Endogenous respiration and regulation in a primitive marine fungus

The relationship between the respiration and the presence and utilization of endogenous and exogenous substrates was studied in the non-filamentous obligately marine fungus Thraustochytrium aureum. Using isotopic and manometric methods, it was shown that almost all exogenous glucose is assimilated, whilst almost all the oxygen consumption in the presence of exogenous glucose was due to oxidation of endogenous reserves. In contrast, exogenous glutamate, which cannot serve as the sole carbon source for growth, inhibits respiration of endogenous materials, and is itself rapidly oxidized. The uncoupler 2,4-dinitrophenol stimulates the oxidation of endogenous reserves without affecting the uptake and use of exogenous glucose. These data strongly support the idea of physiologic compartmentation in this organism.     

Synthesis of α- and β-glycosides containing spin labels,as probes for studies of carbohydrate-protein interaction

Nitroxide spin-labeled α-d-glycopyranosides were synthesized in good yield and in a highly stereoselective manner by reaction of per-O-benzyl-α-d-glycopyranosyl bromides with 2,2,6,6-tetramethyl-4-piperidinol under the bromide ion-catalyzed conditions devised by Lemieux etal. After hydrogenolysis, the deblocked intermidiates were oxidized to give the desired, spin-labeled α-d-glycopyranosides. Nitroxide spin-labeled α-d-glycopyranosides, as well as a β-maltoside, were synthesized by standard methods. The synthesis is also described of 2-amino-2-deoxy-d-glucose and -d-galactose derivatives having a spin label at C-2, and of the spin-labeled compound 1-[4-(β-d-galactopyranosyloxy)phenyl]-3-(2,2,6,6-tetramethylpiperidin-1-oxyl-4-yl)-2-thiourea.     

Weather patterns and asthma epidemics in New York City and New Orleans,U.S.A.

Twenty-five years of data on asthmatic attacks in New Orleans (covering approximately 170,000 asthma attacks) have been analyzed to identify asthma epidemic days, defined as days on which an unusually high number of asthmatic individuals had attacks. Similar data covering three years was obtained for New York City. A preliminary examination of detailed meteoroligical data revealed a consistent meteorological pattern preceding and associated with such asthma epidemic days which consisted of a cold front preceding an asthma epidemic by one to three days followed by a high pressure system. The significance of these meteorological findings and their relationship to other environmental agents such as natural or man-made atmospheric pollutants that are likely to be associated with asthma attacks will be discussed.Presented at the Eighth International Congress of Biometeorology, 9–14 September 1979, Shefayim, Israel.     

The estradiol-stimulated lipoprotein receptor of rat liver. A binding site that membrane mediates the uptake of rat lipoproteins containing apoproteins B and E

Hepatic catabolism of lipoproteins containing apolipoproteins B or E is enhanced in rats treated with pharmacologic doses of 17 alpha-ethinyl estradiol. Liver membranes prepared from these rats exhibit an increased number of receptor sites that bind 125I-labeled human low density lipoproteins (LDL) in vitro. In the present studies, this estradiol-stimulated hepatic receptor was shown to recognize the following rat lipoproteins: LDL, very low density lipoproteins obtained from liver perfusates (hepatic VLDL), and VLDL-remnants prepared by intravenous injection of hepatic VLDL into functionally eviscerated rats. The receptor also recognized synthetic lamellar complexes of lecithin and rat apoprotein E as well as canine high density lipoproteins containing apoprotein E (apo E-HDLc). It did not recognize human HDL or rat HDL deficient in apoprotein E. Much smaller amounts of this high affinity binding site were also found on liver membranes from untreated rats, the number of such sites increasing more than 10-fold after the animals were treated with estradiol. Each of the rat lipoproteins recognized by this receptor was taken up more rapidly by perfused livers from estrogen-treated rats. In addition, enrichment of hepatic VLDL with C-apoproteins lowered the ability of these lipoproteins to bind to the estradiol-stimulated receptor and diminished their rate of uptake by the perfused liver of estrogen-treated rats, just as it did in normal rats. The current data indicate that under the influence of pharmacologic doses of estradiol the liver of the rat contains increased amounts of a functional lipoprotein receptor that binds lipoproteins containing apoproteins B and E. This hepatic lipoprotein receptor appears to mediate the uptake and degradation of lipoproteins by the normal liver as well as the liver of estradiol-treated rats. The hepatic receptor bears a close functional resemblance to the LDL receptor previously characterized on extrahepatic cells.     

Modification of cell membrane in variants of chinese hamster cells resistant to abrin

Stable and heritable variants of Chinese hamster ovary (CHO) cells which are resistant to different levels (0.1, 1.0 and 10 μg/ml) of the toxin abrin have been isolated and characterized. The frequency of resistant colonies to abrin was increased with the concentration of a chemical mutagen. There was no effect of cell density or cross-feeding on the recovery of variants. In experiments using fluorescein-labeled abrin and ricin which bind to terminal (non-sialylated) galactose residues of cell-surface oligosaccharides, parental cells exhibited strong binding toward both toxins, whereas no fluorescence was observed in the resistant clones. A fluorescein-conjugated lectin, BS II, which is specific for terminal N-acetyl- -glucosaminyl residues, did not interact with the parental cells, but did with the resistant clones. This suggests that on the surface of resistant cells the number of terminal galactosyl residues of oligosaccharide chains in glycoproteins was reduced, exposing the penultimate N-acetyl- -glucosaminyl residues. The number of available endogenous acceptor sites for galactosyl transferase in the abrin-resistant clones was directly proportional to the degree of resistance. In the presence of great excess of exogenous acceptor, the rates of galactosyl transfer were similar in all the abrin-resistant cell types tested, with levels ranging from 1.4 to 1.7 times parental cell values. Studies with tetraploid cell hybrids reveal that resistance was a recessive trait. Fluctuation analysis showed that abrin resistance occurred in CHO cell populations at a rate of 4−7 × 10⁻⁸/cell/generation. The system may serve as a new marker for quantitative mutagenesis studies.     

Immunoregulatory influence of thymic epithelium and thymopoietin on human B-lymphocyte differentiation

Thymic influence upon immunoregulation of B-lymphocyte differentiation in human bone marrow was investigated. Mononuclear cells isolated from marrow of normal adult volunteers were incubated with thymic epithelial monolayers or with the polypeptide thymopoietin. Generation of pokeweed mitogen-stimulated anti-sheep red blood cell antibody-secreting direct plaque-forming cells (PFC) was found to be inhibited following incubation of marrow mononuclear cells with thymic epithelial monolayers. Addition of 50 ng/ml thymopoietin to pokeweed mitogen-stimulated cultures resulted in enhanced marrow PFC responses, whereas higher doses of thymopoietin were inhibitory for the generation of PFC in this assay system. The data suggest that both helper and suppressor T cells are recruited from their precursors in human bone marrow by thymic influences. Generation of helper or suppressor cells may be dependent upon (a) the stage of differentation of precursor T cells and (b) upon the specific action and intensity of the thymic influence.     

Functional effects of thymopoietin32-36 (TP5) on cytotoxic lymphocyte precursor units (CLP-U). I. Enhancement of splenic CLP-U in vitro and in vivo after suboptimal antigenic stimulation

Cytotoxic lymphocyte precursor units (CLP-U) were enumerated in the spleens of C57BL/6 mice 3 days after i.p. injections of synthetic thymopoietin32-36 (TP5). One hundred to 1000 ng TP5/mouse potentiated splenic CLP-U, this effect being detectable only after suboptimal allogeneic sensitization (with 1.2 x 10(5) mitomycin-C treated DBA cells). This elevation of CLP-U persisted in the injected mice for at least 14 days. Control peptide did not affect CLP-U. In vitro incubation of 0.01 to 0.1 ng/ml of TP5 with normal C57BL/6 spleen cells also enhanced CLP-U after suboptimal allogeneic stimulation; high concentrations of TP5 caused suppression of CLP-U and this was detectable with optimal sensitization conditions. Thus TP5, in vitro and in vivo, appears to regulate immune responsiveness and this regulation varies with TP5 dosage and with the immune stimulus.     

Effect of temperature on morphology and DNA-content of polytene chromosomes in Drosophila

Feulgen-DNA contents and chromosome lengths and projected areas were measured in salivary gland nuclei from Drosophila prepupae which had developed at 25° or 15° C. Nuclei from a given prepupa fell into 3 to 5 DNA classes corresponding to different levels of polyteny. The 15° nuclei tended to fall into higher classes than those from 25°-reared animals, and their chromosomes were, on average, about 50% wider. Chromosomes within a given DNA class did not differ significantly in mean area, length or width between the temperature groups, and slight apparent differences in mean DNA content were attributable to systematic microdensitometric errors associated with differences in the spreading behaviour of the nuclei. On cytological examination, chromosomes from the two temperature groups differed mainly in width and stain intensity, but some other differences in appearance could not be accounted for by levels of polyteny. The mean length of the chromosome complement was about 400 m. From one polytenic level to the next the chromosomes increased by about 10% in length, 40% in width and 17% in mean absorbance. The DNA content approximately doubled; small apparent deviations from the 12 ratio could have been due to microdensitometric error or to underreplication of heterochromatin.