Browsing by Author "Erdemli, Esra"

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    Both Granulocytic and Non-Granulocytic Blood Cells Are Affected in Patients with Severe Congenital Neutropenia and Their Non-Neutropenic Family Members: An Evaluation of Morphology, Function, and Cell Death
    (2018) Erdemli, Esra; Tıp Fakültesi
    Objective: To examine granulocytic and non-granulocytic cells in children with severe congenital neutropenia (SCN) and their non-neutropenic parents. Materials and methods: Fifteen patients with SCN and 21 non-neutropenic parents were evaluated for a) CD95, CD95 ligand, annexin V, propidium iodide, cell cycle, and lymphocyte subsets by flow cytometry; b) rapid cell senescence (of leukocytes) by senescence-associated β-galactosidase stain; c) aggregation tests by aggregometer; d) in vitro bleeding time by PFA-100 instrument; e) mepacrine-labeled dense granule number of thrombocytes by fluorescence microscope; and f) hematomorphology by light and electron microscope. HAX1, ELANE, G6PC3, CSF3R, and JAGN1 mutations associated with SCN were studied in patients and several parents. Results: Significant increase in apoptosis and secondary necrosis in monocytes, lymphocytes, and granulocytes of the patients and parents was detected, irrespective of the mutation type. CD95 and CD95 ligand results implied that apoptosis was non-CD95-mediated. Leukocytes of 25%, 12.5%, and 0% of patients, parents, and controls showed rapid cell senescence. The cell cycle analysis testable in four cases showed G1 arrest and apoptosis in lymphocytes of three. The patients had HAX1 (n=6), ELANE (n=2), G6PC3 (n=2), and unidentified (n=5) mutations. The CD3, CD4, and NK lymphocytes were below normal levels in 16.6%, 8.3%, and 36.4% of the patients and in 0%, 0%, and 15.4% of the parents (controls: 0%, 0%, 5.6%). The thrombocytes aggregated at low rates, dense granule number/thrombocyte ratio was low, and in vitro bleeding time was prolonged in 37.5%-66.6% of patients and 33.3%-63.2% of parents (vs. 0% in controls). Under electron and/or light microscope, the neutrophils, monocytes, lymphocytes, and thrombocytes in the peripheral blood of both patients and parents were dysplastic and the bone marrow of patients revealed increased phagocytic activity, dysmegakaryopoiesis, and necrotic and apoptotic cells. Ultrastructurally, thrombocyte adhesion, aggregation, and release were inadequate. Conclusion: In cases of SCN, patients’ pluripotent hematopoietic stem cells and their non-neutropenic parents are both affected irrespective of the genetic defect.
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    Erişkin fare ovaryumunda olası germline ve pluripotent kök hücrelerin araştırılması
    (Biyoteknoloji Enstitüsü, 2015) Esmaeılıan, Yashar; Erdemli, Esra; Biyoteknoloji
    According to the classical knowledge of biology, in the ovary of female mammals there are a limited number of oocytes and there is no possibility of renewal if the oocytes are lost due to disease or injury. However, in the last 10 years, the results of studies about renewal and formation of oocytes and follicles in the adult mammalian ovary are lead to changing of this view completely. The aim of this thesis is demonstration of presence of putative germline and pluripotent stem cells and investigates the possibility of neo-oogenesis in the adult mouse (8 week-old BALB/c) ovary. Immunoflorescence staining of ovarian tissue sections revealed the presence of pluripotent stem cell (Oct-4, Nanog and Sox2) and germline stem cell (DAZL and DDX4/VASA) markers in the cytoplasm and nucleus of oocytes in different follicular stages. Presence of these proteins was also observed in the nucleus and cytoplasm of the cells that harvested from ovarian cell culture by immunoflorescence staining. In the ovary cell culture, oocyte differentiation process led to formation of morphologically oocyte-like cells and in some of these cells parthenogenesis was occurred. As a result of differentiation process formation of follicle-like structures was also observed. In the ovarian tissue and non-differentiated cells that harvested from cell culture, expression of the Oct-4, Sox2, Nanog, DDX4 and DAZL genes was demonstrated by qRT-PCR method. Nevertheless, in the harvested cells comparing ovarian tissue, despite the increased expression of Nanog, expression of Oct-4, Sox2, DAZL and DDX4 was decreased significantly (p<0.05). In the cells that differentiated into oocyte, expression of oocyte specific genes and in the parthenogenesis occurred cells, expression of pluripotent specific genes was demonstrated by RT-PCR method. As a result of western blot analysis, expression of Oct-4, Sox2, Nanog, DDX4 and DAZL proteins was observed in the ovarian tissue and non-differentiated cells that harvested from cell culture. Based on our study outputs, presence of pluripotent and germline stem cells in the adult mouse ovary was demonstrated. Under in vitro circumstances, differentiation potential of these cells into oocyte-like cells, follicle-like structures and in some cases parthenogenesis-like phenomena was observed.
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    İnsan semen kriyoprezervasyonunda koenzim Q10 ve kurkuminin spermatozoon morfolojisine ve fonksiyonuna etkileri
    (Tıp Fakültesi, 2019) Taş, Derya Özdemir; Erdemli, Esra; Tıp
    Spermatozoonların sıvı azot buharındaki kriyoprezervasyonu diğer vitrifikasyon yöntemlerinden daha iyi sonuçlar vermesine rağmen hücrelerin hareketlilik, canlılık, akrozom bütünlüğü, DNA ve kromatin yapısında hasar oluşturmaktadır. Oluşan hasar, çiftlerde düşük fertilite oranlarına neden olmaktadır. Kriyoprezervasyondaki bu problemler araştırmacıları farklı sulandırıcı formulasyonların geliştirilmesine örneğin sulandırıcıya çeşitli hormon, vitamin, şeker, antioksidan gibi maddelerin ilave edilmesine yöneltmiştir. Buradan yola çıkarak tezimizde, insan semen örneklerine rutinde kullanılan dondurma medyumuna koenzim Q10 ve kurkumin antioksidanlarını eklemeyi, dondurma/çözme işlemi sonrası medyumların spermatozoonları nasıl etkilediğini değerlendirmeyi, böylece antioksidan moleküllerin insan spermatozoon kriyoprezervasyonundaki hasar oranlarındaki etkisini göstermeyi amaçladık. Ön deneylerle spermatozoonların kriyoprezervasyon sonrası canlılık ve hareketliliğinin kurkumin 25 mM ve koenzim Q10 25 µM konsantrasyonlarında daha iyi sonuçlar vermesi nedeniyle çalışmaya bu dozlarda devam edilmiştir. Maddeler kullanım kolaylığı açısından ve glisrolde eriyebilirilikleri nedeniyle kriyoprotektan içerisine katılmıştır. Spermatozoon kriyoprezervasyonunda kurkuminin, hücrelerin hareketlilik ve canlılık parametrelerini kontrol grubuyla benzer şekilde etkilediği tespit edilmiştir. İmmunfloresan ve akım sitometrisi sonucu kurkuminin ve K+CoQ10 gruplarının hücrelerin akrozom bütünlüğüne kontrole göre benzer korunma sağlarken, DNA fragmantasyonunda ciddi artış oluşturdukları gözlemlenmiştir. Koenzim Q10, spermatozoon canlılığını kontrol grubuna benzer şekilde etkilerken özellikle CASA (Computer-assisted sperm analysis) sonucu ileri hareketli spermatozoon yüzdesini anlamlı şekilde arttırmıştır. Akrozom bütünlüğü ve DNA fragmantasyonuna belirgin bir etkisi tespit edilmemiştir. Sonuç olarak bu çalışmada, insan semen kriyoprezervasyonunda kurkuminin kullanılması hücrelerde DNA hasarına yol açmaktayken koenzim Q10 spermatozoon ileri hareketliliğine olumlu yönde etki etmiştir. Antioksidanların birlikte kullanılması dondurma çözme işleminde artı bir koruyucu etki göstermemiştir. Aksine DNA hasarını arttırıştır. Literatürde kriyoprezervasyonda olumlu etkilerinin olduğu kurkuminin daha ileri araştırmalara ihtiyacı olduğu görülmektedir. Koenzim Q10'un ise hareketliliği arttırmasının nedeninin açıklanması açısından organel düzeyinde etkisinin araştırılmaya ihtiyacı vardır. Anahtar Sözcükler: İnsan spermatozoon hücresi, kriyoprezervasyon, antioksidan, kurkumin, koenzim Q10
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    Osteoporoza Yeni Çözüm
    Erdemli, Esra