Covalent immobilization of the multienzyme enniatin synthetase

Summary The multienzyme enniatin synthetase was covalently immobilized to N-hydroxysuccinimide activated agarose. The stability of the immobilized enzyme at 25°C was enhanced compared to the soluble enzyme. Immobilization experiments also indicated that the enniatins are synthesized by a single molecule and thus do not require interactions of several enzyme molecules.     

Production and characterization of KDO-specific monoclonal antibodies recognizing lipopolysaccharides from heptoseless mutants of Salmonella

Abstract Hybrid cell lines producing monoclonal antibodies with specificity for the lipopolysaccharide (LPS) from the deep rough mutant Salmonella minnesota R595 have been established. Spleen cells from BALB/c mice immunized with live R595 bacteria were fused with Sp 2/0 myeloma cells and three hybridomas producing antibodies specific for heptoseless LPS from Salmonella were selected. All three monoclonal antibodies were shown to bind only to heptoseless, but 3-deoxy- d -manno-octulosonic acid (KDO) containing LPS when tested in enzyme-linked immunosorbent assay (ELISA) against a set of structurally defined LPS and lipid A from Salmonella, Shigella and Escherichia coli . Synthetic KDO was an efficient inhibitor of the antibody-R595 LPS interaction defining that KDO is in an immunodeterminant position interacting with the monoclonal antibodies.     

Elevation of the Extracellular Concentrations of Glutamate and Aspartate in Rat Hippocampus During Transient Cerebral Ischemia Monitored by Intracerebral Microdialysis

Rats were implanted with 0.3-mm-diameter dialysis tubing through the hippocampus and subsequently perfused with Ringer's solution at a flow rate of 2 microliter/min. Samples of the perfusate representing the extracellular fluid were collected over 5-min periods and subsequently analyzed for contents of the amino acids glutamate, aspartate, glutamine, taurine, alanine, and serine. Samples were collected before, during, and after a 10-min period of transient complete cerebral ischemia. The extracellular contents of glutamate and aspartate were increased, respectively, eight- and threefold during the ischemic period; the taurine concentration also was increased 2.6-fold. During the same period the extracellular content of glutamine was significantly decreased (to 68% of the control value), whereas the concentrations of alanine and serine did not change significantly during the ischemic period. The concentrations of gamma-aminobutyric acid (GABA) were too low to be measured reliably. It is suggested that the large increase in the content of extracellular glutamate and aspartate in the hippocampus induced by the ischemia may be one of the causal factors in the damage to certain neurons observed after ischemia.     

Dolichol and Dolichyl Phosphate Levels in Brain Tissue from English Setters with Ceroid Lipofuscinosis

Human ceroid lipofuscinosis (CL) is an inherited disease marked by cerebromacular degeneration and early death. We have utilized the canine model to investigate the possible role of dolichol and dolichyl phosphate in the developmental pathology of CL. We found that while brain levels of dolichol increase with age in both affected and unaffected dogs, the amount in the diseased animal was similar to that in controls. Brain levels of dolichyl phosphate ranged from 20 to 35 micrograms/g in control dogs at all ages examined, but increased substantially during development in the affected dogs, a value of 113 +/- 24 micrograms/g (mean +/- SD) being obtained in the end-stage animals. In addition to the results obtained in the canine model, dolichyl phosphate levels in human brain tissues from a 5-year-old with late infantile CL and from a 19-year-old with juvenile CL were found to be 153 and 382 micrograms/g, respectively, compared with a control that assayed 26 micrograms/g. The preliminary findings with human tissues provide further evidence for an association of elevated brain dolichyl phosphate levels with CL. Whether the increase is primary or secondary remains to be determined.     

On the interaction between stabilizing social factors and destabilizing trophic factors in small rodent populations

Mathematical models are developed in order to analyze whether or not social factors, such as, for example, the “social fence” (J. B. Hestbeck, 1982, Oikos 39, 157–163) will stabilize population density: the dynamic interaction between social factors and (dynamic) trophic factors is analyzed. It is concluded that social factors such as the “social fence” tend to stabilize population density; hence, if density cycles (as, e.g., seen in many microtine rodents) are observed in nature, it seems reasonable to conclude that density cycles are driven by, for example, trophic interactions and not by social factors. It is suggested that the “social fence” may explain why so many populations including several microtine populations have fairly stable densities despite the ever-existing destabilizing trophic interactions. Contrary to what is implied by J. B. Hestbeck (1983, “A Mathematical Model of Population Regulation in Cyclic Mammals,” Lecture notes in biomathematics, Vol. 52, Springer-Verlag, Berlin/New York), the analysis presented in this paper demonstrates that seasonal environmental changes are not essential for the generation of regular density cycles. Seasonal changes may, however, be necessary for generating a microtine-like density cycle. Empirical information on microtine rodents relating to the “social fence hypothesis” is discussed.     

Studies of Electrophoresis on Cellulose Acetate Membrane of Serum Proteins from Normal Horses,Sheep and Pigs

A method for the rapid electrophoresis on a cellulose acetate membrane of serum proteins from horses, sheep and pigs is discussed. The various main globulin fractions in the serum of these animals were experimentally identified. Normal values for the percentage composition of serum from normal horses, sheep and pigs were calculated. In the horse there was great individual variation in the shape of the β-fraction, assumed to be due to different transferrin types. The mean value for β-globulin of 19.5 % in the horse was higher than for the other two species. The albumin percentage was highest in the sheep and lowest in the pig, 48.5 % and 43.2 % respectively. The sheep had the highest γ-globulin percentage, 22.8 %, while the horse had the lowest with 19.0 %. Finally the values were compared with corresponding figures reported by other authors and the results discussed.     

Starvation survival and body composition in mammals with particular reference toHomo sapiens

A computer model of body mass and composition in relation to gross energy balance is constructed. The model is built using conventional empirical physiological formulae rather than statistical or analytical mathematical techniques. The model is applied to the Minnesota and other experiments and produces as good or better simulations of observed values of changes in body weight than reported for other formulae or models. Alternative physiological mechanisms concerning metabolic adaptions to starvation, changes in time activity budgets and the energy equivalents of weight loss offer equally good simulations of experimental results. The present analysis highlights the survival value of a basal metabolic depression during starvation and indicates an optimal body composition of 10% mobilizable fat for starvation survival for a 70 kg man. Proper quantification of the effects of the physiological mechanisms involved depends on new experimental data, however. Long term continuous monitoring of time activity budgets are a necessary part of such experiments.