Intraoperative evaluation of testicular vascularization and perfusion in rat testicles with indocyanine green (ICG)/near-infrared (NIR) fluorescent imaging after torsion-detorsion and reperfusion.

INTRODUCTION: 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 4th 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.

Role of Proopiomelanocortin in Preventing Miscarriage.

Objective: To compare the distribution of proopiomelanocortin (POMC) in decidua and placenta samples from missed abortion and voluntary termination cases in order to research the effects in the etiology of missed abortion. Study Design: Decidual materials were collected from patients who were diagnosed with missed abortion (n=19) and legal voluntary termination cases (n=15) under 10 gestational weeks. Materials were divided into 2 groups for examination. For all samples, POMC primary antibody was performed by immunohistochemical staining. The number of stained cells was calculated by using the H-score technique. Results: In the missed abortion group the mean age was 28.7 (18­41), and in the control group the mean age was 27.5 (21­37). POMC immunoreactivity was determined to be lower in the parenchyma and placenta of the missed abortion group than those of the control group. POMC immunoreactivities were found to be higher in both the syncytiotrophoblast and cytotrophoblast cells of the missed abortion group than those of the control group (p<0.005). Conclusion: POMC has become a paradigmatic polypeptide precursor and has a role in the parturition process. Local production of POMC in placenta and decidua may influence pregnancy and may have a role in missed abortion pathogenesis.

The role of selectins in the first trimester pregnancy loss.

OBJECTIVE: 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 = 0.007, p = 0.007). 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 < 0.007, 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.

Effect of raloxifene and atorvastatin in atherosclerotic process in ovariectomized rats.

AIM: 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-α). 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-α 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-α on endothelial cells. Combined treatment with these drugs shortly after menopause might play a potential preventive role in the early stages of atherosclerosis development.

Regulatory-T, T-helper 1, and T-helper 2 cell differentiation in nasal mucosa of allergic rhinitis with olive pollen sensitivity.

BACKGROUND: Allergic rhinitis (AR) is a disease in which T-helper (Th)2 response is predominant and its pathogenic mechanism is still poorly understood. AIM: To evaluate the possible role of Th1, Th2 and regulatory-T (Treg) cells in the pathogenesis of AR. METHODS: This case-control study enrolled 41 patients with seasonal AR (10-62 years old), sensitive to olive pollens, and 15 healthy controls (18-60 years old). Nasal biopsy was performed and specimens of nasal lavage fluid were obtained from all participants. The levels of interleukin (IL)-4, IL-10, interferon (IFN)-γ and transforming growth factor-ß (TGF-ß) were measured in nasal lavage fluid specimens. The expression of FOXP3, GATA-3 and T-bet was measured by immunohistochemical methods in the nasal biopsy specimens. RESULTS: The levels of IFN-γ in the group with AR were significantly lower than those in the control group (p = 0.008). The levels of IL-4, IL-10 and TGF-ß did not differ between the two groups. The expression of FOXP3 and T-bet in patients with AR was significantly lower than that in the control group (both p = 0.001). Expression of GATA-3 in the nasal mucosa was similar between the groups (p = 0.2). The ratios of T-bet/GATA-3 and FOXP3/GATA-3 in the AR group were significantly lower than those in the control group (p = 0.001). CONCLUSION: Insufficient Treg and Th1 cells may be associated with the allergic inflammation that may be attributed to the Th2 immune response in patients suffering from AR who are sensitive to olive pollen.

Immunohistochemical determination of mTOR pathway molecules in ovaries and uterus in rat estrous cycle stages.

mTOR is a member of the PI3K/Akt/mTOR signaling pathway that participates in cell growth, proliferation, protein synthesis, transcription, angiogenesis, apoptosis and autophagy. mTOR and MAPK pahways are two important key signal pathways which are related to each other. We investigated the role of mTOR and other signaling molecules in rat ovaries and uteruses in stages of the estrous cycle. Young adult female rats were divided into four groups as proestrous, estrous, metestrous and diestrous according to vaginal smears. Immunohistochemical staining of mTORC1, IGF1, PI3K, pAKT1/2/3, ERK1 and pERK1/2 was performed and pAKT1/2/3 and ERK1 were also analyzed using western blotting on ovarian and uterine tissue samples. According to our results, PI3K/Akt/mTOR and ERK/pERK showed an increase in the rat ovulation period. When all the groups were evaluated the immunoreactivities for all of the antibodies were detected in the oocytes, granulosa and theca cells, corpus luteum and stroma of ovary and lamina propria, surface and glandular epithelium of uterus with the strongest observed with anti-ERK1 antibody and then with a decreasing trend with anti-mTORC1, anti-pAkt1/2/3, anti-IGF1, anti-PI3K and anti-pERK1/2 antibodies in the proestrus and estrus stages. Differently from other parts of the ovary, highest antibody expression in the corpus luteum was observed in the metestrous stage. Moreover, the existence of pAKT1/2/3 and ERK1 proteins was confirmed with the Western blotting technique. We suggest that mTOR and mTOR-related ERK signaling molecules may participate in the rat ovulation process.

Prenatal administration of granulocyte-macrophage colony-stimulating factor increases mesenchymal vascular endothelial growth factor expression and maturation in fetal rat lung.

The aim of this study was to determine influence of prenatal granulocyte-macrophage colony-stimulating factor (GM-CSF) administration on lung growth, maturation, and vascular endothelial growth factor (VEGF) expression. Twenty Wistar rats received sterile saline (1 mL) or recombinant human GM-CSF (50 micro g/kg) on day 16 of pregnancy. Rats were sacrificed on days 18 and 20 of gestation. H-score for VEGF was calculated immunohistochemically. Alveolar VEGF expression on days 18 and 20 of gestation was significantly higher in the GM-CSF group (P < .01). Increase in VEGF with prenatal GM-CSF administration indicates that GM-CSF may stimulate lung growth and maturation and may be protective against lung disease due to prematurity.

Antenatal administration of granulocyte-macrophage colony-stimulating factor increases fetal lung maturation and endothelial nitric oxide synthase expression in the fetal rat lung.

OBJECTIVES: We investigated how maternal administration of granulocyte-macrophage colony-stimulating factor (GM-CSF) induced fetal lung maturation compared with dexamethasone and whether maternal administration of GM-CSF and dexamethasone influenced the fetal lung eNOS expression. STUDY DESIGN: Thirty pregnant rats were divided into three groups of 10 rats each to receive GM-CSF, dexamethasone or saline solution at 16 days of gestation. Lung maturation using bronchial area and immunohistochemical lung airway epithelium and the vascular endothelial eNOS expression, using H Scores, were evaluated at 18 and 20 days of gestation. The statistical analysis was done with the Kruskal-Wallis test for comparisons of more than two groups and the Mann-Whitney U-test as a post hoc test using SPSS for windows release 10.0. Values of p>0, 0.05 were considered significant. RESULTS: On the 20th day of gestation both GM-CSF and dexamethasone injections caused a significant increase in fetal lung bronchial area, as compared with the controls (24.9%, 36.8%, 13.4%, respectively, p=0.001). eNOS immunoreactivity was observed in the endothelium of large pulmonary vessels and large and small airway epithelium on the 18th and 20th day of gestation. Maternal GM-CSF and dexamethasone increased lung eNOS expression in the airway epithelium when compared to controls. CONCLUSION: Maternal administration of GM-CSF induced fetal lung maturation and this effect may be mediated, at least partly, by an increase in the eNOS expression.

A comparative study of the effect of raloxifene and gosereline on uterine leiomyoma volume changes and estrogen receptor, progesterone receptor, bcl-2 and p53 expression immunohistochemically in premenopausal women.

OBJECTIVE: To compare the mechanism of action of raloxifene and gosereline induced shrinkage of leiomyomas via estrogen receptor, progesterone receptor, bcl-2 and p53 expression immunohistochemically. STUDY DESIGN: Thirty-two premenopausal women affected by uterine leiomyomas were randomized into two equal groups. Group A was treated with gosereline (3.6 mg subcutaneous injection monthly) and group B was treated with raloxifene (60 mg daily per os) for 3 months before undergoing surgery. At entry and at the end of the treatment the leiomyoma volume was measured ultrasonografically and the volume change was calculated. Immunohistochemical detection of estrogen receptor (ER), progesterone receptor (PR), bcl-2 and p53 were performed on leiomyoma tissue samples from group A, group B and the matched-control group. H-scores for ER, PR, bcl-2 and p53 were calculated. The mean volume changes of leiomyomas and immunohistochemical H-score differences of ER, PR, bcl-2 and p53 were compared between groups. RESULTS: The leiomyoma volume decreased significantly after treatment in gosereline group from baseline of 65 cm(3) to 35 cm(3), and in raloxifene group from 68 cm(3) to 50 cm(3), p<0.05. The difference between the before and after treatment leiomyoma volumes between the two treatments was not statistically significant. H-score of ER expression was significantly lower in gosereline group compared to control group (54.4 versus 113.2, p = 0.001), whereas H-score of PR expression was significantly lower with both gosereline and raloxifene groups compared to control group (64.8 for gosereline versus 94.6 for control, 73.6 for raloxifene versus 94.6 for control, p = 0.001). The bcl-2 expression was higher in both gosereline and raloxifene groups compared to control group (173.7 for gosereline versus 94.7 for control, 179.7 for raloxifene versus 94.7 for control, p = 0.001). The p53 expression was only lower with gosereline than the control group (169.4 versus 205.6, p = 0.001), whereas there was no significant change between the raloxifene group and the control group (201.9 versus 205.6) (p>0.05). CONCLUSION: Raloxifene was as effective as gosereline in reducing leiomyoma volumes. Decreased PR expression may be a mechanism for tumor growth reduction in raloxifene treatment. In both treatment modalities, the mechanism of shrinkage of leiomyomas could not be increased apoptosis mediated by bcl-2 and p53 expression and should be investigated by further studies.

The effects of pirfenidone on T helper cells in prevention of intraperitoneal adhesions.

Abdominal surgery is linked with peritoneal adhesions. We investigated that the anti-fibrotic agent pirfenidone (PFD) has immune modulating activities and evaluated its effects on the function of T helper type 1 (Th1), Th2 and T regulatory (Treg) cells, which may play important roles in peritoneal adhesions. Eighteen female Wistar rats underwent right-sided parietal peritoneal and right uterine horn adhesion model. Rats were randomized into 3 groups as group 1 (control) (closure of midline abdominal incision without any agent administrations), group 2 (closure of incision after intraperitoneal administration of PFD) and group 3 (closure of incision and only oral administration of PFD for 14 days). Relaparotomy was performed 14 days after the first surgery. Effect of PFD on adhesion formation was assessed on Th1, Th2 and Treg cells counts using Anti-T-bet, Anti-GATA-3 Anti-FOXP3 antibodies respectively. Th1 counts were moderate in the control group, and didn't show a significant difference between all groups. Th2 cell counts were very high in the control group, but both intraperitoneal and oral administration of PFD resulted in a significant reduction in Th2 cell counts. Treg cell counts were low in number in the control group. In the intraperitoneal administration of PFD group, Treg cell counts were significantly lower than control group. There was no difference of the Treg cells between control groups and the oral administration of PFD group. PFD has prevention effect on intraperitoneal adhesions. This prevention effect seems to be related with the reduction in the numbers of Th2 and Treg cells.

Effect of Pirfenidone on Vascular Proliferation, Inflammation and Fibrosis in an Abdominal Adhesion Rat Model.

AIM: To study the efficacy of pirfenidone for prevention of postoperative adhesion formation in an adhesion rat model. MATERIALS AND METHODS: Eighteen female Wistar rats were subjected to right-sided parietal peritoneum and right uterine horn adhesion model. Rats were randomized into three groups: group 1 (control) (closure of midline abdominal incision without any agent administration), group 2 (closure of incision after intraperitoneal administration of pirfenidone), and group 3 (closure of incision and only oral administration of pirfenidone for 14 days). Relaparotomy was performed 14 days after the first surgery. Effect of pirfenidone on adhesion formation was assessed on light microscopy by scoring vascular proliferation, inflammation, fibrosis, and collagen formation in the scarred tissue. Effect of pirfenidone on inflammation was assessed by measurement of transforming growth factor-ß and interleukin-17 levels in scarred tissue. RESULTS: The degree of vascular proliferation (1.32 ± 0.39 versus 2.34 ± 0.46, p < 0.001), inflammation (1.60 ± 0.70 versus 2.60 ± 0.52, p < 0.01), and fibrosis (1.50 ± 0.53 versus 2.40 ± 0.52, p < 0.01) were less prominent in group 2 compared to group 1, respectively. Only vascular proliferation was found to be less prominent in group 3 compared to group 1 (1.60 ± 0.42 versus 2.34 ± 0.46, p < 0.01). Intraperitoneal and oral administration of pirfenidone reduced tissue levels of inflammatory markers (TGF-ß and IL-17) in parietal and visceral peritoneum compared to control group. Intraperitoneal administration of pirfenidone compared to oral administration was more effective in reducing tissue levels of inflammatory markers. CONCLUSION: Pirfenidone is an effective agent on the prevention of postoperative vascular proliferation, inflammation and fibrosis in scarred tissue particularly with intraperitoneal administration.

Histopathological and ultrastructural effects of Losartan on embryonic rat kidney.

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-beta (TGF-beta 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-beta'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.

Distribution of furin, TNF-α, and TGF-ß2 in the endometrium of missed abortion and voluntary first trimester termination cases.

OBJECTIVE: To identify the role of furin, TNF-α, and TGF-ß2 in human missed abortion pathogenesis. STUDY DESIGN: Decidual materials were collected from patients diagnosed with a missed abortion (n = 10) (missed abortion group) and from legal voluntary termination cases at < 10 gestational weeks (n = 10) (normal pregnancy group). Tissue samples were collected from each group by dilation and curettage under mask anesthesia. For all tissue samples,furin, TNF-α, and TGF-ß2 primary antibodies were performed by immunohistochemical staining. The number of stained cells was evaluated by using the H-score technique. RESULTS: In immunohistochemical examination, the immunoreactivities of furin, TNF-α, and TGF-ß2 were found to be higher in syncytiotrophoblastic cells in the missed abortion group than in the normal pregnancy group (p < 0.005). Additionally, high immunoreactivity of TNF-α and TGF-ß2 molecules was established only in cytotrophoblastic cells of missed abortions (p < 0.005) in examination at decidual cells of the missed abortion group; furin immunoreactivities were detected higher in the missed abortion group than in the control group, but TNF-α and TGF-ß2 immunoreactivity were increased in number in the normal pregnancy group (p < 0.005). CONCLUSION: It is considered that high levels offurin and the 2 furin-related proteins (TNF-α and TGF-ß2), which play important roles in proliferation, invasion, migration, differentiation, and survival of cells, may be the reason of proceeding decidualization, placentation, and prevention from abortion, in spite of terminating thefetal life.

The higher heparin-binding epidermal growth factor (HB-EGF) in missed abortion.

OBJECTIVE: Heparin-binding epidermal growth factor (HB-EGF) has pleiotropic biological functions in the female reproductive tract. HB-EGF has a function in the menstruation cycle, implantation, decidualization, placenta development, and also inhibition of apoptosis. This study aims to investigate a possible role of HB-EGF in missed abortion. MATERIALS AND METHODS: Decidual and placental tissue samples were obtained from women with unwanted pregnancy as the control group and from women with missed abortions as the patient group. Immunohistochemistry was utilized to compare HB-EGF expression of fibroblast and decidual cells in uterine decidual stroma and fibroblasts and mesenchymal cells in placental villous stroma; the TUNEL technique was used to detect apoptotic cells within the decidual and placental tissues of the two groups. RESULTS: It was demonstrated that HB-EGF expression in both uterine decidual stroma and placenta stroma was increased in the missed abortion group (142.70 ± 12.80; 116.10 ± 14.16, respectively), compared with the normal pregnancy group (101.60 ± 14.18; 81.60 ± 10.74, respectively). It was also shown that there was no difference in TUNEL (Terminal deoxynucleotide transferase dUTP Nick End Labelling) positive cells between the uterine decidual stroma (11.4 ± 3%; 13.6 ± 3%, respectively), placental villous stroma (13.7 ± 3%; 15.9 ± 3%, respectively), and cytotrophoblast-syncytiotrophoblast cells (7.3 ± 2; 9.8 ± 3, respectively) of the two groups. CONCLUSION: This data supports the hypothesis that increased HB-EGF expression in a missed abortion may prevent the discharge of the dead fetus.

Increased expression of angiogenic markers in patients with seasonal allergic rhinitis.

BACKGROUND: Increased vascularity due to neo-angiogenesis is an essential part of airway remodelling. Vascular endothelial growth factor (VEGF), CD34 and von Willebrand's factor (FvW) are known angiogenic markers. Angiogenesis and airway remodelling has been documented in asthma but not in allergic rhinitis. OBJECTIVE: We aimed to investigate the presence of increased angiogenesis and its relation to angiogenic molecules, namely VEGF, CD34 and FvW, in endothelial cells of nasal mucosa in patients with seasonal allergic rhinitis (SAR), using three different immunohistochemical analysis methods, namely HSCORE, microvessel density (MVD) and vascular surface density (VSD). The findings in allergic rhinitis were compared with the findings in nasal septal deviation (NSD), which is not associated with increased angiogenesis. METHODS: Twenty patients with symptomatic SAR, who were not under treatment, were enrolled in the study. Ten patients with NSD, who needed surgical therapy, served as the control group. Demographic characteristics did not differ between the two groups. Inferior turbinate biopsy was obtained from SAR patients and control patients, under local anaesthesia and during surgery respectively. All biopsies were evaluated for angiogenesis on the basis of VEGF, CD34 and FvW by two blinded histologists using three immunohistochemical analysis methods (HSCORE, MVD and VSD).Results. HSCORE, estimated on the basis of each staining technique, showed statistically significant differences among the two groups (p=0.002; p=0.045; p=0.016, respectively). Anti-CD34 and anti-VEGF showed higher MVD values in SAR when compared to the controls (p=0.038; p=0,009, respectively). No statistically significant difference was found in Anti-FvW-based MVD between SAR patients and controls (p=0.071). The measurements of VSD for FvW and VEGF from nasal biopsy specimens displayed a statistically significant difference between the two groups (p=0.004; p=0.0001, respectively). However, measurement of VSD for CD-34 was not significantly different between the groups (p=0.086). On the other hand, morphometric data obtained by all three methods did not correlated. CONCLUSION: There are a few studies that have investigated the essential role of angiogenesis in the pathogenesis of allergic rhinitis. We conclude that, increased angiogenesis may be as prominent in patients with allergic rhinitis as in patients with non-allergic nasal pathologies and may play an important role in the remodelling of nasal mucosa of subjects with SAR.

The effect of tibolone on endometrial IGF-1 and IGFBP-1 levels in ovariectomized rats.

OBJECTIVE: The goal of this study was to search the effects of two different doses of tibolone on endometrial IGF-1 and IGFBP-1 levels in ovariectomized rats. METHODS: Eighteen adult, female, 80-90-days-old, Wistar rats with an average weight of 250 g underwent bilateral ovariectomy under general anesthesia. After waiting for 4 weeks, they were randomized into three groups to receive either oral tibolone in two different doses or placebo. The treatment was continued for 5 weeks, and then the rats were sacrificed and the endometria were analyzed. RESULTS: Low columnar epithelium of the endometrial surface, longer epithelium and stratified squamous epithelium were seen in the control, low-dose and high-dose groups, respectively. The staining intensity of IGF-1 was mild in control, and moderate in both treatment groups, the difference between control the treatment groups was significant (P=0.015 for group L, and P=0.03 for group H). The staining intensity of IGFBP-1 was moderate in control, and strong in groups L and H. Again the difference was significant between control and both treatment groups (P=0.039 for grup L, and P=0.03 for group H). No significant difference was noted between each treatment group for both IGF-1 and IGFBP-1. CONCLUSION: Tibolone caused histological changes in endometrium and stimulated IGF-1 and IGFBP-1 staining. Both low and high dose treatments led to moderate and strong staining intensities for IGF-1 and IGFBP-1, respectively. The strong staining intensity of IGFBP-1 is likely due to the progestagenic effect of tibolone.

A histological analysis of the protective effect of ischemic preconditioning in the rat retina.

PURPOSE: Ischemic preconditioning (IP) protects the retina from the damaging effect of subsequent ischemia in vivo. We aimed to investigate the histological alterations induced by the protective effect of IP to the retina. METHODS: The eyes of the rats were rendered ischemic by intra-ocular pressure (IOP) elevation. IP procedure consisted of producing ischemia for 5 minutes. Sham operation was similar to IP procedure except the pressure elevation. The operational eyes of sham and IP group underwent 60 minutes of ischemia 24 hours after the first procedure. The eyes contralateral to the experimental eyes made up the control group. The eyes were histologically analysed one week after the ischemia. RESULTS: The total retinal thickness of the sham group was significantly less than total retinal thickness of the control group (p < 0.001). There was not a significant difference between control and IP group regarding the total retinal thickness (p > 0.05). The thickness of the inner retinal layers of the sham group were significantly less than corresponding retinal layers of the control group (p < 0.001). The inner plexiform layer (IPL) and inner nuclear layer (INL) thickness values of the sham group were significantly less than same layers of the IP group (p < 0.001). Ganglion cell layer (GCL) thickness of the IP group was significantly less than GCL thickness of the control group (p < 0.001). IPL thickness of the IP group was significantly less than that of control group's (p < 0.05). The GCL and total retinal thickness of the IP group were significantly more than thickness of the corresponding layers of the sham group (p < 0.05). CONCLUSION: IP considerably protects inner retinal layers from subsequent ischemic damage in a high IOP ischemic model. This endogenous process could further be utilized to tailor specific neuroprotective strategies for retinal cells.

Immunolocalization of integrins and fibronectin in tubal pregnancy.

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 (alpha 3, alpha V, beta 1, and alpha 2 beta 1) 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 alpha 3, beta 1, alpha V, and alpha 2 beta 1 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 alpha 3 and beta 1 integrins and fibronectin were increased in the normal part of fallopian tubes with ectopic pregnancies. Staining of beta 1 integrin was more intense than staining of alpha 3 and fibronectin, whereas there was no difference in alpha V and alpha 2 beta 1 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 alpha 3 and beta 1 integrins and fibronectin was strong in decidual cells, supporting tissue and placental villi, whereas alpha V and alpha 2 beta 1 staining was mild. We concluded that integrins, especially beta 1 and alpha 3, 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.

Protective effect of leukotriene receptor antagonist montelukast on smoking-induced lung injury in Wistar rats.

Increased activation of alveolar macrophage, neutrophil and mast cell has been proven in cigarette smoking (CS)-related lung disorders (CSLD). An increased production of cysteinyl-leukotrienes (LTs), which are mediators secreted from the mentioned cells, in response to CS has been shown in humans. The protective effect of LT1 receptor-1 antagonist (LTR-1AT) on CSLD is, however, not known. In this study we aimed to determine whether there is any protective effect of a LTR-1AT, montelukast (MK), on CSLD in Wistar rats. Nine controls and twenty-three smoke-exposed rats were enrolled into this study. Controls were exposed to non-filtered air, and the smoke-exposed rats were exposed to CS for 6 h/day, 6 days/week for three weeks. The CS-exposed rats were also treated with 0.1 mg/kg/day of MK or saline. Morphometric criteria for lung injury were determined as the mean linear intercept of alveolar septa (Lm), the volume density of alveolar septa (Vvspt) and the density of the alveolar surface area per unit volume of lung parenchyma (Sva.pa). Lung mast cells (LMC), which are a major source of LTs, were also counted. Results showed that Lm of the control group was significantly lower and Vvspt, Sva.pa of the controls were significantly higher compared to those of the CS-exposed groups. Animals treated with MK had significant protection against CSLD. Lm was significantly higher and Vvspt, Sva.pa were lower in the saline group than in the MK-treated group. The number of LMC in the CS-exposed groups was also significantly higher than that in the control group. Based on these results, one can suggest that some part of the pathogenesis of CSLD may be related to an enhanced LTs synthesis and LTR-1AT. Therefore, montelukast may protect against active or passive smoking-induced lung injury and related disorders.

TGF-ßs and Smads activities at the site of failed neural tube in the human embryos.

AIM: Transforming growth factor ß (TGF-ß) and Smads control intracellular signaling pathways in neurulation. Although previously reported similar experimental animal studies, the aim of this human study is to investigate the expression of TGF-ß (1,2,3) and Smads (1,2,3,6,7) in aborted human fetuses with myeloschisis. MATERIAL AND METHODS: Twelve human fetuses with neural tube defect were obtained. They were stained with antibodies against TGF-ß1, TGF-ß2, TGF-ß3, Smad (1,2,3), Smad 6 and Smad 7 using the indirect immunohistochemical technique. RESULTS: We noted mild immune reactivity of TGF-ß1 and TGF-ß2 in the open neural plate, motor neurons and surrounding tissue. Strong immune reactivity of TGF-ß3 was shown in only open neural plate and surrounding tissue. Immunoreactivity of all Smads noted negative except Smad7. CONCLUSION: These results suggested at the site where the neural tube failed to close, TGF-ß 1,2 and Smads 1,2,3,6 do not continue their activity and decrease with internal timing of embryonic development. Additionally ectodermal layers are considered by embryo as "not closed wound" and TGF-ß3 activity may be an effort to repair the failed closure.