Ribosomal dysentery vaccine. III. An immunochemical and serological study]

Sh. sonnei rib oscmes, isolated by differential centrifugation, were previously shown to be highly protect ive against experimental keratoconjunctivitis in guinea pigs. Immunochemical study showed that ribosomal preparations were not uniform in their antigenic composition: as a result of immunoelectrophoretic analysis with the use of anti-ribosomal hyperimmune rabbit antisera, these preparations were found to contain up to 4 antigenic components with different migration rate. The anodic component with the highest elections obtained by the method of Boivin and Grasset and could be inactivated at 60 degrees C or by treatment with trypsin or RNA-se, which suggested its ribonucleoprotein nature. The second thermolabile antigenic component was found to have a moderate anodic mobility and, judging by the results of enzymatic treatment, seemed to be protein. Other antigens with low mobility were resistant to trypsin and RNA-se; one of them, forming a weak precipitation line, could be identified as endotoxin by its antigenic specificity. The use of tanned and ribosome-coated erythrocytes allowed to determine the level of antiribosomal antibodies in the passive hemagglutination test and to evaluate the serological activity of ribosomal preparations in the hemagglutination inhibition test (the minimum inhibiting concentration of ribosomes was 1--2 microgram/ml). The specificity of serological reactions was mainly determined by a highly mobile nucleoproteid component.     

Ribosomal dysentery vaccine. II. Biological trials of active protection for guinea pigs and mice]

Ribosomal vaccine from Sh. sonnei injected subcutaneously once or twice in physiological saline or in Freund's complete adjuvant produces a marked protective effect against experimental keratoconjunctivitis in guinea pigs. Inhibition of the protective effect by high doses (above 100 microgram) of ribosomal vaccine is evident after a single, but not repeated injections. Protective effect in mice is achieved by immunization with very low doses of ribosomal vaccine: ED50 is 1.2 ng after challenge with 5.6 LD50. The nature of immunogenic factor responsible for the biological activity of the ribosome vaccine is still obscure. In contrast to Boivin's antigen, ribosomal preparations, even in high doses (1000--2000 microgram), have no toxic effect on mice and guinea pigs.     

Ribosomal dysentery vaccine. Study of response and antigenic activity in healthy volunteers

In earlier studies Shigella sonnei ribosomal vaccine was shown to be highly protective for guinea pigs and monkeys. The object of the present study, carried out in 20 healthy volunteers, was the safety and the antigenic activity of this vaccine. The subcutaneous injection of the ribosomal vaccine in doses of 100 micrograms and 200 micrograms produced no febrile reactions nor biochemical and histological changes. The minimal local reaction was observed after injection into the subscapular region: in this case 200 micrograms of the vaccine produced neither severe, nor moderate reactions. A single injection of this dose led to a more than 4-fold rise in the levels of total and cysteine-resistant O-antibodies, as well as to the prolonged elevation of the complement level in the serum.     

Isolation and physical chemical properties of rat hemopexin]

Rat hemopexin was purified by a procedure involving three different steps : ammonium sulfate precipitation, rivanol precipitation and DEAE-cellulose chromatography with concave gradient of molarity. Purity of the preparation was checked by three different methods : analytical ultracentrifugation, immunoelectrophoresis and acrylamide gel electrophoresis. The principal physical properties were studied. The amino acid and carbohydrate composition was determined and compared with that of human and rabbit hemopexin.     

Isolation, chemical, and physical properties of alpha-1-antitrypsin.

A method of isolation of alpha-1-antitrypsin (alpha-1-AT) in good yield from normal human plasma is described. A key step was affinity chromatography employing an antiserum which had been depleted of alpha-1-AT antibodies. The final preparations were homogeneous by immunological and physicochemical criteria. The specific activity of the purified alpha-1-AT was 0.363 mg of active bovine trypsin inhibited per 1.0 mg of inhibitor. Polyacrylamide gel patterns at both alkaline and acid pH of highly pure preparations frequently, but not invariably, showed multiple hands. Molecular weight studies by sedimentation equilibrium ultracentrifugation in aqueous buffer and in 6 M guanidine as well as sodium dodecyl sulfate polyacrylamide gel electrophoresis suggest that alpha-1-AT is a single polypeptide chain having a molecular weight of 49,500. Other physical and chemical properties of the inhibitor are described. A limited N-terminal sequence (Glu-Asp-Pro-Gln-Gly-Asx-Ala-Ala) was obtained. It was found that alpha-1-AT easily forms polymers and higher aggregates when exposed to denaturing agents such as 8 M urea and 6 M guanidine. The results suggest that aggregation is determined by both covalent and noncovalent forces.     

Rabbit liver transglutaminase: physical, chemical, and catalytic properties.

Transglutaminase (R-glutaminyl-peptide:amine alpha-glutamyl-yltransferase [EC 2.3.2.13]) has been purified to apparent homogeneity from extracts of rabbit liver. The enzyme is a single polypeptide chain of approximately 80 000 molecular weight containing one catalytic site per molecule. That the isolated enzyme is the rabbit counterpart of the well-characterized guinea pig liver transglutaminase is evidenced by the similarities in their amino acid compositions and in their enzymic activities toward several substrates, together with the fact that the isolated rabbit enzyme is immunologically distinct from both rabbit plasma and rabbit platelet blood coagulation factor XIII. A striking difference between the catalytic activities of the rabbit and guinea pig enzymes is the low activity of rabbit transglutaminase for hydroxylamine incorporation into benzyloxycarbonyl-L-glutaminylglycine, a reaction for which the guinea pig enzyme shows a high reactivity. This finding reveals the cause of error in an earlier report (Tyler, H.M., and Laki, K. (1967) Biochemistry 6, 3259) that rabbit liver contains little, if any, of the enzyme. Preparation of, and analytical data on, several glutamine-containing peptide derivatives used in this study are reported here.     

Ornithine transcarbamylase of rat liver. Kinetic, physical, and chemical properties.

Ornithine transcarbamylase of rat liver has been purified to homogeneity. The purified enzyme of specific activity 870 to 920 focuses as a single protein at pH 7.2. At pH 7.7, the Km for carbamyl phosphate is 0.026 mM, and the Km for ornithine is 0.04 mM. The inhibition constants of a number of amino acids that act as competitive inhibitors of the enzyme are reported. The native enzyme of Mr = 112,000 is composed of three subunits of Mr = 39,600 +/- 1,000. Chemical evidence indicates that the subunits are identical in amino acid composition and amino acid sequence. The amino acid sequence of the NH2-terminal region of ornithine transcarbamylase is Ser-Gln-Val-Gln-Leu-Lys-Gly-Ser-Asp-Leu-Leu-Thr-Leu-Lys-Asn-(Phe)-X-Thr-X-Glu-Ile-Gln-Tyr-Met-.     

Relationships between cyanobacterial production and the physical and chemical properties of a Midwestern Reservoir,USA

Drinking water reservoirs in agricultural dominated watersheds are particularly vulnerable to cyanobacterial blooms. A major byproduct of cyanobacteria is the production of objectionable taste and odor compounds such as geosmin. During May 1997 to September 1998, we studied spatial and temporal patterns of cyanobacterial abundance and composition with respect to a series of physical and chemical properties in Clinton Lake, located in east central Kansas, USA. Our results suggest that nutrients (in particular TN, NO₃–N concentrations), turbidity, and hydrologic regime all played potentially important roles in regulating cyanobacterial production. Specifically, low levels of nitrogen coupled with the internal release of phosphorus from the lake sediment under brief periods of anoxia may have helped promote cyanobacterial blooms. There was also a strong association between cyanobacterial blooms, geosmin production, and most taste and odor events in Clinton Lake. Anabaena circinalis appeared to be the source for geosmin production as a result of senescing algal cells just after the primary die-off of cyanobacteria.     

Mung bean nuclease I. Physical, chemical, and catalytic properties.

A simplified purification procedure for mung bean nuclease has been developed yielding a stable enzyme that is homogeneous in regards to shape and size. The nuclease is a glycoprotein consisting of 29% carbohydrate by weight. It has a molecular weight of 39 000 as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The enzyme contains 1 sulfhydryl group and 3 disulfide bonds per molecule. It has a high content (12.6 mol %) of aromatic residues. Approximately 70% of the enzyme molecules contain a peptide bond cleavage at a single region in the protein. The two polypeptides, 25 000 and 15 000 daltons, are covalently linked by a disulfide bond(s). Both the cleaved and intact forms of the enzyme are equally active in the hydrolysis of the phosphate ester linkages in either DNA, RNA, or adenosine 3'-monophophate. The enzymatic activity of mung bean nuclease can be stabilized at pH 5 in the presence of 0.1 mM zinc acetate, 1.0 mM cysteine, and 0.001% Triton X-100. The enzyme can be inactivated and reactivated by the removal and readdition of Zn2+ or sulfhydryl compounds.     

Acid-activated carbons from almond shells: physical,chemical and adsorptive properties and estimated cost of production

A series of phosphoric-acid activated carbons were made from almond shells using six different activation or activation/oxidation methods. The carbons were compared to each other and to two commercial carbons in an effort to ascertain the relative value of the carbons in terms of yield, surface area, attrition, surface functional groups, organic uptake, metal uptake, as well as estimated cost of production. Of the six methods investigated, the method that produced the best overall performing almond shell carbon and least expensive carbon in terms of production cost was the “Air-Activation” method. This method involved the simultaneous activation and oxidation of almond shells under an air atmosphere.     

Patterns of temporal variation in Lake Titicaca. A high altitude tropical lake. I. Background,physical and chemical processes,and primary production

A statistical analysis is presented of patterns of variation in some physical, chemical, and biological variables for a 6 year series of data from the tropical, high altitude Lake Titicaca (Peru-Bolivia). ANOVA techniques and autocorrelation analyses were used to partition the variance in Titicaca, and in some comparison tropical and temperate series, into components with repeatable annual cycles and components attributable to other kinds of patterns.In Titicaca, insolation and stratification are highly seasonal in pattern of variation, although the amount of variance relative to means is small compared to temperate lakes. However, the seasonal pattern of physical variation is only weakly imposed on chemical and biological processes, to judge from analyses of silicate, oxygen, and primary production series. Comparable temperate series of primary production and chlorophyll a are much more seasonal.     

Chondronectin: physical and chemical properties

Chondronectin, the chondrocyte attachment factor, was purified from chicken serum and characterized as to its physical and chemical properties. From sedimentation equilibrium data it was found to have a native molecular weight of 175,800 +/- 800 and a subunit molecular weight of 55,540 +/- 800 in the presence of guanidinium chloride and cysteine, suggesting a trimeric structure linked by disulfide bonds. As visualized by electron microscopy after rotary shadowing, the protein appears compact and globular. The amino acid and carbohydrate compositions of chondronectin are distinct from fibronectin, the fibroblast attachment factor, and laminin, the epithelial cell attachment factor. The activity of chondronectin in promoting attachment of chondrocytes is stable to digestion by collagenase, elastase, and neuraminidase, but is destroyed by trypsin treatment. The data suggest that chondronectin is structurally and chemically distinct from fibronectin and laminin.