The photoreactive carbohydrate probe, a novel method for detection of lectins.

One of the main difficulties in the research of lectins is the absence of an adequate technique for their specific and routine detection. Here, we present a photoreactive carbohydrate-probe which could help to overcome this problem. The probe was comopsed by joining four segments: (i) a carbohydrate moiety, (ii) the digoxigenin tag, (iii) the photoreactive cross-linker and (iv) the lysyl-lysine backbone. After incubation with lectins in the dark, the probe can be activated and cross-linked to the lectins by illumination. The result is a lectin with covalently incorporated digoxigenin tag. Such a labelled lectin can be easily identified using anti-digoxigenin antibodies in a Western blot technique. This method is of high specificity and sensitivity and enables direct detection of lectins in complex mixtures, even whole cell homogenates.

Cytological and biochemical methods for the characterization of in vitro cultured cells.

The aim of our presentation was to show how we characterize cells cultured in monolayer system. Cytological and biochemical methods were used. Ovarian Krukenberg tumour fibroblasts were investigated and findings were correlated with normal human diploids (HDZ1) and with fibroblasts obtained from Blighted ovum. Cytomorphologically Malignancy associated changes in the tumour fibroblasts were found. Cytochemically acid phosphatase and alpha-naphtyl-esterase were positive (+++). PAS reaction was doubled in 18th passage. Cytogenetically normal human diploids were found. Biochemically enzymatic assay showed phosphopentose shunt is decreased in tumour fibroblasts and alpha-glucosidase and beta-galactosidase activities were significantly lower in these cells. A form of N-acetyl-beta-glucosaminidase fell during the investigation from normal 75% to lower percent (42% of the total activity). Much more parameters were obtained by different methods and Krukenberg tumour fibroblasts may be better understood. In vitro investigation makes a contribution to biomedical knowledge in cancer research.