Effects of ischaemic post-conditioning on the early and late testicular damage after experimental testis ischaemia-reperfusion.

Ischaemic post-conditioning (IPostC) might represent an innovative surgical approach to protect organs from ischaemia and reperfusion (I/R) injury. We investigated the molecular mechanisms underlying the contrasting effects of IPostC on the early and late damage induced by testicular I/R injury. Testis I/R was induced by occluding the right testicular vessels using a clip. Male rats were divided into the following groups: sham, I/R and I/R + IPostC. In the I/R group, the clip was removed after 60 min of ischaemia, and reperfusion was allowed for 30 min, 1 and 30 days. In the I/R + IPostC group, three cycles of 30-sec reperfusion and 30-sec ischaemia were performed after 60 min of ischaemia and then reperfusion followed up for 30 min, 1 and 30 days. Following 30-min reperfusion, there was an increase in mitogen-activated protein kinases (MAPKs) in I/R rats; after 1 day of reperfusion, interleukin-6, tumour necrosis factor-α and nuclear factor-κB (NF-κB) expression were significantly increased; IκB-α expression reduced; and a marked damage in both testes was observed. IPostC inhibited MAPKs, cytokines and NF-κB expression, augmented IκB-α expression and decreased histological damage in testes subjected to I/R. After 30 days of reperfusion, I/R injury activated the apoptosis machinery, caused severe histological damage and reduced spermatogenic activity. By contrast, IPostC did not modify the apoptotic markers, the histological alterations as well as spermatogenic activity following 30 days of reperfusion. Our data demonstrate that IPostC protects the testis from the early damage induced by I/R injury, but it does not protect against the late damage.

Effects of polydeoxyribonucleotide on the histological damage and the altered spermatogenesis induced by testicular ischaemia and reperfusion in rats.

The effects of polydeoxyribonucleotide (PDRN), an agonist of the A2A adenosine receptors which when activated positively influences sperm activity, were tested in an experimental testicular ischaemia/reperfusion injury model. Anaesthetized male Sprague-Dawley rats were subjected to testicular torsion-induced ischaemia, followed by reperfusion (TI/R). Immediately after detorsion, randomized animals, including SHAM, received intraperitoneal injections of: (i) vehicle (1 mL/kg 0.9% NaCl solution); (ii) PDRN (8 mg/kg); (iii) DMPX (3,7-dimethyl-1-propargilxanthine, 0.1 mg/kg); or (iv) PDRN (8 mg/kg) + DMPX (0.1 mg/kg). Animals were euthanized at 1, 7 and 30 days following reperfusion. Vascular endothelial growth factor (VEGF) expression is normally associated with adenosine A2A receptor stimulation. After treatment, VEGF mRNA/protein expression quantified by qPCR and Western blot, vascular endothelial growth factor receptor-1 (VEGFR1) and endothelial nitric oxide synthase (eNOS) mRNA measured by qPCR, VEGF and VEGFR1 assessed using immunohistochemical methods, histological staining and spermatogenic activity were all analysed. Testis ischaemia-reperfusion (TI/R) injury caused increases in VEGF mRNA and protein, VEGFR1 and eNOS mRNA, histological damage and reduced spermatogenic activity. Immunostaining showed a lower expression of VEGF in germinal epithelial cells and a strong expression of VEGFR1 in Leydig cells after TI/R. PDRN administration increased significantly VEGF message/protein, VEGFR1 and eNOS message, decreased histological damage and ameliorated spermatogenic activity. PDRN might be useful in the management of testicular torsion.

Aquaporin-9 immunohistochemistry in varicocele testes as a consequence of hypoxia in the sperm production site.

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.

Syndecan-1 and Wingless-type protein-1 in human ameloblastomas.

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).

Interstitial cells of Cajal network in primary obstructive megaureter.

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.

C-kit positive interstitial cells of Cajal network in primary obstructive megaureter.

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.

Endometrial polypoid adenomyomatosis in a bitch with ovarian granulosa cell tumour and pyometra.

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.

Small bowel intussusception by local recurrence of an inflammatory myofibroblastic tumor: report of a case and review of the literature.

Inflammatory myofibroblastic tumor (IMT) of the ileum is a rare, usually solitary lesion, that frequently presents small-intestinal intussusception and obstruction. We describe an IMT of the ileum in a 4.5-year old child who presented a small bowel intussusception. During laparotomy, an annular mass around the ileum was resected, and the IMT was histologically diagnosed. Three months after the operation, the patients were hospitalized with the symptoms of intestinal obstruction. Laparotomy showed a ileal intussusception. Along the previous suture line of anastomosis, a smooth polypoid tumor was evident. Segmental resection of the ileum, including the tumor mass, was performed. The IMT was immunohistochemically diagnosed. The patient was asymptomatic at 3 year follow-up. A review of the literature for this rare entity emphasizes the importance of immunohistochemical confirmation of its benign nature. Because of the risk of local recurrence, IMT cases should have a long-term follow-up.

Postmenopausal bleeding and vaginal nodules as the first presenting sign of adenocarcinoma of the gallbladder.

Vaginal submucosal nodules were observed in a 67-year-old woman, with ultrasonographic features of an advanced uterine neoplasm. On biopsy, light microscopy suggested that the lesions might be metastatic foci from an extragenital cancer, with a prevalent tubular growth pattern. Parallel immunohistochemical reactions revealed a diffuse, strong CA 19-9 positivity in both the cell membrane and cytoplasm. Subsequently, high serum levels of such tumor marker were also found, and an extragenital cancer was suspected of pancreatic or biliary origin. A mass in the gallbladder fossa was then detected by computed tomography and a primary gallbladder adenocarcinoma was confirmed on ultrasound-guided biopsy.

Hysteroscopic findings of endometrial carcinoma. Evaluation of 104 cases.

PURPOSE OF INVESTIGATION: Retrospective evaluation of hysteroscopic findings in the accurate diagnosis of endometrial carcinoma. METHODS: A retrospective monocentric study from January 1995 to December 2004. One hundred and four patients with hysteroscopic aspects evocative of endometrial carcinoma confirmed by endometrial biopsy during diagnostic hysteroscopy, by surgical hysteroscopic resection pieces or by hysterectomy specimen were included. RESULTS: Among the 104 patients, diagnostic hysteroscopy pointed out endometrial features suggestive of endometrial carcinoma in 102 cases. In two women diagnostic hysteroscopy failed to diagnose endometrial malignancy which was identified on pieces of polyps by surgical hysteroscopic resection. DISCUSSION: Polypoid proliferations cerebroid in appearance, with ulceration and necrosis, friable and with irregular vessels, represent endometrial findings highly indicative of malignancy. The diagnosis may be missed in cases of focal neoplasias, within endometrial polyps or in conditions of unsatisfactory endouterine visualization.

Immunoexpression of aquaporin-1 in adolescent varicocele testes: possible significance for fluid reabsorption.

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 case of bilateral prenatal testicular torsion: Ultrasonographic features, histopathological findings and management.

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.

Malignant mixed sex cord-stromal tumour in a stallion.

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.

Primary ovarian leiomyosarcoma. Proliferation rate and survival.

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.

Uterine malignant mixed Müllerian tumor (Metaplasic carcinoma) in the cat: clinicopathologic features and proliferation indices.

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.

[Do histologic changes of the upper renal pole in double ectopic ureterocele justify a conservative approach?]. / Le alterazioni istologiche del polo renale superiore nell'ureterocele ectopico duplex giustificano un approccio conservativo?

The aim of this study was to review the histology of the upper pole segment in patients with duplex ectopic ureterocele to verify if a less aggressive surgery is justified in the prenatally diagnosed patients. We reviewed the histology of the upper pole segment of 15 consecutive patients with duplex system ectopic ureterocele treated between 1991 and 1999 at the Paediatric Surgery Unit of University Hospital of Messina. The diagnosis of duplex system ectopic ureterocele was made according to the criteria of the Section on Urology of the American Academy of Paediatrics. The histology specimens were assessed for dysplastic, inflammatory and obstructive changes. All 15 patients with duplex system ectopic ureterocele were surgically treated with heminephro-ureterectomy and the surgical specimens were histologically examined. Nine of the 15 patients were prenatally diagnosed. The histology of the upper pole segment of the 9 prenatally diagnosed showed in all patients segmental renal microcystic dysplasia, chondroid metaplasic islands and an inflammatory tubulo-interstitial nephropathy in 6 patients (66.6%) and in 2 (22.2%) nephroblastomatosis. The histology of six the postnatal postnatally diagnosed patients showed in all patients segmental multicystic renal dysplasia, inflammatory tubulo-interstitial nephropathy and segmental parenchymal scars. The upper pole histology of the patients with duplex ectopic ureterocele diagnosed prenatally did not show any evidence of reversible histological change. Considering the histology and the good outcome of patients treated with upper pole nephroureterectomy a less aggressive surgery with preservation of the upper pole does not seem justified.