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Amaç: Bu çalışmada, hücre-hücre iletişiminin temel komponentleri olan eksozomların kronik, sistemik ve inflamatuvar bir hastalık olan ankilozan spondilit (AS) patogenezindeki olası rolü ve bu eksozomların kökeni araştırılmıştır.Gereç ve Yöntem: 10 AS hastası ve 10 sağlıklı kontrolden elde edilen serum içerisindeki eksozomlar, önce fiziksel özelliklerinden yararlanılarak Izon qEV2/35 nm kolonları ile, ardından biyokimyasal özelliklerinden yararlanılarak CD9, CD63 ve CD81 antikorlarıyla kaplı 3 µm çapındaki manyetik boncuklar ile izole edilmiştir. Filtratın saflığını değerlendirmek için BCA ve ELISA testleri yapılmıştır. Eksozom varlığını doğrulamak, eksozomların kökenini ve sitokin profilini ortaya çıkarmak için flow sitometri analizi ile eksozom yüzey belirteçleri karakterize edilmiştir. Eksozom konsantrasyonu ile ilgili flow sitometri bulgusunu desteklemek amacıyla, eksozom konsantrasyonu ile AS arasındaki ilişki BCA yöntemi ile test edilmiş ve sonuçlar karşılaştırılmıştır.Bulgular: Flow sitometri analizinde, AS grubunun CD86+TSG101+ ve CD3+TSG101+ eksozom yüzdeleri kontrol grubuna göre anlamlı derecede yüksekti (p

Objective: In the present study, it was aimed to study the antioxidant effects of spironolactone (SPL) to determine its possible protective effects in hepatic ischemia reperfusion injury. Material and Methods: Hepatic artery, portal vein, and bile duct of Wistar albino rats were clamped for 45 minutes under anesthesia to form an ischemia period. Then reperfusion was allowed and the rats were decapitated 60 minutes later. SPL (20 mg/kg, p.o.) or SF was orally administered for 30 minutes before ischemia. Rats in the control arm underwent sham surgery and were administered isotonic saline. Liver function was studied by measuring aspartate aminotransferase (AST), alanine aminotransferase (ALT), tumor necrosis factor-alpha (TNF-alpha), and interleukin 1beta (IL-1 beta) levels. Malondialdehyde (MDA), glutathione (GSH), luminol, and lucigenin levels, myeloperoxidase (MPO) and Na+-K+- ATPase enzyme activities were analyzed to study tissue injury under light microscope. Results: While IR increased AST, ALT, TNF-alpha, and IL-1 beta levels and MDA, luminol, and lusigenin levels and MPO activities, it caused a decrease in GSH levels and Na+K+-ATPase activity. Spironolactone administration significantly improved these values. Conclusion: Protective effects of SPL against ischemia/reperfusion injury via various mechanisms suggest that this agent may become a novel treatment agent in clinical practice.

WOS:000215712700012 Purpose: The present study was conducted to characterize the possible therapeutic effects of glucagon-like peptide (GLP)-1 and GLP-2 against oxidative damage in the ileum and colon of irradiated rats. Methods and materials: Sprague-Dawley rats of both sexes received either a single dose of GLP-1 (0.1 nmol/kg, intraperitoneally, ip; n = 6) 10 min before abdominal irradiation (IR) or two consecutive doses of GLP-2 (7 nmol/kg, ip; n = 6) at 30 and 10 min before IR, while another group was administered vehicle (n = 6) 10 min before IR. Control rats (n = 6) received vehicle treatment without IR. On the fourth day of IR, samples from ileum and colon were removed for histological analysis, for the determination of myeloperoxidase (MPO) activity, malondialdehyde (MDA) and glutathione (GSH) levels, as well as DNA fragmentation ratio, an index of apoptosis. Results: IR-induced oxidative injury in the colonic tissue of vehicle-treated rats, evidenced by elevated MDA levels and MPO activity, as well as depleted colonic GSH levels, was reversed by GLP-2, while GLP-1 reduced IR-induced elevations in colonic MDA levels. IR-induced injury with elevated ileal MDA levels was reduced by GLP-1, while replenishment in GSH was observed in GLP-2-treated rats. Conclusion: Current findings suggest that GLP-1 and GLP-2 appear to have protective roles in the irradiation-induced oxidative damage of the gut by inhibiting neutrophil infiltration and subsequent activation of inflammatory mediators that induce lipid peroxidation. Copyright (C) 2015, The Egyptian Society of Radiation Sciences and Applications. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license.

Obstructive jaundice (OJ) can be defined as cessation of bile flow into the small intestine due to benign or malignant changes. Nesfatin-1, recently discovered anorexigenic peptide derived from nucleobindin-2 in hypothalamic nuclei, was shown to have anti-inflammatory and antiapoptotic effects. This study is aimed to investigate the therapeutic effects of nesfatin-1 on OJ in rats. Twenty-four adult male Wistar-Hannover rats were randomly assigned to three groups: sham (n = 8), control (n = 8), and nesfatin (n = 8). After bile duct ligation, the study groups were treated with saline or nesfatin-1, for 10 days. Afterward, blood and liver tissue samples were obtained for biochemical analyses, measurement of cytokines, determination of the oxidative DNA damage, DNA fragmentation, and histopathologic analyses. Alanine aminotransferase and gamma-glutamyl transferase levels were decreased after the nesfatin treatment; however, these drops were statistically non-significant compared to control group (p = 0.345, p = 0.114). Malondialdehyde levels decreased significantly in nesfatin group compared to control group (p = 0.032). Decreases in interleukin-6 and tumor necrosis factor-α levels from the liver tissue samples were not statistically significant in nesfatin group compared to control group. The level of oxidative DNA damage was lower in nesfatin group, however this result was not statistically significant (p = 0.75). DNA fragmentation results of all groups were similar. Histopathological examination revealed that there was less neutrophil infiltration, edema, bile duct proliferation, hepatocyte necrosis, basement membrane damage, and parenchymal necrosis in nesfatin compared to control group. The nesfatin-1 treatment could alleviate cholestatic liver damage caused by OJ due to its anti-inflammatory and antioxidant effects.

1.ÖZETSpondiloartropati (SpA) hastalık grubunda yer alan Ankilozan spondilit (AS); başlıca enflamatuvar sırt ağrısı, asimetrik periferal oligoartrit, entezit ile tanımlanmış, diğer çok etkenli hastalıklar gibi, ortaya çıkışı genetik ve çevresel etkenlerle tetiklenen romatizmal otoimmün bir hastalıktır. Geçtiğimiz yıllar içinde insan ve diğer organizma genomlarının dizilenmesiyle varyasyonların incelenmesi, birçok hastalıkta olduğu gibi, AS hastalığının da genetik temelinin anlaşılması konusunda anlamlı mesafe kaydedilmesine imkan sağlamıştır.Genetik temelin yanı sıra epigenetik değişikliklerin de fizyolojik süreçlerde olduğu kadar, hastalık gelişiminde de önemli olduğu bilinmektedir. Epigenetik değişiklikler, DNA dizisindeki değişimlerle açıklanamayan gen ifadesindeki kalıtsal değişiklikler olarak tanımlanır. Başlıca epigenetik değişiklikler; DNA metillenmesi, histon ve kromatin yapı değişiklikleri ve kodlamayan RNA havuzundan oluşur. Yapılan çalışmalarda, epigenetik özellikleri etkileyen moleküllerin ifade edilmesinde ortaya çıkan değişikliklerin artirit gelişimi ve romatoid artiritin patogeneziyle ilişkisi olduğu rapor edilmiştir.Bu çalışmada AS tanısı almış diskordant monozigotik ikizler ile hastalığın radyolojik skorlamasına göre ağır ve hafif grup olarak klinikte tanımlanan iki gruba dahil edilmiş hastalarda epigenetik yapıyı düzenleyen (DNA metilaz, histon metilaz, kromatin şekillendiriciler gibi) genlerin ekspresyonları analiz edilmiştir. Analiz sonucunda NEK6, PRMT6, KDM6B, AURKA, CARM1, SETD1B, SETDB1, PRMT1, DNMT1, PRMT3, SETD7, MBD3, HDAC1, KDM4A, MLL, SUV420H1, NCOA3, KAT2B, SMYD3, RPS6KA5 genlerinin ekspresyonlarında görünen gruplar arası farklılık, anılan genlerin daha yüksek sayıda hasta gruplarında ekspresyon açısıdan incelenerek, doğrulanması gerektiğini ortaya koymuştur. Anahtar Kelimeler: Spondiloartropati , Ankilozan Spondilit, Epigenetik, Epigenetik Modifiye Edici1.SUMMARYAnkylosing spondylitis (AS), which is a member of multifactorial autoinflammatory Spondyloarthropathies, is defined by inflammatory dorsal pain, asymmetric peripheral oligoarthritis, enthesitis.The analysis of variations by DNA sequencing in human and other organisms has enabled a significant progress in understanding of the genetic basis of AS and many other diseases. Beside genetical basic, epigenetic changes are also considered as important tools for physiological processes and disease development. Major epigenetic changes can be classified as DNA methylation, histone modifications, chromatin remodelling and non-coding RNA effects. Studies so far are indicating that expression changes of molecules involved in epigenetic modifications might have a role in development of arthritis and pathogenesis of rheumatoid arthritis.In this study the expression of so called “epigenetic modifiers” like DNA methylase, histon methylase, chromatin remodeller complex has been analysed in a discordant monozygotic twin couple for AS and AS patients that have been classified as “fast progressing” and slow progressing according to radiographic scoring. The differential expression in NEK6, PRMT6, KDM6B, AURKA, CARM1, SETD1B, SETDB1, PRMT1, DNMT1, PRMT3, SETD7, MBD3, HDAC1, KDM4A, MLL, SUV420H1, NCOA3, KAT2B, SMYD3, RPS6KA5 genes should be validated in the same patient groups with larger number of subjects. Key Words: Spondyloarthropathy, Ankylosing Spondylitis, Epigenetic, Epigenetic Modifier

Background: Oxidative stress is a mediator of secondary injury to the spinal cord following trauma. Objective: To investigate the putative neuroprotective effect of a-lipoic acid (LA), a powerful antioxidant, in a rat model of spinal cord injury (SCI). Methods: Wistar albino rats were divided as control, vehicle-treated SCI, and LA-treated SCI groups. To induce SCI, a standard weight-drop method that induced a moderately severe injury (100 g/cm force) at T10 was used. Injured animals were given either 50 mg/kg LA or saline at 30 minutes postinjury by intraperitoneal injection. At 7 days postinjury, neurologic examination was performed, and rats were decapitated. Spinal cord samples were taken for histologic examination or determination of malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity, and DNA fragmentation. Formation of reactive oxygen species in spinal cord tissue samples was monitored by using a chemiluminescence (CL) technique. Results: SCI caused a significant decrease in spinal cord GSH content, which was accompanied with significant increases in luminol CL and MDA levels, MPO activity, and DNA damage. Furthermore, LA treatment reversed all these biochemical parameters as well as SO-induced histopathologic alterations. Conversely, impairment of the neurologic function caused by SCI remained unchanged. Conclusion: The present study suggests that LA reduces SCI-induced oxidative stress and exerts neuroprotection by inhibiting lipid peroxidation, glutathione depletion, and DNA fragmentation.

1.ÖZET: GADD45 gen ailesi hücrede DNA hasarına cevap oluşturulması ve hücre bölünmesinin durdurulması süreçlerinde etkili olan, nükleer proteinler GADD45α,β ve γ’ yı kodlar. Her üç GADD45 gen ailesi üyesi de MTK1 kinazı aktive ederek p38/c-jun-NH2-kinaz aktivasyonunu ve apoptozu indüklemektedir. Yapılan çalışmalarda GADD45γ’nın tümör hücrelerinin koloni oluşturmasını ve bölünmesini baskıladığı gösterilmiştir. Ayrıca birçok kanser türünde GADD45γ’nın metile durumda bulunduğu ve metilasyonun ekspresyonu baskıladığı gösterilmiştir. Yapılan çalışmanın amacı hematolojik malignitelerde GADD45γ promotorunun metilasyon durumunun belirlenmesi ve metilasyonun ekspresyona etkisinin saptanmasıdır. Bunun için hemotolojik malignite hücre soylarından ve 117 akut miyeloid lösemi (AML), 94 multipl miyelom (MM) teşhisi konmuş hastadan alınan kan ve kemik iliği örneklerinden DNA izolasyonu yapılmıştır. Bisülfit modifikasyonunun ardından örneklerle metilasyon spesifik polimeraz zincir reaksiyonu (MSP) yapılmıştır. Non-Hodgkin lenfoma hücre serileri L–428 ve L–1236, Burkitt lenfoma hücre serisi Raji hücrelerinde GADD45γ promotoru tamamiyle metillenmiş olduğu, AML hücre serileri GDM–1, HL–60, KG1a ve MM hücre serisi OPM–2 hücrelerinde ise kısmen metillenmiş olduğu görülmüştür. İncelenen AML hasta örneklerinde metilasyon saptanmazken 94 MM hasta örneğinden 8’inde metilasyon saptanmıştır. İstatistiksel hesaplamalar sonucunda metilasyon saptanan hastalarda ortalama kalsiyum düzeyinin metilasyon görülmeyenlere oranla daha yüksek olduğu görülmüştür. Ayrıca GADD45γ’nın metile durumda olduğu hastaların ortalama sağ kalım sürelerinin 13,7 ay, genin metillenmemiş olarak bulunduğu hastalarda ise bu sürenin 57,1 ay olduğu görülmüştür. Metilasyonun genin ekspresyonuna etkisinin belirlenmesi amacıyla hücre soyları DNA metiltransferaz (DNMT) inhibitörü DAC ile 96 saat boyunca inkübe edilmiş ve RT-PCR uygulanmıştır. GADD45γ promotorunun tamamiyle metillenmiş durumda bulunduğu L–1236 hücrelerinde DAC uygulaması sonrası genin ekspresyonunda değişiklik olmazken, Raji, L–428, HL–60, KG1a, OPM–2 hücrelerinde DAC uygulaması sonrası genin ekspresyonunda artış gözlemlenmiştir. Elde edilen bulgular hemotolojik malignitelerde GADD45γ geninin tümör tipine bağlı olarak epigenetik mekanizmalarla sessizleştirilebileceğini göstermiştir.Anahtar Kelimeler: GADD45 gen ailesi, GADD45 gamma, epigenetik, DNA metilasyonu, hematolojik malinite 1.SUMMARYEpigenetic dysregulation of GADD45 in hematologic malignanciesGADD45 gene family encodes the nuclear proteins, GADD45α, β and γ, which response to damage in DNA in a cell and stopping cell proliferation. It has been shown from the research conducted in recent years that GADD45γ suppresses the colony formation and proliferation of tumor cells.The purpose of this study is to determine the methylation state of GADD45γ promoter in hemotologic malignities and the effect of methylation on expression. Therefore, DNA isolation is performed on blood and bone marrow samples of 117 acute myeloid leukemia (AML), and 94 multiple myelom (MM) patients and hematologic malignency cell lines . After the bisulfide modification of DNA, methylation specific polymerase chain reaction (MSP) has been performed on the samples. It has been found that, in Hodgkin lymphoma cell lines L-428 and L-1236 and Burkitt lymphoma cell lines Raji, GADD45γ promoter was fully methylated. However in AML cell lines GDM-1, HL-60, KG1a and MM cell lines OPM-2 , GADD45γ promoter has been found to be partially methylated. No methylation was found in AML patient samples whereas 8 of of 94 MM patient samples show promoter methylation. The survival time of the patients with methylated GADD45γ is 13.7 months, whereas the survival time of the patients with unmethylated GADD45γ is 57.1 months. In order to understand the effect of methylation on the gene expression, the cell lines were treated with DNA methyltransferase inhibitor (DAC) for 96 hours and then real-time polymerase-chain reaction (RT-PCR) was performed. In L-1236 cells where GADD45γ promoter was fully methylated, there has been no change in gene expression after DAC treatment whereas in Raji, L-428, HL-60, KG1a and OPM-2 cells, an increase in gene expression has been observed after DAC treatment,. The findings of this study indicate that in hemotoligic malignancies, GADD45γ gene may be silenced in epigenetic mechanisms according to the type of tumor.Key Words: GADD45 gene family, GADD45 gamma, Epigenetic, DNA methylation, heamatological malignancy