JAK2 mutasyonlarının Kronik Miyeloproliferatif hastalık ayırıcı tanısındaki önemi : The importance of JAK2 mutations in distinctive diagnosis of chronic myeloproliferative disorders

1. Miyeloproliferatif hastalıklar, mutant multipotent hematopoetik kök hücre kaynaklı olan birçok kronik hematolojik hastalığı bünyesinde barındırmaktadır. Son çalışmalarda, JAK2 geninde oluşan V617F somatik mutasyonu, polisitemia vera, esansiyel trombositoz ve idiyopatik miyelofibrozda rapor edilmiştir. JAK2 bir sitoplazmik tirozin kinazı kodlamaktadır. JAK homoloji 2 (JH2) negative regülator bölgesinde meydana gelen V617F mutasyonunun, JAK2 kinaz etkinliğini arttırdığı ve sitokinden bağımsız büyümeye neden olduğu hücre serileri ve kültür ortamındaki kemik iliği hücrelerinde gösterilmiştir. Polisitemia vera (PV), esansiyel trombositoz (ET) ve miyeloid metaplazili miyelofibroz (MMM) hastalıklarında keşfedilen JAK2V617F mutasyonları bu hastalıkları ileri düzeyde anlamak adına çok önemli bir gelişme olarak kabul edilmiştir. Çoğu PV ile birçok ET ve MMM hastaları JAK2V617F mutasyonu açısından pozitif olup, bu mutasyon hematopoetik kök hücrelerde meydana gelmektedir. JAK2 aktif bir tirozin kinaz olup, JAK/STAT yolağını eritropoetin reseptörü (EPOR), trombopoetin reseptörü veya granülosit koloni-uyarıcı faktör reseptörü (GCSFR) ile birlikte daha etkin aktifleştirebilme özelliğine sahiptir. Kemik iliği transplantasyonu ile JAK2V617F mutasyonu oluşturulmuş fare çalışmalarında, mutasyonun PV veya miyelofibrozu tetiklediği gösterilmiş, kronik miyeloproliferatif hastalık patogenezinde merkezi bir rol üstlendiği desteklenmiştir. Bu çalışmada, JAK2V617F mutasyon varlığının kronik miyeloproliferatif hastalıkların ayırıcı tanısındaki öneminin gösterilmesi amaçlanmıştır. Mutasyonun somatik olması, hassasiyetin önem kazanmasını sağlamış, bu yüzden çalışmada hibridizasyon prob yöntemini takip eden melting curve analizi tercih edilmiştir. Elde edilen bulgular, mutasyonun dağılımı, mutasyonunun etkileri ve endikasyon kriterlerinin önemine işaret etmiş, yaş, cinsiyet, yeterli hastalık hikayesi gibi etkenler de kullanılarak daha kapsamlı bir çalışma yapılmasını zorunlu kılmıştır. Anahtar Sözcükler Esansiyel Trombositoz, JAK2V617F mutasyonu, Kronik miyeloproliferatif hastalık, Miyeloid metaplazili miyelofibroz, Polisitemia Vera 2. SUMMARY THE IMPORTANCE OF JAK2V617F MUTATION IN DISTINCTIVE DIAGNOSIS OF CHRONIC MYELOPROLIFERATIVE DISORDERS The myeloproliferative disorders comprise a spectrum of chronic hematologic diseases that are likely to arise from a mutant multipotent hematopoietic stem cell. In recent researches, V617F somatic mutation in the Janus kinase 2 gene (JAK2) has been detected in polycythemia vera, essen¬tial thrombocythemia or idiopathic myelofibrosis patients. This gene encodes a cytoplasmic tyrosine kinase. V617F mutation, which occurs in the JAK homology 2 (JH2) negative regulatory domain, is shown to increase the JAK2 kinase activity thus causing cytokine-independent growth in cell lines and cul¬tured bone marrow cells. The identification of JAK2V617F mutation in polycythemia vera (PV), essential thrombocytosis (ET), and myelofibrosis (MF) represents an important advance in understanding these myeloproliferative disorders (MPD). Most patients with PV and a significant number of patients with ET or MF are JAK2V617F positive, the mutation likely to be arising in the hematopoietic stem cell lineage. JAK2 is a constitutively active tyrosine kinase that is able to activate JAK / STAT signaling most efficiently when co-expressed with erythropoietin receptor (EPOR), the thrombopoietin receptor (MPL), or the granulocyte colony-stimulating factor receptor (GCSFR). Data from murine models supports the central role of JAK2V617F in the pathogenesis of MPD, as expression of JAK2V617F in a bone marrow transplantation assay results in polycythemia and myelofibrosis in recipient mice. This study is intended to show the importance of the presence of JAK2V617F mutation in distinctive diagnosis of chronic myeloproliferative disorders. Because the mutation is somatic, the sensitivity has become more significant and melting curve analysis following the hybridization prob method has been preferred. The resulting findings has pointed to the importance of mutation distribution, mutation effects and indication criteria and it has made it necessary to do a more comprehensive study using the factors such as the age, gender and sufficient disorder story. Key Words Essential thrombocytosis, JAK2V617F mutation, Chronic myeloproliferative disorders, Myelofibrosis with myeloid metaplasia, Polycythemia ver

Association ofNFKB1A and MicroRNAs Variations and the susceptibility to Atherosclerosis

ds

Pleiotropic effects of pitavastatin: A pilot study using the saphenous vein endothelial cell model of endothelial injury and prevention of atherosclerosis

OBJECTIVE: Cardiovascular diseases are responsible for the majority of deaths on a global scale. Atherosclerosis is the main risk factor for cardiovascular disorders and represents a complex phenomenon associated with endothelial dysfunction and inflammation. Statins, especially atorvastatin (ATV) and pitavastatin (PTV), are common agents used to control ongoing atherosclerotic events in the body to minimize cardiovascular disease-based deaths. MATERIALS AND METHODS: The present study aimed at comparing the efficacy of ATV and PTV in a cell line model of inflammation. Human saphenous vein cells were treated with TNF-alpha to mimic atherosclerotic conditions, and the cells were divided into 7 groups, including control, DMSO, TNF-alpha (10 ng/mL-6 hours), ATV (50 μM/24 hours), PTV (2 μM/24 hours), ATV (50 μM/24 hours)+TNF-alpha (10 ng/ mL-6 hours) and PTV (2 μM/24 hours)+TNF-alpha (10 ng/mL-6 hours). The expression levels of 20 proinflammatory cytokines and chemokines were investigated in these groups using a human atherosclerosis antibody array. RESULTS: Possible pathway interactions were determined by STRING and PANTHER analyses. Comparison with the effect of ATV indicated that PTV reduced the levels of 4 proinflammatory cytokines: CCL11, CSF2, CCL20, and TGFB1 (p<0.05). CONCLUSIONS: Pleiotropic effects of pitavastatin against cardiovascular diseases appeared to be better; however, additional studies are required to compare statins and to identify new drugs that maintain broader protection from the risks of cardiovascular diseases

The relationship between elevated plasma zonulin levels and Hashimoto's thyroiditis

Background/aim: Hashimoto thyroiditis (HT) is one of the most prevalent autoimmune diseases. The intestine microbiota is strongly associated with autoimmune diseases. Zonulin, a modulator of tight junctions that controls the selective permeability of the intestine can induce an elevation in gut permeability. We aimed to investigate the association of plasma zonulin levels with HT. Materials and methods: We compared 77 HT patients with 66 age-gender and BMI-matched healthy individuals in the case of plasma zonulin levels. Plasma zonulin levels were measured by ELISA. The statistical analyses were performed using Student's t-test and chi-square tests. The predictive power was investigated using univariate and multivariate logistic regression analysis. Results: We found that the increase in plasma zonulin levels in the HT group was statistically significant compared to the control group (p < 0.001). The regression analysis showed that urea, anti-thyroid peroxidase, aspartate aminotransferase, thyroid-stimulating hormone, free T3, and serum zonulin levels were found to be associated with HT in both univariate and multivariate models (p < 0.05). Conclusion: Zonulin is a possible biomarker candidate that may link intestinal permeability with the etiology of autoimmune diseases

NFKB1 rs28362491 and pre-miRNA-146a rs2910164 SNPs on E-Cadherin expression in case of idiopathic oligospermia: A case-control study

Background: A notable proportion of idiopathic male infertility cases is accompanied by oligozoospermia; and yet, the molecular mechanisms of fertilization problem underlying this defect are still unclear. Epithelial cadherin has been involved in several calcium-dependent cell-to-cell adhesion events; however, its participation in gamete interaction has also not been fully investigated. Objective: The aim was to investigate the changes in the expression of E-cadherin, based on the frequency of Single nucleotide polymorphisms in Nuclear Factor Kappa-B 1 and pre-mir-146a in oligospermic men. Materials and Methods: In this case-control study, semen and blood samples of 131 oligospermic men as the case group and 239 fertile healthy men as the control group were analyzed. Variants single nucleotide polymorphisms rs28362491 and rs2910164 were performed using polymerase chain reaction-restriction fragment length polymorphism method and E-cadherin expression were determined by immunoprecipitation studies. Results: ins/ins genotype of rs28362491 was determined as a risk factor for idiopathic oligospermia by 1.73 times (p=0.0218), whereas no significant differences were found between the groups concerning pre-mir-146a rs2910164 polymorphism (p=0.2274 in case of GC genotype and p=0.9052 in case of GG genotype). Combined genotype analysis results did not show any notable differences between the multiple comparisons of 28362491-rs2910164 in oligospermic men and control groups. In addition, E-cadherin expression of oligospermic men with ins/ins genotype was significantly lower than patients with del/ins genotype (p=0.0221). E-cadherin expression level was low in oligospermic men with respect to the control group in presence of ins/ins genotype of NFKB1 gene. Conclusion: These results suggest that ins allele prevents binding of surface proteins to spermatozoa, leading to a low affinity of sperm-oocyte interaction in oligospermic men