Valor pronóstico de la citología urinaria en la recidiva del carcinoma urotelial vesical tras la cistectomía radical / Prognostic value of urinary cytology for detecting urothelial carcinoma recurrence after radical cystectomy

Introducción: Las recidivas del carcinoma urotelial (CaU) en uretra o en el tracto urinario superior (TUS), tras una cistectomía radical (CR) son infrecuentes (4-6%), y su diagnóstico suele ocurrir en los 2 primeros años. Actualmente, no existen claras recomendaciones para la detección de recidivas en el urotelio remanente (UR), aunque se sabe que su detección precoz ofrece beneficios en la supervivencia. Nuestro objetivo es determinar el valor diagnóstico de la citología urinaria (CU) para la detección de recidivas en el UR y calcular su impacto como método de diagnóstico precoz en la supervivencia.Material y métodosRevisión retrospectiva de pacientes intervenidos de CR por CaU entre 2008-2016, con un seguimiento mayor de 24 meses.ResultadosSe incluyeron 142 pacientes. En una mediana de seguimiento de 68,5 meses, 9 pacientes (6,3%) presentaron recidivas en el UR (uretra: 4, TUS: 4, sincrónica: uno). La sensibilidad de la CU para el diagnóstico de recidivas en el TUS fue del 20% y la especificidad del 96%. No se encontraron diferencias significativas entre la supervivencia global y la supervivencia cáncer específica entre pacientes según el resultado de la CU.ConclusiónLas recidivas en el UR tras una CR son infrecuentes, y en nuestro estudio, hemos encontrado una baja sensibilidad para el diagnóstico de estas con CU. Por estas razones, no consideramos que la CU aporta información útil para el seguimiento de estos pacientes. (AU)

Introduction: Urethral or upper urinary tract (UUT) recurrence of urothelial carcinoma (UC) after radical cystectomy (RC) are rare (4-6%), and their diagnosis usually occurs within the first two years. Although it is known that its early detection offers benefit in terms of survival, currently there are no clear recommendations for the detection of recurrence in the remnant urothelium (RU). Our aim is to determine the diagnostic value of urinary cytology for the detection of recurrences in the RU and to estimate its impact as an early diagnostic method on survival.Material and methodsRetrospective review of patients who underwent RC for urothelial carcinoma between 2008-2016, with a follow-up of at least 24 months.ResultsThe study included 142 patients. In a median follow-up of 68.5 months, nine patients (6.3%) presented recurrences in the RU (urethra: four, UUT: four, synchronous: one). The sensitivity and specificity of urinary cytology for the diagnosis of UUT recurrences were 20% and 96%, respectively. No significant differences were found between overall survival and cancer-specific survival among patients according to the urinary cytology results.ConclusionRecurrences in the RU after RC are infrequent; our study has shown that urinary cytology offers a low sensitivity for their diagnoses. For these reasons, we do not consider that urinary cytology provides useful information for surveillance of these patients. (AU)

Prognostic value of urinary cytology for detecting urothelial carcinoma recurrence after radical cystectomy.

INTRODUCTION: Urethral or upper urinary tract (UUT) recurrence of urothelial carcinoma (UC) after radical cystectomy (RC) are rare (4-6%), and their diagnosis usually occurs within the first two years. Although it is known that its early detection offers benefit in terms of survival, currently there are no clear recommendations for the detection of recurrence in the remnant urothelium (RU). Our aim is to determine the diagnostic value of urinary cytology for the detection of recurrences in the RU and to estimate its impact as an early diagnostic method on survival. MATERIAL AND METHODS: Retrospective review of patients who underwent RC for urothelial carcinoma between 2008-2016, with a follow-up of at least 24 months. RESULTS: The study included 142 patients. In a median follow-up of 68.5 months, nine patients (6.3%) presented recurrences in the RU (urethra: four, UUT: four, synchronous: one). The sensitivity and specificity of urinary cytology for the diagnosis of UUT recurrences were 20% and 96%, respectively. No significant differences were found between overall survival and cancer-specific survival among patients according to the urinary cytology results. CONCLUSION: Recurrences in the RU after RC are infrequent; our study has shown that urinary cytology offers a low sensitivity for their diagnoses. For these reasons, we do not consider that urinary cytology provides useful information for surveillance of these patients.

[Prostate specific antigen and NF-kB in prostatic disease: relation with malignancy]. / Antígeno prostático específico y NF-kB en patología prostática: relación con la malignidad.

INTRODUCTION: NF-kB (p50/p65) is a transcription factor involved in TNF-α-induced cell death resistance by promoting several antiapoptotic genes. We intend to relate the expression of NF-kB (p50 and p65) with serum levels of prostate-specific antigen (PSA), both in normal males and in those with pathologic conditions of the prostate. MATERIALS AND METHODS: this study was carried out in 5 normal, 24 benign prostatic hyperplastic (BPH) and 19 patients with prostate cancer (PC). Immunohistochemical and Western blot analyses were performed on tissue and serum PSA was assayed by PSA DPC Immulite assays (Diagnostics Products Corporation, Los Angeles, CA). RESULTS: in controls, p65 NF-kB was not found and p50 was scantly detected in 60% normal samples in the cytoplasm of epithelial cells. Both p50 and p65 were expressed in 62.5% of the samples with BPH and in 63.2% of those with PC. Both increased its frequency of expression with higher PSA serum levels. CONCLUSIONS: Activation of NF-kB revealed by its nuclear translocation in prostate cancer could be related to cancer progression and elevated seric PSA levels. A better understanding of the biologic mechanism by which circulating PSA levels increase and its relation with NF-kB expression is needed. Possibly, NF-kB blockage could be used as a therapeutic target to counteract proliferation in prostate cancer.

Antígeno prostático específico y NF-kB en patología prostática: relación con la malignidad / Prostate specific antigen and NF-kB in prostatic disease: relation with malignancy

Introducción: NF-kB (p50/p65) es un factor de transcripción implicado en la resistencia a muerte celular provocada por TNF-α que promueve diferentes genes antiapoptóticos. Pretendemos relacionar la expresión de NF-kB con los niveles de antígeno prostático específico (PSA) en suero, tanto en varones sanos como en los que padecen condiciones patológicas de la glándula próstatica. Métodos: el estudio se realizó en 5 varones sanos (controles), 24 pacientes con hiperplasia benigna de próstata (HBP) y 19 pacientes con cáncer de próstata (CP). Se llevó a cabo Western blot e inmunocitoquímica en tejido y se evaluó el PSA sérico mediante PSA DPC immulite assays (Diagnostics Products Corporation, Los Ángeles, CA). Resultados: en los controles no se detectó el componente p65 de NF-kB y el p50 se detectó débilmente en el 60% de las muestras en el citoplasma de células epiteliales. Tanto p50 como p65 se expresaron en el 62,5% de las muestras de HPB y en el 63,2% de los pacientes con CP. Ambos aumentaron su frecuencia de expresión a mayor nivel de PSA. Conclusiones: la activación de NF-kB puesta en evidencia por translocación nuclear en CP parece estar estrechamente relacionada con la progresión de la enfermedad y con los niveles séricos de PSA. Se necesita un mejor conocimiento del mecanismo biológico de la elevación del PSA circulante y de su relación con la expresión de NF-kB. Tal vez el bloqueo de NF-kB podría emplearse como diana terapéutica para frenar la proliferación del cáncer de próstata (AU)

Introduction: NF-kB (p50/p65) is a transcription factor involved in TNF-α-induced cell death resistance by promoting several antiapoptotic genes. We intend to relate the expression of NF-kB (p50 and p65) with serum levels of prostate-specific antigen (PSA), both in normal males and in those with pathologic conditions of the prostate. Materials and methods: this study was carried out in 5 normal, 24 benign prostatic hyperplastic (BPH) and 19 patients with prostate cancer (PC). Immunohistochemical and Western blot analyses were performed on tissue and serum PSA was assayed by PSA DPC Immulite assays (Diagnostics Products Corporation, Los Angeles, CA). Results: in controls, p65 NF-kB was not found and p50 was scantly detected in 60% normal samples in the cytoplasm of epithelial cells. Both p50 and p65 were expressed in 62.5% of the samples with BPH and in 63.2% of those with PC. Both increased its frequency of expression with higher PSA serum levels. Conclusions: Activation of NF-kB revealed by its nuclear translocation in prostate cancer could be related to cancer progression and elevated seric PSA levels. A better understanding of the biologic mechanism by which circulating PSA levels increase and its relation with NF-kB expression is needed. Possibly, NF-kB blockage could be used as a therapeutic target to counteract proliferation in prostate cancer (AU)

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.

[Treatment of urinary stones with laser in special conditions]. / Tratamiento de la litiasis con láser en condiciones especiales.

During the last decade there has been a rapid development in flexible nephroscopy, flexible ureterorenoscopy, laser lithotripsy and instruments for stone manipulation. We are going to review the use of Laser in the management of lithiasis in different situations. Efforts should be made to minimize renal injury and lasers play a significant role in patients with urolithiasis and horseshoe kidneys, chronic renal failure, neurological patients.

The p38 transduction pathway in prostatic neoplasia.

It has been proposed that, among other cellular responses, TNF-alpha induces not only cell death, but also cell proliferation by activation of p38. It has also been reported that IL-1-alpha favours cell proliferation by p38 activation. The aim of the present study was to evaluate upstream (alpha-PAK, MEK-6) and downstream (Elk-1 and ATF-2) components of the p38 transduction pathway in normal prostate, benign prostatic hyperplasia (BPH), and prostate carcinoma (PC). Immunohistochemical and western blot analyses were performed in 20 samples of normal prostate, 47 samples of BPH, and 27 samples of PC. In all normal prostates, immunoreactivity for p-Elk-1 and p-ATF-2 was observed in epithelial cell nuclei, but no expression of alpha-PAK or MEK-6. In BPH, there was expression of alpha-PAK (cytoplasm) and MEK-6 (cytoplasm), while the proportions of lesions that were immunoreactive for p-Elk-1 (nucleus and cytoplasm) and p-ATF-2 (nucleus) decreased. In PC, the percentages of cells that were immunoreactive for alpha-PAK (cytoplasm) or MEK-6 (cytoplasm) rose slightly in comparison with BPH, while the percentages of cells that were immunoreactive for p-Elk-1 (nucleus and cytoplasm) or p-ATF-2 (nucleus and cytoplasm) were much higher than in BPH. It is concluded that overexpression of alpha-PAK, MEK-6, p38, p-Elk-1, and p-ATF-2 in BPH, and more intensely in PC, enhances cell proliferation. In BPH, such proliferation is triggered by IL-1 and in part counteracted by the TNF-alpha/AP-1 pathway, which promotes apoptosis. In PC, proliferation is triggered by IL-1 and TNF-alpha (the TNF-alpha/AP-1 pathway is diverted towards p38 activation). Since in a study of the same patients immunoexpression of IL-1alpha and IL-1RI was previously observed to be increased in PC, inhibition of p38 is a possible target for PC treatment, as this inhibition would both decrease IL-1-induced cell proliferation and increase TNF-alpha-induced cell death.

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.

Immunoexpressions of p21, Rb, mcl-1 and bad gene products in normal, hyperplastic and carcinomatous human prostates.

A comparative study of the expression of p21, Rb, mcl-1, and bad gene products, which are involved in the control of the cell cycle, was performed in normal, hyperplastic, and carcinomatous human prostates by means of a semiquantitative immunochemical study. This included Western blot, ELISA, and immunohistochemistry procedures. In normal prostates, immunoexpression of the four gene products was scanty or absent. In men with benign prostatic hyperplasia, immunoreactions to the four proteins studied were found in many epithelial cells and some stromal cells. In prostatic carcinoma, the immunostaining pattern was as in hyperplastic prostates but the numbers of both epithelial and stromal cells were higher. Present results indicate that immunoexpression of p21, Rb (both the phosphorylated and dephosphorylated forms), mcl-1, and bad gene products are markedly increased in prostates with proliferative alterations but that these proteins do not discriminate between benignant (hyperplasia) and malignant (adenocarcinoma) prostatic tumours, although immunoexpression is higher in prostatic carcinoma.

IL-2, its receptors, and bcl-2 and bax genes in normal, hyperplastic and carcinomatous human prostates: immunohistochemical comparative analysis.

The presence of interleukin-2 (IL-2) and its receptors (Ralpha, Rbeta, Rgamma), and their relationship with the products of bcl-2 and bax genes was studied in normal prostates, benign prostatic hyperplasia (BPH), and prostatic cancer (PC) by ELISA, Western blot, and immunohistochemistry. A comparative semiquantitative immunohistochemical study was also performed. For all the antibodies assayed, immunoreactions were found in the epithelium and some stromal cells in the three types of specimens studied. These immunoreactions were much more higher in PC samples than in normal prostates. In BPH, immunoreactions were similar to that of normal prostates (bax), similar to that of PC (IL-2 and its three receptors), or intermediate between that of normal prostates and that of PC (bcl-2). Immunoexpressions of IL-2 and its receptors were found in the epithelial basal cells and some stromal cell of normal prostates and might be related to the control of the proliferation-apoptosis equilibrium. The increased expressions of IL-2 and its receptors in the epithelium of prostates in BPH, associated with increased bcl-2 expression which would account for the decrease in the apoptosis index that has been reported in this disorder. The increased expression of both bcl-2 and bax in PC might be involved in the higher apoptosis index reported in PC specimens. Since IL-2 administration seems to have an anti-tumour effect, the increased expression of this interleukin in BPH and PC could be interpreted as an attempt to hinder cell proliferation which would only be efficient at high doses.

Immunoexpression of tumour necrosis factor-alpha and its receptors 1 and 2 correlates with proliferation/apoptosis equilibrium in normal, hyperplasic and carcinomatous human prostate.

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.

A lectin histochemistry comparative study in human normal prostate, benign prostatic hyperplasia, and prostatic carcinoma.

The partial oligosaccharide sequences of glycoconjugates and the nature of their glycosidic linkages were investigated in normal human prostate, benign prostatic hyperplasia (BPH) and prostatic carcinoma by means of lectin histochemistry, using light microscopy and Western blot analysis. The labeling pattern of BPH differed from that of normal prostate in having more intense staining with DSA, HPA, UEA-I and AAA, and in showing lesser staining with WGA and SBA. Prostatic carcinoma differed from normal prostates in displaying the more intense labeling with PNA, DSA, SBA, DBA, UEA-I and AAA, and in having lesser labeling with WGA. The main differences in labeling pattern between prostatic carcinoma and BPH were that the latter specimens showed more marked staining with PNA, DSA, DBA, SBA, UEA-I and AAA, and lesser staining with WGA and HPA. The staining patterns of SNA, MAA, ConA, LCA and GNA were similar in all three groups of specimens. For most of the lectins studied, including those showing a similar immunohistochemical staining in the three groups of specimens studied, the Western blot analysis showed differences in the banding pattern among normal, hyperplastic, and carcinomatous prostates. Present results suggest that the glycosylation of proteins was modified in both BPH and prostatic carcinoma. In BPH a strong expression of N-acetylgalactosamine residues occurred, while in prostatic carcinoma an increase of sialic acid, galactose and fucose residues was observed. No changes in mannose residues were detected.

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.

Total and regional bone mass and biochemical markers of bone remodeling in metastatic prostate cancer.

BACKGROUND: The osteolytic activity of metastases of prostate cancer was evaluated in relation to total body bone mineral content (TBBMC) and regional bone mineral content (RBMC). METHODS: Bone mass was determined by dual-energy X-ray absorptiometry (DXA). Tartrate-resistant acid phosphatase (TRAP) was measured as a biochemical marker of bone resorption. RESULTS: In 32 patients (mean age 72+/-4 years) compared with 32 controls (mean age 73+/-5 years), there were significant differences in TRAP (P < 0.0001), TBBMC (P < 0.0001), and RBMC in the pelvis (P < 0.0001), legs (P=0.0001), and trunk (P<0.05), but not in the arms and head (P=ns). In the overall group of subjects, the correlation between TBBMC and TRAP was r=-0.68, P < 0.0001. The correlations remained significant in the patient and control groups separately. CONCLUSIONS: The loss of bone mass observed in patients with metastatic prostate cancer was caused mainly by the predominance of bone resorption in the osteoblastic metastases.

Transforming growth factor beta 1 and its receptor types I and II. Comparison in human normal prostate, benign prostatic hyperplasia, and prostatic carcinoma.

An immunohistochemical and semiquantitative comparative study of transforming growth factor beta 1 (TGF-beta 1) and its receptor types I (TGF-beta RI) and II (TGF-beta RII) was carried out in normal prostates and in the prostates from men with benign prostatic hyperplasia (BPH), and men with prostatic adenocarcinoma. Immunoreaction to TGF-beta 1 was limited to the basal epithelial cells in the normal prostates. Some cells in the connective tissue stroma were also stained. In BPH immunolabelling was also observed in columnar (secretory) cells of the epithelium. In prostatic adenocarcinoma, all epithelial cell types were intensely immunostained. Some stromal cells were also stained. Immunostaining to TGF-beta RI was only present in the basal cells in normal prostates. In BPH, this immunoreaction was found in the whole epithelium and in some stromal cells. In prostatic cancer, the immunostaining pattern for this receptor was similar to that of BPH but more intense in the epithelial cells. Immunoreactivity to TGF-beta RII appeared in some basal cells and some scattered columnar cells of the normal prostate epithelium. In the BPH sections, this pattern was maintained, and a weak immunolabelling was also observed in the stroma. In prostate cancer, all epithelial cells appeared intensely labelled. In the stroma, immunolabelling was similar to that of the BPH specimens. The results of the present study suggest that, in normal prostates, only the basal cells of the epithelium possess both receptor types, and hence can transduce TGF-beta 1 signal intracellularly. The basal cells can also secrete this growth factor which would act as an autocrine inhibitory growth factor for them. In addition, TGF-beta 1 is secreted in some zones by stromal cells, acting then as a paracrine growth factor for basal cells in those areas. In BPH, in addition to the basal cells, some secretory columnar cells also secrete TGF-beta 1 and possess both types of TGF-beta 1 receptors, and thus, both epithelial cell types are susceptible to TGF-beta 1 action. Since both receptor types are also present in some stromal cells, these cells also perform an autocrine secretion, in addition to their paracrine secretion to the epithelial cells. TGF-beta RIIs seem to be more numerous than TGF-beta RIs and this lead us to hypothesize that these incomplete receptors might be a protection against the inhibition caused by TGF-beta 1 action. In prostatic carcinoma all cell types display the same characteristics as in BPH, although both receptor types are found in similar numbers, and thus, the above mentioned protection would not occur.

Lectin histochemistry of the human testis.

The oligosaccharide sequences of glycoconjugates and the nature of the saccharide linkage were investigated in normal human testes by means of lectin histochemistry studies, at light and electron microscopy levels. Reaction to WGA was intense in the seminiferous epithelium and interstitium. MAA showed light reactivity in all cell types of the human seminiferous epithelium, the lamina propria and Leydig cells. UEA-I lectin labelled the lamina propria intensely and the seminiferous epithelium and Leydig cells slightly. A slight reaction to AAA was found in the seminiferous epithelium and in Leydig cells. ConA was labelled in Sertoli cells, germ cells and Leydig cells. The reaction to GNA lectin was similar although less intense. PNA labelling was slight in Sertoli cells, spermatogonia, and Leydig cells, and more intense in spermatocytes, spermatids and peritubular cells. Reaction to DSA was intense in the seminiferous epithelium and Leydig cells. HPA labelled all cell types in the seminiferous epithelium and Leydig cells slightly, and labelled peritubular cells intensely. SBA lectin showed a strong reaction in spermatids and a slight reaction in the lamina propria. The reactions to SNA, LTA, and DBA were negative in all testicular cell types. After beta-elimination pre-treatment, MAA, UEA-I, AAA, PNA, DSA, HPA and SBA reactions were all negative. Endo F/PNGase digestion suppressed reactivity to ConA y GNA. Staining for WGA decreased with Endo F/PNGase digestion and also after beta-elimination. Desialization increased reactivity to PNA, SBA and HPA lectins. These results indicate that the terminal sequences of oligosaccharide side-chains in spermatocytes and, principally, in spermatids are: fucose, mannose, Neu5Ac2,3Gal1,3GalNAc, Gal1,3GalNAc, Gal1,4GlcNAc, Neu5AcGalNAc and GalNAc (in O-glycosylated proteins); mannose (in N-glycosylated proteins) and GlcNAc (in both protein types). A sialic acid residue is added to galactose and GalNAc residues. Present findings also indicate that Sertoli cell glycoproteins are similar to those of spermatids, and that the terminal sugar residues in Leydig cells are GlcNAc, fucose, mannose, Neu5Ac2,3Gal1,3GalNAc, Gal1,3GalNAc, and Gal1,4GlcNAc. The lectin pattern of the lamina propria suggests the presence of GlcNAc, galactose, fucose and GalNAc.

Lectin histochemistry study in the human vas deferens.

The oligosaccharide sequences of glycoconjugates in the normal human vas deferens and the nature of the saccharide linkage were studied by lectin histochemistry. The cytoplasm of all epithelial cell types (principal cells, basal cells, and mitochondria-rich cells) and luminal contents reacted positively with WGA, MAA, PNA, DSA, LTA, UEA-I, AAA, and ConA. The reaction was more intense in the stereocilia of principal cells. Cytoplasmic staining was diffuse except for PNA and DSA labeling which was limited to the apical cytoplasm and stereocilia of columnar cells. The cytoplasm of all cell types also reacted diffusely with HPA, although staining was weak and was not observed in the stereocilia. Positive reaction with SBA only was encountered in the stereocilia of principal cells. SNA, LTA, and DBA were unreactive. GNA-labeling showed a granular distribution in the supranuclear cytoplasm of columnar epithelial cells. Reactions with MAA, PNA, DSA, AAA, HPA and SBA disappeared after the beta-elimination reaction. Reactions with WGA and UEA-I decreased after beta-elimination or Endo-F digestion. Reactions with ConA and GNA were suppressed by Endo-F digestion. Reactions with PNA, HPA, and SBA increased after desialylation. Of all the lectins that label the luminal contents of the vas deferens, only UEA-I was not found in the luminal contents of seminiferous tubules and epididymis and, thus, this lectin would probably bind to glycoproteins secreted by the vas deferens. The chemical treatments used suggest that this secretion contains fucose residues located in both N- and O-linked oligosaccharides. The other lectins may label secreted proteins, but also structural proteins or proteins reabsorbed from the luminal fluid. The lectin- binding pattern of mitochondria-rich cells in the vas deferens differed from that found in the epididymis.

[The Leydig-cell testicular tumor. A case report with magnetic resonance images]. / Tumor testicular de células de Leydig. Presentación de un caso con imágenes por resonancia magnética.

OBJECTIVE: An additional case of testicular Leydig cell tumor in a young man is reported. The clinical presentation included gynecomastia and a testicular mass. METHODS/RESULTS: Hormonal studies, testicular US and high resolution MRI were performed before orchidectomy. The histopathological findings were consistent with Leydig cell tumor. A CT scan revealed no metastasis. The patient is clinically disease-free 18 months after orchidectomy. CONCLUSION: High resolution MRI must be performed only when clinic and US findings are discordant.

Intermediate filaments in the Sertoli cells of the ageing human testis.

The presence and distribution of intermediate filaments (vimentin, keratin, desmin) was studied in the Sertoli cells of elderly men by means of quantitative immunohistochemical methods. Sertoli cells from young men showed moderate immunogold labelling to vimentin throughout the entire cytoplasm between the cell organelles in tubules showing complete spermatogenesis. Immunogold particles were more numerous in the perinuclear cytoplasm and beneath the plasma membrane in all its faces. The testes from elderly men showed different tubule types; some showed complete spermatogenesis and a normal lamina propria, while others had spermatogenic arrest at different levels (spermatids, spermatocytes, spermatogonia). The immunohistochemical reaction to vimentin in the Sertoli cells of tubules with complete spermatogenesis (type a) was similar to that in the cells of young men. In the Sertoli cells of severely damaged tubules (type b) the immunohistochemical reaction was more intense and immunogold particles extended in similar proportions throughout the whole cytoplasm. When immunolabelling intensity was compared between the three groups of tubules, by counting the number of immunogold particles per square micrometre of cytoplasm, it was found to be significantly higher (P < or = 0.05) in type b tubules of elderly men than either in tubules of young men or in type a tubules of elderly men. Since the average cell surface of Sertoli cells was similar in all tubule types, these data suggest that an actual vimentin increase occurs in Sertoli cells of germ-cell-depleted tubules. Sertoli cell immunogold labelling to keratin was found neither in young men nor in type a tubules of ageing men, whereas a positive immunohistochemical reaction was observed in the Sertoli cells of type b tubules of elderly men. Immunogold particles were localized mainly in the perinuclear cytoplasm, and beneath the lateral and basal cell surfaces. The observation of vimentin increase and keratin re-expression in ageing Sertoli cells only in germ-cell-depleted tubules suggests that the changes in intermediate filaments are related to the local factors associated with completion of spermatogenesis, causing functional changes in Sertoli cells.

Immunocytochemical and quantitative study of the tunica albuginea testis in young and ageing men.

A light and electron microscope immunohistochemical study of the tunica albuginea from both young and elderly men was carried out to determine the distribution of the cells that contain actin, vimentin and/or desmin, and to evaluate the possible variations with ageing by means of quantitative studies. Testicular volume and testicular parenchyma volume decreased significantly with age whereas the tunica albuginea volume remained unchanged. These results agree with the scanty quantitative changes observed in the testicular connective tissue with age, and the notion that age-related changes in testicular volume are principally restricted to the seminiferous tubules. Three connective tissue layers could be distinguished in the tunica albuginea in both young and elderly men. The middle and inner layers increased in width with age while the width of the outer layer decreased. The average width of the tunica albuginea increased significantly with ageing. The tunica albuginea of young men and elderly men presented two types of fusiform cells: (1) fibroblast-like cells, which immunoreacted to actin and vimentin, but not to desmin; and (2) myoid cells, which immunoreacted to actin, vimentin and desmin. In both young men and elderly men, the total number of desmin-positive cells (myoid cells) was significantly lower than that of fibroblasts. However, the total number of desmin-positive cells was significantly increased in ageing men. In young testes, desmin-positive cells were more abundant in the outer layer of the tunica albuginea, whereas in elderly men these cells predominated in the middle layer. The increased desmin immunoexpression in the tunica albuginea of ageing men contrasts with the decrease in desmin immunoreaction in other myoid cells of the testis, the peritubular myoid cells, but only in seminiferous tubules that showed severe germ cell depletion. This suggests that changes in intermediate filament immunoexpression in peritubular cells are focalised, and thus, under local control, whereas changes in the tunica albuginea cells are generalised and possibly related to factors also affecting the connective tissue in other organs.