Overexpression of phospholipase C-gamma1 in rat 3Y1 fibroblast cells leads to malignant transformation

Phospholipase C-gamma1 (PLC-gamma1) mediates signals from various extracellular origins to evoke cellular events such as mitogenesis. Previously, we reported that PLC-gamma1 was highly expressed in colorectal cancer and familial adenomatous polyposis, suggesting that PLC-gamma1 might be oncogenic. In this study, we have established rat 3Y1 fibroblasts that overexpress whole PLC-gamma1 and src homology 2 (SH2)-SH2-SH3 domain of PLC-gamma1. These cells showed a transformed phenotype and were tumorigenic when transplanted into nude mice. These results indicate that overexpression of PLC-gamma1 could transform rat fibroblasts, and the transformation is mediated by SH2-SH2-SH3 domain of PLC-gamma1.     

Antibody catalysis of peptidyl-prolyl cis-trans isomerization in the folding of RNase T1

An antibody generated to an alpha-keto amide containing hapten 1 catalyzes the cis-trans isomerization of peptidyl-prolyl amide bonds in peptides and in the protein RNase T1. The antibody-catalyzed peptide isomerization reaction showed saturation kinetics for the cis-substrate, Suc-Ala-Ala-Pro-Phe-pNA, with a kcat/Km value of 883 s-1.M-1; the reaction was inhibited by the hapten analog 13 (Ki = 3. 0 +/- 0.4 microM). Refolding of denatured RNase T1 to its native conformation also was catalyzed by the antibody, with the antibody-catalyzed folding reaction inhibitable both by the hapten 1 and hapten analog 13. These results demonstrate that antibodies can catalyze conformational changes in protein structure, a transformation involved in many cellular processes.     

Human melanocytes and keratinocytes exposed to UVB or UVA in vivo show comparable levels of thymine dimers

Epidemiology shows a relationship between solar exposure and all types of skin cancer. Understanding the mechanisms of skin cancer requires knowledge of the photomolecular events that occur within the relevant epidermal cell types in vivo. Studies to date have focused on UVR-induced DNA lesions in keratinocytes, the majority epidermal cell population which gives rise to most skin cancers. Malignant melanoma, arising from melanocytes (5%-10% of epidermal cells), accounts for most skin cancer deaths. We report on new techniques to detect DNA photolesions in human epidermal melanocytes in situ. Previously nonexposed buttock skin of volunteers of skin types I/II was exposed to clinically relevant doses of narrow bandwidth UVB (300 nm) and UVA (320 nm, 340 nm, 360 nm) radiation. Biopsies were taken immediately afterwards and processed for routine histology. Microscope sections were prepared and double-stained with fluorescent-tagged monoclonal antibodies for thymine dimers and melanocytes. UVR dose-response curves for dimer levels within melanocyte nuclei were determined by image analysis and compared with dimer levels in adjacent basal cell keratinocytes. Our data show that UVB and UVA readily induce thymine dimers in melanocytes at levels that are comparable with those found in adjacent keratinocytes. This new technique will enable melanocyte specific studies, such as DNA repair kinetics, to be done in vivo.     

Intravenous long term infusions of the rat without anesthesia

The lumbar-spine radiographs of 67 patients with idiopathic chondrocalcinosis articularis were reviewed for disc calcification and other changes. Calcification was present in 21 (31%) of the patients, seen most frequently at the L 2-3 disc space. The 21 patients as a group were significantly older than the 46 patients without disc calcification, and also had a much higher incidence of chondrocalcinosis in peripheral joints. There was no association with back pain or spinal stiffness. The 21 patients with disc calcification included six patients with a destructive peripheral arthropathy, and three of them had destructive changes affecting the lumbar spine. Three patients with a destructive peripheral arthropathy were also included in the group without disc calcification, and one of these had a destructive arthritis of the lumbar spine. For comparison, there was a 55% incidence of spinal chondrocalcinosis in nine patients with primary hyperparathyroidism and peripheral joint chondrocalcinosis, and a 6% incidence in 100 anteroposterior lumbar spine radiograph 'controls', taken before intravenous urography, although in this latter group the changes were minimal and confined to the margin of a single disc in each case.     

Histometric study of meat products. A literature review

In the present paper, the literature on the development of methods for histometric monitoring the quality of meats is reviewed. The value of specific techniques, statistical interpretation of results and the practicability of automation of histometric analysis are examined more closely. It is concluded from the study of the literature that histometric examination of meats allows an objective assessment of volume percentages of tissue components. When a distinct difference in contrast between various tissues is achieved by specific staining methods, developments in the field of image analysis systems will allow automation of the quantitative histological examination of meats.