ABSTRACT Background: To compare Gleason 7 (3+4) and (4+3) prostatic adenocarcinoma (PC) with different prognostic criteria through immunohistochemical analysis with anti-PSA, anti-Ki 67 and anti-AMARC antibodies. Methods: We analyzed 221 surgical specimens from patients between 40 and 86 years-old (mean=63) with PC. The immunohistochemical study was performed with anti-PSA, anti-Ki 67 and anti-AMARC. The microscopic fields were photographed with an Olympus DP70 digital camera coupled to an Olympus BX51 microscope and archived in TIFF. Proportion and intensity criteria were used to quantify the anti-PSA antibody and for the anti-Ki 67 antibody, the quantification by similarity of this antibody in breast carcinomas. Anti-AMACR protein expression was based on four scores: negative, weak, moderate and strong. The statistical analysis was performed with the Graph Pad Prism 5 program. Results: In the Gleason score 7 (3+4) we had 91.72% in pT2 and 8.27% in the pT3 group; 8.27% recurrences, of which 90.90% in the pT2 group. In the Gleason score 7 (4+3) we had 77.27% in the pT2 group and 22.72% in the pT3 group and 10.22% of relapses, of which 66.66% in the pT2 group and 33.33% in the pT3 group. In 6.81% of cases there was an increase in the anti-Ki 67 index and in 2.27% of the cases, there was an increase in the immunoexpression of anti-p53 when comparing Gleason score 7 (3+4) with Gleason score 7 (4+3). Conclusion: Our study confirmed differences in the Gleason score 7 (3+4) and Gleason score 7 (4+3) of PC when comparing prognostic criteria. Anti-Ki 67 and anti-PSA antibody immunostaining showed a positive correlation as the Gleason score 7 increased from (3+4) to (4+3).

ABSTRACT Introduction: Although nerves and vessels of the penis play important role in erection, there are few studies on their development in human fetus. Therefore, the objective of the present study is to analyze, quantitatively, in the corpora cavernosa and corpus spongiosum, the development of the nerves and vessels in the fetal penis at different gestational ages. Material and Methods: Fifty-six fresh, macroscopically normal human fetuses aged from 13 to 36 weeks post-conception (WPC) were used. Gestational age was determined by the foot length criterion. Penises were immediately fixed in 10% formalin, and routinely processed for paraffin embedding, after which tissue sections from the mid-shaft were obtained. We used immunohistochemical staining to analyze the nerves and vessels in the corpus cavernous and in the corpus spongiosum. These elements were identified and quantified as percentage by using the Image-J software. Results: The quantitative analysis showed that the percentage of nerves varied from 3.03% to 20.35% in the corpora cavernosa and from 1.89% to 23.88% in the corpus spongiosum. The linear regression analysis indicated that nerves growth (incidence) in the corpora cavernosa and corpus spongiosum correlated significantly and positively with fetal age (r2=0.9421, p<0.0001) and (r2=0.9312, p<0.0001), respectively, during the whole fetal period studied. Also, the quantitative analysis showed that the percentage of vessels varies from 2.96% to 12.86% in the corpora cavernosa and from 3.62% to 14.85% in the corpus spongiosum. The linear regression analysis indicated that vessels growth (appearance) in the corpora cavernosa and corpus spongiosum correlated significantly and positively with fetal age (r2=0.8722, p<0.0001) and (r2=0.8218, p<0.0001), respectively, during the whole fetal period studied. In addition, the linear regression analysis demonstrated a more intense growth rate of nerves in the corpus spongiosum during the 2nd trimester of gestation, when compared with nerves in the corpora cavernosa. In addition, the linear regression analysis demonstrated a more intense growth rate of vessels in the corpus spongiosum when compared with the corpora cavernosa, during the whole fetal period studied. Conclusions: In the fetal period, the human penis undergoes major developmental changes, notably in the content and distribution of nerves and vessels. We found strong correlation between nerves and vessels growth (amount) with fetal age, both in the corpora cavernosa and corpus spongiosum. There is significant greater proportional number of nerves than vessels during the whole fetal period studied. Also, nerves and vessels grow in a more intense rate than that of the corpora cavernosa and corpus spongiosum areas.

ABSTRACT 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.

ABSTRACT Purpose: To analyze the histology and distribution of abdominal testicular vessels in human fetuses Patients and Methods: We studied 19 fetuses (34 testes) ranging in age from 12 to 19 weeks post-conception. The fetuses were evaluated regarding crown-rump length (CRL), total length (TL) and body weight immediately before dissection. Each testis was dissected and embedded in paraffin, from which 5 µm thick sections were obtained and stained with Masson's trichrome and Anti-CD31 antibody to quantify the vessels. The stereological analysis was carried out with the Image Pro and Image J programs, using a grid to determine volumetric densities (Vv). Means were statistically compared using the unpaired T-test (p<0.05). Results: The fetuses presented mean weight of 222.5g, mean CRL of 15.3 cm and mean TL of 23.2 cm. All testes were in the abdominal position. The mean percentage of vessels (Vv) in the upper portion of the testis was 7.6% (4.6 to 15%) and in the lower portion the mean was 5.11% (2.3 to 9.8%), with a significant difference (p=0.0001). In the analysis between the upper portion of the right and left testes (p=0.99) and in the analysis of the lower portion of the right and left testes (p=0.83), we did not observe significant differences. Conclusion: The upper portion of the abdominal testis in human fetuses had a higher concentration of vessels than the lower portion. These results suggest that manipulation of the lower end of the testis during Fowler-Stephens surgery should be avoided in order to preserve the collateral circulation.

ABSTRACT 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. model C, C (C water n=10 n10 10 n=10) D (D 05 0 5 0. mgKgday mg Kg day T, (T 04 4 DT, (DT 10. . 10) gavage days analyses oneway one way Bonferronis Bonferroni s posttest, posttest post test, test post-test p005 p p<0.0 significant Sv, , (Sv) crosssectional cross sectional n=1 n1 1 p00 p<0. (Sv n= p0 p<0 p<

ABSTRACT 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.

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.

Abstract The increase in the number of studies related to the health of university students has attracted attention at the time of COVID-19. This initiative contributes to preparing the school environment for the return of face-to-face activities with conducts that minimize the damage caused by the pandemic. The role of each actor in the educational scenario is fundamental for us to be able to reap good fruits after this storm.

Resumo Os investimentos em sistemas de tratamento de resíduos de suínos (STRS) buscam muito mais contemplar questões de segurança, sanidade e de atendimento a requisitos da legislação ambiental do que investigar possibilidades de explorar economicamente os resíduos devidamente tratados. Este estudo analisou o retorno de investimentos de diferentes STRS sendo realizado entre os meses de outubro de 2013 a maio de 2014. Para a coleta de dados, foi elaborado um questionário eletrônico aplicado por meio de entrevistas em 37 granjas do estado de São Paulo escolhidas aleatoriamente. Para análise de viabilidade econômica dos diferentes STRS, utilizou-se a técnica de orçamento de capital (PAYBACK e TIR). As granjas foram divididas em quatro categorias: GEEBC - Geração de energia elétrica, biofertilizante e créditos de carbono; GEEBI - Geração de energia elétrica e biofertilizante; GEEEL - Geração de energia elétrica; SGPVA - Sem geração de produtos de valor agregado. O melhor resultado econômico foi observado na categoria GEEBC com tempo médio de retorno financeiro de nove anos. Observou-se que 73% dos pesquisados não exploravam o valor econômico dos resíduos tratados, mantendo seus investimentos com intuito de contemplar a legislação ambiental vigente.

Abstract The investments in wastewater treatment of swine systems (WTSS) search on safety, health and fulfillment of the requirements of environmental legislation to investigate to the possibilities of economically exploit properly treated effluent. This study analyzed the return on investment of different WTSS. Was being carried out between the months of October of 2013 to May of 2014. For data collection an electronic questionnaire applied through interviews on 37 farms in the State of São Paulo chosen randomly. To analyze the economic feasibility of different WTSS we used the technique of Capital Budget (PAYBACK and IRR). The farms were divided into four categories: GEEBC-generated electricity, biofertilizer and carbon credits; GEEBI-generated electricity and biofertilizer; GEEEL-generated electricity; NGVAP- no generating value-added products. The best economic result was observed in the category GEEBC with an average Payback time of nine years. It was observed that 73% of respondents do not exploit the economic value of the treated effluent, keeping your investments with a view to contemplate the environmental regulations.

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.

SUMMARY: 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.

RESUMEN: 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.

ABSTRACT Purpose: To evaluate if late hormonal replacement is able to recover the prostatic tissue modified by androgenic deprivation. Materials and Methods: 24 rats were assigned into a Sham group; an androgen deficient group, submitted to bilateral orchiectomy (Orch); and a group submitted to bilateral orchiectomy followed by testosterone replacement therapy (Orch+T). After 60 days from surgery blood was collected for determination of testosterone levels and the ventral prostate was collected for quantitative and qualitative microscopic analysis. The acinar epithelium height, the number of mast cells per field, and the densities of collagen fibers and acinar lumen were analyzed by stereological methods under light microscopy. The muscle fibers and types of collagen fibers were qualitatively assessed by scanning electron microscopy and polarization microscopy. Results: Hormone depletion (in group Orch) and return to normal levels (in group Orch+T) were effective as verified by serum testosterone analysis. The androgen deprivation promoted several alterations in the prostate: the acinar epithelium height diminished from 16.58±0.47 to 11.48±0.29μm; the number of mast cells per field presented increased from 0.45±0.07 to 2.83±0.25; collagen fibers density increased from 5.83±0.92 to 24.70±1.56%; and acinar lumen density decreased from 36.78±2.14 to 16.47±1.31%. Smooth muscle was also increased in Orch animals, and type I collagen fibers became more predominant in these animals. With the exception of the densities of collagen fibers and acinar lumen, in animals receiving testosterone replacement therapy all parameters became statistically similar to Sham. Collagen fibers density became lower and acinar lumen density became higher in Orch+T animals, when compared to Sham. This is the first study to demonstrate a relation between mast cells and testosterone levels in the prostate. This cells have been implicated in prostatic cancer and benign hyperplasia, although its specific role is not understood. Conclusion: Testosterone deprivation promotes major changes in the prostate of rats. The hormonal replacement therapy was effective in reversing these alterations.

Abstract Purpose: To investigate the glomerular number after different warm ischemia times. Methods: Thirty two pigs were assigned into four groups. Three groups (G10, G20, and G30) were treated with 10, 20, and 30 minutes of left renal warm ischemia. The sham group underwent the same surgery without renal ischemia. The animals were euthanized after 3 weeks, and the kidneys were collected. Right kidneys were used as controls. The kidney weight, volume, cortical-medullar ratio, glomerular volumetric density, volume-weighted mean glomerular volume, and the total number of glomeruli per kidney were obtained. Serum creatinine levels were assessed pre and postoperatively. Results: Serum creatinine levels did not differ among the groups. All parameters were similar for the sham, G10, and G20 groups upon comparison of the right and left organs. The G30 group pigs' left kidneys had lower weight, volume, and cortical-medullar ratio and 24.6% less glomeruli compared to the right kidney. A negative correlation was found between warm ischemia time and glomerular number. Conclusions: About one quarter of glomeruli was lost after 30 minutes of renal warm ischemia. No glomeruli loss was detected before 20 minutes of warm ischemia. However, progressive glomerular loss was associated with increasing warm ischemia time.

ABSTRACT PURPOSE: To evaluate the glomerular loss after arteriovenous or arterial warm ischemia in a swine model. METHODS: Twenty four pigs were divided into Group Sham (submitted to all surgical steps except the renal ischemia), Group AV (submitted to 30 minutes of warm ischemia by arteriovenous clamping of left kidney vessels), and Group A (submitted to 30 minutes of ischemia by arterial clamping). Right kidneys were used as controls. Weigh, volume, cortical volume, glomerular volumetric density (Vv[Glom]), volume-weighted glomerular volume (VWGV), and the total number of glomeruli were measured for each organ. RESULTS: Group AV showed a 24.5% reduction in its left kidney Vv[Glom] and a 25.4% reduction in the VWGV, when compared to the right kidney. Reductions were also observed when compared to kidneys of sham group. There was a reduction of 19.2% in the total number of glomeruli in AV kidneys. No difference was observed in any parameters analyzed on the left kidneys from group A. CONCLUSIONS: Renal warm ischemia of 30 minutes by arterial clamping did not caused significant glomerular damage, but arteriovenous clamping caused significant glomerular loss in a swine model. Clamping only the renal artery should be considered to minimize renal injury after partial nephrectomies.