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Biomedical subjects

S F Conti

Publications and source records attributed to S F Conti.

At least 55 records · Page 3Linked to original sources

Ultrastructural changes associated with germination and outgrowth of an appendage-bearing clostridial spore.

The sequence of events occurring during the germination and outgrowth of appendage-bearing spores of Clostridium bifermentans was studied by phase-contrast and electron microscopy. The mature spore was characterized ultrastructurally as having the normal spore components as well as long tubular appendages which orginated from the surface of the spore coat. Spores were incompletely enclosed by a distinctly laminated exosporium which possessed hairlike projections on its outermost layer. During germination, structural changes were observed in the core, core wall, cortex, and spore coat layers. Cortical material was extruded from the spore during outgrowth, which usually occurred from the pole opposite the appendages. The subunits comprising the structure of the appendages and the morphology of the mature appendages were observed. No discernible changes could be observed in the spore appendages during germination and outgrowth.

Bacteriological Techniques↗

Facultatively parasitic strain of Bdellovibrio bacteriovorus.

A strain of Bdellovibrio bacteriovorus (designated strain UKi2) was isolated which was capable of growing either saprophytically in host-free medium or endoparasitically in Escherichia coli B/r. It was quantitatively determined that each bdellovibrio could develop in solid medium to produce a colony, and 65% of the cells in a late exponential-phase culture were capable of inducing E. coli B/r spheroplasts. A photomicrographic sequence of single E. coli spheroplasts containing bdellovibrios demonstrated that parasitically derived B. bacteriovorus UKi2 could develop saprophytically after release from the host cells. Strain UKi2 appears to be morphologically quite similar to previously described obligately parasitic bdellovibrios; biochemical data on this strain suggests its close relationship to some of the previously described host-independent strains of Bdellovibrio.

Bacteria↗

Ultrastructure and cell division of a facultatively parasitic strain of Bdellovibrio bacteriovorus.

Some aspects of cell development and division of Bdellovibrio bacteriovorus strain UKi2 were examined by use of electron microscopic techniques. Under saprophytic and parasitic conditions of growth, the comma-shaped cells enlarge, elongate, and form helical filaments. The mechanism of division appears to consist of an asymmetrical constriction of the filamentous cell by the cytoplasmic membrane, accompanied by a breakdown of the outer layers of the cell wall in the division region. During regeneration of the cell wall, the flagellum and flagellar sheath are formed. The development of the flagellum of the daughter cell is initiated prior to separation of the newly formed cells from the filament. Observations of B. bacteriovorus UKi2 grown under saprophytic and parasitic conditions indicate that development and ultrastructure are similar in both modes of growth.

Bacteria↗

Polar lipids of Chromatium strain D grown at different light intensities.

The total amount and the composition of polar lipids of the photosynthetic bacterium Chromatium strain D grown in high (7,500 ft/c) and low (100 ft/c) incident white light were measured. Cells grown with low incident light as compared with cells grown in high incident light contained approximately 3.5 times more chlorophyll and contained significantly more internal membranes (chromatophores) as determined by electron microscopy. Nevertheless, there was approximately 40% more phospholipid in cells grown at 7,500 ft/c than in cells grown with 100 ft/c; the phospholipid composition of the cells was similar under both conditions of growth. These observations lead us to suggest that the organization and perhaps localization of the phospholipids in Chromatium may be dependent on the light intensity supplied during growth.

Carbon Isotopes↗

Isolation of a bacteriophage for Bdellovibrio bacteriovorus.

A phage infective for Bdellovibrio bacteriovorus was isolated. Electron microscopy revealed that it is tail-less, has a hexagonal appearance and two distinct capsomere layers, and is 60 to 70 nm in size. The nucleic acid appears to be single-stranded deoxyribonucleic acid. This is the first report of the isolation of a phage infective for B. bacteriovorus.

Acridines↗

Microgermination of Bacillus cereus spores.

The biphasic nature of germination curves of individual Bacillus cereus T spores was further characterized by assessing the effects of temperature, concentration of germinants, and some inorganic cations on microgermination. Temperature was shown to affect both phases of microgermination as well as the microlag period, whereas the concentration of l-alanine and supplementation with adenosine exerted a significant effect only on the microlag period. The germination curves of individual spores induced by inosine were also biphasic and resembled those of spores induced by l-alanine. High concentrations (0.1 m or higher) of calcium and other inorganic cations prolonged both phases of microgermination, particularly the second phase, and had a less pronounced effect on the microlag period. The second phase of microgermination was completely inhibited when spores were germinated either in the presence of 0.3 m CaCl(2) or at a temperature of 43 C; this inhibition was reversible. Observations on the germination of spore suspensions (kinetics of the release of dipicolinic acid and mucopeptides, loss of heat resistance, increase in stainability, decrease in turbidity and refractility) were interpreted on the basis of the biphasic nature of microgermination. Dye uptake by individual spores during germination appeared also to be a biphasic process.

Alanine↗

Ultrastructure of blue-green algae.

Two freshwater blue-green algae, Tolypothrix tenuis and Fremyella diplosiphon, and an oscillatorialike marine alga, were found to possess structures on the photosynthetic lamellae which appear to correspond to the phycobilisomes of red algae. These homologous structures are important because they contain the phycobilins which are accessory pigments involved in photosynthesis. As in the red algae, the phycobilisomes were attached on the outer side of each lamellae, i.e., the side facing away from its own membrane pair. Although our study on Anacystis nidulans has not thus far revealed the presence of phycobilisomes, some observations were made on the structure of the polyhedral bodies. After negative staining, the polyhedral bodies were seen to be composed of regularly spaced subunits arranged in a crystalline array. Elongated segmented rods, which differed from the polyhedral bodies, were found in the nuclear region of apparently healthy Tolypothrix cells.

Eukaryota↗

Separation and identification of the polar lipids of Chromatium strain D.

The polar lipids of the autotrophically grown, obligately anaerobic, photosynthetic bacterium Chromatium strain D were separated by paper chromatography. Four major phospholipids were identified: lysophosphatidylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin. In addition, three glycolipids were observed and characterized, namely, monoglucosyldiglyceride, which is found in other biological systems, and (mannosyl, glucosyl)-diglyceride and (dimannosyl, glucosyl)-diglyceride, which heretofore have not been observed in nature.

Autoradiography↗

Germination of single bacterial spores.

Changes in refractility and optical density occurring in individual spores of Bacillus cereus T and B. megaterium QM B1551 during germination were investigated by use of a Zeiss microscope photometer. The curves revealed that the germination process in single spores had two distinct phases; an initial rapid phase was followed by a second slower phase. Under the experimental condition employed, the first phase of germination of B. cereus spores lasted for approximately 75 +/- 15 sec, whereas the second phase lasted for 3 to 4.5 min. In B. megaterium spores, the first phase was observed to last for approximately 2 min and the second phase for more than 7 min. The duration of the second phase was dependent on conditions employed for germination. The kinetics of the first phase were strikingly similar under all conditions of physiological germination. Time-lapse phase-contrast microscopy of germinating spores also revealed the biphasic nature of germination. It was postulated that the first phase represents changes induced by an initial partial hydration of the spore and release into the medium of dipicolinic acid, whereas the second phase reflects degradation of the cortex and hydration of the core.

Bacillus cereus↗

Electron microscopic observations on the penetration of Bdellovibrio bacteriovorus into gram-negative bacterial hosts.

The progressive stages in Bdellovibrio bacteriovorus penetration into two strains of Escherichia coli were examined by use of electron microscopic techniques. The initial change observed in the ultrastructure of the host following parasitic attack was the swelling of the cell envelope at the site of attachment. The Bdellovibrio then appeared to pierce the center of this swelling, forming a pore in the outer wall layers of the host. The edges of this entry pore constricted the Bdellovibrio throughout its penetration into the host cell. Although partial disruption of the cytoplasmic membrane was always apparent, the parasite did not appear to actively penetrate through this barrier. An attempt is made to correlate the fine structural changes involved in penetration with the physiological data that have accumulated to date.

Bacterial Physiological Phenomena↗

Effects of oxygen and heme on the development of a microbial respiratory system.

The effect of adding hemin to anaerobically grown cells of a strain of Staphylococcus epidermidis, which was heme-deficient due to anaerobic growth, has been examined. Cells grown anaerobically in media containing hemin exhibited a marked increase in several oxidative activities as compared with cells grown anaerobically without hemin. The respiratory activity of whole cells and a cyamide-sensitive reduced nicotinamide adenine dinucleotide oxidase activity of cell-free extracts were increased fourfold. The content of enzymatically reducible pigments which exhibit difference spectra similar to cytochromes b(1) and o was also markedly increased. These pigments are mostly sedimented at 100,000 x g (1 hr). Hemin also caused a marked increase in respiratory activity when added directly to the anaerobic culture after the period of growth, but did not cause a similar increase in respiration when added to washed, resting-cell suspensions. Under the latter conditions, heme pigments were formed which exhibited difference spectra similar to, but not identical with, the spectra of pigments found in anaerobic cells grown in the presence of hemin. When resting suspensions of cells grown anaerobically without hemin were exposed to air, a rapid fourfold increase in respiratory activity and a limited increase in cytochrome-like pigments occurred. The presence of the heme precursor Delta-aminolevulinic acid during this aeration resulted in a rapid and marked increase in heme pigments, but only a slight stimulation of respiratory activity. The possible implications of these results for the roles which heme and oxygen play in the development of the respiratory system of this organism are discussed.

Cytochromes↗

Granules associated with the chloroplast lamellae of Porphyridium cruentum.

Small granules with a diameter of approximately 350 A are attached to the chloroplast lamellae of the red alga Porphyridium cruentum. To some extent, their size depends on the culture conditions and the age of the cell. It was possible to preserve the granules only with aldehyde prefixation. It can be seen that fixed or negatively stained granules are comprised of smaller subunits. The granules are arranged regularly on the lamellae in repeating rows with a center-to-center granule distance of 400 to 500 A. Attempts at characterization of these chloroplast granules revealed that they are resistant to hydrolysis by ribonuclease and appear to be structurally unaffected by methanol-acetone extraction. Because of their close association with the chloroplast lamellae, they are considered as possible sites of phycobilin concentration. This possibility is supported by two observations: when the phycobilins are removed, the granules disappear; and, when the chlorophyll and stainable membrane portions are selectively removed, the phycobilins and granules are still present. It was found that all other marine red algae examined had granules which were associated with the chloroplast lamellae.

Chloroplasts↗

Respiratory system of Rhodotorula glutinis. I. Inhibitor tolerance and cytochrome components.

Oxygen uptake by the carotenoid-containing yeast, Rhodotorula glutinis was not affected by concentrations of cyanide and antimycin A which completely inhibit the respiration of Saccharomyces cerevisiae. The tolerance of R. glutinis to these inhibitors was somewhat dependent on the age of the cultures. Reduced minus aerated difference spectra of cells revealed spectral changes presumably due to cytochromes and carotenoids. The kinetics of these spectral changes induced by oxygen were followed. Carotenoid deficient cells were prepared by growth in the presence of diphenylamine. Difference spectra of these cells revealed the presence of flavoprotein, and a, b, and c type cytochromes. Growth of R. glutinis was completely inhibited by concentrations of cyanide which did not affect respiration. Oxidation of reduced nicotinamide adenine dinucleotide by sub-cellular fractions was sensitive to cyanide and antimycin A. Although respiration of intact cells is tolerant to these inhibitors, studies with cell-free extracts suggest the presence of a cyanide and antimycin A-sensitive, cytochrome-linked, respiratory chain.

Cytochromes↗

The respiratory system of Rhodotorula glutinis. II. Mechanism of inhibitor tolerant respiration.

The mechanism of inhibitor-tolerant respiration in Rhodotorula glutinis was studied. This inhibitor-tolerant respiratory pathway was not due to the presence of an excess of cytochrome c oxidase, nor to the operation of an inhibitor-resistant cytochrome c oxidase. Carotenoids do not appear to be involved in this respiratory chain pathway; data are also presented which show that the inhibitors penetrate into the cell. Although the initial rate of oxygen uptake by intact cells was not inhibited in the presence of cyanide or antimycin A, in the presence of these inhibitors the rate of oxygen uptake decreased significantly when the oxygen concentration fell below 100 mum. This change in rate of oxygen uptake as a function of pO(2), suggests that a respiratory chain with a low affinity for oxygen operates in the presence of inhibitors. The characteristics of this alternate pathway are described.

Cytochromes↗

Photosynthetic Apparatus in the Green Bacterium Chloropseudomonas ethylicum.

Holt, Stanley C. (Dartmouth Medical School, Hanover, N.H.), S. F. Conti, and R. C. Fuller. Photosynthetic apparatus in the green bacterium Chloro-pseudomonas ethylicum. J. Bacteriol. 91:311-323. 1966.-When cells of Chloro-pseudomonas ethylicum were broken by ballistic disruption and examined by electron microscopy, vesicles 1,300 to 1,500 A long and 300 to 500 A wide were found to rim the periphery of the cell. Examination of these vesicles obtained by disruption with a French pressure cell and purified by density gradient centrifugation revealed inter-connections between the vesicles. During sonic and Mickle disruption of the cells, chlorophyll was released at a lower rate than soluble cytoplasmic components, but faster than the membrane-bound enzyme succinic dehydrogenase. Unlike the situation that exists in the purple photosynthetic bacteria, it appears that the chlorophyll in the green bacteria is contained as part of a structure which may be differentiated both structurally and functionally from the bacterial cytoplasmic membrane.

Journal Article↗