Abstract: The present study deals with the stereological analysis of the changes in the seminiferous epithelium of rat testis, mediated either by cadmium alone or by cadmium plus zinc, chronically administered at low oral doses. Stereological estimates of both labeling indices of proliferative cell nuclear antigen and TUNEL-stained nuclei of
spermatogonia, and of ubiquitin, ubiquitin carboxyl-terminal hydrolase L1, caspase-3, and metallothionein cytoplasmic immunoreactivities from seminiferous epithelium were performed in rats non-treated (controls), CdCl2-exposed animals, and CdCl2 plus ZnCl2 -treated rats. The following conclusions can be drawn: a) The intake of low, oral doses of cadmium chloride over a long period of time induces quantitative changes in apoptosis of the seminiferous epithelium of the rats, without noticeable morphologic or proliferative alterations. b) The increase of mono-ubiquitin levels mediated by cadmium is caused by over expression of ubiquitin carboxyl-terminal hydrolase L1. c) Zinc exposure was able to decrease ubiquitin carboxyl-terminal hydrolase L1 and ubiquitin, but not sufficiently to reverse the apoptotic rate of the spermatogonia at the control level. d) It seems that metallothionein is not induced by the cadmium treatment alone. However, the results indicate that either cadmium in combination with zinc or zinc itself induces this enzyme.
Abstract: The amount of neurons of periprostatic accessory ganglia in pre- and peripubertal rats was studied to
ascertain whether the development of these autonomic ganglia is androgen-dependent. Stereological
estimates of the volumes and number of neurons immunoreactive to protein gene product 9.5 (PGP 9.5),
neuropeptide Y (NPY), and vasoactive intestinal polypeptide (VIP) were carried out. Immunostaining of
androgen receptors (AR) in the ganglia was also performed. The ganglionic neurons from the two groups
studied were immunoreactive to PGP 9.5, NPY, and VIP. Almost all the neurons were immunostained for
AR. The ganglionic volume showed a significant increase in peripubertal prostate in comparison with the
prepubertal gland. No significant changeswere observed with respect to the absolute number of neurons
immunoreactive to all the antigens. The neuronal volume was significantly increased in peripubertal rats
in comparison with prepubertal animals. These findings led us to the following conclusions: There is no
evidence of neurogenesis during pubertal development in the periprostatic accessory ganglia of the rat.
The increase of ganglionic volume in puberty is due to the growth in neuronal volume. There were no
differences between the sizes of NPY and VIP neurons in pubertal periprostatic accessory ganglia. The
development of periprostatic vegetative neurons is androgen-dependent.
Abstract: Portal hypertension induces a splanchnic and systemic low-grade inflammatory response that could induce the expression of three phenotypes, named ischemia-reperfusion, leukocytic, and angiogenic phenotypes.During the splanchnic expression of these phenotypes, interstitial edema, increased lymph flow, and lymphangiogenesis are produced in the gastrointestinal tract. Associated liver disease increases intestinal bacterial translocation, splanchnic lymph flow, and induces ascites and hepatorenal syndrome. Extrahepatic cholestasis in the rat allows to study the worsening of the portal hypertensive syndrome when associated with
chronic liver disease. The splanchnic interstitium, the mesenteric lymphatics, and the peritoneal mesothelium seem to create an inflammatory pathway that could have a key pathophysiological relevance in the production of the portal hypertension syndrome complications. The hypothetical comparison between the ascitic and the amniotic fluids allows for translational investigation. From a phylogenetic point of view, the ancestral mechanisms for amniotic fluid production were essential for animal survival out of the aquatic environment. However, their hypothetical appearance in the cirrhotic patient is considered pathological since ultimately they lead to ascites development. But, the adult human being would take advantage of the potential beneficial effects of this âamniotic-like fluidâ to manage the interstitial fluids without adverse effects when chronic liver disease aggravates.
Abstract: It is currently believed that portal hypertension induces an inflammatory response in
which mast cells may be involved. The aim of this study was to verify the
involvement of the intestinal submucosal and mesenteric lymph node mast cells in
the splanchnic inflammatory response related to portal hypertension. Mast cell
infiltration in the intestine (duodenum, jejunum, ileum, caecum and distal colon)
and in the mesenteric lymph node complex (MLC) was measured using a stereological
method in sham-operated rats (SO; n ¼ 12), in two experimental models of portal
hypertension, chronic (triple partial portal vein ligation, TPVL; n ¼ 12) and transient
(microsurgical partial hepatectomy; n ¼ 12) and in rats in which the MLC was
resected (n ¼ 12). The small and large bowel submucosal infiltration increases in
MLC-resected rats (p ¼ 0.0001), in TPVL rats (p ¼ 0.0001) and in rats with partial
hepatectomy (p ¼ 0.0001). An extensive mast cell infiltration in the MLC
(p ¼ 0.0001) was found in TPVL rats and in rats with partial hepatectomy
(347.40745.25 and 351.92799.28/mm3, respectively) in relation to sham-operated
rats (135.27730.28/mm3). We conclude that mast cells could be involved in the
splanchnic alterations developed in the surgical experimental models of portal
hypertension studied.
Abstract: Abnormal angiogenesis is a critical feature of many diseases, including cancers and their precursors. Although
the association between prostate carcinogenesis and changes in microvascular architecture is well known, these changes are not well-documented from a quantitative point of view. The present work deals with stereological estimates of the number of quiescent and proliferative endothelial cells, and microvessel length in normal and prostate cancer tissues. Unbiased stereological measurements of numerical densities of proliferating cell nuclear antigen immunostained cells, nonproliferating endothelial cells, caspase 3 immunoreactive endothelial cells, and relative length (length density) of microvessels, were performed in control and cancer specimens. There were no changes in either proliferation or apoptosis in carcinoma endothelial cells. A decrease of endothelial cell density, together with an increase of microvessel length density,
were detected in prostate cancer specimens. Therefore, the following conclusions can be drawn: a) The increase of angiogenetic activity in prostate carcinogenesis leads to an increment of the microvascular length; b) The amount of endothelial cells per vascular length decreases in prostate cancer; c) There is no decrease of endothelial apoptosis in cancer microvessels. d) The increase of the length density of microvessels in prostate cancer is not directly associated to an enhancement of the endothelial proliferation; and e) The blood supply of epithelium was similar in both cancerous and normal prostate.
Abstract: The interactions between steroid and nonsteroid hormones in the prostate are of special interest during the growth phase of the gland. The purpose of this work is to study the influence of prolactin (PL), with or without androgenic blockade, on epithelial cells from peripubertal rat ventral prostate. Twenty male peripubertal Sprague-Dawley rats were grouped as controls, or treated with cyproterone acetate (CA), CA plus PL (CA-PL), or PL. The total number (N total) of epithelial cells, and their labeling indices to proliferative cell nuclear antigen (LI PCNA), apoptosis (LI apoptosis) and androgen receptors (LI AR) were measured. CA and PL treatment significantly decrease the N total, but the LI PCNA was unchanged. We have observed a greater LI apoptosis in pharmacologically castrated animals without PL than in the rats with androgenic blockade with PL. The LI AR does not change with CA treatment in the ventral region, but the PL significantly increases it. Androgenic blockade and PL decrease the number of epithelial cells from the ventral prostate. These changes are not attributable to the decrease of cell proliferation, rather to the increase of epithelial apoptosis. The increase of cells expressing AR after treatment with PL might be attributed to the decrease of testosterone secretion caused by the hyperprolactinemia. PL does not modulate the size of the ventral prostate in prepubertal rats. Anat Rec, 2009. (c) 2009 Wiley-Liss, Inc.
Abstract: This work deals with the quantification of serotonin-immunoreactive prostate neuroendocrine cells (NECs) in rats exposed to prolactin in normal, cyproterone acetate-exposed, and bromocriptine-exposed animals to establish the possible influence of prolactin with or without androgenic blockade on this cell population. Thirty male peripubertal Sprague-Dawley rats were grouped as controls (CT) and those treated with cyproterone acetate (CA), cyproterone acetate plus prolactin, cyproterone acetate plus bromocriptine, prolactin (PL), and bromocriptine (BC). The volume of ductal epithelium (Vep) and total number (NSER) of the NECs serotonin-immunoreactive were measured. NECs were detected in the periurethral ducts. Compared to CT, Vep was increased in PL and BC and NSER was decreased in CA and increased in the prolactin or bromocriptine groups. The androgenic blockade decreases NSER in rat prostate; PL induces in normal and cyproterone acetate-treated rats the increase of NSER; and BC exerts a local effect over the prostate similar to that described for PL.
Abstract: Eosinophilic esophagitis (EE) is an emerging disease caused by dense infiltration of the esophageal epithelium by eosinophilic leucocytes. It is originated from local hypersensitivity to food or airborne allergens. Although the physiopathologic mechanisms of the illness have not been fully discovered, EE is a loss of immunologic tolerance by the esophagus, meaning that it should be considered as an active immunologic organ. In our study, we investigated the immunologic capacity of the epithelium using immunohistochemistry and stereology, to determine the cellular density of eosinophils, T and B lymphocytes, Langerhans cells, mast cells, and cells manufacturing immunoglobulin E in endoscopic biopsies of patients with EE (taken before and after topical treatment with fluticasone propionate) compared with normal individuals and patients suffering from gastroesophageal reflux disease (GERD). We have observed that the density of eosinophils in EE is 300 times greater than in normal conditions and it is only in this disease where eosinophils show signs of activation and degranulation (positivity to major basic protein immunostaining). The number of T intraepithelial lymphocytes also significantly rose in EE, compared with other entities, where CD8 cells were predominant. However, the human esophagus is deficient in B lymphocytes and we only found intraepithelial plasma cells that excreted immunoglobulin E in EE. Under normal conditions mast cells exist in the thickness of the epithelium that are slightly higher in GERD and multiply in density by 17 in EE. Langerhans cells did not show any significant variation in density under the different tested conditions. After topical treatment with steroids, the density of the different cell components fell to similar levels to GERD. Using our study, we can conclude that the human esophagus may contribute to the development of local immunologic responses as it contains all the necessary cell components. EE represents growth of this esophageal capacity and its pathogeny could respond to mixed cellular and humoral mechanisms.
Abstract: This work was directed to evaluate immunoexpression of markers for apoptosis, resistance to apoptosis, and cell proliferation, as well as estimates of nuclear size in ventral prostate of rats treated with cadmium chloride and cadmium+zinc chloride because a possible protective effect of zinc has been postulated. The following variables were studied: volume fraction (VF) of Bcl-2 immunostaining, percentage of cells immunoreactive to proliferating cell nuclear antigen (LIPCNA) and p53 (LIp53), numerical density of caspase-3 immunoreactive cells (NV caspase-3), and estimates of volume-weighted mean nuclear volume (upsilonV). The LIPCNA and VF of Bcl-2 were significantly increased in the treated animals. The dysplasias (independent of their origin) showed a significant increase of the LIp53, NV caspase-3, and upsilonV in comparison with normal acini from treated and control animals. It can be concluded that cell proliferation is enhanced in long-term cadmium-exposed rats, and exposure to zinc combined with cadmium had no effect on any of the variables studied when comparing with normal acini. The increase of nuclear upsilonV could indicate a more aggressive behavior for pretumoral lesions.
Abstract: It has been shown that there are sugars in corpora amylacea, but little attention has been focused on the expression of glycoconjugates in corpora amylacea of normal and hyperplastic prostatic glands. The present study characterizes and compares the expression of glycoconjugates in corpora amylacea of normal and hyperplastic prostatic glands of elderly men by using alcian blue (AB) stain and lectin histochemistry. Corpora amylacea were larger and more numerous in hyperplastic glands compared to normal glands. The stain with AB revealed the presence of sulfated and carboxyl components in corpora amylacea. In hyperplastic prostatic glands the sulfur and acid contents of corpora amylacea were increased. Lectin affinities of corpora amylacea from normal prostatic glands demonstrated the presence of fucose, mannose, sialic acid, N-acetyl galactosamine and N-acetyl glucosamine residues. In the hyperplastic glands the lectin binding pattern of corpora amylacea was qualitatively similar to normal glands, but an increase in GalNAc, sialic acid, mannose and fucose residues was observed. Normal prostatic glands showed a weak to moderate content of mannose residues, and in contrast a strong GNA and Con-A staining was observed in hyperplastic glands. MAA and SNA affinities indicated that the content of sialic acid residues was higher in hyperplastic glands compared with normal prostatic glands. Also NAcGal residues were increased in hyperplastic glands. Luminal secretion, secretory cells and apical border of epithelium showed a similar although more intense Lectin-binding pattern as compared with corpora amylacea both in normal and hyperplastic prostatic glands. Lectin histochemistry shows that the glycoconjugates expressed in the glandular epithelium are similar to those found in corpora amylacea both in normal and hyperplastic glands. In addition, in hyperplastic glands, where the corpora amylacea are higher in size and more numerous, the reaction to lectins is more intense especially with mannose and sialic acid residues. The results suggest that corpora amylacea are originated at least in part from prostatic secretion.
Abstract: BACKGROUND: Because most of the characteristics of the portal hypertensive enteropathy can be explained on the basis of increased levels of mast cell mediators, the purpose of the present paper was to study mast cell splanchnic infiltration. METHODS: Duodenum, jejunum, ileum and mesenteric lymph node complex infiltration by mast cells was assayed by a stereological technique in control rats (group I; n = 5) and in an experimental model of portal hypertension (the portal vein-stenosed rat, group II; n = 5) at 6 weeks after operation. RESULTS: Intestinal and mesenteric lymph node complex infiltration by mast cells increased in the animals with partial portal vein ligation. The mast cell density progressively increased distally along the small bowel. The mast cell increase in the mesenteric lymph node complex in portal vein-stenosed rats was greater than in the duodenum (P = 0.001), jejunum (P = 0.006) and ileum. CONCLUSION: The rise of mast cells density in the small bowel and mesenteric lymph node complex in rats with partial portal vein ligation suggests that these cells are involved in the etiopathogenesis of experimental portal prehepatic hypertensive enteropathy.
Abstract: BACKGROUND: This study was directed to evaluate the immunoexpression of markers for cell proliferation, apoptosis, nuclear androgen receptors, and Glutathione-S-Transferase P1 (GSTP1), in preneoplastic changes induced by cadmium chloride (Cd) and cadmium plus zinc chloride (Cd + Zn) in rat prostate. METHODS: The following parameters were calculated in ventral prostate of normal rats and rats that received Cd or Cd + Zn in drinking water during 24 months: numerical densities of columnar, basal, and GSTP1 immunoreactive epithelial cells; percentages of cells immunoreactive to: PCNA, (LI(PCNA)), androgen receptors (LI(AR)), and of apoptotic cells. RESULTS: The LI(PCNA) was significantly increased in the animals exposed to Cd + Zn, whereas the numerical densities of both columnar (N(V) columnar cells), and GSTP1 immunoreactive (N(V) GSTP1+) cells were significantly increased in the animals treated with metals in comparison with the controls. No significant differences between the two sources of dysplasias (Cd and Cd + Zn) respecting to LI(PCNA), N(V) columnar cells, and N(V) GSTP1+ were observed. The two types of dysplasias considered together showed a significant increase for the N(V) basal, N(V) columnar, and N(V) GSTP1+ cells in comparison with normal acini of treated and controls. The percentage of apoptotic nuclei did not show significant differences among the three groups studied. CONCLUSIONS: (1) The zinc has little influence in the development of the dysplastic changes of the rat prostate mediated by cadmium. (2) The decrease of apoptosis has little influence in the development of dysplasia. (3) GSTP1 could play a role in the response to the oxidative stress in the dysplastic changes caused by cadmium.
Abstract: This work was undertaken to study prostate innervation during the postnatal development of rats. It deals with the quantification of nervous fibers throughout all the regions of the rat prostate during the postnatal development using a general marker for nervous tissue, protein gene product 9.5, and 2 neuropeptides (NPY and VIP). Forty male Wistar rats (prepubertals, pubertals, young, and aged adults) were studied for immunohistochemistry of protein gene product (PGP 9.5), neuropeptide Y (NPY), and vasoactive intestinal polypeptide (VIP). They were also evaluated for length density of nerve fibers (L(V) PGP 9.5, L(V) NPY, L(V) VIP). Nerve fibers immunoreactive to the 3 antigens studied were detected in all the groups and in all the prostate zones. Periductal L(V) NPY evidenced a significant increase in the pubertal group, maintained throughout adult life. Periductal L(V) VIP showed a significant increase in young adults. The length densities of VIP and NPY fibers were significantly higher in periductal and ampular locations in comparison with dorsal and ventral sites. It can be concluded that the relative amount of nerve fibers in rat prostate, detected by PGP 9.5, does not change during postnatal development. There were significant changes in NPY and VIP fibers, showing an increase in periurethral ducts at puberty. The abundance of peptidergic innervation around the excretory ducts is related to their contractility. The development of innervation of periurethral ducts is regulated by androgens.
Abstract: In this study, we examined the age-related changes on morphometric parameters and ultrastructure of seminiferous tubules, and on the expression of extracellular matrix proteins in lamina propria of Syrian hamsters. A significant decrease in the percentage of normal tubules and an increase in the percentage of hypospermatogenic and arrested maturation tubules was observed with aging. Aged animals showed a decrease in tubular diameter, tubular lumen, seminiferous epithelium volume and total tubular volume. However, the total length of seminiferous tubules was significantly increased with aging. The most important ultrastructural changes with aging were the thickening of the lamina propria, the presence of diverse abnormalities in the spermiogenesis process, degeneration of germ cells, and vacuolization and flattening of Sertoli cells showing abundant lipofucsin droplets and residual bodies. Laminin immunoreactivity was found along the lamina propria of seminiferous tubules both in young and aged animals. Fibronectin immunoreactivity was found along the lamina propria and blood vessels. Both laminin and fibronectin total volume of immunostaining per testis was increased in aged hamsters. In conclusion, the age-related changes in seminiferous tubules of hamster include: a decrease in tubular width and an increase in tubular length; widening of the lamina propria caused by a more extensive connective matrix between the peritubular cells and the basal membrane; and a strong disarrangement of the seminiferous epithelium, including germ cell degeneration and important alterations in both spermiogenesis and Sertoli cell structure.
Abstract: The cellular mechanisms implicated in the atrophy of seminiferous epithelium in ageing are currently under debate, although recent reports suggest that apoptosis may be the primary mechanism implicated in aged germ cell loss. Other investigators have suggested that changes in spermatogonial proliferation are also involved. In the present work, the changes in proliferation and apoptosis in the seminiferous epithelium of aged (24 months) Syrian hamsters were examined in concert and compared with those in young (6 months) animals. Proliferation of germ cells was studied by bromodeoxyuridine labelling and apoptosis was assessed by transmission electron microscopy and in situ TUNEL labelling. Aged animals showed a significant decrease in the numbers of total and proliferating spermatogonia plus preleptotene spermatocytes per unit volume and per testis and in the proliferative index (24.8 +/- 1.6%) compared with young animals (30.8 +/- 1.2%) (P < 0.05). The number of apoptotic spermatogonia plus spermatocytes per unit volume and the apoptotic index were significantly higher in aged animals (1.51 +/- 0.23% v. 0.77 +/- 0.04%; P < 0.05). Apoptosis was confirmed by morphological characteristics: condensation of the chromatin and nuclear fragmentation. In aged hamsters, tubular degeneration could be classified into several categories, showing an increase of apoptotic cells in tubular cross-sections characterized by maturation arrest in comparison with all other types. Spermatogonial proliferation was also diminished as seen in tubular cross-sections showing hypospermatogenesis, sloughing off of germ cells and maturation arrest. The results obtained in the present study suggest that the decrease in the proliferation of spermatogonia and the increase in apoptosis constitute two consecutive mechanisms correlated with the ageing of the seminiferous epithelium.
Abstract: BACKGROUND: This work was undertaken to study the prostate neuroendocrine cells (PNEC) during the post-natal development of rats. METHODS: Forty male Wistar rats (pre-pubertals, pubertals, young, and aged adults) were used for immunohistochemistry of chromogranin A (cgA), serotonin (SER), and protein gene product 9.5 (PGP9.5). They were also evaluated for numerical cell density (NV SER) and PNEC number per prostate (N SER). Five additional young adult rats were used for a RT-PCR study (mRNA cgA detection). RESULTS: Weak immunoreactivity to cgA was observed in pubertal rats. No PNEC immunostained to PGP 9.5 was observed. Cells expressing SER were detected in all the groups exclusively located in periurethral ducts. The NV SER increased significantly in pubertal animals. In aged animals, it decreased to levels observed in pre-pubertal rats. The N SER increased significantly from pre-pubertal to young adults, decreasing in aged adults. There was weak production of cgA mRNA, with more expression in the dorsal prostate. CONCLUSIONS: PNEC differ in rats when compared to humans: they are weakly immunopositive to cgA, do not express PGP 9.5, only show immunoreactivity to SER, and do not appear in acini. The changes in the amount of rat PNEC during the post-natal development suggest an androgenic influx. PNEC might regulate the contractility of periurethral ducts.
Abstract: In the hamster, male reproductive quiescence is accomplished via testicular atrophy and the germinal epithelium is regressed to spermatogonia and spermatocytes after 8-14 weeks of short photoperiods. However, the cellular mechanisms involved in this process have not been elucidated. As it is suggested that the regulation of seasonal testicular activity is characterized by coordinated shifts in the relationships between mitosis, meiosis and apoptosis, the changes in the proliferative and apoptotic activity in the seminiferous epithelium of photoinhibited Syrian hamster were examined and compared with those maintained in natural photoperiod. The proliferative activity was studied using BrdU immunostaining, and germ cell apoptosis was assessed by in situ TUNEL labelling and transmission electron microscopy. A significant increase in the rate of apoptosis (percentage of TUNEL-positive spermatogonia + spermatocytes) was observed in photoinhibited animals (2.84 +/- 0.16) compared with those exposed to natural photoperiod (0.77 +/- 0.03, p < 0.05). The majority of apoptotic germ cells were spermatocytes and in some occasions spermatogonia. Germ cell apoptosis was confirmed by morphological characteristics: condensation of the chromatin and nuclear fragmentation. The rate of proliferation (percentage of BrdU-positive spermatogonia + preleptotene spermatocytes) was significantly higher in photoinhibited hamsters (42.7 +/- 2.6) compared with animals exposed to natural photoperiod (31.1 +/- 1.6, p < 0.05). After the exposure to a short photoperiod the apoptotic index positively correlated with the proliferative index (r = 0.8150, p < 0.05). In conclusion, the seminiferous epithelium of photoinhibited Syrian hamsters is characterized by an increased rate of apoptosis associated to an enhanced rate of proliferation.
Abstract: This work was addressed to study correlations between histopathology of breast malignancy and variations in depth, intensity, multiplicity, and simultaneous capacitance-conductance features of electrical impedance scanning (EIS), in cases presenting mammographic findings. The EIS was performed in 74 patients. The entrance criterion was the presence of either suspicious or dubious mammography. The EIS evaluation was unblinded to mammographic data. The histologic findings of patients eligible for biopsy (Bi-rads 4, 5, and Bi-rads 0 after re-evaluation) were correlated to EIS and mammography. Depth localization of lesion, intensity, multiplicity, and capacitance-conductance features of the EIS signals were evaluated. There was association between histopathological diagnosis and EIS results. The presence of multiplicity of EIS signal, or simultaneous capacitance-conductance signals, was significantly ( p<0.05) more frequent in cases with either suspicious mammography or malignant biopsy than those dubious for mammography or with benign biopsy. There was no significant relationship between depth and intensity of EIS signal. In 15 (20%) cases there was discordance among mammography, EIS, and histology (controversial cases). Six of these 15 (40%) were perimenopausal women. Benign proliferating lesion was diagnosed in 6 of 15 (40%) controversial cases. It is concluded that the multiplicity of EIS spots and simultaneousness of capacitance and conductance signals were attributed to malignancy. Detection of false-positive EIS results was common in perimenopausal patients (40%). Patients with benign proliferating lesions presented also false positivity to EIS. Mammography and EIS had similar rates of false-positive findings in this study.
Abstract: Neuroendocrine cells may play a role in both normal and pathologic conditions of the human prostate. It may be interesting to investigate 1) whether there are significant amounts of neuroendocrine cells in human adult normal prostate, and 2) whether the distribution of these cells shows regional differences. This study estimates both absolute and relative amounts of neuroendocrine cells immunostained for two neuronal markers (chromogranin A and protein gene product 9.5) and for serotonin in the three regions of human prostate, transition zone, central zone, and peripheral zone, using unbiased stereologic measurements. There was observed a predominance of neuroendocrine cells in the transition zone of the normal prostate. The neuroendocrine cells of this region may play a role in the genesis of benign prostate hyperplasia. The significant presence of neuroendocrine cells secreting neuropeptides in peripheral zone could be correlated with the induction of androgen-independent growth in prostate carcinogenesis. The wolffian origin attributed to the central zone can explain its poor population of neuroendocrine cells.
Abstract: BACKGROUND: Cadmium chloride is an environmental toxic that might be implicated in human prostate carcinogenesis. The study was directed: 1) to evaluate the immunoexpression of markers for cell proliferation, apoptosis, and resistance to apoptosis, and 2) to estimate the size of premalignant cell population in the preneoplastic changes induced in ventral prostates of rats treated with cadmium chloride administered in drinking water. METHODS: The following parameters were calculated in the ventral prostatic lobe of normal rats and rats that received cadmium in drinking water during 18 months: total volume, epithelial volume, total number of epithelial cells, numerical density of epithelial cells, percentage of cells that immunostained to the proliferating cell nuclear antigen (PCNA), percentage of apoptotic cells (evaluated by a DNA fragmentation method), and absolute volume and volume fraction of immunostaining to bcl-2. RESULTS: The percentage of PCNA immunoreactive nuclei, the bcl-2 expression, and the numerical density of epithelial cells were significantly (P < 0.05) increased in the dysplastic prostatic acini of treated rats in comparison with the normal acini of treated rats and control animals. The percentage of apoptotic nuclei from ventral dysplastic acini was significantly (P < 0.05) decreased in comparison with that of normal acini. A negative correlation between proliferation and apoptosis was found in dysplastic lesions. CONCLUSIONS: Prostate epithelial dysplasia induced in rats by cadmium presents an increased proliferative activity and high expression of bcl-2 protein, as was described in human prostate intraepithelial neoplasia. However, the rate of apoptosis in rat dysplasia was importantly decreased, in contrast to that observed in some human preneoplastic changes. This decrease might be related to the increase of bcl-2 expression.
Abstract: Immunohistochemical and semiquantitative study of TNF-alpha, its receptors types 1 (TNFR1) and 2 (TNFR2), cell proliferation (Ki-67 nuclear antigen), and apoptosis (Tunel method) was carried out in human prostates, in normal healthy conditions, as well as in benign prostatic hyperplasia (BPH) and prostatic carcinoma (PC). Cell proliferation was higher in BPH than in normal prostates, and even higher in PC, mainly in neoformations showing a microglandular pattern. The apoptotic index was similar in BPH and normal prostates, and increased significantly in PC with independence of the pattern. In BPH, immunoreaction to TNF-alpha decreased as compared with that of normal prostates, while immunoreactions to both TNF-alpha receptors increased. This suggests a feedback downregulation of the factor, and that the low TNF-alpha activity in BPH are compensated by the increased amount of receptors. In PC, immunoreaction to TNF-alpha and its two receptors increased markedly, suggesting that the TNF-induced effects are also increased. Contrarily to cell proliferation immunoexpression, PC reaction to TNFR2 was stronger in the papillar pattern than in the micrograndular pattern, and this suggests an inverse correlation between TNFR2 expression and cell proliferation.
Abstract: This study was designed to investigate (a) the presence of protein gene product 9.5 (PGP 9.5), ubiquitin, and neuropeptide Y (NPY) in the neuroendocrine and secretory epithelium of the human normal prostate and its secretions, and (b) the changes in immunoreactivity to these proteins in men with benign prostatic hyperplasia. Western blotting and light microscopic immunohistochemistry techniques were used and the numerical density of immunoreactive neuroendocrine cells, and the volume fractions of immunostained secretory epithelium were evaluated. Western blotting revealed the presence of the three antigens in both tissue homogenates and prostate secretion. Some neuroendocrine cells immunoreacted to PGP 9.5 and NPY in all the prostate regions of control specimens. Ubiquitin immunoreactivity was detected in the nuclei from both basal cells and secretory epithelial cells. The cytoplasm of the secretory cells and the glandular lumen also showed immunostaining for the three proteins. The numerical densities of both PGP 9.5 and NPY neuroendocrine cells were lower in hyperplasia than in controls. No differences in the volume fraction occupied by epithelial immunostaining to both proteins was found between hyperplastic and control prostates. We concluded that (a) PGP 9.5 and NPY, but not ubiquitin, are common antigens in both neuroendocrine and secretory prostate cells, (b) the three immunoreactive proteins contribute to the prostate secretions, and (c) the secretion of ubiquitin is markedly diminished in the hyperplastic epithelium.(J Histochem Cytochem 48:1121-1130, 2000)
Abstract: OBJECTIVE: To estimate cytologic volume-weighted mean nuclear volume and correlate it with other prognostic factors, such as tumor diameter and cytologic grading in relation to nodal infiltration. STUDY DESIGN: The relationships between nodal status and nuclear VV, tumor diameter and cytologic grading, according to the modified Black nuclear grading system, were analyzed on fine needle aspirates of 49 cases of breast cancer by univariate and multivariate logistic regression. RESULTS: Volume-weighted mean nuclear volume (nuclear VV) estimated on fine needle aspiration smears showed a significant correlation with grade of tumor differentiation. CONCLUSION: Stereologic evaluation of nuclear size by nuclear VV is an objective method for the cytologic grading of ductal carcinoma of the breast and has independent prognostic value in relation to nodal status higher than those of tumor diameter and cytologic grade.
Abstract: To investigate 1) the correlation between the proliferative activity of epididymal epithelium plus myoid cells and the increase in the number of these cells and 2) the role of the basal epithelial cells in the renewal of epididymal epithelium, a quantitative evaluation of the proliferation of both epithelial cells and periductal myoid cells in the different epididymal regions (caput, corpus, and cauda) has been carried out during postnatal development of the rat by immunohistochemical evaluation of BrdU-labeling indices. These data were correlated with cell numbers and counted by the optical dissector method. The presence of bcl-2 protein was immunohistochemically detected and evaluated. No significant differences in BrdU indices were observed among epididymal regions in any stage studied. Cell proliferation decreased from the prepubertal period to adulthood in both epithelial and myoid cells in the three regions of the epididymis, suggesting a close relationship between epithelial and mesenchymal components. The numbers of both cell types were significantly higher in the caput than in the corpus and cauda in all stages studied, suggesting functional differences between regions. A negative linear correlation between proliferative activity and cell numbers was noted that might be related to regulation of the cell population size. Basal cells showed a lower proliferation rate than principal cells, but most of the immunoreactive bcl-2 protein, in pubertal and adult epididymides, was observed in basal cells. Therefore, these cells might comprise a low-proliferating and apoptosis-resistant population.
Abstract: Colorectal cancer has been studied from the point of view of the relationships between several variables, including proliferative activity and p53 protein expression. However, stereologic evaluation of nuclear size has not been thoroughly described. In the present study, measurements of the volume-weighted mean nuclear volume (nuclear Vv), have been performed in well, moderately and poorly differentiated colorectal adenocarcinomas and correlated with the Dukes' stage and other factors such as proliferative activity and p53 protein immunoreactivity. Although the mean values of nuclear Vv were higher in poorly differentiated cancers or Dukes' C&D stage than in well and moderately differentiated tumors or Dukes' A&B stage, these differences were not significant. However, the variability of nuclear size in colorectal cancers was more relevant than the mean values of nuclear Vv with respect to their invasive classification. The carcinomas which extended beyond the serosa (Dukes' stage C&D) had higher biologic variation than those grouped as Dukes' stage A&B. The results of the present study also indicate that nuclear size in colorectal cancers has a positive correlation with both proliferative activity and p53 protein expression. The relationship between nuclear Vv and proliferative activity emphasizes the possible prognostic relevance of this stereologic estimate of nuclear size in colorectal adenocarcinoma.
Abstract: The distribution of protein gene product 9.5 (PGP) and ubiquitin immunoreactivities in the ductuli efferentes, ductus epididymidis, and ductus deferens of humans was studied by Western blot analyses and light and electron microscopic immunocytochemistry. PGP immunoreactivity was intense in the ductuli efferentes and weak in the ductus epididymidis and ductus deferens, while ubiquitin immunoreactivity was intense in the ductuli efferentes and ductus epididymidis and very weak in the ductus deferens. In the ductuli efferentes epithelium, PGP immunolabeling was observed in the cytoplasm of principal cells, whereas ubiquitin immunoreactivity was found in the nucleus and cytoplasm of principal cells and ciliated cells. In the ductus epididymidis epithelium, only scattered cells (mitochondria-rich cells) showed PGP immunoreaction in their cytoplasm, whereas ubiquitin immunostaining was detected in the nucleus and cytoplasm of most epithelial cells, except for the cauda, where ubiquitin immunolabeling was observed only in the nuclei. The ductus deferens showed no immunostaining for PGP, and only nuclear immunoreactivity to ubiquitin. The ultrastructural localization of PGP immunoreactivity was in the apical cytosol and microvilli. In addition to these locations, ubiquitin immunoreactivity was also found in the nucleus of all cell types and cilia of ciliated cells. Although the distribution of PGP and ubiquitin immunoreactivities in humans differs from that reported in rats, it seems that PGP and ubiquitinated proteins are secreted into the epididymal lumen in both species.
Abstract: The distribution of protein gene product 9.5 (PGP) and ubiquitin in the spermatozoa and epithelial cells in the different regions of the rat ductus epididymidis (proximal caput, distal caput, corpus and cauda) was studied by Western blotting analyses and electron microscopical immunogold labelling. Western blotting analyses showed that the PGP immunoreactive band was very intense in the caput and cauda epididymidis and almost irrelevant in the corpus, while the ubiquitin immunoreactive band was intense in the distal caput and cauda. No ubiquitin immunoreactive band was observed in the proximal caput and only a very weak band was seen in the corpus. The results of electron microscopical immunogold labelling varied from one epididymal region to another. The proximal caput epididymidis presented immunoreaction to PGP in the rough endoplasmic reticulum, cytosol, mitochondria and microvilli of most principal cells, and in the cytosol, rough endoplasmic reticulum and mitochondria of most basal cells. No ubiquitin immunoreaction was observed in this epididymal region. In the distal caput epididymidis, PGP immunoreactivity was detected in some principal and basal cells in the same intracellular locations as described in the proximal caput. In this region, ubiquitin immunoreactivity appears in the apical cytosol and mitochondria of principal cells. The corpus epididymidis showed no immunoreaction to PGP or ubiquitin. In the cauda epididymidis, immunostaining to PGP was observed in most clear cells and in isolated principal cells. The intracellular location of PGP in both cell types was the cytosol, mitochondria and microvilli. Ubiquitin immunoreactivity was detected in the perinuclear cytosol and mitochondria-but not in the digestive vacuoles-of some clear cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Abstract: Changes in the testicular peritubular lamina propria in rats treated for 1-11 weeks with intra-scrotal injections of epinephrine were studied by quantitative immunohistochemical methods. In control testes, BrdU-labelled nuclei (proliferating cells) were observed only in spermatogonia and some primary spermatocytes, whereas testes from epinephrine-treated rats showed BrdU labelling in some of the spermatogonia and in peritubular cells. Immunostaining for transforming growth factor beta 1 (TGF-beta 1) was present in germ cells, Sertoli cells and Leydig cells; vimentin immunostaining was found mainly in Sertoli cells; desmin immunostaining was found in the peritubular cells, and immunostaining for type IV collagen, laminin and fibronectin was found in the extracellular matrix of the lamina propria. The volume densities of seminiferous tubules (including seminiferous epithelium, lamina propria and tubular lumen) that immunostained for TGF-beta 1, vimentin, laminin, desmin or fibronectin were calculated. All of these parameters increased significantly in testes from epinephrine-treated animals during the course of the experiment, except for desmin immunostaining which showed no significant change in volume density. Since total seminiferous tubule volume decreased markedly in the testes of treated rats during the experiment, the transformation of relative values for immunostaining into absolute volumes per testis revealed a significant increase in TGF-beta 1 immunostaining, no significant change in vimentin immunostaining, and a significant decrease in desmin immunostaining during the time of the study. The absolute volume occupied by laminin and fibronectin immunostaining decreased from the 3rd to the 8th weeks of treatment, and increased from the 8th to the 11th weeks. These changes, associated with germ cell depletion and tubular fibrosis, suggest that tubular ischaemic atrophy caused by epinephrine alters the peritubular myoid cells, which change immunophenotype and increase their secretion of the extracellular matrix components producing tubular fibrosis. The mechanism of this alteration may involve direct effects on the peritubular cells or the changes may be secondary to germ cell and/or Sertoli cell lesions.
Abstract: The proliferation of peritubular myoid cells in the testes of rats treated for 1-11 weeks with intra-scrotal injections of epinephrine was investigated using immunohistochemistry and quantitative histology. The percentage of peritubular cells that were immunopositive for proliferating cell nuclear antigen (PCNA) or that were labelled with 5'-bromodeoxyuridine (BrdU) in the S-phase of the cell cycle, were calculated in control and treated rats after 1,3,5,8 and 11 weeks of treatment. In addition, the change in the number of peritubular cells per testis was calculated using two different stereological methods. The possible correlation between the changes observed using the two proliferation indices (PCNA immunoreaction and labelling of BrdU) in peritubular myoid cells was evaluated by regression analysis. The results of the study indicate that both proliferation indices increased in peritubular cells between the third and the eighth weeks of treatment, and that this increase was correlated with an increase in the number of these cells. From weeks 8-11 of treatment, both proliferation indices decreased and the same occurred with the number of peritubular cells. We hypothesize that proliferation of the peritubular cells occurs in order to increase their secretion of extracellular matrix components leading to enlargement of the lamina propria of the seminiferous tubule.
Abstract: A marked depletion of neuropeptide-immunoreactive nerves, a consequence of the nerve damage which is commonly found in leprosy, has been reported in peripheral tissues of leprosy patients and of a leprosy animal model. The aim of this study was to investigate peripheral reinnervation following a denatured autologous muscle graft in an animal model of leprosy nerve damage. Possible reinnervation of the foot-pad skin was studied by immunohistochemistry using antisera to the neuronal marker protein gene product 9.5 (PGP), the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal peptide (VIP), and the C-flanking peptide of neuropeptide Y (CPON). The extent of the reinnervation process was assessed by image analysis quantification at different time points. At 8 weeks after muscle grafting, there were small numbers of immunoreactive nerves (p < 0.05). At 12, 16, and 20 weeks postoperatively there was a gradual increase in all immunostaining. At 20 weeks, no significant difference was found for PGP-, CGRP-, and SP-immunoreactive nerves in the epidermal and subepidermal layers compared to control (contralateral) tissue. In experimental tissue the recovery of immunoreactive nerves around sweat glands took longer (up to 12 weeks) than in other skin compartments, but after that time the recovery was rapid and at 20 weeks no difference was measured for VIP-immunoreactive nerves in comparison with controls. Around blood vessels, the recovery of CGRP- and CPON-immunoreactive fibers was slow, and at 20 weeks a difference with control samples (p < 0.01) was noted. In the same area, there was no significant difference for PGP immunoreactivity between controls and tissues at 20 weeks. In contrast, the immunoreactive nerve bundles in the dermis showed a faster recovery than nerves in other skin areas, with amounts similar to controls at 20 weeks. The significant recovery of immunoreactive nerves, in particular of those containing sensory neuropeptide, is consistent with the described functional recovery.
Abstract: A immunocytochemical study for detection of proliferating cell nuclear antigen (PCNA) in order to quantify the number of PCNA-positive spermatogonia, and cytophotometric determination of spermatogonial DNA were performed in cryptorchid and control testes. The number of PCNA-positive spermatogonia, and the average DNA content of spermatogonia in the cryptorchid testes were altered from first years of age. These precocious spermatogonial alterations suggest that the early surgical testicular descent doesn't prevent lesions of germ cells.
Abstract: Nerve damage, resembling that caused by Mycobacterium leprae in man, was created by the injection of cobalt-irradiated M. leprae organisms into the tibial nerve of guinea-pigs. Assessment of nerve damage was made by clinical, electrophysiological and morphometric means at intervals up to 13 weeks after injection. Quantitative immunohistochemical analysis of neuropeptide-containing fibres in the skin of the foot was also carried out. Significant nerve damage occurred 3 weeks after injection of M. leprae organisms. Motor and sensory functional loss peaked at 5 weeks after injection, and there was a significant decrease of peptide-immunoreactive nerves in all skin compartments. The nerve damage was self-limiting and functional recovery had occurred by 13 weeks. The model shows many of the features found in the nerve damage of treated leprosy patients.
Abstract: A quantitative immunohistochemical study was performed of the distribution of protein gene product 9.5 (PGP, a soluble protein localized in neurons and neuroendocrine cells as well as in some non-nervous cells) and ubiquitin along the rat epididymis. In the ductuli efferents, PGP immunoreaction was observed in the whole cytoplasm of some columnar cells; a smaller number of columnar cells showed ubiquitin immunoreactivity with limited apical and basal cytoplasmic localization. In the proximal caput epididymidis, the whole cytoplasm of all columnar cells showed PGP immunoreactivity, ubiquitin immunostaining was negative in this region. In the middle and distal caput epididymidis and the distal cauda, the apical cytoplasm of some columnar cells and the whole cytoplasm of some basal cells showed immunoreactivity to PGP. In these regions, immunoreactivity to ubiquitin was positive in the supranuclear cytoplasm of some columnar cells but not in the basal cells. No immunoreactivity to PGP or ubiquitin was detected in the corpus epididymis and the proximal cauda. Double immunostaining revealed that all the epididymal ubiquitin immunoreactive cells were also PGP immunoreactive, whereas most PGP immunoreactive cells did not immunoreact to ubiquitin. In ubiquitin-PGP immunoreactive cells, the site of the PGP immunoreaction differed from that of the ubiquitin immunoreaction. PGP-ubiquitin immunoreactive cells also seemed to be immunoreactive to anti-AE1/AE3 keratin antibodies. The spermatozoal heads were immunoreactive to PGP antibodies in the epididymal regions from proximal caput to distal cauda but not in the ductuli efferents. The findings suggest that non-ubiquitinated PGP immunoreactive proteins are secreted in the epididymis, mainly in the proximal caput, and attach to spermatozoa.
Abstract: The DNA content of spermatogonia was studied by cytophotometric quantification in the testes of cryptorchid children and adults, as well as in age-matched control males. In most cases, the average DNA content of spermatogonia was significantly increased in the cryptorchid testes of children with uni- or bilateral cryptorchidism, as well as in the contralateral scrotal testes of children with unilateral cryptorchidism. In the group of adult men the average DNA content of spermatogonia in the testes was even more increased than in children. There were not significant differences between 4 and 14 years of age, between unilateral and bilateral cryptorchidism, or between cryptorchid testes and contralateral normally descended testes. The DNA content of spermatogonia in the surgically descended testes of 3 children who were re-biopsied 3-4 years after orchidopexy was similar before and after orchidopexy. These findings suggest that the spermatogonia of many cryptorchid males bear a congenital lesion.
Abstract: Proliferating cell nuclear antigen (PCNA) is an auxiliary protein to DNA polymerase delta necessary for tissue cellular proliferation. The colon releases several peptides or hormones which are probably related to intestinal proliferation. Colonic resection determines adaptive changes in the remnant bowel. In the present study, proliferative changes after colectomy were studied by means of the murine monoclonal PC10 antibody. A control group (n = 10 rats) and a 75% proximal colon resection group (n = 10 rats) were studied. 14 days after resection, jejunal, ileal and colon samples were taken and assayed for PCNA. Relationship between immunostained nuclei and the total number of nuclei was determined. The three intestinal segments showed statistically significant increases (p < 0.001) in the number of immunostained nuclei. PCNA proliferative index was greater in the remnant large bowel.
Abstract: The ultrastructure and immunostaining with antibodies against actin, desmin, and vimentin were studied in the peritubular myofibroblasts of testes from normal men and men with Klinefelter' syndrome (KS). The seminiferous tubules were classified into five types (a-e), related to the progressive degree of sclerosis measured as thickening of the lamina propria. In control testes, only types a and b tubules were present, whereas the testes from men with KS showed types b, c, d, and e tubules. The ultrastructural study revealed abundant microfilament bundles with electron-dense bodies in the cell periphery of the myofibroblasts in a and b tubules. In c tubules, the microfilament bundles of the myofibroblasts were lacking in electron-dense bodies. Myofibroblasts in tubules d and e showed scanty microfilament bundles. Immunostaining of peritubular myofibroblasts with anti-actin antibodies was intense in tubule types a-c and scanty in types d and e. Immunostaining of myofibroblasts with anti-desmin antibodies was intense in tubule types a and b, and negative in types c-e. Immunostaining with anti-vimentin antibodies was weak in tubule types a-c and intense in types d and e. Quantitative study revealed that with the progression of sclerosis, the number and volume per cross-sectioned tubule of actin-containing cells and, mainly, desmin-containing cells decrease while the number and volume of vimentin-containing cells increase.
Abstract: In order to compare the histology of the ampulla of the ductus deferens with that of the other segments of the duct in man, the seminal vesicle and the adjacent 13-15 cm of the ductus deferens were obtained during cystectomy from 15 adult men, and were processed for light and electron microscopy. Each ductus deferens specimen was divided into 3 segments: segment A or initial segment (the most proximal to the testis) showing a smooth outer surface and, on section, a uniform lumen and absence of mucosal invaginations; segment B (1.5-4 cm) showing a smooth outer surface and, on section, small cavities in the mucosa; and segment C or ampulla (3-4 cm), which was easily recognisable because of the cerebriform pattern on its outer surface. Segment A showed the usual histological pattern reported in studies of the human ductus deferens. Segment B consisted of mucosa, muscularis mucosae, submucosa, muscular coat and adventitia. The epithelial lining formed multiple branched invaginations in the lamina propria and submucosa giving rise to glandular structures. The lumen of the duct and the glands were lined by the same cell types: (1) basal cells; (2) mitochondrion-rich cells; and (3) columnar cells with the ultrastructural features of glycoprotein-secreting cells. The latter cells could be classified into 3 subtypes suggesting different stages of development: (a) with abundant mitochondria; (b) with abundant rough endoplasmic reticulum; and (c) with abundant secretory granules. Segment C or the ampulla showed the same histology as segment B except for the presence of many diverticula in the ampulla.(ABSTRACT TRUNCATED AT 250 WORDS)
Abstract: Ultrastructural and some immunophenotypic features of the peritubular myoid cells of testes from normal men and from men with varicocele were studied. The seminiferous tubules were classified into five types (a-e), related to the progressive degree of sclerosis measured as thickening of the lamina propria. In normal testes only type a and b tubules were found, whereas the testes from men with varicocele showed type b-e tubules. Myoid cells in tubule types a and b showed slender cytoplasmic projections with abundant, parallel arranged microfilament bundles and electron-dense bodies. In c tubules, the myoid cells showed the same ultrastructure. The myoid cells of tubules with advanced (type d) or complete (type e) sclerosis showed irregularly outlined nuclei, scant microfilament bundles and absence of electron-dense bodies. Immunostaining of myoid cells with anti-actin antibodies was intense in types a-c tubules and scant in types d and e. Immunostaining with anti-desmin antibodies was intense in tubules types a-d, but the immunoreactive cells in types c and d tubules were irregularly shaped and distributed and were scanty in tubule type e. Immunostaining with anti-vimentin antibodies was weak in types a-c tubules and intense in types d and e tubules. Quantitative studies revealed that, with the progression of sclerosis, the numbers of both actin- and desmin-immunoreactive cells per cross-sectioned tubule, and the surface area occupied by the immunostained portion of these cells, decreases while the number of vimentin-immunoreactive cells and their immunostained surface area increases.
Abstract: 125I-glucagon binding sites have been detected and quantified on the intestinal smooth muscle cells of the dog by means of ultrastructural autoradiographic methods. These binding sites are located mainly in the plasmalema. The present findings established a morphological correlation with the physiological data concerning the spasmolytic function of the glucagon on the intestinal wall.
Abstract: Among the therapeutic alternatives to orthotopic liver transplantation, hepatocyte transplantation (HT) offers the best potential in a number of liver diseases, mainly inborn errors of metabolism. Nevertheless, HT presents several inconveniences such as the scarce knowledge of the functionality of the transplanted hepatocytes, which has given rise to controversy about the specificity or unspecificity of the transplant, and the lack of a suitable system for preserving the cells. This study was designed to test a system for cryopreserving hepatocytes and to assess their functionality over prolonged periods after their ectopic transplantation. A medium and a freezing schedule which are reproducible and yield elevated viability have been used, and a number of hepatospecific parameters have been assessed: the activity of ornithine carbamoyltransferase--an enzyme of primary importance in the urea cycle--lipogenesis, gluconeogenesis, glucose-6-phosphatase and cytochrome oxidase activities, the presence of albumin--as an index of plasma protein synthesis--and IDA uptake and metabolism, showing the UDP-glucuronyl transferase activity. As dedifferentiation markers, gamma-glutamyl transpeptidase and alpha-fetoprotein have been studied. From the results, it can be deduced that hepatocytes can be cryopreserved and transplanted and that under these conditions they maintain hepatic features for a long time. Following transplantation, several specific liver functions appear or are enhanced in the spleen. Freshly isolated and cryopreserved transplanted hepatocytes have similar behaviors, although a difference in the expression of the function can be observed.(ABSTRACT TRUNCATED AT 250 WORDS)
Abstract: The histological and ultrastructural study of the ductuli efferentes in epididymides from 40 adult men revealed the occurrence of multinucleate epithelial cells in all specimens. These cells appeared in the luminal protrusions of epithelial folds and correspond to either principal or ciliated cells. The ultrastructure of their cytoplasm did not differ from that of their respective mononucleate cells. Multinucleate cells contained 3-20 closely juxtaposed nuclei, thus appearing much more irregularly outlined than those of the mononucleate epithelial cells. Multinucleation four times more frequent in the principal cells than in the ciliated cells. The number of multinucleate cells increased progressively from the age of 60 years onwards. The average number of nuclei per cell increased in the fourth decade of life, was maintained up to the eighth decade, and then increased again.
Abstract: Whole-mounted preparations of the tunica albuginea of the rat testis were studied using light microscopy techniques for demonstration of cholinergic nerve fibres (Karnovski-Root method), catecholaminergic nerve fibres (De la Torre's method) and actin filaments (avidin-biotin-peroxidase method). An ultrastructural study of different regions of the albuginea was also performed. Cholinergic fibres were seen to penetrate into the albuginea with the testicular artery to form a broad network in the mediastinum testis, many fibres ending beneath the rete testis epithelium. Catecholaminergic fibres penetrated through the middle part of the cauda epididymis and formed a plexus in the albuginea covering the inferior testicular pole. This plexus gave rise to straight fibres which formed varicosities, some of them appeared related with mast cells. Actin-containing cells were only found beneath the rete testis epithelium. These cells were similar to myofibroblasts. The location of both cholinergic fibres and contractile cells among the rete testis channels suggests that these cells may be involved in the pumping of semen towards the ductuli efferentes and that their contractility may be regulated by cholinergic fibres.
Abstract: An immunohistochemical study of laminin, type IV collagen, and fibronectin was carried out in the testes of normal men and in the cryptorchid and contralateral scrotal testes of cryptorchid men from 2 to 40 years of age. The integrated optical density (IOD) per unit area of the lamina propria was measured in the immunostained sections. Fibronectin was found throughout the thickness of the lamina propria of the seminiferous tubules and in the interstitial connective tissue. No differences between normal and cryptorchid testes were found. Laminin was observed in the innermost part of the lamina propria of the seminiferous tubules and surrounding the endothelium of blood capillaries from infancy. No differences were found between normal and cryptorchid testes in the prepubertal period. In adult cryptorchid testes, laminin formed more numerous and deeper invaginations towards the seminiferous epithelium than in normal adult testes. Type IV collagen appeared throughout the thickness of the lamina propria of normal testes as well as in the wall of interstitial blood vessels. From infancy, the lamina propria of seminiferous tubules, but not blood vessel walls, showed lesser immunostaining for type IV collagen and a lower IOD of this component than did control tests from men of the same age. No differences between unilateral and bilateral cryptorchidism were found. The contralateral scrotal testes of cryptorchid males showed intermediate immunostaining for type IV collagen between that of normal control testes and that of cryptorchid testes. These findings suggest that the lamina propria of seminiferous tubules is lesioned at an early age in both cryptorchid and contralateral scrotal testes of cryptorchid men.
Abstract: A quantitative and ultrastructural study was performed on biopsies of human cryptorchid testes to investigate lesions in the lamina propria and Sertoli cells. Prepubertal cryptorchid testes (1-9 years of age) were classified into four groups: Type 1, testes with minimal lesions; Type II, testes with a moderate decrease in tubule diameter and spermatogonal number; Type III, testes with Sertoli cell hypoplasia and a marked reduction in tubule diameter and spermatogonal number; and Type IV, testes with Sertoli cell hyperplasia and a variable reduction in spermatogonal number. An increase in thickness of the lamina propria was found in Type II and III testes from 5 years of age onwards. These testes also showed a decrease in both the average number of peritubular cells per cross-sectioned tubule and in the average nuclear volume of these cells. Most of the postpubertal cryptorchid testes from 13- to 18-year-old youths presented a prepubertal pattern suggestive of delayed testicular maturation. Postpubertal testes from 19- to 27-year-old men were classified into three types: Type A testes showed complete spermatogenesis, mature Sertoli cells and no lesions in the lamina propria; Type B testes showed isolated spermatogonia, mature Sertoli cells, and a marked thickening of the lamina propria; and Type C testes showed isolated spermatogonia, hyperplasia of immature Sertoli cells, and a slightly thickened lamina propria. Maturation of the lamina propria was always associated with maturation of the Sertoli cells. Thickening of the lamina propria was associated with peritubular cell alterations consisting of decreases in the nuclear volume (average and total per testis) of peritubular cells and increases in the number of these cells per cross-sectioned tubule. The three types of adult cryptorchid testes appear to be the postpubertal transformation of Type 1 testes (Type A), Type II and Type III testes (Type B), and Type IV testes (Type C).
Abstract: To determine the mechanics of arteries forming the carotid rete mirabile of goat and cattle, the wall tension-circumference length relationship was recorded isometrically in isolated retial arteries of 350-600 microns in external diameter under relaxed and potassium-activated conditions. The results are compared to those of pial arteries of similar size. In relaxed retial and cerebral vessels, tension-length relationship had nonlinear characteristics and the incremental elastic modulus rose as the circumference length increased, the modulus being lower in retial than in pial arteries. However, the stiffness of the artery considered as a whole (stiffness parameter beta) was similar in the two types of vessels. In activated arteries, force development was a function of length and the magnitude of the response increased until an optimal length was reached after which the response decreased with further stretch. In all stretches studied the developed force was much lower in retial arteries. The calculated active stress at the various lengths was also smaller in retial than in cerebral arteries and the maximum active stress for retial arteries was about 30-50% of that for cerebral arteries. Therefore, the passive mechanical properties of retial arteries closely resemble those of cerebral arteries but retial arteries have a small contractility. This suggests that the carotid rete mirabile of Artiodactyla has a minor role in actively controlling blood supply to the brain. Comments on the hemodynamic significance of the carotid rete are included.
Abstract: The reactivity of the dog cavernous carotid artery to stretch, field electrical stimulation, and norepinephrine was studied using arterial segments under isometric conditions. Light microscopy revealed that this artery is of muscular type and its external surface is covered by venous endothelium, and fluorescence microscopy showed a dense adrenergic innervation. On stretch, arteries exhibited an immediate, transient contraction (phasic response) and a late, maintained contraction (tonic response) that were unaffected by tetrodotoxin (10(-6) M) or endothelium removal but were reduced by the inhibitors of cyclooxygenase indomethacin (10(-6) M), acetylsalicylic acid (3 x 10(-5) M), or meclofenamate (10(-5) M). Electrical stimulation (0.5-4 Hz) contracted the arteries in a frequency-dependent manner, and the response was reduced by tetrodotoxin, phentolamine, (10(-6) M), or the inhibitors of cyclooxygenase used but was unaffected by endothelium removal. Norepinephrine (10(-9)-3 x 10(-4) M) caused dose-dependent contraction that was blocked by phentolamine and by the inhibitors of cyclooxygenase but was not modified by endothelium removal. The results indicate that the dog cavernous carotid artery develops myogenic tone on stretch and contracts on adrenergic stimulation. They also suggest that in these responses prostaglandins but not the endothelium are involved. Therefore, the cavernous carotid artery, because of its location and reactivity, could be of relevance in regulating blood flow or pressure within the cerebral circulation when arterial pressure or adrenergic activity increases.
Abstract: The proliferative activity of human spermatogonia from the fetal period to senility was studied by means of cytophotometric quantification of the nuclear DNA content in histological sections. Proliferating spermatogonia that were replicating or had replicated their DNA (DNA content between 2.5c and 4.5c) were observed in all ages. The percentages of these spermatogonia were high (18.2%) in the second trimester of gestation, decreased in the third trimester (8.2%), maintained similar values in newborns (7.1%) and infants (9.5%), and increased markedly in 4- to 9-year-old children (22.5%). The latter percentage was maintained during puberty (20.1%), decreased significantly in adulthood (17.8%), and was higher in aging testes (25.2%). About 2% of spermatogonia with a DNA content higher than 4.5 c were observed from 4-15 years of age as well as in the testes of elderly men. Sertoli cells replicating their DNA were observed only in fetuses.
Abstract: A histochemical and ultrastructural study of the testes, epididymides and spermatic cords from 6 adult men with amyloidosis secondary to several diseases is described. Amyloid fibrils, consisting mainly of AA protein, were deposited in the walls of blood vessels of the testis, epididymis and spermatic cord, as well as in the tunica propria of the seminiferous tubules. The seminiferous epithelium showed slight hypospermatogenesis. Many Leydig cells exhibited unstained 'bar' or 'arc'-shaped cytoplasmic areas similar to the cholesterol deposits of Leydig cells in adrenoleukodystrophy.
Abstract: The histologic study of testicular specimens from 218 consecutive autopsies revealed a cystic transformation of the rete testis in 26 men. Serial sections of the testes, epididymides, and spermatic cords of these men were studied by light microscopy. In 15 cases, the rete testis dilation was caused by obstruction of the epididymis. In five cases, the dilation was due to obstruction of the intratesticular excretory ducts caused by varicocele. In the remaining six men, no evidence of obstructive processes was found. These six patients had been diagnosed with renal failure and underwent hemodialysis. The rete testis in these men showed dilated channels lined by a high columnar or pseudostratified epithelium. They contained spermatozoa, with a proteinaceous material and calcium oxalate crystals. The ductuli efferents showed eosinophilic granules in their epithelial cells and their lumen contained spermatids, spermatozoa, cell remnants, calcium oxalate crystals, and multinucleate giant cells. These rete testis lesions are similar to those previously reported in the kidney of hemodialyzed patients.
Abstract: We have reported that a liver growth factor isolated from plasma of partially hepatectomized rats is an albumin-bilirubin complex. In this paper, we characterize the liver growth factor purified from subjects with hepatitis (h-LGF). This factor increases synthesis of DNA in a dose-dependent manner both in vivo in mouse hepatocytes, with a dose of maximal stimulation of 150 ng of h-LGF/mouse, and in vitro in rat liver cell culture, with maximal effect at 7.5 to 10 ng of h-LGF/ml. In vivo, h-LGF increases the mitotic index of mouse hepatocytes, its action being organ-specific, acting on liver, but not on spleen, kidney, lung or brain. In vitro, h-LGF stimulates the uptake of 22Na+ by hepatocytes. In addition, we carried out a study comparing it with human serum albumin in terms of absorbance, fluorescence, circular dichroism spectra, amino acid composition, tryptic maps and antigenic determinants (Ouchterlony immunodiffusion). All these tests suggested that human serum albumin is a constituent of h-LGF. Moreover, when albumin isolated from humans without hepatic pathology is incubated with bilirubin, the albumin-bilirubin complex formed mimics the activity of the human liver growth factor with respect to stimulation of DNA synthesis and the effects on the mitotic index of mouse hepatocytes in vivo. We propose that this human liver growth factor is an albumin-bilirubin complex.
Abstract: The occurrence of diverticula in human seminiferous tubules was investigated in the adult human testis in normal as well as physiologic (aging) and pathologic (germ cell depletion, obstruction of male excretory ducts, varicocele and systemic arteriosclerosis) conditions. Diverticula, which are evaginations of the seminiferous epithelium surrounded by a thin tunica propria, were present in all groups studied. The number of diverticula per mm2 testis was higher in the testis with obstruction than in those without obstruction at each age considered. The number of diverticula increased with age in both the obstructed and nonobstructed testis. No changes in the number of diverticula per mm2 testis were found in relation to systemic arteriosclerosis or different degrees of germ cell depletion. Varicocele was only associated with increased numbers of diverticula when it was also associated with obstruction. The formation of diverticula in human seminiferous tubules seems to be an obstructive process related to increasing age.
Abstract: The development of a reproducible, effective system of cryopreservation of hepatocytes would create new possibilities for metabolic studies, as well as the clinical application of hepatocellular transplants in diverse hepatic disorders. For the purpose of dealing with both questions, we have studied different media and procedures of freezing, and have chosen that combination which afforded the best results for long-term studies of function. A series of intrasplenic transplants were performed with freshly isolated hepatocytes as well as cryopreserved ones, and several hepatospecific parameters (Alanine aminotransferase, ornithine carbamoyl transferase, lipogenesis, uptake of 99mTc-N-(p-butylphenylcarbamylmethyl)iminodiacetic acid, G6Pase, albumin, and gamma-glutamyl transpeptidase (gamma-GT) were quantified for 9 months. The histological study of the cells reveals that they acquired an architecture characteristic of the liver. All of the parameters indicative of hepatic function were detected throughout the 9-month period in the transplanted spleens. All the spleens, controls and transplanted, were found negative for the presence of the dedifferentiation marker gamma-GT. These results clearly indicate the possibility of recovering hepatocytes which maintain their specific functions after being subjected to a freezing-thawing process, with the corresponding implications for future clinical application as well as for hepatic biochemistry.
Abstract: Hepatocyte transplantation (HT) is among the most frequently employed approaches in the treatment of liver disease. This work deals with the occurrence of rejection when allogeneic hepatocytes are transplanted into the spleens of rats. Two strains of rats were used: Lewis and F344. The recipients were divided into two groups, one of which received ciclosporin (Cic), while the other received no treatment. After HT, the spleens were removed periodically. The animals that did not receive Cic suffered such intense rejection that 7 days after HT no hepatocytes could be identified in the splenic pulp. On the other hand, hepatocytes could be observed throughout the entire study in the spleens of rats treated with Cic. We conclude that HT is followed by intense rejection, which can be avoided by the administration of Cic.
Abstract: The carotid rete of Artiodactyla, an intracranial arterial plexus which supplies blood to the brain, has intrigued investigators for a long time. This study was designed to examine the responsiveness of isolated retial arteries (250-700 microns in external diameter) of goat, pig, and cattle. The findings in these arteries were compared to those observed in cerebral arteries (250-650 microns in external diameter) of the same animal species. The magnitude of the arterial responses to potassium chloride varied with the resting tension applied to the tissue. The two types of vessels exhibited similar resting tension values (0.3 g) for maximal tension development in response to potassium chloride; however, the ability of retial vessels to contract in the presence of potassium chloride was consistently smaller than that of cerebral arteries. The contractile response of retial arteries to norepinephrine (10(-9) to 10(-4) M), tyramine (10(-8) to 10(-3) M), and field electrical stimulation (2-16 Hz) was negligible. The same retial arteries exhibited dose-dependent contractions in response to 5-hydroxytryptamine (10(-9) to 10(-5) M) and histamine (10(-9) to 3 X 10(-4) M). Cerebral arteries exhibited larger responses to the vasoactive agents than retial arteries. Our findings indicate that retial arteries have a small vasomotor activity in response to adrenergic stimulation or to vasoactive agents. Therefore, the carotid rete of Artiodactyla may have a low ability to change resistance to blood flow under neural or hormonal influences.
Abstract: A new method for correlating testicular biopsies and spermiograms is proposed. The number of spermatogonia, round spermatids, and elongated spermatids per cross-sectioned tubule were calculated in the testes from 33 subfertile men and in 10 control normal testes. According to these quantitations, the testes in subfertile men were classified as testes with maturation arrest of spermatogenesis, testes with hypospermatogenesis, and testes with associated maturation arrest and hypospermatogenesis. A power regression curve correlating the number of elongated spermatids and sperm numbers in the spermiogram was performed.
Abstract: We have reported the purification and characterization of a protein that behaves as a liver growth factor, showing activity either in vivo or in vitro [DÃaz-Gil et al. (1986) Biochem. J. 235, 49-55]. In the present paper, we identify this liver growth factor (LGF) as an albumin-bilirubin complex. This conclusion is supported by the results of chemical and spectroscopic characterization of this protein as well as by experiments in vivo. Incubation of albumin isolated from normal rats with bilirubin/albumin molar ratios (r) resulted (when r = 1 or 2) in a complex with liver DNA synthesis promoter activity identical with that of LGF. The exact amount of bilirubin bound to albumin was assessed by fluorescence and c.d. spectra. This albumin-bilirubin complex showed the same dose-dependence profile as LGF either at low or high dose of protein injected per mouse. Both LGF and albumin-bilirubin complex produced similar increases in the mitotic index of mouse hepatocytes in vivo. A new mechanism for the onset of the hepatic regenerative process is proposed.
Abstract: The appearance of a liver DNA synthesis promoter (HP) in rat plasma after dimethylnitrosamine (DMNA) or thioacetamide injection was investigated. After 48 h, DMNA (30 mg kg-1 body weight) produced liver (centrilobular) necrosis and intense hepatic regeneration, as assessed by microscopic observations of liver slices, as well as augmented transaminase levels; HP was detectable under these conditions. After 5 days, transaminases and HP returned to normal values (the latter undetectable), coinciding with a lack of necrotic zones. At 60 mg DMNA kg-1 body weight, necrotic areas were more marked and transaminases and HP levels higher after 48 h than with the lower dose; these increases were even more pronounced at 90 mg DMNA kg-1 body weight. After thioacetamide injection (200 mg kg-1 body wt) the situation at 48 h was very similar, with focal, centrilobular necrosis, frequent regenerative signs, high transaminases and detectable HP. Rats recovered after 7 days in a similar fashion as with DMNA. At 400 mg thioacetamide kg-1 body weight, necrotic areas and regeneration zones were more widespread and transaminases and HP higher after 48 h than with the lower dose. On account of the differing modes of action of DMNA and thioacetamide in rat liver, it is proposed that the appearance of HP activity in plasma could be related to the regenerative process that follows hepatotoxic damage.
Abstract: The numbers of Ap and Ad spermatogonia per unit section of the testis were calculated in autopsy specimens from young adults and elderly men without testicular pathology. The number of Ap spermatogonia decreased from the 6th decade of life, whereas that of Ad spermatogonia began to decrease in the 8th decade. Although it has been reported that Ad spermatogonia are more sensitive to noxious agents than Ap spermatogonia, the involution of Ap spermatogonia precedes that of Ad spermatogonia. These findings provide new information on concepts relating to the spermatogonia precedes that of Ad spermatogonia. These findings provide new information on concepts relating to the spermatogonial stem cell in man.
Abstract: The numbers of each different cell type in the human seminiferous epithelium were determined throughout the 6 stages of the cycle in both semithin and ultrathin sections obtained from 15 young adult men with normal testicular histology. Up to 4 types of A spermatogonia (Ad, Ap, Al and Ac) were distinguished. In addition, the DNA nuclear content of seminiferous epithelium cells was determined on Feulgen-stained sections. Both Ad and Ap spermatogonia showed a 2c DNA content and were present in the 6 stages of the cycle, though their numbers decreased in stages III-V. Both Al and Ac spermatogonia showed a DNA content varying from 2c to 4c. Al spermatogonia were observed in stages III-V; their numbers plus those of Ad spermatogonia in these stages were similar to the numbers of Ad spermatogonia in the other stages lacking in Al spermatogonia. Ac spermatogonia appeared in stages III-VI and their numbers plus those of Ap spermatogonia in stages III-V were similar to the numbers of Ap spermatogonia in the other stages lacking in Ac spermatogonia. The results suggest that Ad spermatogonia are the stem cells. Some of them replicate their DNA; during this replication they appeared as Al spermatogonia. Al spermatogonia divide, giving rise to both Ad and Ap spermatogonia. Some Ap spermatogonia replicate their DNA; during this process they are transformed into Ac spermatogonia which divide, giving rise to B spermatogonia.
Abstract: Human testicular specimens were obtained from biopsies and autopsies covering the period from birth to adulthood. The number of testosterone-containing Leydig cells was determined using the peroxidase-anti-peroxidase method. This number decreased markedly from 3-6 months of age to the end of the first year of life and, up to 6 years of age, only a small number of testosterone-containing cells was found. From 6 years onwards the number of Leydig cells progressively increased. Ultrastructural examination revealed four types of Leydig cells: fetal-type Leydig cells (from birth to 1 year of age) with round nuclei, abundant smooth endoplasmic reticulum and mitochondria with tubular cristae; infantile-type Leydig cells (from birth to 8-10 years of age), showing a multilobated nucleus, moderately abundant smooth endoplasmic reticulum, some lipid droplets and mitochondria with parallel cristae; prepubertal, partially differentiated Leydig cells (from 6 years of age onwards) with regularly-outlined round nuclei, abundant smooth endoplasmic reticulum, mitochondria with tubular cristae, and some lipid droplets and lipofuscin granules; and mature adult Leydig cells (from 8-10 years of age onwards). The ultrastructure of the infantile-type Leydig cells and the lack of delay between the disappearance of the fetal-type Leydig cells and the appearance of infantile-type Leydig cells suggest that fetal-type Leydig cells give rise to the infantile-type Leydig cells. Before puberty, myofibroblast-like precursor cells differentiate into the prepubertal, partially differentiated Leydig cells, which complete their differentiation into the adult Leydig cells.
Abstract: The testicular connective tissue of patients with alcoholic and non-alcoholic cirrhosis was studied by Picrosirius staining for collagen fibres and electron microscopy, and then compared with the connective tissue of normal testes. Under polarized light, the interstitial connective tissue in the testes of patients with alcoholic and non-alcoholic cirrhosis exhibited a green birefringence which did not appear in normal testes, and the orange birefringence characteristic of the tunica propria in the testes of both cirrhotic and normal males was less developed in cirrhosis. This suggests that a connective tissue, consisting of smaller and less ordered collagen fibre bundles than those in the tunica propria is formed in the testicular interstitium of cirrhotic males, and the collagen fibre bundles of the tunica propria are less developed than in normal testes. The appearance of connective tissue was associated with a decrease in the number of mast cells in the testes of cirrhotic males suggesting the involvement of mast cells in the synthesis, packing, and organization of collagen fibres. The cause of the decrease in mast cell numbers may be related to hormone alterations, in particular testosterone deficiency.
Abstract: A liver DNA synthesis promoter activity was detected in human plasma from subjects with hepatitis. The assay procedure consisted of intraperitoneal injection into mice of aliquots of plasma, previously chromatographed on Sephadex G-25. After 24 hr, [3H]thymidine was injected and its incorporation into liver DNA measured. The increase in [3H]thymidine uptake of injected mice was not detected in those administered plasma from normal subjects (basal [3H]thymidine incorporation was that corresponding to saline-injected mouse values). At a maximal effective dose (0.3 mg protein per mouse), plasma from subjects with hepatitis increased the mitotic index of mouse liver hepatocytes; at the same dose, plasma from normal subjects had no effect. This DNA synthesis promoter activity appears to be a protein, as it is sensitive to trypsin digestion and heat.
Abstract: The number of mononucleate and multinucleate Leydig cells per unit area of the testis was determined in normal adult men using the peroxidase-anti-peroxidase method for testosterone detection. The results of this study indicate that the number of multinucleate Leydig cells increases markedly with age, whereas the total Leydig cell population decreases.
Abstract: This ultrastructural and quantitative study of Leydig cells in Klinefelter's syndrome has been performed using the entire testes obtained from four autopsies plus testicular biopsies from 20 patients. The total Leydig cell volume per testis in Klinefelter's syndrome was similar to that of control testes. However, the total number of Leydig cells per testis was significantly lower in the patients with Klinefelter's syndrome than in the controls. This may be attributed to the Leydig cell hypertrophy that these patients underwent. In patients with Klinefelter's syndrome the average volume of each Leydig cell was significantly higher than that of the controls. Electron microscopic findings revealed that besides ultrastructurally normal Leydig cells there were three other morphologically abnormal Leydig cell types: abnormally differentiated cells; multivacuolated cells; and immature Leydig cells. The ultrastructurally normal Leydig cells were probably the cells principally responsible for testosterone biosynthesis in these patients, whereas the other Leydig cell types were probably barely or not at all functional. The hypertrophy showed by the ultrastructurally normal Leydig cells may be interpreted as an attempt to increase testosterone production.
Abstract: Mast cells are a constant cell-type in the connective tissues of the human testis and epididymis from birth to adulthood. Ultrastructural study shows that these cells are similar to those found in other connective tissues. Histometric studies revealed that the number of mast cells in the interstitium, mediastinum and albuginea of the testis as well as in the epididymal connective tissue increases slightly during infancy, decreases during childhood, and then increases again at puberty. Increases at puberty are particularly evident in both the testicular interstitium and the epididymis. During adulthood, the number of mast cells progressively decreases in all testicular and epididymal connective tissues. Changes in mast cell number may be related to changes observed in the development of testicular connective tissue which occurs primarily during infancy and puberty.
Abstract: Hormone measurements, spermiograms and testicular biopsies studies were performed in young with varicocele. In addition, the testes and epididymides of 27 adults with varicocele were obtained from autopsies. Light and electron microscopic examination of biopsy and autopsy specimens revealed two types of lesions in testes with varicocele: 1) a diffuse lesion consisting of abnormal spermatozoa and spermatid morphology and sloughing of immature spermatozoa and spermatid; 2) focal lesion, distributed irregularly throughout the testicular parenchyma, affecting several small groups of seminiferous tubules. Each of these groups corresponded to a testicular lobule and showed different degrees of tubular atrophy, so that the focal lesions were distributed in a mosaic pattern. The testicular interstitium showed dilated veins and venules, and progressive collagenization. Some testes showed dilated veins in the rete testis, which compressed several tubuli recti and caused tubular atrophy in the seminiferous tubules opening into these tubuli recti. Other testes showed dilated young veins among the ductuli efferentes, and the rete testis channels appeared to be dilated. Among the different etiological mechanisms which have been suggested to for testicular lesions in varicocele, tubular obstruction at the level of either the tubuli recti or the ductuli efferentes might be responsible for lesions leading to testicular atrophy.
Abstract: The distribution of elastic fibres in the human ductus deferens from birth to senility was studied by light and electron microscopy. Elastic fibres are lacking in the ductus deferens in infants and children. In the adult ductus deferens, they form two layers in the lamina propria: (1) an inner layer of circumferentially oriented elastic fibres, and (2) an outer meshwork of elastic fibres. Elastic fibres are also present in the narrow intercellular spaces between the smooth muscle cells of the muscular coat, mainly in the inner muscular layer. A layer of elastic fibres surrounds the muscular coat. The ductus deferens of ageing subjects shows fragmentation and disorganisation of the elastic fibre layers of the lamina propria. Elastic fibres in the muscular coat are more abundant than in younger adults, forming larger bundles. Electron microscopy demonstrated the presence of immature elastic fibres at puberty as collections of microfibrils, some of them containing loci of amorphous substance (elastin). In the adult ductus most elastic fibres have a mature appearance. The amount of amorphous substance has increased and the number of microfibrils has decreased. Electron-dense inclusions are present within the amorphous substance. With advancing age the amorphous substance forms large, structureless masses showing abundant electron-dense inclusions and areas of rarefaction. A thin layer of microfibrils is present only at the periphery of the elastic fibres. Whether or not androgenic hormones are in any way involved in the formation of elastic fibres in the ductus deferens and testis is something which requires further study.
Abstract: Testicular biopsy specimens from adult patients affected with cryptorchidism, Klinefelter's syndrome, and Del Castillo's syndrome were examined by light and electron microscopy. The study revealed a high proportion of testes showing seminiferous tubules with hyperplasia of Sertoli cells (from 25 to 45 cells per transverse tubular section). These cells had an immature appearance and showed a pseudostratified distribution. The nucleus was round to ovoid and regular in outline, with a smaller nucleolus than that of mature Sertoli cells. The cytoplasm showed less development of the endoplasmic reticulum as well as of the secondary lysosomes and lipid droplets than that in mature Sertoli cells. Characteristic features of these immature Sertoli cells were abundant cytoplasmic microfilaments, elaborate interdigitations between adjacent cells, and extensive tight junctions, from basement membrane to lumen. In the cryptorchid testes, a more immature Sertoli cell was found to constitute the majority of the cells in hypoplastic zones. In Klinefelter's and Del Castillo's syndromes as well as in cryptorchid testes to a lesser degree, a transitional type of cell-from immature to mature-was also observed. These observations suggest that Sertoli cells in these primary testicular disorders reflect a congenital deficiency producing abnormal development.
Abstract: The most relevant issues about the histophysiology of human prostate and seminal vesicles are developed in the present chapter.
Respecting to prostate, there was studied important events related to embryology and development, prostate regionalization and its implications on benign and malignant prostate changes; the important role played by innervation and neuroendocrine system on both normal and pathologic prostate; and the distribution and quantification of prostate microvessels and their interest in normal and pathological angiogenesis.
In relation to seminal vesicles, a timely revision of histology, immunohistochemistry, and function of seminal vesicles and their role on male fertility it has been developed.
Abstract: Cadmium chloride (d) is a toxicant that has been implicated in human prostate cancer (PCA). The goal of the present study was to evaluate the immunoexpression of markers for cell proliferation, apoptosis, resistance to apoptosis, and to determine mutations on segments of the bcl-2 gene, in preneoplastic lesions induced in rat prostate after treatment with Cd alone or in combination with zinc chloride (Zn). We evaluated: I) The % of cells positively immunostained for the proliferating cell nuclear antigen (PCNA), 2) The % of apoptotic cells (evaluated by TUNEL), 3) The volume fraction of Bcl-2 immunostaining. 4) The mutations on a segment of 253 pb of bcl-2, in the ventral prostate lobe of normal and treated rats with Cd alone or in the presence of Zn in the drinking water for 18 mo. Our results indicate that the % of PCNA positive nuclei was significantly increased in preneoplastic prostatic acini of Cd-treated rats alone and in combination with Zn, compared to the normal acini of untreated animals.
No significant changes were detected on the apoptotic rate or the volume fraction of Bcl-2. Moreover, no significant changes in the band pattem of the amplified segment of bcl-2 gene were observed after Cd treatment. In summary, our data indicate that, prostate dysplasia induced in rats by Cd increases proliferative activity, without significant changes in either
apoptosis or bcl-2 immunoreactivity.
Abstract: Prostatic intraepithelial neoplasia (PIN) is considered a precursor of prostate cancer (PCA). In the majority of the PINs, the atypia is more marked in basal than lurninal nuclei. The aim of this study was to quantitate the differences in the stereological estimation of mean nuclear volume weighted by volume (Vv nuc) between basal and luminal cells of PIN, in relation to normal prostate epitheliurn, and PCA. Ten PIN cases were compared with 50 PCA, and 5 normal prostates. AIl the specimens were routinely processed and stained with haematoxylin-eosin. The mean±SD of Vv nuc was evaluated for all the groups. The epitheliurn of both normal and PIN specimens was segmented in basal and lurninal compartments, and the Vv nuc measured in both strata. Comparisons among groups were performed by ANOVA (p < 0.05). The value of Vv nuc was significantly lower in normal epithelium compared to both PIN and PCA. Basal layer PIN Vv nuc was significantly higher in comparison to the luminal stratum. Luminal PIN Vv nuc was similar to Vv nuc of PCA: The similarities in nuclear size between PIN and PCA are according to the pre-malignant character commonly assigned to PIN .The increase of basal layer PIN Vv nuc may indicate that the changes heralding the progression from PIN to PCA are produced in this layer, whereas the nuclear features of luminal layer are the same to those of the carcinoma.
Abstract: The present study leads to the conclusion that there are glucagonergic receptors in the liver which may have a haemodynamic role in addition to the metabolic one. There are also glucagon receptors in the smooth muscle of the jejunum and renal vascular bed whose distribution confirms a direct role for glucagon in the motility and tone of the gastrointestinal and vascular smooth muscle fibre as well as its vasoactive role.
There is a heterogenicity between the splanchnic smooth muscle (vessels and walls of hollow organs) and the smooth muscle of the peripheral vessels (femoral bed) in the sense that at this latter location the role played by glucagon must be a limited one, judging from the scarce number of specific receptors. The presence of glucagonergic receptors in the intramural neurons of the
intestine leads us to suspect the possible existence of a preganglionic-blocking effect of glucagon which would inhibit intestinal motility. However, up to date it has been impossible to demonstrate the existence of glucagonergic neurons or nervous fibres by immunohistochemical techniques, so it is quite likely that the neuron receptors for glucagon may not be related to synapses where this hormone would act as a neurotransmitter.
Abstract: Blood supply to the brain of many mammals, notably Cetacea, Sirenia, and Artiodactyla, occurs via an arterial plexus, the rete mirabile. In Artiodactyla the carotid rete lies within the cranium in the cavemous sinus and is interposed in series between the carotid vessels and the circle of Willis.
The presence of nerve fibers associated to the rete has led to the hypothesis that this vascular formation could play an active role in the regulation of cerebral circulation by means of vasomotor reflexes. However, there is no conclusive evidence that retial arteries are well innervated. In the present work, retial arteries of goat, pig, and cattle (three species of the order Artiodactyla) are examined to determine their degree of innervation using transmission electron and fluorescence microscopy. In addition, we present a histomorphometric analysis of retial vessels using light microscopy. The results are compared to those from cerebral arteries of similar size.
Abstract: In Artiodactyla the carotid rete (rete mirabile caroticum) is a plexus of medium-sized arteries which lies within the cavemous sinus and is interposed in series between the carotid and brain vessels. It has been suggested that the rete plays an active role in the regulation of cerebral circulation by means of vasomotor reflexes. However, there is no conclusive evidence of this proposal.
In the present work, isolated arteries from the carotid rete of three species (goat, pig and cattle) of the order Artiodactyla were examined to determine their ability to constrict or dilate in response to field electrical stimulation. In addition, responses of isolated retial arteries were compared to those from cerebral arteries of similar size.
