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Öğe Advances in cryopreservation of bull sperm(FRONTIERS MEDIA SA, 2019) Uğur, Muhammet Raşit; Abdelrahman, Amal Saber; Evans, Holly C.; Gilmore, Alicia A.; Hitit, Mustafa; Arifiantini, Raden Iis; Purwantara, Bambang; Kaya, Abdullah; Memili, ErdoğanCryopreservation of semen and artificial insemination have an important, positive impact on cattle production, and product quality. Through the use of cryopreserved semen and artificial insemination, sperm from the best breeding bulls can be used to inseminate thousands of cows around the world. Although cryopreservation of bull sperm has advanced beyond that of other species, there are still major gaps in the knowledge and technology bases. Post-thaw viability of sperm is still low and differs significantly among the breeding bulls. These weaknesses are important because they are preventing advances both in fundamental science of mammalian gametes and reproductive biotechnology. Various extenders have been developed and supplemented with chemicals to reduce cryodamage or oxidative stress with varying levels of success. More detailed insights on sperm morphology and function have been uncovered through application of advanced tools in modern molecular and cell biology. This article provides a concise review of progress in the cryopreservation of bull sperm, advances in extender development, and frontiers using diverse techniques of the study of sperm viability. This scientific resource is important in animal biotechnology because with the advances in discovery of sperm fertility markers, there is an urgent need to improve post-thaw viability and fertility of sperm through enhanced cryopreservation for precision agriculture to produce food animals to ensure food security on the global scale.Öğe Alcoholic extract of Tarantula cubensis induces apoptosis in MCF-7 cell line.(ALLIED ACAD, 2017) Er, Ayse; Corum, Orhan; Corum, Duygu; Hitit, Mustafa; Donmez, Huseyin; Guzeloglu, AydinTarantula cubensis Alcoholic Extract (TCAE) is a homeopathic agent used for treating many disorders. This study aimed to define the effects of TCAE on the breast carcinoma cell line (MCF-7). After various concentrations (10, 20, 40, 80 and 160 mu l/ml) of TCAE were applied to MCF-7 cells and the human embryonic kidney cell line (HEK293), the cells were incubated for 1, 3, 6, 9, 12, 24 and 48 h, followed by analysis by MTT assays. According to the results of the MTT assays, cells treated with 20 or 40 mu l/ml TCAE for 6 h were applied to apoptosis analysis by flow cytometry. Secreted levels of tumor necrosis factor alpha (TNF alpha), interleukin (IL)-1 beta, IL-6, IL-10, Interferon-gamma (IFN gamma), Transforming Growth Factor beta (TGF beta), and Nuclear Factor-kappa B (NF-kappa B) were measured using ELISAs. TNF alpha and TGF beta levels increased while IL-6 and IL-10 levels fluctuated in MCF-7 cells. In conclusion, our study suggests that TCAE may change the normal cancer physiology and lead to cell death by activating apoptosis in MCF-7 cells.Öğe Bir keçide hermafroditizm olgusu: Klinik ve genetik yaklaşım(2014) Özel, Çağlayan; Şen, Gonca; Hitit, Mustafa; Koçak, Nadir; Güzeloğlu, Aydın; Semacan, Ahmet; Kurar, ErcanBu vaka takdiminde, 2010 yılında Selçuk Üniversitesi, Veteriner Fakültesi, Doğum ve Jinekoloji Kliniğine getirilen ve hermafroditizm olgusu saptanan Maltız melezi bir keçinin klinik muayenesi ile sitogenetik ve moleküler genetik analizleri gerçekleştirildi. Anamnezde keçinin tek olarak doğduğu, klinik muayenede boynuzsuz, klitorisin 1.9x0.9 cm büyüklüğünde, vajinanın normalden daha kısa olduğu gözlendi. Palpasyon ve ultrasonografik muayenede inguinal bölgede, deri altında sağ (4.88 cm) ve sol (2.87 cm) testis tespit edildi. Meme ve meme başları normal konumunda ve büyüklükte belirlendi. Deri altı rüdimenter penis varlığı tespit edildi, ancak scrotum ve açık prepisyum gözlenmedi. Karyotip analizi için periferik kandan hazırlanan kromozom preparatları GTG bantlama tekniği ile boyanmış ve image analyzer programı ile uygun metafazlar belirlenerek analiz edildi. Kan örneğinden DNA izolasyonu geçekleştirildi, sex-determining region Y (SRY) ve amelogenin gen bölgeleri pozitif ve negatif kontroller kullanılarak polimeraz zincir reaksiyonu (PZR) ile yükseltgendi. Karyotip analizinde metafaz plağı 60,XY olarak tespit edildi. PZR analizlerinde SRY lokusu negatif, amelogenin lokusunda ise yalnızca AMX alleli tespit edildi. Keçilerde yaygın olarak gözlenen hermafroditizm vakalarının şekillenmesinde farklı mekanizmalar bildirilmektedir. Hermafroditizm vakalarının karyotip ve moleküler genetik analizler ile detaylı incelenmesi ve boynuzsuz keçilerin damızlıktan çıkarılması tavsiye edilmektedir.Öğe Bir keçide hermafroditizm olgusu: Klinik ve genetik yaklaşım(Selçuk Üniversitesi Veterinerlik Fakültesi, 2014) Özel, Çağlayan; Şen, Gonca; Hitit, Mustafa; Koçak, Nadir; Güzeloğlu, Aydın; Semacan, Ahmet; Kurar, ErcanBu vaka takdiminde, 2010 yılında Selçuk Üniversitesi, Veteriner Fakültesi, Doğum ve Jinekoloji Kliniğine getirilen ve hermafroditizm olgusu saptanan Maltız melezi bir keçinin klinik muayenesi ile sitogenetik ve moleküler genetik analizleri ger- çekleştirildi. Anamnezde keçinin tek olarak doğduğu, klinik muayenede boynuzsuz, klitorisin 1.9x0.9 cm büyüklüğünde, vajinanın normalden daha kısa olduğu gözlendi. Palpasyon ve ultrasonografik muayenede inguinal bölgede, deri altında sağ (4.88 cm) ve sol (2.87 cm) testis tespit edildi. Meme ve meme başları normal konumunda ve büyüklükte belirlendi. Deri altı rüdimenter penis varlığı tespit edildi, ancak scrotum ve açık prepisyum gözlenmedi. Karyotip analizi için periferik kandan hazırlanan kromozom preparatları GTG bantlama tekniği ile boyanmış ve image analyzer programı ile uygun metafazlar belirlenerek analiz edildi. Kan örneğinden DNA izolasyonu geçekleştirildi, sex-determining region Y (SRY) ve amelogenin gen bölgeleri pozitif ve negatif kontroller kullanılarak polimeraz zincir reaksiyonu (PZR) ile yükseltgendi. Karyotip analizinde metafaz plağı 60,XY olarak tespit edildi. PZR analizlerinde SRY lokusu negatif, amelogenin lokusunda ise yalnızca AMX alleli tespit edildi. Keçilerde yaygın olarak gözlenen hermafroditizm vakalarının şekillenmesinde farklı mekanizmalar bildirilmektedir. Hermafroditizm vakalarının karyotip ve moleküler genetik analizler ile detaylı incelenmesi ve boynuzsuz keçilerin damızlıktan çıkarılması tavsiye edilmektedir.Öğe A Cardioprotective Role of Nerium oleander with the Expression of Hypoxia Inducible Factor 2A mRNA by Increasing Antioxidant Enzymes in Rat Heart Tissue(UNIV FED RIO GRANDE DO SUL, 2018) Hitit, Mustafa; Corum, Orhan; Corum, Duygu Durna; Donmez, Huseyin; Cetin, Gul; Dik, Burak; Er, AyseBackground: Nerium oleander (NO) distillate is used to either protect heart cells against oxidative stress or reduce the risk of cardiovascular disease by regulating the production of reactive oxygen species. Hypoxia-inducible factors (HIFs) regulate cellular antioxidant defense mechanisms under hypoxic conditions in which heart cells survive; however, the key responsible mechanism of NO distillate for cardioprotection remains elusive. The objective of this study was to evaluate the effects on heart tissue at different time intervals after administering NO distillate intraperitoneally (IP) while considering the transcriptional regulation of HIFs and representative antioxidant enzymes. Materials, Methods & Results: The NO plant was chopped, and distillated water was added. The mixture was distilled, and the distillate separated and collected into tubes, after which it was lyophilized to obtain dry material. Twenty male Wistar albino rats (2-3 month-old, 250-300 g each) were used in the study. The rats were randomly divided into four groups. The control group (n = 5) received IP injections of saline; the remaining 15 rats received IP injections of a single dose of 7.5 mL NO distillate. The NO distillate injected rats were divided into three groups according to the time from injection to harvest the heart tissue samples. The tissues were collected at 0 h (control; n = 5), 2 h (group 2; n = 5), 4 h (group 3; n = 5), and 8 h (group 4; n = 5) after injection and under general anesthesia (60 mg/kg ketamine, IP + 10 mg/kg xylazine, IP). Quantitative polymerase chain reaction (qPCR) was used to assess the expression profiles of the genes of interest in the heart tissues. Hypoxanthine phosphoribosyltransferase was used as the reference gene. The expression of manganese superoxide dismutase (MnSOD) mRNA was in a steady state level between the control group and group 2 (P > 0.05); however, it significantly increased in group 3 and 4 compared with that in the control (P < 0.05). Expression of catalase (CAT) mRNA was significantly higher in group 2 than in the control group (P < 0.05) although it was lower in group 3 and 4 than in group 2 (P < 0.05); however, it appeared to be similar among the control group, group 3, and group 4 (P > 0.05). Copper (Cu) SOD mRNA was equally expressed in both the control group and group 2 (P > 0.05) but was lower in group 3 and 4 than in group 2 (P < 0.05). Expressions of HIF1A, HIF2A, and HIF3A mRNA were detected in the rat heart tissues in the control and 2, 4, and 8 h after administration of NO distillate. Expression of HIF1A mRNA was in a steady state and did not differ among groups 2, 3, and 4 (P > 0.05). Similarly, the expression of HIF2A mRNA did not change between the control group and group 2 (P > 0.05); however, it was higher in group 3 than in the control (P < 0.05) and tended to be higher in group 3 than in group 2 (P = 0.063). HIF3A mRNA expression did not change significantly in the heart tissue of any of the groups (P > 0.05). Discussion: The present study using rats determined that MnSOD, CAT, CuSOD, HIF1A, HIF2A, and HIF3A mRNA are expressed in the heart tissues after administration of NO distillate. The increased expression of HIF2A mRNA after 4 h in accordance with a rise in CAT mRNA after 2 h, and MnSOD mRNA after 4 and 8 h might confirm the role of HIF2A mRNA in oxidative stress defense by regulating antioxidant enzymes; consequently, this study may expand our understanding of uses of NO distillate with respect to molecular pathways.Öğe Comparison of reverse-transcriptase polymerase chain reaction (RT-PCR) and rapid test for the detection of bovine rotavirus and bovine coronavirus in anatolian water buffaloes(Selçuk Üniversitesi Veteriner Fakültesi, 2020) Bulut, Oya; Uyunmaz Saklı, Gülşah; Hasöksüz, Mustafa; Dik, Irmak; Hadimli, Hasan Hüseyin; Hitit, MustafaAmaç: Coronaviruslar ve Rotaviruslar, Türkiye’de ve Dünya’da hayvan ve insan sağlığını tehdit eden önemli virolojik etkenlerdir. Sığırlarda görülen Bovine Rotavirus (BRV) ve Bovine Coronavirus (BCoV) önemli ekonomik kayıplara yol açmaktadır. Bu çalışmada sığırlar ile aynı çiftlikte bulunan Anadolu mandalarında BRV ve BCoV varlığının teşhis edilmesi amaçlanmıştır. Bu amaçla iki virusun varlığı Revers Transkriptaz Polimeraz Zincir Reaksiyonu (RT-PCR) ve BRV-BCoV hızlı testi ile tespit edildi. Bu iki testin sensitivite ve spesifite oranları karşılaştırıldı. Gereç ve Yöntem: Bu çalışmada Afyonkarahisar bölgesindeki sığır çiftliklerinde bulunan 230 adet Anadolu mandası klinik olarak değerlendirildi. Yirmi yedi adet klinik belirti (zayıflık, dehidrasyon, kusma, sulu kıvam ve sarı renkli dışkı) gösteren mandadan gaita numuneleri toplandı. Gaita numuneleri BRV ve BCoV yönünden Hızlı test ve RT-PCR ile değerlendirildi. Belirtilen analizler, kullanılan ticari kitlerin prosedürüne uygun olarak gerçekleştirildi. Bulgular: RT-PCR sonucunda %22.2 (27/6) BRV ve % 3.7 (27/1) oranında BCoV pozitiflik tespit edilirken, Rota-Corona Hızlı testle tüm numuneler negatif belirlendi. Her iki etken için rapid testin RT-PCR ile karşılaştırıldığında sensitivite ve spesifitesi sırayla %0 ve %100 olarak belirlendi. Ayrıca analiz edilen örneklerde istatistiki olarak BRV pozitiflik oranı, BCoV’a göre daha önemli bulundu (p<0,05). Öneri: Sonuç olarak rapid test sensitivitesinin düşük çıkması, enterik enfeksiyonların seyri boyunca saçılan virus miktarındaki değişikliğe bağlı olabilir.Öğe Expression of DNA methyltransferases (DNMTs) at mRNA level in ovine endometrium during estrus cycle and early pregnancy(Selçuk Üniversitesi, 2022) Hitit, Mustafa; Köse, MehmetAim: To elucidate mRNA expression of DNA methyltransferase enzyme genes (DNMT1, DNMT3A, and DNMT3B) in ovine endometrium whether they are modulated during the estrus cycle and early pregnancy. Materials and Methods: The endometrial samples including intercaruncular sites was obtained from a total of 24 ewes on days of 12 (P12, n = 4), 16 (P16, n = 4) and 22 (P22, n = 4) of pregnancy following mating and cyclic days of 12 (C12, n = 4), 16 (C16, n = 4) and 22 (C22, n = 4) of the estrous cycle. The relative mRNA level of DNMTs were evaluated through real-time quantitative RT-qPCR. Results: Abundances of DNMTs including DNMT1, DNMT3A, and DNMT3B were detected during the estrous cycle and early pregnancy in the ovine endometrium by this study. DNMT1 mRNA steady-state level was greater in P16 than in C16 whereas they did not change within the rest of the groups. The level of DNMT3A mRNA, during early pregnancy, had the highest expression levels on P12 compared to P22 and P16 (p<0.01). However, the level of DNMT3A mRNA was lower in P16 than in C16 (p<0.01) and had a similar decrease in P22 compared to C22 (p<0.01). DNMT3B mRNA level did not differ (p>0.01) during the estrus cycle and early pregnancy. Conclusion: DNA methylation related DNMTs may be required to control of gene expression in the ovine endometrium during the estrus cycle and early pregnancy.Öğe The expression of steroidogenic genes in ovine corpus luteum during early pregnancy(Selçuk Üniversitesi, 2021) Koçak, Nadir; Hitit, Mustafa; Köse, Mehmet; Bülbül, Bülent; Atlı, Mehmet OsmanAim: The goal of this study was to investigate the expression of steroidogenic genes in ovine corpus luteum during early pregnancy Materials and Methods: The animal model was designed as pregnancy; ewes were divided into three sub-groups, pregnancy 12th day: P12, pregnancy 16th day: P16, pregnancy 22th day: P22, and cyclic day 16 (C16). The expression of steroidogenic genes (steroidogenic acute regulatory protein; StAR, cytochrome P450 side-chain cleavage; P450sscc, and 3b-hydroxysteroid dehydrogenase/delta5 delta4-isomerase; 3?HSD) was evaluated using qPCR, and mRNA localization of StAR was detected on P16 against C16 through in-situ hybridization. Results: The expression of StAR mRNA was higher on day P22 and P16 compared to P12 (p<0.05), while it was at a steady-state level on day P22 vs P16 (p>0,05). mRNA expression of P450scc was greater on day P22 and P16 than day P12 (p<0.05), however on day P22 vs P16, it was at a steady-state level (p>0,05). Also, mRNA expression of 3?HSD had a similar trend; it was higher on day P22 and P16 compared to P12 (p<0.05), while it was at a steady-state level on day P22 vs P16 (p>0,05). In in-situ hybridization, we did not detect StAR mRNA on cyclic day 16 (C16) while abundantly expressed in luteal cells in P16. Conclusion: The mRNA expression of steroidogenic genes may appear to play a critical role during early pregnancy in ewes. Accordingly, it can be suggested that the steroidogenic pathway in the corpus luteum of ewe may transcriptionally regulate progesterone synthesis required for maintenance of early pregnancy.Öğe The expression of steroidogenic genes in ovine corpus luteum during early pregnancy(Selçuk Üniversitesi Veterinerlik Fakültesi, 2021) Hitit, Mustafa; Köse,Mehmet; Bülbül, Bülent; Atlı, Mehmet Osman; Koçak, NadirAmaç: Bu çalışmanın amacı erken gebelikte koyun korpus luteumunda steroidojenik genlerinin ekspresyonunu araştırmaktır. Gereç ve Yöntem: Çalışmanın hayvan modelinde gebe koyunlar üç alt grup olacak şekilde dizayn edilmiştir; gebelik 12: G12, gebelik 16: G16 ve gebelik 22: G22. gün ve siklik 16. gün (S16). Steroidojenik genlerin ekspresyon seviyesi (StAR, 3?HSD, P450scc) kantitatif polimeraz zincir reaksiyonu (PZR) ile değerlendirilmiştir ve StAR mRNA lokalizasyonu gebelik 16. günde siklik 16. güne karşı in situ hibridizasyon yöntemi ile gösterilmiştir. Bulgular: Gebelik günlerinde (G16 ve G22) StAR mRNA ekspresyonu G12 ile kıyaslandığında daha fazla olmasına (p<0.05) karşın, G16 ve G22 günleri arasında fark bulunmamaktadır (p>0.05). Benzer olarak, P450scc mRNA ekspresyon seviyesi gebelik günleri G22 ve G16’da G12’den fazla olarak bulunmuştur (p<0.05). Ancak, P450scc mRNA ekspresyonunda G16 ve G22 günleri arasında fark bulunmamaktadır (p>0.05). Benzer olarak, 3?HSD mRNA ekspresyon seviyesi gebelik günleri G22 ve G16’da G12’den daha fazladır (p<0.05). Ancak, 3?HSD mRNA ekspresyonunda G16 ve G22 günleri arasında değişiklik tespit edilememiştir (p>0.05). Bu çalışmadan, in situ hibridizasyonda, S16 günde StAR mRNA’sı luteal hücrelerde bulunamadı. Buna rağmen, luteal hücrelerde G16 günde StAR mRNA tespit edildi. Öneri: Steroidojenik yolakta bulunan genlerin ekspresyon seviyesinin koyunlarda erken gebelikte kritik bir rol oynadığı görülmüştür. Sonuç olarak, koyunların korpus luteumundaki steroidojenik yolağın, erken gebeliğin devamlılığı için gerekli progesteron sentezini transkripsiyonel olarak düzenleyebileceği önerilebilir.Öğe Investigation of interferon-tau stimulated genes (ISGs) simultaneously in the endometrium, corpus luteum (CL) and peripheral blood leukocytes (PBLs) in the preluteolytic stage of early pregnancy in ewes(ELSEVIER SCIENCE BV, 2016) Kiyma, Zekeriya; Kose, Mehmet; Atli, Mehmet Osman; Ozel, Caglayan; Hitit, Mustafa; Sen, Gonca; Kaya, MuhammetThe aim of the present study was to investigate the expression profiles of Interferon-tau (IFN-T) stimulated genes (ISGs) at the mid-luteal stage of the cycle (on day 13) in the uterine endometrium and extra-uterine tissues, such as the corpus luteum (CL), and in the peripheral blood leukocytes (PBLs) of pregnant and non -pregnant ewes, The objective was to evaluate the possibility of using the regulation of ISG expression in PBLs as a possible early pregnancy indicator in ruminants. For this purpose, multiparous ewes were synchronized and either allowed to mate (pregnancy group) or detected in estrus (cyclic group; day 0). The ewes were slaughtered on day 13, and the PBLs, endometrium and luteal tissues were collected. Total RNA was isolated from eight cyclic and eight pregnant ewes, and qPCR was employed to detect the steady state levels of Interferon -stimulated gene 15 (ISG15), Myxovirus (influenza virus) resistance 1 (Mxl) and Receptor transporter protein 4 (RTP4) mRNAs. The expressions of ISG15, Mxl and RTP4 were detected in the endometrium, CL and PBLs on day 13 of the estrous cycle and pregnancy. The expressions of these ISGs were upregulated only in the endometrium of pregnant ewes compared to non-pregnant ewes, but this stimulation was not observed in the CL and PBLs. The results suggest that the embryo stimulates ISGs only in the endometrium, and the effects are not evident in the extra-uterine tissues on day 13 of pregnancy. This study suggests that the measurement of the ISG expression in the PBLs is not a reliable detection method of early pregnancy in ewes, which are in the preluteolytic stage of early pregnancy. (C) 2016 Elsevier B.V. All rights reserved.Öğe Luteal regresyonda liver X reseptör yolağının araştırılması(Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü, 2017) Hitit, Mustafa; Koçak, NadirKolesterol steroid hormonları, safra asidi ve vitamin D'nin öncüsü olan nötral bir lipittir. Liver X reseptörleri kolesterolün reverz taşımında (efluks) önemli rol üstlenir ve makrofaj gibi periferal hücrelerden kolesterolün dışarı atılmasını sağlar. LXR'nin karaciğer ve diğer organlardaki rolü açıklanmış olsada, LXR' nin üreme organlarındaki rolü moleküler düzeyde tam olarak bilinmemektedir. Korpus luteumda (KL) gebelik gerçekleşmemişse progesteron üretimi durur, hücre bütünlüğü kaybolur, doku kitlesi azalarak apoptozis ve otofaji yolakları vasıtasıyla regresyon başlar. Regresyon süresince meydana gelen progesteron azalışında kolesterolün luteal dokudan efluksunun rolü olup olmadığı netlik kazanmamıştır. Bu çalışmanın amacı, kolesterolün reverz olarak tranpsortunun luteal regresyona katkısının araştırılmasıdır. Bu amaç için iki araştırma yapıldı: 1) Birinci araştırmada, siklik (S12 ve S16-doğal luteolizis) ve gebelik (G12, G16 ve G22) gruplarından, 2) İkinci araştırmada siklusun 12. günde luteolizis PGF2α enjeksiyonu ile uyarılıp 1 saat (PG1, n=4), 4 saat (PG4, n=4), 16 saat (PG16, n=4) ve kontrol (n=4) olmak üzere KL doku örnekleri toplandı. LXR yolağı (LXRα, LXRβ, ABCA1, ABCG1, APOE ve APOA1), steroidogenik progesteron sentez yolağı (StAR, 3βHSD ve P450scc) ile kolesterol alım reseptörlerinin (SRBI ve LDLR) gen ifadeleri mRNA düzeyinde RT-qPZ ve LXR yolağının genlerinin protein seviyesi western blot ile araştırıldı. Ayrıca, in-situ hibridizasyon ile StAR (S16 vs G16), SR-BI (S12 vs PG1 vs PG4 vs PG16), LXRα, LXRβ, ABCA1 genlerinin mRNA lokalizasyonları saptandı. Doğal luteolizisde LXRα ve ABCA1 mRNA ekspresyonu değişmezken (P>0,05), LXRβ, ABCG1, APOE ve ApoA-1 mRNA ekspresyon seviyesi S16'da S12'ye göre artmıştır (P<0,05). Ancak, StAR, 3βHSD, P450scc LDLR ve SR-BI mRNA ekspresyonları azalmıştır (P<0,05). PGF2α indüklenmiş luteolizis PG1'de S12'ye göre LXRα, ABCA1 ve 3βHSD mRNA ekspresyonu azalırken (P<0,05), LXRβ ekspresyonu artmıştır (P<0,05). Ancak, StAR, P450scc, LDRL ve SR-BI mRNA ekspresyonları değişmemiştir (P>0,05). PG4'de S12'ye göre LXRα, LXRβ, APOE, ApoA-I, LDLR, SR-BI, StAR, P450scc ve 3βHSD mRNA ekspresyonu azalırken (P<0,05), ABCA1 ve ABCG1 mRNA ekspresyonu değişmemiştir (P>0,05). PG16'da S12'ye göre StAR, P450scc, 3βHSD, LDLR, SR-BI ve ApoA-I azalmış (P<0,05), ABCG1 ekspresyonu artmıştır (P<0,05). Doğal luteolizis S16'da S12'ye göre LXRβ protein seviyesi artarken (P<0,05), diğer genlerin protein seviyesi doğal ve indüklenmiş luteolizis gruplarında değişmemiştir (P>0,05). In-situ hibridizasyonda StAR mRNA'sının ifadesi G16'da S16'ya göre daha fazladır (P<0,05). SR-BI mRNA'sı PG1'de S12'ye göre değişmezken, PG4 ve PG16'ı luteal doku lokalizasyonlarında azalmıştır (P<0,05). Sonuç olarak, bu öngörüler doğrultusunda, kolesterolün reverz taşınımından ziyade kolesterolün lipoprotein aracılı olarak alımının azalmasının luteal regresyonun sebebi olabileceği düşünülmektedir.Öğe MikroRNA biyogenezi(2015) Hitit, Mustafa; Kurar, Ercan; Güzeloğlu, AydınÖz: MikroRNAlar (miRNA) kodlama yapmayan 21-24 nükleotid uzunluğunda RNA molekülleridir. Genel olarak translasyonun baskılanmasına veya mRNAnın yıkımlanmasına neden olurlar. MikroRNA ilk keşfedildiğinde solucanlarda olağandışı spesifik gen ekspresyon mekanizması olarak düşünülmesine rağmen, artık günümüzde ökaryotlarda önemli gen ekspresyon düzenleyicisi olarak kabul edilmektedir. MikroRNA biyogenezi çekirdekte RNA polimeraz II aracılığında transkripsiyon ile başlar ve hairpin yapısında olgun miRNA dizisini içeren uzun miRNA (pri-miRNA)dan oluşur. Hairpin yapısı Drosha (RNAaz III enzimi) ve kofaktörü DiGeorge kritik sendrom bölgesi 8 (DGCR8)den oluşan mikroprosesör tarafından kesilir. Oluşan prekürsör miRNA (pre-miRNA) nükleustan Exportin-5 ile sitoplazmaya taşınır ve diğer RNAaz III enzimi olan Dicer tarafından 21-24 nükleotid uzunluğundaki dubleks miRNAya kesilir. Olgun diziye kesilecek olan iplik miRNA, RNA indüklenmiş susturma kompleksinde (RISC) Argonautea yüklenir. MikroRNAnın 2-8 nükleotidlik çekirdek dizisi hedef mRNA ile tam olarak eşlendiğinde mRNAnın destabilizasyonu sağlanır. Ancak tam olarak eşlenmediği zaman translasyonal baskılanmaya neden olur. MikroRNAların gelişim, farklılaşma ve diğer fizyolojik fonksiyonlarda önemli rol aldığı gösterilmesine rağmen, düzensiz ifadesi durumunda farklı patolojik olaylar ile ilişkilendirilmiştir. MikroRNA biyogenezinin farklı fizyolojik süreçlerde ve hastalıklarda epigenetik etkisinin moleküler düzeyde anlaşılmasının potansiyel önemi bulunmaktadır.Öğe Pentoxifylline May Restore Kanamycin-Induced Renal Damage in Rats(UNIV FED RIO GRANDE DO SUL, 2018) Corum, Orhan; Ozdemir, Ozgur; Hitit, Mustafa; Corum, Duygu Durna; Coskun, Devran; Er, AyseBackground: Kidney damage can be caused by many factors, such as using certain drugs in high doses or over the long term. The use of one such group of drugs, aminoglycosides, which act as Gram-negative antibacterial therapeutic agents, can lead to nephrotoxicity. It has been hypothesized that aminoglycoside-induced nephrotoxicity might be prevented by using pentoxifylline, which has antioxidant and anti-inflammatory effects and improves microcirculation. The objective of this present research was to determine the protective effects of pentoxifylline on kanamycin-induced kidney damage. Materials, Methods & Results: Thirty-two male Wistar rats were divided into four groups as follows: control, pentoxifylline, kanamycin, and kanamycin + pentoxifylline. The control group received intraperitoneal (IP) injections of 0.5 mL normal saline solution once a day (d) (SID) for 20 d; the pentoxifylline group received IP injections of 50 mg/kg pentoxifylline twice a day (BID) for 20 d, the kanamycin group received subcutaneous (SC) injections of 500 mg/kg kanamycin SID for 20 d, and the kanamycin + pentoxifylline group received both SC injections of 500 mg/kg kanamycin SID and IP injections of 50 mg/kg pentoxifylline BID for 20 d. At the end of 20 d, blood samples were taken from the heart by cardiac puncture under general anesthesia. After euthanizing the rats by cervical dislocation under anesthesia, the kidneys were immediately removed, relative kidney weights were calculated, and routine pathologic evaluations were conducted. Hemogram parameters were measured using a blood cell count apparatus and serum biochemical parameters were measured using an autoanalyzer. Kanamycin also caused (P < 0.05) tubular degeneration and tubular dilatation. Although pentoxifylline significantly reduced the level of kanamycin-induced tubular degeneration (P < 0.05), it did not significantly reduce tubular dilatation. Increases in relative kidney weights (P < 0.05) and in interstitial mononuclear cell (MNC) infiltrates were observed in the kanamycin and kanamycin + pentoxifylline groups compared to those in the control and pentoxifylline groups. Statistically significant changes were determined in the levels of some hemogram and biochemical parameters within reference ranges (P < 0.05). Discussion: In this study, both tubular degeneration and dilatation were observed in the kanamycin group. Pentoxifylline inhibited (P < 0.05) kanamycin-induced tubular degeneration and appeared to also reduce tubular dilatation, although this reduction was not significant. Tubular necrosis, epithelial edema of proximal tubules, tubular fibrosis, and perivascular inflammation might also be observed in aminoglycoside-induced nephrotoxicity. In current research, pentoxifylline prevented tubular damage induced by kanamycin, but did not inhibit infiltration by MNCs. Pentoxifylline also ameliorated amikacin-or gentamycin-induced histopathologic changes, especially those associated with tubular structures. The protective effects of pentoxifylline on kanamycin-induced tubular nephrotoxicity in this research might be a result of its stimulating the production of prostaglandin, a vasodilator, and of its improving microcirculation. Although the anti-inflammatory effects of pentoxifylline have been reported, these did not inhibit kanamycin-induced infiltration by interstitial MNCs in the present study. These results could indicate that the anti-inflammatory effects of pentoxifylline are not obvious and/or are dose dependent. Statistically significantly changes were determined in the levels of some hemogram and biochemical parameters in reference ranges. However, these changes were within the reference ranges for rats. These results suggested that kanamycin-induced tubular degeneration and dilatation might be prevented by administering pentoxifylline.Öğe Selection of reliable reference genes for qRT-PCR analysis on head and neck squamous cell carcinomas(Scientific Publishers of India, 2017) Yiğin, Aysel Kalaycı; Çora, Tülin; Acar, Hasan; Kurar, Ercan; Kayış, S. A.; Çolpan, Bahar; Öztürk, Kayhan; Hitit, MustafaThe choice of reliable reference genes as an internal control is inevitable to obtain accurate results. Here we present an assessment of 7 reference genes (18S rRNA, 28S rRNA, ACTB, GAPDH, TUBA1, YWHAZ, and SDHA) to normalize gene expression data in Head and Neck Squamous Cell Carcinomas (HNSCCs). We attempted to determine a reliable set of reference genes to use in the normalization of gene expression data in Head and Neck Squamous Cell Carcinomas (HNSCCs) and normal mucosal tissues. Malignant and non-malignant tissue samples were collected from 12 patients with primary untreated HNSCC. geNorm and NormFinder software packages were used for data evaluations. Results obtained by geNorm indicated that average expression stability values (M) of all candidates genes were smaller than 1.5 (accepted M value for geNorm), showing that all the evaluated genes can be employed as HKGs, although GAPDH and ACTB were reported to be the most stable. Similarly, NormFinder results were in agreement with geNorm’s results. GAPDH and ACTB are considered to be most suitable reference genes to evaluate novel gene expression in the tissues several of HNSCCs. © 2017, Scientific Publishers of India. All rights reserved.