Postnatal development of amino acid metabolism enzymes in the liver and muscle of 'cafeteria' rats

The effect of feeding a high-energy highly palatable cafeteria diet on the liver and muscle ontogenesis of serine dehydratase, alanine transaminase, glutamine synthetase and adenylate deaminase during postnatal development of the rat has been studied. The results are in agreement with the lower amino acid utilization in cafeteria rats, both adults and during postnatal development. The feeding of excess energy coupled with high-quality protein resulted in changes in the ontogenesis of the studied enzymes that coincide with the development of protein synthesis and overall pup growth even before they had direct access to this rich diet, suggesting that cafeteria feeding already affects the amino acid metabolism of the pup through the dam's milk.     

Urinary catecholamine responses to basic types of physical activity

Urinary adrenaline, noradrenaline, heart rate, and subjective ratings were obtained from 9 healthy males during six different physical activities, ranging in intensity from lying down to running. Heart rate, subjective ratings and noradrenaline excretion reflected the work load in the different conditions. Adrenaline, on the other hand, failed to show this relationship. There was no significant increase in adrenaline excretion even at the highest work load (corresponding to a heart rate of 160 bpm). It was concluded that urinary adrenaline may safely be used as an indicator of mental factors even in situations with different levels of physical activity.     

Immobilization of lactate dehydrogenase on polyacrylamide beads

Pig muscle lactate dehydrogenase (L-lactate:NAD oxidoreductase, EC 1.1.1.27) was covalently immobilized on polyacrylamide beads containing carboxylic functional groups activated by water-soluble carbodiimide. The effects of immobilization on the catalytic properties and stability of the lactate dehydrogenase were studied. There was no shift in the pH optimum of the immobilized enzyme compared to that of the soluble one. The apparent optimum temperature of the soluble enzyme was 65 degrees C, while that of the immobilized enzyme was between 50 and 65 degrees C. The apparent Km values of the immobilized enzyme with pyruvate and NADH substrates were higher than those of the soluble enzyme. As a result of immobilization, enhanced stabilities were found against heat treatment, changes in pH, and urea denaturation.     

Trypanosoma cruzi: distribution of fluorescently labeled tubulin and actin in epimastigotes

Cytoskeletal components were visualized in epimastigote forms of Trypanosoma cruzi by double immunofluorescence microscopy using monospecific antibodies against tubulin and against actin. Intense staining of the flagellum and the edges of the cell body was observed when the cells were stained with anti-tubulin, reflecting the presence of the basal bodies, the flagellar axoneme and the subpellicular microtubules. A less intense staining was seen in the cell body of epimastigotes stained with anti-actin. However, an intense staining was observed with this antibody in the flagellum, in a pattern similar to that observed with anti-tubulin. It is suggested that the paraxial structure, which is formed by a complex array of 6-nm-thick microfilaments is composed, at least in part, of actin.     

Desangeloylshairidin,X-ray structure determination of a sesquiterpene lactone from Guillonea scabra

The structure and absolute configuration of desangeloylshairidin, a guaianolide isolated from Guillonea scabra, have been established by X-ray diffraction analysis. No conformational change was observed in its seven-membered ring between the crystal and deuterochloroform solution states.     

Changes with starvation in the rat of the lipoprotein lipase activity and hydrolysis of triacylglycerols from triacylglycerol-rich lipoproteins in adipose tissue preparations.

Lipoprotein lipase activity was higher in fat-pad pieces than in isolated adipocytes from the same fed rats, whereas hydrolysis of triacylglycerols from triacylglycerol-rich lipoproteins was similar in the two preparations when incubated either in basal conditions or in the presence of heparin. In both preparations there was a similar release of lipoprotein lipase activity into the medium during basal incubation, enhanced by the presence of heparin. In fat-pad pieces, but not in isolated adipocytes, incubation with heparin produced a decrease in the lipoprotein lipase activity measured in the tissue preparation. In fat-pad pieces from 24 h-starved rats, lipoprotein lipase activity was the same as in isolated adipocytes from the same animals and incubation with heparin did not affect the appearance of lipoprotein lipase in the medium or the utilization of triacylglycerols from triacylglycerol-rich lipoproteins. These results support the following conclusions. (1) The effectiveness of lipoprotein lipase in adipose tissue preparations in vitro depends more on its availability to the substrate than on its total activity. (2) Heparin acts on adipose tissue preparations from fed animals both by enhancing the release of pre-existing extracellular enzyme (which is absent in isolated adipocytes) and by enhancing the transfer outside the cells of the intracellular (and mainly undetectable) enzyme that is activated in the secretion process. (3) In adipose tissue from starved animals there is not only a decrease in the active extracellular form of lipoprotein lipase activity but also a reduction in the intracellular (and mainly undetectable) pool of the enzyme.     

Demonstration of an ATPase at the cell surface of intact normal and neoplastic human cells in culture

An ATPase reaction has been studied at the surface of intact normal and neoplastic human cells in culture. For this purpose a sensitive method has been developed which permits repeated monitoring of enzyme activity without interference with cellular viability. Experiments could be performed with the cells cultured in a single dish. The cells were firmly attached to the supporting medium throughout the experiment. The incubation medium could easily be separated from the cells at the end of the reaction. There was no diffusion of the surface-located ATPase into the incubation medium. Another advantage was the possibility of microscopic control of the appearance of the cells during the reaction. High ATPase activity was found in glia-like cells derived from normal adult brain cells while lines from gliomas had very low activity. One SV40 transformed glia line had an extremely low ATPase activity in contrast to the uninfected cells. Normal fibroblasts and sarcoma cells had about the same low activity as the glioma cells.     

Steroid metabolism in primates. XV. Excretion of C19 and C21 steroids in the urine of baboons (Papio hamadryas) after long-term treatment with a combination of mestranol-chlormadinone acetate

The urinary excretion of C21- and C19-steroids was investigated in female babons (Papio hamadryas) treated with the ovulation inhibitor Ovosiston (mestranol + chlormadinone-acetate), in comparison with an untreated control Group Urinary C21-steroid excretion was not significantly altered by Ovosiston. 17-Ketosteroids were decreased, predominantly 11-oxygenated compounds.     

Glucose transport in Entamoeba histolytica

That the uptake of glucose by the parasitic amoeba Entamoeba histolytica occurs by an equilibrative transport system is supported by the following observations. 1. The rate of glucose uptake is several orders of magnitude greater than the uptake by pinocytosis. 2. The uptake of glucose exhibits saturation kinetics, with K(m)=1.6mm and V(max.) ranging from 2 to 5mumol/min per ml of cells at 37 degrees C. 3. The glucose analogues 2-deoxyglucose, 3-O-methylglucose and d-xylose are transported by the glucose system although with much less affinity. Competitive inhibition was observed between pairs of substrates, with K(i) values for any sugar closely coincident with the corresponding K(m). 4. d-Xylose, a sugar not metabolized by the cells, equilibrated with 80% of the amoebal cell water. 5. Cells equilibrated with xylose exhibited countertransport of this sugar against its concentration gradient when another substrate was added to the medium. 6. Blocking of glycolysis by iodoacetate or F(-) has no immediate effect on transport. The presence of a glucose-transport system in E. histolytica contrasts with the situation found in the non-parasitic amoeba, where pinocytosis seems to be the only mechanism of solute uptake.