The use of fixed-frozen tissue in routine ultrastructural evaluation and immunogold labeling.

A method of freezing fixed surgical tissue specimens for long-term storage and subsequent resin embedding was developed. The method entails fixation of the specimens in Karnowsky's liquid, cryoprotecting with 10% sucrose, freezing in melting Freon gas, and storage at -80 degrees C. When needed, even after years, the frozen tissue pieces are thawed in lukewarm (37 degrees C) buffer and embedded in resin (Epon or Lowicryl). Ultrastructural detail as well as immunocytochemical reactivity of a wide variety of cellular constituents are well preserved. The method is practical and allows retrospective embedding in Lowicryl resin (a technique not easily applicable in everyday routine) for subsequent ultrastructural immunolocalization studies.

[Intravesical chemoprophylaxis with beta-interferon in superficial carcinoma of the bladder]. / Chemioprofilassi endovescicale con beta-interferone nel carcinoma superficiale della vescica.

24 patients with recurrent (group 1) or primary (group 2) bladder cancer (ta/t1 G2) were given 50.000.000 U.I. of beta interferon intravesically after TUR of the neoplasm. In group 1 recurrence rate (rr) was 100% in the period before INF and 83% after INF treatment (follow-up 15-24 months, median 19.8) (p = ns). In group 2 rr was 25% in a 24 months, median 19.8) (p = ns). In group 2 rr was 25% in a 24 months follow-up period; if compared to the rr of a historical control group treated by only a TUR (38.2%) there was a statistically significant difference (p less than 0.01). Tolerability of the drug was excellent.

Ultrastructural cytoskeleton alterations and modification of actin expression in the NIH/3T3 cell line after transformation with Ha-ras-activated oncogene.

Cytoskeleton alterations of NIH/3T3 fibroblast monolayers transfected with Ha-ras-activated oncogene were studied by immunofluorescence, immunoelectron microscopy, and immunoelectrophoretic analysis of actin isoforms. Transformation foci were found to consist of cells with a round shape and rare stress fibers that spread sparsely, forming rare focal contacts and fibronexuses. The loss of stress fibers in transformed cells was confirmed by staining with rhodamine-phalloidin and with a fluorescinated anti-non-muscle cell actin antibody. The transformed cells were anchored to the substrate prominently by filaments that contained fibronectin, as showed by immunoelectron microscopy. A down-regulation of alpha-actin isoform was observed by immunofluorescence and immunoblotting analysis using a specific monoclonal antibody. The diffuse distribution of alpha-actin, lacking a specific association with stress fibers, challenges the hypothesis of a connection between alpha-actin down-regulation and stress fiber loss.

Serum and urine ferritin in patients with transitional cell carcinoma of the bladder.

Erythrocyte, serum and urine ferritin concentrations were evaluated in 20 patients suffering from transitional cell carcinoma of the urinary bladder and in 20 healthy men. No clinical or biochemical signs of liver disorders, chronic inflammatory states or infections were present in both the patients and the controls. Our results showed no significant difference in the erythrocyte ferritin concentration in both groups. On the contrary there was a statistically significant difference in mean serum (p less than 0.05) and urine (p less than 0.01) ferritin concentration between the two groups. The mean serum ferritin concentration in the patients was 102.23 +/- 63.38 ng/ml while it was 258.41 + 250.68 ng/ml in normal subjects. The mean urine ferritin concentration was 6.30 +/- 5.35 ng/ml in normal subjects and 22.66 +/- 25.59 ng/ml in patients with bladder cancer. Our data seem to demonstrate that the assessment of the ferritin either in the serum or preferably, in the urine, could become an interesting tumoral marker for bladder cancer.