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S C Holt

Publications and source records attributed to S C Holt.

At least 145 records · Page 8Linked to original sources
PubMed1980

Characterization of the lipopolysaccharide from Actinobacillus actinomycetemcomitans Y4 and N27.

The lipopolysaccharide (LPS) from Actinobacillus actinomycetemcomitans strains Y4 and N27 was isolated by the phenol-water procedure. Morphologically, the molecule consisted of ribbon and branched filaments which comprised 3% of the cellular dry weight. Chemical analysis of the isolated and purified LPSs of both strains showed them to consist of carbohydrate, lipid, 2-keto-3-deoxyoctonate, heptose, hexosamine, and phosphate. The major fatty acids of the lipid A moiety were saturated C14 and beta-OH C14 compounds. Rhamnose, fucose, galactose, glucose, heptose, glucosamine, and galactosamine comprised the monosaccharide portion of the LPS. Biological activity studies revealed both LPS molecules to be active in the Schwartzman reaction and in in vitro 45Ca bone resorption, as well as in macrophage activation and lethality and in platelet aggregation.

Actinobacillus↗
PubMed1980

Transmission-scanning electron microscopic observations of selected Eikenella corrodens strains.

The morphology of Eikenella corrodens 333/54-55 (ATCC 23834) and two human periodontal lesion isolates, strains 470 and 373, was examined by transmission and scanning electron microscopy. All strains exhibited a cell envelope characteristic of gram-negative bacteria. Staining with ruthenium red and alcian blue revealed a loosely organized fibrous slime layer associated with the outer surface of the outer membrane. Slime "stabilization" was achieved by incubation of cells with antisera prepared against whole cells of the Eikenella strains. The stabilized slime appeared as a thick, electron-opaque layer juxtaposed to the outer membrane. Negative staining and heavy metal shadow-casting revealed an interwoven network of fibrils approximately 4 nm in diameter. These fibrils appeared to represent subunits of a larger fibril. Scanning electron microscopy after antibody slime stabilization confirmed the presence and location of the slime layer.

Bacteroides↗
PubMed1980

Cellular fatty acid composition of Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus.

Strains of Actinobacillus actinomycetemcomitans isolated from deep pockets of patients with juvenile periodontitis were analyzed for their content of cellular fatty acids. Oral Haemophilus strains, morphologically and biochemically similar to Haemophilus aphrophilus, were also examined for their content of cellular fatty acids. The extractable lipids of the actinobacilli represented approximately 10% of the cell dry weight, with the bound lipids representing 2 to 5%. The major fatty acids consisted of myristic (C14:0) and palmitic (C16:0) acids and a C16:1 acid, possibly palmitoleic acid, accounting for 21, 35, and 31% of the total extractable fatty acids, respectively. Haemophilus strains had a similar cellular fatty acid content.

Actinobacillus↗
PubMed1979

Capnocytophaga: new genus of gram-negative gliding bacteria. I. General characteristics, taxonomic considerations and significance.

The characteristics of gliding bacteria isolated from both healthy and diseased sites in the oral cavity are, summarized and the taxonomic position of the bacteria discussed. Uniform attirubtes of the fusiform isolates include gliding motility, strictly fermentative metabolism dependent on the presence of CO2 (or HCO3-), under either anaerobic or aerobic conditions, presence of benzidine-reactive components, and the production of acetic and succinic acids as the major or sole, acidic, metabolic and products. Given the guanine and cytosine content of DNA, their gliding motility, and the ability of many strains to attack polysaccharide a relationship to the cytophagus is suggested. This relationship, along with the CO2-dependent growth is recognized by the generic name Capnocytophaga given them. Many of the isolates are grouped into three species C. ochracea, C. Sputigena, and C. gingivalis, separated on the basis of morphological and physiological traits.

Adult↗
PubMed1979

Capnocytophaga: new genus of gram-negative gliding bacteria. II. Morphology and ultrastructure.

Gram-negative, anaerobic gliding bacteria were isolated from normal supragingival plaque and from periodontal lesions. Isolates could be divided into two size classes: small 2.4-4.2 micrograms x 0.38-0.5 microgram and large 4.8-5.8 micrograms x 0.42-0.6 microgram cells. The outer membrane was either loose-fitting and wavy, or taut, and of variable thickness. An electron-dense fuzz was discernible on several of the isolates. The periplasmic region was of variable electron-density. The genus Capnocytophaga has been proposed for these organisms based on morphological and cultural characteristics.

Adult↗
PubMed1979

Capnocytophaga: new genus of gram-negative gliding bacteria. III. Physiological characterization.

Sixty-eight strains of capnophilic fusiform Gram-negative rods from the human oral cavity were subjected to extensive physiologic characterization, tested for susceptibility to various antibiotics, and the mol-percent guanine plus cytosine of each isolate determined. The characteristics of the isolates were compared with 10 fresh and 2 stock isolates of Fusobacterium nucleatum. The isolates clearly differed from the Fusobacterium species on the basis of mol-percent guanine plus cytosine, end products, growth in a capnophilic environment and fermentation of carbohydrates. All of the gliding isolates required CO2 and formed acetate and succinate, but not H2S, indole or acetylmethylcarbinol. All fermented glucose, sucrose, maltose and mannose. The organisms may be differentiated on the basis of fermentation of additional carbohydrates, hydrolysis of polymers and reduction nitrate. Three species are proposed: Capnocytophaga ochracea, Capnocytophaga sputigena and Capnocytophaga gingivalis. Ten isolates did not fit into the proposed species.

Adult↗
PubMed1979

Ultrastructure of Bacteroides species: Bacteroides asaccharolyticus, Bacteroides fragilis, Bacteroides melaninogenicus subspecies melaninogenicus, and B. melaninogenicus subspecies intermedius.

Representative strains of two subspecies of Bacteroides melaninogenicus (subspecies melaninogenicus and subspecies intermedius) and Bacteroides asaccharolyticus as well as B. asaccharolyticus strain 536B isolated from a human perirectal abscess and Bacteroides fragilis ATCC 25285 were examined by glutaraldehyde-osmium fixation, ruthenium red fixation and staining, and thorium hydroxyde staining as well as by the physical preparative techniques of critical point drying--transmission electron microscopy (CPD--TEM) and scanning electron microscopy (SEM). All strains, with the exception of B. fragilis 25285, possessed an electron-dense material external to their outer membranes. Ruthenium red staining further revealed a layer, external to the surface of the outer membrane, that was distinct for each species examined. Thorium hydroxide, as well as CPD--TEM and SEM, showed the cells to be interconnected by thin fibers that not only connected adjacent cells but also traversed several microns to connect cell aggregates.

Bacteroides↗
PubMed1979

In vitro studies of dental plaque formation: adsorption of oral streptococci to hydroxyaptite.

A mixture of saliva-coated hydroxyapatite beads and radioactively labeled bacteria has been employed as an in vitro model for the initial phase of dental plaque formation. Adsorption in this model can be expressed by the Langmuir adsorption isotherm, and the adherence of oral streptococci can be expressed as the product of the affinity constant (Ka) and the number of binding sites (N), KaN. With this approach, Streptococcus sanguis serotype 1 strains adhered better (KaN = [187 +/- 72] X 10(-2)) than serotype 2 strains (KaN = [97 +/- 84] X 10(-2)); a t test showed this difference to be statistically significant to the 99.99% confidence level. Strains of S. mitis, S. mutans, and S. salivarius did not appear to adhere as well. To analyze the bacterial receptors involved in adherence, competition studies in which increasing quantities of unlabeled bacteria were added to a fixed quantity (4 X 10(9) cells per ml) of 3H-labeled serotype 1, reference strain S. sanguis G9B, were performed. These studies indicated that the type 1 strains competed for the same, or closely related, binding sites. Competition studies using serotype 2 S. sanguis strains resulted in an increased binding of reference strain G9B to hydroxyapatite. Scanning electron microscopy indicated this effect was due to the formation of localized aggregations of bacteria, presumably representing the two bacterial types. The results of competition studies with S. mitis were variable, and several strains of other oral bacteria showed little or no competition.

Binding Sites↗
PubMed1979

Fatty acid composition of gliding bacteria: oral isolates of Capnocytophaga compared with Sporocytophaga.

The extractable and bound lipids and cellular fatty acids of the gram-negative gliding bacteria, Capnocytophaga sputigena, C. gingivalis, and C. ochracea were compared to the non-host-related gliding bacterium Sporocytophaga myxococcoides. The extractable lipids represented between 17 and 28% of the cell dry weight, whereas only 2 to 4% of the lipids were in the bound fraction. The methyl esters of the cellular fatty acids were mainly aC15:0, which accounted for 69 to 73% of the total extractable fatty acids; S. myxococcoides had a similar distribution of branched-chain fatty acids; however, aC17:0 was the predominant fatty acid in this free-living gliding organism.

Cytophagaceae↗
PubMed1979

Phospholipid composition of gliding bacteria: oral isolates of Capnocytophaga compared with Sporocytophaga.

The distribution of acetone-soluble (neutral glycolipid) and acetone-insoluble (phospholipid isoprenoids) lipids in oral isolates of gram-negative gliding bacteria of the genus Capnocytophaga was compared with those in a non-host-related gliding bacterium, Sporocytophaga myxococcoides. The acetone-soluble material accounted for 34 to 55% of the extracted lipids; the remainder was acetone-insoluble material. The major phospholipid was phosphatidylethanolamine (67%), with lesser amounts of lysophosphatidylethanolamine and several unidentified phosphate-containing compounds. Capnocytophaga also contained significant amounts of an ornithine-amino lipid.

Cytophagaceae↗
PubMed1979

Ultrastructure of gliding bacteria: scanning electron microscopy of Capnocytophaga sputigena, Capnocytophaga gingivalis, and Capnocytophaga ochracea.

When examined by both light and scanning electron microscopy, Capnocytophaga gingivalis, C. sputigena, and C. ochracea displayed three distinct growth zones: the original streak, an intermediate zone, and the advancing edge, or halo zone. On Trypticase (BBL Microbiology Systems)-soy-blood agar, the cells translocated by gliding. C. gingivalis and C. sputigena formed large, irregular isolated colonies, while C. ochracea formed a more confluent cell mass. The cells within the streak zone and in most of the intermediate zone were heaped into mounds, with the individual cells displaying a definite flow pattern, the latter characteristic of C. sputigena and C. gingivalis. The halo zone consisted of tracks of cells which appeared to have translocated back upon themselves, or were restricted in their outward movements by adjacent cells. Also present within the halo zone were small aggregates of cells, referred to as pioneer colonies. The cell surfaces of C. gingivalis and C. ochracea were smooth and free of any apparent extracellular material, whereas C. sputigena was covered with a thick amorphous material, as well as long, thick, cell surface-associated fibrils.

Bacteroidetes↗
PubMed1978

Lectin analysis of Trypanosoma congolense bloodstream trypomastigote and culture procyclic surface saccharides by agglutination and electron microscopic technics.

Living, intact bloodstream trypomastigotes and culture procyclic forms of Trypanosoma congolense were tested for aggulination with the lectins concanavalin A (Con A), phytohemagglutinin P (PP), wheat germ agglutinin (WGA), soybean agglutinin (SBA), and fucose binding protein (FBP). Similar experiments were conducted with living bloodstream and culture forms treated with trypsin or dextranase. Parasites were incubated for 30 min at 25 C in various concentrations of each lectin, then examined for agglutination by dark-field microscopy. Control preparations consisted of parasites incubated alone or with 0.5 M of the specific competing sugar, with or without the corresponding lectin. Electron-microscopic localization of lectin binding sites on the surface of intact and dextranase-treated bloodstream and intact culture forms was accomplished with Con A, reacted with horseradish peroxidase (HRP) and then diaminobenzidine (DAB). In addition, FBP and SBA were coupled to HRP, then utilized for the localization of binding saccharides on the surface of bloodstream forms by the DAB technic. Similar studies were conducted with culture procyclics incubated with WGA-, SBA-, PP- or FBP-HRP conjugates and then reacted with DAB. Controls were utilized to confirm the sugar specificity of all positive reactions. Intact living bloodstream forms were agglutinated in a concentration-dependent manner with all the lectins tested. Agglutination levels were scored as Con A greater than FBP greater than WGA = PP = SBA. Sugars resembling alpha-D-mannose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and alpha-L-fucose are evidently present on the surface of the parasites. No agglutination was noted in any control preparations. Identical lectin-induced agglutinations were obtained with trypsin- or dextranase-treated bloodstream forms. Trypsin disrupted but did not entirely remove the surface coat of bloodstream forms, while dextranase did not alter the ultrastructure of the parasites. Con A-, SBA- and FBP-binding saccharides were distributed uniformly on the surface coat of intact bloodstream forms; a similar distribution of Con A receptors was noted also on the surface of dextranase-treated cells. No lectin-binding saccharides were visualized by electron microscopy on any control preparations. Intact, trypsin- or dextranase-treated, procyclics were agglutinated in a concentration-dependent fashion by Con A and WGA, but not by the other lectins tested. Control preparations did not agglutinate and the enzymes did not affect the ultrastructure of the parasites. Con A- and WGA-specifically binding saccharides were uniformly distributed on intact procyclics and control preparations were lectin-negative. Thus, T. congolense procyclics retained surface saccharides resembling alpha-D-mannose and N-acetyl-D-glucosamine but lost sugars resembling N-acetyl-D-galactosamine (or D-galactose) and alpha-L-fucose...

Agglutination Tests↗
PubMed1978

Characterization of bacteriophage CP1, an organic solvent sensitive phage associated with Pseudomonas cepacia.

Pseudomonas cepacia strain 249 has been found to harbor an organic solvent sensitive phage, CP1, which is active on other P. cepacia strains. The efficiency of plating of CP1 was dependent upon the strain on which it was propagated and the strain used as indicator, implying the operation of host restriction and modification systems in certain of the strains. Strain 383 which was used routinely for propagation of CP1 appears to lack such systems. To obtain high-titer lysates it was important to add EDTA to the infected cultures at the onset of lysis to block attachment of phage particles to cell debris. CP1 possesses a distinct head (55 nm in diameter) and a broad contractile tail (15 x 145 nm). Fluorescent staining of phage preparations with acridine orange indicated that CP1 contains double-stranded DNA. CP1 particles contained about 5 X 10(-17) g each of protein and DNA for a total particle weight of 10(-16) g. The apparent molecular weight of CP1 DNA estimated from its sedimentation behavior and the particle content of DNA was about 3 X 10(7). Thermal-denaturation studies indicated that the G + C content of CP1 DNA (65%) was lower than that of DNA of its P. cepacia host (71% G + C). The mechanism of inactivation of CP1 by chloroform appears to be related to tail contraction caused by this agent. An atypical reverse contraction of the tail sheath was noted in about 45% of the inactivated particles. No phospholipid was detected in purified preparations of CP1 (less than 4 X 10(19) g/PFU). The results suggest that inactivation of CP1 by organic solvents involves alteration of a component (presumably a protein) of the phage tail.

Bacteriophages↗
PubMed1977

Effect of carbon dioxide on pigment and membrane content in Synechococcus lividus.

The effect of carbon dioxide on pigment and membrane content in Synechococcus lividus was studied by depriving cells of CO2 and examining cell populations biochemically and by electron microscopy. After 120 h of CO2 deprivation, S. lividus lost all detectable chlorophyll a and C-phycocyanin. Such bleached cultures were "mustard yellow", the result of approximately 1.8 times more carotenoid per cell than green control cultures. Although cells from beached cultures appeared morphologically identical to control green cells when examined by light microscopy, electron microscopic examination revealed them to be devoid of detectable thylakoid membrane. Thylakoid membrane could not be recovered by physical isolation or revealed by freeze etching of bleached S. lividus. In addition, inclusion bodies characteristically found in S. lividus were also absent. Reintroduction of CO2 into bleached cultures resulted in a rapid resynthesis of both chlorophyll a and C-phycocyanin. Electron microscopic examination of these regreening cultures revealed that thylakoid membrane was also rapidly resynthesized. Growth of regreened cultures did not occur until there was the synthesis of a full complement of chlorophyll a, C-phycocyanin, and thylakoid membrane. A time course study of the cytological events occurring during bleaching and regreening is presented.

Carbon Dioxide↗
PubMed1976

Structure and composition of intracytoplasmic membranes of Ectothiorhodospira mobilis.

The lamellar membrane stacks of Ectothiorhodospira mobilis were isolated and purified by a combination of lysozyme and osmotic shock treatment, followed by differential and density gradient centrifugation. Preparations of lamellar membranes were enriched at least 2.4-fold in the ratio of bacteriochlorophyll a to protein. Thin-sectioning, negative staining, platinum-carbon shadowing and freeze-etching were used to study the architecture of the membrane units. Both platinum-carbon shadowing and freeze-etching showed the outer-surfaces of the isolated lamellar membrane stacks to be relatively smooth. Particles averaging 7 nm in diameter were seen on several faces following freeze-etching.

Amino Acids↗
PubMed1976

Comparative ultrastructure of selected oral streptococci: thin-sectioning and freeze-etching studies.

The ultrastructure of Streptococcus mutans, serotypes a-e, S. sanguis, S. mitis, and S. salivarius HHT, were examined by the techniques of thin-sectioning and freeze-etching. The cell walls varied in width between 15 and 46 nm and were covered with an electron-dense fibrillar or fuzz layer. The cytoplasmic membrane was in close association with numerous mesosomes which were, in turn, either closely associated or in contact with the bacterial chromosome. In freeze-etch replicas, the outermost layer of the cell wall was fibrous, and the cytoplasmic membrane was covered with particles composed of several subunits. Both particle-clusters and particle-free areas occurred on the surfaces of the cytoplasmic membrane, as well as a crystalline array in the ground plasm of the cell.

Cell Membrane↗
PubMed1975

Ultrastructural studies of sporulation in Bacillus sphaericus.

Spore septum formation in Bacillus sphaericus 9602 occurs 2 h after the end of exponential growth at one end of the vegetative cell, which retains a uniform diameter. The apparently rigid spore septum contains an inner cell wall layer which disappears when the sporulation septum "bulges" into the mother cell cytoplasm. This process occurs simultaneously with terminal swelling at the end of the cell containing the spore septum. It is suggested that the inner cell wall layer is peptidoglycan and that its dissolution and the terminal swelling are consequences of a localized autolysis. Engulfment of the forespore by membrane proliferation results in the production of a forespore surrounded by two flexible, closely apposed membranes. These membranes appear to become more rigid as a peptidoglycan-like layer appears between them, concomitant with the condensation of the forespore nucleoid into a crescent-shaped structure. After nuclear condensation, visible development of distinct cortex, primordial cell wall, and spore coat layers begin, and the forespore cytoplasm assumes an appearance similar to that of a refractile spore. The spore coats consist of an amorphous inner layer, a lamellar midlayer, and a structured outer layer. As cortex synthesis and spore coat assembly continue, exosporium development commences close to that portion of the mother cell plasma membrane which surrounds the forespore. The exosporium is lamellar and in tangential section is seen to have a hexagonal arrangement of subunits. The timing of these morphological events has the expected correlation with the appearance of unique enzyme activites required for cortex synthesis.

Bacillus↗