Impact of maternal stress on metabolism and penile morphology in young offspring rats.

Exposure to prolonged stress in pregnancy and/or lactation can lead to the future development of diseases. We aimed to study the effects of maternal stress on the biometry, metabolism, and penile morphology of young Wistar rats. Animals were divided into two experimental groups: Control Group (C) - pups from control mothers, without any intervention (n=5); and Chronic Stress Group (S) - pups from mothers who suffered variable stress in the third week of pregnancy (14th to 21st day; n=5). Food intake and body mass of the pups (n=10, in the C group and n=9 in the S group) were checked; at euthanasia (three months old), fat deposits and penis were removed. At birth and weaning, S animals were lighter than C animals, [-33.72% (p=0.0422) and -17.07% (p=0.0018)], respectively. However, the final body mass and body mass delta showed no differences. Food intake and fat deposits also did not differ. However, the S group was hyperglycemic at 30 and 60 days of life [+20.59% (p=0.0042) and +14.56% (p=0.0079), respectively], despite the glycemia measured at 90 days showing no difference between groups. Penile areas and surface densities of the corpora cavernosa components were similar between groups. The results indicate that maternal stress is an important metabolic programmer, which generates low birth weight and accelerated recovery of body mass after birth (catch-up). However, in an early analysis (90 days of life), exposure to gestational stress did not change the morphology of the offspring's penis in adulthood.

Glycine disrupts myelin, glutamatergic neurotransmission, and redox homeostasis in a neonatal model for non ketotic hyperglycinemia.

Non ketotic hyperglycinemia (NKH) is an inborn error of glycine metabolism caused by mutations in the genes encoding glycine cleavage system proteins. Classic NKH has a neonatal onset, and patients present with severe neurodegeneration. Although glycine accumulation has been implicated in NKH pathophysiology, the exact mechanisms underlying the neurological damage and white matter alterations remain unclear. We investigated the effects of glycine in the brain of neonatal rats and MO3.13 oligodendroglial cells. Glycine decreased myelin basic protein (MBP) and myelin-associated glycoprotein (MAG) in the corpus callosum and striatum of rats on post-natal day (PND) 15. Glycine also reduced neuroglycan 2 (NG2) and N-methyl-d-aspartate receptor subunit 1 (NR1) in the cerebral cortex and striatum on PND15. Moreover, glycine reduced striatal glutamate aspartate transporter 1 (GLAST) content and neuronal nucleus (NeuN), and increased glial fibrillary acidic protein (GFAP) on PND15. Glycine also increased DCFH oxidation and malondialdehyde levels and decreased GSH concentrations in the cerebral cortex and striatum on PND6, but not on PND15. Glycine further reduced viability but did not alter DCFH oxidation and GSH levels in MO3.13 cells after 48- and 72-h incubation. These data indicate that impairment of myelin structure and glutamatergic system and induction of oxidative stress are involved in the neuropathophysiology of NKH.

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.

What is the effect of Tribulus terrestris on testicular morphology and sperm production? An experimental study in the rat model.

The objective of this study was to investigate whether treatment with Tribulus terrestris (Tt) has any impact on the testicular morphology and function in a rodent model. Twenty male rats were divided into a control group and a group receiving 100 mg kg-1 body weight of Tt supplementation. After 40 days of experiment, the animals were submitted to euthanasia; epididymal tail spermatozoa were collected; and spermatozoa concentration, motility, and viability were analyzed. In addition, testicles were collected and processed for histomorphometrical analyses. Data were compared using the Student's t-test and considered significant when P < 0.05. Spermatozoa concentration, motility, and viability showed no difference between the groups. Further, testicular weight and volume, seminiferous tubule diameter, tunica propria surface density, seminiferous epithelium surface density, and intertubular compartment surface density were statistically similar between the groups. However, seminiferous epithelium height and tubular lumen surface density were augmented in animals treated with Tt. Treatment with Tt does not cause a major impact on testicular morphology, promoting only subtle modifications. No difference on spermatozoa parameters was observed.

Effects of comfort food diet on the penile morphology of stressed rats.

Purpose: To investigate the effects of chronic stress, associated or not with comfort food, on the morphology of the penis. Materials & methods: Thirty-two adult Wistar rats were divided into four experimental groups: Control group (C), receiving standard rat chow, and under normal conditions; Stressed group (S), receiving standard chow, and submitted to stressful situations; Control + comfort food group (C + CF), receiving standard chow and comfort food, and under normal conditions; and Stressed + comfort food group (S + CF), receiving standard chow and comfort food, and submitted to stressful situations. At 10 weeks of age, food supply and stress were initiated. All groups had ad libitum access to standard chow and water, and groups receiving comfort food also had access to Froot Loops®. Chronic stress was induced by restriction, animals were contained daily in polypropylene tubes for 2 h, for eight weeks. After eight weeks all animals were killed; penises were removed for histomorphometric analysis. Results: Body mass was similar among the groups. Food intake in S + CF group was lower than in other groups. Concerning food preference, groups C + CF and S + CF preferred comfort food over the standard chow, with this preference being higher in S + CF than in C + CF. The area of the corpora cavernosa without tunica albuginea was lower in group S + CF than in group C. Most interestingly, the surface density of connective tissue in the corpora cavernosa was higher in groups S and S + CF compared to group C. In contrast, smooth muscle surface density was markedly lower in S + CF compared to groups C and C + CF, while group S also had reduced smooth muscle in comparison to group C. Conclusion: Chronic stress caused a morphological alteration on penile histomorphometry. Also, stress increased the preference for comfort foods which caused more deleterious effects in some parameters.

Effects of dutasteride and tamsulosin on penile morphology in a rodent model.

PURPOSE: To evaluate the penile morphology after the isolated and combined administration of dutasteride and tamsulosin in a rodent model. MATERIALS AND METHODS: Forty male rats were assigned into the following groups: Control group (C, receiving distilled water, n=10); Dutasteride group (D, receiving 0.5 mg/Kg/day of dutasteride, n=10); Tamsulosin group (T, receiving 0.4 mg/Kg/day of tamsulosin, n=10); and Dutasteride associated with Tamsulosin group (DT, receiving both drugs n = 10). All drugs were administered via oral gavage. After 40 days, the animals were submitted to euthanasia and their penises were collected for histomorphometric analyses. Data were compared using one-way ANOVA followed by Bonferroni's post-test, considering p<0.05 as significant. RESULTS: The sinusoidal space and smooth muscle fiber surface densities (Sv), and the cross-sectional penile areas of rats in groups D, T and DT were reduced in comparison to controls with the most notable reductions in the combined therapy group. The connective tissue and elastic system fibers Sv were augmented in groups D, T and DT in comparison with the control group, again with the most pronounced changes observed in animals receiving the combined therapy. CONCLUSION: Both treatments with dutasteride or tamsulosin promoted penile morphometric modifications in a rodent model. The combination therapy resulted in more notable modifications. The results of this study may help to explain the erectile dysfunction observed in some men using these drugs.

Monocrotaline induces acutely cerebrovascular lesions, astrogliosis and neuronal degeneration associated with behavior changes in rats: A model of vascular damage in perspective.

Pyrrolizidine alkaloids (PAs) are secondary plant metabolites playing an important role as phytotoxins in the plant defense mechanisms and can be present as contaminant in the food of humans and animals. The PA monocrotaline (MCT), one of the major plant derived toxin that affect humans and animals, is present in a high concentration in Crotalaria spp. (Leguminosae) seeds and can induce toxicity after consumption, characterized mainly by hepatotoxicity and pneumotoxicity. However, the effects of the ingestion of MCT in the central nervous system (CNS) are still poorly elucidated. Here we investigated the effects of MCT oral acute administration on the behavior and CNS toxicity in rats. Male adult Wistar were treated with MCT (109 mg/Kg, oral gavage) and three days later the Elevated Pluz Maze test demonstrated that MCT induced an anxiolytic-like effect, without changes in novelty habituation and in operational and spatial memory profiles. Histopathology revealed that the brain of MCT-intoxicated animals presented hyperemic vascular structures in the hippocampus, parahippocampal cortex and neocortex, mild perivascular edema in the neocortex, hemorrhagic focal area in the brain stem, hemorrhage and edema in the thalamus. MCT also induced neurotoxicity in the cortex and hippocampus, as revealed by Fluoro Jade-B and Cresyl Violet staining, as well astrocyte reactivity, revealed by immunocytochemistry for glial fibrillary acidic protein. Additionally, it was demonstrated by RT-qPCR that MCT induced up-regulation on mRNA expression of neuroinflammatory mediator, especially IL1ß and CCL2 in the hippocampus and cortex, and down-regulation on mRNA expression of neurotrophins HGDF and BDNF in the cortex. Together, these results demonstrate that the ingestion of MCT induces cerebrovascular lesions and toxicity to neurons that are associated to astroglial cell response and neuroinflammation in the cortex and hippocampus of rats, highlighting CNS damages after acute intoxication, also putting in perspective it uses as a model for cerebrovascular damage.

Stress Hormone Corticosterone Controls Metabolic Mitochondrial Performance and Inflammatory Signaling of In Vitro Cultured Sertoli Cells.

Stress, as a physiological response, is a major factor that affects several processes, including reproductive functions. The main hormonal players of stress are cortisol (humans) and corticosterone (rodents). Sertoli cells (SCs), as key contributors for the testicular homeostasis maintenance, are extensively challenged by different hormones, with glucocorticoid corticosterone being the signaling modulator that may impact these cells at different levels. We aimed to characterize how corticosterone modulates SCs energy balance, putting the mitochondrial performance and signaling output in perspective as the cells can disperse to the surroundings. TM4 mouse SCs were cultured in the absence and presence of corticosterone (in nM: 20, 200, and 2000). Cells were assessed for extracellular metabolic fluxes, mitochondrial performance (cell respirometry, mitochondrial potential, and mitochondrial complex expressions and activities), and the expression of androgen and corticosteroid receptors, as well as interleukine-6 (IL-6) and glutathione content. Corticosterone presented a biphasic impact on the extracellular fluxes of metabolites. Low sub-physiological corticosterone stimulated the glycolytic activity of SCs. Still, no alterations were perceived for lactate and alanine production. However, the lactate/alanine ratio was decreased in a dose-dependent mode, opposite to the mitochondrial complex II activity rise and concurrent with the decrease of IL-6 expression levels. Our results suggest that corticosterone finely tuned the energetic profile of mouse SCs, with sub-physiological concentrations promoting glycolytic expenditure, without translating into cell redox power and mitochondrial respiratory chain performance. Corticosterone deeply impacted the expression of the pro-inflammatory IL-6, which may alter cell-to-cell communication in the testis, in the last instance and impact of the spermatogenic performance.

Amyloid-ß Processing in Aged S100B Transgenic Mice Is Sex Dependent.

(1) Background: Calcium-binding protein S100B is involved in neuroregeneration but has also been associated with neurodegeneration. These contrasting effects may result from concentration or duration of exposure. We investigated the effect of long-term increased S100B levels on amyloid-ß processing in one-year-old transgenic (tg) mice with 12 copies of the murine S100B gene with specific consideration of sex and specific brain regions. (2) Methods: S100B and amyloid-ß 42 (Aß42) were quantified in serum, cerebrospinal fluid (CSF), adipose tissue, and different brain regions by ELISA in wild-type (wt) and S100Btg mice (each n = 7 per group). Thioflavin T (ThT) and Aß immunostaining were performed for visualization of Aß deposition. (3) Results: S100B in serum, CSF, and brain was significantly increased in S100Btg mice of both sexes. Aß42 was significantly increased in the hippocampus of male S100Btg mice (p = 0.0075), and the frontal cortex of female S100Btg mice (p = 0.0262). ThT and Aß immunostaining demonstrated Aß deposition in different brain regions in S100Btg mice of both sexes and female wt. (4) Conclusion: Our data validate this experimental model for studying the role of S100B in neurodegeneration and indicate that Aß processing is sex-dependent and brain region-specific, which deserves further investigation of signaling pathways and behavioral responses.

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.

A toxicological comparison between two uranium compounds in Artemia salina: Artificial seawater containing CaCO3.

Uranium (U) mining is an aquatic environmental concern because most of these harmful compounds are discharged into freshwater, reaching the saline environment as the final destination of this contaminated water. Carbonates are present in ocean waters and are essential for benthic organisms, however they may influence the U-induced toxicity. Thus, the aim of this study was to compare the toxicity of uranium nitrate (UN) and uranium acetate (UA) in Artemia salina (AS), which is one of the leading representatives of the marine biota. The cultures of AS (instar II) maintained in artificial seawater containing CaCO3 were exposed for 24 h to different concentrations of U compounds. The results showed that AS were more sensitive to UN (LC50 ≈ 15 µM) when compared with UA (LC50 ≈ 245 µM) indicating higher toxicity of this U compound. Calculated U speciation indicated that Ca2UO2(CO3)3 and (UO2)2CO3(OH)3- complexes predominated under our experimental conditions. The immobilization/lethality was observed after 9 h of exposure for both U compounds. However, only UN caused a significant decrease (≈40%) in the acetylcholinesterase (AChE) activity when compared with control. In order to observe preliminary toxicity effects, we evaluated oxidative stress parameters, such as catalase (CAT) activity, TBARS formation, radical species (RS) generation and cell membrane injury and/or apoptosis (CMI). In this study, we demonstrate that U compounds caused a significant decrease in CAT activity. Similarly, we also observed that UN increased TBARS levels in AS at concentrations 5 times lower than AU (10 µM and 50 µM, respectively). Furthermore, RS generation and CMI were enhanced only on AS treated with UN. Overall, the effects observed here were remarkably significant in AS exposed to UN when compared with AU. In this study, we showed different profiles of toxicity for both U compounds, contributing significantly to the current and scarce understanding of the aquatic ecotoxicity of this heavy metal.

Zika virus-induced brain malformations in chicken embryos.

BACKGROUND: Zika virus (ZIKV) was confirmed to be related to microcephaly in 2016. However, there is still a need for understanding the embryonic morphological changes induced by ZIKV and when they occur. Here, chicken embryos were chosen as experimental model of ZIKV to evaluate virus-associated morphological alterations that might take place during embryonic development. METHODS: A screening with different viral doses was conducted in embryos at HH Stage 10-12 (E1.5) as well as a follow up of the first 5 days postinfection (dpi) was performed to observe the main morphologic changes post ZIKV infection. RESULTS: ZIKV exposed embryos presented a higher prevalence of mortality and defects such as brain malformation when compared to controls. Moreover, we observed that the phenotypes become more evident at 4dpi, when the viral load quantification reaches a peak. CONCLUSIONS: We found that ZIKV exposed embryos presented a high prevalence of mortality and central nervous system (CNS) abnormalities in a dose-dependent manner. The phenotype was more evident 4 days postinfection, when the viral load quantification reached a peak.

Effect of maternal and postnatal cocoa supplementation on testicles of adult Wistar rats.

Early weaning can lead to changes in the morphology of organs in adulthood, and the consumption of functional foods during lactation and postnatal life is believed to prevent these changes. However, it is not known if early weaning affects testicular morphology and if the use of cocoa can prevent that. We studied the effects of maternal and postnatal supplementation of cocoa powder on the testicular morphology of early weaned adult rats. The animals were divided into four groups (n = 6 each), control group, cocoa control group, early weaning (EW) group, and cocoa early weaning (EWCa) group, and were analyzed for 90 d, after which they were euthanized. The animals from the EW group showed a reduction in the tubular diameter and height of the seminiferous epithelium, a decrease in epithelial surface density (Sv), and an increase in the lumen and proper tunic. However, the animals from the EWCa group showed an increase in the diameter and height of the epithelium, an increase in the epithelium Sv, and a decrease in the lumen and the proper tunic. The early weaning promotes morphological changes in the testicles; however, supplementation with cocoa powder can preserve the testicular histoarchitecture.

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.

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.

The Brazilian-Portuguese version of the Sleep Hygiene Index (SHI): validity, reliability and association with depressive symptoms and sleep-related outcomes.

OBJECTIVE: To translate the Sleep Hygiene Index (SHI) to Brazilian Portuguese, to describe its psychometric properties and to show its association with sleep quality, daytime sleepiness, risk for sleep apnea and depressive symptoms. METHODS: Thirty subjects participated in the cultural adaptation and the item clarity evaluation. Twenty subjects answered the instrument in three different time-points for test-retest reliability. Eighty adult workers completed the SHI, the Pittsburgh Sleep Quality Index (PSQI), the Epworth Sleepiness Scale (ESS), the Beck Depression Inventory (BDI) and the STOP-BANG (S-B). RESULTS: SHI shows an acceptable internal consistency (Cronbach's α=0.75), as well as a high reproducibility (intraclass correlation=0.972, p<0.01). The three final factors of confirmatory factor analysis extract an average of 48.22% of the total sample variance. Worse sleep hygiene (higher SHI score) correlated with poor sleep quality (r=0.398, p<0.001), excessive daytime sleepiness (r=0.406, p<0.001) and depressive symptoms (r=0.324, p=0.003). No correlations with S-B were found. CONCLUSIONS: SHI presents satisfactory-to-optimal psychometric properties. This instrument is useful for treatment planning and management of sleep hygiene practices. Thus, it represents a reliable way of assessing sleep hygiene quantitatively in both research and clinical settings.

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.

Effects of the dutasteride and sildenafil association in the penis of a benign prostatic hyperplasia animal model.

OBJECTIVE: The aim of this study was to investigate whether concomitant treatment of dutasteride and sildenafil could prevent structural changes in the penis of a BPH rodent model. METHODS: Thirty-two adult male rats were divided into the following groups: Ctrl, untreated control rats; BPH, untreated spontaneously hypertensive rats (SHRs); BPH + D, SHRs treated with dutasteride; and BPH + DS, SHRs treated with dutasteride and sildenafil. All treatments were performed during 40 days, following which the penises were collected for histomorphometrical analysis. The results were compared via one-way ANOVA with Bonferroni's post-test, considering p values <.05 as significant. RESULTS: The smooth muscle density decreased by 28.6% and 21.4% in BPH + D and BPH + DS, respectively, when compared to the BPH group. The sinusoid space density reduced by 32.2% in BPH, when compared to the Ctrl group; this density was also reduced by 22.6% in BPH + D, when compared to the BPH group. The density of the elastic fibers increased 51.6% and 65.6% in BPH + D and BPH + DS, when compared to the BPH group. CONCLUSION: Treatment with dutasteride promoted morphological changes in the corpus cavernous of this BPH model. Concomitant treatment with sildenafil did not prevent the morphological changes caused by dutasteride; on the contrary, it also promoted a further increase in elastic fibers.