Changes in the expression profiles of claudins during gonocyte differentiation and in seminomas.

Testicular germ cell tumors (TGCTs) are the most common type of cancer in young men and their incidence has been steadily increasing for the past decades. TGCTs and their precursor carcinoma in situ (CIS) are thought to arise from the deficient differentiation of gonocytes, precursors of spermatogonial stem cells. However, the mechanisms relating failed gonocyte differentiation to CIS formation remain unknown. The goal of this study was to uncover genes regulated during gonocyte development that would show abnormal patterns of expression in testicular tumors, as prospective links between failed gonocyte development and TGCT. To identify common gene and protein signatures between gonocytes and seminomas, we first performed gene expression analyses of transitional rat gonocytes, spermatogonia, human normal testicular, and TGCT specimens. Gene expression arrays, pathway analysis, and quantitative real-time PCR analysis identified cell adhesion molecules as a functional gene category including genes downregulated during gonocyte differentiation and highly expressed in seminomas. In particular, the mRNA and protein expressions of claudins 6 and 7 were found to decrease during gonocyte transition to spermatogonia, and to be abnormally elevated in seminomas. The dynamic changes in these genes suggest that they may play important physiological roles during gonocyte development. Moreover, our findings support the idea that TGCTs arise from a disruption of gonocyte differentiation, and position claudins as interesting genes to further study in relation to testicular cancer.

[Iris abscess after bacterial endocarditis in a patient with leukaemia. Differential diagnosis]. / Absceso iridiano tras endocarditis bacteriana en un paciente con leucemia. Diagnóstico diferencial.

PURPOSE: To report a case of iris abscess due to bacterial endocarditis. CASE REPORT: A 46-year-old male under diagnosis of promielocitic leukemia and endocarditis presented with decreased vision in left eye (OS). Ophthalmic exploration revealed iris abscess and hypopyon with fibrinous exudate in iris of the left eye and tyndall +1 in right eye (OD). Blood culture and anterior chamber paracentesis was positive for methicillin-sensitive Staphylococcus aureus and negative for blastic cells in citology. Treatment with systemic antibiotic was initiated with total resolution of inflammation. CONCLUSION: Iris abscess is an unusual septic focus in bacterial endocarditis. It is crucial to rule out an extramedullary metastasis in a patient with leukemia due to the general prognosis.

The neuroendocrine component in bladder tumors.

Neoplastic urothelium has the capacity to display enormous plasticity and divergent differentiation. Neuroendocrine tumors arise as a result of such capacity. Neuroendocrine tumors of the bladder represent a limited number of neoplasms characterized by neuroendocrine hormone secretion and a poor outcome. These tumors can be displayed as pure neuroendocrine neoplasms or more frequently as a neuroendocrine counterpart mixed with classical urothelial bladder cell carcinomas, adenocarcinoma, sarcomatoid carcinoma or mixtures of these components. Their heterogeneous character and clinical aggressiveness remain a challenge for clinical, pathological diagnosis and for therapy selection. Several types have been described, although small cell carcinoma represents the major subgroup of neuroendocrine tumors as compared to large cell carcinoma and carcinoid subtypes. In this review, epidemiology, presentation, macroscopic and microscopic features, and clinical prognostic and therapeutic implications of the major subgroups are described. Special focus is given to discuss how immunohistochemistry protein patterns and laboratory determinations may aid to characterize this type of tumors and to improve the clinical management of this highly aggressive type of bladder cancer patients.

Fine needle aspiration cytology of basal cell adenoma of the salivary gland: a cytohistological correlation study of 35 cases.

OBJECTIVE: In order to evaluate the possibility of a specific cytological recognition of basal cell adenoma (BCA) we reviewed our experience with 35 histologically proven cases. Few series describing cytological features of BCA are available and diagnostic cytological criteria are not well established. METHODS: This study was based on 41 cytology samples from 35 patients with BCA. Thirty-five aspiration procedures were performed pre-operatively and six on tumour recurrence. Nineteen of the 35 patients were men and 16 women. The mean age at diagnosis was 55 years old (range 24-92). The series includes one non-representative case. Except for one tumour located in the upper lip, all of them involved the parotid gland. RESULTS: Aspirates were cellular, showing groups with dense, homogeneous metachromatic stroma and single cells. Relevant features were the trident-like configuration of groups, intimate relationship between neoplastic cells and stroma and cellular polymorphism. In approximately half of the cases a precise diagnosis was given. Most of the remaining tumours were diagnosed as benign but they were difficult to differentiate from pleomorphic adenoma. Regarding malignancy, there were two misdiagnoses of acinic cell carcinoma, due to high epithelial cellularity along with scarcity of stroma, and one case was considered to be suspicious of malignancy. CONCLUSION: BCA shows characteristic cytological features that allow a precise diagnosis. The main differential diagnosis is epithelial-rich pleomorphic adenoma, while acinic cell carcinoma is a potential false positive.

Role of fine needle aspiration cytology in the diagnosis and management of Warthin's tumour of the salivary glands.

OBJECTIVE: Local excision surgical procedures and non-surgical conservative management are considered alternatives to superficial parotidectomy in the treatment and management of Warthin's tumour (WT). Such therapeutic alternatives demand accurate diagnosis. In order to determine whether fine needle aspiration cytology (FNAC) is capable of rendering such a minimally invasive diagnosis, we evaluated its accuracy and diagnostic parameters in a large series of histologically proven cases of WT. METHODS: A cytohistological study of 116 salivary tumours from 110 patients (four WT were bilateral) with a histological or cytological diagnosis of WT. RESULTS: Histology confirmed the cytological diagnosis in 103 of 114 tumours (90.4%). Two tumours were incorrectly diagnosed on cytology as WT. In 11 cases of WT there was an erroneous or non-representative cytological diagnosis. The sensitivity was 90.4%, and positive predictive value 98.1%. Regarding malignancy, there were three misdiagnoses. One tumour diagnosed as WT was a low-grade mucoepidermoid carcinoma. Two cases considered 'suspicious of squamous cell carcinoma' corresponded to WT. After review, 81.3% of the cases of WT were considered typical and 18.7% non-typical; all misdiagnoses were in the latter group. Cytological difficulties could be divided into three areas: (i) absence of one or more diagnostic components; (ii) 'squamoid' pattern; and (iii) mucinous metaplasia. Degenerated oncocytes were present in 65% of cases. CONCLUSIONS: FNAC offers the possibility of a reliable diagnosis of WT. Pathologists must pay attention to the squamous appearance of degenerated oncocytes. Cytology, when coupled with clinical and image findings, may permit conservative tumour management.

Cytological features of chromophobe renal cell carcinoma, classic type. A report of nine cases.

OBJECTIVE: To review our experience with nine cases of chromophobe renal cell carcinoma (ChRCC), classic type. The cytological descriptions of this entity are still rare, and information concerning the diagnostic value of cytology is needed. METHODS: Nine cases of ChRCC evaluated using fine needle aspiration (n = 6) or intraoperative scrape cytology (n = 3) were selected. Expression of vimentin was evaluated in four cases using immunocytochemistry, which was performed on alcohol-fixed material. In all cases a complete pathological study was available. RESULTS: The neoplastic cells were arranged mainly as single cells and small, discohesive, monolayered groups. A polymorphous cellular population was identified, with coexisting large, small and intermediate-sized cells. The large neoplastic cells showed clear, flocculent cytoplasm with small, eccentric nuclei and frequent binucleation. Dense, homogeneous cytoplasm was most commonly seen in smaller cells. Clear cytoplasmic spaces resembling perinuclear halos were frequently observed, best appreciated in cells with more dense cytoplasm. Binucleation and a marginal nuclear location were commonly seen. Necrosis, basement membrane or other stromal material were absent. Vimentin was not expressed in the four cases analysed. Precise cytological recognition was possible in the last five cases. CONCLUSIONS: There is increasing evidence that a cytological diagnosis of ChRCC is possible. In our experience the histopathological features of ChRCC were well reflected in cytological samples, allowing specific recognition. In our cases the main differential diagnosis considered was clear cell carcinoma. Cytology can be especially helpful in the evaluation of intraoperative samples.

Discovery of myopodin methylation in bladder cancer.

Myopodin is an actin-binding protein that shuttles between the nucleus and the cytoplasm. After identifying an enriched CpG island encompassing the transcription site of myopodin, we aimed at evaluating the potential relevance of myopodin methylation in bladder cancer. The epigenetic silencing of myopodin by hypermethylation was tested in bladder cancer cells (n=12) before and after azacytidine treatment. Myopodin hypermethylation was associated with gene expression, being increased in vitro by this demethylating agent. The methylation status of myopodin promoter was then evaluated by methylation-specific polymerase chain reaction (MS-PCR) analyses. Myopodin was revealed to be frequently methylated in a large series of 466 bladder tumours (68.7%). Myopodin methylation was significantly associated with tumour stage (p<0.0005) and tumour grade (p=0.037). Myopodin expression patterns were analysed by immunohistochemistry on tissue arrays containing bladder tumours for which myopodin methylation was assessed (n=177). The presence of low nuclear myopodin expression alone (p = 0.031) or combined with myopodin methylation (p=0.008) was associated with poor survival. Moreover, myopodin methylation in 164 urinary specimens distinguished patients with bladder cancer from controls with a sensitivity of 65.0%, a specificity of 79.8%, and a global accuracy of 75.3%. Thus, myopodin was identified to be epigenetically modified in bladder cancer. The association of myopodin methylation and nuclear expression patterns with cancer progression and clinical outcome, together with its ability to detect bladder cancer patients using urinary specimens, suggests the utility of incorporating myopodin methylation assessment in the clinical management of patients affected by uroepithelial neoplasias.

[Usefulness of cytology in diagnosis and management of renal and perirenal tumors in adults]. / Utilidad de la citología en el diagnóstico y manejo de tumores renales y perirrenales del adulto.

INTRODUCTION: The use more and more extended of tumorectomy, partial nephrectomy and nonsurgical treatments of renal tumors has supposed a renewed interest in the diagnosis use of cytology. Whether during preoperative period, through the puncture aspiration with fine needle (PAAF), or during the intraoperative analysis, the cytology offers the possibility of a specific morphologic diagnosis. In this revision the information concerning the diagnostic value of the cytology in renal tumors is updated. MATERIAL AND METHODS: The references related to renal masses cytological descriptions has been reviewed. For this purpose we have searched both with computer in Medline data base and also manually. In the same way we include authors experience as much in the PAAF of these lesions as in the intraoperative use of the cytology. RESULTS: Between neoplasias with more cytological typical presentation are the clear cell renal and papillary carcinomas. The chromophobe and oncocytoma can show similarities, although the accumulated experience in the last years reflects that its differentiation is possible in most of the cases. For the diagnosis of angiomyolipoma, urothelial carcinoma and kidney metastasis, the clinical and image information are of great interest for the pathologist. The integration of these data usually allows a specific diagnosis. CONCLUSION: Generally, cytology reflects with accuracy the histological characteristics of renal neoplasias, allowing in many cases a specific diagnosis. We consider much appropriated the use of cytology, due to the more and more frequent situation of "incidentaloma". The PAAF minimum invasive nature and the possibility of performing a fast cytological analysis during intraoperative studies offer important information for the therapeutic management of these patients.

[Histological variants of urothelial carcinoma with diagnostic, prognostic and therapeutical implications]. / Variantes histológicas del carcinoma urotelial con implicaciones diagnósticas, pronósticas y terapéuticas.

Most of urothelial carcinomas (UC) have a pattern of histological growth of papillary or solid type; nevertheless, in some cases there are histological types that significantly differ from these habitual patterns In this paper we have selected those UC variants that by its diagnosis difficulty and therapeutical or prognosis implications have to be perfectly identified and known by pathologists and urologist. The variants that we have considered of greater clinical and pathological interest have been: tubular and/or nested UC, microcystic UC, micropapillary UC, lymphoepithelioma like UC, plasmacytoid UC and sarcomatoid UC. A revision of the literature has been made of each one of these patterns evaluating the criteria diagnoses, clinical behavior and the present therapeutic options. In addition, we suggest that these UC variants must be explicitly reflected in the pathology report, due to its clinical implications.

[Diagnostic value of the gonadal biopsy in the disorders of sex development]. / Valor de la biopsia gonadal en el diagnóstico de los desórdenes del desarrollo sexual.

The gonadal biopsy provides essential information for the identification, classification and early detection of neoplasias in patients with disorders of sex development. Histopathological findings in these cases must be analysed together with clinical, hormonal, genetic and molecular information before deciding a therapeutic option. Sexual differentiation is the result of multiple and complex genetic and endocrinal mechanisms; therefore, we first present the events taking place during gonadal embryonic development, focusing on the genetic mechanisms involved in sexual determination and the differentiation of the testis and the urogenital tract. In second place, we describe the different gonads in the intersexual states -in testicular regression syndrome, fibrous streak, testicular dysgenesis, streak testes, ovotestes and microscopically normal testes and ovaries-, highlighting the histological features and the differential findings that allow the pathologist to distinguish between these entities with the aid of clinical, genetic, hormonal and molecular information that are characteristic for each situation. In third place, we studied the incidence of neoplasias in gonadal dysgenesis, male pseudohermaphroditism and true hermaphroditism. Finally, we discuss the limitations of gonadal biopsy to achieve a correct diagnosis in the disorders of sex development.

Fine needle aspiration cytology of mammary carcinoma with osteoclast-like giant cells.

Carcinoma with osteoclast-like giant cells (OCGC) is an uncommon neoplasm characterized by giant cells, prominent vascularization, haemorrhage and areas of cribriform epithelial growth with moderate atypia. Multinucleated giant cells (MGC) have been described in several other breast lesions raising an interesting differential diagnosis, mainly with benign disorders. Due to its rarity few cases have been described cytologically. We retrospectively reviewed 13 fine needle aspiration samples from nine patients with this variant of carcinoma. Nine corresponded to breast tumours and four to axillary, liver, subcutaneous and mediastinal metastatic lesions. The expression of CD68 by giant cells was evaluated immunocytochemically in six cases. All patients had a complete pathological study of the breast neoplasm. Smears showed a double component of epithelial and giant cells. Epithelial clusters were predominantly of intermediate size with irregular contours. Most were cohesive but others showed cellular dissociation with scarce to moderate cellular pleomorphism. Giant cells had well defined, deeply stained cytoplasm and round to elongated morphology. Two metastatic cases were devoid of them. Haemosiderin-laden macrophages were common in smears from breast tumours. In the six cases tested CD68 was expressed in MGC. Cytological features of mammary carcinoma with OCGC correlate closely with the histological ones. Most cases are clearly recognizable as malignant but in others cytological atypia may be minimal, mimicking a benign lesion. In difficult cases the presence of haemosiderin-laden macrophages and the histiocytic nature of the MGC are helpful diagnostic features.

Fine-needle aspiration cytology of Kikuchi's lymphadenitis: a report of ten cases.

The purpose of this study was to evaluate the cytologic features of Kikuchi's lymphadenitis (KL). Smears from 10 patients with histologically proven KL were reviewed. In all cases, fine-needle aspiration (FNA) was performed prior to biopsy. To assess the validity of morphologic recognition, a blinded study, including smears from non-Hodgkin's lymphomas, nonspecific, and mycobacterial lymphadenitis was performed. At least 5 cases showed characteristic cytologic findings that permitted their specific recognition. A polymorphous lymphoid population with abundant karyorrhectic debris and histiocytes, many of which showed a small size and eccentrically placed, crescent nuclei, were characteristic features of KL. The remaining 5 cases failed to show typical findings and were indistinguishable from other nonspecific, reactive lymphadenopathies. When typical cytologic findings are present in an adequate clinical context (cervical nodes in young patients), a precise diagnosis is possible, avoiding unnecessary biopsies.

Fine-needle aspiration cytology of "proximal-type" epithelioid sarcoma.

The cytologic and immunocytologic findings in a case of recurrent "proximal-type" epithelioid sarcoma (ES) of the vulva are presented. This is a recently described neoplasm that differs clinically and morphologically from conventional ES. Cytologic smears showed a dissociated population of large, atypical neoplastic cells with bi- and multinucleated cells, abundant cytoplasm, and rhabdoid-like morphology. Due to its different clinical management it must be differentiated from metastatic carcinoma and melanoma. From a practical perspective, its differentiation from other epithelial-like sarcomas is less important. In conclusion the cytopathologic findings of "proximal-type" ES show a good correlation with histopathology, permitting the diagnosis of recurrences and metastases. When accompanied by adequate clinical information and ancillary studies, a specific preoperative recognition seems possible.

Lipomembranous fat necrosis in three cases of testicular torsion.

AIMS: To describe for the first time a lesion termed lipomembranous fat necrosis (LFN) in three patients with spermatic cord torsion. METHODS AND RESULTS: We reviewed 386 testes and their epididymides and spermatic cords which had been removed for testicular infarction. For the three cases showing LFN, a battery of histochemical tests (including periodic acid-Schiff (PAS), orcein, Sudan black and Perls stains) was applied and clinical histories and laboratory data were also investigated. Findings were similar in the three specimens. The testes showed a central group of necrotic seminiferous tubules which were surrounded by granulation tissue consisting of macrophages, multinucleated giant cells, lymphocytes, plasma cells and fibrous connective tissue at the periphery of the lesion. The spermatic cord showed thrombosed veins surrounded by fat necrosis showing cystic cavities which were bounded by wavy hyaline membranes. These stained with Sudan black, PAS (before and after diastase digestion) and orcein and presented yellowish-green autofluorescence. CONCLUSIONS: Lipomembranous fat necrosis of the spermatic cord is a distinctive entity which seems to be related to spermatic cord torsion and the differential diagnosis of which should be established with regard to the presence of parasites, sclerosing lipogranuloma and granuloma evoked by rupture of a testicular prosthesis.

Epididymal growth and differentiation are altered in human cryptorchidism.

Despite the knowledge and histological classification of testicular lesions, epididymal lesions associated with cryptorchidism are not well defined and only macroscopic alterations have been reported. We have evaluated the alterations in the growth of both the epithelium and muscular wall of efferent ducts and epididymis in human patients with cryptorchidism from infancy to adulthood. In addition, by cytokeratin immunostaining we have also evaluated the stage of differentiation of each segment along the human postnatal life in these patients. A decrease is shown in the size of efferent and epididymal ducts in cryptorchid children compared with normal, age-matched controls. The height of the epithelium, muscular wall, and lumen of the cryptorchid epididymis were reduced at every age studied. This decrease in all regions was seen even in the testicular quiescent period (1 to 4 years of age). In addition, the cryptorchid epididymis grows more slowly during the transition to the pubertal period. The smaller size of the cryptorchid epididymis in pubertal and adult men compared with that of normal men is due primarily to underdevelopment of the muscular wall and a reduction in epithelial height. The pattern of growth of cryptorchid efferent ducts and ductus epididymides parallels that in normal men, except that development of the lumen and muscular layer in the cauda epididymis region are delayed. Epithelial differentiation, monitored by cytokeratin expression, is minimal in efferent ducts and throughout the epididymis of the cryptorchid male, and this is already seen in children. In conclusion, our immunohistochemical and morphometric results show a reduced development of the human cryptorchid epididymis that is already evident in childhood. They indicate that cryptorchidism is a primary congenital illness of the testis and spermatic ducts, with evident lesions from the first years of life, and suggest that surgical descent would probably not be able to completely reverse these alterations.

Androgen receptor expression in sertoli cells as a function of seminiferous tubule maturation in the human cryptorchid testis.

Androgen receptor (AR) immunohistochemistry was performed in an archival collection of adult human cryptorchid testes to determine whether AR cellular distribution and intensity of immunostaining were functions of the severity of cellular dysgenesis. The seminiferous tubule histology of cryptorchid testes collected from adults is marked by three specific patterns. 1) Seminiferous tubules are characterized as maintaining focal areas of germinal cell differentiation (albeit incomplete) that are interspersed with 2) tubules composed of Sertoli cells only, these latter cells being principally of the adult type, although dysgenetic and immature Sertoli cells may also be detected. 3) In contrast, there is a class of tubule that is characterized as being composed exclusively of Sertoli cells that are extremely dysgenetic in appearance. The majority of adult-type Sertoli cells found in the first types of tubules exhibited either robust or moderate AR staining intensity. Peritubular cells of these tubules also expressed a similar AR staining intensity. In contrast, in the more dysgenetic and immature type Sertoli cells found in the second type of tubules, the intensity of AR staining was significantly less, if not missing altogether. Finally, in the most dysgenetic tubules, Sertoli cell AR staining was never detected. To our knowledge, this is the first report in the literature that addresses the intensity of AR immunostaining in Sertoli cells of cryptorchid testes. The results presented herein are consistent with the interpretation that the intensity of AR staining in Sertoli cells diminishes as a function of the severity to which the cells are afflicted within a cryptorchid testis and that focal absence of AR expression in Sertoli cells correlates with a lack of local spermatogenesis in the tubules.