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Biomedical subjects

M L Higgins

Publications and source records attributed to M L Higgins.

89 records · Page 5Linked to original sources

Morphokinetic reaction of cells of Streptococcus faecalis (ATCC 9790) to specific inhibition of macromolecular synthesis: dependence of mesosome growth on deoxyribonucleic acid synthesis.

The application of quantitative electron microscopy to thin sections of cells of Streptococcus faecalis specifically inhibited for deoxyribonucleic acid (DNA), ribonucleic acid, and protein synthesis shows that septal mesosomes (i) increase in size when protein synthesis is inhibited by at least 80% while DNA synthesis proceeds at no less than 50% of the control rate and (ii) decrease in size when DNA synthesis is inhibited 50% or more during the initial 10 min of treatment. This indicates that fluctuations in mesosome size are dependent on the extent of DNA synthesis. The fluctuations in mesosome areas observed on treatment do not correlate with the kinetics of glycerol incorporation per milliliter of a culture. However, when glycerol incorporation is placed on a per cell basis, a strong correlation is observed between increases in (i) the thickness of the electron-transparent layer of the cytoplasmic membrane and (ii) the amount of glycerol incorporated per cell. It seems that the electron-transparent membrane layer may thicken to accommodate changes in lipid content when protein and lipid synthesis are uncoupled.

Anti-Bacterial Agents↗

Reinitiation of cell wall growth after threonine starvation of Streptococcus faecalis.

Cultures of Streptococcus faecalis ATCC 9790 were starved of threonine for 10 hr and then allowed to reinitiate growth in a fresh complete medium. On regrowth, culture turbidity began to increase within 10 min, but the ability of cells to autolyze did not begin to increase until after 30 min. Ultrastructural studies of regrowth of the initially thick-walled cells showed, at about 30 min, centripetal linear extension of new thin cross wall. This was followed, at about 40 min, by a notching, splitting, and peeling apart of the base of the cross wall. After this, extension of new thin peripheral wall from the nascent cross wall appeared to push old thick wall toward the poles. After the first cell division, asymmetric cells with one initial generation thick-walled pole and one second generation thin-walled pole were seen. After two divisions, thick-walled hemispheres were still seen, suggesting conservation of old wall during this time. A small fraction of the initial cell population exhibited aberrations and difficulties in reinitiating linear wall extension and were useful in the establishment of a model for the reinitiation of linear wall extension.

Bacteriolysis↗

Motile nocardoid Actinomycetales.

The properties of 42 strains of nocardoid (nocardioform) bacteria were compared. The results indicate that the organism previously called Nocardia turbata does not belong to the genus Nocardia nor does it fit into any of the previously described genera.

Actinomycetales↗

Model for cell wall growth of Streptococcus faecalis.

In exponentially growing and dividing cells of Streptococcus faecalis, it is proposed that the leading edge of the annularly closing cross wall is the point of extension for both cross wall and peripheral wall. Peripheral wall extension is thought to be produced by the separation or splitting of the cross wall at its junction with peripheral wall. This results in the pushing of the equatorial wall bands, found on S. faecalis walls, to subsequatorial positions. These bands therefore mark the separation of old wall from new wall. Mesosomal formation was observed usually to precede cross wall initiation.

Cell Wall↗

Early changes in the ultrastructure of Streptococcus faecalis after amino acid starvation.

Thin sections of Streptococcus faecalis (ATCC 9790) starved of one essential amino acid (threonine or valine) initially show rapid increases in (i) cell wall thickness, (ii) the apparent size of the central nucleoid region, and (iii) mesosomal membranes. The most rapid increases in all three variables occurred during the first 1 to 2 hr of starvation. After this initial period, the rates progressively decreased over the 20-hr observation period. During threonine starvation, the mesosomal membrane that accumulated in the first hour was subsequently degraded and reached a level similar to that found in exponential-phase cells after 20 hr. With valine starvation, mesosomal membrane continued to slowly accumulate over the entire 20-hr observation period. The mesosomes of the starved cells retained the same "stalked-bag" morphology of those in exponential-phase cells. These cytological observations agree with previously published biochemical data on membrane lipid and wall content after starvation.

Bacteriological Techniques↗

Site of initiation of cellular autolysis in Streptococcus faecalis as seen by electron microscopy.

Low concentrations of glutaraldehyde (0.1% or higher) blocked cellular and wall autolysis. The site of autolytic activity was studied by allowing cell autolysis to proceed for very short periods (0 to 15 min) before addition of glutaraldehyde. Electron microscopy of ultrathin sections showed that the primary site of autolytic activity was the leading edge of the nascent cross wall. The base of the cross wall seemed more resistant than the tip. Evidence supporting the involvement of autolysin activity in continued wall extension and in cell separation as well as in the initiation of new sites of wall extension was obtained. In cells exposed for 10 min to chloramphenicol, wall dissolution was very much slower but occurred at the same cross wall site.

Autolysis↗

Poorly lytic bacteriophage from Dactylosporangium thailandensis (Actinomycetales).

Dactylosporangiophage A1 has a polygonal head (75 nm) with spherical capsomeres (3 nm) and a noncontractile tail (200 by 10 nm) with cross-striations which is terminated with at least three prongs which are used for attachment. It contains double-stranded deoxyribonucleic acid and produces very little lysis. Intracellular phage multiplication leads to the formation of crystalline aggregates of apparently complete virions. Plaques are formed only on certain substrains of Dactylosporangium thailandensis L1 and are always small (0.5 mm or less). They are clear on some substrains and turbid on others. Formation of plaques occurs only on one medium, Czapek agar with 0.2 to 0.4% yeast extract, 0.2% peptone, or a defined mixture of amino acids. Over 100 strains of bacteria, mainly actinomycetes, were screened in a futile attempt to find an indicator strain which is not a substrain of L1. The Dactylosporangium-phage system studied is considered to be a semiresistant carrier state.

Actinomycetales↗

Flagellated actinomycetes.

Shadowed motile elements from actinomycetes were observed with an electron microscope. Included were three strains of Actinoplanes, two of Ampullariella, two of Dermatophilus, two of Spirillospora, and four of "Nocardia" turbata. In addition, three types of previously undescribed actionmycetes were represented: (i) the C(4) group (four strains) forming substrate mycelium breaking into motile rods; (ii) strain 9-41, forming Microellobosporia-like sporangia with motile spores; and (iii) strain P(2), forming aerial hyphae releasing motile cocci when put in water. All the known chemical cell wall types of actinomycetes except the Nocardia asteroides type and the Actinomyces israeli type were represented in this array of motile actinomycetes. Motile elements were, depending on the genus, cocci, rods (often curved), or pyriform. Flagella were always in tufts (or single), never peritrichous. A relationship seems to exist between the location of the tuft and the cell wall composition. The spores of one strain of Actinoplanes were herniated, thus resembling plasmoptysis forms of bacteria.

Actinomycetales↗

Release of sporangiospores by a strain of Actinoplanes.

Dehiscence of Actinoplanes sp. 7-10 sporangia is triggered by wetting of the spores. This process requires time because of the hydrophobic nature of the sporangial envelope; it can be speeded up and enhanced by a wetting agent. Once wetted, the spores swell, usually ripping the sporangial wall, and escape as motile elements when functional flagella are synthesized. Flagellation and motility are separate phenomena, both of which lose intensity with age. Spores from old sporangia can regain motility when supplied with an exogenous carbon source, but, when provided only with water, phosphate buffer, or amino acids, flagellation takes place without motility. Deflagellation-reflagellation experiments indicated that functional flagella can be reformed only in presence of both amino acids and glucose which must be added within 180 min of deflagellation. Inoperative flagella were formed in the presence of inhibitors of nucleic acid synthesis, such as 6-azauracil, but inhibitors of protein synthesis, such as chloramphenicol, did not interfere with reflagellation. Flagellated spores remained so after germination.

Actinomycetales↗

Analysis of nutritional shift-up of Streptococcus faecium.

Three-dimensional reconstruction methods were applied to electron micrographs of Streptococcus faecium to study the initiation of cell wall growth sites during a nutritional shift-up experiment. Upon lowering the mass doubling time from 76 to 33 min by the addition of excess glutamate, the formation of new cell wall growth sites was studied in relation to other growth parameters (autolytic capacity, cell number, mass, RNA, DNA and peptidoglycan). The findings from these studies, to be described below, support a model in which new sites are introduced when cells grow to a relatively constant, growth-rate-independent size, while the rate at which sites form and grow increases with growth rate. In this model, chromosome synthesis does not regulate the formation of new sites of cell wall growth, but existing sites cannot be completed until rounds of chromosome synthesis are completed.

Cell Wall↗