Use of Erythrocytes Sensitized with Purified Pneumococcal Polysaccharides for the Assay of Antibody and Antibody-producing Cells

A method was described for the sensitization of erythrocytes with purified type-specific pneumococcal polysaccharide antigens using chromium chloride as a coupling agent. Erythrocytes so sensitized can be used in routine passive hemagglutination and hemolysis tests as well as in the technique of localized hemolysis-in-gel for the detection of specific antibody and specific antibody-producing cells, respectively.     

Mechanism of Antifungal Action of 2,4,5,6-Tetrachloroisophthalonitrile

Studies of the toxicity of 2,4,5,6-tetrachloroisophthalonitrile (TCIN) to cells of Saccharomyces pastorianus and Neurospora crassa showed that 2 to 4 μg/ml of the toxicant were required to inhibit growth. Several thiol compounds reversed toxicity to growth. Glucose oxidation was severely impaired in treated cells of both test organisms. The toxicant at 2 or 4 μg/ml markedly reduced soluble thiol content of these cells. Bound thiol content was less affected in S. pastorianus cells than soluble thiol content. Uptake of toxicant by yeast cells was accompanied by formation of derivatives, some of which resembled those formed by reaction of glutathione with TCIN in vitro. Coenzyme A, glutathione, and 2-mercaptoethanol readily formed derivatives with TCIN in vitro. The nature of these derivatives was studied using 2-mercaptoethanol products as model substituents. Four derivatives were formed with 2-mercaptoethanol each with similar functional groups but showing dissimilar degrees of mobility during silica gel chromatography. Evidence indicated that these are derivatives of TCIN in which 1 to 4 of the halogens has been substituted by 2-mercaptoethanol. The mechanism of fungicidal action of TCIN is attributed to thiol inactivation.     

Partial Purification and Characterization of the Vacuolar H+-ATPase of Mammalian Synaptic Vesicles

Several major proteins of synaptic vesicles from rat or cow brain sediment as a large complex on sucrose density gradients when solubilized in nonionic detergents. A vacuolar H(+)-ATPase identified by sensitivity to bafilomycin A1 appears to be associated with this oligomeric protein complex. Two subunits of this complex, synaptic vesicle proteins S and U, correspond to the 57-kDa (B) and 39-kDa accessory (Ac39) subunits, respectively, of bovine chromaffin granule vacuolar H(+)-ATPase as shown by Western immunoblot analysis. The five subunits of the oligomeric complex constitute approximately 20% of the total protein of rat brain synaptic vesicles. Taken together, these results strongly suggest that the abundant, multisubunit complex partially purified from brain synaptic vesicles by density gradient centrifugation is a vacuolar H(+)-ATPase. Bafilomycin A1 completely blocks proton pumping in rat brain synaptic vesicles as measured by [14C]methylamine uptake and also blocks catecholamine accumulation measured by [3H]dopamine uptake. Moreover, ATPase activity, [14C]methylamine uptake, and [3H]dopamine uptake are inhibited by bafilomycin A1 at similar I50 values of approximately 1.7 nmol/mg of protein. These findings indicate that the vacuolar H(+)-ATPase is essential for proton pumping as well as catecholamine uptake by mammalian synaptic vesicles.     

Chromosomal localization of ARSB,the gene for human N-acetylgalactosamine-4-sulphatase

Summary A deficiency of N-acetylgalactosamine-4-sulphatase (G4S, gene symbol ARSB), results in the accumulation of undegraded substrate and the lysosomal storage disorder, Maroteaux-Lamy syndrome (mucopolysaccharidosis type VI). In situ hybridization using an ³H-labelled human G4S genomic DNA fragment to human metaphase chromosomes localized ARSB to chromosome 5q13–5q14. This location is consistent with, an refines, previous chromosomal assignments based on the expression of human G4S in somatic cell hybrids.     

Ocean current variability and the spawning season of Hawaiian reef fishes

Synopsis The spawning patterns of four Hawaiian reef fishes with similar reproductive habits, but different biogeographic distributions were studied from September 1980 to October 1981. Two species are Hawaiian endemics [Centropyge potteri (Pomacanthidae) and Chaetodon multicinctus (Chaetodontidae)] and the other two have pan-tropical Pacific distributions [Ctenochaetus strigosus and Zebrasoma flavescens (Acanthuridae)]. All showed increased spawning activity from January to July, the prevalent pattern among a majority of coastal marine fishes in Hawaii. Environmental correlatives to the period of peak reproduction include cycles of (a) daylight length and temperature which probably function as proximate cues and (b) seasonal variations in ocean current patterns which may ultimately affect survival of larvae and dispersal. Peak reproduction takes place during months when (1) mesoscale eddies most likely occur, and (2) the probability is greatest of drifting objects remaining near the islands.     

Spawning behavior ofChaetodon multicinctus (Chaetodontidae); pairs and intruders

Synopsis Spawning by the banded butterflyfish,Chaetodon multicinctus (Chaetodontidae) was observed on coral reefs off Kona, Hawaii. These fish occurred in male-female pairs during normal daytime activities, a behavior which is typical for the family. Courtship is also a paired male-female activity. During spawning, however, other individuals (males?) may intrude on the spawning pair. Spawning typically takes place at least a meter or two above the bottom. The spawning position consists of the male below and behind the female with his snout against the female's ventral flank or anal fin area. Intruding individuals may join in when the pair is in position and about to spawn. Intruders line-up against the male in the same position as he is against his female. Underwater photographs are included to illustrate these behaviors.     

Developmental Changes in the Calcium Dependency of γ-Aminobutyric Acid Release from Isolated Growth Cones: Correlation with Growth Cone Morphology

We have investigated the development of Ca2+-dependent gamma-[3H]aminobutyric acid [( 3H]GABA) release in superfused growth cone fractions isolated from rats between the postnatal ages of 1 and 11 days. We have compared this release with the overall morphology of the subcellular fractions, and identified those structures taking up [3H]GABA by electron microscopical autoradiography. In fractions isolated from rats between 1 and 5 days, K+-evoked [3H]GABA release was completely independent of extracellular Ca2+. After 5 days a Ca2+ dependency appeared, which increased with age, such that by 10 days approximately 50% of the K+-evoked release was Ca2+ dependent. Electron microscopical analysis showed that, at all ages, large numbers of GABAergic growth cones were present in the subcellular fractions. Up to postnatal day 5, the growth cones were synaptic vesicle sparse but, after this age, increasing numbers of synaptic vesicle-containing growth cones were seen. These results suggest that during maturation of GABAergic growth cones into synapses there is, initially, a mechanism for release that is independent of extracellular Ca2+ and that the appearance of a Ca2+-dependent [3H]GABA release from growth cones correlates with the appearance of synaptic vesicles.     

Aberrant mitochondrial processing of chimaeric import precursors containing subunits 8 and 9 of yeast mitochondrial ATP synthase

A set of chimaeric precursors which contain the same leader sequences but different passenger proteins has been analyzed for the site of protease cleavage following import into yeast mitochondria. Each precursor comprises the leader of Neurospora crassa subunit 9 of mitochondrial ATP synthase fused to subunit 8 or 9 of the corresponding yeast enzyme. Precursors containing the first five residues of mature N. crassa subunit 9 interposed between the leader and the yeast passenger protein were cleaved at the natural site of the N. crassa subunit 9 precursor. Direct fusions without interposed sequences were cleaved at novel sites. Cleavage occurred between the 3rd and 4th residues of yeast subunit 8, but for yeast subunit 9, cleavage occurred within the leader, 8 residues upstream of the passenger protein.