Human carcinoma cell lines xenografted in athymic mice: biological and antigenic characteristics of an intraabdominal model

Summary In order to investigate in vivo clinical applications of murine monoclonal antibodies directed against human ovarian carcinoma a preclinical in vivo model was developed using BALB/c athymic mice. Three human carcinoma cell lines (MCF7, HT29, and SW626) were injected into the peritoneal cavity of pristane-primed animals and the biological and antigenic characteristics of the i.p. grown tumors were studied. The animals were killed when moribund or 6–8 weeks after tumor injection. At autopsy tumor take was observed in 85% of the injected animals, whereas palpable nodules were evident in only 83%. Examination of the peritoneal cavity revealed intraabdominal carcinomatosis with tumor masses varying in size between 0.2 and 0.5 cm in diameter and tumor sheets. The most frequently affected organs were the diaphragm, the liver, and the reproductive system. Ascitic fluid formation was rare and no animal developed tumors outside the peritoneal cavity. To determine whether the in vivo tumors retained the same antigenic characteristics as the in vitro cell lines, four monoclonal antibodies (MBrl, MOv2, MOv8, and MOv15) directed against ovarian carcinoma-associated antigens and two different experimental approaches (immunofluorescence and immunoblotting) were used. Variations at either a quantitative or a qualitative level were observed for some antigens, whereas no evident changes were apparent for others. In particular, the antigens detected by MBr1 and MOv15 on the MCF7 line both maintained high levels of expression and immunoblotting staining pattern, whereas the antigens detected by MOv2 on the HT29 and SW626 lines, although present at a high level, clearly changed their staining pattern. As regards the antigens recognized by MOv8 and MOv15 on the HT29 and SW626 lines, we observed a drastic decrease in the level of their expression and in many cases a drop below the threshold of detectability of the test. The intraabdominal carcinomatosis described partially mimics the growth characteristics of human ovarian cancer and maintains the expression of some antigenic markers associated with epithelial tumors of the ovary and may therefore be useful in devising immunodiagnostic and/or immunotherapeutic strategies for ovarian carcinoma.     

Ultrastructural immuno-localization of tropoelastin in the chick eye

Summary Immuno-gold labeling at the electron-microscopy level was used to investigate the distribution of tropoelastin in the chick eye. Intense staining was found in the amorphous part of mature elastic fibers in different regions of the organ. In elaunin fibers, both the amorphous core and the surrounding microfibrils were clearly labeled. In addition, reactive sites were detected in the oxitalan fibers of the stroma of the cornea and in Descemet's membrane, which showed a gradient of reactive sites increasing from the center toward the periphery. Oxitalan fibers of the stroma often fused with Descemet's membrane; the pattern of immunological staining suggested a continuity between the two structures. In the ciliary zonule, labeling for tropoelastin was observed in discrete areas on the bundles of microfibrils. The results show a complex structural organization of elastic tissue; this may be important in endowing the various parts of the eye with different mechanical properties.     

Factors affecting the survival of E. coli in a river

The relationships between physical, chemical and microbial characteristics of an aquatic ecosystem and the survival of E. coli have been studied. Two conditions of the ecosystem (warm and cold) are considered. T₉₀ (time necessary for 90% of a bacterial population to die) in the warm situation shows an inverse exponential relationship with water temperature. Besides the direct relationship temperature-T₉₀, there is an indirect effect of temperature upon T₉₀ through the natural microflora of the water. The relationships between temperature and the heterotrophic population, and between the heterotrophic population and the bacterial consumers (P.F.U.), are exponential and linear, respectively.     

Trophic role of rotifers in the plankton of lake Kozjak (Plitvice Lakes)

The trophic role of rotifers in the zooplankton community of dimictic, oligotrophic lake Kozjak, the largest lake of the Plitvice Lakes, NW Dinarid Mountains, is analyzed. Their spatial and temporal biomass distribution in relation to that of protozoans, cladocerans and copepods shows that they form a significant part of the non-predatory zooplankton of this karstic standing water.     

Improved high-affinity binding of 3H-apomorphine with a subcellular membrane fraction of mammalian brain tissue

The binding of low concentrations of [³H](?)apomorphine to preparations of calf and rat forebrain tissue was evaluated. Fractionation of crude homogenates to prepare a membrane fraction (P₄) of striatal or caudate homogenates increased the proportion of saturable to total binding from 33% to over 80%, and increased the apparent density of binding sites from 94 to 681 fmol/mg protein. Binding in calf caudate P₄ tissue was protein-dependent and optimal at pH = 7.0 to 7.5, and T = 20 to 25°C; at higher temperatures tissue binding sites appeared to degrade. The half-time of association and dissociation at 22°C were, respectively, 14.0 and 18.5 min; equilibration was complete in 60 min. Kinetic characteristics of high-affinity binding obtained from association and dissociation constants and from saturation isotherms were similar (K_d = 2.1 to 3.4 nM). The pharmacology of competition for ³H-APO suggests selectivity for dopamine-agonist interactions. These results indicate that the P₄ membrane preparation may be useful for the evaluation of dopamine-agonist binding sites or “receptors.”     

Cellulolytic enzymes for the hydrolysis of leached beet cosette

Summary Cellulolytic fungi were isolated from rotting leaves and tested for extra-cellular cellulase activities (CMCase, avicelase, cellobiase and xylanase). The effect of the proportion of the enzyme activities on the rate of degradation of leached beet cosette was observed using a range of supernatant fluids in appropriate combinations. At low cosette concentrations (1.5–3.0 g/l), avicelase and cellobiase were the rate limiting enzymes; avicelase in the initial stages of reaction and cellobiase after 6–8 hours, when cellobiose inhibition becomes important. A ratio of celiobiase to avicelase of approx 2.0 was established as appropriate. At higher substrate concentrations (10 g/l, 40 g/l) the best cellobiase to avicelase ratio was maintained and up to 40% hydrolysis was obtained in the 10 g/l incubation with 10 Uav/l and 20 Ucellob/l. At 100 g/l cosette concentration, substrate inhibition was observed.