Profile of NF-κBp(65/NFκBp50) among prostate specific antigen sera levels in prostatic pathologies.

AIM: The aim of this work was to characterise the immunoexpression of NF-κB (p50/p65) in human prostatic pathologies and to study its profiles of activation among sera prostate specific antigen antigen (PSA) according the three groups: 0-4ng/mL, 4-20ng/mL and >20ng/mL. PATIENTS AND METHODS: Twenty-four men with benign prostate hyperplasia (BPH); 19 men with prostate cancer (PC) and five men with normal prostates (NP). Immunohistochemical and western blot analysis was performed. Serum levels of PSA were assayed by immulite autoanalyser. RESULTS: In BPH and PC samples, immunoexpressions were observed for NF-κBp65 and NF-κBp50; while in NP samples, only were detected NF-κBp50. PC samples showed immunoreactions to NF-κBp65 and NF-κBp50 more intense (respectively 24.18±0.67 and 28.23±2.01) than that observed in BPH samples (respectively18.46±2.04 and 18.66±1.59) with special localisation in the nucleus. Different profiles of NF-κBp65 immunoexpressions were observed and BPH patients with sera PSA levels between 0-4ng/mL presented a significant weak percentage compared to BPH patients with sera PSA levels between 4-20ng/mL and >20ng/mL. No immunoreactions to NF-κBp65 were observed in PC patients with sera PSA levels between 4-20ng/mL. CONCLUSION: The sensibility of both NF-κB and PSA to inflammation allowed confirming the relationship between these two molecules and its involvement in prostatic diseases progression (inflammatory and neoplasic).

Innominate truncal dissection and rupture into right pleural cavity following acute type A dissection of the aorta with right coronary ostial avulsion and inferior STEMI.

An innominate truncal dissection and rupture into the right pleural cavity with massive hemothorax is the initial presentation in this 66-year-old lady with type A dissection of the aorta complicated by right coronary ostial avulsion and inferior STEMI. She underwent supracoronary interposition graft replacement of the ascending aorta and hemiarch, interposition graft replacement of the innominate trunk and saphenous vein bypass grafting of the right coronary artery successfully. Innominate truncal rupture following aortic dissection is practically unknown and has not been described before in the absence of aortic rupture. Innominate truncal rupture secondary to other pathologies presents with supraaortic and mediastinal hematomas, but almost never with right hemothorax. On the backdrop of this unusual presentation with no neurological injury, we review the literature for innominate truncal dissection and rupture, other etiologies for innominate truncal rupture, the complex interplay of factors determining neurological injury and discuss the changes in the strategies and conduct of arterial return during cardiopulmonary bypass and selective antegrade perfusion imposed by this previously undescribed instance of innominate truncal rupture due to dissection.

Cypermethrin effects on the adult mice seminal glands.

The insecticide cypermethrin acts upon the sodium channels. Their effects over animal health are not understood. Here, the effects of cypermethrin on the seminal glands (SGs) are studied (1/5 DL50 i.p.). Forty-five adult mice (CF1) were distributed in three groups: (1) untreated, (2) vehicle (oil), and (3) experimental (cypermethrin in oil). The animals were sacrificed at 1 and each 8.6 days. The SGs were processed for histology: Haematoxylin/P.A.S, Thyonin (0.6%) and Immunohistochemistry (Ki-67). In the SGs was quantified: the epithelium height, mastocytes, and cell proliferation. In the results, cypermethrin exerts an intense effect on epithelium height and cell proliferation. A net increase of both parameters was observed at 24 h (p0.05). However, the mastocytes increased drastically and progressively during the experimental period (p0.05). Then, the effects have acute manifestations, which would be responsible for the potential changes in the male's reproductive potentiality.

α-PD-1 therapy elevates Treg/Th balance and increases tumor cell pSmad3 that are both targeted by α-TGFß antibody to promote durable rejection and immunity in squamous cell carcinomas.

BACKGROUND: Checkpoint blockade immunotherapy has improved metastatic cancer patient survival, but response rates remain low. There is an unmet need to identify mechanisms and tools to circumvent resistance. In human patients, responses to checkpoint blockade therapy correlate with tumor mutation load, and intrinsic resistance associates with pre-treatment signatures of epithelial mesenchymal transition (EMT), immunosuppression, macrophage chemotaxis and TGFß signaling. METHODS: To facilitate studies on mechanisms of squamous cell carcinoma (SCC) evasion of checkpoint blockade immunotherapy, we sought to develop a novel panel of murine syngeneic SCC lines reflecting the heterogeneity of human cancer and its responses to immunotherapy. We characterized six Kras-driven cutaneous SCC lines with a range of mutation loads. Following implantation into syngeneic FVB mice, we examined multiple tumor responses to α-PD-1, α-TGFß or combinatorial therapy, including tumor growth rate and regression, tumor immune cell composition, acquired tumor immunity, and the role of cytotoxic T cells and Tregs in immunotherapy responses. RESULTS: We show that α-PD-1 therapy is ineffective in establishing complete regression (CR) of tumors in all six SCC lines, but causes partial tumor growth inhibition of two lines with the highest mutations loads, CCK168 and CCK169. α-TGFß monotherapy results in 20% CR and 10% CR of established CCK168 and CCK169 tumors respectively, together with acquisition of long-term anti-tumor immunity. α-PD-1 synergizes with α-TGFß, increasing CR rates to 60% (CCK168) and 20% (CCK169). α-PD-1 therapy enhances CD4 + Treg/CD4 + Th ratios and increases tumor cell pSmad3 expression in CCK168 SCCs, whereas α-TGFß antibody administration attenuates these effects. We show that α-TGFß acts in part through suppressing immunosuppressive Tregs induced by α-PD-1, that limit the anti-tumor activity of α-PD-1 monotherapy. Additionally, in vitro and in vivo, α-TGFß acts directly on the tumor cell to attenuate EMT, to activate a program of gene expression that stimulates immuno-surveillance, including up regulation of genes encoding the tumor cell antigen presentation machinery. CONCLUSIONS: We show that α-PD-1 not only initiates a tumor rejection program, but can induce a competing TGFß-driven immuno-suppressive program. We identify new opportunities for α-PD-1/α-TGFß combinatorial treatment of SCCs especially those with a high mutation load, high CD4+ T cell content and pSmad3 signaling. Our data form the basis for clinical trial of α-TGFß/α-PD-1 combination therapy (NCT02947165).

TNF/IL-1/NIK/NF-kappa B transduction pathway: a comparative study in normal and pathological human prostate (benign hyperplasia and carcinoma).

AIMS: Tumour necrosis factor (TNF)-alpha induces death or cell proliferation by activation of nuclear factor (NF)-kappaB, also activated by interleukin (IL)-1 alpha. The aim was to investigate upstream and downstream components of NIK transduction pathway in normal (NP), benign prostatic hyperplasia (BPH), prostatic intraepithelial neoplasia (PIN) and prostatic carcinoma (PC). METHODS AND RESULTS: Immunohistochemistry and Western blotting were performed. In NP, the cytoplasm of epithelial cells was intensely immunoreactive to IL-1 receptor-associated kinase (IRAK), TNF receptor-associated factor (TRAF)-6, NF-kappaB inducing kinase (NIK), I kappa kappa alpha/beta, I kappaB alpha and p-I kappaB; weakly to NF-kappaB-p50; and negative to NF-kappaB-p65. BPH samples were intensely immunoreactive to IRAK, TRAF-6, NIK, I kappa kappa alpha/beta, I kappaB alpha, p-I kappaB; weakly to NF-kappaB-p50 and NF-kappaB-p65. Whereas low-grade PIN showed intermediate results between NP and BPH, results in high-grade PIN were similar to those found in PC (low Gleason). In PC, immunoreactivity was intense for IRAK, TRAF-6, NIK, I kappa kappa alpha/beta (increasing with Gleason), I kappaB alpha, p-I kappaB (decreasing with Gleason); weak for NF-kappaB-p50 and NF-kappaB-p65 (decreasing with Gleason). Nuclear NF-kappaB was observed in PC. CONCLUSIONS: NF-kappaB enhances cell proliferation, but also ATF-2 or Elk-1. Since IL-1 and TNF-alpha are related to inflammation and their immunoexpression increases in PC, inhibition of these cytokines might be a possible target for PC treatment, because they decrease the activity of all transduction pathway members that activate transcription factors such as NF-kappaB, Elk-1 or ATF-2.

Clinical outcomes and peritoneal histology in patients starting peritoneal dialysis are related to diabetic status and serum albumin levels.

Peritoneal morphological changes seem to be related to dialysis solutions bioincompatibility and to infections, but the uremic milieu per se may also contribute to peritoneal changes. The influence of diabetes and diabetes-associated comorbidities on peritoneal histological changes in the pre-dialysis stage have been insufficiently studied. The aim of this study is to analyze the effect of diabetes and serum albumin levels on peritoneal histology and certain clinical variables such as peritoneal permeability, technique failure, and general mortality in patients starting peritoneal dialysis (PD) treatment. Eighteen PD patients without diabetes (uremic non-diabetic group, U-ND) and 65 with diabetes (uremic diabetic group, U-D) were studied prospectively. Clinical and biochemical variables were registered, and a parietal peritoneum biopsy was obtained at the time of the peritoneal catheter placement. Peritoneal histology was evaluated by light microscopy and immunohistochemistry. A control group of 15 non-uremic, non-diabetic (NU-ND) patients who underwent non-complicated elective abdominal surgery was also studied and used as control. The proportion of patients with peritoneal morphological changes as evaluated by light microscopy was higher in the two groups of uremic patients than in the control. The U-D group had higher mesothelial loss (40.9 vs 29.4%), higher mesothelial basement membrane thickening (45.5 vs 23.5%), higher proportion of vascular wall thickening/sclerosis (39.7 vs 11.1%), and higher proportion of inflammatory infiltrate (45.4 vs 23.6%) than the U-ND group. Uremic patients had lower density of mesothelial cells and higher density of inflammatory cells than the control, as evaluated by immunohistochemistry. These changes were even more striking in the U-D group than in the U-ND group. On the other hand, inflammatory infiltration to the peritoneum, mesothelial cell loss, and mesothelial basement membrane thickening were associated with higher technique failure and mortality. However, when the serum albumin level was introduced into the model, the aforementioned associations became nonsignificant. In conclusion, uremia and diabetes were associated with important peritoneal histological changes before starting PD treatment. Diabetes associated with uremia was more strongly related to the peritoneal changes than uremia per se. Hypoalbuminemia and peritoneal inflammatory infiltrate were markedly associated with technique failure and mortality in patients starting PD treatment.

Echocardiographic, electrocardiographic and blood pressure changes induced by icodextrin solution in diabetic patients on peritoneal dialysis.

The use of icodextrin as an osmotic agent in solutions for peritoneal dialysis (PD) has important cardiovascular effects related with better control of extracellular volume. Among them, reduction of arterial pressure and an improvement in echocardiographic parameters stand out. In diabetic patients, icodextrin has additional potential advantages related with better metabolic control. In a multicenter, open-label randomized controlled trial, the effects of icodextrin solutions were compared to glucose solutions on echocardiographic, electrocardiographic, and blood pressure changes in diabetic patients on PD. Two phases were noted in the follow-up. In the early phase (6 months), reduction in ambulatory blood pressure (ABP) and left ventricular end diastolic diameter were found in the icodextrin group. These changes correlated with changes in body fluids. In the late phase (12 months), a trend towards baseline values in ABP was seen. Changes in inferior vena cava diameter and in low frequency R-R variability spectral analysis in the icodextrin group suggest that icodextrin increases circulating blood volume and sympathetic tone, probably by accumulation of icodextrin metabolites in the bloodstream and improvement in diabetic neuropathy as a result of lower peritoneal glucose absorption. The effects of icodextrin in diabetic patients were related to better fluid management and metabolic control.

Nebulin-like protein in the earthworm Eisenia foetida. Immunocytochemical electron microscopic study and western blot analysis of several muscle cell types.

Nebulin is a giant protein (500-900 kDa), which has been reported only in the skeletal muscle (not in cardiac muscle) of vertebrates. The possible presence and distribution of nebulin-like proteins in obliquely striated muscles (body wall and inner muscular layer of the pseudoheart) and smooth muscle (outer muscular layer of the pseudoheart) from the earthworm Eisenia foetida have been examined by means of Western blotting analysis and immunoelectron microscopy, using antibodies against mouse nebulin. The results were compared with those obtained in skeletal, cardiac and smooth muscles of the mouse. In the mouse, immunoreaction to nebulin was observed only in the skeletal muscle and extended along the length of the thin filament. In the earthworm, immunoreaction to a nebulin-like protein was found in the muscle of the body wall and the inner muscular layer of the pseudoheart, but not in the outer muscular layer of the pseudoheart. By electron microscopy, immunolabeling to this protein was observed along the whole length of the thin filament. Western blotting analysis of this nebulin-like protein showed a single band at an estimated molecular mass between 350 and 450 kDa that is slightly lower than that of mouse skeletal muscle nebulin.

Immunocytochemical electron microscopic study and Western blot analysis of caldesmon and calponin in striated muscle of the fruit fly Drosophila melanogaster and in several muscle cell types of the earthworm Eisenia foetida.

Caldesmon and calponin are two proteins that are characteristic of vertebrate smooth muscle. In invertebrates, caldesmon has only been studied in some molluscan muscles, and no previous references to calponin have been found. The aim of this paper was to investigate the presence and distribution of caldesmon and calponin in several invertebrate muscle cell types, classified according to their ultrastructural pattern: transversely striated muscle (flight muscle from Drosophila melanogaster), obliquely striated muscle (muscular body wall and inner muscular layer of the pseudoheart from the earthworm Eisenia foetida), and a muscle of doubtful classification which seems to be intermediate between smooth muscle and obliquely striated muscle (outer muscular layer of the pseudoheart, from E. foetida), using electron microscopy immunocytochemistry and Western blot analysis. Immunoreactions to both caldesmon and calponin were observed in the outer muscular layer cells from the earthworm pseudoheart but neither in the transversely striated muscle of D. melanogaster nor in the obliquely striated muscle from the earthworm. Present findings suggest that caldesmon- and calponin-like proteins are also present in invertebrate muscle cells, but only in those that are ultrastructurally similar to the vertebrate smooth muscle cells. Since discrepancies in the classification of some invertebrate muscles are common in the literature, the use of distinctive markers, such as troponin, caldesmon and calponin may improve our understanding of the nature and properties of many invertebrate muscles showing an ultrastructural pattern that does not resemble any of the classic muscle types.

Leukemia inhibitory factor and ciliary neurotropic factor promote the survival of Sertoli cells and gonocytes in coculture system.

Leukemia inhibitory factor (LIF) and ciliary neurotropic factor (CNTF) were found to be pleiotropic modulators of Sertoli cell and gonocyte development (both isolated from the neonatal rat testis) in a coculture system, whereas IL-6, another member of this cytokine family, had no effect on these cells. LIF and CNTF significantly enhanced the survival of the Sertoli cells in a dose- and time-dependent manner. The effect of LIF on the Sertoli cells was significant at a concentration of 1 ng/ml after 3 or 6 days of culture, whereas CNTF had a significant effect at 10 ng/ml. Neither LIF nor CNTF had an effect on Sertoli cell proliferation. The survival of proliferating gonocytes (isolated from 3-day-old rats testes) was also significantly higher in cultures to which LIF (7.5 ng/ml) or CNTF (10 ng/ml) was added. No effect of these cytokines was found on the mitotic activity of proliferating gonocytes. However, LIF (7.5 ng/ml) stimulated the proliferation of quiescent gonocytes (isolated from day 1 testes) after 3 days of culture. Combinations of LIF (or CNTF) with fibroblast growth factor 2 (10 ng/ml) and steel factor (50 ng/ml) did not further improve the long term culture of the gonocytes. LIf- and CNTF-like proteins of the expected molecular masses (32,000 and 22,000 daltons, respectively, under reducing conditions) were found by Western blotting in testicular extracts of 3-day-old rats. Taken together, these results indicate that LIF or CNTF may play a role at the start of the spermatogenesis. The characterization of receptors for LIF or CNTF on the gonocytes and/or neonatal Sertoli cells will aid in a better understanding of the physiological role of these cytokines in the reproductive system.

Oncostatin M in the normal human testis and several testicular disorders.

The immunohistochemical reaction to oncostatin M (OSM) was studied in normal human testes at different ages (fetuses, newborns, children, pubertal boys, adults, and elderly men), as well as in several testicular disorders including carcinoma-in-situ cells (CIS), germ cell tumors, benign functioning Leydig cell tumor, androgen insensitivity syndrome, Klinefelter's syndrome, and cryptorchidism. Positive OSM immunostained Sertoli cells were only observed in fetuses. In normal testes, intense OSM immunoreaction was found in the Leydig cells of fetuses, newborns, and adults. Leydig cell immunoreaction was weak in elderly men and absent in children and pubertal boys. In some testicular disorders (Leydig cell tumor, cryptorchidism, and CIS), Leydig cell immunoreaction was as intense as in normal adult testes. This immunoreaction was heterogeneous in androgen insensitivity syndrome and was absent in Klinefelter's syndrome and intratubular seminoma. No recognizable Leydig cells were observed in the other testicular tumors. The findings of our study suggest that, in humans, the down-regulation of OSM immunoexpression in Sertoli cells occurs early, and that OSM immunoreaction in the Leydig cells is associated with functionally active and differentiated Leydig cells.

Expression of ghrelin and its functional receptor, the type 1a growth hormone secretagogue receptor, in normal human testis and testicular tumors.

Ghrelin, the endogenous ligand for the GH secretagogue receptor (GHS-R), has been primarily linked to the central neuroendocrine regulation of GH secretion and food intake, although additional peripheral actions of ghrelin have also been reported. In this context, the expression of ghrelin and its cognate receptor has been recently demonstrated in rat testis, suggesting a role for this molecule in the direct control of male gonadal function. However, whether this signaling system is present in human testis remains largely unexplored. In this study we report the expression and cellular location of ghrelin and its functional receptor, the type 1a GHS-R, in adult human testis. In addition, evaluation of ghrelin and GHS-R1a immunoreactivity in testicular tumors and dysgenetic tissue is presented. The expression of the mRNAs encoding ghrelin and GHS-R1a was demonstrated in human testis specimens by RT-PCR, followed by direct sequencing. In normal testis, ghrelin immunostaining was demonstrated in interstitial Leydig cells and, at lower intensity, in Sertoli cells within the seminiferous tubules. In contrast, ghrelin was not detected in germ cells at any stage of spermatogenesis. The cognate ghrelin receptor showed a wider pattern of cellular distribution, with detectable GHS-R1a protein in germ cells, mainly in pachytene spermatocytes, as well as in somatic Sertoli and Leydig cells. Ghrelin immunoreactivity was absent in poorly differentiated Leydig cell tumor, which retained the expression of GHS-R1a peptide. In contrast, highly differentiated Leydig cell tumors expressed both the ligand and the receptor. The expression of ghrelin and GHS-R1a was also detected in dysgenetic Sertoli cell-only seminiferous tubules, whereas germ cell tumors (seminoma and embryonal carcinoma) were negative for ghrelin and were weakly positive for GHS-R1a. In conclusion, our results demonstrate that ghrelin and the type 1a GHS-R are expressed in adult human testis and testicular tumors. Overall, the expression of ghrelin and its functional receptor in human and rat testis, with roughly similar patterns of cellular distribution, is highly suggestive of a conserved role for this newly discovered molecule in the regulation of mammalian testicular function.

Granulomatous epididymal lesion of possible ischemic origin.

In a histologic review of adult epididymides obtained at autopsy (both epididymides of 408 men) or during surgery (261 men with testicular or epididymal nontumoral pathology), a peculiar granulomatous lesion was observed in two autopsy specimens (unilateral) and three surgical specimens. The lesion was located in the caput epididymidis and consisted of a zone of necrosis that involved efferent ducts and interstitial connective tissue and was not associated with an acute inflammatory response. Immunohistochemical study with anticytokeratin antibodies showed the presence of some epithelial cells in the damaged efferent ducts. At the periphery of the lesion, where damage was less severe, the efferent ducts only showed partial necrosis of their wall through which the necrotic material was released to the ductal lumen. Inflammatory infiltrates were scanty and consisted of lymphocytes and CD68-positive macrophages. Lymphocytes were mainly located around the necrotic zone or surrounding the adjacent, well-preserved efferent ducts, whereas macrophages formed large clusters in the ductal lumen. In these clusters, cholesterol crystals and giant cells of foreign body type were frequent. Intratubular epithelial regeneration as well as proliferation of small ducts showing epithelial regeneration and numerous spermatozoa in their lumen were observed. Ceroid granulomata, spermatic granulomata, and epidermoid metaplasia of the efferent ducts were observed in some cases. On the basis of the histologic study, the following developmental stages of the lesion are suggested: ischemic necrosis, granulomatous reaction, cicatrization, and sequelae. The term "granulomatous ischemic lesion" is proposed to designate this reactive lesion.

Cystic transformation of the rete testis.

In a review of the testicular and epididymal specimens obtained from autopsies (1,798 men) or surgery (518 men), cystic transformation of the rete testis (CTRT) was found in 20 autopsies and 18 surgical specimens. When both testes were studied (autopsies), the lesion was bilateral. Ultrasonography revealed a widened mediastinum testis showing small hypoechoic areas. Arteriography showed thin or irregularly outlined testicular arteries, and the epididymal artery was lacking or appeared stenosed. Simple CTRT (without epithelial alteration) was found in both testes of 17 autopsied patients (all were elderly men) and in eight surgically removed testes from patients with sarcoma, tuberculous orchidoepididymitis, or hematocele. The most frequent epididymal lesion was bilateral efferent duct atrophy. In three patients, the rete testis presented nodular proliferation of calcifying connective tissue. CTRT with columnar transformation of the rete testis epithelium was observed in both testes from three patients with alcoholic cirrhosis, and in 10 surgically removed testes from patients with testicular tumor, cryptorchidism, or nonspecific orchitis. In cirrhotic patients, the efferent ducts appeared atrophied. In patients with testicular tumors, the efferent ducts were infiltrated by carcinoma in situ cells (CISs) and often contained granular material, cell debris, or hyaline globules. In both kinds of CTRT (without or with epithelial metaplasia), the most frequent seminiferous tubule lesions were tubular ectasia, hypospermatogenesis, tubular sclerosis, spermatogonium arrest, and sloughing of immature germ cells (spermatids and spermatocytes). The mechanism leading to CTRT might be mechanic (compression of the epididymis by an epididymal tumor or a spermatic cord tumor, or the result of a long-standing epididymitis or traumatic hemocele); ischemic (autopsied elderly men); hormonal (cirrhotic patients); malformative (cryptorchidism); or unknown (the remaining cases).

Epithelial-myoepithelial tumor of the bronchus.

A primary bronchial tumor with a histological pattern similar to that of epithelial-myoepithelial tumor of the salivary gland is reported in a 55-year-old woman. The tumor was well delimited, although not encapsulated, and showed a polypoid growth. The tumor was composed of two types of neoplastic cells: epithelial cells displaying tubules and myoepithelial cells that either formed compact masses or surrounded the tubular formations. Immunohistochemical study confirmed positive immunoreaction to both high- and low-molecular-weight cytokeratins in the epithelial cells and positive immunoreaction to vimentin, S-100 protein, and myosin in the myoepithelial cells.

Testicular biopsy in patients with obstructive azoospermia.

The present report studies the testicular biopsy lesions (histologic and semiquantitative) in a series of 48 patients with obstructive azoospermia of known etiology (vasectomy, congenital absence of vas deferens, herniorrhaphy, hydrocelectomy, Young's syndrome, and ejaculatory duct obstruction) in order to establish objective testicular data that permit the pathologist to diagnose an obstructive process, which should not be mistaken with a primary testicular lesion. The semiquantitative study included determinations of the average numbers of spermatogonia, primary spermatocytes, young spermatids (Sa + Sb), and differentiated spermatids (Sc + Sd). According to this study, the testes were classified into the following groups: (1) normal testes whose germ cell numbers were within normal limits (27 testes); (2) testes with lesions in the adluminal compartment; these lesions comprise two subgroups: (2a) late sloughing of primary spermatocytes (both spermatid types were greatly reduced in number while the other germ cell types were in normal numbers) (45 testes); and (2b) early sloughing of primary spermatocytes (normal spermatogonial number, reduced number of spermatocytes, and scanty spermatids) (9 testes); and (3) lesions in the basal compartment; these lesions comprise two subgroups: (3a) pure hypospermatogenesis (a proportionate decrease in the numbers of all germ cell types) (8 testes); and (3b) hypospermatogenesis associated with sloughing of primary spermatocytes (decreased numbers of all germ cell types with a very scanty number spermatids) (4 testes). Two testes appeared hyalinized and one testis was removed owing to cryptorchidism. The most frequent testicular lesion observed (alteration in the adluminal compartment of seminiferous tubules) seems to be related to the increase in hydrostatic pressure in the tight compartment formed by seminiferous tubules, rete testis, efferent ducts, the epididymal duct, and the initial portion of the vas deferens. The severity of the lesions is probably related to the cause and span of the obstruction. In addition, two azoospermic men without obstructive azoospermia and whose testicular biopsy study revealed meiotic anomalies (with the subsequent bad prognosis) were also studied for comparison. The semiquantitative study of these patients permitted the differential diagnosis between two lesion types. Testes with meiotic anomalies had a disproportionately elevated number of primary spermatocytes, and an extremely low number of young spermatids.

Identification of N- and O-linked oligosaccharides in the human epididymis.

The oligosaccharide sequences of glycoconjugates in the human normal epididymis and the nature of linkages were studied with lectin histochemistry. The usual terminal sequences of oligosaccharide side chains in epithelial cell secretions were Neu5Ac2,3Galbeta1,3GalNAc; SO4Galbeta1,3GalNAc; and Galbeta1,4GlcNAc, and they were mainly found in O-linked glycoproteins. The lectin pattern of mitochondria-rich cells differed from that of principal cells.

Variations in dystrophin complex in red and white caudal muscles from Torpedo marmorata.

We present an up-to-date study on the nature, at the protein level, of various members of the dystrophin complex at the muscle cell membrane by comparing red and white caudal muscles from Torpedo marmorata. Our investigations involved immunodetection approaches and Western blotting analysis. We determined the presence or absence of different molecules belonging to the dystrophin family complex by analyzing their localization and molecular weight. Specific antibodies directed against dystrophin, i.e., DRP2 alpha-dystrobrevin, beta-dystroglycan, alpha-syntrophin, alpha-, beta-, gamma-, and delta-sarcoglycan, and sarcospan, were used. The immunofluorescence study (confocal microscopy) showed differences in positive immunoreactions at the sarcolemmal membrane in these slow-type and fast-type skeletal muscle fibers. Protein extracts from T. marmorata red and white muscles were analyzed by Western blotting and confirmed the presence of dystrophin and associated proteins at the expected molecular weights. Differences were confirmed by comparative immunoprecipitation analysis of enriched membrane preparations with anti-beta-dystroglycan polyclonal antibody. These experiments revealed clear complex or non-complex formation between members of the dystrophin system, depending on the muscle type analyzed. Differences in the potential function of these various dystrophin complexes in fast or slow muscle fibers are discussed in relation to previous data obtained in corresponding mammalian tissues. (J Histochem Cytochem 49:857-865, 2001)

Immunohistochemical localization of protein gene product 9.5, ubiquitin, and neuropeptide Y immunoreactivities in epithelial and neuroendocrine cells from normal and hyperplastic human prostate.

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)

Estrogen receptors alpha and beta in the normal, hyperplastic and carcinomatous human prostate.

Two different estrogen receptors (ER-alpha and ER-beta) have been described, which are differentially involved in regulating the normal function of reproductive tissues. ER-alpha was considered for a long time to be the only estrogen receptor, and it has been detected in the stromal cells of the human prostate but not in the epithelium. To obtain new information about the differential effects of both receptor types, we have investigated their localization in normal prostates, benign prostatic hyperplasia (BPH), and prostatic cancer (PC) by immunohistochemistry, ELISA and Western blot. Epithelial immunostaining was absent in normal prostates and was present in BPH (10% of cells) and PC (80% of cells), whereas about 15% of stromal cells were positively immunostained for ER-alpha in the three types of prostatic specimens studied. Epithelial immunostaining for ER-beta was detected in normal prostates (13% of cells), BPH (30% of cells) and PC (79% of cells), whereas stromal immunostaining for ER-beta was absent in normal and hyperplastic prostates and was present in PC (12% of cells). The complementary presence of both receptor types in the normal prostate (ER-beta in the epithelium and ER-alpha in the stroma) might explain the mechanism of estrogen action in the development of BPH. The increased epithelial immunostaining for both ER-alpha and ER-beta in BPH and PC suggests that the involvement of estrogen receptors in hyperplasia and cancer concerns mainly the epithelium.