Browsing by Author "Vatansever S."
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Item Clomiphene citrate does not affect the secretion of α3, αv and β1 integrin molecules during the implantation window in patients with unexplained infertility(Oxford University Press, 2001) Lacin S.; Vatansever S.; Kuscu N.K.; Koyuncu F.; Ozbilgin K.; Ceylan E.Background: The expression of integrin molecules on the endometrium suggest that certain integrins may participate in the cascade of molecular events leading to successful implantation. A prospective, controlled study was carried out to investigate the effect of clomiphene citrate (CC) on secretions of β1, α3 and αv integrin molecules in the endometrium of patients with unexplained infertility during the implantation window. Methods: A total of 40 endometrial samples was evaluated in both spontaneous (n = 13) and ensuing clomiphene-treated cycles (100 mg on days 5-9) and also from fertile women serving as controls (n = 14) during postovulatory 7th or 8th day of menstrual cycle. A semiquantitative grading system (H-score) was used to compare the immunohistochemical staining intensities. Endometrial thickness and serum oestradiol and progesterone concentrations were also measured on the day of sampling. Results: Staining of αv but not β1 and α3 integrins was significantly less intense in infertile cases than fertile control cases (1.42 ± 0.12 versus 2.21 ± 0.13 respectively, P = 0.012) and this was not restored to normal concentrations with treatment. Conclusions: Our study indicated that cc treatment significantly decreased the endometrial thickness and increased oestradiol and progesterone concentrations. However, secretion of αv, β1 and α3 integrin molecules, which might play a role in implantation, was not affected.Item Immunolocalization of transforming growth factor-β3 in pregnant human myometrium: Possible associations with labor(2001) Kuşcu N.K.; Laçin S.; Vatansever S.; Yildirim Y.; Var A.; Uyanik B.S.; Koyuncu F.Background. Transforming growth factor-β3 is a cytokine which is involved in cell growth regulation and differentiation, stimulation of extracellular matrix and modulation of immune responses. The goal of this study was to detect the presence of this cytokine in the myometrium of preterm and term, nonlaboring and laboring patients, and to measure serum levels of interleukin-1β (IL-1β), IL-6 and IL-8 before cesarean section. Methods. In this prospective study, we obtained samples of myometrium from the lower uterine segment during elective and emergency cesarean sections (term non-laboring, n=8; term laboring, n=7; preterm non-laboring, n=3; and preterm laboring, n=19) and stained for transforming growth factor-β3. Blood was also sampled from the same patients to determine IL-1β, IL-6 and IL-8 levels. Results. Different intensities of staining were detected in preterm laboring, term nonlaboring and term laboring groups, but there was no staining in preterm nonlaboring group. We also found a statistically significant difference in IL-6 levels between laboring and nonlaboring groups (p=0.028). Conclusion. Different intensities of TGF-β3 which appeared in different stages of myometrium made us consider that TGF-β3 might prepare myometrium to labor, and IL-6 was more important than the other interleukins in initiation of labor.Item Comparative morphological differences between umbilical cords from chronic hypertensive and preeclamptic pregnancies(2002) Inan S.; Sanci M.; Can D.; Vatansever S.; Oztekin O.; Tinar S.To compare morphological changes in the umbilical cords from chronic hypertensive and preeclamptic patients having normal or pathological umbilical artery Doppler ultrasonographic results. Umbilical cords from 34 normotensive, 31 chronic hypertensive and 70 preeclamptic women with normal and abnormal Doppler flow velocity waveforms (FVW) at 35 40 gestational weeks were studied. Morphological changes in the umbilical cords were examined on formalin-fixed, paraffin-embedded sections. The total umbilical cord area, total vessel area, and wall thickness of umbilical vessels were measured in systematic random samples using unbiased stereology methods. An ANOVA test was used for statistical analysis. In the chronic hypertensive and preeclamptic groups with normal Doppler FVW, the thickness of the umbilical cord vessels remained nearly constant, whereas both the total area and the lumen area were reduced. These changes correlate with the histopathological findings, suggesting a mainly vasoconstrictive effect. By contrast, analysis of the preeclamptic group with pathologic Doppler FVW showed a comparable reduction of all parameters of the umbilical cord. Histopathological findings were related to smaller, contracted smooth muscle cells of the vessel wall, which is suggestive of a predominant hypoplastic mechanism. As a result of reduced uteroplacental perfusion, fetal hypoxia and intrauterine growth retardation become un-avoidable in preeclampsia. The histopathological changes in the umbilical cord between the chronic hypertensive and preeclamptic patients depend on the Doppler results. In conclusion, the umbilical artery Doppler FVW indices provide good values for predicting intrauterine growth retardation in preeclamptic patients.Item Immunohistochemical staining of IGF-I, IGF-binding proteins-1 and -3, and transforming growth factor beta-3 in the umbilical cords of preeclamptic patients(2002) Inan S.; Vatansever S.; Kuscu N.K.; Laçin S.; Ozbilgin K.; Koyuncu F.Background. To detect the immunoreactivity of insulin-like growth factor-I, insulin-like growth factor-binding proteins-1 and -3 and transforming growth factor beta-3 in the umbilical cords of normal and preeclamptic patients. Methods. Umbilical cords were obtained from 15 normal and 15 preeclamptic patients. Immunoreactivities were determined using either indirect immunofluorescence or immunoperoxidase techniques on formalin-fixed, paraffin-embedded sections. Staining intensity was graded by a semiquantitative scoring method. The results were compared by Mann-Whitney U-test. Results. The umbilical cords were thinner and the vessels were hypoplastic in the preeclamptic group. Moderate staining intensity for insulin-like growth factor-I, insulin-like growth factor binding protein-1 and -3 and transforming growth factor-beta 3 was observed in normal patients. The preeclamptic group had mild and strong intensities for insulin-like growth factor-I and insulin-like growth factor binding protein-1, respectively, and intensity for insulin-like growth factor binding protein-3 did not change, but diffuse and increased intensity was observed for transforming growth factor-beta 3. Conclusion. Changes in the intensity of insulin-like growth factor-I and its major binding protein and the transformation of growth factor-beta 3 may play a role in the pathogenesis of preeclampsia by altering the structure and responsiveness of the umbilical cord.Item Increased vascular surface density in ovarian endometriosis(Parthenon Publishing Group Ltd, 2003) Inan S.; Kuscu N.K.; Vatansever S.; Ozbilgin K.; Koyuncu F.; Sayhan S.Our goal in this study was to investigate the presence of angiogenesis-related factors in endometriomas by evaluating their vascular surface densities. Thirty ovarian samples were included in the study. Of these, ten were histologically confirmed endometriomas, ten were ovarian specimens in the follicular phase and ten were ovarian specimens in the luteal phase, serving as controls. Histological specimens were immunostained for von Willebrand factor (vWF: factor VIII-related antigen) and CD34. The area with the highest microvessel density in endometriosis and in the normal ovary was evaluated by using an intercept grid. All microvessels in a specific field (x 100 magnification) were counted and vascular surface density was measured, as 164.01 ± 21.26 vs. 125.15 ± 11.28 and 117.44 ± 9.27 by using vWF, and as 172.97 ± 25.64 vs. 138.65 ± 32.21 and 120.34 ± 18.40 by using CD34 in endometriotic, follicular and luteal ovarian samples, respectively (p < 0.001). The mean vascular surface density was significantly higher in endometriosis than in the ovarian samples of the follicular phase or the luteal phase. No significant difference was seen between normal ovarian samples. Endometriosis was associated with angiogenic properties. Having demonstrated elevated angiogenic factors in endometriotic samples, we concluded that activation of angiogenesis might be a key factor in the pathogenesis of endometriosis.Item Ultrastructural effect of sildenafil citrate on corpus cavernosum and other genital organs in female rats(2003) Kilinç K.; Gündüz M.I.; Gümüs B.H.; Vatansever S.; Kaymaz F.Aim: To determine the ultrastructural effects of sildenafil on the female genital organs. Methods: Twenty female cycling Wistar albino rats weighing 250 ± 20 g were randomly divided into two groups of 10 each. Rats of one group were gavaged with 0.5 mg·kg-1·d-1 of sildenafil 3 days in a week for 4 weeks and the other served as the controls. After cessation of treatment animals were sacrificed by cervical dislocation under methoxyflurane anaesthesia. The clitoris, vagina, uterus and bartholin glands were taken at the estrous and were fixed with 10 % formalin solution for light microscopy and 2.5 % glutaraldehyde and osmic acid for electron microscopy. Results: Under the light microscope, the fibrocollageous tissue was found increased, the capillaries enlarged and the connecting tissue elements increased in the corpus cavernosum in the treated group. On electron microscopy, increased connective tissue, fibroblasts with notched nucleus, shorten immature collagen fibers without striation were seen. Abundant foldings and penetration with collagen bundles were observed in the basal membrane. Large connection complexes, especially gap junctions among the wide capillary endothelial cells were observed. Conclusion: There are evident histological changes due to sildenafil citrate in female rat corpus cavernosum. The clitoris and bartholin glands were the most effected organs. While the histopathological changes of clitoral tissue could be expected, an increase in the mass of bartholin gland was surprised.Item Is a filum terminale with a normal appearance really normal?(Springer Verlag, 2003) Selçuki M.; Vatansever S.; Inan S.; Erdemli E.; Baǧdatoǧlu C.; Polat A.Introduction: Tethered spinal cord is defined as a condition in which the conus medullaris ends at a level below the L1-2 intervertebral space. The spinal cord is considered to be tethered when there is a thick filum terminale or low-lying conus medullaris. It has also been reported that a normal level of the conus medullaris and normal thickness of the filum terminale do not mean that there is no cord tethering. Materials and methods: In this investigation, we examined 21 fila terminalia; 5 of them were taken from cadavers, and these were used as a control group (group 1; n/n), 8 from patients with a normal appearance of the filum terminale but with clinical symptoms (incontinence) and pathologic results of a urodynamic study (group 2; n/ab), and 8 from patients with an abnormal appearance of the filum terminale and with clinical symptoms (group 3; ab/ab). Interestingly, we found that while fila terminalia in the control group were made up mainly of collagen fibers, more connective tissue with dense collagen fibers, some hyalinization and dilated capillaries were noticed in the fila from group 2. Results: Our results suggest that these histological features may reflect a decreased elasticity within the filum terminale, resulting in a tethering effect on the lower conus in otherwise normal physiological conditions. Conclusion: These findings lead us to reconsider sectioning of the filum terminale in incontinent patients with normal results in radiological studies, whose condition is called "nonneurogenic neurogenic bladder".Item Tear function changes of postmenopausal women in response to hormone replacement therapy(2003) Kuscu N.K.; Toprak A.B.; Vatansever S.; Koyuncu F.M.; Guler C.Objective: The goal of this study was to search the effect of hormone replacement therapy (HRT) on tear function changes in postmenopausal women. Methods: Following initial ophtalmic evaluation and tear sample collection, the subjects were given daily 0.625 mg conjugated estrogen, and either continuous combined or cyclic 5 mg medroxyprogesterone acetate was added. Eye examination included visual acuity, slit-lamp examination, fundus examination, tonometry, Schirmer's test, and break up time (BUT) evaluation. Six months later, control examination was done and repeat tear samples were obtained. Tear immune globulin A (IgA) and lysozyme levels were measured by gel electrophoresis. The pictures of the bands were evaluated by digital image analysis with Scion Image program. Results: Conjunctival vascular congestion, laxity and corneal desquamation did not change before and after HRT (P>0.05). A significant improvement was noted in meibomian gland inflammation (P=0.034). We have not observed any significant difference in burning, foreign body sensation, and tearing (P>0.05). In addition, no significant difference was noted in BUT (P=0.370) and Schirmer's test values (P=0.271). Though both lysozyme and IgA levels were elevated following the therapy, only IgA levels increased significantly (P=0.04). Conclusion: HRT decreased meibomian gland inflammation and increased tear lysozyme and IgA levels in postmenopausal women. © 2002 Elsevier Science Ireland Ltd. All rights reserved.Item Neural tissue continues its maturation at the site of neural tube closure defects: Implications for prenatal intervention in human samples(2004) Selçuki M.; Vatansever S.; Inan S.; Sanci M.; Sayhan S.; Baǧdatoǧlu C.Objective: Our objective was to investigate the relation between the embryological development and neural tissue maturation at the site where the neural plate failed to form a neural tube. Material and methods: Samples from 15 aborted human fetuses with neural tube defects (NTD). All of the fetuses were between 20 and 25 gestational weeks old. Indicators of neural tissue maturation, formation of basal lamina, expression of integrins and neuron specific class III beta tubulin (tuj1) were investigated. To detect the adverse effects of the environment, if any, p53 and bcl-2 activity at both sites of the open and closed neural plate were investigated as well. Results: No difference was found in the expression of maturation-related molecules at the site of the neural plate that remained open compared with the site where the neural tube is normally formed. While high p53 activity was noted in neural tissue at the site of the neural tube defect, no such activity was detected in the neural tissue where the neural tube is normally formed. Conclusion: Our results suggested that maturation and differentiation of neural tissue continued regardless of the failure of neural tube closure. Therefore, the neurological deficits that are encountered in NTD patients should be related to secondary damage such as amnion fluid toxicity, uterus contractions, labor, etc. It seems valuable to save the neural plate before the negative effects of the environment renders the neural tissue functionless. © Springer-Verlag 2004.Item Effects of rofecoxib, a selective cyclooxygenase-2 inhibitor, on endothelial dysfunction, lipid peroxidation, and hepatocyte morphology in rats with sepsis-induced liver damage(2004) Kara E.; Var A.; Vatansever S.; Cilaker S.; Kaya Y.; Coşkun T.Sepsis remains a difficult problem for clinicians, with its systemic effects and high morbidity and mortality rates. The roles of oxidative stress, endothelial dysfunction, and lipid peroxidation in sepsis-induced organ damage are being investigated. The aim of this study was to investigate the effects of selective cyclooxygenase (COX)-2 inhibition on tissue lipid peroxidation, endothelial dysfunction, and hepatic cell morphology in a rat model of sepsis. Thirty rats with sepsis induced by cecal ligation and puncture were divided equally into 3 groups: treatment group (rofecoxib 1 mg/kg PO), control group (saline 1 mL PO), and sham group (sham surgery only). All the rats were sacrificed 1 day after sepsis induction. The livers were removed using a median laparotomy for histopathologic and biochemical analysis. Histomorphologic hepatic damage and lipid peroxidation were significantly reduced in the rofecoxib treatment group compared with the control group (P < 0.05 and P = 0.001, respectively). Endothelial nitric oxide synthase and inducible nitric oxide synthase staining of liver samples was statistically significantly reduced in the treatment group compared with the control group (both, P < 0.001). The hepatic nitric oxide level and malonyldialdehyde activity decreased significantly (P < 0.001 and P = 0.001, respectively) in the rofecoxib group compared with the control group. Hepatic myeloperoxidase activity was similar between the treatment and control groups. Further investigation of selective COX-2 inhibition as an alternate therapeutic choice for sepsis-induced hepatic damage should be considered. Copyright © 2004 Excerpta Medica, Inc.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 The long-term effect of mesh bioprosthesis in inguinal hernia repair on testicular nitric oxide metabolism and apoptosis in rat testis(2005) Taneli F.; Aydede H.; Vatansever S.; Ulman C.; Ari Z.; Uyanik B.S.Polypropylene mesh is the most widely used material in inguinal hernia repair. Although polypropylene mesh is known as an inert material, it is experimentally proven that mesh generates a chronic inflammatory tissue reaction. The aim of the present study was to investigate the long-term effects of polypropylene mesh material used in inguinal hernia operations on testicular function, testicular nitric oxide (NO) metabolism and germ cell-specific apoptosis in rats. The study comprised 40 male rats that were randomly allocated into two groups. In group 1, the left spermatic cord was elevated and a 0.5 × 1 cm polypropylene mesh was placed behind the left inguinal spermatic cord and group 2 consisted of the sham-operated controls. Blood samples were taken at 6 months preoperatively and postoperatively after to assess luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels for hormonal evaluation. Testicular NO was evaluated by the Griess method, apoptosis by a TUNEL method and inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) expressions by immunohistochemical staining. Mild (+) eNOS expression was observed in all specimens. Mild (+) iNOS expression was only detected in ipsilateral testis of the mesh-implanted study group. Apoptotic cells were not detected in any samples. We are of the opinion that long-term polypropylene mesh implantation has no effect on testicular hormonal function and only a limited effect on nitric oxide levels and this effect is not sufficient to cause apoptosis in testis that could lead to infertility. It seems that mesh implantation is a reliable method in inguinal hernia repair; however, further work is required by more sensitive methods to fully elucidate the potential testicular damage. Copyright © 2004 John Wiley & Sons, Ltd.Item Pre-ischemic administration of nitric oxide synthase inhibitors reduced germ cell apoptosis after spermatic vessel ligation in the rat testis(2005) Taneli F.; Vatansever S.; Ulman C.; Giray G.; Genc A.; Taneli C.Introduction: The Fowler-Stephens maneuver, a mode of spermatic vessel ligation, is a method of choice in the management of high testes. The aim of the present study is to investigate the effects of pre-ischemic administration of NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide (NO) synthase (NOS) inhibitor, in preventing testicular ischemic damage and germ cell-specific apoptosis in rats. Materials and Methods: 30 min before ligation of the spermatic vessels, L-NAME was administered intraperitoneally to a group of rats and saline was given to another group of rats (controls). Biochemical assessments of NO and germ cell-specific apoptosis were performed. Results: Testicular NO levels in the L-NAME group showed significant decreases in the ipsilateral (p = 0.004) and contralateral (p = 0.015) testes in relation to those of the control group. The apoptotic indices were found in 2.3% of the L-NAME group and 3.1% of the control group. Conclusion: Pre-ischemic administrations of the NOS inhibitor, L-NAME, effectively decreased NO production and to some degree caused a reduction in germ cell apoptosis in the rat testes after spermatic vessel ligation. Further studies are mandatory to confirm our preliminary results and to address the potential introduction of NOS inhibitors into clinical practice. Copyright © 2005 S. Karger AG.Item Subcutaneous administration of live lactobacillus prevents sepsis-induced lung organ failure in rats(2005) Ilkgul O.; Aydede H.; Erhan Y.; Surucuoglu S.; Gazi H.; Vatansever S.; Taneli F.; Ulman C.; Kose C.; Bengmark S.The leading cause of death in intensive care units around the world is the syndrome of exaggerated and prolonged systemic inflammation response (SIRS), which if not controlled will lead to irreversible, often multiple, organ dysfunction and organ failure syndrome (MODS). Host phagocytic cells, predominantly macrophages and neutrophils, and their soluble products, play a central role. Accumulation of neutrophils is often observed, especially in the lungs, intestines, liver and kidneys. Neutrophilic infiltration of distant organs, and especially of the lungs, has been a characteristic finding of patients dying from sepsis. However, the mechanisms responsible for the exuberant inflammation and neutrophil infiltration are not yet fully understood. It has recently been demonstrated that this, at least in part, is due to an inhibition of neutrophil apoptosis, and there is a suggested association with activation of nuclear factor - κB (NF-κB), reduced activity of caspases-9 and -3 and maintenance of mitochondrial transmembrane potential.Item Computer assisted image analysis of peroxidase stained endometrial tissue(2006) Toprak A.B.; Vatansever S.; Özbilgin K.Aim: In immunohistochemistry, antibodies are used to detect and localize antigens in cellular compartments in sections taken from frozen or paraffin blocks. Currently, computerized image processing techniques are widely available to analyze immunohistochemically stained tissues; however, the image analysis programs are generally highly specific and expensive. We aimed to show the feasibility of computer assisted image analysis (CAIA) by using two computer programs currently in widespread use. Methods: Two available computer programs, Adobe Photoshop CS and ImageJ, were used to separate areas in the same tissue sections stained with NOS antibody and labeled with peroxidase in human endometrial tissue. CAIA was made on the relevant images using these two programs. Results: There was a significant difference among the groups according to staining intensity, in all three grades of staining (P < 0.001, P < 0.05, and P < 0.001, respectively). Subjective assessment of the images was in good agreement with objective CAIA. One (+) positive stained area was greater in group 1 and less in group 2 and group 3 (P < 0.001 and P < 0.001, respectively). Group 3, which was considered as the most intensely stained by the observer, had the greatest amount of ++ staining. Conclusion: CAIA is a safe and reliable method of analyzing tissue samples stained using immunohistochemical methods. © TÜBİTAK.Item Examination of bcl-2 and p53 expressions and apoptotic index by TUNEL method in psoriasis(2006) Gündüz K.; Demireli P.; Vatansever S.; Inanir I.Background: Psoriasis is characterised by hyperproliferation and by aberrant differentiation. Blockage of the normal apoptotic process is one of the factors implicated in the pathogenesis. Objective: To determine the apoptotic features by using TUNEL method and also bcl-2 and p53 expressions in psoriatic epidermis. Materials and methods: Biopsies of 35 patients with psoriasis vulgaris and 14 normal skin were evaluated. Apoptotic cells were detected using the dUTP nick-end labelling assay; bc1-2 and p53 expressions were assessed by using immunohistochemical techniques. A semi-quantitative grading system (HSCORE) was used for comparison. Results: Bcl-2 was strongly expressed in basal keratinocytes of normal skin, while no expression was observed in 13 (37.2%) of the psoriatic samples and it was weakly expressed in the spinous cell layer of 22 (62.8%) samples. Moderate p53 expression was observed in the psoriasis group, while it was weak in the control. The percentage of TUNEL positive cells were significantly increased in the psoriasis group (65±2.30) when compared with the control (32.84±7.16). Conclusion: Apoptotic index besides bc1-2 and p53 expressions in psoriasis differ from normal epidermis. Down-regulation of bc1-2 is consistent with the dynamics of psoriasis but increased TUNEL positive cells and p53 expression has not been fully elucidated yet. © Blackwell Munksgaard 2006.Item Immunolocalizations of VEGF, its receptors flt-1, KDR and TGF-β's in epithelial ovarian tumors(2006) Inan S.; Vatansever S.; Celik-Ozenci C.; Sanci M.; Dicle N.; Demir R.Objective: Angiogenesis is an essential factor for growth, differentiation, invasion and metastasis of tumors. In this study, we aimed to evaluate the immunolocalizations of vascular endothelial growth factor (VEGF), its receptors flt-1, KDR/flk-1, and transforming growth factor-beta's (TGF-β) in epithelial ovarian tumors, utilizing indirect immunohistochemistry to understand the role of the angiogenic events in ovarian neoplasia. Methods: Tissue blocks from 40 patients who had ovarian pathology (borderline serous-mucinous tumor and malignant serous-mucinous adenocarcinoma of the ovary) were included in this study. All formalin-fixed, paraffin-embedded tissue sections were stained with hematoxylin-eosin or primary antibodies against VEGF, flt-1, KDR/flk-1, TGF-β1, TGF-β2 and TGF-β3 using the avidin-biotin-peroxidase method. H-SCORE, a semi-quantitative grading system, was used to compare immunohistochemical staining intensities. Results: Positive VEGF immunoreactivity was concentrated in the epithelial and stromal parts of all the ovarian samples and the endothelial cells in the stroma were also stained. Increased immunoreactivity of VEGF was observed in malignant ovarian adenocarcinomas compared to the borderline tumors of the ovary. VEGF receptors, flt-1 and KDR/flk-1 immunoreactivities were detected not only in vascular endothelial cells, but also in tumor cells at malignant sites. Immunoreactivities of VEGF and its receptors were coexpressed in tumor cells of the ovarian carcinoma. While immunoreactivities of TGF-β1 and TGF-β2 were both overexpressed in malignant ovarian carcinomas, immunoreactivity of TGF-β3 was still mild. Conclusion: Our results suggest that overexpression of VEGF, its receptors flt-1, KDR/flk-1 and TGF-β interaction may play an important role in the ovarian cancer biology, with potential effects on tumor growth and angiogenesis. New therapeutic strategies using VEGF and TGF-β antagonists could obtain an additional approach to the treatment ovarian carcinoma by inhibiting angiogenesis.Item Chemotherapy influences inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) activity on 3D breast cancer cell line(Tech Science Press, 2006) Öktem G.; Bilir A.; Selvi N.; Yurtseven M.E.; Vatansever S.; Ates U.; Uysal A.; Omay S.B.Multicellular tumor spheroids (MTS) are three-dimensional structural forms of tumors grown in vitro in the laboratory. In this study, the aim was to determine the regulation of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) expressions on MTS in response to treatment with the commonly used anti-cancer drugs Doxorubicin and Docetaxel. The spheroids were generated using the "liquid overlay" technique. The distribution of both iNOS and eNOS was detected using indirect immunohistochemistry, while the expression of both iNOS and eNOS was measured using Western blots. Additionally, S-phase analysis using 5-bromo-2′- deoxyuridine (BrdU) was done on the MTS after treatment with doxorubicin, docetaxel, and a combination of the two. The Griess method was used to measure nitric oxide (NO) production in the cells. An increase in iNOS immunoreactivity and a decrease in eNOS immunoreactivity were observed after doxorubicin treatment, when compared with the other groups. Furthermore, upregulation of iNOS and downregulation of eNOS were detected in doxorubicin-treated cells using Western blotting. Insignificant iNOS expression was observed in all of the groups, and it was particularly low in the control and drug combination groups. NO production was also found to be significantly high after docetaxel treatment, and cell proliferation decreased after doxorubicin treatment. In conclusion, chemotherapy influences NOS activity differently with the presence of different drugs. The results with iNOS show that doxorubicin is a more effective drug than docetaxel, and a drug combination may play a helpful role in the suppression of tumorigenicity and cancer metastasis. Interestingly, eNOS expression increased after the addition of both docetaxel and the drug combination, and it was found to negatively correlate with the histological grade of the tumor. Therefore, analyzing the expression of both iNOS and eNOS might be very useful for targeting the treatment of breast carcinoma and obtaining better information on prognosis. Copyright © 2006 Cognizant Comm. Corp.Item Effect of apoptosis and response of extracellular matrix proteins after chemotherapy application on human breast cancer cell spheroids(Spandidos Publications, 2006) Oktem G.; Vatansever S.; Ayla S.; Uysal A.; Aktas S.; Karabulut B.; Bilir A.Multicellular Tumor Spheroid (MTS) represents a three-dimentional structural form of tumors in laboratory conditions, and it has the characteristics of avascular micrometastases or intervascular spaces of big tumors. Recent studies indicate that extracellular matrix (ECM) proteins play a critical role in tumor metastasis, therefore normal and cancer cells require an ECM for survival, proliferation and differentiation. Doxorubicin and Docetaxel are widely used in the therapy of breast cancer, as well as in in vivo and in vitro studies. In this study, we examined the effect of apoptosis and proliferation of cells on the human breast cancer cell line, MCF-7, by using p53, bcl-2 and Ki67 gene expression, and the tendency to metastasis with extracellular matrix proteins, laminin and type IV collagen after chemotherapy in the spheroid model. The apoptotic cell death in situ was detected by TUNEL method. TUNEL-positive cells and positive immunoreactivities of laminin, type IV collagen, p53 and, bcl-2 were detected in the control group. There was no laminin and type IV collagen immunoreactivities in spheroids of drug groups. While TUNEL-positive cells and p53 immunoreactivity were detected in Docetaxel, Doxorubicin and Docetaxel/Doxorubicin groups, p53 immunoreactivity was not observed in the Docetaxel group. There was no bcl-2 immunoreactivity in either drug group. In addition, we did not detect Ki67 immunoreactivity in both control and drug treatment groups. However, the absence of Ki67 protein in MCF-7 breast multicellular tumor spheroids is possibly related to the cells in G0 or S phase. These chemotherapeutic agents may affect the presence of ECM proteins in this in vitro model of micrometastasis of spheroids. These findings suggest that the possible mechanism of cell death in Doxorubicin and Docetaxel/Doxorubicin treatment groups is related to apoptosis through the p53 pathway. However, we considered the possiblity that there is another control mechanism for the Docetaxel group.Item The effect of oestradiol and neta on immunohistochemical staining of iNOS and eNOS in coronary arteries of ovariectomized rats(2006) Koyuncu F.M.; Ozbilgin K.; Kuscu N.K.; Inan S.; Vatansever S.; Ceylan E.Aim: The postmenopausal period is associated with increased risk for coronary atherosclerosis, and the effect of hormone replacement therapy in reducing this risk is controversial. Previous studies reported that nitric oxide synthetase (NOS) level might be important for the development of atherosclerosis, but no study has shown the interaction between hormone replacement therapy and endothelial NOS and inducible NOS intensity on coronary arteries yet. Our goal was to find out the immunostaining intensity of endothelial NOS and inducible NOS in ovariectomized rats which received oestradiol and norethisterone treatment. Methods: We performed bilateral ovariectomy in 15, female, 90-day-old Wistar rats with an average weight of 250 grams. After waiting for 4 weeks for the menopausal state, they were divided into 3 groups to receive either placebo, 0.1 mg/day 17-β-oestradiol (group E2), or 0.1 mg/day 17-β-oestradiol + 0.1 mg/day norethisterone acetate (group E2-NETA) for 5 weeks. Another group included 5, normal, adult, female intact rats and served as controls. At the end of the treatment, all rats were sacrificed and coronary arteries were stained with inducible NOS and endothelial NOS polyclonal antibodies using streptavidin-biotin technique. Results: The immunostaining of inducible NOS was prominent in perivascular connective tissue of the ovariectomized group but not in the control group. The inducible NOS immunostaining immunoreactivity was not detected in either treated groups. Immunostaining intensity of endothelial NOS did not differ in any 4 groups with similar staining. Conclusion: The present findings indicate that hormone replacement therapy down-regulates iNOS expression in coronary arteries of ovariectomized rats, and reduced iNOS may likely be involved in estrogen's beneficial effects.