Reaction-restriction Fragment Length Polymorphism Research Articles

Cyclin D1 (G870A) polymorphism and breast cancer risk in an Iranian population

Background and Objective: Cyclins are the key regulator of the cell cycle and their over-expression has been seen in many cancers including breast cancer. Cyclin D1 is an oncoprotein encoded by CCND1 gene located on chromosome 11 (11q) which regulates cell cycle in shifting from G1 to S phase. It’s the main target for steroids and mitogenic growth hormones in breast epithelial cells. This study aimed to evaluate the relationship between Cyclin D1 G870A polymorphism and breast cancer risk in a population in the north of Iran.
 Methods: Whole blood samples collected from 82 patients with breast cancer and 66 healthy women. DNA was extracted and genotyping was performed by Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) technique.
 Results: Genotypic prevalence of AA, AG, GG genotypes among patients were 40.2%, 35.3% and 24.4% and in controls were 30%, 47%, 23%, respectively. There was no significant difference in CCND1 G870A genotype polymorphism between patients and control group (p=0.32). Also, allelic prevalence of A and G alleles in breast cancer patients were 58% and 42%, in controls were 54% and 46%, respectively. The present study showed that there is no significant association between CCND1 G870A polymorphism with the risk of breast cancer.
 Conclusion: The results of this study revealed that there is no significant association between CCND1 G870A genetic polymorphism and the risk of breast cancer in the population of the north of Iran. More studies with larger samples of cases and controls would be beneficial.

Association between Vitamin D Receptor Polymorphism and Susceptibility to Oral Lichen Planus and Oral Squamous Cell Carcinoma.

Oral squamous cell carcinomas (OSCC) comprise 90-95% of oral cancers. Early diagnosis improved the survival rate of OSCC patients to 80-90%. Oral lichen planus (OLP) is a chorionic inflammatory disease with malignancy potential. The vitamin D receptor (VDR) plays a critical role in the pathogenesis of oral cancer. This study aimed to determine the association between VDR rs7975232 (Apa I) polymorphism and potential susceptibility to OLP and OSCC risks. In this prospective case-control study, a total of 120 blood samples were obtained from OSCC patients (n=29), OLP (n=50), and controls (n=40). VDR rs7975232 polymorphism was studied using the Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) method. Statistical analysis was performed with SPSS Version 23 software. Data were expressed as means ± standard deviation (SD). Age, sex, allelic frequency, and genotyping were compared using the chi-square test. A p-value of less than 0.05 was regarded as statistically significant. The disease risk was estimated by Odds ratio (OR) with a 95% confidence interval. A significant age difference was observed between the controls and the OSCC group (p=0.001). A significant difference was observed in Aa and aa genotypes compared with AA between OSCCs and controls. Moreover, dominant (p<0.001), additive (p<0.001), and allelic (p=0.001) models were different between groups. There was a positive association between rs7975232 VDR polymorphism and susceptibility to OSCC. More experimental evidence must reveal the possible association between rs7975232 and the risk of OLP in a larger cohort.

Genetic Variations of Angiotensinogen, Angiotensin Converting Enzyme, and Angiotensin Type 1 Receptor with the Risk of Pulmonary Tuberculosis.

There is little data regarding the impact of renin-angiotensin system (RAS) gene polymorphisms on tuberculosis. The current study designed to survey the possible association between RAS polymorphisms and the risk of pulmonary tuberculosis (PTB) in a sample of the southeast Iranian population. This case-control study was done on 170 PTB patients and 170 healthy subjects. The AGT rs699 C&gt;T, ACE rs4341 C&gt;G and AT1R rs5186 C&gt;A variants were genotyped using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and ACE rs4646994 (287bp I/D) variant by PCR method. Regarding AT1R rs5186 A&gt;C polymorphism, the findings revealed that AC genotype and C allele significantly decreased the risk of PTB (OR=0.39, 95% CI=0.22-0.67, p=0.001, and OR=0.53, 95% CI=0.25-0.72, p=0.002, C vs. A, respectively). The TC genotype and C allele of AGT rs699 T&gt;C significantly associated with decreased the risk of PTB (OR=0.45, 95% CI=0.28-0.74, p=0.002, TC vs. TT and OR=0.51, 95% CI=0.32-0.80, p=0.005, C vs. T, respectively). The ID genotype of ACE 287bp I/D significantly increased the risk of PTB (OR=1.88, 95% CI=1.12-3.17, p=0.017). Our finding did not support an association between ACE rs4341 C&gt;G variant and the risk of PTB. In summary, the findings revealed an association between AT1R rs5186 A&gt;C, AGT rs699 T&gt;C and ACE 287bp I/D polymorphisms and the risk of PTB in a sample of the southeast Iranian population. Further investigation with higher sample sizes and diverse ethnicities are required to confirm our findings.

The Association between Histidine-Rich Glycoprotein rs10770 Genotype and Recurrent Miscarriage in Iranian Women.

Recurrent miscarriage (RM) is a significant reproductive concern affecting numerous women globally. Genetic factors are believed to play a crucial role in RM, making the histidine-rich glycoprotein (HRG) gene, a topic of interest due to its potential involvement in angiogenesis. This study is aimed at investigating the association between the HRG rs10770 genotype and RM. Blood samples were collected from a total of 200 women at the beginning of the study. Subsequently, a comparative analysis was conducted between the blood samples of 100 women with a history of RM (case group) and the blood samples of another 100 healthy women (control group). HRG rs10770 genotyping was performed through polymerase chain reaction restriction-fragment length polymorphism (PCR-RFLP), followed by statistical analysis to evaluate the relationship between HRG rs10770 genotype and RM. The results indicated a significant statistical difference between the C/C genotype (OR = 3.32, CI: 1.22-9.04, p = 0.01) and the C/T genotype (OR = 1.24, CI: 0.67-2.30, p = 0.47) in both the case and control groups. Additionally, a significant correlation was observed in the C allelic frequency among RM participants compared to the control group (OR = 1.65, CI: 1.06-2.58, p = 0.02). The study highlights the importance of HRG rs10770 in understanding RM, shedding light on its implications for reproductive health. Furthermore, it became evident that women carrying the homozygous C/C genotype exhibited increased susceptibility to the risk of RM.

Genetic Risk of Rheumatoid Arthritis: A Case Control Study.

Vitamin D effects are mediated by vitamin D receptors (VDRs), which are influenced by various genetic polymorphisms, including ApaI and BsmI. These polymorphisms have been linked to several diseases, including rheumatoid arthritis (RA). This study aimed to compare the frequency and association of VDR ApaI and BsmI gene polymorphisms, serum 25-hydroxy vitamin D (25-(OH)-D) levels, and calcium (Ca) levels between a RA group and a matched healthy control group. In one hundred RA patients and fifty healthy controls, the genotypes of the VDR ApaI and BsmI gene polymorphisms were analyzed using polymerase chain reaction restriction fragment length polymorphisms (PCR-RFLP). Both Serum 25-(OH)-D level and calcium level were measured in the two groups. There was no significant difference between the cases and controls regarding the VDR ApaI gene polymorphism (p = 0.89). A significant difference was observed between the cases and controls in terms of the VDR BsmI gene polymorphism (p = < 0.001). The serum levels of 25-(OH)-D and calcium were significantly lower in the RA group compared to the control group (p = 0.04 and < 0.001 respectively). Significantly higher serum vitamin D levels were associated with the aa genotype (p = 0.007). Significantly increased calcium levels were associated with the AA genotype (p = 0.02). No significant difference was found among BsmI polymorphisms regarding vitamin D and Ca levels (p = 0.25 and 0.87 respectively). Vitamin D receptor gene BsmI polymorphism but not ApaI polymorphism could be a marker of RA susceptibility. Vitamin D and Ca levels are negatively affected by RA. Vitamin D receptor gene ApaI polymorphism contributes to vitamin D and Ca levels.

Study the Effect of RAGE and HMGB1 Genes Polymorphism on Breast Cancer Susceptibility in Iraqi Female

Background: The most frequent malignancy among women globally is breast cancer (BC). Breast lobules or ducts are the genesis of breast cancer, which is an uncontrolled development of epithelial cells. High mobility group protein box 1 (HMGB1) possesses both pro- and anti-tumorigenic bioactivities, which contribute to its complicated function in carcinogenesis. Damage associated Molecular Pattern molecules (DAMPs) generated following tissue injury include HMGB1, many S100 proteins, and others. RAGE is thought to be a receptor for these moleculAim of the study: The study was aimed to find the High mobility group protein box 1 (HMGB1) HMGB1 (rs1412125) and Receptor for Advanced Glycation End Products RAGE (rs1800624) genotypic and allelic frequency. Using Allele Specefic polymerase chain reaction AC-PCR and Polymerase Chain Reaction Restriction Fragment Length Polymorphism RFLP-PCR in BC patients and healthy controls, as well as their correlation with breast cancer susceptibility.Material and methods :. Samples were taken at Marjan Cancer Hospital, Babylon. HMGB1 rs1412125 and RAGE rs18400624 polymorphisms were evaluated for patients and controls using the PCR-RFLP and allele-specific polymerase chain reaction (AS-PCR) method. SPSS software was used to perform the statistical analysis.Results : There is non-significant difference between patients and control in TT, AA, and TA genotyping of (rs1800624) in RAGE gene (p=0.22), non-significant difference between patients and control in TT, TC, and CC genotyping of (rs1412125) in HMGB1 gene (p=0.09).Conclusion : Our results showed no association between HMGB1 polymorphisms and RAGE and breast cancer risk.

Genetic diversity of Toxoplasma gondii in goats and sheep from the Northeast Region of Brazil destined for human consumption

This study aimed to genotype isolates of Toxoplasma gondii obtained from samples of brain, diaphragm and heart of goats and sheep intended for human consumption in the State of Paraíba, Brazil. Tissue samples from 14 animals, goats (n = 5) and lambs (n = 9), were sourced from public slaughterhouses in seven cities and bio-assayed in mice. The brains of the mice were utilized for DNA extraction. Genotyping was carried out by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) using 10 markers (SAG1, SAG2, SAG3, BTUB, c22-8, PK1, GRA6, L358, c-29-2 and Apico). A total of 10 isolates were fully genotyped (i.e. at all loci), three from goats and seven from sheep, revealing five distinct genotypes: #13 (n = 4); #48 (n = 3); #57 (n = 1); #273 (n = 1); and one new genotype that had not been previously described. Genotype #13 is frequently found in the Northeast of Brazil and represents a clonal lineage circulating in this region and was the most prevalent genotype identified (n = 4). Moreover, in the present study genotypes #13, #48, #57, and #273 were documented for the first time in sheep from Brazil, and the novel genotype was isolated from a goat. Our findings align with previous studies on T. gondii from Brazil, where new genotypes are continuously being identified, highlighting a high level of genetic diversity of T. gondii isolates in the country.

Macrophage Migration Inhibitory Factor and Gene Polymorphism (rs755622G&gt;C) in Unstable Vitiligo Patients

Vitiligo pathogenesis is related to the macrophage migration inhibitory factor (MIF) protein. This study aimed to assess the lesional MIF levels and gene polymorphisms (rs755622G&gt;C) in patients with vitiligo. To assess the consequences of combining narrow-band ultraviolet B with oral minipulse prednisolone as opposed to a combination with oral methotrexate on MIF levels in vitiligo patients, 50 unstable vitiligo patients and 50 controls were randomly chosen for comparison. MIF levels in skin homogenates and MIF (rs755622G&gt;C) single nucleotide polymorphisms were assessed using the ELISA and the polymerase chain reaction restriction fragment length polymorphism (RFLP-PCR) techniques. We found significantly higher lesional MIF levels, a higher frequency of both (GC) and (CC) genotypes, and a significantly more frequent mutant allele (C) in patients than in controls. In addition, there was a significantly lower frequency of the allele (C) among patients who exhibited moderate to marked therapeutic improvement than among those who showed minimal to mild improvement. In conclusion, tissue MIF and gene polymorphisms were associated with vitiligo. In addition, oral corticosteroids, narrow-band ultraviolet B, methotrexate-targeted tissue MIF, and gene polymorphisms can improve unstable vitiligo.

Genetic polymorphism of interleukins 6 and 17 correlated with apical periodontitis: A Cross-sectional study.

Interleukins 6 and 17 act in bone resorption in the presence of infections of endodontic origin for host defense. Genetic polymorphisms may be associated with increased bone loss, represented by areas of large periapical lesions. This study aimed to verify the frequency of interleukin 6 and 17 gene polymorphism in patients with asymptomatic apical periodontitis or chronic apical abscess and to verify the existence of correlations between periapical lesion area with age, gender, and presence of the polymorphism, in the studied population, in the state of Pernambuco. A population consisting of thirty diagnosed individuals was included. The area of the lesions was measured in mm². Genomic DNA was extracted and genotyping was performed by Polymerase Chain Reaction Restriction Fragment Length Polymorphism for interleukin 6 (rs 1800795) and interleukin 17 (rs 2275913). Fisher's exact, chi-square, and odds ratio tests were used. A logistic regression analysis was also performed using sex, age, and the presence of polymorphism as covariates, in addition to linear regression to test the relationship between age and lesion area. All tests used a significance level of 0.05% (p ≤0.05%). There was no statistical significance in the occurrence of large areas of periapical lesions correlated with age, sex, and diagnosis, nor in the distribution of alleles in the polymorphism of interleukins 6 and 17 in the studied groups. The frequency of homozygous and heterozygous polymorphism was high. The polymorphism of these interleukins is not correlated with the increase in the areas of asymptomatic periapical inflammatory lesions.

Association of NFKB1 gene polymorphism (rs28362491) with cardiometabolic risk factor in patients undergoing coronary angiography.

Genetic and environmental factors are involved in the pathogenesis of cardiovascular diseases (CVDs). The aim of the study was to investigate between the genotype of the NFKB1 gene and the cardiometabolic risk factor in patients undergoing coronary angiography. This cross-sectional study was conducted on 462 adults (male and women) aged between 35 and 75 years who referred to Afshar Hospital for coronary angiography in 2021- 2022. The polymerase chain reaction restriction fragment length polymorphism method was used to detect the genotype of rs28362491. Biochemical parameters were measured using commercial kits. Gensini and Syntax scores were calculated using the angiography result to assess the extent of coronary artery stenosis. We used multivariate logistic regression analysis to examine the relationship between genotype variants and cardiometabolic risk factors. There was no association between variant genotypes and abnormally levels of serum alanine aminotransferase (ALT) (P value=0.51), aspartate aminotransferase (AST) (P value=0.99), triglyceride (TG) (P value=0.48), total cholesterol (P value=0.79), low density lipoprotein-cholestero (LDL-C) (P value=0.31), high-density lipoprotein-cholesterol (HDL-C) (P value=0.53), fast blood sugar (FBS) (P value=0.39), systolic blood pressure (P value=0.14), diastolic blood pressure (P value=0.64), Gensini score (P value=0.48) and syntax score (P value=0.74) in the crude model even after adjustment for confounding factors. We found no association between the ATTG polymorphism and cardiometabolic risk factors in patients who had coronary angiography. Further investigations are needed to assess the association between variants of 28362491 and cardiometabolic markers.

IL-10 (−1082 G/A) polymorphism in Bataknese with schizophrenia

ObjectivesThree biallelic polymorphisms at the promoter region of the interleukin-10 (IL-10) gene have been associated with susceptibility to schizophrenia. The aim of this case-control study was to investigate the association between IL-10 (−1082) G/A gene polymorphisms and schizophrenia among Bataknese, a native tribe inhabiting the North Sumatera province in Indonesia. MethodsA total of 194 unrelated participants (n = 97 for each case and control groups) participated in this study. Polymerase chain reaction restriction fragment length polymorphism molecular genotyping was conducted to assess the genotype and allele distribution of IL-10 (−1082 G/A). ResultsAllele variations indicated that the dominant allele in the Batak tribe was allele A, whereas homozygous GG genotypes were not found in either group. The A allele and AA genotype were found to be risk factors for developing schizophrenia (OR = 2.26, 95% CI = 1.1825–4.3559 and OR = 2.56, 95% CI = 1.280–5.152, respectively). ConclusionOnly the A allele and AA genotype of the IL-10 gene polymorphism at −1082 G/A contribute to schizophrenia susceptibility in Bataknese.

Investigation of the Association Between ITPA Gene 94C>A Sequence Variant and Kidney Transplant Rejection in Iranian Kidney Transplant Recipients.

Thiopurine prodrugs are commonly used in kidney transplant recipients. Inosine triphosphate pyrophosphatase is an enzyme encoded by the ITPA gene. Alteration of ITPA gene is one of the pharmacogenetic sequence variants possibly involved in thiopurine metabolism. The ITPA 94C>A sequence variant (C-to-A substitution at nucleotide 94) is associated with an increased risk of adverse drug reactions in patients treated with the thiopurine drug. The aim of the present study was to investigate the effect of the ITPA 94C>A gene sequence variant in kidney transplant recipients. The genotyping of the ITPA rs1127354 variant was performed by the polymerase chain reaction restriction fragment length polymorphism method in 140 kidney transplant recipients and in 100 control participants. Data were analyzed with SPSS statistical software. The results revealed a significant difference between control and nonrejection groups regarding the rs1127354 genotype and allele frequency. No significant difference was found between the rejection and nonrejection groups regarding the rs1127354 genotype and allele frequency. Also, a significant association was observed between the ageofthe control group and age of the rejection group. No significant differences between sex and underlying disease in patients with or without rejection were observed. We observed no significant differences between rejection and nonrejection transplant. Further studies are recommended, in a larger population and with different ethnicities.

Relationship between polymorphisms in the CD40 gene with the prevalence of breast cancer: A case-control study

Background. Breast cancer with a complex inheritance pattern is a major cause of cancer death among women worldwide. Single nucleotide polymorphisms (SNPs), the most common genetic variations, influence interindividual predisposition to disease and treatment outcomes with drugs. Evidence suggests that CD40 polymorphism contributes to pathogenesis of cancer. The co-stimulatory molecule CD40 plays a prominent role in immune regulation. This study aimed to test the association between polymorphisms in the CD40 gene and breast carcinogenesis in Arak, Iran. Methods. In this case-control study, three SNPs (rs1883832, rs4810485, rs3765459) were genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. We included 80 patients with breast cancer and 80 healthy controls. Statistical analysis was performed by SPSS (version 26) using Chi-Squared test at P˂ 0.05. Results. Our data showed a statistically significant association between the two CD40 SNPs (1s1883832 and rs4810485) and breast cancer risk (P=0.038 and P=0.000, respectively). There was no significant association between rs3765459 and breast cancer risk (P=0.190). Conclusion. We witnessed that CD40 gene polymorphisms (rs1883832 and rs4810485) contributed to breast cancer. So, they are associated with breast cancer risk. Practical Implications. The obtained data revealed a significant relationship between the rs1883832 and rs4810485 polymorphisms and the risk of breast cancer. Thus, these polymorphisms could be used as biomarkers to predict breast cancer.

Association of IL4RA polymorphism in predicting susceptibility towardchronic obstructive pulmonary disease.

Background: The cytokine IL-4 plays vital role in the intercellular signalling network during immune responses to allergen exposure. Methods: This cross-sectional study involved 202 chronic obstructive pulmonary disease (COPD) patientsand 203 healthy individuals. The genotyping of IL4RAQ576R gene polymorphism was determined using PCR restriction fragment length polymorphism analysis. Results: Significant association between mutant genotype (GG) and combined (AA+AG) genotype for the risk of COPD was found (odds ratio [OR]: 4.32;p=0.04). A significant protective effect was observed between the IL4RAQ576R polymorphism and Global Strategy for Obstructive Lung Disease (GOLD) stage fourpatients in a recessive model (AA+AG vs GG; p=0.002). In GOLD A, asubstantial relationship was found between theAG and wild-type genotypes (AA) for COPD risk (OR: 2.38;p=0.03). A strong association was found for COPD duration of 5-10years (OR: 8.80;p=0.01). Conclusion: IL4RAQ576R polymorphism is associated with COPD susceptibility.

Study of the Association of Vitamin D Receptor (VDR) Gene Polymorphism in Iraqi Patients with Inflammatory Bowel Disease

Inflammatory bowel disease (IBD) is a chronic inflammatory disorder divided into two types, Ulcerative colitis (UC), and Crohnʹs disease (CD). In total, 80 blood samples were collected for this study, 40 blood samples were collected from IBD and 40 blood samples were collected from healthy individuals as. Genomic DNA extracted from whole blood and (FokI) polymorphism was detected by polymerase chain reaction Restriction fragment length polymorphism (PCR- RFLP). The results of the statistical analysis showed high frequency and percentage in age group (&lt; 30 years) was 16 (40%) with significant (P≤0.05) when compeered with control group. The frequency and percentage of body mass index (BMI) for the patient group was (27.91 ± 0.45). While BMI was (27.31 ± 0.54) for the control group with highly significant association (p&lt;0.01) between Iraqi patients with IBD when compeered with control group.(VDR- FokI) wild allele PCR products (without restriction site) have 265bp (F allele) and when digested and electrophoresis of the digestion products on agarose gel the resulted fragment size of 196bp, 265 bp and 69 bp (f allele).The genotype of FokI gene (TT/FF) was 14 (35%) (P-value=0.0039), TG / Ff 15(37.5%) (p= 0.0253), GG /ff 11(27.5%) (p=0.0039) in patient group which have a highly significant in frequency and percentage, as compared with control group. In conclusion The FokI genes polymorphism was shown to be associated with an increased incidence of Iraqi patients with IBD when compeered with control group. The genetic factors of FokI gene polymorphism may have a role in inflammatory bowel disease.

Identification polymorphism of LHR and FSHR genes in Indonesian Holstein dairy cattle associated with productive and reproductive traits

Abstract. Setyorini YW, Kurnianto E, Sutopo, Sutiyono. 2023. Identification polymorphism of LHR and FSHR genes in Indonesian Holstein dairy cattle associated with productive and reproductive traits. Biodiversitas 24: 2898-2905. Luteinizing and follicle-stimulating hormones and their receptors play an important role in the reproductive system’s hormonal activity and physiological function. This study aimed to identify the genotype variation of the Luteinizing Hormone Receptor (LHR) gene exon 11 and the Follicle-stimulating Hormone Receptor (FSHR) gene exon 10 and to elucidate the associating between polymorphism with the milk and reproductive traits in dairy cattle. A total of 100 samples of deoxyribonucleic acid (DNA) from Indonesian Holstein dairy cattle were used in this study. Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) was performed for genotyping of the Single Nucleotide Polymorphism (SNP) of the LHR and FSHR genes with the HhaI and AluI restriction enzymes, respectively. The collected phenotype data were milk production, quality, and reproductive trait. In addition, the general linear model procedure in the SAS program was carried out to investigate the association of genotypes with productive and reproductive traits. The results showed that the LHR gene in the population of this study was monomorphic, while the FSHR gene was polymorphic with three genotypes, namely CC, CG, and GG. The sequences of the FSHR gene indicated the presence of a mutation at nucleotide number 2037 that substitutes cytosine for guanine. Cows with the CC genotype showed better parameter service per conception (P&lt;0.05) than the other genotypes but did not correlate with other reproductive parameters and milk traits. Therefore, it was concluded that SNP g.2037C&gt;G polymorphism at FSHR gene exon 10 was identified in the Indonesian Holstein Dairy cattle population and associated with service per conception parameters but not with milk trait.

Genetic heterogeneity of thalassemia major patients in Rembang Regency, Central Java, Indonesia

Link of Video Abstract: https://www.youtube.com/watch?v=RG95aJkUndg Background: Thalassemia and hemoglobinopathies are caused by mutations in the globin genes that reduce haemoglobin synthesis. In many Asian countries, including Indonesia, thalassemia is a common inherited disorder. Thalassemia has specific characteristics due to the wide variability of the mutations from various populations. Therefore, screening protocol should be designed on specific populations’ regional gene mutation patterns. This study aims to investigate the mutational diversity of the globin genes in thalassemia major patients in Rembang Regency, Central Java, Indonesia. Methods: Genetic mutations were carried out in all major thalassemia patients who were recorded and underwent routine blood transfusions at the Rembang District Hospital from 2018 – 2020. Mutation identification was carried out using a PCR- RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) based method, the ACRS (Amplified Created Restriction Site) method, and genome sequencing. Results: The most common genotypes are double heterozygous HbE and Cd 35 mutation (-C) (42.87%), HbE and IVS1-nt.5 mutations (G&gt; C) (28.57%), HbE and IVS1- nt.2 mutations (T&gt; C), HbE and codon 8/9 (insertion + G), codon 8/9 (insertion + G) and Hb Malay, as well as Hb Adana and Hb Constant Spring (1.74%). Conclusion: The β-thalassemia and α-thalassemia mutations most commonly found in Rembang Regency are Cd 35 (-C) and non-deletional mutations.

A Pilot Study on the Prevalence of Transcription Factor 7-Like 2 Gene (TCFL2), Rs290487 in Ethnic Groups with Type 2 Diabetes Mellitus in Bayelsa State of Nigeria

The negative impact of type 2 diabetes mellitus (T2DM) on individuals, families, health system and the economic development as a whole, is an important justification for research in this field. The present study is a hospital-based case-control type to ascertain the genetic susceptibility of T2DM among a sample population of various ethnic groups resident in Bayelsa State, Nigeria. Also evaluated was the relationship between transcription factor 7-like 2 (TCFL2) rs12255372 and rs290487 genetic polymorphisms with development of T2DM. Genotyping of TCFL2 rs12255372 and rs290487 were carried out using the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. Among the indigenous groups, the allelic frequencies determined for the abnormal (CC) and the normal (TT) genotypes were 94.4 and 5.6 (%), respectively; whereas, the values 91.7 and 8.3 (%), in that other, were determined for the non-indigenous groups. A convincing significant difference was found between subjects with T2DMne and their counterpart controls within indigenous people of Bayelsa State with respect to confounders, including age (t=8.046, p&lt;0.00001), BMI (t =2.628, p&lt;0.0190), waist circumference (t=2.800, p=0.0091) and fasting blood sugar (t=3.212, p&lt; 0.0006). This study verified the association of TCFL2 rs290487 with the development of T2DM in indigenous and non-indigenous people in Bayelsa State. The risk conferred by the homozygous CC genotype was higher than that of the heterozygous TC state – a fact indicative of an additive model of inheritance.

The association between PTPN22 C1858T gene polymorphism and type 1 diabetes mellitus: an Indonesian study

Background Type 1 diabetes mellitus (T1DM) is disease caused by the destruction of β pancreatic cells. The activation of T-lymphocyte and proliferation inhibitor are induced by protein tyrosine phosphatase non-receptor type 22 (PTPN22). However, the link between PTPN22 C1858T gene polymorphism and T1DM is still controversy. This study aimed to analyse the C1858T gene polymorphism in Indonesian children with T1DM. Materials and methods This case-control study was conducted from March 2021 to May 2022 in the Endocrinology Outpatient Clinic at Dr. Soetomo Hospital and Tropical Disease Center Universitas Airlangga. Patients with controlled T1DM during the study period were included. The PTPN22 analysis used polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. Results Sixty-two children voluntarily participated in this study, and were equally divided into the T1DM and control groups. Most of the patients (94%, 58/62) are Javanese. This study revealed a more frequent CC genotype (9.4%) and allele-C (54.6%) polymorphism in the T1DM group, while more frequent CT genotype (100%) and allele-T (50%) polymorphism were in the control group. The C- and T-allele frequency was 54.6% and 45.4% in the T1DM group, respectively. The T1DM and control groups did not significantly differ (p= .2381). Conclusions PTPN22 homozygous genotype-CC and allele-C polymorphisms are more frequent in patients with T1DM. However, the PTPN22 C1858T gene polymorphism did not significantly correlate to T1DM children in this study. Key Messages: The PTPN22 C1858T gene polymorphism does not significantly affect the susceptibility of T1DM in Indonesian children. PTPN22 homozygous genotype-CC polymorphism was more observed in the T1DM group; thus, this genotype may play as a risk factor for T1DM children in the Indonesian population.

Molecular Detection of Phytoplasmas of the 16SrⅠ and 16SrXXXⅡ Groups in Elaeocarpus sylvestris Trees with Decline Disease in Jeju Island, South Korea

Phytoplasmas were discovered in diseased Elaeocarpus sylvestris trees growing on Jeju Island that showed symptoms of yellowing and darkening in the leaves. Leaf samples from 14 symptomatic plants in Jeju-si and Seogwipo-si were collected and phytoplasma 16S rRNA was successfully amplified by nested polymerase chain reaction using universal primers. The sequence analysis detected two phytoplasmas, which showed 99.5% identity to 'Candidatus Phytoplasma asteris' and 'Ca. P. malaysianum' affiliated to 16SrI and 16SrXXXII groups, respectively. Through polymerase chain reaction-restriction fragment length polymorphism (RFLP) analyses using the AfaI (RsaI) restriction enzyme, the presence of two phytoplasmas strains as well as cases of mixed infection of these strains was detected. In a virtual RFLP analysis with 17 restriction enzymes, the 16S rRNA sequence of the 'Ca. P. asteris' strain was found to match the pattern of the 16SrI-B subgroup. In addition, the phytoplasmas in the mixed-infection cases could be distinguished using specific primer sets. In conclusion, this study confirmed mixed infection of two phytoplasmas in one E. sylvestris plant, and also the presence of two phytoplasmas (of the 16SrⅠ and 16SrXXXⅡ groups) in Jeju Island (Republic of Korea).