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Item Isolation and characterization of a new type of aerobic, oxalic acid utilizing bacteria, and proposal of Oxalicibacterium flavum gen. nov., sp. nov.(Elsevier GmbH, 2002) Tamer A.Ü.; Aragno M.; Şahin N.A mesophilic, aerobic oxalic acid utilizing yellow-pigmented bacterium has been isolated from litter of oxalate producing plants in the region of Izmir (Turkey). It is motile by means of 1-3 polar flagella. Optimal growth occurred between 25-30 °C at pH 6,9. The G+C content of DNA is 62-64 mol % (Tm). Based on its morphological and biochemical features the organism belongs to the genus Pseudomonas, but differs from all the previously described species. The taxonomic relationships among strains described as or previously tentatively assigned to the genus Pseudomonas were investigated using numerical classification, DNA base composition and DNA-DNA hybridization. 16S rDNA sequences were determined for the strain TA17. On the basis of 16S rDNA sequence comparisons, physiological and biochemical characteristics, it is proposed to classify TA17T in a new genus and species for which the name Oxalicibacterium flavum gen. nov., sp. nov. is proposed. The type strain is TA17T (= NEU98T, = LMG 21571T).Item Immunolocalization of αV, α3 and β1 integrins in the human placenta with pre-eclampsia(Elsevier GmbH, 2003) Vatansever H.S.; Inan V.S.; Lacin S.; Koyuncu F.Signs of pre-eclampsia are considered to be caused by maternal endothelial dysfunction due to circulating factors of placental origin. Integrins are a large family of cell surface proteins that serve as receptors involved in cell-cell and cell-matrix interactions during placentation. Therefore, low expression of integrins or the lack of it may be encountered during pre-eclampsia. In the present study, we investigated the immunolocalisation of integrins αV, α3 and β1 in placentas of normal and preeclamptic women. Thirty-two placentas from pre-eclamptic (n = 14) and normotensive (n = 18) women were used. Immunohistochemical staining was performed on formalin-fixed, paraffin-embedded tissue specimens, using anti-αV, anti-α3 and anti-β1 antibodies and the indirect immunoperoxidase technique. A semi-quantitative grading system (HSCORE) was used to compare immunohistochemical staining intensities. Distribution patterns of αV, α3 and β1 integrins were detected in cytotrophoblasts and Hofbauer cells in normal and pre-eclamptic placentas. Immunostaining of αV and β1 integrins was slightly decreased in pre-eclamptic samples but α3 integrin immunostaining was similar in pre-eclamptic and normal placentas. Decreased immunostaining of integrins in the cytotrophoblasts may considered to be a structural basis for decreased placental perfusion in pre-eclampsia.Item Immunolocalization of integrins and fibronectin in tubal pregnancy(Elsevier GmbH, 2004) Inan S.; Giray G.; Vatansever H.S.; Ozbilgin K.; Kuscu N.K.; Sayhan S.Integrins are a large family of cell adhesion molecules that serve as receptors involved in cell-to-cell and cell-to-matrix interactions during implantation. We studied immunohistochemical staining of integrins (α3, αV, β1, and α2β1) and fibronectin in ectopic tubal pregnancy. Thirty fallopian tube samples with ectopic pregnancies and five normal tubal segments were obtained during ligation operations; the latter specimens served as controls in the study. Formalin-fixed paraffin-embedded tissue sections were stained with hematoxylin-eosin or primary antibodies against α3, β1, αV, and α2β1 integrins and fibronectin, using the avidin-biotin-peroxidase method. A semi-quantitative grading system was used to compare staining intensities. In the control samples, immunostaining of all integrins was found in a single layer of tall columnar epithelial cells, the lamina propria (Lp) and the muscular layer. Fibronectin staining was detected in the Lp and the muscular layer. Staining intensities of α3 and β1 integrins and fibronectin were increased in the normal part of fallopian tubes with ectopic pregnancies. Staining of β1 integrin was more intense than staining of α3 and fibronectin, whereas there was no difference in αV and α2β1 integrin expression between normal tubal tissue in the ectopic pregnancy group and control tubal tissue. In the tubal pregnancy group at the site of implantation, staining intensity of α3 and β1 integrins and fibronectin was strong in decidual cells, supporting tissue and placental villi, whereas αV and α2β1 staining was mild. We concluded that integrins, especially β1 and α3, and fibronectin may play a role in progression of tubal implantation. Although the role of integrins has not yet been clearly defined, these molecules may function as markers of normal and abnormal states of receptivity. We like to suggest that integrins and fibronectin, which are needed in utero implantation, are expressed in tubal tissues during ectopic pregnancy and are involved in ectopic implantation. © 2004 Elsevier GmbH. All rights reserved.Item Histopathological effects of sildenafil citrate on rat corpus cavernosum(Elsevier GmbH, 2004) Gümüs B.; Vatansever H.S.; Müezzinoǧlu T.; Müftüoǧluc S.; Kaymaz F.; Büyüksu C.Sildenafil citrate (Viagra) is widely used for the treatment of erectile dysfunction with various etiologies. The aim of the present study was the investigation of histopathological effects of sildenafil citrate on rat corpus cavernosum using light and electron microscopical techniques. Twenty male rats were divided into two groups. The first group (n = 10) was used as a control and the second group (n = 10) was treated with sildenafil citrate. Penile tissue was collected, fixed with formalin and embedded in paraffin for light microscopy, or fixed with gluteraldehyde and osmium tetroxide and embedded in Epon for electron microscopy. Light microscopical analysis showed that the corpus cavernosum was elongated and the number of blood vessels was increased. The amount of connective tissue in the penis was increased and dense collagen and smooth muscle fibers were observed in treated rats. Electron microscopical analysis showed that stromal structures of the corpus cavernosum (collagen fibers and number of cellular elements) were increased in treated rats. Fibroblasts showed signs of activation and the number of other stromaL cells was increased. Immature newly synthesized collagen fibers were observed and penetrated endothelial basement membranes. In addition, endothelial cells also showed signs of activation such as cytoplasmic granules in treated rats, whereas the surface area of blood vessels was increased and basement membranes were thickened. These histopathological changes due to treatment with sildenafil citrate indicate that prolonged use of sildenafil citrate may increase the risk of fibrosis in the penis. © 2003 Elsevier GmbH. All rights reserved.Item The effect of spermatic vessel ligation on testicular nitric oxide levels and germ cell-specific apoptosis in rat testis(Elsevier GmbH, 2005) Taneli F.; Vatansever S.; Ulman C.; Yilmaz O.; Giray G.; Genç A.; Taneli C.Management of high testis may vary but the most popular method in surgical treatment is the Fowler-Stephens maneuver. The aim of the present study was to investigate the effects of spermatic vessel ligation on testicular nitric oxide (NO) levels, expression of inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS) and germ cell-specific apoptosis in both ipsilateral and contralateral testes in rats. Twenty-eight animals were randomly allocated into four groups (n=7 each). The spermatic vessels were ligated as a simulation of the Fowler-Stephens maneuver. The groups of animals were sacrificed at 2 h (group 1), 4 h (group 2) and 24 h (group 3) after ligation, respectively. Sham-operated animals served as controls (group 4). Biochemical assessment of testicular NO levels was performed by the Griess method. iNOS and eNOS expression and apoptosis were studied in ipsilateral and contralateral testes. Testicular NO levels at 24 h after the simulated Fowler-Stephens maneuver were found to be significantly increased in both ipsilateral and contralateral testes when compared with the sham-operated group. eNOS expression was clearly increased in ipsilateral testes, whereas moderate expression was detected in the contralateral seminiferous tubules at 24 h after ligation. Mild focal iNOS immunostaining was also observed in seminiferous tubules of the ipsilateral testis at 24 h after the simulated Fowler-Stephens maneuver. Apoptosis was dramatically increased in ipsilateral testes; however, it was only detected in single cells in the contralateral side at 24 h after ligation. In conclusion, the simulated Fowler-Stephens maneuver induces testicular nitric oxide synthesis and germ cell-specific apoptosis in the ipsilateral testis. These results suggest that high levels of NO induce apoptosis and may impair spermatogenesis thus explaining the unsuccessful outcome of the Fowler-Stephens maneuver. © 2004 Elsevier GmbH. All rights reserved.Item Changes in distribution patterns of integrins in endometrium in copper T380 intrauterine device users(Elsevier GmbH, 2005) Oruç S.; Vatansever H.S.; Karaer Ö.; Eskicioǧlu F.; Narlikuyu B.Intrauterine contraception is the most cost-effective reversible method of contraception today, but its mechanism of action is not well understood. Our objective was to investigate immunohistochemical distribution patterns of αv, α3, β1 integrins in women using a copper T380 intrauterine device (IUD) for different periods of time to obtain insight into the role of integrins in intrauterine contraception. Endometrial biopsies were obtained from patients using T Cu380A IUD in follicular and luteal phases and in menopausal women grouped according to the period of time of IUD use (group 1: <3 year, and group 2: ≥3 years). Each group consisted of 10 patients, with a total number of 60 patients. Labelling intensity of all integrins, except for β1 which increased in the follicular phase, were decreased in women who used IUD for ≥3 years when compared with group 1 in the follicular and luteal phases and in the menopause. We conclude that long-term use of IUD affects integrin expression in endometrium not only in follicular and luteal phases of premenopausal women but also in postmenopausal women. Copper IUD can inhibit binding of integrins to the extracellular matrix and it may cause inhibition of the implantation stage, which is crucial for pregnancy. © 2005 Elsevier GmbH. All rights reserved.Item Changed Bcl:Bax ratio in endometrium of patients with unexplained infertility(Elsevier GmbH, 2005) Vatansever H.S.; Lacin S.; Ozbilgin M.K.Apoptosis has been shown to be an important regulator of endometrial function during the menstrual cycle and implantation. Recently, some possible implantation defects were identified in patients with unexplained infertility. In this study, we investigated the role of spontaneous apoptosis, which is regulated by death regulatory genes, such as Bcl-2, Bax, p53, and isoenzymes of nitric oxide synthases; eNOS and iNOS during the implantation window in women with unexplained infertility. Endometrial samples were evaluated from fertile (n=15) and unexplained-infertile women (n=15) during post-ovulatory 7th or 8th day of their menstrual cycles. Apoptotic cells were detected using the dUTP nick-end labelling assay and Bcl-2, Bax, p53, iNOS and eNOS were assessed immunohistochemically. Reduced apoptotic cells, weak immunoreactivity of p53 and strong immunoreactivity of Bcl-2 were observed in the unexplained-infertile group compared with the fertile group (p<0.001). Bax intensity was similar in both groups. While weak iNOS immunoreactivity was detected in both groups, moderately increased eNOS immunoreactivity was observed in infertile cases. Spontaneous apoptosis is reduced in the endometrium of unexplained-infertile women, and is associated with the changed Bcl-2:Bax ratio. This finding may be a contributing factor to defective implantation causing infertility in this group of patients. © 2005 Elsevier GmbH. All rights reserved.Item Histopathological and ultrastructural effects of Losartan on embryonic rat kidney(Elsevier GmbH, 2005) Akil I.; Inan S.; Gurcu B.; Nazikoglu A.; Ozbilgin K.; Muftuoglu S.The aim of our study was to investigate the histopathological, immunohistochemical and ultrastructural effects of Losartan (a selective angiotensin II type-1 receptor blocker) on renal development in rats. Twelve pregnant rats were divided into control and experimental groups. In the experimental group, Losartan (10 mg/kg/day) was given via nasogastric tube, between the sixth day of implantation and time of sacrifice on embryonic days 18 and 20. All formalin-fixed, paraffin wax-embedded renal tissue sections were stained with hematoxylin and eosin or labelled for binding of primary antibodies against transforming growth factor-β (TGF-β 1,-2,-3) using an avidin-biotin-peroxidase method. For electron microscopic examination, samples were fixed with glutaraldehyde and osmium tetroxide and embedded in araldite. Glomerular basement membrane (GBM) thickness was measured and compared using an unpaired t-test. Angiotensin II type-1 receptor antagonism by Losartan inhibited renal growth and delayed nephron maturation. Increased immunoreactivity of TGF-β's was observed in developing nephron precursors and interstitial cells in the experimental group. Electron microscopical examination showed that thickening of the GBM was normal in the control group but an irregular thickening was seen in the experimental group (p<0.001). It was also seen that epithelial cells of developing tubules underwent apoptosis in the experimental group. Thus, renal development in rats seems to depend on an intact renin-angiotensin system. © 2005 Elsevier GmbH. All rights reserved.Item The role of ATP sensitive K+ channels and of nitric oxide synthase on myocardial ischemia/reperfusion-induced apoptosis(Elsevier GmbH, 2006) Gok S.; Vatansever S.; Vural K.; Sekuri C.; Izanli A.; Tezcan A.; Cilaker S.During ischemia, ATP-sensitive K+ channels (KATP channels) open, and this triggers necrotic processes and apoptosis. In this study, we investigated whether selective sarcoplasmic and mitochondrial KATP channel blockers affected myocardial apoptosis and nitric oxide synthase (NOS) activity in a rat model of myocardial ischemia/reperfusion in vitro. Isolated rat hearts were subjected to 30 min of coronary artery occlusion followed by 30 min of reperfusion. A selective sarcKATP channel blocker, HMR1098 and a selective mitoKATP channel blocker, 5-hydroxydecanoate, were added to the perfusion fluid 10 min before occlusion. Myocardial apoptosis was detected immunohistochemically using the TUNEL method. Myocardial inducible NOS (iNOS) and endothelial NOS (eNOS) were determined immunohistochemically. In control hearts, apoptosis induction was associated with a greater immunoreactivity of iNOS than eNOS. Treatment with HMR1098, at a concentration of 3 μmol/l, significantly reduced the TUNEL-positive cardiomyocytes and this was associated with decreased iNOS and increased eNOS immunoreactivity. When this drug was administered at a higher concentration, at 30 μmol/l, a more marked reduction in apoptosis was observed but, in contrast to the effects observed at the lower drug concentration, eNOS immunoreactivity was almost completely abolished while iNOS was strong. Moreover, ischemia-induced cardiac dysfunction (e.g. contractile force and recovery of coronary flow) was increased by the higher concentration of HMR 1098. In hearts treated with 5-hydroxydecanoate, myocyte apoptosis was slightly reduced, and this was associated with an almost equal increase in both iNOS and eNOS immunoreactivity. These findings suggest that iNOS appears to be more important than eNOS in the reduction of apoptosis. However, the further inhibition of apoptosis by the higher concentration of HMR 1098 was associated with poorer cardiac function. © 2006 Elsevier GmbH. All rights reserved.Item Determination of the complex permittivity of materials with transmission/reflection measurements in rectangular waveguides(Elsevier GmbH, 2006) Şimşek S.; Işik C.; Topuz E.; Esen B.Determination of the complex permittivity of materials via transmission/reflection measurement in rectangular waveguides is formulated as an inverse scattering problem. The presented approach is directly applicable to measurements performed with various sample sizes and orientations. The predicted results are compared with measurements and good agreement is obtained. © 2006 Elsevier GmbH. All rights reserved.Item Characterization of osteoblasts derived from bone marrow stromal cells in a modified cell culture system(Elsevier GmbH, 2006) Deliloglu-Gurhan S.I.; Vatansever H.S.; Ozdal-Kurt F.; Tuglu I.Bone marrow is a complex tissue composed of hematopoietic and stromal stem cells with the potential to differentiate into adipogenic, fibroblastic, reticular, osteogenic and chondrogenic lineages. Identification of differentiation markers during transformation of stromal cells into osteoblasts in a time-dependent manner may be informative for cell-based tissue engineering. Therefore, we investigated the effects of osteogenic medium (OM) on the proliferation and differentiation of rat bone marrow stromal cells (BMSCs). BMSCs from adult male rat tibia and femur were collected and cultured in α-MEM medium with 10% fetal bovine serum, penicillin, streptomycin and gentamycin. After three days of culture, the medium covering the adherent cells in culture was changed to OM containing dexamethasone, Na-β-glycerophosphate and ascorbic acid. As a control, cell culture was also continued in the original medium for the same time period. Differentiated osteoblast cells were collected after 7, 10, 14, 21 and 30 days of culture, fixed with 4% paraformaldehyde and their immunolabelling for osteoblast markers osteonectin (ON) and osteocalcin (OC) was assessed using an indirect immunoperoxidase technique. Immunoabelling of ON and OC was detectable from day 10 of culture, began to increase on day 14, and increased steadily through to day 21. Labelling was highest on day 30 and was more intense in cells cultured with OM compared to the culture without OM. The control cells cultured in the absence of OM produced negligible levels of both markers. In conclusion, our culture system facilitated differentiation of BMSCs into osteoblasts featuring osteoblast markers, and these cells may be useful in autologous bone implant for the treatment of bone wound healing. © 2005 Elsevier GmbH. All rights reserved.Item Histophysiological effects of fluid resuscitation on heart, lung and brain tissues in rats with hypovolemia(Elsevier GmbH, 2006) Ekerbicer N.; Inan S.; Tarakci F.; Cilaker S.; Ozbek M.The efficacy of using colloids and crystalloids in the treatment of hypovolemia still remains controversial. An important aspect in treating hypovolemia is to re-establish normal tissue hemodynamics after fluid resuscitation. Production of nitric oxide (NO) or growth factors such as transforming growth factor beta (TGF-β) has been identified as a key mechanism in physiological and pathological processes in the different systems. This study was designed to investigate the histophysiological effects of resuscitation with different plasma substitutes on the heart, lung and brain tissues following acute blood loss in male Sprague-Dawley rats weighing 250-280 g (n=30). After anesthesia with sodium pentobarbital, the left femoral vein and artery were cannulated for the administration of volume expanders and for direct measurement of arterial pressure and heart rate. Twenty rats were bled (5 ml/10 min) and infused (5 ml/10 min) with one of four randomly selected solutions, (a) human albumin, (b) gelatin (Gelofusine), (c) dextran-70 (Macrodex); or (d) physiological saline (0.9% isotonic saline). Five control rats were bled without infusion. Tissue samples were taken and fixed in 10% formalin solution, then processed for embedding in paraffin wax. Sections were cut and stained with hematoxylin and eosin. Indirect immunohistochemical labelling was performed to reveal binding of primary antibodies against endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS) and TGF-β. Mild immunoreactivity of eNOS was observed in endothelial cells of vessels in brain, heart and lung tissues. Increased immunoreactivities of eNOS, iNOS and TGF-β were observed in the non-fluid resuscitated group in these organs; mild, moderate, moderate and strong immunoreactivities were seen in the albumin, gelatin, physiological saline and dextran-70 treated groups, respectively. Immunoreactivities of iNOS and TGF-β in the non-fluid resuscitated group were increased significantly, in comparison to the other groups, apart from the dextran-70 treated group. The results of this study show that gelatin solution and physiological saline may be of use after acute blood loss, and dextran-70 is not the preferred resuscitation fluid in the early stages of acute blood loss. It was concluded that albumin solution is the preferred fluid for resuscitation. © 2006 Elsevier GmbH. All rights reserved.Item Histopathological and ultrastructural effects of glycolic acid on rat skin(Elsevier GmbH, 2006) Inan S.; Oztukcan S.; Vatansever S.; Turel Ermertcan A.; Zeybek D.; Oksal A.; Giray G.; Muftuoglu S.Use of alpha hydroxy acids (AHA) to ameliorate specific dermatological problems with keratinization has become fairly widespread. The aim of this study was to evaluate the effects of the AHA derivative of glycolic acid, applied in different dosages, on rat skin using light and electron microscopy. Skin biopsies were taken from the dorsal side of rats ( n = 16) and at the end of each week after applying solutions containing AHA: week 1, 8% ( n = 5); week 2, 50% ( n = 5); week 3, 70% ( n = 6). The skin samples were fixed in 10% formalin for histology and 2.5% glutaraldehyde solution for electron microscopy and processed using routine protocols. Histological sections were stained with hematoxylin and eosin (H&E), Masson's trichrome and were also labelled for binding of a primary antibody against collagen I using the avidin-biotin-peroxidase method. The epidermal thicknesses were measured and the fibroblast count of the dermis was taken and the results compared using the statistical ANOVA test. Semi-thin sections were stained with toluidine blue-azure II solution and ultrathin sections were contrasted with uranyl acetate and lead citrate. Histochemical and immunohistochemical observations demonstrated that AHA treatment resulted in statistically significant increased thickness of the epidermis and an increase in numbers of active fibroblasts and in the amount of dense collagen, especially at higher dosages of AHA. Ultrastructural examination of rat skin from AHA-treated groups showed cytoplasmic vacuolization in epidermal keratinocytes, intercellular dysjunctions, and increased quantities of organized bundles of collagen fibers in the dermis. The use of AHA in appropriate dosages has been found to play an important role in the treatment of specific skin disorders, however, the harmful effects of use of AHAs at higher concentrations should not be ignored. We conclude that alpha hydroxyl acids have a wide spectrum of use in the field of dermatology but, due to side-effects, their use, dosage, and time frame should be restricted to the advice of dermatologists. © 2006 Elsevier GmbH. All rights reserved.Item The effect of Misoprostol, a prostaglandin E1 analog, on apoptosis in ischemia-reperfusion-induced intestinal injury(Elsevier GmbH, 2007) Topcu I.; Vatansever S.; Var A.; Cavus Z.; Cilaker S.; Sakarya M.The aim of this study was to investigate whether Misoprostol, a synthetic prostaglandin (PG) E1 analog, has any effect on the prevention of apoptosis in ischemia-reperfusion (I/R)-induced intestinal injury. Thirty adult male Wistar albino rats were divided into three groups: group I=sham operated+saline; group II=I/R+saline; and group III=I/R+Misoprostol. Misoprostol (50 μg/kg/d) was administered as an intragastric meal for 3 days. The terminal ileum was collected for histological and biochemical investigations. Apoptotic cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelled (TUNEL) reaction. Immunohistochemical analysis was performed to determine the distribution of inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS). Samples were also analyzed for malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). The number of TUNEL-positive cells was higher in group II when compared to the other two groups (p<0.05). In group III this value was higher when compared to group I, but lower than group II (p<0.05). iNOS immunoreactivity was not detected in ileum sections of group I animals, but moderate immunoreactivity was seen in group II and mild immunoreactivity in group III. The immunoreactivity of eNOS was moderate in ileum sections of all three groups. In ileum tissue, MDA was found to be higher in group II compared to group I (p<0.05), but there was no difference in group III. SOD was not different between groups I and III, but was significantly higher in group II (p<0.05). In our experimental model of I/R-induced intestinal injury, apoptosis is induced in enterocytes, whereas Misoprostol decreases enterocyte apoptosis in this experimental model. Our results indicate that Misoprostol may play a key role in the pathophysiologic events leading to failure of the intrinsic gut barrier defense mechanisms of intestinal epithelium. © 2007 Elsevier GmbH. All rights reserved.Item Significance of tyrosine kinase activity on malign transformation of ovarian tumors: A comparison between EGF-R and TGF-α(Elsevier GmbH, 2008) Zeren T.; Inan S.; Seda Vatansever H.; Ekerbicer N.; Sayhan S.Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-α) are members of the polypeptide growth factor family. The epidermal growth factor-receptor (EGF-R) is a receptor tyrosine kinase of the ErbB family. Many types of cancer, including ovarian cancer, display enhanced EGF-R immunoreactivity on their cell surface membranes. Also, an increase in TGF-α synthesis and secretion usually occurs in human carcinoma cell lines. In this study, we compared the immunoreactivities of TGF-α and EGF-R in ovarian tumors and related immunohistochemical findings to the histological type of the tumors. Formalin-fixed, paraffin wax-embedded tissue sections from 40 patients who had serous-mucinous borderline tumor and serous-mucinous adenocarcinoma of the ovary (n=10 each) were stained with hematoxylin-eosin and labeled for binding of primary antibodies against TGF-α and EGF-R using an avidin-biotin-peroxidase method. A semi-quantitative grading system was used to compare immunohistochemical labeling intensities. Increased immunoreactivity of EGF-R and moderate immunoreactivity of TGF-α was detected in adenocarcinomas. There was no significant difference in the immunoreactivity of TGF-α among the histologic types of ovarian tumors. The results of this study support the hypothesis that EGF-R may be a more useful marker than TGF-α in epithelial ovarian tumors. © 2007 Elsevier GmbH. All rights reserved.Item Cell surface glycosylation diversity of embryonic thymic tissues(Elsevier GmbH, 2008) Balcan E.; Tuǧlu I.; Şahin M.; Toparlak P.In the thymus, glycosylation status of many cell surface molecules changes during the thymocyte maturation and selection processes. In this study, we evaluated the glycosylation changes and possible relationships with programmed cell death in the thymic tissues from mouse embryos at the days 14 (E14), 15 (E15), 16 (E16), 17 (E17) and 18 (E18) of embryonic development. In order to determine glycosylation changes we used three different plant lectins: peanut agglutinin (PNA), Maackia amurensis leucoagglutinin (MAL or MAAI) and Sambucus nigra agglutinin (SNA), which recognize core disaccharide galactose (1-3) N-acetylgalactosamine [Galβ(1→3)GalNAc], sialic acid linked (2→3) to galactose [SAα(2→3)Gal] and sialic acid linked to galactose [SAα(2→6)Gal] structures, respectively. Our lectin histochemistry and lectin blotting studies indicated that glycosylation pattern was modified in thymocytes at the embryonic developmental stages analyzed. The immature cortical thymocytes were labeled by PNA, whereas medullary thymocytes were positive for MAL and SNA binding. Many medullary thymocytes exhibited α(2→6)-linked sialic acid on their surface and this increased throughout the gestational stages. In the lectin blotting studies, different protein bands of various molecular weights were identified in thymocytes. Two of them were putatively identified as CD43 and CD45 glycoproteins. In addition, TUNEL (deoxynucleotdyltransferase-mediated dUDP nick end labeling) indicated that only PNA-positive cortical thymocytes were deleted in all embryonic stages. These results indicate that the glycosylation pattern was modified in thymocytes at all embryonic developmental stages, and these modifications can affect the T cell deletion, probably via the galectin-1 molecule in the embryonic thymus. © 2007 Elsevier GmbH. All rights reserved.Item The effects of Gemcitabine and Vinorelbine on inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) distribution of MCF-7 breast cancer cells(Elsevier GmbH, 2011) Zeybek N.D.; Inan S.; Ekerbicer N.; Vatansever H.S.; Karakaya J.; Muftuoglu S.F.Gemcitabine, which induces S-phase arrest, and Vinorelbine, which arrests microtubule organization, are two agents that have demonstrate preferred anti-tumor activity. Nitric oxide acts in diverse functions including anti-tumor and anti-pathogenic activities. In this study, we aimed to examine the distribution of immunoreactivities of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in cells of the MCF-7 breast cancer cell line in response to treatment with Gemcitabine (G), Vinorelbine (V) and combination of Gemcitabine and Vinorelbine (G+V). The distributions of iNOS and eNOS were determined by using indirect immunoperoxidase or immunofluorescence methods and ELISA. Cells incubated with G, V and G+V for 24, 48 and 72. h were immunolabelled with anti-eNOS and anti-iNOS primary antibodies. Apoptosis was determined by TUNEL assay. A significant increase of eNOS immunolabelling on MCF-7 cells treated with G and G+V was observed. Apoptotic cells were also detected in G, V and G+V treated MCF-7 cells. The immunolabelling of iNOS was detected in all groups but this immunoreactivity was not different among the groups. In conclusion, while G treatment, induced S-phase arrest, triggered the NOS pathway after treatment of MCF-7 cells, V treatment, arrested microtubule organization and did not change the NOS pathway. Detection of increased eNOS immunolabelling and apoptosis after G treatment of MCF-7 cells could be important to the treatment of human breast cancer. © 2009 Elsevier GmbH.Item Significance of apoptosis related proteins on malignant transformation of ovarian tumors: A comparison between Bcl-2/Bax ratio and p53 immunoreactivity(Elsevier GmbH, 2014) Zeren T.; Inan S.; Vatansever H.S.; Sayhan S.In this study, we compared the immunoreactivities of Bcl-2, Bax and p53 proteins in ovarian tumors and related the immunohistochemical findings to the histological type of the tumors. Formalin-fixed, paraffin wax-embedded tissue sections from 40 patients who had serous-mucinous borderline tumors and serous-mucinous adenocarcinoma of the ovary (. n=. 10 each) were stained with hematoxylin-eosin (H&E). After histopathological examination, serial sections were stained immunohistochemically with primary antibodies to Bcl-2, Bax and p53 using an avidin-biotin-peroxidase method. A semi-quantitative grading system was used to compare the immunohistochemical staining intensities. The nuclear DNA fragmentation of apoptosis was determined using TUNEL method. As a result of immunohistochemical staining, increased immunoreactivity of Bcl-2 was observed in adenocarcinomas when compared to borderline tumors (. P<. 0.001). Strong immunoreactivity of Bcl-2 and mild immunoreactivities of Bax and p53 were detected in ovarian adenocarcinomas. There were no significant statistical differences in the immunoreactivity of Bax among the histological type of ovarian tumors. Whereas a balance was observed between the immunoreactivities of Bcl-2 and Bax in the borderline cases, and this balance was strongly changed toward the anti-apoptotic Bcl-2 protein in patients with adenocarcinoma. TUNEL staining of sections indicated apoptotic cells in the serous borderline tumors were about 8-fold higher than in the serous adenocarcinoma. The results of this study on apoptosis-related factors might help to develop novel protective and therapeutic approaches, such as isoflavonoids and isothiocyanates, which were associated with decreased Bcl-2/Bax ratio, against the malignant epithelial ovarian tumors. © 2014 Elsevier GmbH.Item A digital measurement system based on laser displacement sensor for piezoelectric ceramic discs vibration characterization(Elsevier GmbH, 2016) Parali L.; Pechousek J.; Şabikoʇlu I.; Novak P.; Navarik J.; Vujtek M.This study describes the innovative design of a digital measurement system based on a laser displacement sensor (LDS) as a vibrometer which is capable to measure a dynamic displacement response dependence on a stimulated vibration. The frequency response of a piezoelectric ceramic disc is obtained by processing the input/output signals obtained from the function generator and digital oscilloscope (digitizer) cards driven by a personal computer. Resonant frequencies of vibration are achieved utilizing the swept-sine signal excitation following the peak values in the signal response measured by LDS. The analogue signal from LDS controller represents directly a mechanical vibration of a piezoceramic disc. The test measurement results indicate that the system can distinguish resonance frequencies of piezoelectric ceramic discs up to 40 kHz with the resolution 1 Hz. Piezoelectric coefficient d33 and its linearity along the excited voltage amplitudes have been calculated by the applied methods as a demonstration of a successful system concept. The results achieved are in compliance with the reference value declared by the manufacturer of the piezoceramic disc. © 2015 Elsevier GmbH. All rights reserved.Item Effects of chronic amiodarone treatment on rat testis(Elsevier GmbH, 2016) Özkaya A.K.; Dilber E.; Gürgen S.G.; Kutlu Ö.; Cansu A.; Gedik Y.Amiodarone is a potent agent used to treat tachyarrhythmias, which are especially refractory to other medications, in both adults and children. Although widely used as an antiarrhythmic drug, amiodarone causes many serious adverse effects that limit its use. This study investigated the possible morphological and apoptotic effects of amiodarone on rat testes. Amiodarone was administered to male Sprague-Dawley rats at doses of 20 or 200 mg/kg/day for 14 days. A histopathological examination of testicular tissue revealed the presence of inflammatory cells in the seminiferous tubule lumen together with swelling and vacuolization in the cytoplasm of some spermatogonia; these effects occured in a dose-dependent manner. Immunohistochemical staining showed evidence of apoptosis, including caspase-3, caspase-9, Bax and increased DNA fragmentation was detected via a terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. In conclusion, the results show that chronic amiodarone treatment causes dose-dependent degenerative and apoptotic effects on rat testes. © 2016 Elsevier GmbH.