Effects of chronic stress on bladder morphology of rats and impact of comfort food diet as an ameliorating agent.

OBJECTIVES: To investigate the effects of chronic stress on bladder morphology and the impact of food preference (standard or comfort foods) on the bladder of stressed rats. METHODS: In total, 32 Wistar male rats (3 months old) were divided into four groups: control (C), stressed (S), control + comfort food (C + CF), and stressed + comfort food (S + CF). Groups C and C + CF were maintained under normal conditions, while groups S and S + CF were subjected to chronic stress by the restraint method. Groups C and S received standard rat chow, while groups C + CF and S + CF received comfort food (Froot Loops®) and standard chow. The stress stimuli were induced daily for 2 h over 8 weeks. After 8 weeks, all animals were killed, and the bladders were removed and used for histomorphometric analysis. RESULTS: Body mass was similar among the groups. Stress did not promote differences regarding food intake, but animals receiving comfort food showed higher calories intake (in kcal/Kg) than animals receiving only standard chow. The C + CF and S + CF groups preferred comfort food over the standard chow; this preference was higher in the S + CF than in the C + CF group. The surface density of smooth muscle was reduced in stressed animals, while connective tissue and elastic system fiber content were increased in stressed groups. Further, epithelial height was increased in rats submitted to chronic stress. The surface density of elastic system fibers was decreased by the consumption of comfort food. CONCLUSIONS: Chronic stress induces morphological modifications on the bladder wall and epithelium. These modifications may be related to lower urinary tract symptoms. Additionally, chronic stress caused a higher preference for comfort food intake which did not ameliorate or aggravate the stress-induced bladder alterations.

Quercetin as a nephroprotector after warm ischemia: histomorphometric evaluation in a rodent model

ABSTRACT Purpose: To quantitatively evaluate the possible long-term protective effects of quercetin during renal warm ischemia. Materials and Methods: Male rats were allocated into 4 groups: sham (S), sham quercetin (SQ), ischemia (I), and ischemia quercetin (IQ). Groups SQ and IQ received quercetin (50mg/kg) before and after surgery. Groups I and IQ had their left renal vessels clamped for 60 minutes. All animals were euthanized four weeks after the procedure, and serum urea and creatinine levels were measured. Renal weight and volume, cortex-non-cortex area ratio (C-NC), cortical volume (CV), glomerular volumetric density (Vv[glom]), volume-weighted glomerular volume (VWGV) and number of glomeruli per kidney (N[glom]) were evaluated by stereological methods. Results were considered statistically significant when p <0.05. Results: Serum urea levels in group I increased by 10.4% in relation to group S, but no differences were observed among the other groups. The C-NC of group I was lower than those of all other groups, and group IQ had similar results to sham groups. The Vv[glom] and N[glom] of group I were lower than those of group S (33.7% and 28.3%, respectively) and group IQ had no significant difference compared to the S group. Conclusions: Quercetin was effective as a nephroprotective agent in preventing the glomerular loss observed when the kidney was subjected to warm ischemia. This suggests that this flavonoid may be used preventively in kidney surgery, when warm ischemia is necessary, such as partial nephrectomy.

Quercetin as a nephroprotector after warm ischemia: histomorphometric evaluation in a rodent model.

PURPOSE: To quantitatively evaluate the possible long-term protective effects of quercetin during renal warm ischemia. MATERIALS AND METHODS: Male rats were allocated into 4 groups: sham (S), sham quercetin (SQ), ischemia (I), and ischemia quercetin (IQ). Groups SQ and IQ received quercetin (50mg/kg) before and after surgery. Groups I and IQ had their left renal vessels clamped for 60 minutes. All animals were euthanized four weeks after the procedure, and serum urea and creatinine levels were measured. Renal weight and volume, cortex-non-cortex area ratio (C-NC), cortical volume (CV), glomerular volumetric density (Vv[glom]), volume-weighted glomerular volume (VWGV) and number of glomeruli per kidney (N[glom]) were evaluated by stereological methods. Results were considered statistically significant when p < 0.05. RESULTS: Serum urea levels in group I increased by 10.4% in relation to group S, but no differences were observed among the other groups. The C-NC of group I was lower than those of all other groups, and group IQ had similar results to sham groups. The Vv[glom] and N[glom] of group I were lower than those of group S (33.7% and 28.3%, respectively) and group IQ had no significant difference compared to the S group. CONCLUSIONS: Quercetin was effective as a nephroprotective agent in preventing the glomerular loss observed when the kidney was subjected to warm ischemia. This suggests that this flavonoid may be used preventively in kidney surgery, when warm ischemia is necessary, such as partial nephrectomy.

The bovine kidney as an experimental model: Simulated partial nephrectomy of the cranial pole and proportional analysis of the arterial segments.

Previous studies have shown that the pig kidney is not a good model for some procedures. This study aimed to describe the relationship between the collecting system and the intrarenal arteries, the arterial segments, and to evaluate the bovine kidney as an experimental model for partial nephrectomy of the cranial pole. Polyester resin endocasts of the kidney collecting system together with the intrarenal arteries were prepared. Thirty-two kidneys were used to evaluate the relationship between the collecting system and the intrarenal arteries, while 25 kidneys were transversally sectioned at different points to simulate partial nephrectomy of the cranial pole. Polyester resin of different colors was injected into each segmental artery of the 24 kidneys to evaluate the arterial segments proportionally. The renal artery was divided into cranial and caudal primary branches in 75% of the cases. The cranioventral branch curved on the cranial pole and ran toward the ventral mid-zone in 56.3% of the cases, resembling the retropelvic artery of the human kidney. The kidney was divided into two (25%) or three (75%) arterial segments. The caudal arterial segment had the highest proportional volume (62%). The cranioventral branch was damaged in 28.6% of the kidneys sectioned 1 cm inside the hilum. The arterial branching pattern, the arterial segmentation, and the impairment of the arterial supply after the simulated partial nephrectomy of the cranial pole are quite different from those found in humans. Thus, all differences should be taken into account when using the bovine kidney as a model.

Estudos anteriores demonstraram que o rim do porco não é um bom modelo para alguns procedimentos. O objetivo deste estudo foi descrever a relação entre o sistema coletor e as artérias intrarrenais, os segmentos arteriais, e avaliar o rim bovino como modelo experimental para a nefrectomia parcial do polo cranial. Foram confeccionados moldes de resina de poliéster do sistema coletor renal associado às artérias intrarrenais. Para a avaliação da relação entre o sistema coletor e as artérias intrarrenais foram utilizados trinta e dois rins, enquanto outros trinta e cinco rins foram cortados transversalmente em diferentes regiões para simular a nefrectomia parcial do polo cranial. Resina de poliéster de diferentes cores foi injetada em cada segmento arterial de 24 rins para avaliar proporcionalmente os segmentos arteriais. A artéria renal se dividiu em ramos primários cranial e caudal em 75% dos casos. O ramo cranioventral contornou o polo cranial e seguiu para a região média ventral em 56,3% dos casos, lembrando a artéria retropiélica do rim humano. O rim estava dividido em dois (25%) ou três (75%) segmentos arteriais. O segmento arterial caudal apresentou o maior volume proporcional (62%). O ramo cranioventral foi lesionado em 28,6% dos rins seccionados 1 cm internamente ao hilo. O padrão da ramificação arterial, os segmentos arteriais, e o dano ao suprimento arterial após a nefrectomia simulada parcial do polo cranial são bem diferentes daqueles encontrados em humanos. Então, todas as diferenças devem ser levadas em consideração quando se usa o rim bovino como modelo.

Anatomical background of ovine kidney for use as animal model: analysis of arterial segmentation, proportional volume of each segment and arterial injury after cranial pole partial nephrectomy

ABSTRACT Objective To study the arterial segments of ovine kidney, present a proportional volume analysis of each kidney arterial segment, and analyze arterial injuries caused by simulated partial nephrectomy of cranial pole. Materials and Methods Forty-eight ovine kidneys injected with polyester resin into the renal arteries and collecting system were used in this study. Eighteen kidneys were used to study the arterial segments and the proportional volume of each renal segment. Other 30 kidneys were submitted to superior pole resection at a distance of 1.0cm, 0.5cm, or exactly at the cranial hilar edge, just before the resin hardening. These endocasts were used to evaluate the arterial injuries caused by these different resection planes. Results Ovine renal artery divided into two (ventral and dorsal) or three segmental arteries. Dorsal segment presented higher proportional volume than ventral segment. For kidneys with three segments, the third segment was on the caudal region (caudo-ventral or caudo-dorsal segment) and presented the lowest proportional volume. None of the resected kidneys (at 1.0, 0.5 or at the cranial hilar edge) presented injury of arterial branches that irrigate non-resected region. Conclusion The segmental distribution of renal artery, the proportional volume of each segment and arterial injuries after cranial pole resection in ovine kidneys are different from what is observed in human kidneys. Meanwhile, ovine kidneys show a primary segmental division on anterior and posterior, as in humans, but different from swine. These anatomical characteristics should be considered when using ovine as animal models for renal experimental and/or training procedures.

Anatomical background of ovine kidney for use as animal model: analysis of arterial segmentation, proportional volume of each segment and arterial injury after cranial pole partial nephrectomy.

OBJECTIVE: To study the arterial segments of ovine kidney, present a proportional volume analysis of each kidney arterial segment, and analyze arterial injuries caused by simulated partial nephrectomy of cranial pole. MATERIALS AND METHODS: Forty-eight ovine kidneys injected with polyester resin into the renal arteries and collecting system were used in this study. Eighteen kidneys were used to study the arterial segments and the proportional volume of each renal segment. Other 30 kidneys were submitted to superior pole resection at a distance of 1.0cm, 0.5cm, or exactly at the cranial hilar edge, just before the resin hardening. These endocasts were used to evaluate the arterial injuries caused by these different resection planes. RESULTS: Ovine renal artery divided into two (ventral and dorsal) or three segmental arteries. Dorsal segment presented higher proportional volume than ventral segment. For kidneys with three segments, the third segment was on the caudal region (caudo-ventral or caudo-dorsal segment) and presented the lowest proportional volume. None of the resected kidneys (at 1.0, 0.5 or at the cranial hilar edge) presented injury of arterial branches that irrigate non-resected region. CONCLUSION: The segmental distribution of renal artery, the proportional volume of each segment and arterial injuries after cranial pole resection in ovine kidneys are different from what is observed in human kidneys. Meanwhile, ovine kidneys show a primary segmental division on anterior and posterior, as in humans, but different from swine. These anatomical characteristics should be considered when using ovine as animal models for renal experimental and/or training procedures.

Histomorphometric and biochemical data of rat kidney submitted to warm ischemia associated with resveratrol treatment.

The data presented here come from the article "Histomorphometric evaluation of the rat kidney submitted to warm ischemia and the protective effect of resveratrol" [1]. Rats of Wistar lineage (n = 39; 9 weeks of age) were obtained and apportioned into 4 groups at random. Both groups Sham (S) and Sham Resveratrol (SR) were submitted to open laparotomy and dissection of the left renal pedicle, the same as groups Ischemia (I) and Ischemia Resveratrol (IR), being the last two also submitted to 1 h left warm renal ischemia. SR and IR were treated with 30 mg/kg of resveratrol intraperitoneally 1 h before the surgical procedure, while S and I received saline injections. Rats were killed a month after surgery by anesthetic overdose. A blood sample was collected by cardiac puncture for determination of serum urea and creatinine serum by biochemical analysis at automated enzymatic method. Kidneys were weighted, Sherle´s method was used for measurement of their volume and then both were fixated in buffered formalin for 48 h. Cortex-non-cortex areas ratio (C-NC) was assessed by Cavalieri's method using a stereoscope. The product of multiplying the renal volume by the C-NC is the cortical volume (CV). Left kidneys fragments were processed for histology resulting in slides that were stained with haematoxylin and eosin. For histomorphometric analyses, 25 random cortical fields were photographed at 200x magnification using a camera attached to a light microscope. The estimation of glomerular volumetric density (Vv [Glom]), indication of proportional volume occupied by glomeruli in the cortex, was performed by the point-counting method. The point-sampled intercepts method was used to estimate the volume-weighted mean glomerular volume (VWGV). Total number of glomeruli per kidney (N [Glom]) estimation was achieved through the formula CVxVv [Glom]/VWGV. All the data were tabulated in spreadsheets. The quantitative results were compared by one-way ANOVA with Tukey's post-test using GraphPad Prism software. All results were considered significant when the value of p <0.05.

Histomorphometric evaluation of the rat kidney submitted to warm ischemia and the protective effect of resveratrol.

BACKGROUND: The objective of this study is to quantitatively evaluate the protective effects of resveratrol for using during renal warm ischemia. METHODS: Rats were allocated into 4 groups: Sham, Sham Resveratrol, Ischemia, Ischemia Resveratrol. Sham Resveratrol and Ischemia Resveratrol received resveratrol before surgery. Ischemia and Ischemia Resveratrol had renal vessels clamped. Animals were euthanized four weeks after. Serum urea and creatinine were measured. Renal weight and volume, cortex-non-cortex areas ratio, cortical volume, glomerular volumetric density, volume-weighted mean glomerular volume and number of glomeruli per kidney were evaluated. RESULTS: Serum urea in Ischemia increased by 10.4% compared to Sham and no differences were observed among Ischemia Resveratrol and sham groups. The glomerular volumetric density and number of glomeruli of Ischemia were lower than Sham but Ischemia Resveratrol had no difference compared to sham groups. CONCLUSIONS: Preoperative administration of resveratrol has renoprotective effects, preventing the glomerular number reduction observed in warm ischemia.

Antioxidants as Renoprotective Agents for Ischemia during Partial Nephrectomy.

Small renal masses have been diagnosed increasingly in recent decades, allowing surgical treatment by partial nephrectomy. This treatment option is associated with better renal function preservation, in comparison with radical nephrectomy. However, for obtaining a bloodless field during surgery, occlusion of renal artery and veins is often required, which results in transitory ischemia. The renal ischemia-reperfusion injury is associated with increased reactive oxygen species production leading to renal tissue damage. Thus, the use of antioxidants has been advocated in the partial nephrectomy perioperative period. Several antioxidants were investigated in regard to renal ischemia-reperfusion injury. The present manuscript aims to present the literature on the most commonly studied antioxidants used during partial nephrectomy. The results of experimental and clinical studies using antioxidants during partial nephrectomy are reported. Further, alimentary sources of some antioxidants are presented, stimulating future studies focusing on perioperative antioxidant-rich diets.

Tannic Acid Solution: A Better Fixative Solution Than Formalin for Elastin and Collagen-Toxic and Morphological Assessment.

Formaldehyde is commonly used worldwide, even though it is classified as carcinogenic to humans by the International Agency for Research on Cancer. This has motivated intensive investigations of formaldehyde substitutes, and recently, some alternative solutions were found, which can potentially replace it. Previous research showed that tannic acid (TA) in glutaraldehyde solution has the ability to stabilize elastin and collagen. This provided a basis for the development of a new alcoholic fixative solution, particularly aimed at extracellular matrix components, with TA as a main component. Heart, brain, and intestinal samples were fixed by immersion in 10% regular formalin solution (RFS), 70% ethanol solution (ES), and tannic acid ethanolic solution (TAES). Next, tissue fragments were prepared for routine histology procedures. The toxicity of TA was analyzed using in silico tests for mutagenicity, as well as for cutaneous and respiratory toxicity. Analyses of photomicrographs demonstrated that all fixative solutions have the ability to preserve the fragments. The quantitative analyses showed that capability of TAES to preserve and stabilize elastin and collagen is superior to that of RFS and ES. We demonstrated that TA is not mutagenic, and it is less toxic for skin and respiratory tract. We therefore conclude that TAES can potentially represent a powerful and feasible alternative solution for fixing extracellular matrix of microscopic examination samples. Anat Rec, 301:1544-1550, 2018. © 2018 Wiley Periodicals, Inc.

Mannitol reduces nephron loss after warm renal ischemia in a porcine model.

BACKGROUND: Mannitol has been employed to ameliorate renal warm ischemia damage during partial nephrectomy, however, there is limited scientific evidence to support the use of mannitol during partial nephrectomy. The objective of the present study was to investigate the glomerular number after renal warm ischemia, with and without the use of mannitol in a Pig Model. METHODS: Twenty-four male pigs were assigned into three groups. Eight animals were allocated to the sham group that was subjected to laparoscopic dissection of the left renal hilum, without renal ischemia. Eight animals were allocated to the ischemia group that had the left renal hilum clamped for 30 min through laparoscopic access. Eight animals received mannitol (250 mg/kg) before the occlusion of renal hilum for 30 min. The kidneys were collected after the euthanasia of the pigs 21 days post surgery. The right kidney was utilized as a self-control for each animal. Serum creatinine, urea levels, the weight and volume of the kidneys were measured. Glomerular volumetric density, volume-weighted glomerular volume, and cortical volume were quantified through stereological methods and employed to determine the number of nephrons per kidney. Student's t test and ANOVA were used for statistical analysis. RESULTS: In the ischemia group, the left kidney recorded a reduction of 24.6% (290, 000 glomeruli) in the number of glomeruli in comparison to the right kidney. Kidneys subjected to ischemia also displayed decreased weight and volume in comparison to the sham and mannitol groups. No difference was observed between the left and right kidneys from the sham and mannitol groups. Further, no distinction in serum creatinine and urea among the groups was observed. CONCLUSION: The use of mannitol significantly reduces nephron loss during warm ischemia in pigs.

Does a Ureteral Obstruction Affect the Contralateral Kidney Morphology? A Stereological Analysis in a Rodent Model.

OBJECTIVE: To investigate, in a rodent model, the influence of an obstructed kidney/ureter on the contralateral organ morphology. METHODS: Thirty-six rats were randomly assigned into the following groups: untreated ureteral obstruction group (UO-U, n = 10), submitted to a ligature of the ureter at day 0; UO group (at day 0) followed by nephrectomy after 24 h (UO-N, n = 8); nephrectomy group (N, n = 9), submitted to nephrectomy at day 0; and sham group (S, n = 9), submitted to simulated surgery at day 0. All these procedures were performed on the left kidney/ureter. All animals were euthanized 30 days after surgery, and the right kidneys were collected for stereological analysis. Data were compared using a one-way analysis of variance with Bonferroni's post test, considering p < 0.05 as significant. RESULTS: The kidney volume, weight, and cortical volume were augmented in groups UO-N and N, compared to group S. Kidneys from the UO-U group showed only a moderate augmentation in kidney weight, with a reduction in the cortical-medullar ratio, compared to the kidneys in group S. No differences in the glomerular volumetric density, volume-weighted glomerular volume, and number of glomeruli were observed among the right kidneys of the studied groups. CONCLUSION: An obstructed kidney/ureter is more prejudicial to the contralateral kidney morphology than an absent kidney.

Immediate and late effects of chronic stress in the testes of prepubertal and adult rats.

The objective of this study was to investigate the effects of chronic stress on the testes of prepubertal and adult rats and to evaluate whether any alterations could be reversed when stress induction is ended. Seventy-six male rats were assigned to eight groups depending on the type of treatment (control or stressed), the age at which stress was initiated (prepubertal or adult), and the time of evaluation (immediate or late). Stress stimuli were applied for 6 weeks. Stressed prepubertal and adult rats evaluated immediately after the last stress stimulus were included in SP-I and SA-I groups, respectively. The late prepubertal (SP-L) and adult (SA-L) groups of stressed rats were evaluated 6 weeks after the last stress stimulus. Age-matched rats were used as controls (CP-I, CA-I, CP-L, and CA-L groups). Application of stress stimuli to rats in the SP-I group resulted in body weight and seminiferous tubule diameter reduction. The rats in the SA-I group also showed several functional (testosterone level and sperm parameter) and morphological (testicular weight and seminiferous tubule diameter) reductions. The rats in the SP-L group showed increased body weight and intertubular compartment volumetric and absolute densities and reduced tubular compartment volumetric density. The rats in the SA-L group presented only reduced sperm viability. Stress stimuli promoted changes in the rats in all the study groups. The testes of the adult rats were the most affected by chronic stress. However, the stressed adult rats recovered well from the testicular alterations.

Glomerular Morphology and Renal Function after Warm Ischemia by Main Artery or Selective Clamping in a Porcine Model.

OBJECTIVE: The study aimed to evaluate the ischemic and non-ischemic areas after selective arterial occlusion by using stereological analysis of glomeruli, and to compare them with main arterial clamping and sham-operated animals. MATERIALS AND METHODS: Twenty-four male pigs were used in the study. The animals were divided into 3 groups with 8 animals in each as follows: group sham, submitted to laparoscopic dissection of the renal pedicle but not submitted to ischemia; group arterial (A), submitted to left renal artery clamping; and group selective (S), submitted to left renal artery caudal branch occlusion. Groups A and S underwent 30 min of warm ischemia. Left and right kidneys were collected after 21 days and renal fragments were processed for stereological evaluation. Glomerular volume density (Vv[glom]), mean glomerular volume (MGV), and glomerular density were measured. Serum creatinine and urea were assessed preoperatively, 10 days after surgery, and before euthanasia. RESULTS: There was no significant difference among groups with regard to renal function. Renal weight and volume were similar among groups. Also, no difference was observed between the groups with regard to Vv[glom], MGV, and glomerular density, both when compared to its right control or when left kidneys were compared. CONCLUSIONS: Selective arterial clamping technique was neither superior nor inferior to main artery clamping.

Effects of castration on penile extracellular matrix morphology in domestic cats.

Objectives This study was undertaken to verify the possible modifications caused by hormonal deprivation in the extracellular matrix in the penises of neutered cats. Methods Twenty-seven penises from domestic shorthair cats were collected: 14 samples from intact cats and 13 from neutered cats. Sections were stained with Weigert's resorcin-fuchsin, hematoxylin and eosin, and picrosirius red. Histomorphometric analysis was performed using light microscopy and image analysis software. The following parameters were analyzed: density of the elastic fibers and collagen fibers in the corpus spongiosum; density of the elastic fibers in the tunica albuginea of the corpus cavernosum and the tunica albuginea of the corpus spongiosum; luminal area of the urethra; area of the corpus spongiosum; area of the corpus cavernosum; and thickness of the urethral epithelium. The data were analyzed using the Shapiro-Wilk test to verify the normal distribution, and groups were compared using Student's t-test; P <0.05 indicated statistically significant differences. Results Significant differences were observed between intact cats and neutered cats in the density of elastic fibers in the tunica albuginea of the corpus cavernosum (8.13% ± 1.38% vs 3.11% ± 0.66%), tunica albuginea of the corpus spongiosum (4.37% ± 1.08% vs 3.30% ± 1.01%) and corpus spongiosum (6.28% ± 3.03% vs 4.10% ± 2.19%), and density of collagen fibers in the corpus spongiosum (34.11% ± 10.86% vs 44.21% ± 12.72%). Conclusions and relevance The results show a significant decrease in the density of the elastic fibers and a significant increase of the density of the collagen fibers in the corpus spongiosum in neutered animals. This suggests that the compliance of the periurethral region is reduced, and these changes could be a predisposing factor for urethral obstructive disease.

Histomorphometric comparison of the human, swine, and ovine collecting systems.

The ovine kidney has been recently determined to be a better model than the swine kidney for the study of collecting system healing after partial nephrectomy. However, there is no histological study comparing the collecting systems of these species. To compare human, swine, and ovine collecting systems using histomorphometry. The collecting systems of 10 kidneys from each species (human, swine, and ovine) were processed for histomorphometry. The thickness of the three layers (mucosal connective tissue, submucosal muscular, and adventitial connective tissue) were measured. The densities of smooth muscle fibers, elastic system fibers, and cells were also measured. Additionally, blood vessel density in the adventitial connective tissue was measured. Analysis of the collecting systems from the three species presented several differences. The adventitial connective tissue from the swine samples was thicker, with more blood vessels and smooth muscle fibers, compared with that from the human and ovine samples. Swine also had higher density of elastic fibers on the submucosal muscular layer. Ovine and human collecting systems shared several similar features, such as blood vessel and elastic fiber density in all layers and the density of cellular and muscular fibers in the submucosal muscular and adventitial connective tissue layers. The collecting system of the ovine kidney is more similar to that of the human kidney compared with that of the swine kidney. This may explain the differences between the healing mechanisms in swine and those in humans and sheep after partial nephrectomy. Anat Rec, 299:967-972, 2016. © 2016 Wiley Periodicals, Inc.

The sheep as a model for healing studies after partial nephrectomy.

BACKGROUND: The pig has been considered the best model for renal surgery. However, recent research has demonstrated that the kidney of pigs heals differently from that of humans. The objective of this study was to evaluate sheep as an alternative animal model for studying collecting system healing after laparoscopic partial nephrectomy. MATERIALS AND METHODS: The caudal pole of the left kidney was removed from eight female adult domestic sheep using laparoscopic partial nephrectomy. Monopolar energy was used for hemostasis only in the parenchyma, avoiding coagulation near the collecting system, which was left opened. After 14 d, all animals were euthanized, and their left kidney was removed. Serum levels of urea and creatinine were assessed preoperatively and postoperatively (on days 2, 6, 10, and 14), and peritoneal fluid samples were collected during necropsy for urea and creatinine evaluation. An ex vivo retrograde pyelogram was performed, and a retrograde injection of methylene blue ink was administered to evaluate urinary leakage. Samples from the operated pole were analyzed using histologic methods. RESULTS: During necropsy, an urinoma surrounding the operated kidney was observed in one animal. Peritoneal fluid levels of urea and creatinine were elevated. Retrograde pyelograms exhibited contrast-medium extravasation through the operated pole in all kidneys. The opened collecting system was also confirmed by methylene blue ink injection. The operated pole was covered by collagenous tissue and adhered to adjacent organs. CONCLUSIONS: Sheep should be considered as an adequate experimental model for research on collecting system healing after partial nephrectomy.

Effects of testicular transfixation on seminiferous tubule morphology and sperm parameters of prepubertal, pubertal, and adult rats.

Orchiopexy is performed as part of cryptorchidism and testicular torsion treatment. The inflammation caused by the needle and suture penetration has been suggested to be one of the possible causes of subfertility after parenchymal transfixation of the testicles. The purpose of the present study was to investigate testicular alterations after parenchymal transfixation sutures at different ages in rats. Prepubertal, pubertal, and adult rats were submitted to parenchymal suturing (without tying the knots, thus avoiding local ischemic injury) of the right testicle, which was maintained for 4 hours. All animals were subjected to euthanasia on completion of 14 weeks of life. The right testicles were studied as the sutured testicles, whereas the left organs were studied as contralateral. One age-matched control group of rats that was not submitted to any procedure was used for comparison. During euthanasia, sperm were collected from the tail of the epididymal and evaluated for concentration, motility, and viability. Samples from testicular tissue were collected for morphologic analysis. Sperm analysis indicated that only the adult operated animals presented reductions in motility (38.2% of adult vs. 54.1% of control; P = 0.02) and viability (16.6% of adult vs. 24.6% of control; P = 0.003). Several morphologic alterations were noted both in sutured and in contralateral testes at all ages. For instance, the seminiferous epithelium volumetric density of right testicles was reduced from 50.4% in controls to 32.3% in prepubertal operated animals, 45.3% in pubertal operated animals, and 39.4% in adult operated animals (P < 0.05). The seminiferous epithelium volumetric density was also reduced to 39.9% and 39.0% in contralateral testicles of animals operated before and after puberty, respectively (P < 0.05). The animals operated on before puberty and in adulthood showed more testicular morphologic alterations, as seminiferous tubule volumetric density, seminiferous tubule length, and tubular diameter were reduced only in prepubertal and/or adult operated animals. Testicular transfixation in rats led to important morphologic modifications in the ipsilateral and contralateral organs. These alterations were observed regardless of the age when surgery was performed, but they were milder in animals operated on during puberty. Orchiopexy techniques that do not involve the application of testicular transfixation sutures should be recommended.

Effect of antioxidants on outcome of testicular torsion in rats of different ages.

PURPOSE: We assessed reproductive and testicular function in adult rats after testicular torsion created before, during and after puberty, and with vs without resveratrol or arginine treatment. MATERIALS AND METHODS: Age matched rats were divided into groups, including simulated surgery without testicular torsion, 720-degree testicular torsion for 4 hours, testicular torsion with resveratrol treatment and testicular torsion with arginine treatment. To study reproductive function at age 12 weeks each rat mated with 3 females. The males were sacrificed at age 14 weeks. Spermatozoids were collected from the epididymal tail and evaluated for concentration, motility and viability. Testicular samples were collected for morphological analysis. RESULTS: Reproductive function was not altered by testicular torsion but antioxidants improved potency. Compared to sham operated and contralateral samples all spermatozoid parameters from testicular torsion samples were inferior. Resveratrol and arginine did not improve spermatozoid quality or quantity in torsed testes but contralateral samples were improved by each drug. The seminiferous epithelium of rats submitted to testicular torsion during puberty was least affected. Each antioxidant partially to totally prevented the morphological alterations found in rats with untreated testicular torsion. Rats submitted to testicular torsion before puberty that were treated with antioxidants showed the fewest changes. CONCLUSIONS: Testicular morphology was altered less in rats when torsion occurred earlier in life, that is during puberty. Treatment with antioxidants improved contralateral spermatozoid production and some fertility parameters. Each antioxidant also prevented testicular morphology alterations after testicular torsion. Prepubertal rats benefited most from antioxidant treatment.