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Aquaporin-9 (AQP-9) regulates tissue hydration by promoting transmembrane exchanges of both water and solutes, such as lactate. The latter is a key metabolite of primary spermatocytes and of maturing haploid germ cells (h-GCs). The present investigation was aimed at immunolocalising human AQP-9 in both normal and varicocele testes. Histology and immmunocytochemistry were investigated in archival biopsies from 20 varicocele testes and in eight unaffected ones. AQP-9 immunostaining was performed using a rabbit antibody, and either focal or diffuse cell membrane labelling was recorded. Varicocele testes showed disarranged tubular compartments, with sloughing h-GCs, tissue hyperhydration, spermiogenesis failure and fibrosis. AQP-9 immunohistology of the control testes showed a diffuse cell membrane staining of the primary spermatocytes and h-GCs, without any positive reaction of spermatogonia and Sertoli cells. AQP-9 cell expression in the varicocele testes was focal or lacking in both adluminal and sloughing GCs. AQP-9 expression occurs in normal human testis, at cell membrane of primary spermatocytes and h-GCs, suggesting a possible role of AQP-9 in the water and lactate transport from Sertoli cells to GCs. AQP-9 is focal or lacking in adolescent varicocele testes, and this suggests AQP-9 to be downregulated in such testicular disorder, leading to lactate deprivation with subsequent hypospermatogenesis., (© 2010 Blackwell Verlag GmbH.)

Introduction: Bladder augmentation predisposes humans to many metabolic, renal and enteropatch changes. Our aim was to evaluate in a rat model of ileocystoplasty mid- and long-term urinary, metabolic, renal and graft changes., Materials and Methods: We performed an ileocystoplasty and a sham operation in 30 rats. Seven augmented rats and 3 sham-operated animals were euthanized after 1, 3 and 6 months. We performed urinalysis, urine culture and blood sampling for serum electrolytes and urea. Histopathological changes of the ileal patch and kidneys were also evaluated., Results: The urine cultures were positive in 14.3, 57 and 71%, respectively, 1, 3 and 6 months after surgery. Urinary pH, serum chloride and urea of the augmented group were significantly higher. Bladder calculi were formed in 23.8% of ileocystoplasty. Histopathological examination showed urothelialization of the graft with hyperplastic/metaplastic changes. The kidneys showed glomerular depletion and a marked distal tubular ectasia., Conclusions: Our data showed that, in a mid- and long-term follow-up, rat bladders subjected to ileocystoplasty displayed hyperchloremic metabolic acidosis, electrolyte imbalance, enhanced serum blood urea levels and glomerular/tubular changes. Hyperplastic and/or metaplastic changes at the junctional zone were observed. Our experimental results suggest that frequent monitoring of renal function and surveillance of humans who have undergone ileocystoplasty are recommended., (Copyright © 2011 S. Karger AG, Basel.)

Objective: To immunolocate c-kit-positive interstitial cells of Cajal (ICCs, known to be responsible for pacemaker activity in human ureters, coordinating ureteric motility) in the intramural ureter of patients with different grades of vesico-ureteric reflux (VUR), to assess the ureteric histology and correlate these findings with manometric patterns., Patients and Methods: The VU junction (VUJ) represents the boundary between the low-pressure of the upper and high pressure of the lower urinary tract, protecting the upper tract from VUR using active and passive antireflux mechanisms. The contraction of the longitudinal muscle coat of the VUJ possibly functions as an 'active' antireflux system, but previous manometric findings on refluxing ureteric units (RUs) have shown altered patterns. In all, 32 RU ends were stained using both picro-Mallory and Sirius Red techniques; in a parallel immunohistochemical procedure, using mast cell tryptase and CD117 antibodies (to identify ICCs), they were compared with eight control ureteric ends. Ureteric manometry of the VUJ was also done during ureteric reimplantation., Results: The histochemical and immunohistochemical results in the RUs showed a replacement of the altered smooth muscle fascicles by collagenous stroma and significant loss of ICCs in RU ends, both correlated with the grade of VUR. Ureteric manometry showed significant impairment of basal and maximum pressure in RUs, correlated, respectively, with histological lesions and loss of ICCs., Conclusion: Deficiency of the longitudinal muscle coat probably leads to dysfunction and insufficiency of the ostial valve mechanism, with subsequent impairment of the active valve mechanism. Histological, histochemical and immunohistochemical changes support the alterations of ureteric peristalsis in RUs.

Background: Aberrant Wingless type 1 glycoprotein (Wnt) pathway in ameloblastomas and a role of syndecan-1 (SDC1) in activating Wnt signalling were perspected. SDC1 shifting from epithelium to stroma was reported in invasive non-odontogenic neoplasms. The aim of this study was to reveal the role of SDC1 and Wnt1 in intraosseous ameloblastomas (IA(s))., Methods: SDC1 and Wnt1 expressions were investigated in 29 ameloblastoma subtypes and seven tooth buds., Results: SDC1 immunostaining strongly depicted stromal cells, extracellular matrix (ECM) and basement membranes of ameloblastomas. It also showed epithelial tumour cells in the acanthomatous and plexiform subtypes, and it often occurred in stellate reticulum cells and basal ameloblasts of tooth buds. Parallel Wnt1 expression occurred in ameloblastomatous epithelial cells, but it was common in basal cells of tooth buds too. Statistically, a significant correlation was found between the percentage of IA(s)-bearing SDC1-positive stromal cells and ECM and the percentage of IA(s)-bearing Wnt1-positive epithelial cells., Conclusions: A role of SDC1 in stromal cells and ECM can be hypothesized as a critical factor for carcinogenesis and local invasiveness of IA(s).

Aim: Recent reports have shown an altered density of interstitial cells of Cajal (ICCs) and peripherin immunoreactive nerve fibres in uretopelvic junction obstruction. The aim was to investigate ICCs immunoexpression and ureteral innervation in primary obstructive megaureter (POM)., Methods: Eight specimens of POM were obtained during tailoring. Restricted segments of ureters were divided from dilated segments. C-kit and peripherin immunohistochemistry were performed., Results: A normal distribution of ICCs was observed in both the circular and longitudinal muscle layers of the dilated segments. Marked muscle hypoplasia and sparse or no ICCs occurred in the longitudinal muscle layer of the restricted ureteral segments. A normal distribution of peripherin positive fibres was present in both the dilated and restricted segments., Conclusions: Our data confirm defective muscularization in restricted aperistaltic POM. The lack of ICCs in the longitudinal muscular layer is probably due to the absence of c-kit positive muscle embryological precursors. No alteration in the peripherin immunoreactive nerve fibres network was observed in either the dilated or the restricted ureteral segments.

Objectives: Recent reports showed an altered density of interstitial cells of Cajal (ICCs) and of peripherin immunoreactive nerve fibres in obstructed uretero-pelvic junction. Our aim was to investigate ICCs immunoexpression and ureteral innervation in primary obstructive megaureter (POM)., Methods: 8 specimens of POM were obtained during tailoring. Restricted segments of ureters were divided from the dilated ones. C-kit and peripherin immunohistochemistry were performed., Results: A normal distribution of ICCs was documented in both circular and longitudinal muscular layers of dilated segments. Marked muscular hypoplasia and a sparse or absent ICCs occurred in longitudinal muscular layer of restricted ureteral segments. A normal distribution of peripherin positive fibres was present in both dilated and restricted segments., Conclusions: Our data confirmed a defective muscolarization in restricted, aperistaltic POM. The lacking of ICCs in the longitudinal muscular layer is, probably, due to the absence of c-kit positive muscular embryological precursors. No alteration of peripherin immunoreactive nerve fibres network was observed in both dilated and restricted ureteral segments.

Endometrial polypoid adenomyomatosis in an 8-year-old German shepherd bitch is described. The lesion was associated with ovarian granulosa cell tumour and pyometra; grossly, it consisted of sessile or pedunculated processes with both epithelial and non-epithelial components, in which smooth muscle cells were predominant. The endometrium was diffusely atrophic and showed multifocal squamous metaplasia. The findings are discussed as possible consequences of the functioning ovarian tumour and pyometra, but an involvement of growth factors is also proposed.

Purpose: Testicular damage after torsion has been attributed to many mechanisms, of which one is lipid peroxidation of the plasma membrane, which could cause the activation of the mitogen-activated protein kinase family. These proteins are of vital importance for signal transduction pathways and 2 of them, extracellular signal-regulated kinase and c-jun N-terminal kinase, participate in the pathogenesis of testicular ischemia. We investigated whether lipid peroxidation may trigger mitogen-activated protein kinase activation in testicular ischemia-reperfusion., Materials and Methods: Adult male Sprague-Dawley rats were subjected to 1-hour testicular ischemia, followed by 24 hours of reperfusion. Sham testicular ischemia-reperfusion rats served as controls. Animals were randomized to receive raxofelast, an inhibitor of lipid peroxidation (20 mg/kg intraperitoneally administered 15 minutes before detorsion and 15 minutes after detorsion) or vehicle (1 ml/kg 10% dimethyl sulfoxide/NaCl solution). A group of animals was sacrificed 0, 10, 15, 20, 25 and 30 minutes, and 1, 2 and 3 hours, respectively, after detorsion to evaluate testicular c-jun N-terminal kinase, extracellular signal-regulated kinase and tumor necrosis factor-alpha activation by Western blot analysis, and mRNA expression and conjugated dienes using a spectrophotometer technique. Another group was sacrificed 24 hours after detorsion to evaluate histological alterations., Results: Testicular ischemia-reperfusion injury caused a significant increase in the conjugated diene levels, extracellular signal-regulated kinase c-jun N-terminal kinase activity and tumor necrosis factor-alpha expression in both testes. Furthermore, histological examination revealed marked damage. Raxofelast inhibited these parameters and decreased histological damage., Conclusions: These data suggest that lipid peroxidation triggers extracellular signal-regulated kinase and c-jun N-terminal kinase activation. Furthermore, mitogen-activated protein kinase blockade might represent a potential therapeutic approach to treatment in patients with unilateral testicular torsion.

Aim: To highlight the ultrasonographic features of prenatal torsion of the testis in utero (IUTT) at presentation, the neonatal management and the histological findings postorchiectomy or biopsy., Methods: Seven newborns underwent emergency exploration for IUTT. All patients underwent a sonography and real-time color Doppler ultrasound study of the scrotum before any surgical procedure. A histological examination was performed in the removed specimens., Results: Sonography of the scrotum revealed enlarged, heterogeneous testes. In all cases the color and power Doppler did not reveal any flow signal on the affected side. Four newborn with unilateral testicular torsion underwent orchiectomy and contralateral orchidopexy. In one neonate after detorsion and with the absence of gangrenous changes and a reassuring biopsy, a twisted testis could be treated conservatively with orchidopexy. In another case, the parents, acknowledging the inviability of the affected testis, gave consent only for a biopsy of the testis. In the neonate with bilateral IUTT, bilateral testicular biopsies were performed. Histology of the removed testes variably showed interstitial red cell extravasion and coagulation or hemorrhagic necrosis. Light microscopy of the preserved testis highlighted surviving seminiferous tubules, with gonocytes, spermatogonia and fetal Sertoli cells., Conclusions: An early diagnosis and treatment in IUTT is essential. Surgical exploration should be always performed through the inguinal route. In bilateral IUTT testes should be left to try to assure, as long as possible, a residual Leydig cell function.

Mitogen-activated protein kinase (MAPK) 3/MAPK1 (also known as ERK1/ERK2) plays an important role in the signal transduction pathways. To our knowledge, however, its role in the development of testicular ischemia-reperfusion injury has not yet been investigated. Therefore, we studied the pattern of MAPK3/MAPK1 activation in a experimental model of testicular ischemia-reperfusion injury. We also investigated MAPK8 to understand whether an association exists between these two MAPKs. Adult male Sprague-Dawley rats were subjected to 1 h of testicular ischemia followed by 24 h of reperfusion or to a sham testicular ischemia-reperfusion. Animals were randomized to receive PD98059, which is an inhibitor of MAPK3/MAPK1 (10 mg/kg i.p. administered immediately after detorsion), or its vehicle. The time course of MAPK3/MAPK1, MAPK8, and tumor necrosis factor (TNF; also known as TNF alpha) expression and a histological examination in both the ischemic-reperfused testis and the contralateral one were performed. In both testes, MAPK3/MAPK1 and MAPK8 expression appeared following 10 min of reperfusion and reached their highest activation after 30 min. The MAPK levels slowly decreased, and no significant expression of either kinase was observed following 2 h of reperfusion. Expression of TNF was evident after 1 h of reperfusion and reached its maximum increase after 3 h. PD98059 blunted MAPK3/MAPK1 and MAPK8, reduced TNF expression, and improved the testicular damage caused by ischemia-reperfusion injury in both testes. These data emphasize that MAPK3/MAPK1 has a role in testicular damage and that its blockade might have a future therapeutic role for the management of patients with unilateral testicular torsion.

Objectives: The aim of this study was to demonstrate the ultrasonographic features of prenatal bilateral torsion of the testis, and its histological correlation and management., Patient: A newborn presented at delivery with both testes enlarged, swollen and tender. Prenatal ultrasound (US) showed enlarged, hyperechoic testes. Colour Doppler US examination was performed., Results: US revealed both testes to be heterogeneous. Colour Doppler US did not reveal any flow signal. On inguinal exploration both testes appeared necrotic. Histology showed recognizable seminiferous tubules and Leydig cells., Conclusion: We believe that both testes should be left in situ after bilateral detorsion even if their macroscopic appearance is necrotic.

Objectives: To investigate the immunoexpression of aquaporin-1 (AQP-1), a major transmembrane water channel, in high-grade varicocele testes, which are known to imply an unbalanced transmembrane water flow, in both tubular and extratubular compartments., Methods: Light microscopy and AQP-1 immunohistologic examination were carried out on incisional testicular biopsies from 20 adolescent boys, aged 13 to 18 years, with grade 2 or 3 idiopathic varicocele. Testicular tissue from 4 autopsied subjects of matched age were also investigated as parallel controls., Results: At microscopic examination, the tubular lumina and extracellular matrix were expanded and venular profiles dilated. Cytoplasmic vacuoles were frequent in the Sertoli cells, spermatogonia, and spermatocytes, together with premature sloughing of germ cells and residual cytoplasmic droplets inside the spermatozoa, if present. Diffuse AQP-1 positivity occurred at venular endothelial cell membranes and, unexpectedly, at the cell membranes of the Sertoli cells, diploid germ cells, and haploid cells. In the control testes, focal AQP-1 immunolabeling was confined to the microvessel endothelial cells, without any positive reaction in the tubular or extratubular compartments., Conclusions: This is the first report showing AQP-1 cell expression in adolescent varicocele testes in both tubular and extratubular compartments. It was associated with the main histologic features of unbalanced water flow in the tubular and interstitial compartments of the testis. Immunocytochemical patterns revealed AQP-1 as a possible critical reabsorption factor, acting to reduce abnormal fluid retention in endotubular cells and the extracellular matrix and, to a lesser extent, in Leydig cells.

A 30-year-old Standardbred stallion was examined for unilateral scrotal swelling. Physical and ultrasound examinations revealed a painless enlarged left testis with a non-homogeneous echogenicity, when compared with the controlateral testis. The stallion underwent left unilateral orchiectomy. Grossly, the excised testis was irregularly enlarged (12 x 9 x 9 cm; weight: 530 g) and firm. The sections showed that testicular parenchyma was replaced by a lobulated, greyish-white mass, which involved the epididymal head. At microscopy, a dual Leydig and Sertoli cell tumour component could be seen. Neoplastic Sertoli cells were prevalent and presented pleomorphic cells, mitotic figures and occasional vascular invasion. Tumour patterns showed tubular and solid areas, cord-like or diffuse in appearance, among which newly formed Leydig cell nests and low-density fibrillar bundles were interposed. Immunohistochemically, a weak to moderate immunostaining for vimentin, AE(1)/AE(3) cytokeratin, alpha-1-antitrypsin and CD99 antigens was found in the growing Sertoli cells, whose nuclear MIB-1 labelling index scored 13 +/- 2%. The Leydig tumour cells, on the other hand, displayed a moderate to strong positivity for alpha-inhibin, vimentin, AE(1)/AE(3) cytokeratin, neurone-specific enolase and CD99. On the basis of these findings, a diagnosis of malignant mixed sex cord-stromal tumour was made.

Objectives: Both aquaporin (AQP) 1 and the stem-cell factor/C-kit system seem to have a definite role in testis function, but very few studies have been reported in humans, especially in the paediatric age group. With the present study we wanted to investigate the expression of these proteins to better delineate their role in normal and pathologic testes., Methods: Immunohistology using AQP 1 and C-kit antibodies was performed on paraffin sections of open-testicular biopsies from 32 undescended testes. The testes of cryptorchid patients, with ages ranging from 2 to 15 years, were biopsied during an orchidopexy operation, after obtaining informed consent. Control biopsies, from 8 patients of matched age, were obtained during operations for inguinal hernia or hydrocele, always after obtaining informed consent. Positive results were recorded as diffuse or focal patterns and scored as weak, moderate or strong immunostaining., Results: AQP 1 antibody strongly depicted microvessel endothelial cells, but was unlabeled in endotubular and interstitial cell lines, in both control and undescended testes. The C-kit immunostaining in normal testes revealed a diffuse, strong staining in the cytoplasm of spermatogonia and primary spermatocytes. However, in the undescended testes a focal C-kit immunolabelling was weakly recognized in both spermatogonial and immature Sertoli cells., Conclusions: These results indicate a direct involvement of AQP 1 in the regulation of fluid transport across the endothelial cell membranes of testicular microvessels. A role of the C-kit receptor protein is also substantiated by its strong expression in the maturing spermatogonia of the normal testes, but was minimally or not recognizable in undescended prepubertal testes.

Objective: A case of Stage IIA primary ovarian leiomyosarcoma (LMS) with an unfavorable outcome 24 months after total abdominal hysterectomy with bilateral salpingo-oophorectomy, despite chemotherapy treatment, is described. Eighteen months from surgery the patient showed peritoneal spreading with ascites, liver and lung metastases. The present study was aimed to compare tumor growth fraction with cell density, lesion stage and clinical course., Methods: The surgical specimens were evaluated by histological, histochemical, and immunocytochemical methods. Under microscopy, mitotic index (MI) was estimated, as a ratio of mitotic figures per 1000 tumor cells. Immunohistology was also carried out to reveal some intermediate-type filamentous proteins, as histogenetic markers, and the MIB1 monoclonal antibody was used to assess the percent of MIB1-positive nuclei (MIB1 labeling index)., Results: The histologic findings and immunohistology of the assayed intermediate filamentous proteins substantiated a diagnosis of LMS, with associated coagulation necrosis and not rare mitotic figures. A dual tumor component was observed, representing both the pleomorphic and myxoid LMS-variants. On the basis of the quantitative findings, a MI of 10.9 and a MIB1-LI of 23.1 were calculated, on average., Conclusions: The proliferation indices in the described variant of ovarian LMS, denote a fast growing malignancy. They agree with the tumor stage at operation and the subsequent fatal outcome.

A homologous malignant mixed Müllerian tumor of the uterus occurring in an 8-year-old Persian cat was described with regard to its clinical and pathologic features. A polypoid multinodular mass of the right uterine horn was shown by an ultrasound examination. Grossly, the right uterine horn was enlarged because of a vegetative and infiltrating tumor, grayish-white in color, that penetrated the uterine wall to the level of the perimetrium. Many metastatic nodules were found in abdominal and thoracic cavities. Histologically, the neoplasm had both carcinomatous and sarcomatous components and was diagnosed as an uterine malignant mixed Müllerian tumor. This is the fourth case reported in cats. The histologic features and proliferation rate of this tumor were similar to the corresponding human neoplasms, which occur mainly in postmenopausal women. The possible hormone dependence of the tumor is briefly discussed.

Inhibin B immunocytochemistry of both the alpha- and beta-subunits was studied in testicular biopsies from 18 prepubertal and postpubertal patients, with unilateral or bilateral cryptorchism. The present investigation was carried out to seek any prognostic significance for the expected fertility of such subjects in adulthood. All samples were also evaluated by histological and morphometric assessments, according a 1-6 grading sy-stem. In this way, the individual testicular changes were scored by quantitating tubular and germ cell hypoplasia, Sertoli cell hyperplasia, and peritubular fibrosis, where present. The results showed that in bilateral maldescended testes an unexpected expression often occurred of inhibin B beta-subunit in Sertoli cells, while inhibin B alpha-subunit there did not, denoting an early developmental arrest of the testis. It co-related with the high grade testicular damages, as a poor predictor of spermatogenesis. Unlike, unilateral retained testes mainly expressed inhibin B alpha-subunit, irrespective of tubular changes. In the latter instance, different pathogenetic factors of imbalanced testicular regulation can be perspected, other than the Inhibin-Activin system.

A case of uterine cholesterol granuloma in a 12-year old mixed breed cat is reported. The lesions were found in the endometrium of the left uterine horn as scattered, raised nodules or foci. Histologically, mononuclear cell infiltrates were seen to surround cholesterol crystals, in both the endometrium and the smooth muscle layer, reaching the serosa. The findings support the role of haemorrhage in promoting chronic inflammatory reactions around interstitial cholesterol ester precipitates., (Copyright 1999 Harcourt Publishers Ltd.)

Mixed mullerian tumor (MMT) is a biphasic malignancy of elderly women. It, including both a carcinomatous and a sarcomatous component (CC and SC), is regarded as a female genital tract carcinosarcoma (FGTCS). Since current methods to grade CC and SC are not still univocal, the authors estimate mitotic index (MI) and MIB 1-immunolabeling index (MIB 1-LI) as common prognostic indices for the MMT components. They also compare above prognostic indices with p-53 immunocytochemistry, in MMTs. The present study thus points out that: (a) MI of CC and SC areas is consistent with the respective conventional tumor grades; (b) MI averages of CC are higher than those observed in the SC areas; (c) MI and MIB 1-LI of the CC-tumor cells correlate reciprocally in a very significant fashion; (d) A diffuse strong p53 nuclear immunostaining (> 50% cells) is often patent where the highest MI and MIB 1-LI are found. In conclusion, the authors propose MI and MIB 1-LI as two complementary useful indices to assess prognosis of MMTs. They also suggest p53 nuclear immunolabeling should be regarded as an independent biomarker of unfavourable MMT behaviour.

Histologic and Transforming Growth Factor Beta (TGF-beta) immunostain patterns were sought in resected distal urinary tracts from 17 Primary Megaureter (PM) affected children, referred to surgery. Comparative observations were also carried out on embryonal and fetal ureteral buds of both humans and bovines. A reciprocal resemblance was mainly objectivized between the resected "narrowed" ureters of patients under 18 months, and the fetal ureteral buds at 26th and 38th gestational week. A development delay was irrespectively observed in PM "narrowed" ureters, at the longitudinal muscle-bundles in the parietal juxta-luminal compartment. A consistent TGF-beta immunostain cytoplasmic reaction there selectively depicted the growing mesenchymal lines, including both the undifferentiated single cells and the muscle-like profiled ones. These results agree with very recent reports perspecting a segmental maturation delay as a pathogenetic moment of PM. Because of the acquired potent TGF-beta inhibitory role on myoblasts differentiation, the present study substantiates a persistent TGF-beta role in perinatal ureter dilations.

Previous reports showed Helicobacter pylori (H. pylori) in type B gastritis-affected stomachs. This study was carried out to compare H. pylori staining effectiveness on biopsy to brush cytology. Tissue and brush parallel samples of gastric mucosa with abnormal or normal appearances were examined: 57.6% H. pylori-positive pieces from the antrum and 19.2% from the body were found, versus 65.3% and 25% H. pylori-positive brush smears, respectively. H. pylori resembling organisms were mainly related to chronic and acute antral inflammations and were often associated with higher amounts of round-shaped cocco-bacteria. In addition, H. pylori direct stain on brushing is proposed as the most rapid and reliable method for the routine diagnosis of Helicobacter pylori infection, in both ulcer or nonulcer gastritis.