C. Gallo

Effects of castration and hormone replacement in the urinary bladder of rats: structural, ultrastructural, and biochemical analysis.

We evaluated, by qualitative and quantitative methods, the structural alterations in the bladder wall of rats submitted to surgical castration, as well as the role of hormone replacement in reversing the possible structural alterations. Twenty-four 12-week-old male Sprague-Dawley rats were used. The animals were divided into 3 groups comprising 8 animals each and treated as follows. Members of group CONTR (control) underwent a sham operation only and were sacrificed after 2 months. Members of group ORCH (orchiectomy) underwent bilateral orchiectomy and were sacrificed after 2 months. Members of group ORCH+TEST (testosterone) underwent orchiectomy, received testosterone replacement after 1 month, and were sacrificed 1 month later. We performed a qualitative and quantitative analysis of collagen by light microscopy, scanning electron microscopy, biochemistry, and a histomorphometric analysis of smooth muscle and elastic fibers in the 3 groups. The results showed a significant decrease in absolute values of elastic fibers in the castrated group. The histomorphometric analysis of epithelial height did not show differences among the groups. There was no statistical difference in quantitative analysis of collagen, either by histomorphometry or by biochemistry. Also, there was no difference in the smooth muscle cells. However, the qualitative analysis revealed differences in collagen (castrated group) when compared with controls and with rats submitted to hormone replacement. Hormone replacement with testosterone was able to revert the alterations observed. The findings suggest that hormone replacement, even when instituted at a late stage, is effective in reversing the bladder wall alterations produced by secondary hypogonadism.

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.

Leptin and leptin receptor expressions in prostate tumors may predict disease aggressiveness

PURPOSE The aim of this study was to evaluate the expression of leptin and its receptor in histological sections of prostate tumors, and their association with prognostic factors. METHODS A total of 532 surgical specimens from prostate cancer were studied. After histopathological diagnosis, the samples were included in tissue microarrays containing cores from tumor and non-tumor (benign prostatic hyperplasia) areas. These were immunostained with anti-leptin and anti-leptin-receptor antibodies. Objective and subjective analyses were performed. Students-t-test and ANOVA were used to compare mean values, and linear regression was used to evaluate the correlation between histological results and prognostic indicators. RESULTS Leptin receptor expression was reduced in tumors with a positive surgical margin, urethral margin involvement, and seminal vesicles invasion. Further, there was a negative correlation between the expression of leptin receptor in tumor areas and the sum of prognostic factors, suggesting that leptin receptor may predict the aggressiveness of disease. CONCLUSION Our findings suggest that leptin receptor expression is a potential prognostic factor for PCa. Further investigation is needed to support the use of leptin receptor as a novel biomarker, although leptin itself does not seem to predict the aggressiveness of prostate cancer.

Modifications of Erectile Tissue Components in the Penis during the Fetal Period

Background The penile erectile tissue has a complex microscopic anatomy with important functions in the mechanism of penile erection. The knowledge of such structures is necessary for understanding the normal physiology of the adult penis. Therefore, it is important to know the changes of these penile structures during fetal development. This study aims to analyze the development of the main components of the erectile tissue, such as collagen, smooth muscle fibers and elastic system fibers, in human fetuses. Methodology/Principal Findings We studied the penises of 56 human fetuses aged 13 to 36 weeks post-conception (WPC). We used histochemical and immunohistochemical staining, as well as morphometric techniques to analyze the collagen, smooth muscle fibers and elastic system fibers in the corpus cavernosum and in the corpus spongiosum. These elements were identified and quantified as percentage by using the Image J software (NIH, Bethesda, USA). From 13 to 36 WPC, in the corpus cavernosum, the amount of collagen, smooth muscle fibers and elastic system fibers varied from 19.88% to 36.60%, from 4.39% to 29.76% and from 1.91% to 8.92%, respectively. In the corpus spongiosum, the amount of collagen, smooth muscle fibers and elastic system fibers varied from 34.65% to 45.89%, from 0.60% to 11.90% and from 3.22% to 11.93%, respectively. Conclusions We found strong correlation between the elements analyzed with fetal age, both in corpus cavernosum and corpus spongiosum. The growth rate of these elements was more intense during the second trimester (13 to 24 WPC) of gestation, both in corpus cavernosum and in corpus spongiosum. There is greater proportional amount of collagen in the corpus spongiosum than in corpus cavernosum during all fetal period. In the corpus spongiosum, there is about four times more collagen than smooth muscle fibers and elastic system fibers, during all fetal period studied.

Ultrastructural Analysis of the Foreskin in Patients With True Phimosis Treated or Not Treated With Topical Betamethasone and Hyaluronidase Ointment

OBJECTIVE To evaluate, by using scanning electron microscopy (SEM), the possible alterations of the foreskin connective tissue in patients with true phimosis submitted to topical treatment with betamethasone and hyaluronidase ointment. MATERIALS AND METHODS We studied 15 patients (mean 5.3 years old) submitted or not to topical application of betamethasone 0.2% and hyaluronidase cream. For qualitative analysis of the connective tissue, we studied 5 samples from each foreskin, with 2 mm length. The samples were submitted to fixation for SEM by immersing tissue fragments in a modified Karnovsky solution for 48 hours at 4°C. The obtained acellular preparations were then processed for high-vacuum SEM, and observations were performed on a LEO 435 (Zeiss, Oberkochen, Germany) scanning electron microscope with an acceleration voltage of 15 to 20 kV. RESULTS Parents of 7 children did not agree with the clinical treatment and opted for circumcision directly. These patients served as the control group (nontreated). Eight patients submitted to topical treatment could not expose the glans and were referred for circumcision. In SEM, with a magnification of 5000×, we observed important differences in the organization of the collagen and elastic system fibers when comparing treated and nontreated patients with betamethasone and hyaluronidase. Treated patients presented a different organization of collagen with a clear decrease in the elastic system fibers. CONCLUSION The treatment of phimosis with betamethasone + hyaluronidase showed changes in the structure of the foreskin with a decrease of elastic system fibers, which is characteristic of the healing processes.

Structural changes of dilated pelvic ureters in adults

PURPOSE To perform a quantitative assessment of different types of pelvic ureter tissues with chronic dilation in adults, using stereological methods. METHODS We analyzed fragments of dilated pelvic ureters obtained from 6 patients aged between 35 and 67 years (mean, 46 years) who underwent ureteral reimplantation surgery for different reasons. The obstruction duration ranged from 27 to 180 days (mean, 93 days). The control group included fragments of normal pelvic ureters obtained during nephrectomy in 7 kidney transplant donors (age: range, 25-51 years; mean, 36 years). The volumetric density of collagen in the ureter, elastic fibers, and smooth muscle fibers was assessed. RESULTS The volumetric density (Vv) of collagen showed no significant difference between the two groups (control: 45.3 ± 6.1; dilated: 40.8 ± 6.9; P = 0.23). A statistically significant increase in Vv of elastic fibers was observed in the dilated ureters (control: 18.4 ± 1.2; dilated: 24.6 ± 5.4; P = 0.03). A statistically significant increase in the Vv of smooth muscle fibers was observed in the dilated ureter (control: 42.0 ± 6.0; dilated: 56.2 ± 6.1; P = 0.001). CONCLUSION The chronically dilated pelvic ureters had significantly more elastin and smooth muscle contents than the controls.

Development of the Penis during the Human Fetal Period (13 to 36 Weeks after Conception)

PURPOSE We analyzed the development of the area of the penis and erectile structures (corpora cavernosa and corpus spongiosum) and the thickness of the tunica albuginea during the fetal period (13 to 36 weeks after conception) in humans to establish normative patterns of growth. MATERIALS AND METHODS We studied 56 male human fetuses at 13 to 36 weeks after conception. We used histochemical and morphometric techniques to analyze the parameters of total penile area, area of corpora cavernosa, area of corpus spongiosum, and thickness of tunica albuginea in the dorsal and ventral regions using ImageJ software (National Institutes of Health, Bethesda, Maryland). RESULTS Between 13 and 36 weeks after conception the area of the penis varies from 0.95 to 24.25 mm2. The area of the corpora cavernosa varies from 0.28 to 9.12 mm2, and the area of the corpus spongiosum varies from 0.14 to 3.99 mm2. The thickness of the tunica albuginea varies from 0.029 to 0.296 mm in the dorsal region and from 0.014 to 0.113 mm in the ventral region of the corpora cavernosa. CONCLUSIONS We found a strong correlation between the total penile area, corpora cavernosa and corpus spongiosum with fetal age (weeks following conception). The growth rate was more intense during the second trimester (13 to 24 weeks of gestation) compared to the third trimester (25 to 36 weeks). Tunica albuginea thickness also was strongly correlated with fetal age and this structure was thicker in the dorsal vs ventral region.

Histological and quantitative analyzes of the stromal and acinar components of normal human prostate zones

McNeal divided the human prostate into three major anatomical areas: the peripheral zone (PZ), the central zone (CZ), and the transition zone (TZ). Each of these areas is biologically and histologically distinct. The PZ and TZ have clinical significance and are associated with prostate cancer (PC) and benign prostatic hyperplasia (BPH), respectively. Therefore, the objective of the present study was to quantitatively and qualitatively analyze the parenchymal and stromal components that constitute the different prostate zones.

PD49-12 VITAMIN-D RESTRICTION DURING GESTATION IN WISTAR RATS CAUSES ALTERATION IN VESSELS AND NERVES OF THE PENIS IN THE OFFSPRING

and neuronal nitric oxide synthase (nNOS), tyrosine hydroxylase (TH) and TUNEL assay expression were measured 72h. Gene expression of nNOS, TH, beta-tubulin, Schwann cells (glial fibrillary acidic protein; GFAP), markers of nerve injury (activating transcription factor 3; ATF3) and regeneration (growth associated protein 43, GAP43) was also measured in irradiated MPGs (n1⁄46/grp). RESULTS: In dissociated MPG neuronal cultures, there was an early increase in neuron length, while branching and nNOS positive neurons decreased (p<0.05). Early radiation caused a 2-fold increase in apoptotic neurons (p<0.05). However, the gene expression of neuronal markers beta-tubulin, nNOS, TH and markers of injury and repair (ATF3, GAP43) were all unchanged. There was a marked increase in the gene expression of Schwann cell marker GFAP 2 weeks post-RT (p<0.001). At 10 weeks post-RT, there was a 20% decrease in neuron length, decreased neuron branching, and 20-30% less nNOS and TH positive neurons (p<0.05). Additionally, there was a 2.5 fold increase in the number of TUNEL positive apoptotic neurons (p<0.05). Gene expression of nNOS, TH, GAP43 and ATF3 were all decreased (p<0.05) while GFAP remained considerably elevated (p<0.005). Interestingly, erectile function was not impaired at either time point following radiation. CONCLUSIONS: This is the first study to characterize the health and regeneration potential of MPG neurons following RT. While this model lead to minimal changes in erectile function, neuronal injury was apparent early post-RT and persisted by increasing over time. The nerves are very susceptible to apoptosis and damage from prostatic RT and additional research is necessary to develop radioprotective strategies for exposed benign periprostatic tissues.

Effects of Vitamin D Restricted Diet Administered during Perinatal and Postnatal Periods on the Penis of Wistar Rats

Vitamin D deficiency is common in pregnant women and infants. The present study aimed to investigate the effects of vitamin D restricted diet on the Wistar rats offspring penis morphology. Mother rats received either standard diet (SC) or vitamin D restricted (VitD) diet. At birth, offspring were divided into SC/SC (from SC mothers, fed with SC diet) and VitD/VitD (from VitD mothers, fed with VitD diet). After euthanasia the penises were processed for histomorphometric analysis. The VitD/VitD offspring displayed metabolic changes and reduction in the cross-sectional area of the penis, corpus cavernosum, tunica albuginea, and increased area of the corpus spongiosum. The connective tissue, smooth muscle, and cell proliferation percentages were greater in the corpus cavernosum and corpus spongiosum in the VitD/VitD offspring. The percentages of sinusoidal spaces and elastic fibers in the corpus cavernosum decreased. The elastic fibers in the tunica albuginea of the corpus spongiosum in the VitD/VitD offspring were reduced. Vitamin D restriction during perinatal and postnatal periods induced metabolic and structural changes and represented important risk factors for erectile dysfunction in the penis of the adult offspring. These findings suggest that vitamin D is an important micronutrient in maintaining the cytoarchitecture of the penis.