E2F3 is the main target gene of the 6p22 amplicon with high specificity for human bladder cancer.

Amplification of 6p22 occurs in about 10-20% of bladder cancers and is associated with enhanced tumour cell proliferation. Candidate target genes for the 6p22 amplicon include E2F3 and the adjacent gene NM_017774. To clarify which gene is representing the main target, we compared the prevalence of the amplification and the functional role of both genes. Amplification of E2F3 and NM_017774 was analysed by fluorescence in situ hybridization on a bladder cancer tissue microarray composed of 2317 cancer samples. Both genes showed amplification in 104 of 893 (11.6%) interpretable tumours and were exclusively found co-amplified. Additional gene expression analysis by real-time polymerase chain reaction in 12 tumour-derived cell lines revealed that amplification of 6p22 was always associated with co-overexpression of E2F3 and NM_017774. Furthermore, RNA interference was used to study the influence of reduced gene expression on cell growth. In tumour cells with and without the 6p22 amplicon, knockdown of E2F3 always lead to unequivocal reduction of proliferation, whereas knockdown of NM_017774 was only capable to slow down cell proliferation in non-amplified cells. Our findings point out that E2F3 but not NM_017774 is driving enhanced proliferation of 6p22 amplified tumour cells. We conclude that E2F3 must be responsible for the growth advantage of 6p22 amplified bladder cancer cells.

Role of complement and polymorphonuclear cells in demethylchlortetracycline-induced phototoxicity in guinea pigs. Inhibition by decomplementation in vivo.

In this study, demethylchlortetracycline was used as a prototype of exogenous phototoxic substances. In vitro, exposure of serum containing demethylchlortetracycline to ultraviolet-A irradiation resulted in the diminution of total complement hemolytic activity and C4, C2, C3, and C5 activities. In addition, chemotactic activity for human polymorphonuclear cells was generated, which was thermostable and antigenically related to human C5 but not human C3. In vivo, phototoxic lesions were induced in guinea pigs upon intradermal injections of demethylchlortetracycline solution, followed by ultraviolet-A irradiation. On a scale of 0-3+, the animals developed a maximal response of 2.5 at 20 h. This clinical response was associated with cellular infiltrate in the dermis, consisting of 29 +/- 2% of neutrophils at 24 h. The participation of the polymorphonuclear cells was evaluated in guinea pigs rendered neutropenic by treatment with cyclophosphamide. In these guinea pigs, demethylchlortetracycline and ultraviolet-A induced a maximal response of 0.75 +/- 0.5, which was associated histologically with 1.2 +/- 0.5% neutrophils in the dermis. The role of complement in this process was studied in guinea pigs congenitally deficient in C4, and in guinea pigs decomplemented by treatment with cobra venom factor. In contrast to normal guinea pigs, C4-deficient animals exhibited a maximal reaction of 0.83 +/- 0.16 at 6 h, which subsided within 24 h. Cobra venom factor-treated guinea pigs developed a maximal response of 0.5 at 0.5 and at 6 h. These clinical changes were associated with the development of an increased vascular permeability, as demonstrated by studies using guinea pigs injected intravenously with Evans blue solution. In animals with a normal complement system, there was intense localized bluing at the sites of phototoxic lesion. In contrast, only minimal bluing was observed in decomplemented guinea pigs. These data indicate that a normal number of polymorphonuclear cells and an intact complement system are required for the full development of demethylchlortetracycline-induced phototoxic lesions.

Infrared thermography in the detection and management of coronary artery disease.

Infrared thermography was used to measure and map precordial skin temperature in 60 patients undergoing elective coronary angiography; 9 patients were normal and 51 had coronary artery disease (CAD). Thermograms were graded by quartile area (zero to 4 plus) and magnitude of thermal asymmetry (recorded as degrees celsius). The presence, mean area and degree of thermal asymmetry were significantly greater in patients with CAD. Twenty-two patients subsequently underwent successful revascularization with angioplasty with a highly significant decrease in the presence, magnitude and degree of thermal asymmetry. The results demonstrate that CAD is associated with precordial thermal asymmetry. The area and magnitude of thermal asymmetry is greater in patients with CAD than in control subjects without angiographically significant CAD. Successful revascularization changed the asymmetric precordial pattern to a more symmetric one. Infrared thermography is a promising technique for the detection of CAD before and after revascularization.

[Evaluation of potential target genes of the 6p22.3-amplicon in urinary bladder cancer]. / Evaluation von potentiellen Zielgenen innerhalb des 6p22.3-Amplikons beim Urothelkarzinom der Harnblase.

Amplification of 6 p22.3 is one of the most frequent chromosomal alterations in high grade and invasive urinary bladder cancer. In order to determine amplification levels of all known genes inside the 1.6 kb core amplicon, we constructed a small tissue microarray (TMA) from 9 primary bladder cancers and 4 bladder cancer cell lines with known 6p22 amplification, and analyzed it with a panel of 16 overlapping FISH probes constructed from bacterial artificial chromosomes (BACs). The highest amplification rates were observed for the transcription factor E2F3 and the adjacent gene NM_017774, the function of which is not known. For a more detailed analysis of these genes, additional large section analysis was done in 19 primary bladder cancers and 18 bladder cancer cell lines. It showed that E2F3 and NM 017774 were always coamplified, but amplification levels in terms of the number of gene copies were slightly higher (16-19 copies per nucleus) for E2F3 as compared to NM_017774 (13-15 gene copies). Our study demonstrates that E2F3 and NM_017774 are located on the top of the 6p22.3 amplicon in bladder cancer. It remains to be studied which one of the two genes drives 6p22 amplification, or if both genes contribute jointly to the aggressive features of 6p22 amplified bladder cancers.

Prediction of sudden cardiac arrest: risk stratification by anatomic substrate.

The prognostic importance of coronary artery anatomy to specific outcomes including ventricular tachycardia/fibrillation was evaluated in 372 consecutive patients undergoing cardiac catheterization at University Hospital at Stony Brook between 1981 and 1984. The hypothesis that proximal left anterior descending artery narrowing before the first septal perforator had a specific relationship to survival was again tested in this cohort. The population was prospectively followed for 8 years, with all clinical management decisions made independently by the patient's primary or referring physician. Multivariate statistical and life table analyses were performed after comprehensive follow-up. Significant narrowing in the proximal left anterior descending artery was associated with an increased risk of sudden cardiac death (p = 0.0002). Abnormalities of contractility in the diaphragmatic segment of the left ventricle in addition to an elevation of the left ventricular end-diastolic pressure and the presence of congestive heart failure (p < 0.05) were other contributory variables. Outcome in patients with proximal left anterior descending coronary artery disease who underwent aortocoronary artery bypass to the artery demonstrated improved survival (p < 0.05). Risk stratification of patients at high risk for sudden cardiac death is possible and may allow identification for an aggressive approach or interventional trials.

DNA aberrations in urinary bladder cancer detected by flow cytometry and FISH.

Detection of molecular alterations is of potential significance for diagnosis and prognosis in bladder cancer. Fluorescence in situ hybridization (FISH) allows visualization and quantitation of genes and chromosomes on a cell by cell level and can easily be applied to urinary cells. To evaluate the sensitivity of FISH for detection of DNA aberrations in bladder cancer, formalin-fixed tissues of 293 tumors were examined by FISH and flow cytometry (FCM). Centromere probes for the chromosomes X, Y, 1, 7, 9, and 17 were used for FISH analysis. FISH was more sensitive for detection of quantitative DNA aberrations than FCM. An aberration of at least one chromosome was found in 107 of 108 tumors (99%), which were tetraploid, aneuploid, or multiploid, and in 29 of 49 tumors (59%), which were diploid, by FCM. The frequency of FISH aberrations showed greater differences between pTa (47%) and pT1 tumors (85%; P < 0.0001) than between stages pT1 and pT2-4 (98%). The marked genetic difference between pTa and pT1 tumors argues against the concept of grouping pTa and pT1 tumors together as "superficial bladder cancer." The frequency of tumors with chromosomal aberrations detected by FISH increased with the number of chromosomes examined. Aneusomy was seen in 68% of grade 1 tumors examined for > or = 4 chromosomes, suggesting that the cytological diagnosis of bladder cancer recurrences could be substantially improved by FISH.