Immobilization of alcohol dehydrogenase by gel entrapment of cells of Saccharomyces cerevisiae

A stable immobilized preparation of alcohol dehydrogenase (ADH) (EC 1.1.1.1) was obtained by entrapment of ADH-containing Saccharomyces cerevisiae cells in polyacrylamide, polymerized by gamma-rays (100 kR). The permeability barrier for the substrate through the cell membrane was found to be eliminated on entrapment. The stability characteristics, pH-activity profile and other properties of the entrapped ADH are presented. A four-fold enhancement in K_m for NAD⁺ was observed on entrapment, whereas K_m for ethanol was not altered.     

Pulmonary formation of prostacyclin in man

The pulmonary formation of prostacyclin (PGI₂), as reflected by the difference in concentration of pulmonary and systematic arterial radioimmunoassayed 6-keto-PGF_1α, was determined in six healthy waking subjects. The systematic arterial 6-keto-PGF_1α levels were low (50 pg/ml), and no evidence of pulmonary formation and release of the compound was noted. In other experiments systemic arterial 6-keto-PGF_1α levels were determined in patients prior to and during artificial ventilation, as well as during and after occlusion of the pulmonary circulation (extra-corporeal circulation, ECC). The arterial 6-keto-PGF_1α concentration prior to artificial ventillation was 17±4 pg/ml, i.e. within the range observed in the healthy subjects. During artificial ventilation the arterial levels of 6-keto-PGF_1α increased to 191±21 pg/ml, suggesting that pulmonary formation of PGI₂ was stimulated. In the patients subjected to ECC with occluded pulmonary circulation the arterial content of 6-keto-PGF_1α was stabilised at an elevated level (120−170 pg/ml). Following re-establishment of the pulmonary circulation the arterial concentrations of 6-keto-PGF_1α increased markedly, to 284±50 pg/ml. It is suggested that the basal pulmonary formation of PGI₂ in man is low or non-existent, and that enhanced formation of the compound in the lungs is a consequence of intervention with normal pulmonary ventilation or perfusion.     

Effects of β-endorphin on brain serotonin metabolism

Human β-endorphin (15 μg) administered intracisternally increased concentrations of serotonin (5HT) and its metabolite, 5-hydroxyindoleacetic. acid (5-HIAA), in brain stem and hypothalamus and decreased 5-HIAA concentrations in hippocampus. These data are compatible with the hypothesis that β-endorphin increases 5HT turnover in brain stem and hypothalamus and decreases 5HT turnover in hippocampus. β-endorphin increased in brain stem and hypothalamus and decreased in hippocampus the rate of pargyline-induced decline of 5-HIAA. β-endorphin decreased the rate of pargyline-induced accumulation of 5HT in all these brain regions. The probenecid-induced accumulation of 5-HIAA in brain stem was decreased by β-endorphin. These data are compatible with the hypothesis that β-endorphin increases release of 5HT from neurons in brain stem and hypothalamus and decreases release of 5HT from neurons in hippocampus. The data require further a hypothesis that β-endorphin either decreases 5HT reuptake in these three brain regions or increases 5-HIAA egress from brain.     

Secondary flow in the human common carotid artery imaged by MR angiography.

The blood flow in arteries affects both the biology of the vessels and the development of atherosclerosis. The flow is three-dimensional, unsteady, and difficult to measure or to model computationally. We have used phase-shift-based magnetic resonance angiography to image and measure the flow in the common carotid arteries of a healthy human subject. There was curvature of the vessels and thin-slice dynamic flow imaging showed evidence of the presence of secondary motions. Flexing the cervical spine straightened the vessels and reduced the asymmetry of the flow.     

Reversible inhibition by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid of the plasma membrane (Ca(2+)+Mg2+)ATPase from kidney proximal tubules

Calcium accumulation by purified vesicles derived from basolateral membranes of kidney proximal tubules was reversibly inhibited by micromolar concentrations of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), an inhibitor of anion transport. The inhibitory effect of this compound on Ca2+ uptake cannot be attributed solely to the inhibition of anion transport: (Ca(2+)+Mg2+)ATPase and ATP-dependent Ca2+ transport, respectively. The rate constant of EGTA-induced Ca2+ efflux from preloaded vesicles was not affected by DIDS, indicating that this compound does not increase the permeability of the membrane vesicles to Ca2+. In the presence of DIDS, the effects of the physiological ligands Ca2+, Mg2+, and ATP on (Ca(2+)+Mg2+)ATPase activity were modified. The Ca2+ concentration that inhibited (Ca(2+)+Mg2+)ATPase activity in the low-affinity range decreased from 91 to 40 microM, but DIDS had no effect on the Km for Ca2+ in the high-affinity, stimulatory range. Free Mg2+ activated (Ca(2+)+Mg2+)ATPase activity at a low Ca2+ concentration, and DIDS impaired this stimulation in a noncompetitive fashion. The inhibition by DIDS was eliminated when the free ATP concentration of the medium was raised from 0.3 to 8 mM, possibly due to an increase in the turnover of the enzyme caused by free ATP accelerating the E2----E1 transition, and leading to a decrease in the proportion of E2 forms under steady-state conditions. Alkaline pH totally abolished the inhibition of the (Ca(2+)+Mg2+)ATPase activity by DIDS, with a half-maximal effect at pH 8.3.(ABSTRACT TRUNCATED AT 250 WORDS)     

C-methyl phenolics from Qualea species

The trunk wood of Qualea labouriauana contains, besides (2R)-5,7,4′-trihydroxy-3′-methoxy-6,8-dimethylflavanone, (2R)-5,7,4′-trihydroxy-8-methylflavanone, the biosynthetically interesting 2,2′-dihydroxy-4,6,4′,6′-tetramethoxy-3,3′-dimethylbenzophenone. From the trunk wood extract of Q. paraensis the first named flavanone crystallized out directly.     

Effect of restrictive temperature on cell wall synthesis in a temperature-sensitive mutant of Bacillus stearothermophilus.

A temperature-sensitive mutant of Bacillus stearothermophilus, TS-13, was unable to grow above 58 degrees C, compared to 72 degrees C for the wild type. Actively growing TS-13 cells lysed within 2 h when exposed to a restrictive temperature of 65 degrees C. Peptidoglycan synthesis stopped within 10 to 15 min postshift before a shut down of other macromolecular syntheses. Composition of preexisting peptidoglycan was not altered, nor was new peptidoglycan of aberrant composition formed. No significant difference in autolysin activity was observed between the mutant and the wild type at 65 degrees C. Protoplasts of TS-13 cells were able to synthesize cell wall material at 52 degress C, but not at 65 degrees C. This wall material remained closely associated with the cell membrane at the outer surface of the protoplasts, forming small, globular, membrane-bound structures which could be visualized by electron microscopy. These structures reacted with fluorescent antibody prepared against purified cell walls. Production of this membrane-associated wall material could be blocked by bacitracin, which inhibited cell wall synthesis at the level of transport through the membrane. The data were in agreement with previous studies showing that at the restrictive temperature this mutant is unable to alter its membrane fatty acid and phospholipid composition with temperature such that it is not able to maintain a membrane lipid composition which permits normal membrane function at the restrictive temperature.     

Isolation and characterization of a trypsin inhibitor fromVigna sinensis seeds

The major trypsin inhibitor ofVigna sinensis cv. seridó was isolated and shown to be devoid of chymotrypsin inhibiting activity. It has a molecular weight of 9800 as determined by gel electrophoresis and a pI of 5.0. The activity of the inhibitor was decreased by treatment with trinitrobenzenesulfonic acid, suggesting that it isa LYS-X type trypsin inhibitor. Selfassociation of the molecule was demonstrated both in 1% sodium dodecylsulfate and inacidic (pH 2.4) conditions.     

Mutations of Chinese Hamster Somatic Cells from 2-Deoxygalactose Sensitivity to Resistance

In Chinese hamster somatic cells, the spontaneous change of phenotype from 2-deoxygalactose sensitivity to resistance was studied using fluctuation test experiments à la Luria and Delbrück (1943) for four Chinese hamster cell strains derived from V79. The results are consistent with true mutational events. The mutation rates are in the range of 1 to 3.5 X 10(-5) per cell per generation. The relationship between the 2-deoxyglactose resistance and the galactokinase markers is discussed.     

Cytochemistry and freeze-fracture of membranes isolated from the electrocyte of Electrophorus electricus (L.)

Summary Membranes were isolated from the main electric organ of Electrophorus electricus and studied by means of cytochemistry and freezefracture. The membrane fractions consisted of vesicles inside-in as determined by localization of anionic sites using colloidal iron and cationized ferritin particles. The anionic sites were not homogeneously distributed on the surface of the vesicle. Freeze-fracture showed the presence of intramembranous particles associated with either protoplasmic (P) or extracellular (E) faces of the membrane. Regions of the membrane without particles were observed. The results are discussed in relation to the existence of association between intramembranous particles and membrane receptors.For all correspondence