Neurons are replenished in cultures of embryonic chick optic tectum after immunomagnetic depletion

The effect of intercellular interactions on the determination and differentiation of early embryonic brain cells was tested by immunomagnetic cell separation techniques. Using the A2B5 monoclonal antibody, which in chick brain reacts with a neuron-specific surface ganglioside, we produced initially pure populations of optic tectum cells devoid of the antigen. A coincident depletion of neurofilament(+) cells (95%) and nonneuronal growth characteristics of the separated A2B5(-) cells indicated that the vast majority of neurons had been removed initially. Surprisingly, A2B5(+) cells rapidly appeared in separated A2B5(-) cell cultures. After 1 day, the percentage of A2B5(+) cells in separated cell cultures equalled those in unseparated cultures (approximately 50%). By a week in culture, A2B5(+) cells developed neuronal morphology and contained neurofilaments. A2B5(-) to (+) conversion was a regulated phenomenon in that removal of different proportions of the (+) cells resulted in different numbers of (-) to (+) conversions. New DNA synthesis was not required for the acquisition of cell surface A2B5 antigen or for differentiation of cells into definitive A2B5(+) neurons. Our results demonstrate that postmitotic embryonic brain contains cells which are capable of replacing depleted neurons in vitro.     

Plasmid stability in a recombinant mammalian cell bioprocess

Summary A simple experimental method is devised to determine the fraction of plasmid-harboring cells in a bioprocess employing recombinant mammalian cells. The fraction of plasmid-harboring cells decreased as serum content in the growth medium decreased. The relatively higher increase in the generation time of the plasmid-harboring cell was primarily responsible for this decrease. The mathematical expression obtained for this fraction in terms of the two parameters, i.e. the generation time ratio and the plasmid-loss probability, could represent the experimental data extremely well. The numerical values of these parameters could show the inherent insight of the system. It was found that the data plot against time can draw us to a misleading conclusion of the absence of the effect of serum concentration.     

Intraspecific resource partitioning in a headwaters stream fish,the pearl dace Semotilus margarita (Cyprinidae)

Synopsis Pearl dace, Semotilus margarita, are common in slow-moving channels and pools of the headwaters of the Brokenhead River. From May to September age groups 0, 1 and 2+ were partially segregated in space based on water depth with age 0 occupying shallow pools and shallow channels. Age 1 were abundant in shallow pools and deep channels while age 2+ occurred in deep channels and deep pools. In November all age groups coexisted in deep pools. Pearl dace are omnivorous consuming invertebrates, plant material, and detritus. From May to September age groups 0 and 1 consumed mainly terrestrial invertebrates (Diptera, Hymenoptera, Thysanoptera) but age 1 consumed more detritus than age 0. Age 2+ consumed mainly aquatic Diptera larvae. Although all ages consumed aquatic insects almost exclusively in November differences in diet between age groups still existed. Younger fish fed higher in the water column than older fish. Intraspecific resource partitioning of food and space occurs between age groups and this combined with the omnivorous diet is highly adaptive to the headwaters of streams.     

Immunoassay using 125I- or enzyme-labeled protein A and antigen-coated tubes

Antigen-coated plastic tubes were used with ¹²⁵I- or enzyme-labeled staphylococcal protein A in a general immunoassay method for antigens and haptens. Protein A reacts with immunoglobulin G (IgG) regardless of antibody specificity at sites distal to the antigen combining site and does not inhibit the immune reaction. It therefore serves as a general tracer and its use eliminates the need to purify and to label individual components for each assay. Macromolecular antigens were bound to polystyrene or polypropylene tubes by direct passive adsorption. Haptens with free carboxyl groups were bound covalently to poly-l-lysine and these conjugates passively adsorbed to the tube surface. Optimal assay conditions were established for the quantitative determination of immunoglobulins and the folate derivatives, methotrexate and 5-methyltetrahydrofolate, using ¹²⁵I-labeled protein A or protein A labeled with alkaline phosphatase. The method has been used to estimate levels of IgG, IgA, IgM, and IgE in serum in volumes up to 1 ml.     

Trypanosoma vivax: Courses of infection with three stabilates in inbred mouse strains

The courses of infection in inbred mouse strains were compared following infection with three Stabilates of high, intermediate, and low virulence of Trypanosoma vivax stock Zaria Y486. Mouse strains could only be shown to differ in their resistance to T. vivax infections as judged by the height of the initial parasitemia and survival times when a trypanosome population of low or intermediate virulence was used. A T. vivax population of high virulence was uniformly lethal. Comparison of lytic antibody titers between groups of resistant ($\text{C}\text{57}\text{B}\text{1}\text{6}$) and susceptible ($\text{Balb}\text{c}$) mice did not show any significant differences in titers of the surviving mice but the mice in either group which did not control the initial parasitemia had lower lytic antibody titers than those which did. A significantly larger number of $\text{Balb}\text{c}$ mice failed to control the initial infection as compared to the $\text{C}\text{57}\text{B}\text{1}\text{6}$. Treatment with cyclophosphamide did not ablate differences in susceptibility between the two strains. The use of congenic mice showed that these differences in susceptibility were not related to differences in the major histocompatibility complex between these strains.     

The effect of exhaustive exercise on expired pentane as a measure of in vivo lipid peroxidation in the rat

The concentrations of pentane and ethane in the expired breath of swimming rats were used to determine the possible occurrence of lipid peroxidation caused by strenuous exercise. Rats swum to exhaustion produced significantly more pentane but not ethane than did rats at rest. Rats fed a vitamin E-deficient diet produced slightly more pentane following exhaustive exercise than they did while at rest, but this increase was not statistically significant. Rats were also swum for prescribed lengths of time. Only rats that had swum for 20 or 40 min had significantly elevated concentrations of pentane in the breath. Rats swum for 10 or 30 min had elevated concentrations of pentane in the breath, but these increases were not statistically significant. These results suggest that lipid peroxidation is moderately increased following exhaustive exercise.     

Amino Acid Synthesis in Photosynthesizing Spinach Cells : EFFECTS OF AMMONIA ON POOL SIZES AND RATES OF LABELING FROM CO(2)

Isolated cells from leaves of Spinacia oleracea have been maintained in a state capable of high rates of photosynthetic CO₂ fixation for more than 60 hours. The incorporation of ¹⁴CO₂ under saturating CO₂ conditions into carbohydrates, carboxylic acids, and amino acids, and the effect of ammonia on this incorporation have been studied. Total incorporation, specific radioactivity, and pool size have been determined as a function of time for most of the protein amino acids and for γ-aminobutyric acid. The measurements of specific radio-activities and of the approaches to ¹⁴C “saturation” of some amino acids indicate the presence and relative sizes of metabolically active and passive pools of these amino acids.     

Molecular characterization of pig muscle phosphoglucose isomerase

As a corollary to X-ray crystallographic work performed by H. Muirhead, detailed studies on crystalline pig muscle phosphoglucose isomerase have been conducted to establish its basic physical and chemical properties. The enzyme species being investigated by X-ray diffraction has been determined to be isoenzyme III. Its molecular weight in the native state was found to be 132,000, its s⁰₂₀,w value to be 7·25 S. The enzyme is composed of two subunits of equal molecular weight (66,000). Its amino acid composition is largely similar to that of rabbit muscle phosphoglucose isomerase, with the significant exception that the pig muscle isomerase contains only three sulfhydryl groups per polypeptide chain (two of them accessible to titration with p-mercuribenzoate) as compared with twice that number for the rabbit muscle enzyme. This low number of sulfhydryl groups is interpreted as being responsible for the ease with which heavy-atom, isomorphous derivatives could be prepared for the pig muscle enzyme by Shaw & Muirhead (1977).     

Comparison of fatty acid oxidation in mitochondria and peroxisomes from rat liver

Oxygen uptake with succinate or palmitoyl-CoA as substrates can be measured in rat liver mitochondria that have been isolated by sucrose density gradient centrifugation providing the fractions are diluted with a 30 mM phosphate buffer rather than with an isotonic medium. Separate assay procedures were used to measure peroxisomal and mitochondrial β-oxidation of palmitoyl-CoA in the fractions of a sucrose gradient used to separate these organelles. A preliminary estimate of the ratio of palmitoyl-CoA oxidation by the mitochondrial fraction relative to the surviving peroxisomes from livers of male rats was 3.2.