In vitro maintenance of differentiation marker synthesis by subpopulations of mouse thymocytes

Mouse thymocyte populations enriched in functionally incompetent, “immature” cells on the one hand, or in competent “mature” cells on the other hand, express different steady-state levels of certain surface antigens and marker enzymes. In the cases of the glycoproteins H-2 (K and D), Qa, and TL, and the DNA polymerase terminal deoxynucleotidyl transferase (TdT), these levels reflect different rates of de novo synthesis in the two populations. Thus each population appears to manifest a characteristic pattern of synthetic rates for the various products relative to total protein synthesis. To investigate the maintenance of these patterns, enriched pools of “immature” and “mature” thymocytes were incubated in vitro for 24 h, and the rates of product synthesis before and after culture were compared. H-2 synthesis, initially most rapid in the mature cells, continued to be made at the highest rate in this population. TdT synthesis, a characteristic activity of the immature cells, was not induced in the mature cells, but proceeded at an increased relative rate in the immature population. Therefore, the differences between the rates of H-2 and TdT synthesis were stable properties of the two thymocyte populations. Another marker of immature cells, TL, did not continue to be produced in parallel with TdT. Rather, its synthesis was selectively curtailed in relation to the continuing protein synthesis in the immature cultures. This non-coordinate regulation of TL and TdT production in immature thymocytes may be due to several mechanisms. These are discussed with regard to their implications for pathways of thymocyte maturation.     

Correlation between cell density,membrane fluidity,and the availability of transferrin receptors in friend erythroleukemic cells

Undifferentiated Friend erythroleukemic cells (FL cells) acquire membrane microviscosity ( ), in accord with the culture cell density. At low cell density poise, whereas at confluency it increases to poise. Concomitantly, the total number of available transferrin receptors per cell decreases by about 80% upon increase in cell density. Modulation of membrane microviscosity, by artificial alteration of the membrane cholesterol level, mediates similar modulations of the availability of the transferrin receptors. The correlation between the availability of the transferrin receptors and the membrane lipid fluidity may take part in the overt decrease in iron uptake by erythroid cells along the erythropoiesis pathway.     

Relative addition rate and external concentration; Driving variables used in plant nutrition research

Abstract An increasing literature accounting for various types of experiments indicates that far lower external nutrient concentrations are required by plants than is usually thought to be the case. It is concluded that the ion uptake capacity of the roots, as described by the carrier concept, is high compared to that required for maintenance of the internal concentration. Serious errors in experimental conclusions are associated with insufficient and constant nutrient addition rates. The main errors are caused by non-steady states of the plants both with regard to the internal nutrient concentrations and the relative growth rate. A dynamic concept has been proposed for direct use as the treatment variable within the range of sub-optimum nutrition. The nutritional factor is expressed as a flow, the relative nutrient addition rate in laboratory studies and the nutrient flux density in the field. The experimental use of the relative addition rate has led to steady-state nutrient status and relative growth rate and the interpretation of plant responses which differ fundamentally from accepted views. Thus, for instance, deficiency symptoms disappear, as in natural conditions, when the internal nitrogen concentration is stable, independent of level. The nutrition/growth relationships are very different from those observed when external concentration is varied. The regression line of relative growth rate on relative addition rate passes near to the origin at an angle close to 45 to the axes, which implies that the obtained relative growth rate approximates closely the treatment variable. A striking example of observed differences is the positive effect on nitrogen fixation exerted by high relative nitrogen addition rates compared to the well-known negative effect of increasing external nitrogen concentration. The application of fertilizer on the basis of the nutrient flux density concept provides the possibility of supplying fertilizers corresponding to the consumption potential of the vegetation and to the natural flux density resulting from mineralization in the soil. Nitrogen utilization is high under such conditions and the resulting feedback of nutrition on the mineralization rate suggests that there will be a long-term increase in fertility.     

Evaluation of the potential of a bead mill for the release of intracellular bacterial enzymes

The suitability of bead mills for the release of intracellular bacterial enzymes has been studied using the Dyno-Mill Model KDL. The effect of cell concentration, bead size and agitation speed on the release of beta-lactamase from Enterobacter cloacae P99 was examined. Scale-up considerations included, the best operational values for these parameters were 1 g cell paste suspended in 2.5 ml buffer, 0.25 mm diameter glass beads and 15 ms ⁻¹ agitation speed. These conditions proved suitable for the release of enzymes from other Gram-negative bacteria in both batch and continuous processes.     

Synthesis of two forms of apolipoprotein B by cultured rat hepatocytes

Cultured rat hepatocytes were used to demonstrate that the liver can synthesize two forms of apolipoprotein B. Separation of apolipoprotein B by disc gel electrophoresis indicated that hepatocyte low density lipoprotein contains predominantly apolipoprotein B with an apparent molecular weight of 345,000 ± 5,055. In contrast, the major apolipoprotein B component of hepatocyte very low density lipoprotein is a variant form with a molecular weight of 242,000 ± 2,720. Hepatocyte high density lipoprotein, unlike plasma HDL, also contains apolipoprotein B with an apparent molecular weight of 244,000 ± 2,742. Incorporation of [³H] leucine into hepatocyte apolipoprotein B components suggested de novo synthesis.     

Population ecology of Ephydra cinerea Jones (Diptera: Ephydridae), the only benthic metazoan of the Great Salt Lake,U.S.A.

The population of Ephydra cinerea was studied during three summers when dissolved solids levels were about 130 g/l. All life stages are present year-round, but there is apparently some coordination of the initial pulse of adult emergence in June. There are probably 1–2 generations per year.Eggs are deposited on the water surface; inputs to three locations were similar. Larvae successfully avoid the large benthic area covered by an anaerobic monimolimnion. They tend to immigrate from substrates where they grow relatively poorly, and to remain on substrates where they grow better. Consequently, larval densities are more than ten times higher on reef and shallow water mud substrates than on sand. This marked spatial specialization in the absence of substrate-specific predators or competitors illustrates the power of habitat quality by itself in determining spatial patterns of abundance in a lake.Experiments showed larval growth on the reef was inversely related to density, and the lake as a whole produced relatively larger flies in a year when larval and pupal densities were relatively low. Yearly production by E. cinerea is roughly 50 g/m², about 88% of which comes from reefs and shallow water mud areas covering only 18% of the bottom area.Past studies indicate that blue-green algae dominate the lake's benthic flora when salt concentrations are high (due to low lake levels), and diatoms take over when salt concentrations are low. Fly abundance appears to be inversely related to salinity.The lake's present high planktonic primary production is equal to that of eutrophic freshwater lakes, yet it has water clarity more characteristic of an oligotrophic lake. The high water clarity (which makes possible the high benthic production) probably depends on the absence of phytoplankters that can both tolerate the high salinity and avoid being eaten by Artemia salina. Continued dilution of the lake will probably upset this situation and result in reorganization of the lake's energy flow pattern.     

Distribution of chylomicron degradation products in the isolated,perfused rat heart

Chylomicron degradation by hearts from fed and fasted rats was studied using a perfusion technique, which allows the separate collection of coronary (Q_rv) and interstitial effluent (Q_i). Upon perfusion with [³H]-cholesterol-containing chylomicrons the tissue recovery of label was highest in the fasted state, while label recovered in Q_i was highest in the fed state. Density gradient centrifugation of Q_i indicated that the label was recovered in lipoproteins with higher densities: low density lipoproteins (1.019<d<1.050), high density lipoproteins (1.050<d<1.21) and a fraction of d>1.21. These particles probably represent chylomicron degradation products (remnants and “surface fragments”). Our results indicate that tissue cholesterol uptake during chylomicron degradation may be inhibited in the fed state. Furthermore, the role of the myocyte (or interstitial) lipoprotein lipase in chylomicron degradation is discussed.     

Purification and properties of the Bombyx mori nuclear polyhedrosis virus liberated from polyhedra by dissolution with silkworm gut juice

Occluded virions of the Bombyx mori nuclear polyhedrosis virus were efficiently liberated from polyhedra by dissolution with the silkworm gut juice. The liberated virions were purified by sucrose density gradient centrifugation and the bands of enveloped virions were observed in the gradients. There was no functional difference between the gut juice-liberated and the carbonate-liberated virions. Disruption of enveloped virions by the gut juice was observed, but the formation of nucleocapsids from the degradation of the occluded virions was not detected. High yields of the enveloped virions from the polyhedra dissolved by the gut juice was obtained by separating the virions through sucrose density gradient centrifugation immediately after the dissolution of the polyhedra. Many factors, e.g., rearing seasons, silkworm strains, and rearing conditions, affect the polyhedra-dissolving property of the larval gut juice.     

Comparative effects of ethanol, n-propranol and isopropanol on lipid disposal by rat liver.

Besides ethanol, other aliphatic alcohols such as n-propanol and isopropanol induce a triacylglycerol (TAG) accumulation in the liver. To determine whether a common mechanism is responsible for the effects of these three alcohols on hepatic lipid metabolism, each was administered by gastric tube to female Wistar rats at the dose of 50 mmol/kg body wt. Whichever alcohol was administered, the hepatic triacylglycerol accumulation was found to be related to the duration of elevated blood alcohol concentration. After administration of n-propanol or isopropanol, the liver [14C]palmitate uptake was increased whereas hepatic palmitate oxidation to 14CO2 was impaired and palmitate esterification into TAG enhanced; these perturbations were however more discrete than after ethanol administration. In contrast to ethanol and n-propanol which, at the dose presently used, increase precursor incorporation into blood TAG, isopropanol inhibits this incorporation. Interference with the process of very low density lipoprotein (VLDL) synthesis and/or secretion, which appears only at a late stage of isopropanol intoxication, is probably responsible for the intensity and duration of the fatty liver observed after administration of this alcohol.