Surface haemagglutinating activity of Pseudomonas aeruginosa

Intact cells of several strains of Pseudomonas aeruginosa agglutinate papain-treated human erythrocytes. The agglutinating activity appears to reside in the surface layers of the bacterium-Pseudomonas surface haemagglutinin. This activity does not correlate with the existence of the internal PA-I and PA-II lectins, the presence of fimbriae or adherence to human buccal epithelial cells. Disruption of the bacterial cells by sonication abolishes their haemagglutinating activity. The intact cells of P. aeruginosa are also able to agglutinate rabbit, chicken, dog, guinea pig and sheep erythrocytes. This activity is generally higher with papain-treated erythrocytes, except those of rabbit in which lower haemagglutinating activity is observed after papain treatment. Optimal conditions for the haemagglutination are 37 degrees C and pH 6-7. Simple sugars do not inhibit, while fetuin and hydrophobic amino acids inhibit this activity. Exposure of the bacterial cells to proteolytic enzymes, EDTA or denaturating conditions abolish the haemagglutinating activity. These results indicate that the surface haemagglutinin is a protein which agglutinates red blood cells via hydrophobic interactions.     

Single Na+ channels activated by veratridine and batrachotoxin

Voltage-sensitive Na+ channels from rat skeletal muscle plasma membrane vesicles were inserted into planar lipid bilayers in the presence of either of the alkaloid toxins veratridine (VT) or batrachotoxin (BTX). Both of these toxins are known to cause persistent activation of Na+ channels. With BTX as the channel activator, single channels remain open nearly all the time. Channels activated with VT open and close on a time scale of 1-10 s. Increasing the VT concentration enhances the probability of channel opening, primarily by increasing the rate constant of opening. The kinetics and voltage dependence of channel block by 21-sulfo-11-alpha-hydroxysaxitoxin are identical for VT and BTX, as is the ionic selectivity sequence determined by bi-ionic reversal potential (Na+ approximately Li+ greater than K+ greater than Rb+ greater than Cs+). However, there are striking quantitative differences in open channel conduction for channels in the presence of the two activators. Under symmetrical solution conditions, the single channel conductance for Na+ is about twice as high with BTX as with VT. Furthermore, the symmetrical solution single channel conductances show a different selectivity for BTX (Na+ greater than Li+ greater than K+) than for VT (Na+ greater than K+ greater than Li+). Open channel current-voltage curves in symmetrical Na+ and Li+ are roughly linear, while those in symmetrical K+ are inwardly rectifying. Na+ currents are blocked asymmetrically by K+ with both BTX and VT, but the voltage dependence of K+ block is stronger with BTX than with VT. The results show that the alkaloid neurotoxins not only alter the gating process of the Na+ channel, but also affect the structure of the open channel. We further conclude that the rate-determining step for conduction by Na+ does not occur at the channel's "selectivity filter," where poorly permeating ions like K+ are excluded.     

Genomic and cDNA clones of the homeotic locus Antennapedia in Drosophila.

Homeotic genes are involved in the control of developmental pathways: dominant mutations at the Antennapedia locus of Drosophila, for example, lead to replacement of the antennae on the head of the fly by mesothoracic legs. Using a combination of chromosome walking and jumping, we have cloned a DNA region from Drosophila containing Antennapedia. Five DNA inversion rearrangements which are associated with the Antennapedia mutant phenotype were localized within a 25-kb region. Genomic DNA sequences from this area were used as hybridization probes to screen cDNA libraries prepared from Drosophila embryonic and pupal poly(A)+ RNA. A 2.2-kb cDNA sequence (903) was isolated which appears to derive from at least four non-contiguous chromosomal regions that span 100 kb. It includes the positions of the inversion breakpoints. A second cDNA of 2.9 kb (909) is composed of sequences from at least three chromosomal regions, two of which are similar or identical to sequences contained in the 903 clone but the third is derived from genomic DNA within a putative 903 intron. The unusual size and complexity of this locus are discussed.     

Efficiency of Glasshouse Pot Experiments Rotating versus Stationary Benches

Pot cultures of rice (Oryza sativa L., var. Colusa) in flooded soil were used to measure comparative uniformity of growth where mutual shading was the primary source of variation. Data were analyzed according to various divisions of each layout into blocks, sections, and replicate pots to compare statistical efficiencies.     

Release of Inorganic Phosphate from Irradiated Yeast: Radiation Biodosimetry and Evaluation of Radioprotective Compounds

When cells of bakers' yeast, Saccharomyces cerevisiae, were irradiated with ionizing radiation, inorganic phosphate, ninhydrin-reactive material, and substances absorbing at 260 mmu were released into the suspending medium. The amount of inorganic phosphate released depended on the radiation dose and on the temperature and pH during irradiation. The concentration of yeast cells did not affect the phosphate yield per milligram of yeast. It is suggested that the release of phosphate may serve as an index of the total radiation environment (i.e., as a biodosimeter) where radiation inactivation of microrganisms is of primary importance, e.g., in radiation preservation of foods. The somewhat limited range of the yeast biodosimeter (ca. 0.5 to 1.75 Mrad) may be extended by use of other more resistant microorganisms, such as bacterial spores. Compounds which have been reported as protecting microorganisms and mammals against the lethal effect of ionizing radiation also inhibited the radiation-induced release of inorganic phosphate from yeast. This phosphate release system is proposed as the basis for an economical, rapid supplement to screening procedures in the evaluation of radioprotective compounds.     

GENETICS OF PHYTOPATHOGENIC FUNGI. XI. A GENETIC STUDY OF AVIRULENCE DUE TO AUXOTROPHY IN PENICILLIUM EXPANSUM BY MEANS OF THE PARASEXUAL CYCLE

Nine color and 35 auxotrophic mutants were tested for their virulence, using the ‘Jonathan’ variety of apple; only mutants requiring methionine were avirulent. One tested arginine-requiring and the methionine-requiring mutants were avirulent for the ‘Red Delicious’ variety. In vitro and in vivo, supplementation studies indicated that the avirulence was probably related to the relatively low concentration of the required amino acids at the site of inoculation. One heterocaryon and the corresponding diploid, involving 2 avirulent methionine-requiring mutants, were virulent. Six mutant loci in 3 diploid strains could be assigned to 2 linkage groups by means of the parasexual cycle.     

Crystals of threonyl-tRNA synthetase from Thermus thermophilus. Preliminary crystallographic data

Crystals have been obtained of threonyl-tRNA synthetase from the extreme thermophile Thermus thermophilus using sodium formate as a precipitant. The crystals are very stable and diffract to at least 2.4 A. The crystals belong to space group P2(1)2(1)2(1) with cell parameters a = 61.4 A, b = 156.1 A, c = 177.3 A.