Investigation of functional sperm membrane integrity after consequence of anti-hypoosmotic swelling test technique: an electron microscopic study.

OBJECTIVE: Infertility is described as the inability of a couple to conceive after 1 year of unprotected intercourse. Male factor is assumed to be responsible for 50% of cases of infertile couples. It is a common clinical problem, affecting approximately 13-15% of couples worldwide. In this study we aimed to investigate the effects of hypoosmotic swelling (HOS) and anti-HOS tests on sperm membrane integrity at the ultrastructural level. STUDY DESIGN: Twenty-nine infertile and 10 fertile men were included in this study. The fertile and infertile subjects were classified according to Kruger and WHO criterion. All semen samples were examined and ana-yzed based on WHO guidelines. Sperm viability was determined by using the eosin Y staining method. After HOS and anti-HOS tests wore applied, the samples were evaluated at the ultrastructural level. RESULTS: Normal structural features of all regions of sperm were observed in sections of sham normospermia. Some histopathological changes wore seen in HOS and anti-HOS group sections. CONCLUSION: The HOS procedure was found not to cause degenerative changes in the sperm ultrastructure. The anti-HOS procedure can be applied in normospermic and oligospermic groups.

Protective effects of ethyl pyruvate in cisplatin-induced nephrotoxicity.

This study was performed to investigate the effect of ethyl pyruvate on changes in renal functions and oxidative stress related renal injury caused by cisplatin (cis-dichlorodiammine platinum-II; CDDP). Male Wistar albino rats were divided into four groups (n = 8): (1) control group (1 ml Ringer's lactate solution i.p.); (2) ethyl pyruvate (EP) group (50 mg/kg Ringer's EP solution (REPS) i.p.); (3) cisplatin group (a single dose of cisplatin (5 mg/kg, i.p.); and (4) cisplatin + EP group (a single dose of cisplatin (5 mg/kg, i.p.) + REPS 50 mg/kg/day, i.p.) for five days. At the sixth day, kidneys of rats were mounted to a Langendorff apparatus. Renal perfusion pressures were recorded. Blood samples were taken for serum urea, creatinine, total oxidant status (TOS), total antioxidant status (TAS) and oxidative stres index (OSI) evaluations. Kidney tissues were obtained for malondialdehyde (MDA) analyses and histopathological examination. Perfusion pressures, serum urea, creatinine, TOS, OSI and tissue MDA levels were found significantly higher, whereas TAS was notably lower in cisplatin group. Histopathological examination showed apparent renal paranchymal injury in cisplatin group. In cisplatin + REPS group, perfusion pressures, serum urea, creatinine and tissue MDA levels were decreased. Moreover, EP co-administration provided less inflammatory cell infiltration, tubular dilatation, whereas TOS, TAS and OSI improved significantly versus cisplatin group. These findings show that EP has protective effects against cisplatin nephrotoxicity.

Effects of hyperbaric oxygen on crystalline lens and retina in nicotine-exposed rats.

OBJECTIVE: To determine histopathological changes on crystalline lens and retina of rats after subcutaneous injection of nicotine and to examine the effects of hyperbaric oxygen (HBO) on these changes related to nicotine exposure. METHODS: Twenty-eight female Sprague-Dawley rats were enrolled in the study and the rats were divided into four equal sized groups randomly (Group N: the rats exposed only to nicotine, group HB: the rats received only HBO, group N+HB: the rats that underwent to nicotine injection and subsequently received HBO, group C: the control group that neither exposed to nicotine nor received HBO). The rats were sacrificed by decapitation method and all were enucleated immediately after scarification. Tissue samples from crystalline lens, lens capsule, and the retina from the right eyes of the rats were examined by light microscopy. RESULTS: While the histological appearances of the retina and the lens was similar in group HB, group N+HB, and the control group; group N showed some pathological changes like decrement in the retinal ganglion cell density, atrophy of the retinal nerve fiber layer, congestion of the vessels in the optic nerve head, thinning of the internal plexiform layer, thinning of the lens capsule, and transformation of the anterior subcapsular epithelium into squamous epithelia. DISCUSSION: Subcutaneous injection of nicotine was found to be related with some pathological changes in the retina and lens of the Sprague-Dawley rats. However HBO caused no significant negative effect. Furthermore, the histopathological changes related to nicotine exposure in the lens and retina of the rats recovered by the application of HBO.

Comparison of the anti-diabetic and nephroprotective activities of vitamin E, metformin, and Nigella sativa oil on kidney in experimental diabetic rats.

Objectives: In this study, we aimed to evaluate and compare the nephroprotective and possible anti-diabetic effects of vitamin E, metformin, and Nigella sativa. Materials and Methods: Thirty male Wistar Albino rats were randomly divided into control, experimental diabetes (DM), vitamin E + DM, Metformin + DM, and N. sativa + DM. For experimental diabetes induction, IP 45 mg/kg streptozotocin was administered. Rats in vitamin E + DM, Metformin + DM, and N. sativa + DM received 100 mg/kg vitamin E, 100 mg/kg metformin, and 2.5 ml/kg N. sativa oil for 56 days. After the experiment, all animals were sacrificed, and blood and kidney samples were collected. Results: The blood urea level of the DM group was significantly higher (P<0.01) than the control group. Urea levels in vitamin E, metformin, and N. sativa groups were similar to the control group (P>0.05) but significantly different from the DM group (P<0.01). Bax, caspase-3, and caspase-9 immunopositivity intensity were quite low in the control group, and similar to the N. sativa group (P>0.05). Bcl-2 immunopositivity density was highest in the N. sativa group, similar to the control group in terms of percentile area (P>0.05). Conclusion: When all three treatment methods were compared in terms of their effectiveness in alleviating DM and DN, it was determined that the most successful result was obtained with N. sativa oil.

Severe preeclampsia complicated by HELLP syndrome alters the structure of Hofbauer and syncytiotrophoblast cells: ultrastructural and immunohistochemical study.

Background: Hemolysis, elevated liver enzymes, low platelet count (HELLP) syndrome is generally considered to be a variant or complication of preeclampsia. It is a life-threatening obstetric complication. Objectives: To evaluate the immunohistochemistry and ultrastructural of syncytiotrophoblastand Hoffbauer cells in placental villi of patients with HELLP syndrome. Methods: Two groups of patients with a total of 50 full-term human placentas (n = 25 in each group) were assigned as the control (normotensive) and HELLP syndrome. Placental tissue samples were fixed in 10% neutral formalin and paraffin-embedding protocol was performed. We prepared 5 µm sections for histological and immunohistochemical staining. Sections were immunostained with Hoffbauer cell marker CD68. For transmission electron microscopy (TEM), placental tissue samples were fixed in 2.5% buffered glutaraldehyde and then, in 1% osmium tetroxide for routine ultrastructural examinations. Results: When the HELLP group fetal placental sections were examined, intracytoplasmic edema in syncytiotrophoblast, degenerative vacuoles, and degenerative findings on cell surface membranes were observed. Moreover, villous edema was remarkable. The number of CD68-positive Hoffbauer cells per villus control group sections was 0.23 ± 0.02 and the number of CD68-positive cells per villus in HELLP group placenta sections was 0.83 ± 0.12. The increase in the number of Hoffbauer cells per villus in the HELLP group was significant (P < 0.001). Compared with the control group, there was a significant increase in the number of Hoffbauer cells and syncytiotrophoblasts in the HELLP group, and degenerative changes were also observed in the ultrastructure of these cells. Conclusions: Pathology of the HELLP syndrome is in relation to CD68-positive placental macrophages.

The subconjunctival use of cetuximab and bevacizumab in inhibition of corneal angiogenesis.

PURPOSE: To investigate the effects of cetuximab and bevacizumab on experimental rat model of corneal angiogenesis. METHODS: The right eyes of 28 male Sprague-Dawley rats were included in silver nitrate cauterization-induced corneal angiogenesis model. They were divided into four groups: (1) silver nitrate cauterization-induced and 0.15 ml serum physiologic was given to the angiogenesis group, (2) bevacizumab was given 1.25 mg to the bevacizumab group, (3) cetuximab was given 5 mg to the cetuximab group, and (4) 1.25 mg bevacizumab plus 5 mg cetuximab were given to the bevacizumab + cetuximab group. All eyes were exposed to the treatment on days 1, 4, and 7 of the experiment, and drugs were given subconjunctivally. The left eyes were untreated and used as sham. On day 8, the treated eyes were evaluated biomicroscopically. Then, the rats were sacrificed, and corneal specimens were prepared for histopathologic examinations. RESULTS: The degree of angiogenesis inhibition was observed as 50.8% in bevacizumab, 54.3% in cetuximab, and 15.8% in bevacizumab + cetuximab groups by biomicroscopic evaluation. According to the histopathological and immunohistochemical findings obtained from the present study, the amount of angiogenesis was determined to have decreased considerably in both the bevacizumab and cetuximab groups; also, relatively less inhibiton was observed in the bevacizumab + cetuximab group. CONCLUSION: Subconjunctival injection of cetuximab and bevacizumab is effective in reducing corneal angiogenesis in silver nitrate cauterization induced angiogenesis model of rats. Further investigation is needed to assess the potential side-effects of the drugs, especially cetuximab.

Protective effects of necrostatin-1 on doxorubicin-induced cardiotoxicity in rat heart.

Background: Doxorubicin (Dox) is one of the most effective antineoplastic drugs which has severe cardiotoxic effects, limiting its clinical usage. Though the exact mechanism of doxorubicin-induced cardiotoxicity is yet to be elucidated, it is shown that production of reactive oxygen species (ROS) increases oxidative stress and leads to cardiomyocyte apoptosis and necroptosis which is also defined as a programmed cell death.Purpose: In the present study, we investigate the effects of necrostatin-1 (Nec-1)-an inhibitor of receptor interaction proteins 1 (RIP1) and necroptosis-on doxorubicin-induced cardiotoxicity in rats.Research Design: Hearts were isolated and perfused by the Langendorff system in all four groups. Perfusion pressure (PP), left ventricular developed pressure (LVDP) and heart rate per minute (HR), LV (dP/dt) max, and LV (dP/dt) min which shows cardiac contractility and relaxation were recorded.Results: Results showed that PP significantly increased with Dox treatment and significantly decreased with Nec-1 treatment, while HR, LVDP, LV (dP/dt) max, and LV (dP/dt) min values significantly decreased with the Dox-treated group and significantly increased with Nec-1 treatment. Also with Nec-1 treatment, gene expression levels of anti-apoptotic Bcl-2 significantly increased and pro-apoptotic protein Bax, apoptotic marker caspase-3, and Nox-2 significantly decreased compared to the Dox-treated group. In heart tissues, MDA levels were significantly increased with Dox and decreased with Nec-1 treatment. These results were supported by the histological analysis indicated that Nec-1 reduced doxorubicin-induced cellular injury.Conclusions: In conclusion, our data indicate that Nec-1 ameliorates doxorubicin-induced cardiotoxicity by reducing oxidative stress injury and attenuating apoptosis and necroptosis.

Effects of black cumin seed oil on oxidative stress and expression of membrane-cytoskeleton linker proteins, radixin, and moesin in streptozotocin-induced diabetic rat liver.

Background and Aim: This study examined the effects of black cumin seed oil treatment on oxidative stress and the expression of radixin and moesin in the liver of experimental diabetic rats. Materials and Methods: Eighteen rats were divided into 3 equal groups (control, diabetes, treatment). The control group was not exposed to any experimental treatment. Streptozotocin was administered to the rats in the diabetes and treatment groups. A 2.5 mL/kg dose of black cumin seed oil was administered daily for 56 days to the treatment group. At the conclusion of the experiment, the blood level of malondialdehyde (MDA) and glutathione (GSH) was measured. The expression level and the cellular distribution of radixin and moesin in the liver were analyzed. Results: The plasma MDA (3.05±0.45 nmol/mL) and GSH (78.49±20.45 µmol/L) levels in the diabetes group were significantly different (p<0.01) from the levels observed in the control group (MDA: 1.09±0.31 nmol/mL, GSH: 277.29±17.02 µmol/L) and the treatment group (MDA: 1.40±0.53 nmol/mL, GSH: 132.22±11.81 µmol/L). Immunohistochemistry and western blotting analyses indicated that while the level of radixin was not significantly between the groups (p>0.05) and moesin expression was significantly downregulated (p<0.05) in the experimental group, the treatment was ineffective. Conclusion: The administered dose was sufficient to prevent oxidative stress, but was not sufficient to alleviate the effects of diabetes on moesin expression in hepatic sinusoidal cells.

Intracellular Ca(2+) levels in rat ventricle cells exposed to extremely low frequency magnetic field.

OBJECTIVE: Electromagnetic fields can affect intracellular Ca(2+) levels. The aim of this study was to determine the changes intracellular Ca(2+) concentration in cardiac ventricle cells of rats exposed to 0.25 mT (2.5 Gauss) magnetic field. METHODS: Forty-five male rats were introduced to this study. The rats were divided into three groups: control, sham, and experiment. The experimental group was exposed to 0.25 mT extremely low frequency (ELF) magnetic field for 14 days, 3 h/day. The sham group was treated like the experimental group, except for elf-magnetic field exposure. The control group was not subjected to anything and differed from the experimental group and sham group. In the end of experiment, rats were sacrificed, cardiac tissue was removed, and these were fixed in 10% neutral formalin. Then, ventricular cells were stained by Alizarin red staining method. RESULTS: In the light microscopic examinations of control groups, in myofibril structures between groups, changes were not observed. In myofibril regions of the experimental group compared to other groups, increased heterogen Ca(2+) accumulations were found. CONCLUSION: ELF magnetic fields are used in daily life. The results of this study show that intracellular Ca(2+) accumulation in cardiac ventricles can increase in rats exposed to ELF magnetic field.

Is there a role for antioxidants in prevention of pulmonary hypoplasia in nitrofen-induced rat model of congenital diaphragmatic hernia?

BACKGROUND/PURPOSE: Many studies suggest a role for antioxidants in the prevention of lung hypoplasia in nitrofen-induced rat models with congenital diaphragmatic hernia (CDH). This study investigates the oxidative status and the histological outcome of prenatal administration of vitamins E and C with synergistic effect, and effect of N-acetylcysteine (NAC) to improve lung maturation of nitrofen-induced rats. METHODS: CDH was induced by maternal administration of a single oral dose of nitrofen on day 9.5 of gestation, and the Sprague-Dawley rats were randomly divided into five groups: nitrofen (N), nitrofen + vitamin C (NC), nitrofen + vitamin E (NE), nitrofen + vitamin C + vitamin E (NCE) and nitrofen + NAC (NNAC). A control group in which only vehicle was administered was included. Cesarean section was performed on day 21. Body weight (BW) and total lung weight (LW) of all fetuses with CDH were recorded; lung histological evaluation was performed, and protein content of lungs, determination of thiobarbituric acid reactive substances, and the protein carbonyls in tissue samples were determined. RESULTS: A total of 133 rat fetuses with CDH were investigated. The body weight and the lung weight of fetuses of all groups that were exposed to nitrofen were significantly decreased than of the control group (P < 0.05). The animals exposed to nitrofen with different antioxidants showed increased protein levels in lung tissue. However, in the NCE and the NNAC groups, protein levels were significantly increased than in the others. Malondialdehyde levels significantly decreased in the NCE and the NNAC groups when compared with the NC and the NE groups. In addition, the NCE and NNAC groups decreased protein oxidation to control levels, and no significant difference was observed between control and these two antioxidants groups. The N, NC, NE and NNAC groups showed minimal improvement in lung histology; the NCE groups showed the most improvement in lung histology when compared with the other nitrofen plus antioxidant groups. CONCLUSION: Prenatal administration of NAC and vitamin E in combination with vitamin C represented the best effects to avoid oxidative damage and protein content of the lungs in rat pups with CDH at birth.

Trolox is more successful than allopurinol to reduce degenerative effects of testicular ischemia/reperfusion injury in rats.

INTRODUCTION: Reperfusion surgery following testicular ischemia is a reproductive health threatening status and may result with organ dysfunction in men. The high level of reactive oxygen species (ROS) and cease of blood flow to the testis are the most important reasons of this testicular injury. Until today, numerous experimental studies reported that antioxidants might be efficient to alleviate oxidative stress induced organ dysfunction. For this purpose, in this study, we have investigated the protective effects of xanthine oxidase (XO) inhibitor, allopurinol, and ROS scavenger, trolox, in a comparative perspective in testicular ischemia reperfusion injury subjected rats. MATERIALS AND METHODS: Twenty-eight adult male Sprague Dawley rats were divided into four groups of seven animals in each; control, ischemia/reperfusion (I/R), allopurinol and trolox. The rats in control group did not receive any application. Animals in I/R, allopurinol and trolox groups were subjected to 2 h testicular reperfusion injury following 5 h ischemia. Intraperitoneally (i.p.) 1 ml isotonic, 200 mg/kg allopurinol and 50 mg/kg trolox were administered to the animals in these groups 30 min prior reperfusion. At the end of experiment, all animals were sacrificed and blood serum malondialdehyde (MDA) levels were measured. Histological sections were obtained from the testis and stained with hematoxylin and eosin (H&E), proliferating cell nuclear antigen (PCNA) and cleaved caspase-3. Apoptotic index was evaluated with TUNEL Assay. RESULTS: Severe morphological degenerations, increased serum MDA, cleaved caspase-3 and TUNEL Assay positivity rate, but reduced PCNA positivity rate was observed in ischemia and reperfusion group. Morphological degenerations, MDA level, apoptotic index and PCNA positive cell rate were slightly alleviated in allopurinol administered animals compared with ischemia and reperfusion group. Protection with trolox was more successful and the results of the analysis were similar to the control group. DISCUSSION: Ischemia that leading to testicular torsion is a reproductive health affecting problem and current surgical treatment methods might be insufficient to recover testis. Various types of ROS generating mechanisms in cell are limiting protective potency of allopurinol, and cocktail administration of different ROS inhibitors might be more effective. However, our results indicate that free radical scavenger trolox might be a candidate drug to alleviate degenerative effects of testicular ischemia reperfusion injury. CONCLUSIONS: This is the first study that demonstrates antioxidant trolox was more successful than XO inhibitor allopurinol to protect testis against ischemia and reperfusion injury in rats.

Combining platelet rich fibrin with different bone graft materials: An experimental study on the histopathological and immunohistochemical aspects of bone healing.

AIM: The current study was based on the hypothesis that the use of PRF with bone graft materials might increase bone regeneration and focus on the histopathological and immunohistochemical aspects following application of PRF with autogenous graft, xenograft and B-TCP in a rabbit model. MATERIAL AND METHODS: This study was performed on the twenty-eight male New Zealand divided into four group. Two defects with a diameter 10 mm were opened in calvarium. After PRF preparation, right defects were evaluated as empty defect or graft group, and left defects were evaluated as PRF test group. All animals were sacrificed at the end of 8 weeks and specimens were examined histopathologically and immunohistochemically. RESULTS: The most superior histopathological results were obtained in the autograft group. The combination of ß-TCP-PRF could not provide superiority over the ß-TCP group. The immunohistochemical results showed that, in the PRF/BTCP group, the expression of osteopontin and osteonectin was relatively higher compared to the only-BTCP group. CONCLUSION: In terms of new bone formation, autograft combined with PRF yielded superior results but the combination of ß-TCP-PRF had no effect compared to the only-BTCP group. However, further experimental and clinical studies might be beneficial to clarify the exact mechanism and results of combining PRF with bone grafts on bone healing process.

The demonstration of changes in bulbar conjunctiva surface epithelium in the psoriatic patients treated with PUVA.

We investigated the effects of disease itself and PUVA treatment on surface epithelium of conjunctiva in psoriatic patients (PP) before PUVA and after PUVA therapy and in 32 healthy volunteers. Squamous metaplasia was detected in PP both before and after PUVA therapy. We concluded that PUVA treatment applied together with preventive measures, would lead to less severe ocular side effects.

An experimental Staphylococcus aureus meningitis model for investigating induced leptomeningeal and subpial inflammation in rats: a transmission electron microscopy study.

OBJECTIVE: To evaluate leptomeningeal and subpial inflammatory responses of experimental Staphylococcus aureus bacteriemia following intraperitoneal and intravenous applications and to compare the inflammatory reactions in different regions of central nervous system. MATERIAL AND METHODS: Forty anesthetized rats were divided into four groups equal in number. The rats in group-I were given 1 ml suspension of Staphylococcus aureus intraperitoneally. Group-II was the control group of group I; it was administrated 1 ml 0.9% NaCl in water intraperitoneally. The rats in group-III were given the same amount of bacteria intravenously. Group IV was the control group of the group-III; it was administrated 1 ml 0.9% NaCl solution intravenously. The rats were sacrificed on the 21st day. Inflammatory changes of different regions of the central nervous system were examined under transmission electron microscopy. Statistical analysis was done by using variance analysis, Bonferroni, Tamhane post hoc, Student's t and univariate tests. RESULTS: Thoracic and occipital regions were the most vulnerable zones. Increasing of collagen tissue was the most detected inflammatory change. CONCLUSION: This experimental model can be used for inducing subpial and leptomeningeal inflammations and it may be developed for investigations of pathogenesis of leptomeningitis during systemic infections.

Investigation of the toxic effects of ropivacaine corneal endothelium by the impression cytological method.

OBJECTIVE: To investigate the toxic effects of ropivacaine on corneal endothelium by using the impression cytological method. METHODS: The study was performed between October and December 2004 in Dicle University Hospital, Diyarbakir, Turkey. Twenty-four eyes from 12 rats were used for the research. They were divided into 4 groups, each containing 3 different ropivacaine concentrations and a control. Immediately after enucleation, the corneas were excised and the endothelium was exposed to unpreserved ropivacaine 0.01, 0.1, or 1% and balanced salt solutions (BSS) as a control (6 corneas/group) for 20 minutes. The specimens were obtained by impression cytology method and stained with periodic acid shift. Then, they were examined under light microscope. RESULTS: Blurring at cell membrane borders, vacuolization at cell cytoplasm, hydropic degeneration and increase in toxic granulation were observed in the 1% ropivacaine group. Cytoplasmic hydropic degeneration was determined in the 0.1% ropivacaine group. Cell structures were normal and almost identical to the control group in the 0.01% ropivacaine group. CONCLUSION: In this study, 2 major conclusions were determined. The impression cytology method can be used in examination of corneal endothelium, and exposure of rat corneal endothelium to 0.01% ropivacaine solutions in vitro appears to be non-toxic.

Effects of Hyperbaric Oxygen Treatment on Renal System.

INTRODUCTION: Hyperbaric oxygen (HBO) treatment is steadily increasing as a therapeutic modality for various types of diseases. Although good clinical outcomes were reported with HBO treatment for various diseases, the multisystemic effects of this modality are still unclear. This study aimed to investigate the renal effects of HBO experimentally. MATERIALS AND METHODS: Fourteen New Zealand White rabbits were divided into 2 groups randomly as the control group and the study group. The study group received HBO treatment for 28 days (100% oxygen at 2.5 atmospheres for 90 minutes daily) and the control group was used to obtain normal renal tissue of the animal genus. After the intervention period, venous blood samples were obtained, and renal tissue samples were harvested for comparisons. RESULTS: Normal histological morphology was determined with Masson trichrome staining and periodic acid-Schiff staining in the control group. Atrophic glomerular structures, vacuolated tubule cells, and degeneration were detected in the renal samples of the study group with Masson trichrome staining. Additionally, flattening was observed on the brush borders of the proximal tubules, and tubular dilatation was visualized with periodic acid-Schiff staining. The histopathologic disruption of renal morphology was verified with detection of significantly elevated kidney function laboratory biomarkers in the study group. CONCLUSIONS: Our findings suggests that HBO has adverse effects on renal glomerulus and proximal tubules. However, the functional effects of this alteration should be investigated with further studies.

The effects of valproic acid, vitamin E and folic acid on ribs of rat fetuses in the prenatal period.

The present study was undertaken to determine histopathologic differences in the ribs of Wistar-albino rat fetuses prenatally exposed to valproic acid (VPA), folic acid (FA) and vitamin E (Vit E), and to compare their differential developmental susceptibility and morphological association with skeletal anomalies. VPA (300 mg/ kg), FA (300mg/kg) and Vit E (250mg/kg) were administered to rats on each of gestation days (GD) 7-9. Fetuses were collected on GD 21 and their ribs were examined for malformations. The fetuses were divided into four groups: blind-trial group, VPA group (vpa), VPA and Vit E group (vpa+vit e), valproic and FA group (vpa+fa). In each group; drug procedure, surgical procedure and histological methods were performed. Later, weights and lengths of fetuses in each group were compared and analyzed by one-way Anova test. As a result, maLformations in fetuses were determined and photographed by Nikon SMZ-2 steromicroscopy, using 2 x objective. Administration of single doses of VPA (300 mg/kg) resulted in weight and length loss between blind-trial and vpa group. However, length and weight differences between the other groups were not significant. The objective of the present study is to analyze morphological and histopathologic changes which may occur in a high-risk experimental model after the administration of VPA. In addition, protective roles of the administration of FA and Vit E are assessed.

Effects of trypan blue on corneal endothelium and anterior lens capsule in albino wistar rats: An investigator-masked, controlled, two-period, experimental study.

BACKGROUND: The development of continuous curvilinear capsulorrhexis (CCC) has contributed significantly to the tolerability and effectiveness of cataract extraction and intraocular lens implantation. Staining of the anterior capsule has become a popular method of increasing visibility when performing CCC. OBJECTIVE: The aim of this study was to determine, using scanning electron microscopy (SEM) and transmission electron microscopy (TEM), the highest concentrations of trypan blue dye that would not cause long-term toxicity after injection into the anterior chamber of rat eyes. METHODS: The eyes of healthy female albino Wistar rats were used in this investigator-masked, controlled, 2-period, experimental study conducted over 12 weeks at the Dicle University Experimental Animal Laboratory, Diyarbakir, Turkey. The rats were randomly divided into 5 groups of 4 using a random-number table. Each rat was administered a 0.05-mL injection of trypan blue into the right eye in 1 of the following concentrations: 0.4%, 0.2%, 0.1%, 0.05%, or 0.025%. A 0.05-mL pH-balanced saline solution was injected into the left eye of each rat to act as a control. At 1 day after injection and 4 weeks after injection (early period), 1 rat from each concentration group was euthanized and their eyes were enucleated. At 12 weeks after injection (late period) the remaining 2 rats from each group were euthanized and their eyes were enucleated. Corneal endothelial cells and the anterior lens capsule of the enucleated eyes were analyzed using SEM and TEM, and the results were compared with those of the control group. In the TEM analysis, the primary end point was the histopathologic changes in the cellular organelles when compared with those in the control group. In the SEM analysis, the primary end point was changes in cell shape, intracellular junctions, and density of the microvilli when compared with those in the control group. RESULTS: Forty eyes from 20 albino Wistar rats (mean [SD] age, 8.2 [1.6] weeks; mean [SD] weight, 175.6 [16.5] g) were used in the study. Each group of rats received a different concentration of trypan blue in the right eye. In the early period, both the 0.4% and 0.2% concentrations were associated with the impairment of the hexagonal structure of corneal endothelial cells and intercellular junctions. Those concentrations were also associated with an increased occurrence of cellular vacuolation, cytoplasmic edema, extensive granulation of the endoplasmic reticulum, pyknotic nuclei, and mitochondrial degeneration. In the late period, these changes were observed as persisting in a decreasing manner. With the 0.1% and 0.05% concentrations, the density of microvilli decreased, nuclei appeared normal, granulation of the endoplasmic reticulum and Golgi apparatus was active, and minimal levels of mitochondrial degeneration were observed. CONCLUSIONS: In this small experimental study in rat eyes, trypan blue at concentrations >0.025% was associated with impaired morphology and structure of corneal endothelial cells after short-term exposure. This effect continued in a decreasing fashion after long-term exposure. No significant changes were noted in the control group or the group administered the 0.025% concentration.

Effects of intravitreal ropivacaine on retinal thickness and integrity in the guinea pig.

BACKGROUND: Retrobulbar anesthesia is widely used for ocular surgery.Ocular complications are possible when retrobulbar anesthesia is accidentally injected intravitreally. OBJECTIVE: The aim of this study was to determine the relative retinal toxicitiesof ropivacaine hydrochloride, a local anesthetic, using various concentrations in guinea pigs. METHODS: This randomized, investigator-masked, experimental study wasconducted at the Department of Anesthesiology, Dicle University, Diyarbakir, Turkey. The right eyes of 18 guinea pigs were assigned to 1 of 3 treatment groups: 1%, 0.75%, or 0.5% ropivacaine. The right eye of each animal was injected intravitreally with 0.1 mL of 1%, 0.75%, or 0.5% ropivacaine. The left eye of each animal was injected with a balanced saline solution (control). The guinea pigs were euthanized 7 days after injection, and the retinal structures were examined using light microscopy. The total thickness of each retina was measured using an ocular micrometer. RESULTS: No histologic abnormalities were observed in the control eyes.Retinal damage of most of the retinal section was seen in the eyes receiving study drug. The eyes injected with 0.5% ropivacaine had a generally intact appearance, with the exception of some atrophy and disorganization. Overall, the eyes injected with 1% ropivacaine had significantly more extensive retinal thinning compared with the eyes injected with 0.75% or 0.5% ropivacaine (both, P < 0.01). In the eyes injected with 0.75% or 1% ropivacaine, disorganization of the structure of the retinal layers and atrophy were noted on histopathology. The mean total thicknesses of the retina were significantly less in all ropivacaine-treated eyes compared with that in the controls (P < 0.001). CONCLUSIONS: In this small experimental study, ropivacaine had concentration-dependent toxic effects on guinea pig retinas.

Reconstruction techniques in renal injury.

OBJECTIVE: Kidney repair reconstruction techniques are controversial. The conventional technique is suturing, but this is usually with further loss of viable tissue as it promotes scaring. In this animal model, we investigated the parenchymal effect of different sutures and methods. METHODS: We carried out this study in the year 2000 in the Animal Laboratory of Dicle University, Diyarbakir, Turkey. We used 40 New Zealand white rabbits in this study, randomized into 5 groups. We separated the left kidney from Gerota's fascia, and performed standard lacerations with incisions 5 mm deep and 10 mm in length. We performed no reconstruction procedure in group 1, and used homeostatic collagen powder in group 2. We primarily sutured the lacerations with chromic gut (4/0) in group 3, and sutured the kidney in group 4 with polyglactin (4/0). We wrapped the kidney with a polyglactin mesh in group 5. We sacrificed 2 rabbits in each group postoperatively on day 2, 15, 45 and 90, and performed left nephrectomy for histological investigation, and assessed interstitial inflammation. RESULTS: While group 5 established the best results, the other 4 groups had similar intermediate results. The pseudocapsule was visible macroscopically in the polyglactin mesh group. We could see mononuclear cell infiltration, dilatation of tubules, atrophy of tubules, and interstitial fibrosis in all groups except group 5. CONCLUSION: In this animal model, we found that the most appropriate repair material for kidney surgery was polyglactin mesh.