PCR-RFLP Technique Research Articles

Association of Methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism in breast cancer patients of Bihar: Case-control study

Breast cancer (BC) is the most frequent cancer in women globally and the main cause of cancer-related mortality. MTHFR C677T is a functional polymorphism in the genes encoding a one-carbon metabolism enzyme that affect folate metabolism and may be associated with breast cancer susceptibility. We have conducted a case-control study taking 100 cases of diagnosed breast cancer and 100 healthy control. PCR-RFLP technique was used to detect the genotype of C677T polymorphism in the study population.About 35% of cases had normal genotype(C677C), 49% had the genotype C677T & rest 16% had the genotype T677T for MTHFR. Among control the frequency of C677C genotype was 55% C677T was 40% & T677T was 5%. C677T & T677T MTHFR genotype were more common in case as compared to control [χ2 (2, N = 200) = 11.12, p = 0.004]. T allele frequency was significantly higher in case (40%) as compared to control (5%) [χ2 (1, N = 400) = 10.91, p = 0.001]. Cases with MTHFR 677TT genotype had a significantly higher risk of BC than 677CC individual [OR=1.64 (1.03 to 2.10)] and increasing T-allele was significant in BC [OR=1.07 (0.71 to 1.33)] (P for trend = 0.04).C677T of MTHFR gene has a strong correlation with breast cancer in the studied population and can be used as a prognostic marker for the development of breast cancer.

Microscopic and Molecular Detection of Sarcocystis cruzi (Apicomplexa: Sarcocystidae) in the Heart Muscle of Cattle.

Sarcocystis is a genus of coccidian protozoa that at least seven species of it can parasitize cattle. The global prevalence of Sarcocystis is close to 100% in adult cattle. The main aim of this study was to identify the infection rate of Sarcocystis spp. in heart of cattle in Tehran, Iran by microscopy and PCR-RFLP methods. Totally, 100 bovine heart samples were collected from the main slaughterhouse of Shahriar, Meysam slaughterhouse, west of Tehran in 2016. At first, heart samples were completely examined for the presence of sarcocystic macrocysts. Then, for microscopic examination, 50 g of each heart was digested in sterile condition using pepsin acid digestion method. Then, the species of the parasite were detected by PCR-RFLP technique and sequencing. Overall, 97 of 100 of the heart muscle samples were infected with Sarcocystis. All the samples were detected as S. cruzi through similar patterns in PCR-RFLP. S. cruzi is the most common species in the heart of cattle slaughtered in Shahriar.

Molecular analysis of acid-stable alpha-amylase (asAA) gene in Aspergillus niger using PCR-RFLP

Introduction and Aim: Amylase is an important enzyme with vast applications in various industries such as food and therapeutic industries. Aspergillus niger is commercially engaged in the making of alpha-amylase. Acid-stable alpha -amylase is mostly produced with microorganisms such as Bacillus and Aspergillus. The aim of this research was the molecular investigation of the acid-stable alpha-amylase (alpha-sAA) gene in A. niger. Materials and Methods: Sixty-three A. niger isolates were evaluated in this study. PCR method was performed for amplification of a 347 bp DNA band of the alpha-sAA gene. The Hpa II Restriction endonuclease was used for the digestion of PCR fragments. Results: A 347 bp DNA fragment was recovered from 49 out of 63 (78%) isolates. After cutting the PCR products with the HpalphaII enzyme, 81.6% of isolates showed the expected band and 18.4% presented different restriction endonuclease patterns. Conclusion: The results demonstrated the PCR-RFLP technique performed in this research was a valuable tool for analysis of the alpha-sAA gene in A. niger isolates.

Identification of protein profile in metacyclic and amastigote-like stages of Leishmania tropica: a proteomic approach

Leishmaniasis is a tropical disease that leads to various clinical phenotypes. This study aimed to investigate protein expression changes in metacyclic and amastigote-like stages of L. tropica isolated from Iranian cutaneous leishmaniasis patients. Isolated samples were cultured and species type identified using PCR–RFLP technique. The promastigotes were grown in RPMI1640 media and differentiated to metacyclic and amastigote-like forms, followed by the extracted proteins of both successive stages carried out for proteomics and bioinformatics analysis. Using SWATH-MS quantitative proteomics technique, a total 176 and 155 distinct proteins were identified in metacyclic and axenic amastigote stages, respectively. Of these, 65 proteins were altered significantly (p-value < 0.05 and fold change ≥ 2) between studied stages. Several gene ontology (GO) categories were enriched for biological process during conversion of metacyclic promastigotes into amastigote-like, which “metabolic process” (GO: 0044281, P-Value: 6.52e-5), and “translation” (GO: 0006412, p-value: 5.01e–14) were disclosed as the top category in up and down-regulated proteins, respectively. Also, the KEGG pathway analysis indicated “metabolic pathways” and “ribosome” term as the most important pathways in up and down-regulated proteins, respectively. According to protein interaction network analysis, enolase (ENOL) has been detected as main hub proteins during differentiation, followed by Putative NADH-dependent fumarate reductase (LmjF.35.1180) and 40S ribosomal protein S2 (LmjF.32.0450). Overall, protein changes possibly play important roles in L. tropica biology. Anabolic pathways were down-regulated, whereas catabolic pathways were up-regulated during L. tropica differentiation. These protein expression changes could provide parasite survival in host macrophages, and could use as novel potential drug and vaccine targets for leishmaniasis.Graphical

Association Of the Methylene Tetrahydrofolate Reductase (MTHFR) Gene Polymorphism with Susceptibility to Recurrence of Cardiovascular Outcomes Among Ischemic Stroke- A Randomized Controlled Trial

Background: Hyperhomocystenemia and genetic variants are factors for causing young age stroke globally. This study aims to identify homocysteine related-MTHFR gene polymorphism that associated with recur-rent cardiovascular outcomes. Methodology: A randomized controlled trial conducted upon 90 hyperhomocysteinemic ischemic stroke patients were taken from the neurology wards of a tertiary care hospital were randomly selected into vita-min B therapy group and control groups (n=45 in each group). Baseline subject details were collected ve-nous blood sample for MTHFR genetic testing via PCR-RFLP technique along with blood homocysteine lev-els, vitamin B12, folic acid levels. Results: The results showed that the frequency of CT genotype polymorphism was 15.5% vs 13.3% for the MTHFR C677T gene without any significant difference between vitamin group and control group respective-ly (p-value &gt;0.05). The reduction in mean homocysteine up to -6.77±4.50 versus -2.08±0.71 µmol/L in the vitamin group as compared to control group respectively, p value 0.001. Conclusion: Considerable amount of MTHFR gene polymorphism found among hyperhomocysteinemic is-chemic stroke of sub-Himalayan region. Nutritional deficiencies including vitamin B 12 &amp; folic acid, and some hidden reasons found, which could lead to the primary cause of hyperhomocysteinemia. Vitamin B therapy is an effective for reducing homocysteine.

Development of a simple PCR-RFLP technique for detection and differentiation of E. histolytica, E. dispar and E. moshkovskii.

Epidemiological studies on amoebic infections are complicated by morphological overlap between the pathogenic E. histolytica, the commensal E. dispar and the amphizoic E. moshkovskii, necessitating molecular identification. The present study developed a simple and economical 18S PCR-RFLP method for the simultaneous detection and differentiation of the three species. PCR products were differentiated by Tat1 restriction digestion generating three different RFLP patterns. Validation was conducted by screening 382 faecal samples from human patients from Kolkata, India, hospitalized for diarrhoea. Analysis indicated that the PCR-RFLP could successfully differentiate between the three species and was confirmed by sequence analysis. This method could prove useful for clinical and epidemiological studies of amoebiasis.

The association between PD-1 gene polymorphisms and susceptibility to multiple sclerosis

Programmed cell death 1 (PD-1) is an immune checkpoint and has been reported to be associated with several autoimmune diseases. We aimed to investigate the association between human PD-1 gene (PDCD1) polymorphisms and multiple sclerosis (MS). This case-control study was conducted on 229 MS patients and 246 healthy controls. Genotyping of rs36084323 (PD-1.1 G/A), rs11568821 (PD-1.3 G/A) and rs2227981 (PD-1.5 C/T) polymorphisms was performed by PCR-RFLP technique. The frequency difference of PD-1.1 genotypes and alleles (−536 G/A) between patients and healthy controls was not significant. Regarding PD-1.3, the AA + AG genotype was found to be relatively higher in the control group. Concerning PD-1.5 (+7785 C/T), the frequency of T allele carriers (TT + CT) was relatively higher in MS patients, which was marginally insignificant (p = .07). PD-1 gene polymorphisms may be associated with MS; however, accurate conclusions require further studies with a larger number of samples.

Association investigations between ACE1 and ACE2 polymorphisms and severity of COVID-19 disease.

Due to the unique affinity of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to the angiotensin-converting enzyme 2 (ACE2) receptor in patients, the foremost recent evidence indicated that ACE1 and ACE2 polymorphisms could affect the susceptibility of individuals to SARS-CoV-2 infection and also the disease outcome. Here, we aimed to assess the possible association between two polymorphisms and the severity of disease in patients. In the present study, 146 patients with COVID-19 who were admitted to the Mazandaran University of Medical Sciences hospitals between March 2020 and July 2020 were enrolled in this case–control study. The patients were divided into four groups based on clinical symptoms and severity of the diseases (mild, moderate, severe, and critical). After DNA extraction, the ACE gene I/D polymorphism (rs4646994) and ACE2 gene polymorphism (rs2285666) were genotyped using Gap-PCR and PCR–RFLP techniques, respectively. Then, five samples from each obtained genotype were confirmed by Sanger sequencing technique. Data were analyzed with SAS software version 9.1 using appropriate statistical procedures. The ACE gene I/D polymorphism (rs4646994) genotypes were classified into three types: I/I, I/D, and D/D. Our finding indicated that the prevalence of ACE1 D/D genotype was significantly higher in severe and critical COVID-19 patients (P = 0.0016). Additionally, the analysis revealed a remarkable association between rs4646994 SNP and the HB and ESRI levels in patients (P < 0.05). Although the ACE2 rs2285666 SNP was not related to the severity of disease, this variant was significantly associated with ALT, ESRI, and P. These results provide preliminary evidence of a genetic association between the ACE-D/D genotype and the D allele of ACE1 genotype and the disease severity. Therefore, our findings might be useful for identifying the susceptible population groups for COVID-19 therapy.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00438-022-01953-8.

Genetic variation in DPP-IV gene linked to predisposition of T2DM: A case control study.

PurposeDPP-IV is a ubiquitously expressed cell surface protein that can be presented in soluble forms. It has recently gained medical importance as its inhibitors are widely being used as treatment of T2DM. The present research aims to resolve whether genetic variants of DPP-IV have association with susceptibility to T2DM.MethodTwo variants of DPP-IV were detected in 100 controls and 100 T2DM by PCR–RFLP technique. Demographic characteristics were recorded. Clinical characteristics were analyzed by enzymatic method. Statistical analysis was performed using SPSS-21.ResultsDemographic and clinical characteristics differ significantly between two groups. The genetic variation in SNP rs3788979 and SNP rs7608798, both in case and control, were in accordance with Hardy–Weinberg Equilibrium (p value > 0.05). Both SNPs rs3788979 and rs7608798 were significantly related to T2DM (p- < 0.05). Minor G allele of rs3788979 was linked with the susceptibility of T2DM (p-value-0.000; OR- 4.235). T allele of SNP rs7608798 conferred the risk of diabetes with OR-2.235.ConclusionThis is the first attempt to investigate the association of DPP-IV gene with T2DM in Indian population. The finding of study concludes that genetic variation in DPP-IV gene may considerably increase the risk of developing T2DM.

Detection of Novel gyrB Mutation in Fluoroquinolone-Resistant Salmonella and Escherichia coli using PCR-RFLP

Abstract Background Emergence of fluoroquinolone resistance in gut pathogens is a cause of concern. Resistant to quinolone is mainly due to the point mutations at the quinolone-resistance determining regions (QRDR). The aim of the study was to develop polymerase chain reaction-restriction fragment length polymorphism assay (PCR-RFLP) to detect QRDR mutations in gyrA and gyrB regions in enteric pathogens. Methodology PCR-RFLP was done for gyrA 83 region using HinfI and for gyrB 447 using AcuI for fluoroquinolone resistant and susceptible gut pathogens. The products were also sequenced to confirm the presence of restriction sites. Results In this study, a PCR-RFLP technique was developed to detect gyrA 83 mutations in Salmonella typhi and Escherichia coli. A first of its kind PCR-RFLP was also developed to detect gyrB 447 mutation using a restriction enzyme AcuI. Restriction digestion of gyrA using HinfI resulted in three bands for resistant S. typhi isolates due to the presence of mutation at gyrA 83 and four bands were seen for sensitive S. typhi isolates, while two bands for resistant and three bands were seen in sensitive E. coli isolates. Similarly, restriction digestion of gyrB using AcuI resulted in no digestion for resistant S. typhi isolates and two bands for resistant E. coli isolates. This suggest that there is mutation at gyrB 447 region ofE. coli, while no mutation was found in S. typhi isolates. Conclusion The PCR-RFLP developed in the present study could successfully detect gyrA 83 and gyrB 447 mutations in fluoroquinolone-resistant S. typhi and E. coli. The technique can be efficiently used in epidemiological studies instead of a cost-intensive sequencing method to detect the status of multiple point mutations in gut pathogens.

The Effects of BMI and Genetic Variation of Adipokines on Serum Concentrations of Hormones in Untreated Individuals with Breast Cancer; a Pilot Study

Background: Numerous studies have shown an association between hormones secreted by adipose tissues and cancer development. Objectives: This study aimed at investigating the effect of body mass index (BMI) and genetic variation of leptin and adiponectin on serum concentrations of leptin, adiponectin, and estradiol among untreated breast cancer. Methods: This case-control study was performed on 350 women (175 women with breast cancer and 175 healthy controls), who had not taken any medications. Serum levels of estradiol (17-beta estradiol), leptin, and adiponectin were measured, using the ELISA technique. Single-nucleotide polymorphisms of leptin gene (LepG2548A), leptin receptor (Q223, K109R, and K656N), and adiponectin gene (T45G, G276T, C11377G, and 11391A) in blood-isolated DNA were evaluated, using RFLP-PCR technique. Results: Body mass index can affect serum concentrations of hormones and is associated with breast cancer. Also, except for adiponectin C11377G polymorphism, other all genetic variations showed significant relationships with breast cancer. In both groups, BMI was significantly correlated with the mean serum concentrations of hormones, and the risk of breast cancer increased in G2548A, Q223R, K656N, and G276T polymorphisms. The effect of risk allele genotypes on serum concentration of hormones showed that changes in serum concentration of estrogen and leptin in all studied polymorphisms were associated with breast cancer in postmenopausal women. But adiponectin level was only affected by polymorphisms K109R, K656N, and G276 and G11391A. Conclusions: High BMI and genetic variation can affect cancer development by changing the serum concentrations of hormones in different genotypes. Studying various populations’ genetics and lifestyle can help definitive conclusions about genetics and obesity.

Association between Vitamin D Deficiency and Vitamin D Receptor Gene Polymorphism (FokI-rs2228570) among Men with Anemia in Babylon Province, Iraq.

Vitamin D or calciferol is a fat-soluble vitamin that has a unique feature of synthesizing in the body mainly by exposure to UV from the sunlight and then transformed to 25 (OH) D by the liver and finally to a vital form 1,25-dihydroxyvitamin D by the kidneys. Vitamin D receptor gene polymorphism (FokI-rs2228570) has been proposed as the major cause of anemia. The present study aimed to discover the association between vitamin D deficiency and vitamin D receptor gene polymorphism (FokI-rs2228570) in patients with anemia. A total of 120 men with anemia and no kidney disorders have been compared with 60 healthy controls in the present case-control study. A single nucleotide polymorphism (SNP) FokI-rs2228570 was detected by PCR and PCR-RFLP techniques. Levels of serum vitamin D, erythropoietin, and some biochemical parameters were detected by the ELISA assay technique. The mutant homozygous genotype ff was more frequent in patients with anemia (45%) than in the controls (15%). Also, the frequency of the f allele was associated with a significant decrease in the levels of vitamin D and hemoglobin in patients (0.62%); therefore, the mutant allele is a risk factor for developing anemia compared with genetic patterns FF and Ff. Vitamin D deficiency is common in those with anemia which is often associated with low hemoglobin and high levels of erythropoietin. Additionally, the genetic frequencies also affect the level of vitamin D which is indicated by low levels of mutant patterns (Ff, ff) in which patients suffer from severe anemia.

Association of CXCL12 and CXCR4 Genes Polymorphism with Proliferative Diabetic Retinopathy in Diabetic Patients

Introduction: Patients with diabetes are prone to develop complications related to the retina. So far, several genes have been reported in connection with the progression of retinopathy, and a group of these genes is genes involved in angiogenesis. CXCL12 and CXCR4 genes are among the most important genes in this path, which have been less studied and investigated. The aim of this study was to determine the relationship between polymorphism of CXCL12 (rs1801157) and CXCR4 (rs2228014) genes and the risk of proliferative diabetic retinopathy in the population of Yazd City.&#x0D; Methods: In the present case-control study, rs2228014 and rs1801157 polymorphisms were investigated among 103 patients with diabetes in two groups, case and control. In this study, the control group included 49 diabetic patients who did not show retinopathy complications, and the case group consisted of 54 diabetic patients with proliferative retinopathy complications. The Genotyping of rs2228014 polymorphism was examined by ARMS-PCR method and rs1801157 variant by RFLP-PCR technique. The statistical analysis of data was conducted using SPSS software version 16 and chi-square test.&#x0D; Results: Our results represented no association among the rs1801157 and rs2228014 polymorphisms and the risk of proliferative diabetic retinopathy (P&gt;0.05).&#x0D; Conclusion: This study did not confirm association of the rs1801157 and rs2228014 polymorphisms with proliferative diabetic retinopathy and further studies are required to investigate the correlation of polymorphisms of interest with PDR.

The impact of MCP1-2518A/G and CCR2-V64I genetic polymorphisms in Egyptian sickle cell disease patients

BackgroundSickle cell disease (SCD) is an inherited genetic disorders of hemoglobin that causes multisystem morbidity. The pathophysiology of SCD is complex and includes HbS polymerization/sickling, hemolysis, endothelial dysfunction and inflammation. Chemokines are proteins playing an important role in the inflammation process and could be involved the context of pro-inflammatory SCD. Some chemokine polymorphism were found to be associated with clinical complication in SCD. Aim of the studyWas to explore the frequency and the possible effect of Monocyte Chemo-attractant Protein 1–2518A/G (MCP1–2518A/G) and Chemokine Receptor 2-V64I (CCR2-V64I) genetic polymorphisms on clinical and laboratory disease-related variables in Egyptian Sickle cell disease patients. Patients and methodsGenotyping of the two genes were performed by PCR-RFLP technique for 80 SCD patients as well as 50 healthy control group. ResultsThe study revealed that the MCP1–2518 polymorphic genotypes (AG & GG) showed no statistically significant difference in the distribution of the polymorphic genotypes between SCD patients and the controls (p = 0.164). While a significantly higher frequency of the mutant variants CCR2-V64I GA/AA among SCD patients than the control subjects were found (p = 0.032). Regarding the clinic-pathological features, the frequency of recurrent infections, vaso-occlusive crisis, severe vaso-occlusive crisis and number of hospitalization/year were higher in patients harbouring the MCP1–2518A/G and CCR2-V64I polymorphic genotypes than the wild genotype, and gall bladder complications were higher in MCP1–2518 G allele patients, whereas surgical splenectomy were higher in CCR2-V64I A allele patients (p < 0.05). In conclusionMCP1–2518A/G and CCR2-V64I genetic polymorphisms may influence the clinical severity of sickle cell disease.

TLR3 polymorphisms in HIV infected individuals naïve to ART.

TLR3 polymorphisms affect the risk of HIV infection and modify the disease course. Consequently, we analyzed the association of TLR3 polymorphism (rs5743312, rs3775296, and rs3775291) with susceptilbity to HIV-1 acquisition and disease progression. This is a cross-sectional study. Genotyping of TLR3 polymorphisms were completed by the utilization of the PCR-RFLP technique in 153 HIV naive subjects and 158 healthy controls. A haplotype is a physical grouping of genomic variants that tend to be inherited together. The TCC haplotype was increased in HIV infected individuals compared with healthy controls (0.05% versus 0.03%). TLR3 rs3775291CT genotype was associated to the early stage of HIV infection (OR=2.19, P=0.04), with a higher occurrence in advance stage of HIV infection when contrasted with healthy controls (41.2% versus 32.3%). TLR3 rs3775296 CA genotype was likely to be associated with intermediate stage of HIV infection (19.5% versus 31.6%, OR=0.42, P=0.06). TLR3 rs5743312TT genotype was used to be greater prevalence in advanced stage of HIV infection compared with healthy controls (2.9% versus 1.9%). TLR3 rs3775296CA genotype was less prevalent in HIV subjects devouring tobacco when contrasted with non-users (9.1% versus 34.9%, OR=0.25, P=0.09). TLR3 rs3775296AA and rs3775291CT and TT genotypes have been overrepresented in HIV subjects using alcohol when contrasted with non-users (5.6% versus 1.1%, OR=1.83, P=0.67; 50.0% versus 42.2%, OR=1.84, P=0.31; 5.6% versus 3.3%, OR=2.70, P=0.50). In multivariate examination, rs5743312TT genotype showed a greater risk for HIV infection (OR=1.86, P=0.50). TLR3 rs3775291 C/T polymorphism may assist the risk of disease progression in alcohol consumers. TLR3 rs3775291 CT genotype may enhance the disease progression whereas the TLR3 rs3775296 CA genotype may protect for disease progression.

Polymorphisms of miR-146a and susceptibility to ulcerative colitis risk: a case-control study

Considering the role of miR-146a in the control of inflammation, we assessed the importance of two miR-146a polymorphisms (rs2910164 and rs57095329) in the development and severity of ulcerative colitis (UC) in Iran. Genomic DNA of 150 cases with UC and 200 healthy individuals were genotyped using the PCR-RFLP technique. Statistical analyses were performed using Med Calc software. The miR-146a rs2910164 C allele was significantly associated with increased risk of UC. Individuals carrying the CC (rs2910164) were more than fourfold higher risk of UC relative to wild type homozygotes. The combined GC + CC genotypes were also associated with increased UC risk. We also found that the rs2910164 CC genotype was associated with a severe form of the disease However, the distribution of variant allele and genotypes of rs57095329 did not differ between the cases and controls. In conclusion, miR-146a rs2910164 polymorphism may play a role in UC. To confirm our findings, additional well-designed studies in diverse ethnic populations are required.

The role of FASL, BCL-2 and BAX polymorphisms in brazilian patients with prostate cancer and benign prostatic hyperplasia.

Prostate cancer (PCA) is one of the leading causes of death among men, being related to several factors, including the aging male population, like benign prostatic hyperplasia (BPH), a histopathological and hyperplastic alteration associated to prostate aging. The FASL, BCL-2 and BAX genes are involved in cell apoptosis regulation and can be related to the development of both cancer and hyperplasia. This study aimed to investigate the association of FASL - 844 (rs763110), BCL-2 -938 (rs2279115) and BAX - 248 (rs4645878) polymorphic variants in Southern Brazilian PCA and BPH patients and healthy controls. 348 samples were analyzed, being 123 from PCA patients, 143 BPH patients and 82 healthy controls, using PCR-RFLP techniques. The results of genotyping analysis were adjusted by age, and compared with PSA levels and prostate volume. The analyzes of genotype frequencies according to PCA, HPB and controls, were performed by logistic regression corrected by age, and showed that the FASL CC genotype can be a risk factor for PCA patients, when compared to controls (p = 0.041). The clinical data investigation indicated higher PSA levels in PCA patients with FASL CC genotype, as compared to TC genotype carriers (p = 0.044), higher PSA levels for healthy individuals with BCL-2 AA genotype, comparing with CC genotype (p = 0.027) and higher PSA levels in BPH group with FASL CC genotype, as compared to TC genotype (p = 0.044). Our data indicate the FASLCC genotype as a risk factor for prostate pathologies, whileBCL-2 CC can act as a protective genotype.

Genotyping of FOXO3a gene polymorphism rs13217795 (C &gt; T) in asthmatic Iraqi patients

Background: Asthma is a chronic inflammatory process that affects the airways of the lung. It has a greater impact on public health. It is a polygenic disease, many factors play a role implicated in triggering its that starts like genetic, environmental, and geographic factors. Genetic studies have focused on the forkhead box O3 (FOXO3a) gene, mainly polymorphism at the specific sites which is thought may contribute to exacerbating the attack and complications of asthma. Methods: The study was designed as a case-control and involved 138 of asthmatic patients and 120 seemingly healthy people used as a control group. The diagnosis was carried out by consultants at a special respiratory center in Babylon governorate. Genotyping of all subjects was done for FOXO3a (rs13217795; C/T) analysis by PCR-RFLP technique. Multinominal regression was used to calculate Odds ratios (ORs) and 95% confidence intervals (CIs) by IBM. SPSS software version 20.0. Results: Analysis of the results that obtained form this work a considerable modification in forms of rs13217795; C/T polymorphism between asthmatic and healthy individuals. elevation in the risk of asmathic patient was seen in T allele of FOXO3a gene (odds ratio of 2.0313, 95% CI: 1.5493 - 2.6631, P&lt;0.0001).

Association of TNFSF11 rs2200287 and TNFSF11 rs2148072 gene polymorphisms in preeclampsia.

Various cytokines released by white blood cells like lymphocytes are linked to the immune system during pregnancy. The polymorphism of the TNFSF11 (rs2200287 and rs2148072) gene is related to the preeclampsia of pregnancy. This study was a prospective study involving 304 pregnant women with preeclampsia (n=152) and controls (non-preeclamptic pregnant women) (n=152). To investigate the rs2200287 and rs2148072 SNP's of TNFSF11 gene polymorphism by using the PCR-RFLP techniques. A significantly different genotype distribution of TNFSF11 (rs2200287 and rs2148072) polymorphisms were observed between the two groups, with the G allele of variant rs2200287 was highly significant in the preeclamptic group (P=0.000; .5814; OR=.5814; 95% CI=.4211-.8012). And the C allele of variant rs2148072 was also highly significant in the preeclamptic group (P=0.000; OR=.5076; 95% CI, .362-.71) in this study. The outcomes of the present study indicate that there was an association in TNFSF 11 (rs2200287 and rs2148072) gene polymorphism with preeclampsia compared to non-preeclampsia women.

A PCR-RFLP Technique to Assess the Geographic Origin of Plasmodium falciparum Strains in Central America

The elimination of malaria requires strengthening diagnosis and offering adequate and timely treatment. Imported cases of falciparum malaria represent a major challenge for pre-elimination areas, such as Central America, where chloroquine and primaquine continue to be used as first-line treatment. The pfs47 gene has been previously described as a precise molecular marker to track the geographic origin of the parasite. The aim of this study was to design a simple and low-cost technique using the polymorphic region of pfs47 to assess the geographic origin of P. falciparum strains. A PCR-RFLP technique was developed and evaluated using the MseI enzyme that proved capable of discriminating, with reasonable precision, the geographical origin of the parasites. This method could be used by national surveillance laboratories and malaria elimination programs in countries such as Honduras and Nicaragua in cases of malaria where an origin outside the Central American isthmus is suspected.