Conformational studies of bacterial polysaccharides. II. Optical activity of some bacterial capsular polysaccharides.
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The conformation of xanthan has been investigated as a function of temperature, ionic strength, and polymer concentration. A reversible transition induced by temperature is demonstrated; the melting temperature (TM) is directly correlated to the total ionic-strength and is independent of the polymer concentration. Measurements of circular dichroism show that the polysaccharide exists in a combination of only two characteristic conformations (random and ordered), regardless of the temperature and the concentrations of salt and polymer. Hydrodynamic measurements show that the hydrodynamic volume of both conformations is almost constant over the range of temperature investigated. The mechanism proposed by Morris for melting is confirmed, and a multichain process is excluded. The birefringence stability of the concentrated solutions is discussed.
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Following immunization with bacteria (i.e. Salmonella johannesburg), rabbit spleen lymphocytes developed a specific blast response when the lymphocytes were stimulated with polysaccharide, the haptenic moiety of lipopolysaccharide. A clear cut dissociation was noted in the blast response induced by polysaccharide compared with those induced by lipopolysaccharide and lipid A. There was no correlation between the magnitude of the cellular responses and that of the antibody response. Moreover, there was less specificity at the cellular level than at the level of antibody secreted by cells. A decrease of 3H-thymidine incorporation was often observed after immunization, at the level of peripheral blood lymphocytes. An inhibitory effect of these cells was shown on the blast response of spleen lymphocytes with polysaccharide. A high blast response to Salmonella polysaccharide which could be observed in some non-immunized rabbits might be related to a natural sensitization of animals with the same or related unknown antigens which could not be recognized by anti-S. johannesburg antibodies.
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The influence of bacterial polysaccharide on the amount of the rosette-forming cells (RFC) in mice with decreased immunological reactivity (irradiation in a dose of 400 rad, gamma-rays, administration of cyclophosphamide in a dose of 200 mg/kg) was studied. Bacterial polysaccharide increased the amount of RFC in the intact, immunized and irradiated animals. However, polysaccharide produced no stimulating action on the RFC formation in mice treated with cyclophosphamide this confirming different nature of depression of the immunological reactivity under the effect of irradiation and cyclophosphamide.
The effect of typhoid bacterial polysaccharide on the primary and secondary immune response to SRBS was studied. The polysaccharide was shown to have both stimulating and depressive effect on the population of antibody-producing cells. This effect depended on the time and the number of polysaccharide injections. Thus, a single polysaccharide injection made on the day preceding immunization resulted in the maximum stimulation in the system of IgM- and IgG-producing cells, while the maximum depression of these cells could be observed after 2 polysaccharide injections: on the day preceding immunization and on the day of immunization. In the secondary immune response considerable stimulation of the populations of antibody-producing cells was observed after polysaccharide injections made on days 2 and 3 after reimmunization.
The effect of bacterial polysaccharide on the number of spleen cells producing antibodies against sheep's red cells was investigated in irradiated and unirradiated mice. The polysaccharide increased the number of antibody-producing cells both in lethally irradiated (psi rays) and in unirradiated mice, unimmunized with sheep's red cells. This increase was connected with the stimulation of proliferative processes in the recipient by the polysaccharide.
Agrobacterium sp. and related species which in the soil and in the rhizosphere of wheat accompany the fungus Gaemannomyces graminis var. tritici and cause take-all of the wheat roots produced polysaccharides in pure cultures (glucans, mannoglucans and galactomannoglucans). These polysaccharides were utilized better by the mycelium of G. graminis than glucose and polysaccharides of plant origin that occurred on the surface of wheat roots (the so-called mucigel). At lower concentrations these bacterial polysaccharides stimulated growth of wheat roots, higher concentrations (more than 0.1%) were inhibitory. Bacteria inoculated on the surface of wheat first inhibited and then stimulated the development of the plants and their growth. Changes in the growth rate of wheat, the rhizosphere of which was colonized by bacteria simultaneously with the fungus G. graminis and also some changes in the course of the disease of wheat roots caused by the fungus can be explained by the inhibitory or stimulatory effect of polysaccharides of accompanying bacteria.
A simple experiment is described for demonstrating the formation of a high molecular-weight polysaccharide from sucrose by a bacterial enzyme. The polysaccharide confers high viscosity on solutions. Dental students are encouraged to consider this as a component of the system leading to the formation of dental plaque and the initiation of carious lesions.
Phenol-water extracted rheumatoid synovial fluids and synovial fluid leukocytes contain an antigen immunologically identical to the Proprionibacterium group bacteria. The antigen was identified by counter-immunoelectrophoresis in 70% of rheumatoid synovial fluid leukocyte pellets and in 60% of rheumatoid synovial fluids. It was also present in 6% of nonrheumatoid fluids and in 22% of nonrheumatoid inflammatory fluid leukocytes. Antigen was not detectable in synovial samples before extraction. Synovial and bacterial antigens were further purified by proteolytic digestion and Sepharose 4B column chromatography. Biochemical and enzymatic studies of bacterial and synovial antigens were similar and consistent with a high molecular weight polysaccharide. Serum antibody to bacterial and synovial antigens was significantly less frequent in rheumatoid sera than in normal controls. The significance of demonstrating a bacterial polysaccharide primarily in rheumatoid synovial effusions is discussed.
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Immunofluorescence, quantitative immunoprecipitation, and inhibition of bacterial agglutination and passive hemagglutination indicate that cross-reactive antigenic determinants are present on the surface of Rhizobium trifolii and clover roots. These determinants are immunochemically unique to this Rhizobium-legume cross-inoculation group. The multivalent lectin trifoliin and antibody to the clover root antigenic determinants bind competitively to two acidic heteropolysaccharides isolated from capsular material of R. Trifolii 0403. The major polysaccharide is an antigen which lacks heptose, 2-keto-3-deoxyoctulosonic acid, and endotoxic lipid A. The minor polysaccharide in the capsular material of R. Trifolii 0403 contains the same antigen in addition to heptose, 2-keto-3-deoxyoctonate, and lipid A. The acidic polysaccharides of two strains of R. trifolii share the clover r-ot cross-reactive antigenic determinant despite other differences in their carbohydrate composition. Studies with monovalent antigen-binding fragments of anti-clover root antibody and Azotobacter vinelandii hybrid transformants carrying the unique antigenic determinant suggest that these polysaccharides bind R. trifolii to the clover root hair tips which contain trifoliin.
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Three hundred C3H mice were used to ascertain the validity of treatment of brain cancer with arsenicals and bacterial polysaccharide. It was found that this method of therapy was efficacious. Also, that a prophylactic effect was demonstrated. 2. In 14 patients with advanced intracranial neoplasm it was found: a) that no curative effect could be brought about once the cancer had spread beyond a certain point. This "point of no return" depends on tumor type, location and degree of brain destruction and general state of debility. b) That subjective and even some temporary objective improvement was possible even in advanced cancer. Necrosis of cancer tissue, that could be attributed to the therapy, was found in a number of cases. c) That some cases of brain cancer showed remarkable response to this form of therapy; more so if radiation therapy was given at the time of administration of the arsenical and bacterial polysaccharide. d) That some cases of brain metastasis showed "complete" destruction of the neoplasms in the brain although the patient subsequently died of the primary neoplasm and multiple metastasis. e) That the principle of enhancing the deposition of the curative material in the neoplasm by the use of bacterial polysaccharide is valid. f) That if this method of treatment (i.e. arsenical, bacterial polysaccharide and radiation) is instituted in the "early" cancer cases we may find it to be an efficacious mode of attack. g) That aresnical by mouth and bacterial polysaccharide by I.M. injection may be useful as a prophylactic to the formation of cancer. This may be contemplated for use in families that seem to show a predisposition to cancer formation. A mode of administration would probably be somewhat similary to the maintencance therapy described in the body of this paper. h) That bacterial polysaccharides have been shown to have the ability to destroy cancers.3,9 This method of enhancing the patients antigen-antibody reaction may eventually be used as a means of gaining an efficient vaccine in cancer therapy. i) Wherever possible definitive surgery should be carried out before the arsenical-bacterial polysaccharide-radiation method is instituted. j) In brain cancer, after craniotomy with removal of all or part of the neoplasm where feasible, the patient should be left with a subtemporal decompression. This will allow for the oedema of the brain that occurs with cerebral radiation therapy. k) That the principle of destruction of the cancer by certain special substances is valid. That the increase of affinity between cancer and destructive (curative) material can be brought about by administering a bacterial polysaccharide at the same time and that radiation therapy may enhance the beneficial effects of this method. 1) That the principle of brining the greatest toxicity to the cancer cells and the least toxic effect to the organism has been applied in the use of this method of treatment. m) That in some cases the cancer may not be destroyed by this therapy but may be made to retrogress or be held in check.