Metabolic and ultrastructural renal changes in adult Wistar rats fed by a cafeteria diet.

PURPOSE: To evaluate, by quantitative and qualitative methods, the glomerular ultrastructure in Wistar rats fed a cafeteria diet. METHODS: Male Wistar rats were divided into two groups at 21 days of age: control (C, n = 10) and cafeteria diet (CAF, n = 8). The animals were followed up until 5 months of age, followed by euthanasia. The blood, kidneys, and fat deposits--epididymal, retroperitoneal, and subcutaneous--were extracted and analyzed. Data were analyzed by Student's t test, and p < 0.05 was considered statistically significant. RESULTS: The cafeteria diet promoted glucose intolerance, hyperglycemia (p < 0.0001), and deposition of retroperitoneal fat (p < 0.005). Scanning electron microscopy revealed that the length of the foot process was similar in both groups. The quantitative analyses by transmission electron microscopy revealed that the cafeteria diet reduced the thickness of the glomerular basement membrane (p < 0.05). CONCLUSIONS: The intake of lipids and simple carbohydrates were found to be associated with alteration in the glomerular ultrastructure. However, more studies are needed to evaluate not only the effects of high-protein and high-fat diets on components of the glomerular filtration barrier, but also renal physiology.

Effects of chronic restraint stress in the prostate of prepubertal and adult rats.

PURPOSE: To investigate the effects of chronic stress in the prostate of prepubertal and adult rats. METHODS: Thirty-two male rats were assigned into four groups depending on the type of treatment (control or stressed) and the age at which stress was initiated (prepubertal or adult). Restraint stress stimuli were applied for six weeks. Stressed prepubertal and adult rats evaluated immediately after the last stress stimuli were named SP and SA groups, respectively. Age-matched rats were used as control groups (CP and CA). At the end of the experiment, the rats were euthanized, and prostate morphological parameters were evaluated and statistically compared. RESULTS: Application of stress stimuli to the SP group resulted in reduced body weight, but no prostate morphological modification was noted. The SA group showed reduced testosterone level and prostatic epithelium surface density, in comparison to CA group. Further, the prostatic lumen surface density was increased in adult stressed animals, in comparison to adult controls. CONCLUSIONS: The stress stimuli promoted changes in hormonal and morphological parameters in the prostate of adult stressed rats. Prepubertal stressed animals did not presented modifications of prostate morphology.

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 chronic restraint stress in the prostate of prepubertal and adult rats

Purpose: To investigate the effects of chronic stress in the prostate of prepubertal and adult rats. Methods: Thirty-two male rats were assigned into four groups depending on the type of treatment (control or stressed) and the age at which stress was initiated (prepubertal or adult). Restraint stress stimuli were applied for six weeks. Stressed prepubertal and adult rats evaluated immediately after the last stress stimuli were named SP and SA groups, respectively. Age-matched rats were used as control groups (CP and CA). At the end of the experiment, the rats were euthanized, and prostate morphological parameters were evaluated and statistically compared. Results: Application of stress stimuli to the SP group resulted in reduced body weight, but no prostate morphological modification was noted. The SA group showed reduced testosterone level and prostatic epithelium surface density, in comparison to CA group. Further, the prostatic lumen surface density was increased in adult stressed animals, in comparison to adult controls. Conclusions: The stress stimuli promoted changes in hormonal and morphological parameters in the prostate of adult stressed rats. Prepubertal stressed animals did not presented modifications of prostate morphology.

Study of Penis Structure and Development in Fetus With Prune Belly Syndrome.

OBJECTIVE: To evaluate the influence of prune belly syndrome (PBS) on the development of penises in human fetuses. METHODS: We studied 39 human fetuses (including 4 with PBS) aged 11-22 weeks post conception. We measured the length and width of the free portion and penis root and the penis total length (PTL). In 21 fetuses of the control group and in 3 with PBS we analyze the penile with histochemical and immunohistochemical methods. The total penile area, area of corpora cavernosa, area of corpus spongiosum, and thickness of tunica albuginea were measured by stereological method. Means were compared using the Kolmogorov-Smirnov and ANOVA test (P <.05). RESULTS: We did not observe difference in PTL (P = .999) when we compared the control group (4.69-29.77 mm, mean = 16.2 mm; SD = 6.34 mm) with the PBS (12.23-23 mm, mean = 16.16; SD = 4.99). The total penile area (P = .002), the area of corpora cavernosa (P = .023) and the area of corpus spongiosum (P = .004) had a significant increase when comparing the PBS with the control group. CONCLUSION: In fetuses with PBS there is no change in the length of the penis but we observed a low rate of growth in all the other histologic parameters analyzed, suggesting that PBS impacts the penile development.

Histomorphometric comparison of the corpus cavernosum of rats submitted to euthanasia with ketamine and xylazine or isoflurane.

PURPOSE: To compare the penile histoarchitecture of rats euthanized with isoflurane or with ketamine and xylazine. METHODS: Fourteen male rats were divided into two groups: ISO, with animals euthanized with isoflurane; and K+X, with animals euthanized with ketamine (150 mg/kg) associated with xylazine (15 mg/kg). Immediately after the death, the penises were dissected, fixed in a 4% buffered formalin solution, and processed for histomorphometric analysis. The surface densities (Sv) of the corpus cavernosum structures (connective tissue, smooth muscle, sinusoidal space, and elastic fibers) were evaluated using Image J software. The distribution of collagen types I and III was qualitatively assessed. Statistical analyses were performed using the Student's t test for data comparison, considering it statistically significant when p < 0.05. RESULTS: Regarding the Sv of connective tissue, smooth muscle and sinusoidal space, there were no differences between animals in both groups. On the other hand, the animals euthanized with the association of ketamine and xylazine showed the Sv of elastic fibers 24.8% higher in relation to animals euthanized with isoflurane. CONCLUSIONS: The euthanasia method affected one of the morphological parameters of the rat penises. The choice of euthanasia method must be standardized to reduce bias and to obtain reliable and reproducible results.

Does treatment with dutasteride or finasteride has impact on renal morphology? Experimental study.

PURPOSE: To investigate whether renal modifications occur following treatment with dutasteride or finasteride. METHODS: Twenty-four male rats were divided into three groups: control (that received distilled water), dutasteride (0.5 mg/kg/day), and finasteride (5 mg/kg/day) groups. All administrations were given by gavage for 40 consecutive days. After inducing euthanasia, blood was collected for urea and creatinine analyses, and both the kidneys were collected for stereological analyses of kidney morphology. RESULTS: Serum urea and creatinine levels were increased in both the finasteride and the dutasteride groups compared with those in the control group. In addition, kidney weight, kidney volume, cortical volume, glomerular volumetric density, and mean glomerular volume were reduced in both treatment groups. Finally, the number of glomeruli per kidney was reduced by 26.8% in the finasteride group and by 51.6% in the dutasteride group compared with that in the control group. CONCLUSIONS: The 5-ARIs finasteride and dutasteride promoted morphological and functional damages in rat kidneys. In addition, rats in the dutasteride group showed more severe renal modifications than those in the finasteride group.

Prostatic alterations associated to early weaning and its relation with cocoa powder supplementation. Experimental study in adult wistar rats.

Early weaning can predispose the offspring to greater risk of developing chronic diseases in adulthood. It is believed that the consumption of functional foods is able to prevent these effects. The aim of this study is to evaluate the effects of maternal and postnatal cocoa powder supplementation on body mass, metabolism, and morphology of the prostate of early weaned Wistar rats. The animals were divided into four experimental groups according to lactation time (21 or 18 days, n=6, each) as follows: control group (C), cocoa control group (CCa), early weaning group (EW), and cocoa early weaning group (EWCa). The animals were euthanized at 90 days of age. Serum biochemical analysis and prostate histomorphometric evaluation were performed. The animals supplemented with cocoa powder were heavier than their respective controls (p <0.05), although with no difference in food intake among the groups. Likewise, these same groups showed a reduction in the serum glucose in relation to C and EW groups (p <0.0001). With respect to the prostate, there was no difference in smooth muscle and lumen area densities, while the EW group had a lower epithelial height and a higher percentage of mast cells than the C group (p <0.05). On the other hand, the EWCa group managed to reverse these parameters, leveling with the controls. Early weaning resulted in hyperglycemia and important morphological changes in the prostate. In contrast, dietary supplementation with cocoa powder attenuated these effects on the metabolism and prostatic histoarchitecture, proving to be a good nutritional treatment strategy.

Anatomic study of verumontanum during endoscopic surgeries in patients with benign prostatic hyperplasia

ABSTRACT Introduction and objective: To evaluate changes in verumontanum anatomy in patients with benign prostatic hyperplasia (BPH) who used 5-alpha reductase inhibitors (5-ARIs) and to propose an anatomical classification of the verumontanum. Materials and Methods: We studied 86 patients with BPH and 7 patients without the disease (age under 40 years-old who underwent kidney or ureteral lithotripsy). Of the patients with BPH, 34 (mean age=67.26) had 5-ARIs use and 52 (mean age=62.69) did not use the drug. During surgeries, photographs of the seminal colliculus were taken and later, with the aid of software (Image J), the length (longitudinal diameter) and width (transverse diameter) of the verumontanum were measured in all patients. During the procedure, we evaluated the different types of verumontanum. For statistical analysis, the R-Project software was used. Results: In the group of patients with BPH who were taking medication (group 1), the mean measures of length and width of the verumontanum were 4.69mm and 2.94mm respectively. In the group of patients with BPH who did not use the drug (group 2), the mean diameters were 4.54mm and 3.20mm respectively. In the control group (group 3), the average length and width were 5.63mm and 4.11mm respectively. There was an increase in longitudinal and transverse measurements of the control group with an increase in body mass index (BMI) (p=0.0001 and p=0.035 respectively). In addition, there was a reduction in transverse diameter in the group of BPH using 5-ARI with increased prostate volume (p=0.010). We found five different verumontanum types: "volcano" (51.61%), "lighthouse" (24.73%), "whale tail" (12.90%), "hood" (5.38%) and "castle door" (5.38%), which we propose as an anatomical classification. Conclusion: Veromontanum has smaller measurements in patients with BPH regardless of treatment. In the control group, there was an increase in verumontanum diameters with an increase in BMI. The volcano type of verumontanum was the most frequent regardless of groups and BMI.

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.

Anatomic study of verumontanum during endoscopic surgeries in patients with benign prostatic hyperplasia.

INTRODUCTION AND OBJECTIVE: To evaluate changes in verumontanum anatomy in patients with benign prostatic hyperplasia (BPH) who used 5-alpha reductase inhibitors (5-ARIs) and to propose an anatomical classification of the verumontanum. MATERIALS AND METHODS: We studied 86 patients with BPH and 7 patients without the disease (age under 40 years-old who underwent kidney or ureteral lithotripsy). Of the patients with BPH, 34 (mean age=67.26) had 5-ARIs use and 52 (mean age=62.69) did not use the drug. During surgeries, photographs of the seminal colliculus were taken and later, with the aid of software (Image J), the length (longitudinal diameter) and width (transverse diameter) of the verumontanum were measured in all patients. During the procedure, we evaluated the different types of verumontanum. For statistical analysis, the R-Project software was used. RESULTS: In the group of patients with BPH who were taking medication (group 1), the mean measures of length and width of the verumontanum were 4.69mm and 2.94mm respectively. In the group of patients with BPH who did not use the drug (group 2), the mean diameters were 4.54mm and 3.20mm respectively. In the control group (group 3), the average length and width were 5.63mm and 4.11mm respectively. There was an increase in longitudinal and transverse measurements of the control group with an increase in body mass index (BMI) (p=0.0001 and p=0.035 respectively). In addition, there was a reduction in transverse diameter in the group of BPH using 5-ARI with increased prostate volume (p=0.010). We found five different verumontanum types: "volcano" (51.61%), "lighthouse" (24.73%), "whale tail" (12.90%), "hood" (5.38%) and "castle door" (5.38%), which we propose as an anatomical classification. CONCLUSION: Veromontanum has smaller measurements in patients with BPH regardless of treatment. In the control group, there was an increase in verumontanum diameters with an increase in BMI. The volcano type of verumontanum was the most frequent regardless of groups and BMI.

Does treatment with dutasteride or finasteride has impact on renal morphology? Experimental study

ABSTRACT Purpose: To investigate whether renal modifications occur following treatment with dutasteride or finasteride. Methods: Twenty-four male rats were divided into three groups: control (that received distilled water), dutasteride (0.5 mg/kg/day), and finasteride (5 mg/kg/day) groups. All administrations were given by gavage for 40 consecutive days. After inducing euthanasia, blood was collected for urea and creatinine analyses, and both the kidneys were collected for stereological analyses of kidney morphology. Results: Serum urea and creatinine levels were increased in both the finasteride and the dutasteride groups compared with those in the control group. In addition, kidney weight, kidney volume, cortical volume, glomerular volumetric density, and mean glomerular volume were reduced in both treatment groups. Finally, the number of glomeruli per kidney was reduced by 26.8% in the finasteride group and by 51.6% in the dutasteride group compared with that in the control group. Conclusions: The 5-ARIs finasteride and dutasteride promoted morphological and functional damages in rat kidneys. In addition, rats in the dutasteride group showed more severe renal modifications than those in the finasteride group.

Histomorphometric comparison of the corpus cavernosum of rats submitted to euthanasia with ketamine and xylazine or isoflurane

ABSTRACT Purpose: To compare the penile histoarchitecture of rats euthanized with isoflurane or with ketamine and xylazine. Methods: Fourteen male rats were divided into two groups: ISO, with animals euthanized with isoflurane; and K+X, with animals euthanized with ketamine (150 mg/kg) associated with xylazine (15 mg/kg). Immediately after the death, the penises were dissected, fixed in a 4% buffered formalin solution, and processed for histomorphometric analysis. The surface densities (Sv) of the corpus cavernosum structures (connective tissue, smooth muscle, sinusoidal space, and elastic fibers) were evaluated using Image J software. The distribution of collagen types I and III was qualitatively assessed. Statistical analyses were performed using the Student's t test for data comparison, considering it statistically significant when p < 0.05. Results: Regarding the Sv of connective tissue, smooth muscle and sinusoidal space, there were no differences between animals in both groups. On the other hand, the animals euthanized with the association of ketamine and xylazine showed the Sv of elastic fibers 24.8% higher in relation to animals euthanized with isoflurane. Conclusions: The euthanasia method affected one of the morphological parameters of the rat penises. The choice of euthanasia method must be standardized to reduce bias and to obtain reliable and reproducible results.

Fetal programming by high-fat diet promoted the decreased of the prostate in adult Wistar albino rats.

We investigated the effect of a high-fat diet on body metabolism and ventral prostate morphology in 4-months-old offspring. The mother was fed with a control (C) or a high-fat (HF) diet during gestation and lactation. At weaning, the offspring diet remained the same (C/C, n = 8; HF/HF, n = 8) or it was switched (C/HF, n = 8; HF/C, n = 9). Biometry, blood pressure (BP), glucose, lipid metabolism and ventral prostate were evaluated. Triacylglycerol of HF/C increased, and the C/HF group had decreased HDL-c levels (P = 0.0005 and P = 0.0100, respectively). All groups on the HF diet presented hyperglycemia (P = 0.0064). Serum testosterone diminished in the C/HF group (P = 0.0218). The HF diet, regardless of the period, reduced prostatic acinar area (P < 0.0001). The epithelium height was smaller in HF/C and HF/HF groups compared with C/C and C/HF (P < 0.0001), and the volume density of epithelium was lower in HF/C group compared with the C/C and C/HF (P = 0.0024). The volume density of smooth muscle cells diminished in C/HF and HF/C (P = 0.0013), and the volume density of connective tissue was reduced in HF/C and HF/HF (P < 0.0001). High-fat diet intake during prenatal and postnatal life leads to prostatic atrophy, which may impair prostate secretory activity and contractility, and thus disturb reproductive function in adulthood.

Effects of tadalafil to prevent injury on corpus cavernosum after vascular or nervous peri-prostatic bundle injury. Experimental model in rats.

PURPOSE: To evaluate the effects of tadalafil (TD) in preventing histological alterations of the corpus cavernosum caused by isolated lesions of cavernous nerve (ILCN) and artery (ILCA) in rats. METHODS: Fifty male Wistar rats were randomly assigned in five groups: G1: control; G2: bilateral ILCN; G3: bilateral ILCA; G4: ILCN+TD; G5: ILCA+TD. The cavernous bodies were submitted to histomorphometry, immunohistochemistry and biochemical analysis. RESULTS: Nerve density was significantly higher in G2 and G4 compared to control (22.62±2.84 and 19.53±3.47 vs. 15.72±1.82; respectively, p<0.05). Smooth muscle density was significantly lower in G2 and G3 in comparison to G1 (12.87±1.90 and 18.93±1.51 vs. 21.78±1.81, respectively; p<0.05). A significant decrease in the sinusoidal lumen area was observed in G2 compared to controls (5.01±1.62 vs. 9.88±3.66, respectively; p<0.05) and the blood vessel density was increased in G2 and G3 (29.32±4.13 e 20.80±2.47 vs. 10.13±2.71, p<0.05). Collagen density was higher in G3 compared to G1 (93.76±15.81 vs. 64.59±19.25; p<0.05). CONCLUSIONS: Histomorphometric alterations caused by ILCN were more intense than those produced by vascular injury, but the collagen analyses showed more fibrosis in animals with ILCA. TD was effective in preventing the majority of the alterations induced by the periprostatic bundle injury.

Expression of stromal elements of prostatic adenocarcinoma in different gleason scores.

PURPOSE: To quantify and compare the expression of stromal elements in prostate adenocarcinoma of different Gleason scores with non-tumor area (control). METHODS: We obtained 132 specimens from samples of prostate peripheral and transition zone. We analyzed the following elements of the extracellular matrix: collagen fibers, elastic system, smooth muscle fibers and blood vessels. The tumor area and non-tumor area (control) of the TMA (tissue microarray) were photographed and analyzed using the ImageJ software. RESULTS: The comparison between the tumor area and the non-tumor area showed significant differences between stromal prostate elements. There was an increase of collagen fibers in the tumor area, mainly in Gleason 7. Elastic system fibers showed similar result, also from the Gleason 7. Blood vessels showed a significant increase occurred in all analyzed groups. The muscle fibers exhibited a different behavior, with a decrease in relation to the tumor area. CONCLUSIONS: There is a significant difference between the extracellular matrix in prostate cancer compared to the non-tumor area (control) especially in Gleason 7. Important modifications of the prostatic stromal elements strongly correlate with different Gleason scores and can contribute to predict the pathological staging of prostate cancer.

Expression of stromal elements of prostatic adenocarcinoma in different gleason scores

Abstract Purpose: To quantify and compare the expression of stromal elements in prostate adenocarcinoma of different Gleason scores with non-tumor area (control). Methods: We obtained 132 specimens from samples of prostate peripheral and transition zone. We analyzed the following elements of the extracellular matrix: collagen fibers, elastic system, smooth muscle fibers and blood vessels. The tumor area and non-tumor area (control) of the TMA (tissue microarray) were photographed and analyzed using the ImageJ software. Results: The comparison between the tumor area and the non-tumor area showed significant differences between stromal prostate elements. There was an increase of collagen fibers in the tumor area, mainly in Gleason 7. Elastic system fibers showed similar result, also from the Gleason 7. Blood vessels showed a significant increase occurred in all analyzed groups. The muscle fibers exhibited a different behavior, with a decrease in relation to the tumor area. Conclusions: There is a significant difference between the extracellular matrix in prostate cancer compared to the non-tumor area (control) especially in Gleason 7. Important modifications of the prostatic stromal elements strongly correlate with different Gleason scores and can contribute to predict the pathological staging of prostate cancer.

Different sample preparation methods for glomerular morphological evaluation: to coat or not to coat? / Diferentes métodos de preparación de muestras para la evaluación morfológica glomerular ¿recubrir o no recubrir?

The renal glomerulus is coated by fenestrated endothelial cells and externally covered by specialized epithelial cells, known as podocytes. Scanning electron microscopy becomes an important and effective tool for its studies. Normally, samples destined for scanning microscopy are covered with a thin metallic layer. However, this step can be dispensed for some analyzes. We aimed to compare coated and uncoated samples for evaluation of the glomerular morphology of the Wistar rat kidney. Cortical region of the kidney of the 5month-old male Wistar rats were used. The fragments followed the routine procedure for scanning electron microscopy processing. Half of 10 fragments were coated with palladium gold and the remaining were not coated. Auriga Compact FIB - SEM scanning electron microscope was used to observe the samples. Different increases and voltages was evaluated. For the uncoated samples, when using voltages of 2 KV (or higher) a great charging was observed, impairing the use of such voltage. Thus, these samples were always observed under voltage of 0.5 KV. On the other hand, in the coated samples, the use of 2 KV was adequate. Almost as a consequence, in the coated samples, the podocyte structures were better characterized, generating better images. Inversely, in the uncoated samples, it was possible to visualize the desired structures and to detect the morphological characteristics of these. The results showed that it is possible to use kidney samples without previous coating to evaluate the glomerular morphology at the ultrastructural level, serving as a tool in the study of pathologies.

El glomérulo renal está recubierto por células endoteliales fenestradas y cubierto externamente por células epiteliales especializadas, conocidas como podocitos. La microscopía electrónica de barrido se convierte en una herramienta importante y efectiva para sus estudios. Normalmente, las muestras destinadas a microscopía de barrido se cubren con una capa metálica delgada. Sin embargo, este paso se puede dispensar para algunos análisis. El objetivo fue comparar muestras recubiertas y no recubiertas para evaluar la morfología glomerular del riñón de rata Wistar. Se utilizó la región cortical del riñón de ratas Wistar macho de 5 meses de edad. Se realizó el procedimiento de rutina para el procesamiento de microscopía electrónica de barrido. La mitad de 10 fragmentos se recubrieron con oro paladio y los restantes no se recubrieron. Se utilizó un microscopio electrónico de barrido SEM Auriga Compact FIB para observar las muestras. Se evaluaron diferentes aumentos y voltajes. Para las muestras no recubiertas, al usar voltajes de 2 KV (o más) se observó una gran carga, impidiendo el uso de dicho voltaje. Por lo tanto, estas muestras siempre se observaron a bajo voltaje de 0,5 KV. Por otro lado, en las muestras recubiertas, el uso de 2 KV fue adecuado. Como consecuencia, en las muestras recubiertas, las estructuras de los podocitos se caracterizaron mejor, generando mejores imágenes. Inversamente, en las muestras no recubiertas, fue posible visualizar las estructuras deseadas y detectar las características morfológicas de éstas. Los resultados mostraron que es posible utilizar muestras de riñón sin recubrimiento previo para evaluar la morfología glomerular a nivel ultraestructural, que sirve como una herramienta en el estudio de patologías.