Browsing by Author "Özbilgin, K"
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Item The role of angiogenic factors in first trimester pregnancy lossesEskicioglu, F; Özbilgin, K; Taskend, S; Önal, T; Gökmen, TObjective: A good blood supply towards the peri-implantation endometrium is an essential requirement for pregnancy. Intermedin (IMD), vascular endothelial growth factor (VEGF), and endothelial nitric oxide synthase (eNOS), are angiogenic and vasoactive agents that play contributory roles in endometrial vascularity. The goal of this study was to immunohistochemically investigate the roles of various vasoactive factors in first trimester pregnancy losses. Materials and Methods: This was a prospective case-controlled study carried out on decidual and placental tissue samples obtained from women with unwanted pregnancies who served as the control group (n=10), and those with missed abortions who were the missed abortion group(n = 10). Immunohistochemistry techniques were used to compare IMD, receptor activity modifying protein (RAMP)1, RAMP2, RAMP3, VEGF, and eNOS expression of decidual and placental cells. Immunostaining for these factors was evaluated semiquantitatively by H-score analysis. Results: IMD and RAMPs in decidual cells exhibited higher expression in the control group. However, IMD and RAM-22 had a stronger expression in placental cells in the missed abortion group. In the control group, VEGF and eNOS had a higher expression in decidual cells and on the placental side, especially in syncytiotrophoblasts and cytotrophoblasts. Conclusion: Expressions of vasoactive agents, such as IMD, VEGF, and eNOS, decrease in first trimester pregnancy losses. Additionally, a compensatory mechanism against decreased endometrial and subendometrial vascularity results in the death of the embryo/fetus enhances in missed abortion cases. This mechanism characterized by increased expressions of IMD and RAMP2 initially begins in the syncytiotrophoblasts and cytotrophoblasts.Item Effects of α-lactalbumin and sulindac on primary and metastatic human colon cancer cell linesAydemir, I; Vatansever, S; Özbilgin, KBackground: HAMLET (human alpha-lactalbumin made lethal to tumor cells) and sulindac are active ingredients which are used in the treatment of cancers. Under both in vivo and in vitro conditions, HAMLET binds to the surface of tumor cells and enters the cells. Sulindac is one of nonsteroidal anti-inflammatory drugs. It inhibits the growth of tumor cells by inducing the apoptosis. In our study, we aimed to investigate effects of alpha-lactalbumin and sulindac on COLO-320 primary and COLO-741 metastatic human colon carcinoma cell lines via mitochondrial apoptotic pathway by indirect immunocytochemistry and TUNEL assays. Materials and methods: The effects of alpha-lactalbumin and sulindac were assessed by using MTT assay at 24, 48 and 72 h and apoptosis markers caspase-3, caspase-9 and cytochrome-c were detected using immunocytochemistry and TUNEL methods. Results: It was appeared that alpha-lactalbumin and sulindac may triggger mechanisms of apoptosis in both primary and metastatic colon carcinoma cell lines and the primary colon carcinoma cell line was affected more than the metastatic cells. Conclusion: It is proposed that alpha-lactalbumin and sulindac can be used in cancer treatments and future in vivo experiments.Item Effects of alcohol during secondary neurulation in chick embryosMete, M; Aydemir, I; Ünsal, ÜÜ; Özbilgin, K; Tuglu, MI; Gürcü, BObjective: Alcohol continues to be consumed even though its harmful effecs are well established. One of the most common damage of alcohol consumption is fetal alcohol syndrome, characterized by craniofacial anomalies, cardiac anomalies and neural tube defects. Therefore, understanding the molecular mechanisms underlying the alcohol-induced toxicity that occur with time and dose dependent manner is very important., Most of the studies in order to understand the effects of alcohol have been carried out on early neurulation, however its effects on late neurulation are still unknown. Therefore in this study, effects of alcohol on secondary neurulation were investigated in chick embryos. Methods: Leghorn breed of embryonic chicken eggs were used. At 50 h of incubation, 100 mu L 50% ethanol solution was injected. Depending on the period of exposure to alcohol, varying degrees of pathological disorders were detected in E3, E7 and E10 days. Results: Developmental delay, structural abnormalities, morphological abnormalities in the heart and face and especially presence of two spinal cord cavities were found. In addition, we also detected delays in the closure of the neural tube, cellular deformities and the structural abnormalities in notochord. While eNOS, iNOS, and TUNEL levels increased, while laminin levels decreased. Conclusion: In this study during late development, significant alcohol-induced morphological and histopathological changes were observed. We also determined Increased level of oxidative stress caused by alcohol was accompanied with the changes in matrix composition. Better understanding of these mechanisms which affect the cell behavior is important and will allow learning of harmful effects of alcohol.Item The role of selectins in the first trimester pregnancy lossEskicioglu, F; Laçin, S; Özbilgin, K; Köse, CObjective: There are no well-defined findings about reasons for first trimester abortion in some pregnancy cases. Selectins are cell adhesion proteins which are important for blastocyst implantation in the decidua. The goal of the study was to investigate the role of selectins in first trimester pregnancy loss by immunohistochemistry Study design: Decidual and placental tissue samples have been obtained from the women with unwanted pregnancy as the control group (n=40) and missed abortion (n=40) as the study group. Immunohistochemistry technique has been used to compare P, L and E-selectin expression of the fibroblast and the decidual cells in uterine decidual stroma; and fibroblasts and mesenchymal cells in placental villous stroma. Immunostaining for P,L,E-Selectin has been evaluated semiquantitatively by HSCORE analysis. Results: Decidual cells, for E and L-selectin showed stronger staining in the study group than controls, and the difference was statistically significant (p = 0.001, p = 0.001). P-selectin showed stronger staining in the control group, but the difference was not as significant as the E and L-selectins (p=0.04). In the placenta, cytotrophoblasts and syncytiotrophoblasts showed stronger staining for P,E,L-selectins for the control group (p<0.001, p=0.001 and p<0.001, respectively). Conclusion: Strong expression of each of the three investigated selectins in healthy pregnancy villi shows their contribution to implantation and strong placentation. There is a need for better understanding of the functions of adhesive molecules in these events to reveal unknown causes for pregnancy loss.Item Effects of propranolol and paclitaxel on angiogenesis in breast cancer cell linesSimsek, F; Inan, S; Müftüoglu, S; Özbilgin, K; Vatansever, S; Tuglu, IPurpose: The aim of this study was to investigate the effect of propranolol (PR), which is used in infantile hemangiomas, and paclitaxel (PX), which is widely used as an chemotherapeutic agent, on cancer cells. Materials and Methods: That the cells counted with trypan blue the doubling time were determined. Also with MTT assay were analyzed the cytotoxic effect and IC50 value of drugs. In the breast cancer cell lines which are differents with regard to invasion (MDA-MB-231 and MCF-7) anti-VEGF, anti-eNOS, anti-iNOS and anti-ERK1/2 primer antibodies investigated by using immunohistochemical methods. To evaluation of immunoreactivity was used the H-scoring system. Results: With MTT test, IC50 values are applied to the cells dosage for MDA-MB-231 cells; PX: 5 nmol, PR: 50 mu m, and for MCF-7 cells PX: 3,7 nmol, PR: 50 mu m, were established. In immunohistochemical application, immunoreactivity of control group was increased with strong and/or stronger in the cancer cells, while those of in PX, PR and combine treatment was decreased either significant or very significant. Conclusion: With this study, application of anti chemotherapeutic therapy which is paclitaxel, in additon with anti angiogenic therapy in the treatment of breast cancer, vascular vasodilation, cell proliferation, migration, survival ultimately thought to be important in the prevention or reduce of angiogenesis.Item The maturity of intestinal neomucosaGünsar, C; Vatansever, HS; Arslan, OA; Sencan, A; Müftüoglu, S; Özbilgin, K; Kaymaz, F; Mir, EBackground/Purpose: The maturity of neomucosa growing on a serosal surface for the treatment of short bowel syndrome still is questionable. The aim of this study was to evaluate the intestinal neomucosa to assess its histologic maturity. Methods: A 6-cm-long isolated ileal segment (IS) was prepared in 8 Wistar albino-type rats. The IS was divided from the antimesenteric side, and 2 intestinal tubes were established, which shared a common wall and a common pedicle. After ileal biopsy sampling for the control group (CG), the IS was fashioned into a mucous fistula. Eight weeks later, all the rats were killed, and the ISs were investigated for neomucosal growth. Sections were prepared with periodic acid shift (PAS) and H & E staining for light microscopy. They also were evaluated by transmission electron microscopy. The microscopic morphology of the 2 groups was evaluated. Immunohistochemical staining was performed to show the expression of the tissue beta1, alpha3 and alpha2beta1 integrin subunits of both the neomucosa (NS) and control group (CG) segments. Results: Sections of the NS showed a well-arranged columnar epithelial cell layer with goblet cells that were generally located superficially and with a complete basement membrane. Under the electron microscope, the sections from the NS group showed an epithelial cell layer with proper microvilli of the same height, although they were shorter than those of the CG, and tight intercellular junctions between the epithelial cells. Significant differences between the NS and CG groups were found in the measurements of villus width at base, microvillus surface, and microvillus height. The lamina propria consisted of rich collagen fibers and active fibroblasts in the NS group. In the immunohistochemical staining, although beta1 integrine showed a dense distribution (+++) in the lamina propria, particularly localizing at the depth of the tunica mucosa layer, alpha3 integrin was observed to have a less dense immunoreactivity (++) in both groups. The expression of alpha2beta1 integrin showed slight and dispersed (+) staining. Conclusions: The NS showed histologic maturity and ultimate structural similarity with the native small bowel mucosa, which provides strong indirect evidence for the proper functioning of the neomucosa. (C) 2004 Elsevier Inc. All rights reserved.Item The aromatase inhibitor anastrozole is associated with favorable embryo development and implantation markers in mice ovarian stimulation cyclesKaraer, N; Vatansever, HS; Oruç, S; Özbilgin, K; Cilaker, S; Koyuncu, MFObjective: To investigate the embryonic and endometrial effects of anastrozole in preimplantation and implantation phases in FSH-induced cycles in mice. Design: Blind randomized study. Setting: University research laboratory. Animal(s): Twenty-seven mature female mice. Intervention(s): Single-dose anastrozole (25 mg/kg [0.75 mg]), recombinant FSH (5 IU/mL), and hCG (5 IU/mL) (n = 9); recombinant FSH (5 IU/mL) and hCG (5 IU/mL) (n = 9); or sterile saline (1 mL) (n = 9). The morning of finding the vaginal plug was designated as day 1 of embryonic development (E1). Three mice from each group were sacrificed on E1 and embryos aspirated from uterine tubes. The rest of the mice were sacrificed on E2.5-3 and uteruses removed. Main Outcome Measure(s): Embryo quality, endometrial histologic evaluation, and immunohistochemical analysis of tumor necrosis factor-alpha, leukemia inhibitory factor, laminin, and collagen IV staining. Result(s): Anastrozole use in FSH-induced cycles not only caused an increase in preimplantation receptivity and implantation but also supported release of implantation markers. The enhanced embryo development seen in this study would explain the higher implantation because embryo development is synchronized with endometrial development. Conclusion(s): In mice, the use of anastrozole in FSH-induced cycles has a positive effect on embryo quality and implantation. This effect might be species dependent, and human studies are needed. (c) 2005 by American Society for Reproductive Medicine.Item The effect of cardiopulmonary bypass on the expression of inducible nitric oxide synthase, endothelial nitric oxide synthase, and vascular endothelial growth factor in the internal mammary arteryToprak, V; Sirin, BH; Tok, D; Özbilgin, K; Saribülbül, OObjective: Endothelial function of a vessel may be impaired by local or systemic inflammation initiated by cardiopulmonary bypass (CPB) during coronary artery bypass graft (CABG) surgery. The present study was designed to investigate the early effects of CPB on nitric oxide production and vascular endothelial growth factor (VEGF) expression in internal mammary artery (IMA). Design: Prospective study. Setting: University hospital. Participants: Twenty patients who were scheduled for elective CABG with CPB. Interventions: IMA sections were studied immunohistochemically from these patients. The samples were taken from the distal end of the IMA before the institution of CPB and just before the construction of the IMA-left anterior descending artery anastomosis. Measurement and Main Results: After CPB, VEGF and endothelial nitric oxide synthase immunoreactivity increased significantly when compared with baseline values in the endothelium (p = 0.0156, p = 0.0313) and adventitia (p = 0.0313, p = 0.0001), respectively. No significant change was observed in inducible nitric oxide synthase immunoreactivity. Conclusions: The increase in eNOS expression may have been induced by the inflammation caused by CPB. (C) 2006 Elsevier Inc. All rights reserved.Item Analysis of the Effects of Inhibitor and Activator Systems (Smad's Proteins) of TGF-βs on Chick Neural Tube ClosureUmur, N; Vatansever, S; Umur, AS; Özbilgin, K; Selçuki, MThe families of TGF-beta s and Smads proteins that controls its intracellular signaling pathways are known to play a role in early neurulation. The aim of this study is to demonstrate distribution of TGF-beta s (1, 2, 3) and Smads (1/2/3, 6, 7) proteins as a system in different hours of neural tube development of chick embryos. The SPF eggs were incubated at 37.8 +/- 2 degrees C for 24(th), 30(th), 48(th), 72(nd) h. After that, embryos were examined using immunohistochemistry and western blotting techniques. To the results, TGF-beta s immunoreactivities (particularly TGF-beta 3) at the 24(th), 30(th) and 48(th) h of chick development (during neural tube closure) were determined and decreased at the 72nd h (after neural tube closure), but expressions of TGF-beta s were detected in all stage of embryos in western blotting. While Smad 1/2/3 immunoreactivitiy and expression was less than that of the Smad 6 and 7 at the 24(th), it was increased at the 30(th) h. Smads proteins immunoreactivities were decresead at the 72(nd) h. In conclusion, the members of TGF-beta s are play a role in chick neural tube closure, the secretions of TGF-beta s are controlled different Smad proteins. In addition, immunoblotting results showed that TGF-beta s and Smads proteins were effective in the development of all tissues and organs of the embryos.Item InsulinKuscu, NK; Koyuncu, F; Özbilgin, K; Inan, S; Tuglu, I; Karaer, ÖThe goal of this study was to investigate histological changes of the rat ovary treated with either insulin or insulin plus human chorionic gonadotropin (hCG). The study was conducted in Celal Bayar University, School of Medicine, Animal Research Laboratory. Eighteen adult female Wistar rats were divided into three groups to receive saline, or insulin, or insulin plus hCG for 4 weeks. At the end of treatment the rats were sacrificed and the ovaries were evaluated with hematoxylin and eosin. There was no abnormal change in. rats treated with saline. A thickened capsule, stromal hypertrophy and stromal cell hyperplasia, and no developing follicles, were observed in the insulin-only group. A thin capsule, developing follicles and corpora lutea, and normal theca cells and stroma were observed in the insulin-plus-hCG group. We conclude that insulin may lead to histological changes similar to stromal hyperthecosis and polycystic ovary syndrome, and may be one of the factors causing follicular arrest.Item Effects of verteporfin-mediated photodynamic therapy in breast cancer cellsSönmez, PK; Turhan, A; Öztatlici, M; Özbilgin, KPhotodynamic therapy works with a photosensitizer that is stimulated when exposed to a light source of a specific wavelength and produces a form of oxygen that can be used in cancer treatments. In this study, we investigated the effect of laser on apoptosis on breast cancer cell lines (MDA-MB-231) treated with verteporfin in cell culture media. Verteporfin added MDA-MB-231 cells were incubated without light for 24 hours after applying laser light at a wavelength of 695 nm at an intensity of 50 J/cm2 at various times. Anti-proliferative effects were evaluated by immunoreactivity of anti-Bcl-2 and anti-Bax antibodies by immunocytochemical staining. When anti-Bax/Anti-Bcl-2 ratio are compared, the ratio of 1.5 in the control group cells decreases in short-term laser applications, while it approaches normal values in the 7th min after longterm laser application and reaches a very high value in the 9th min. Therefore, our results suggest that verteporfin-mediated PDT may be a potential combined therapy strategy against breast carcinoma by increasing apoptosis.Item The Differentiation of Neuronal Cells from Mouse Embryonic Stem CellsUmur, N; Vatansever, HS; Umur, AS; Türköz, E; Özbilgin, KWith new technologies emerging today, the importance of stem cells in the cell therapy of nervous system diseases is supported by recent studies. Therefore, the development of neuronal cell differentiation protocols from stem cells is of great importance. In our study, the differentiation of neuronal and neuroglial cells from mouse embryonic stem (ES) cell line and their analysis with neuronal cell markers are aimed. Mouse ES cells were differentiated to neurogenic series cells by adding N2 and bFGF to the culture medium on coated Fibronectin dishes. For the identification of differentiated cells, they were evaluated by light microscopy using immunhistochemistry techniques and by electron microscopy. Indirect immunohistochemical staining method was performed with SSEA-1 (mouse embriyonic stem cells marker), Nestin (neural precursor cells marker), beta III-Tubulin (neuronal cells marker), MAP-2 (neuronal cells marker), GFAP (astrocyte marker), and O4 (oligodendrocyte marker). After 1 week of differentiation of cells, immunoreactivities of SSEA-1 and Nestin were detected to be negative and moderate, respectively. After 2 weeks culture time, the differentiation was still continuing and especially positive immunoreactivities of beta-III Tubulin and MAP-2 and weak immunoreactivities of O4 and GFAP were supported neuronal differentiation. In conclusion, our results suggest that neuronal cell derived from mouse ES cells were differentiated particularly to neuron using N2+bFGF+fibronectin culture condition. Therefore, these differentiated cells may be used as a treatment method in degenerative diseases of the nervous system.Item Intraoperative evaluation of testicular vascularization and perfusion in rat testicles with indocyanine green (ICG)/near-infrared (NIR) fluorescent imaging after torsion-detorsion and reperfusionSencan, A; Tanriverdi, HI; Simsek, FB; Usta, IB; Üçöz, M; Özbilgin, KIntroduction The aim of this study was to evaluate testicular perfusion and vascularization with intraoperative ICG/NIR imaging in a testicular ischemia-reperfusion model and to investigate the effects of ICG on testicular tissue. Materials and methods 24 male rats were divided into four groups. In the ICG group, only ICG was given and images of the testicles were recorded with NIR camera. In the torsion group, the testicles were left in torsion for 4 h. ICG/NIR images were obtained after torsion and detorsion. In the reperfusion group, ICG/NIR images of the testicles were obtained at the 4(th) hour of reperfusion. After the procedures, testicles were collected and evaluated with histological, immunohistochemical examination and qRT-PCR. Results There was no histologically negative effect of ICG on testicular tissue. There was no testicular perfusion in the torsion group, but perfusion started after detorsion. At the 4th hour of reperfusion, testicular perfusion continued. TNF-a, IL-6, MCP-1 and caspase-3 immunoreactivity were found to be at low levels in the control and ICG groups, while high in the torsion and reperfusion groups (p < 0.05). In qRT-PCR, TNF-a, IL-6, MCP-1 and caspase-3 expressions were lower in the control and ICG groups, but higher in the torsion and reperfusion groups. Conclusion There was no histologically negative effect of ICG on testicles. The ICG/NIR imaging technique seems to be a feasible method in testicular torsion and may contribute to the surgeon in the intraoperative management of testicular torsion. In testicles that started to be perfused after detorsion, perfusion still continued at the 4th hour of reperfusion. Our next goal is to test whether testicles showing ICG fluorescence in during reperfusion maintain their viability for long term.Item A comparison of the molecular distribution of proangiogenic factors in endometrium of missed abortions and of voluntary first trimester termination casesÖzçakir, T; Turan, MA; Simsek, F; Atay, C; Vatansevers, S; Özbilgin, KObjective: The authors aimed to evaluate the angiogenic changes that occur in the cases with missed abortions compared with the voluntary termination of pregnancy as control group, with this controlled clinical study. Materials and Methods: The study included fifteen healthy volunteer women with unwanted pregnancy less than 10th gestational week in an academic research environment. The patients were 19 women between 6th and 11th gestational weeks diagnosed with missed abortion as the patient group. Immunohistochemistry was utilized to examine temporal and spatial expression of vascular endothelial growth factor (VEGF) and their two receptors: VEGF-Rl (Flt-1) and VEGF-R2 (Flk-1/KDR), and Trombospondin-1, eNOS, iNOS, and HIF-1 alpha in the both deciduas and placenta of the both groups. Results: This study discovered the significant difference (p < 0.005) between the groups of controlled and missed abortion in the decidual and placental cell components, and has put forward that thrombospondin and iNOS have an impact on abortion through antiangiogenic effect in cases of missed abortions. Conclusions: The potential role of molecules affecting angiogenesis in the etiology of missed abortion has been evaluated and the authors aimed for this to be a guide for studies on further treatments and on the prevention of the development of missed abortions.Item Effect of raloxifene and atorvastatin in atherosclerotic process in ovariectomized ratsDemir, BÇ; Uyar, Y; Özbilgin, K; Köse, CAim: The goal of this study was to investigate the combined effects of raloxifene and atorvastatin in aged ovariectomized rats during endothelial dysfunction and atherosclerotic process. Material and Methods: This study was conducted on 28 Wistar albino female rats randomly divided into four groups. All groups were ovariectomized and one group was kept as the control group (OVX). For four weeks, the remaining three groups were treated with the statin atorvastatin (OVX+AV), the selective estrogen receptor modulator raloxifene (OVX+RL), and both atorvastatin and raloxifene (OVX+RL+AV), respectively. At the end of the treatment period, all rats were sacrificed and thoracic aortas excised, and endothelial cells were immunohistochemically stained for markers in the atherosclerotic process, such as inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS), endothelin-1 (ET-1), monocyte chemotactic protein-1 (MCP-1), and tumor necrosis factor alpha (TNF-a). Results: Compared to the ovariectomized group, the iNOS level was significantly increased in the OVX+RL group (P = 0.002), but contrarily decreased in the groups OVX+AV (P = 0.002) and OVX+RL+AV (P = 0.002). eNOS levels in the groups OVX+AV (P = 0.002) and OVX+RL+AV (P = 0.002) were significantly lower than that in the OVX group. When compared to the OVX group, significant reductions in ET-1 and TNF-a levels were found in all treatment groups. A significant decrement in MCP-1 level was found in the OVX+AV group (P = 0.002). Conclusion: In aged ovariectomized rats, the administration of both raloxifene and atorvastatin significantly decreased the levels of ET-1 and TNF-a on endothelial cells. Combined treatment with these drugs shortly after menopause might play a potential preventive role in the early stages of atherosclerosis development.