Investigation of MIF gene promoter variations and their haplotypes in the Alzheimer disease in Turkish population: (Combined effect of two MIF gene promoter variations may play a role in Alzheimer’s disease)

In Alzheimer’s disease, which is characterized by amyloid plaques and neurofibrillary tangles in the brain tissue, many components such as acute phase proteins, cytokines, and proteases contribute to the progression of the disease or are part of the pathological process. The macrophage migration inhibitory factor (MIF) gene encodes a cytokine, which is secreted by lymphocytes, and has a role in the pathogenesis of autoimmune/inflammatory diseases such as rheumatoid arthritis. The purpose of this study to investigate the association between Alzheimer disease and MIF gene promoter polymorphisms. The 205 patients with Alzheimer disease (AD) and 130 age-sex matched healthy individuals were investigated in terms of MIF -173 G/C and MIF −794 CATT polymorphisms. The genotyping of MIF -173 G/C was determined using the RT-PCR method. MIF-794 CATT polymorphism was analyzed using PCR and DNA Sequencing. In terms of binary genotypes and haplotypes, the 5/5-GC (p = 0.004), 6/7-GG (p = 0.02) and, 6/6-GG (p = 0.026) binary genotypes, and 5-C (p = 0.003), 7-G (p = 0.026) and 6-G (p = 0.025) haplotypes were differed significantly between the patients and the controls. This is the first study investigating the relationship between AD and MIF in terms of different genotypes, haplotypes and, alleles. The fact that the binary genotype and allele distributions are significantly different between the patient and control group, suggests that this MIF variants may play a role in the pathogenesis of AD. © 202

Investigation of Brucella type bacteria in milk from Tokat province villages

Brucelloz, Brucella cinsi bakterilerin neden olduğu en sık görülen zoonotik hastalıklardan biridir. Hastalık sığır, koyun, keçi, köpek ve domuz gibi memeli hayvanların dişilerinde genital organlanna yerleşerek yavru atma, infertilite, mastitis, erken doğum, plasenta retensiyonu ve metritise; erkeklerde ise orşitise (erbezi iltihabı) neden olur. Bu çalışmada, Tokat ili ve ilçelerine ait köylerden toplanan sütlerde Brucelloz etmeninin prevalansının belirlenmesi amaçlanmıştır. Bu amaçla 161 inek, 58 koyun ve 33 keçi sütü örneklerinde Brucelloz etmeni Milk Ring Test (MRT) ve direkt ekim yöntemi ile araştırılmıştır. Çalışma sonucunda 161 inek sütünden 24'ü, 58 koyun sütünden 20'si ve 33 keçi sütünden 5'inin MRT sonucu pozitif bulunmuştur. Toplamda 252 adet çiğ süt örneğinden 49 adedinin, yani % 19.41'inin MRT sonucu pozitif bulunmuştur. MRT'si pozitif olan 49 örnekten yapılan kültür sonrası 2 adet süt örneğinden Brucella melitensis biyotip 3 izole edilebilmiştir. Brucella izole edilen 2 süt örneğinin de aynı sürüden 2 farklı koyuna ait olduğu belirlenmiştir. Böylece tüm çalışılan örneklerin % 0.79'unda, MRT pozitif çıkan örneklerin % 4.08'inde ve koyun sütü örneklerinin % 3.45'inde Brucella etkeni izole edilmiştir. Tokat ili ve ilçelerine ait köy ve kasabalardan toplanan sütlerle yapılmış ilk çalışma özelliğinde olan araştırmamız sonucunda örneklerin % 19.41 'inde MRT sonucu pozitif bulunmuş, % 0.79'unda ise etmen direkt ekim yöntemiyle izole edilmiştir. Böylece, bu çalışmayla Tokat ili ve ilçelerindeki hayvanlarda belirli oranda Brucella etmeninin var olduğu belirlenmiş ve süt ürünlerinin Brucelloz bakımından risk oluşturabileceğini göstermiştir.Brucellosis is one of the most common zoonotic diseases caused by Brucella. The disease is located genital organs in the females of mammals such as cattle, sheep, goats, dogs and pigs and cause to infertility, mastitis, preterm birth, placenta retention and metritis; in men it causes orchitis (gonorrhea inflammation). The bacteria can also spread from animals to humans and cause disease in humans. The aim of this study was to determine the prevalence of Brucellosis in milk collected from villages of Tokat province and districts. For this purpose, 161 cows, 58 sheep and 33 goat milk samples were investigated by Brucellosis agent with Milk Ring Test (MRT) and direct sowing method. At the end of the study, 24 of 161 cow's milk, 20 of 58 sheep's milk and 5 of 33 goat's milk were found to be positive by MRT. Of the 252 raw milk samples, 49 (n = 19.41%) were positive for MRT. Brucella melitensis biotype 3 were isolated from the 2 milk samples after the culturing from 49 samples with positive MRT. It was determined that 2 milk samples isolated from Brucea belong to 2 different sheep from the same herd. Thus, Brucella was isolated in 0.79% of all studied samples, 4.08% of MRT positive samples and 3.45% of sheep milk samples. As a result of our first study with milk collected from the villages and towns of Tokat province and districts, 19.41% of the samples were positive for MRT and 0.79% of the samples were isolated by direct planting method. Thus, in this study, it was determined that there is a certain amount of Brucella agent in Tokat province and its districts and it shows that dairy products may pose a risk for Brucellosis

The effects of endosulfan on the expression levels of DNA damage and apoptotic genes in HT22 cells: First preliminary study

*Rüstemoğlu, Aydın ( Aksaray, Yazar )Aim: The aim of this study was to determine the IC50 dose of endosulfan in the hippocampal HT22 cell line, and also to elucidate the effect of expression of DNA-PK, Bax, Bcl2 and Casp-3 genes involved in DNA repair and apoptotic pathway. Material and Methods: Cytotoxic effect of endosulfan and IC50 dose were determined by using XTT method after 24 hours of culture by applying endosulfan at 5 different concentrations (10, 25, 50, 75 and 100 μM) to HT22 cell lines. HT22 cells were then seeded into 6 sterile plates and treated with Endosulfan at IC50 for 12 hours. The expression changes of DNAPk, Bax, Bcl2 and Casp-3 genes, after total RNA isolation and cDNA formation, were determined by RT-PCR. Expression levels were calculated using the comparative 2-ΔΔCt method. Results: After 24 hours of endosulfan treatment at different doses in HT22 cell lines, a signifiant loss of viability was observed in all endosulfan treated groups. It was determined by XTT test, that the IC50 dose of endosulfan was 50 μM in 24 hours treatment. After 12 hours administration of IC50 endosulfan dose in HT22 cells following the examinations of DNA-PK and some apoptotic genes we observed different amounts of increases in expression as follows; 5-fold for DNA-PK, 18-fold for Bax, and 4-fold for Casp-3. On the other hand, approximately 2-fold decrease was detected in Bcl-2 gene. Conclusion: The IC50 dose of 24-hour endosulfan administration in HT22 cell lines was found to be 50 μM. Expression changes in the proapoptotic and antiapoptotic genes have shown that apoptosis is induced in endosulfan-administrated cells. In addition, the increase in DNA-PK gene expression suggests that endosulfan causes DNA damage in cells and triggers DNA repair mechanisms

Methylene-tetrahydrofolate reductase gene C677T and A1298C polymorphisms as a risk factor for Crimean-Congo hemorrhagic fever

Crimean-Congo hemorrhagic fever (CCHF) is a deadly viral disease. Methylene-tetrahydrofolate reductase (MTHFR) has an important role in folate metabolism, and also in the formation of new cells, DNA synthesis, repair and methylation. We aimed to examine the relationship between MTHFR gene C677T (Ala222Val, rs1801133) and A1298C (Glu429Ala, rs1801131) polymorphisms with CCHF in a Turkish population. Totally 273 participants were included in the current study. One hundred forty-one participants were CCHF patients and one hundred thirty-two participants were healthy controls. The polymerase chain reaction (PCR) and further restriction fragment length polymorphism (RFLP) assays were applied to determine the genotypes of MTHFR polymorphisms. We did not find any differences between the CCHF patients and healthy controls in terms of allele and genotype distributions of both the C677T and A1298C polymorphisms. After dividing the CCHF patients into different groups, we found that AC and AC + CC genotype frequencies of A1298C polymorphism were higher in Non-fatal patients compared to controls (p = 0.014 and p = 0.027, respectively). In composite genotype analysis between different groups, the frequency of CT-AA composite genotype, which is formed by C677T-A1298C polymorphisms, was found to be significantly higher in Mild CCHF patients compared to both Severe CCHF patients and controls (p = 0.036 and p = 0.008, respectively). In conclusion, in this study, we found a relationship between CCHF and MTHFR gene polymorphisms. Genotypes consisting C allele of A1298C polymorphism poses a risk for CCHF in Non-fatal group and CT-AA composite genotype of MTHFR gene C677T and A1298C polymorphisms showed a predisposition to Mild CCHF

Investigation of Associations between Obesity and LEP G2548A and LEPR 668A/G Polymorphisms in a Turkish Population

Objective. Obesity is a complex heterogeneous disease that is caused by genes, environmental factors, and the interaction between the two. The leptin (LEP) and leptin receptor (LEPR) genes have been evaluated for polymorphisms that could potentially be related to the pathophysiology of obesity and its complications. The aim of this study was to investigate the role of LEP G2548A and LEPR 668A/G polymorphisms in the pathogenesis of obesity. Subjects. The study included 127 patients with obesity and 105 healthy controls. Polymerase chain reaction and restriction fragment length analysis for LEP G2548A and LEPR 668A/G polymorphisms were applied. Results. There was no statistically significant difference in the genotype frequencies of the LEP gene polymorphism between patients and control groups (P>0.05). We found a difference in the LEPR genotypes between patients and controls, but this was not statistically significant (P=0.05). Additionally, we found an increased risk of obesity in the LEP/LEPR GG/GG combined genotype (P<0.05). Conclusion. Our findings indicate that the LEP G2548A polymorphism is not a relevant obesity marker and that the LEPR 668A/G polymorphism may be related to obesity in a Turkish population. Further researches with larger patient population are necessary to ascertain the implications of LEP and LEPR polymorphisms in obesity

Impact of endothelial NOS VNTR variant on susceptibility to diabetic neuropathy and type 2 diabetes mellitus

*Rüstemoğlu, Aydın ( Aksaray, Yazar )Purpose: The aim of this study was to evaluate whether the VNTR intron 4b/4a variant in the eNOS gene is associated with type 2 diabetes mellitus (T2DM) and DPN. Methods: A total of 598 subjects were enrolled in the study. eNOS VNTR 4b/4a variant was geno-typed by polymerase chain reaction (PCR) method. Results: eNOS VNTR intron 4b/4b genotype and b allele increased in patients with both DPN and T2DM compared healthy controls (p=0.0005, OR:1.94, p= 0.000002, OR:4.10, respectively). 4a/4b genotype was more prevalent in controls than in DPN and T2DM patients (p=0.00008, OR:0.46; p=0.000004, OR:0.24, respectively). eNOS VNTR b allele was more common in DPN patients and T2DM patients compared with controls (p=0.007, p=0.00002, respectively). Conclusion: The eNOS VNTR “4b/4b” homozygous genotype and hence “4b”allele as a genetic risk factor for T2DM and DPN, which may serve as a useful marker of increased susceptibility to the risk of these disorders

Evaluation of MIF -173 G/C Polymorphism in Turkish Patients with Ankylosing Spondylitis

Background: Ankylosing spondylitis (AS) is a chronic inflammatory disease mainly affecting the spine and sacroiliac joints. Macrophage migration inhibitory (MIF) factor is a regulatory cytokine that inhibits random immune cell migration. MIF gene promoter polymorphisms play a role in the progression of several inflammatory disorders. Aims: To investigate the relationship between the MIF gene -173 G/C single-nucleotide polymorphism (SNP) and AS. Study Design: Cross-sectional study. Methods: In this study, a total of 161 AS and 194 normal controls were recruited. The MIF gene -173 G/C SNP was analyzed by polymerase chain reaction using the restriction fragment length polymorphism method. Results: There was no significant difference between groups in terms of genotype distribution (p>0.05). When wild-type G/G and G/C+C/C genotypes are compared in terms of clinical characteristics, there is a significant difference between the average age and the duration of disease in AS patients (p<0.05). Conclusion: No significant relationship between AS disease and MIF -173 G/C polymorphism was found. MIF -173 G/C polymorphism (C allele) may affect the time of onset and the duration of disease in AS patients

Evaluation of MIF -173 G/C Polymorphism in Turkish Patients with Ankylosing Spondylitis

Background: Ankylosing spondylitis (AS) is a chronic inflammatory disease mainly affecting the spine and sacroiliac joints. Macrophage migration inhibitory (MIF) factor is a regulatory cytokine that inhibits random immune cell migration. MIF gene promoter polymorphisms play a role in the progression of several inflammatory disorders. Aims: To investigate the relationship between the MIF gene -173 G/C single-nucleotide polymorphism (SNP) and AS. Study Design: Cross-sectional study. Methods: In this study, a total of 161 AS and 194 normal controls were recruited. The MIF gene -173 G/C SNP was analyzed by polymerase chain reaction using the restriction fragment length polymorphism method. Results: There was no significant difference between groups in terms of genotype distribution (p>0.05). When wild-type G/G and G/C+C/C genotypes are compared in terms of clinical characteristics, there is a significant difference between the average age and the duration of disease in AS patients (p0.05). When wild-type G/G and G/C+C/C genotypes are compared in terms of clinical characteristics, there is a significant difference between the average age and the duration of disease in AS patients (p<0.05). Conclusion: No significant relationship between AS disease and MIF -173 G/C polymorphism was found. MIF -173 G/C polymorphism (C allele) may affect the time of onset and the duration of disease in AS patients