Nitrogen fixation (C2H2 reduction) in soil samples from rhizosphere of rice grown under alternate flooded and nonflooded conditions

Summary In a greenhouse study the influence of alternate flooded and nonflooded conditions on the N₂-ase activity of rice rhizosphere soil was investigated by C₂H₂ reduction assay. The soil fraction attached to roots represent the rhizosphere soil. Soil submergence always accelerated N₂-ase and this effect was more pronounced in planted system. Moreover, rice plant exhibited phase-dependent N₂-ase with a maximum activity at 60 days after transplanting. The alternate flooded and nonflooded regimes resulted in alterations of the N₂-ase activity. Thus, the N₂-ase activity increased following a shift from nonflooded to flooded conditions, but the activity decreased when the flooded soil was returned to nonflooded condition by draining. However, the differential influence of the water regime on N₂-ase was not marked in prolonged flooded-nonflooded cycles. Microbial analysis indicated the stimulation of different groups of free-living and associative N₂-fixing microorganisms depending on the water regime.     

Molecular species of epidermal growth factor carrying immunosuppressive activity

The suppression of antibody formation to sheep red cells in mice by partially purified fractions of mouse submaxillary gland was shown to be caused by epidermal growth factor (EGF). Purification of EGF by the method of Savage and Cohen resolved three components referred to as EGF a, EGF b, and EGF c. All three induced premature eye opening in neonatal mice, but only EGF a (identified as EGF 1-53) had full immunosuppressive activity. EGF c was shown by micropeptide mapping of chymotryptic and thermolytic digests and amino-terminal analysis to differ from EGF a only by the presence of beta-aspartyl instead of an asparaginyl residue. EGF b differed from EGF a in that it lacked the N-terminal asparagine. EGF shortened enzymatically at its carboxy terminal by two or five amino acids did not have any immunosuppressive activity. These findings suggest that, in contrast to some other biological effects of EGF, intact amino and carboxy terminals are required for the expression of immunosuppressive activity.     

Myxobacterial slime and proteolytic activity

An extracellular protein-polysaccharide-lipide (PPL) complex from exponentially growing cultures of Myxococcus virescens was purified by phosphate precipitation and gel chromatography. The high molecular weight slime polymer appeared homogenous upon isoelectric focusing. The PPL complex exhibited proteolytic activity against gelatin and the activity was only partly reduced by heat treatment. The function of the slime polymer as protein denatured was studied. The complex formed micelles similar to anionic detergents and it inhibited the precipitation and coagulation of proteins by trichloroacetic acid. Lysozyme was totally inactivated when treated with the PPL complex. By gel chromatography binding studies, the PPl complex was found to bind lysozyme in the ratio of 1 to 5.8 (w/w). After separation of added protein from the complex the anticoagulation effect on the protein remained. The biological function of the PPL complex was demonstrated with hemoglobin. When all susceptible peptide bonds in PPL-treated hemoglobin were hydrolyzed by trypsin only 20% in the urea-denatured protein were attacked. The combined role of slime and proteolytic activity is discussed.Abbreviations Used PPL protein-polysaccharide-lipide - TCA trichloroacetic acid - BSA bovine serum albumin - Tris tris-(hydroxymethyl)aminomethane - CMC critical micelle concentration - DNFB 2,4-dinitrofluorobenzene - DNP N-dinitrophenyl - SDS sodium dodecylsulphate - H.U. Hultin units     

Flavone 5-O-glucosides from Daphne sericea

The aerial parts of Daphne sericea yielded two new flavonoids, luteolin 7-methyl ether 5-β-d-glucoside and luteolin 7,3′-dimethyl ether 5-β-d-glucoside, as well as luteolin 7-methyl ether, isovitexin, apigenin and its 7-β-d-glucoside.     

Studies on antiviral glycosides. II. Mode of action for virucidal effects on Sendai virus

Treatment of Sendai virus with $\text{p-(sec-}\text{butyl)-phenyl-6-chloro-6-deoxy-β-}\text{d}\text{-glucopyranoside}$, followed by freezing and thawing resulted in a loss of hemolytic and cell fusion activities as well as infectivity without affecting hemagglutinating and neuraminidase activities. The anti-hemolytic activity of this compound was reversed by the addition of phosphatidyl choline to the virus samples. $\text{p-}\text{Azidophenyl-6-chloro-6-deoxy-β-}\text{d}\text{-[}{}^3\text{H]glucopyranoside}$ was successfully used for photoaffinity labeling of a specific virion site, and we confirmed the affected site of the glucoside to be the lipid components in the viral envelopes.     

Incorporation of3H-thymidine into DNA and the activity of alkaline phosphatase in zinc-deficient fetal rat brains

The incorporation of³H-thymidine into DNA in the brains of the 17-day and 20-day old rat fetuses was significantly reduced by maternal zinc restriction during pregnancy. The activity of the enzyme thymidine kinase (EC 2.7.1.21) was similarly reduced in the zine-deprived fetal brains on days 14 and 20 of gestation, but not on day 17. Fetal brain alkaline phosphatase (EC 3.1.3.1) was significantly depressed by maternal zinc deprivation on days 17 and 20 of pregnancy. The data suggest an association between thymidine kinase and the reduced incorporation of³H-thymidine into DNA in the brains of 20-day old fetuses but not in animals on day 17. Alkaline phosphatase was however depressed at this stage. The suggestion is made that because of the complexity of brain development, future biochemical studies in this area should concern specific structures in the brain at particular critical stages during neurogenesis.     

Antimicrobial defense mechanisms in the horseshoe crab, Limulus polyphemus: preliminary observations with heat-derived extracts of Limulus amoebocyte lysate.

Heat-derived (60°C) extracts of Limulus amoebocyte lysate (LAL) were found to contain potent “broad-spectrum” antimicrobial activity. Additional heating of the LAL extracts to 100°C for 30 min completely inactivated the antimicrobial activity and served as a control. Antimicrobial activity was observed over a temperature range of 0° to 37°C (higher temperatures not tested) with greatest activity at 37°C. Antimicrobial activity of LAL extracts was variable when tested against Gram-negative bacteria of the family Enterobacteriaceae. A twofold concentration of the extracts resulted in a significant decrease in antimicrobial effectiveness. Dialysis of single- and double-strength LAL extracts against deionized water produced a marked and significant enhancement of antimicrobial activity against both resistant and sensitive species, confirming the presence of a dialyzable inhibitor(s). Dialyzed LAL extracts were active against 13 of 14 species of Enterobacteriaceae tested. Two strains of Pseudomonas aeruginosa were susceptible as were two of three Gram-positive cocci tested. Highly sensitive bacterial species were rapidly killed with a greater than 90% reduction in viable counts occurring within the first 30 min of reaction time. Dialyzed LAL extracts also possessed considerable antifungal activity. The role of the Limulus polyphemus amoebocyte in defense against microbial invasion and dissemination is discussed.     

A new biologically active phenolic from Cattleya trianaei

A new phenolic, hydroxyeucomic acid, and dopamine were isolated from Cattleya trianaei and their biological activities examined.