Some Properties of Thiosulfate-Oxidizing Enzyme from Marine Heterotroph 16B

Thiosulfate-oxidizing enzyme has been demonstrated in cell-free extracts of the marine, thiosulfate-oxidizing pseudomonad strain 16B. The enzyme, partially purified by ion-exchange chromatography and calcium phosphate gel treatment, catalyzed the oxidation of thiosulfate to tetrathionate with the concomitant reduction of ferricyanide. Native but not mammalian cytochrome c was also reduced by the enzyme in the presence of thiosulfate. The enzyme was located exclusively in the supernatant of ultracentrifuged cell extracts. The most purified enzyme preparation, like intact cells, exhibited a temperature optimum of 30 to 31°C. However, it exhibited no definite pH optimum. At pH 6.1 to 6.3 and 30°C, the K_m for thiosulfate was 1.57 mM. At lower temperatures, the apparent K_m for thiosulfate increased, but the apparent maximum velocity remained virtually unchanged. Thiosulfate oxidation in intact cells exhibited an increase in the pH optimum at lower temperatures. The thiosulfate-oxidizing enzyme of marine heterotroph 16B is compared with thiosulfate-oxidizing enzymes from other bacteria, and the effect of temperature on the relationship between pH and thiosulfate oxidation is discussed with reference to the natural habitat of the bacterium.     

Disturbance and contrasting patterns of population structure in the brachiopod Terebratulina septentrionalis (Couthouy) from two subtidal habitats

The population structures of Terebratulina septentrionalis (Couthouy) from exposed upper rock surface and semi-cryptic rock wall habitats at 33 m depth in the Gulf of Maine differ. Over a 3-yr period, population densities were consistently higher in rock wall habitats. Although both populations were dominated by juveniles (1–4 mm shell length), size-frequency distributions constructed from upper rock surface and rock wall populations were significantly different, as a result of a greater frequency of large brachiopods (> 20 mm shell length) in rock wall populations. Prominent modes occurred at 14–15 mm shell length in upper surface populations and at 19–20 mm length in rock wall populations. Recruitment was higher in rock wall habitats where ambient light intensities were significantly lower than on upper rock surfaces. Differences in recruitment are either the result of larval selection for shaded rock walls or differential juvenile mortality between habitats. The larvae of Terebratulina settle on a diverse array of substrata. These include bedrock, sandy polychaete tubes and algae in upper surface habitats and bedrock, calcareous polychaete tubes, and ascidians in rock wall habitats. Individuals attached to polychaete tubes and algae in upper surface habitats do not attain large body size (> 13 mm shell length). It is suggested that these differences in population structure reflect the greater intensity of disturbance in upper surface habitats. For example, the cod, Gadus morhua (Linnaeus), ingests brachiopods attached to algae and polychaete tubes in this habitat. Gastropod predation affects brachiopods in upper surface habitats but not in rock wall habitats. Predation by gastropods and asteroids is not size-specific. These results are consistent with the hypothesis that predation contributed to the decline in the abundance and diversity of articulate brachiopods since the Mesozoic, and suggest that the restriction of recent populations to semi-cryptic rock wall and crevice habitats is, in part, controlled by disturbance.     

Differential recognition of the serologically defined HLA-A2 antigen by allogeneic cytotoxic T cells

A comprehensive analysis of human alloimmune cytotoxic T lymphocytes (CTLs) specific for the HLA-A2 antigen identified 11% of HLA-A2 positive cells as outliers. In total, 11 unrelated serologically indistinguishable, but distinguishable by cell-mediated lympholysis (CML) HLA-A2 positive outlier cells were identified. The outlier cells could be subdivided in two subgroups according to reactivity patterns obtained with CTLs directed against the HLA-A2 antigen of outlier cells and their inhibitory capacity in specific competitive inhibition experiments. Thus, the serologically defined HLA-A2 specificity can be divided into at least three subtypes using CTLs specific for the HLA-A2 antigen. Moreover, CTLs specific for an HLA-A2 subtype could be induced when responder cells expressed a different HLA-A2 subtype antigen. On the basis of several family studies, we conclude that the subtype HLA-A2 antigens are inherited in a codominant way.     

Analysis of human class I antigens by two-dimensional gel electrophoresis

Class I antigens were isolated by immunoprecipitation from cell extracts prepared from mitogenically stimulated and internally radiolabeled peripheral blood lymphocytes (PLBs). The precipitating antibodies used are monomorphic and recognize a determinant on the heavy chain of HLA-A, B, C antigens regardless of their allelic specificities when complexed with ₂m, or determinants on ₂m itself. Comparison of class I molecules isolated from 25 different homozygous typing cels (HTC) and analyzed by two-dimensional (2-D) gel electrophoresis allowed the identification of those HLA-A,13 locus specificities most common in the European Caucasoid population. Class I antigens isolated from HTC that are HLA identical are biochemically indistinguishable also. Evidence was obtained for the expression of additional class I antigens besides the HLA-A, B, C locus products: for some haplotypes, up to six class I genes may be active in mitogenically activated PBLs. No differences in molecular weight and isoelectric point of the class I heavy chains were observed between the antigens recognized by W6/32, the anti-heavy chain reagent, and anti- ₂m reagents. The nature of the mitogenic stimulus, i. e., pokeweed mitogen or phytohemagglutinin, was irrelevant with respect to the class I antigens isolated by this method. Using the HTCs as reference, a panel of HLA-B27 positive heterozygous cells was analyzed. Two types of HLA-B27 antigens, distinct by CML typing were represented. These two forms differed also in their biochemical properties. In addition, we obtained evidence for the existence of an A2 variant. This finding was likewise confirmed by CML typing.     

Growth rate stimulation of marine pseudomonads by thiosulfate

The oxidation of thiosulfate to tetrathionate exerts a definite growth rate stimulation in glucose, acetate, and yeast extract cultures of some marine pseudomonads. The failure to find this effect in earlier studies with terrestrial isolates may lie in the particular conditions used in the present experiments (constant pH, high ratio of thiosulfate to organic substrate) or in the different metabolic characteristics of the marine isolates.Contribution No. 3220 from the Woods Hole Oceanographic Institution.     

Activity of Microorganisms in Acid Mine Water I. Influence of Acid Water on Aerobic Heterotrophs of a Normal Stream

Comparison of microbial content of acid-contaminated and nonacid-contaminated streams from the same geographical area indicated that nonacid streams contained relatively low numbers of acid-tolerant heterotrophic microorganisms. The acid-tolerant aerobes survived when acid entered the stream and actually increased in number to about 2 × 10³ per ml until the pH approached 3.0. The organisms then represented the heterotrophic aerobic microflora of the streams comprised of a mixture of mine drainage and nonacid water. A stream which was entirely acid drainage did not have a similar microflora. Most gram-positive aerobic and anaerobic bacteria died out very rapidly in acidic water, and they comprised a very small percentage of the microbial population of the streams examined. Iron- and sulfur-oxidizing autotrophic bacteria were present wherever mine water entered a stream system. The sulfur-oxidizing bacteria predominated over iron oxidizers. Ecological data from the field were verified by laboratory experiments designed to simulate stream conditions.