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.

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

BACKGROUND: 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. METHODS: We assessed 19 samples from each prostate zone. The samples were obtained from necropsies of young people between 18 and 32 years of age with intact urogenital tracts. The samples were fixed in 4% buffered formalin and processed for paraffin embedding. Sections with a thickness of five micrometres were obtained from each sample. The sections were stained using histochemical and immunohistochemical techniques to identify the acinar and stromal components of each zone. Photomicrographs were obtained for morphometric analysis using an algorithm based on color segmentation. Data were analyzed using one-way analysis of variance (ANOVA) with the Bonferroni post-test. Differences with P < 0.05 were regarded as statistically significant. RESULTS: Collagen fibres were more numerous in the TZ (+40.26%; P = 0.0230) than in the PZ. Muscle fibres were also more numerous in the TZ (+47.05%; P = 0.0120) than in the PZ. Elastic system fibres in the TZ significantly differed from those in the PZ (+84.61%; P = 0.0012) and the CZ (+61.66%; P = 0.0074). Similarly, nerves in the PZ (-42.86%; P = 0.0107) significantly differed from nerves in the CZ. Epithelial height was lower in the TZ than in the PZ (-30.17%; P = 0.0034) and the CZ (-25.01%; P = 0.0330). CONCLUSION: Our objective, quantitative data regarding the various elements that constitute the normal prostate stroma allowed us to reveal differences among prostate zones. This study established patterns for normal parameters and may be used for posterior comparisons in histopathological analysis.

The prostate after castration and hormone replacement in a rat model: structural and ultrastructural analysis

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.

The prostate after castration and hormone replacement in a rat model: structural and ultrastructural analysis.

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.

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.

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

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

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. Student's-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.

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. Student's-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. .

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.

Nutritional supplementation with L-arginine prevents pelvic radiation-induced changes in morphology, density, and regulating factors of blood vessels in the wall of rat bladder.

PURPOSE: To determine whether L-arginine has protective effects against radiation-induced alterations in the morphology and regulatory factors of vesical blood vessels in rats. METHODS: Male rats aged 3-4 months were divided into groups of 10 animals each: (a) controls, consisting of non-treated animals; (b) radiated-only rats; and (c) radiated rats receiving L-arginine supplementation. Radiation was in one session of 10 Gy and was aimed at the pelvic-abdominal region. L-arginine was administered once a day (0.65 g/kg body weight), starting 7 days before radiation and continuing until killing on the 16th day after radiation. The density, relative area, and wall thickness of blood vessels were measured in the vesical lamina propria using histological methods, and the expression of vascular endothelial growth factor (VEGF) and fibroblast growth factors (FGF) in the bladder wall was assessed by RT-PCR. RESULTS: Compared with controls, radiation alone decreased the density and relative area of blood vessels by 32 % (p < 0.01) and 25 % (p < 0.05), respectively, and reduced the arterial wall thickness by 42 % (p < 0.004). VEGF and FGF mRNA levels after radiation were diminished by 67 % (p < 0.002) and 56 % (p < 0.04), respectively. The radiated animals supplemented with L-arginine were not significantly different from controls. CONCLUSIONS: Pelvic radiation leads to significant vesical modifications, as in the morphology of blood vessels and in VEGF and FGF expression. All these changes, however, were prevented by L-arginine treatment. These results emphasize, therefore, the potential use of this amino acid as a radioprotective drug.

Effects of castration and late hormonal replacement in the structure of rat corpora cavernosa.

The objective of the study was to evaluate, through quantitative methods, the structural alterations in the corpora cavernosa of rats submitted to orchiectomy as well as the role of late hormone replacement in overturning the possible structural alterations. Twenty-five male rats were assigned into 5 groups with 5 animals each and treated as follows: ORCHIEC-1 = submitted to orchiectomy and sacrificed after 1 month; C-1 = control group sacrificed after 1 month; ORCHIEC-2 = submitted to orchiectomy and sacrificed after 2 months; C-2 = control group sacrificed after 2 months; and T = submitted to orchiectomy, underwent testosterone replacement with testosterone undecanoate (100 mg/kg) after 1 month, and sacrificed after 1 month of hormonal replacement. Smooth muscle, collagen, and elastic system fibers of penile corpora cavernosa were quantified. There was a significant decrease in the absolute values of smooth muscle, sinusoidal space, and total area of corpora cavernosa after 2 months in the castrated group when compared with controls. Overall, regarding density, no significant differences were observed among the groups. The hormonal replacement with testosterone was able to reverse the alterations observed. The method used for this research allowed demonstrating that absolute values are reliable to quantify the structural alterations of corpora cavernosa structures. The results suggest that hormonal replacement, even when instituted at a late stage, is effective in reversing the corpora cavernosa's structural alterations produced by castration.

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.