The Relationship Between Ischemia Time and Mucous Secretion in Vaginal Reconstruction With the Jejunal Free Flap: An Experimental Study on the Rat Jejunum.

Jejunum flap for reconstruction of the vagina provides a durable, stable coverage; patent tube passage; and natural esthetic appearance. However, excessive mucous secretion is a major drawback of the technique.We have recently presented our cases in which strict 3-hour ischemia with lower mucus secretion was applied. However, a quantitative analysis of goblet cells of the jejunum subjected to ischemia and ischemia-reperfusion injury on an animal model has not been reported to support this argument.Because goblet cells are responsible for the production and the maintenance of the mucous blanket, we aimed to determine whether goblet cell numbers decrease after ischemia and ischemia-reperfusion injury.This study was conducted on 3 groups of 10 animals. We applied to the rat jejunum only ischemia in group 1, one hour of ischemia followed by reperfusion in group 2, and 2 hours of ischemia followed by reperfusion in group 3. Histological samples taken from the jejunum exposed to ischemia and ischemia-reperfusion injury were evaluated in terms of goblet cell numbers, inflammation, apoptotic bodies, and necrosis.Goblet cell numbers significantly decreased in the group of animals exposed to ischemia and exposed to ischemia-reperfusion injury. We think that mucus hypersecretion of the jejenum can be limited by applying a longer period of ischemia time during free flap transfer in vaginal reconstruction.

Inhibition of neutral sphingomyelinase decreases elevated levels of inducible nitric oxide synthase and apoptotic cell death in ocular hypertensive rats.

Endoplasmic reticulum (ER) stress and excessive nitric oxide production via induction of inducible nitric oxide synthase (NOS2) have been implicated in the pathogenesis of neuronal retinal cell death in ocular hypertension. Neutral sphingomyelinase (N-SMase)/ceramide pathway can regulate NOS2 expression, hence this study determined the role of selective neutral sphingomyelinase (N-SMase) inhibition on retinal NOS2 levels, ER stress, apoptosis and visual evoked potentials (VEPs) in a rat model of elevated intraocular pressure (EIOP). NOS2 expression and retinal protein nitration were significantly greater in EIOP and significantly decreased with N-SMase inhibition. A significant increase was observed in retinal ER stress markers pPERK, CHOP and GRP78 in EIOP, which were not significantly altered by N-SMase inhibition. Retinal TUNEL staining showed increased apoptosis in all EIOP groups; however N-SMase inhibition significantly decreased the percent of apoptotic cells in EIOP. Caspase-3, -8 and -9 activities were significantly increased in EIOP and returned to baseline levels following N-SMase inhibition. Latencies of all VEP components were significantly prolonged in EIOP and shortened following N-SMase inhibition. Data confirm the role of nitrative injury in EIOP and highlight the protective effect of N-SMase inhibition in EIOP via down-regulation of NOS2 levels and nitrative stress.

Suppressive effect of astaxanthin on retinal injury induced by elevated intraocular pressure.

The aim of this study was to clarify the possible protective effect of astaxanthin (ASX) on the retina in rats with elevated intraocular pressure (EIOP). Rats were randomly divided into two groups which received olive oil or 5mg/kg/day ASX for a period of 8 weeks. Elevated intraocular pressure was induced by unilaterally cauterizing three episcleral vessels and the unoperated eye served as control. At the end of the experimental period, neuroprotective effect of ASX was determined via electrophysiological measurements of visual evoked potentials (VEP) and rats were subsequently sacrificed to obtain enucleated globes which were divided into four groups including control, ASX treated, EIOP, EIOP+ASX treated. Retinoprotective properties of ASX were determined by evaluating retinal apoptosis, protein carbonyl levels and nitric oxide synthase-2 (NOS-2) expression. Latencies of all VEP components were significantly prolonged in EIOP and returned to control levels following ASX administration. When compared to controls, EIOP significantly increased retinal protein oxidation which returned to baseline levels in ASX treated EIOP group. NOS-2 expression determined by Western blot analysis and immunohistochemical staining was significantly greater in rats with EIOP compared to ASX and control groups. Retinal TUNEL staining showed apoptosis in all EIOP groups; however ASX treatment significantly decreased the percent of apoptotic cells when compared to non treated ocular hypertensive controls. The presented data confirm the role of oxidative injury in EIOP and highlight the protective effect of ASX in ocular hypertension.

Corneal protein nitration in experimental uveitis.

Increased expression of inducible nitric oxide synthase (NOS-2) in inflammatory diseases like uveitis suggests that it contributes to the observed pathological state. The aim of this study was to evaluate corneal expression of NOS-2 and corneal protein nitration in a rat model of uveitis. A single injection of intravitreal lipopolysaccharide was used to induce uveitis. Corneal proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and visualized by Coomassie blue staining. Expression of NOS-2 and nitrotyrosine (NO(2)Tyr) formation were determined via immunohistochemistry and Western blot analysis. Total nitrate/nitrite levels in the vitreous were measured by spectral analysis via the Griess reagent. Immunohistochemical analysis revealed increased corneal NOS-2 and NO(2)Tyr immunoreactivity in rats with uveitis compared with controls. NOS-2 and NO(2)Tyr immunoreactivity was observed in and around basal cells in the corneal epithelium. Western blot analysis of corneal lysates showed multiple nitrated protein bands in uveitic rats. Spectrophotometric measurement of total nitrate/nitrite levels in the vitreous affirmed significantly increased levels of nitric oxide generation in uveitis (126 +/-2.63 microM/mg protein) compared with controls (65 +/-6.57 microM/mg protein). The presented data suggests that extensive formation of protein nitration and reactive nitrogen species in the cornea contributes to tissue destruction in uveitis. Hence, selective inhibition of NOS-2 may prevent long-term complications and lead to an improvement in the management of uveitis.

Reduced E-cadherin and alpha-catenin expressions have no prognostic role in bladder carcinoma.

In various human cancers, dysfunction of the E-cadherin-catenin complex is associated with a decrease in cellular and tissue differentiation, and with higher invasive and metastatic potentials. The objective of this study was to investigate E-cadherin and alpha-catenin expression in superficial noninvasive papillary TCC and invasive TCC, and correlate these results with pathological and clinical parameters. We have used immunohistochemistry to localize Ecadherin and alpha-catenin in 56 formalin-fixed, paraffin-embedded tissue blocks from 41 patients with superficial bladder cancer and 15 with invasive bladder cancer. The 46 male and 10 female patients had a mean age of 67 years, with range of 40 to 82 years. The mean follow-up time was 33.4 (range 5-120) months. Tumor grade 1:2:3 ratios were 5:32:19. In superficial bladder tumor, abnormal expression of E-cadherin and alpha-catenin was demonstrated in 37 and 71% of the tumors, respectively. In advanced bladder tumor, abnormal expression of E-cadherin and alpha-catenin was demonstrated in 80 and 100% of the tumors, respectively. Differences in expression of E-cadherin and alpha-catenin could be discerned between superficial and advanced bladder tumors (p=0.004, p=0.024, respectively). However, the association between E-cadherin and alpha-catenin expression and tumor grade was not statistically significant (p>0.05). In addition, the expression of E-cadherin and alpha-catenin did not correlate with tumor number and size (p>0.05). We have demonstrated that abnormal expression of E-cadherin and/or alpha-catenin occurs in more than 85% of bladder carcinomas and correlates significantly only with advanced stage. Nevertheless, these observations need to be confirmed in larger prospective clinical studies.

Characterization of Notch Signalling Pathway Members in Normal Prostate, Prostatic Intraepithelial Neoplasia (PIN) and Prostatic Adenocarcinoma.

Prostate Cancer (PCa) holds the second place in terms of cancer-related mortality rate among men. The Notch signalling pathway regulates the proliferation and differentiation in embryonic and adult tissues and determines the cell fate. The body of knowledge in the present literature is currently controversial about the effect of the Notch pathway on prostatic cancer. Therefore, the present study aimed to examine the immunolocalization and expression levels of Notch1-4, Jagged1-2, Delta, HES1 and HES5 from among the members of the Notch signalling pathway in tissues of normal, prostatic intraepithelial neoplasia (PIN) and malignant prostate. The current study included a sample of 20 patients with localised prostatic adenocarcinoma, 18 patients with high grade PIN (H-PIN) and 18 normal prostatic tissue. Immunolocalisations of Notch1, 2, 3, 4, Jagged1, 2, Delta, HES1 and HES5 were identified through the immunohistochemical method. The findings of the present study showed that all in-scope members of the Notch signalling pathway were localised in PIN structures to a greater extent than in other tissues and from amongst these members, specifically Notch1, Notch4, Jagged1 and HES1 were at more significant levels. Consequently, the findings of the present study may indicate that the Notch signalling pathway can play a role especially in the formation of PIN structures.

The assessment of PTEN tumor suppressor gene in combination with Gleason scoring and serum PSA to evaluate progression of prostate carcinoma.

OBJECTIVE: The purpose of the study was to determine if the tumor suppressor gene phosphate and tensin homolog (PTEN) (mutated in multiple advanced cancers 1) in combination with Gleason scoring and serum prostate specific antigen (PSA) could be employed to better predict the progression of prostate carcinoma. MATERIALS AND METHODS: The study group consisted of 43 patients with benign prostate hyperplasia (BPH), 15 with organ confined prostate carcinoma (OCPCa), and 18 with advanced prostate carcinoma (APCa). Prostate tissue samples were obtained from radical prostatectomy, transurethral resection, and TRUS guided trans-rectal needle biopsy and then evaluated for biomarker expression. The clinical stage was assessed according to tumor node metastasis classification and grade according to Gleason system. Serum PSA was measured by conventional techniques and Western blotting analysis was used to determine PTEN expression in the primary tissue. Multivariate analysis was performed to analyze whether these markers could individually predict the progression of prostate carcinoma. RESULTS: APCa patients displayed higher Gleason scores and serum PSA levels. But much lower PTEN expression was detected in prostate of APCa patients compared to patients with BPH or OCPCa. Hormone refractory (HR) and hormone sensitive (HS) APCa cases did not yield any significant differences in terms of Gleason scoring, serum PSA and PTEN expression. PSA levels were significantly higher in patients with OCPCa or APCa compared to patients with BPH. CONCLUSION: Our results suggested that both PTEN and serum PSA appeared to be useful as independent markers to depict the nature of tumor behavior as benign or malign. In addition, PTEN also appeared to be useful as an independent marker to predict the progression of prostate carcinoma.

The potential role of inducible nitric oxide synthase (iNOS) activity in the testicular dysfunction associated with varicocele: an experimental study.

Nitric oxide (NO) has been reported to be increased in the spermatic veins of men affected by varicocele. The aim of the present study was to determine whether iNOS (inducible nitric oxide synthase) has a role in testicular dysfunction associated with varicocele, immunohistochemistry analyze was used to study iNOS activity in testis of adolescent rats with experimental left varicoceles. Rats were randomly divided into three groups. The first group consisted of rats undergoing partial ligation of left renal vein (n:12). The second group consisted of rats undergoing a sham operation (n:6) and, the third group referred to as control rats (n:7). Immunohistochemistry slides were evaluated by counting the number of positive cells and expressed as percents (% iNOS activity). We found that iNOS was predominantly expressed in the cytoplasm of Leydig cells in each group and only a small amount of iNOS was expressed in Sertoli cells. There were significant differences in % iNOS activity between both testes of varicocele group and both of testes control group (p < 0.01), but no significant differences were noted between other groups (p > 0.05). Because of iNOS activity was markedly increased in the Leydig cells of varicocele bearing rats, we suggest that iNOS activity may play a role in the testicular dysfunction associated with varicocele during adolescence.

Sex cord tumour of the adult testis.

Sex cord stromal tumors are rare and account for approximately 6% of all testicular neoplasms. We report a case of sex cord tumor composed of granulosa cells and Sertoli cells in the adult testis.

Clinical pregnancy after uterus transplantation.

OBJECTIVE: To present the first clinical pregnancy after uterus transplantation. DESIGN: Case study. SETTING: Tertiary center. PATIENT(S): A 23-year-old Mayer-Rokitansky-Kuster-Hauser syndrome patient with previous vaginal reconstruction and uterus transplantation. INTERVENTION(S): Eighteen months after the transplant, the endometrium was prepared for transfer of the thawed embryos. MAIN OUTCOME MEASURE(S): Implantation of embryo in an allografted human uterus. RESULT(S): The first ET cycle with one day 3 thawed embryo resulted in a biochemical pregnancy. The second ET cycle resulted in a clinical pregnancy confirmed with transvaginal ultrasound visualization of an intrauterine gestational sac with decidualization. CONCLUSION(S): We have presented the first clinical pregnancy in a patient with absolute uterine infertility after uterus allotransplantation. Although the real success is the delivery of a healthy near-term baby, this clinical pregnancy is a great step forward and a proof of concept that the implantation phase works.

Laparoscopy can aggravate the severity of pancreatitis in an experimental rat model.

PURPOSE: Over the past decade, laparoscopic techniques have markedly evolved, and it has been shown that minimally invasive surgery can provide a safe, effective, and less traumatic management of various surgical diseases. Additionally, it is well known that pancreatitis itself also produced severe oxidative tissue injury by increasing levels of reactive oxygen species. This study therefore aimed to investigate the effects of pneumoperitoneum on the severity of pancreatitis in a rat model of acute pancreatitis induced by cerulein. MATERIALS AND METHODS: Thirty-five Wistar albino rats were divided into five groups with seven rats in each. Experimental pancreatitis was induced using intraperitoneal injection of cerulein. The first group received open laparotomy. Groups 2-5 were treated with 5, 10, 15, and 20 mm Hg, respectively, achieved by applying pressure and waiting for 60 minutes. After this waiting interval, all of the rats were sacrificed; blood samples were taken by intracardiac puncture for biochemical assays, and pancreatic tissue samples were taken for light microscope analysis. Histopathology was scored according to edema, granulation, polymorphonuclear leukocytes, and mononuclear cells in all groups. RESULTS: Great increases in malondialdehyde and reduced glutathione levels were seen in all of the groups in which pancreatitis was induced. In Group 2-5, more significant increases were detected than in the open laparotomy group (P<.005). In the histopathological examination, Groups 2-5 showed more inflammatory cell infiltration, edema, and granulation tissue than the open laparotomy group (P<.005). CONCLUSIONS: It is useful to remember the parameters of the medical treatment of pancreatitis. While surgical treatment is being decided, we think that the process of all kinds of surgery, including laparoscopic surgery, can increase the severity of pancreatitis.

Differential expression of TRAIL and its receptors in benign and malignant prostate tissues.

PURPOSE: Because TRAIL (tumor necrosis factor related apoptosis inducing ligand) selectively kills cancer cells without damaging normal cells, a gene therapy approach using TRAIL is feasible for treating patients with cancer. However, recent publications suggest that significant portions of human tumors appear to be TRAIL resistant. Furthermore, there is some controversy about whether TRAIL receptor composition influences TRAIL sensitivity in cancer cells. Our recent studies suggest that TRAIL receptor composition is the major modulator of TRAIL sensitivity, as demonstrated using prostate, breast and lung cancer cells. We investigated TRAIL and TRAIL receptor expression profiles during prostate carcinogenesis to evaluate their potential as biomarkers and predict the feasibility of a related gene therapy approach. MATERIALS AND METHODS: Paraffin embedded prostate tissues of 44 patients with benign prostatic hyperplasia, 28 with organ confined prostate carcinoma and 26 with advanced prostate carcinoma were analyzed using immunohistochemical staining procedures. RESULTS: Significant levels of TRAIL-R4 decoy receptor expression were detected in patients with benign prostatic hyperplasia, and organ confined and advanced prostate carcinoma. All TRAIL markers tested appear to be valuable markers for separating patients with benign prostatic hyperplasia from patients with organ confined prostate carcinoma or advanced prostate carcinoma. CONCLUSIONS: Due to high TRAIL-R4 expression in all patient groups complementary gene therapy modalities might be needed to bypass potential TRAIL-R4 induced resistance.

A random-pattern skin-flap model in streptozotocin diabetic rats.

BACKGROUND: Flap operations are frequently performed in diabetic patients. Nevertheless, we could find no experimental study examining diabetes mellitus's effect on the flaps' survival. For this reason, we designed this study as a random-pattern skin-flap model of diabetic rats in 1999. METHODS: We used 72 rats weighing about 200 g each. The animals were divided into 2 groups, 1 experimental (diabetic) and 1 control (nondiabetic). Following their diabetic periods, we elevated the rats' modified McFarlane flaps and measured their viable flap areas. RESULTS: The mean percentage of the flap area surviving was 51.40% in the 2-week experimental group. It was 48.20% in the 4-week experimental group and 36.70% in the 8-week experimental group. The mean percentage of flap area surviving was 65.87% in the united control group (the total of all control groups). The mean surviving skin-flap area in the united control group was significantly higher than in the 4- and 8-week experimental groups. Moreover, the mean surviving flap area in the 8-week experimental group was significantly lower than in the 2-week and 4-week experimental groups. CONCLUSIONS: Our study demonstrated that a 4-week diabetic duration for rats is sufficient to observe diabetes' deleterious effects on the flaps' viability. These effects were significantly established, however, after 8 weeks of diabetes. To obtain definitive results, at least 8 weeks of diabetic duration are preferred for similar studies.

Comparison of hypospadiac and normal preputial vascular anatomy.

PURPOSE: Data about the differences between the vascularization of normal and hypospadiac prepuce are lacking. We investigated the course of the preputial arterial blood vessels in normal controls and children with hypospadias by using transillumination, arterial methylene blue injection and 3-dimensional (3-D) reconstruction of serial histological sections focusing on arterial vessels. MATERIALS AND METHODS: Prepuce of 48 normal controls and 15 children with hypospadias was transilluminated by a front and back lighting technique and then photographed. All of the normal and 12 of hypospadiac prepuces not used for urethroplasty or penile body skin reconstruction were removed. The blood vessels of normal prepuce were also identified after arterial injection of methylene blue. Selected prepuce of normal controls and children with hypospadias was serially sectioned, and arterial and venous vessels were histologically distinguished. A 3-D computer reconstruction of the arterial system of normal and hypospadiac prepuces was performed. RESULTS: We confirmed the reliability of the transillumination technique to describe the arterial vascular anatomy of the prepuce by comparing the transillumination to methylene blue injection and 3-D reconstruction of histological sections. We classified the arterial vascular anatomy of normal prepuce as 1 artery predominant (41.67%), 2 arteries predominant (25%), H-type arching artery (12.5%) and net-like arterial system (20.83%). However, hypospadiac prepuce revealed a net-like arterial system more frequently (50%). We noted that the frequency of net-like arterial system was higher in more severe hypospadiac prepuce. CONCLUSIONS: Understanding the differences between normal and hypospadiac prepuce vascular anatomy is germane to hypospadias surgery. The arterial blood supply of the hypospadiac prepuce is different than normal. A better knowledge of the vascular anatomy of the hypospadiac prepuce may improve the surgical results of hypospadias repair.

Antioxidant effect of T-type calcium channel blockers in gastric injury.

It is known that calcium ion has an important role in the cellular function. For this reason, calcium channel blockers may have a protective action against gastric injury which is induced by various stimuli. In this study, the influence of mibefradil on ethanol-induced gastric injury was investigated in rats. Mibefradil was given at a dose 50 mg/kg intraperitoneally 30 min before administration of 1 ml absolute ethanol given by gavage. We compared this effect of mibefradil with that of omeprazol. Ethanol-induced mucosal damage was evaluated using three different approaches: analysis of biochemical parameters and pathologic and macroscopic investigation. It was found that pretreatment with mibefradil significantly reduced ethanol-induced macroscopic, pathologic, and biochemical changes in the gastric mucosa. In conclusion, it is speculated that this findings may prove important in the development of new and improved therapies for the treatment and prevention of gastric ulcers in humans.

Melatonin prevents ethanol-induced gastric mucosal damage possibly due to its antioxidant effect.

Oxygen radical release has been proposed as a pathogenic factor of the ethanol-induced acute gastric injury. Melatonin, a pineal hormone, is known to scavenge oxygen free radicals. We investigated whether parenteral administration of melatonin prevented ethanol-induced macroscopic damage, polymorphonuclear (PMN) leukocyte infiltration, depletion of total glutathione (tGSH) concentration, and glutathione reductase (GSSG-Rd) activity in the rat gastric mucosa. We compared the effects of melatonin with those of omeprazole. Ethanol-induced mucosal damage was evaluated using three different parameters: gastric total glutathione (tGSH) concentration and glutathione reductase (GSSG-Rd) activity, the number of PMN leukocytes, and macroscopic investigation. Gatric tGSH concentration and GSSG-Rd activity decreased and the number of PMNs increased after ethanol administration. It was found that pretreatment with melatonin increased both tGSH concentration and GSSG-Rd activity. Melatonin also reduced ethanol-induced PMN infiltration in the stomach. Ethanol administration damaged the entire gastric mucosa. Melatonin significantly decreased the extent of ethanol-induced macroscopic injury. In conclusion, these findings support the conclusion that the protection conferred by melatonin in gastric ulcer is presumably due to its antioxidant activity.