Microarrays of bladder cancer tissue are highly representative of proliferation index and histological grade.

The number of genes suggested to play a role in cancer biology is rapidly increasing. To be able to test a large number of molecular parameters in sufficiently large series of primary tumours, a tissue microarray (TMA) approach has been developed where samples from up to 1000 tumours can be simultaneously analysed on one glass slide. Because of the small size of the individual arrayed tissue samples (diameter 0.6 mm), the question arises of whether these specimens are representative of their donor tumours. To investigate how representative are the results obtained on TMAs, a set of 2317 bladder tumours that had been previously analysed for histological grade and Ki67 labelling index (LI) was used to construct four replica TMAs from different areas of each tumour. Clinical follow-up information was available from 1092 patients. The histological grade and the Ki67 LI were determined for every arrayed tumour sample (4x2317 analyses each). Despite discrepancies in individual cases, the grade and Ki67 information obtained on minute arrayed samples were highly similar to the data obtained on large sections (p<0.0001). Most importantly, every individual association between grade or Ki67 LI and tumour stage or prognosis (recurrence, progression, tumour-specific survival) that was observed in large section analysis could be fully reproduced on all four replica TMAs. These results show that intra-tumour heterogeneity does not significantly affect the ability to detect clinico-pathological correlations on TMAs, probably because of the large number of tumours that can be included in TMA studies. TMAs are a powerful tool for rapid identification of the biological or clinical significance of molecular alterations in bladder cancer and other tumour types.

High-throughput tissue microarray analysis of 3p25 (RAF1) and 8p12 (FGFR1) copy number alterations in urinary bladder cancer.

Studies by comparative genomic hybridization revealed that the chromosomal regions 3p25 and 8p11-p12 are recurrently amplified in bladder cancer. To investigate the prevalence of DNA copy number alterations in these chromosomal regions and study their clinical significance, we used probes for the RAF1 (3p25) and FGFR1 (8p12) genes for fluorescence in situ hybridization. A tissue microarray containing 2317 tumors was analyzed. The analysis revealed RAF1 amplification in 4.0% and FGFR1 amplification in 3.4% of interpretable tumors. In addition, deletions were found at the 3p25 locus in 2.2% and at the 8p11-12 locus in 9.9% of interpretable tumors. Both amplifications and deletions of RAF1 and FGFR1 were significantly associated with high tumor grade (P < 0.0001), advanced stage (P < 0.0001), and poor survival (P < 0.05) if tumors of all of the stages where analyzed together. RAF1 amplifications were associated with subsequent tumor progression in pT1 carcinomas (P < 0.05). The marked differences in the frequency of all of the analyzed changes between pTa grade 1/grade 2 and pT1-4 carcinomas support the concept of these tumor groups representing different tumor entities.

Choroideremia, sensorineural deafness, and primary ovarian failure in a woman with a balanced X-4 translocation.

We present clinical and cytogenetic studies of a female patient affected with choroideremia, mild sensorineural deafness, and primary amenorrhea showing a balanced translocation between chromosomes X and 4. The breakpoint was precisely defined applying FISH techniques: 46,X,t(X;4)(q21.2;p16.3).ish t(X;4)(D4S96+, D4F26+; wcpX+). The X-chromosomal breakpoint was located within a region where both the choroideremia locus and a deafness locus (DFN3/POU3F4) have been mapped. The presence of X-linked disorders in this balanced carrier of X-autosomal translocations (XAT) can be explained either by the disruption of the structural coding or regulatory sequences of the gene(s) or by the submicroscopic deletion of this region leading to a contiguous gene deletion syndrome. The primary ovarian failure (POF) found in the present case has been already observed in XAT when the breakpoint is within a previously defined critical region (Xq13-26). A position effect is postulated as a possible explanation.

High-throughput tissue microarray analysis of cyclin E gene amplification and overexpression in urinary bladder cancer.

Studies by comparative genomic hybridization revealed that the 19q13 chromosomal region is frequently amplified in bladder cancer. The cyclin E gene (CCNE), coding for a regulatory subunit of cyclin-dependent kinase 2, has been mapped to 19q13. To investigate the role of cyclin E alterations in bladder cancer, a tissue microarray of 2,317 specimens from 1,842 bladder cancer patients was constructed and analyzed for CCNE amplification by fluorescence in situ hybridization and for cyclin-E protein overexpression by immunohistochemistry. Fluorescence in situ hybridization analysis showed amplification in only 30 of the 1,561 evaluable tumors (1.9%). Amplification was significantly associated with stage and grade (P: < 0.0005 each). Immunohistochemically detectable cyclin E expression was strong in 233 (12.4%), weak in 354 (18.9%), and negative in 1, 286 of the 1,873 interpretable tumors. The majority (62.1%) of CCNE-amplified tumors were strongly immunohistochemistry-positive (P: < 0.0001). The frequency of protein expression increased from stage pTa (22.2%) to pT1 (45.5%; P: < 0.0001) but then decreased for stage pT2-4 (29.4%; P: < 0.0001 for pT1 versus pT2-4). Low cyclin E expression was associated with poor overall survival in all patients (P: < 0.0001), but had no prognostic impact independent of stage. It is concluded that cyclin E overexpression is characteristic to a subset of bladder carcinomas, especially at the stage of early invasion. This analysis of the prognostic impact of CCNE gene amplification and protein expression in >1,500 arrayed bladder cancers was accomplished in a period of 2 weeks, illustrating how the tissue microarray technology remarkably facilitates the evaluation of the clinical relevance of molecular alterations in cancer.

Rapid identification of a small dicentric supernumerary marker derived from chromosome 16 with a modified FISH technique on amniotic fluid.

Small supernumerary marker chromosomes are seldom found in prenatal diagnosis and the majority of them are difficult to identify. The only possibility to give a more precise prognosis is by establishing its origin. FISH is the best technique to identify the chromosomal origin, but in the majority of cases large amounts of chromosomal material are needed and this is time consuming. We have used a modification of the FISH technique that allows the hybridization of several probes on one slide. Using this method, we have identified the first de novo mosaic dicentric supernumerary marker derived from chromosome 16 (smaller than chromosome 21) in amniotic fluid. The gestation and the follow-up of the baby were normal.

Lymphocytic colitis: clinical presentation and long term course.

BACKGROUND: Lymphocytic colitis is characterised by chronic watery diarrhoea with normal endoscopic or radiological findings and microscopic evidence of pronounced infiltration of the colonic mucosa with lymphocytes. AIM: To investigate the long term clinical and histological evolution of the disease in a large group of patients with well characterised lymphocytic colitis. METHODS: Between 1986 and 1995 the histological diagnosis of lymphocytic colitis was obtained in 35 patients; 27 of these agreed to a follow up examination. All clinical, endoscopic, and histopathological records were reviewed at that time and the patients had a second endoscopic examination with follow up biopsies. RESULTS: The patients initially presented with the typical findings of lymphocytic colitis. After a mean (SD) follow up of 37.8 (27.5) months, diarrhoea subsided in 25 (93%) and histological normalisation was observed in 22 (82%) of the 27 patients. Progression from lymphocytic colitis to collagenous colitis was not observed. CONCLUSIONS: Lymphocytic colitis is characterised by a benign course with resolution of diarrhoea and normalisation of histology in over 80% of patients within 38 months. Considering the benign course of the disease, the potential benefit of any drug treatment should be carefully weighed against its potential side effects.

Cutaneous lung tissue heterotopia.

AIMS: We describe the clinical and pathological findings of a previously unreported cutaneous lung tissue heterotopia in a child. CASE DETAILS: The 3-year-old female patient developed a 8-mm secreting papule over her left scapula. Pathological examination revealed a lesion composed of bronchioles and alveoli within the subcutis and the dermis, with bronchiolar connection to the epidermis. Alveolar type II cells indicating full pulmonary differentiation were detected with a monoclonal antibody (AMH 152). CONCLUSIONS: The described features suggest that this lesion is a unique variant of rarely observed bronchus-like entities of the skin, designated as cutaneous bronchogenic cysts or cutaneous branchial cleft cysts.

Transitional cell carcinoma of the vagina with pagetoid spread pattern.

A distinctive variant of a papillary noninvasive transitional cell carcinoma (TCC) of the vagina removed from a postmenopausal woman is described. The neoplasm was evaluated by immunohistochemistry. The designation of this neoplasm as a TCC is supported by its morphological features and its coexpression for cytokeratin (CK) 7 and CK 20. Its main feature is pagetoid infiltration into adjacent vaginal epithelium. This is the second reported case involving a transitional cell metaplasia (TCM) of the vagina, a possible precursor lesion of the TCC.

[Cholesterol crystal embolisms due to systemic thrombolysis of an acute myocardial infarct]. / Cholesterinkristall-Embolien infolge systemischer Thrombolyse eines akuten Myokardinfarkts.

A 63-year-old male patient admitted with acute inferior myocardial infarction was treated with tissue plasminogen activator (rt-PA) and heparin. 4 hours after the initiation of the rt-PA infusion he showed painful cutaneous alterations on the lower trunk, mistaken as drug-induced rash and later correctly identified as livedo reticularis. Simultaneously, renal function deteriorated. Assuming hypersensitivity vasculitis, we instituted immunosuppressive treatment which proved to be ineffective. Skin biopsy including the deeper layers showed multiple cholesterol emboli. Anticoagulants were stopped and we noticed no further cholesterol embolism or further decline of renal function. 10 previous publications have mentioned a causal relationship between systemic fibrinolytic treatment and the cholesterol crystal embolism syndrome. In 7 cases, however, angiographic procedures were used or the interval between fibrinolysis and the occurrence of cholesterol crystal embolism was too long to exclude spontaneous or heparin-induced cholesterol crystal embolism. The short interval in our own case points clearly to the systemic fibrinolytic therapy as the culprit. We would like to draw attention to the possibility that in individuals with advanced atherosclerotic lesions of the aorta or major arteries, systemic fibrinolytic treatment of myocardial infarction may give rise to the cholesterol crystal embolism syndrome.

[Microangiopathic hemolytic anemia, erythrophagocytosis and consumption coagulopathy in vinyl chloride-induced hemangiosarcoma of the spleen and liver]. / Mikroangiopathische hämolytische Anämie, Erythrophagozytose und Verbrauchskoagulopathie bei vinylchloridinduziertem Hämangiosarkom von Milz und Leber.

A case of angiosarcoma of the liver and the spleen following vinyl chloride exposure is described. The main symptoms in clinical diagnosis were microangiopathic hemolysis, disseminated intravascular coagulation, hepatosplenomegaly and exposure to vinyl chloride thirty years ago. It is the first case in which liver and spleen are involved in angiosarcoma due to vinyl chloride exposure. The tumor cells showed angioformative and solid histiocytoid growth with erythrophagocytosis.

[Collagenous sprue and ulcerative jejuno-ileitis in a patient with gluten-induced enteropathy]. / Kollagensprue und ulzerierende Jejunoileitis bei einem Patienten mit gluteninduzierter Enteropathie.

In a 60-year-old patient with adult celiac disease complete clinical remission was achieved with a glutenfree diet, but symptoms of sprue subsequently recurred despite rigid adherence to dietary restrictions. Further investigations revealed simultaneous occurrence of two severe complications of celiac disease: collagenous sprue and ulcerative jejunoileitis. The patient died after a 2 1/2 year course of refractory, severe, relentlessly progressive malabsorption. In the light of the literature and this observation it is postulated that in every case of primary or secondary refractory celiac disease complications must be sought: to diagnose collagenous sprue, aspiration biopsy from the proximal jejunum, and possibly also from the terminal ileum, may be more efficient than simple endoscopic biopsy from the descending duodenum. If the diagnosis of collagenous sprue can be established, therapy with steroids should be attempted. The diagnosis of ulcerative jejunoileitis is usually missed by radiology and requires exploratory laparotomy.

[Cellular immune deficiency and polylymphadenopathy in the male sexual partners of a woman who died of acquired immunodeficiency (AIDS)]. / Zellulärer Immundefekt und Polylymphadenopathie bei männlichen Sexualpartnern einer an erworbenem Immundefekt (AIDS) verstorbenen Frau.

A report is presented on a woman who died of acquired immune deficiency syndrome (AIDS) and her two sexual partners. One of the men had generalised lymphadenopathy and the other severe depression of helper-T-cells with cutaneous anergy, symptoms which may be observed in the prodromal state of AIDS. Apart from regular sexual contact with the woman, the two men had no further risk factors for AIDS.

[Collagen colitis. A new disease picture or merely a histological abnormality?]. / Die "Kollagenkolitis". Ein neues Krankheitsbild oder blosse histologische Abnormität?

"Collagenous colitis" has been recently defined as a new entity characterized by protracted diarrhoea, associated with subepithelial broadening of the collagen layer. A report is presented on two patients fulfilling the above criteria. Based on a critical analysis of these two cases and on prospective study of biopsies in 50 consecutive coloscopies it is concluded that a causal relationship between the enlarged collagen layer and the diarrhoea cannot be unequivocally established and hence "collagenous colitis" should not yet be considered a new entity.

[Immature solid teratoma of the uterine cervix (author's transl)]. / Unreifes solides Teratom der Portio uteri.

A 27-year-old woman with an inconspicuous anamnesis developed within two years a solid tumor of about 10/5 cm in size, and extending from the left corner of the external os up to the lumen of the vagina and consisting of partly immature ectodermic, mesodermic and endodermic parts. After the radical removal of the tumor by means of abdominal hysterectomy and left-sided adnexectomy no signs of a recurrence could be observed for about ten years. Except a shortly described case of teratoma of the endometrium no further case of a uterine teratoma has been mentioned in the literature so far. A brief discussion of differential diagnosis and etiology is also present.

Effects of quinolinic acid and glucagon on gluconeogenesis in the perfused rat liver.

Previous findings that 2.5 mM quinolinic acid inhibits gluconeogenesis more strongly from alanine than from lactate have been confirmed. 15 mM quinolinic acid completely inhibited gluconeogenesis from lactate as well as from alanine whereas the formation of glucose from fructose and the production of urea from ammonia and lactose were not affected. The pattern of the gluconeogenic intermediates was the same in the presence of 15 mM quinolinic acid as with 2.5 mM of the inhibitor. It is concluded that high as well as low concentrations of quinolinic acid inhibit gluconeogenesis at the step between oxaloacetate and phosphoenolpyruvate. Furthermore, 5-methoxyindole-2-carboxylic acid, an inhibitor of mitochondrial pyruvate metabolism, also completely blocked gluconeogenesis from lactate whereas glycerol conversion to glucose was only weakly inhibited. All these results do not support the concept of an alternate pathway of gluconeogenesis from lactate proposed by others. 2.5 mM quinolinic acid also partially blocked the formation of urea from alanine. It is suggested that quinolinic acid may have a second site of action causing an inhibition of the glutamate-pyruvate transamination owing to lack of 2-oxoglutarate in the cytosol. In the presence of quinolinic acid, glucagon caused about the same increase in aspartate and malate tissue levels in the absence of added substrates as in the presence of added lactate or alanine. Therefore, no additional effect of glucagon on gluconeogenesis from lactate or alanine prior to the block by quinolinic acid could be demonstrated.