Surface Antigens of Smooth Brucellae

Surface antigens of smooth brucellae were extracted by ether-water, phenol-water, trichloroacetic acid, and saline and examined by immunoelectrophoresis and gel diffusion with antisera from infected and immunized rabbits. Ether-water extracts of Brucella melitensis contained a lipopolysaccharide protein component, which was specific for the surface of smooth brucellae and was correlated with the M agglutinogen of Wilson and Miles, a polysaccharide protein component devoid of lipid which was not restricted to the surface of smooth brucellae and was not correlated with the smooth agglutinogen (component 1), and several protein components which were associated with internal antigens of rough and smooth brucellae. Immunoelectrophoretic analysis of ether-water extracts of B. abortus revealed only two components, a lipopolysaccharide protein component, which was correlated with the A agglutinogen, and component 1. Component 1 from B. melitensis and B. abortus showed identity in gel diffusion tests, whereas component M from B. melitensis and component A from B. abortus showed partial identity with unabsorbed antisera and no cross-reactions with monospecific sera. Attempts to prepare monospecific sera directly by immunization of rabbits with cell walls or ether-water extracts were unsuccessful. Absorption of antisera with heavy fraction of ether-water extracts did not always result in monospecific sera. It was concluded (as has been described before) that the A and M antigens are present on a single antigenic complex, in different proportions depending upon the species and biotype, and that this component is a lipopolysaccharide protein complex of high molecular weight that diffuses poorly through agar gel. Components 1, A, and M were also demonstrated in trichloroacetic acid and phenol-water extracts. With all extracts, B. melitensis antigen showed greater diffusibility in agar than B. abortus antigens. After mild acid hydrolysis, B. abortus ether-water extract was able to diffuse more readily.     

The metabolism of 3,5-di-tert.-butyl-4-hydroxytoluene in the rat and in man

1. The major metabolites of 3,5-di-tert.-butyl-4-hydroxytoluene (BHT) in the rat are 3,5-di-tert.-butyl-4-hydroxybenzoic acid (BHT-acid), both free (9% of the dose) and as a glucuronide (15%), and S-(3,5-di-tert.-butyl-4-hydroxybenzyl)-N-acetylcysteine. 2. The mercapturic acid does not appear to derive from the usually accepted enzyme mechanism, and may involve a non-enzymic reaction between BHT free radical and cysteine. 3. The ester glucuronide and mercapturic acid found in rat urine are also the major metabolites in rat bile and must be responsible for the enterohepatic circulation. 4. Free BHT-acid is the main component in rat faeces. 5. In man, BHT-acid, free and conjugated, is a minor component in urine, and the mercapturic acid is virtually absent. The bulk of the radioactivity is excreted as the ether-insoluble glucuronide of a metabolite in which the ring methyl group and one tert.-butyl methyl group are oxidized to carboxyl groups, and a methyl group on the other tert.-butyl group is also oxidized, probably to an aldehyde group. 6. These differences in metabolism by the rat and by man are sufficient to account for the difference in excretion by the two species.     

Utilization of 5-Bromouracil by Thymineless Bacteria

Several thymineless Escherichia coli strains have been examined for their ability to replicate their deoxyribonucleic acid when bromouracil is substituted for thymine. The procedure we describe was used to identify a thymineless strain with characteristics relatively favorable to its use in bromouracil labeling experiments. In addition, mutants with an “absolute” thymine requirement could be easily distinguished from one with a “leaky” thymine requirement.     

THE STRUCTURE AND FUNCTION OF CENTRIOLES AND THEIR SATELLITES IN THE JELLYFISH PHIALIDIUM GREGARIUM

Testes of jellyfish Phialidium gregarium were fixed in 2 per cent OsO₄ in Veronal-acetate buffer at pH 7.4. Thin sections showed that in young spermatids the spindle fibers of the last maturation division are attached to satellites of the filament-forming centriole. In more mature spermatids this attachment is not observed. During the developmental phase, nine satellites can be observed emanating from the interspaces between the nine tubular triplets of this centriole. A circular region on each of the enlarged distal ends of the satellites attaches them to the cell membrane. The satellites apparently provide a firm anchor for the axial filament. Each of the epithelial cells covering the testis produces a single long flagellum. On the filament-forming centriole often a satellite can be observed to which tubules are attached. These tubules are 180 A in diameter and probably represent remnants of spindle fibers. It is suggested that the distal centriole has the ability to form several satellites or appendages at appropriate times during the cell cycle. These satellites are distinct from the daughter centrioles in that they are supportive structures: in certain phases of cell life, spindle fibers may attach to them, while in other instances the distal centriole and the flagellum it is forming are anchored by them.     

Golgi apparatus mediated polysaccharide secretion by outer root cap cells of Zea mays

Summary In the outer cap cells of roots of Zea mays, secretion is accompanied by hypertrophy of dictyosome cisternae with formation of large secretory vesicles. Vesicle contents are subsequently released from the protoplast by fusion of the vesicle membrane with the plasma membrane. The secreted material, a highly hydrated polysaccharide, was localized intracellularly by the periodic acid-Schiff reaction. Under appropriate conditions, the product moves outward through the cell wall after discharge from the protoplast, and appears as a droplet adhering to the root tip. Under conditions where the secretory product accumulates at the inner wall surfaces, no external droplet is formed.The secretory activity has an active phase that is sensitive to metabolic inhibitors and influenced by temperature (Q₁₀>2), and a passive phase that is independent of temperature, insensitive to metabolic inhibitors but sensitive to osmotic agents. The active phase is characterized by a temperature-independent periodicity (3 hours). Sucrose supplied to the growth medium increases the amount of polysaccharide secreted. Polysaccharide synthesis, segregation into vesicles, and discharge from the protoplast are assumed to require active metabolism; the step involving extrusion of polysaccharide through the cell wall region appears to be a passive process influenced by the degree of hydration of the polysaccharide and by cell turgor.Purdue University Agricultural Experiment Station Journal Paper No. 2967; Charles F. Kettering Research Laboratory Contribution No. 261.