gene promoter region, there are many polymorphisms with an important role in gene transcription modifying and protein production. -381A/G and -799 C/T SNPs are the most effective polymorphisms, and our study is the first work aimed to evaluate the association between these polymorphisms and DM susceptibility in Syrian population. Designstudy sample included 140 subjects allocated into two groups: H= healthy control, DM= diabetes mellitus. MMP-8 SNPs were determined by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique. ResultsThe frequencies of genotypes were significantly different between study groups (adjusted OR =3.621, 95% CI: 1.391-9.428, P = 0.008 for -381GG genotype), and (OR =4.941, 95% CI: 1.640-14.890, P = 0.005 for -799TT genotype). Subjects with minor alleles seem to be more likely to develop DM more than those with the major alleles (OR=2.842, P = 0.01 for G) and (OR =3.995, P = 0.001 for T). ConclusionThis study data has illustrated that there could be a remarkable association between MMP-8 gene promoter polymorphisms -381A/G and -799C/T and diabetic susceptibility in Syrian population.

IL-17A rs2275913 gene polymorphism in patients with diabetic nephropathy

Introduction: Diabetic nephropathy (DN) is a major microvascular complication of diabetes, leading to end-stage renal disease (ESRD). Objectives: The aim of this study was to investigate the association between the rs2275913 polymorphism of interleukin-17 (IL-17) gene and the incidence of kidney disease in patients with type 2 diabetes (T2D). Patients and Methods: Blood samples were collected from 113 T2D patients including 56 patients with nephropathy and 57 patients without nephropathy. In addition, 150 healthy individuals were included in this study to compare the results. Gene study was conducted by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) method. Results: The frequency of A allele of IL-17A (rs2275913) gene polymorphism was significantly higher in patients with DN compared to healthy controls (P=0.043). In addition, serum creatinine levels were significantly higher in DN patients regardless of their genotypes (P&lt;0.001). Conclusion: Our results showed that diabetic patients who carry at least one A allele of IL-17A (rs2275913) gene polymorphism may develop DN.

Interferon-Gamma (IFNg) +874A/T Polymorphism Does Not Significantly Affect the Severity of Periodontitis.

Objectives Interferon-gamma (IFNg) is an immune-regulatory cytokine with a role in host responses to periodontitis. Genetic factors have been reported to modify the corresponding protein expression. The objective of this study was to evaluate the association and role of IFNg polymorphisms, such as IFNg +874 A/T, and the susceptibility to periodontitis. Materials and Methods A total of 100 unrelated subjects were included in the present study. Genomic deoxyribonucleic acid (DNA) was obtained from peripheral blood of 43 patients with mild periodontitis and 57 patients with severe periodontitis. The determined clinical parameters of periodontitis included probing depth, clinical attachment loss, and papilla bleeding index. The oral hygiene indicators were also assessed. The level of IFNg was determined from the gingival crevicular fluid by enzyme-linked immunosorbent assay technique. The IFNg +874 A/T polymorphisms were analyzed from peripheral blood by the method of restriction fragment length polymorphism-polymerase chain reaction. Statistical Analysis Statistical analysis of the results was conducted using chi-squared testing for categorical data. Independent t -tests and Mann–Whitney U tests were used for numeric data. Kruskal–Wallis testing was used to compare genotypes concerning for IFNg +874 A/T polymorphism. A p -value < 0.05 was assumed for statistical significance. Results Analysis of the IFNg +874 A/T polymorphism showed no significant differences with the level of IFNg. No significant differences were observed either in IFNg +874 A/T polymorphism between the subjects with mild periodontitis and those with severe periodontitis ( p > 0.05). The subjects with severe periodontitis showed marginally but not significantly higher levels of IFNg compared with subjects with mild periodontitis ( p > 0.05). Conclusion The polymorphism of IFNg +874 A/T was not associated with the level of IFNg nor with the risk of periodontitis in this study.