Structural changes in cells of Clostridium perfringens infected with a short-tailed bacteriophage

1971 ◽  
Vol 17 (3) ◽  
pp. 397-402 ◽  
Author(s):  
David E. Bradley ◽  
Judith F. M. Hoeniger
Keyword(s):  
Electron Microscope ◽  
Clostridium Perfringens ◽  
Structural Changes ◽  
Cell Envelope ◽  
Cell Lysis ◽  
Cell Periphery ◽  
Normal Cells ◽  
Phage Growth ◽  
Cooked Meat ◽  
In Cells

A culture of Clostridium perfringens strain 80 in cooked meat broth contained three types of cell: normal, long, and small round cells. The process of phage maturation and cell lysis was studied in all three forms using the electron microscope. In normal cells, the nucleoplasm first enlarged, then small clear areas, slightly larger than intracellular phages, appeared mainly around the cell periphery. The nucleoplasm dispersed and mature phage particles were formed. Subsequent lysis was caused by the breaking off of portions of the cell envelope leaving holes for the release of the contents. Long cells were also able to support phage growth.

Evaluation of single-electron movies of glutamine synthetase molecules

1983 ◽  
Vol 41 ◽  
pp. 280-281
Author(s):  
W. Kunath ◽  
E. Zeitler ◽  
M. Kessel
Keyword(s):  
Electron Microscope ◽  
Glutamine Synthetase ◽  
Electron Irradiation ◽  
Structural Damage ◽  
Structural Changes ◽  
Single Electron ◽  
Continuous Series ◽  
Digital Recording ◽  
Recording Device ◽  
Low Exposure

The features of digital recording of a continuous series (movie) of singleelectron TV frames are reported. The technique is used to investigate structural changes in negatively stained glutamine synthetase molecules (GS) during electron irradiation and, as an ultimate goal, to look for the molecules' “undamaged” structure, say, after a 1 e/Å2 dose.The TV frame of fig. la shows an image of 5 glutamine synthetase molecules exposed to 1/150 e/Å2. Every single electron is recorded as a unit signal in a 256 ×256 field. The extremely low exposure of a single TV frame as dictated by the single-electron recording device including the electron microscope requires accumulation of 150 TV frames into one frame (fig. lb) thus achieving a reasonable compromise between the conflicting aspects of exposure time per frame of 3 sec. vs. object drift of less than 1 Å, and exposure per frame of 1 e/Å2 vs. rate of structural damage.

The Reaction of Brain Structures with OsO4-Zinc-Iodide Stain

1971 ◽  
Vol 29 ◽  
pp. 272-273
Author(s):  
Werner J. Niklowitz
Keyword(s):  
Electron Microscope ◽  
Structural Changes ◽  
Mossy Fiber ◽  
Toxic Metal ◽  
Staining Technique ◽  
Brain Structures ◽  
Oxidase Inhibitor ◽  
Fiber Layer ◽  
Hippocampal Tissue ◽  
Zinc Iodide

After intoxication of rabbits with certain substances such as convulsant agents (3-acetylpyridine), centrally acting drugs (reserpine), or toxic metal compounds (tetraethyl lead) a significant observation by phase microscope is the loss of contrast of the hippocampal mossy fiber layer. It has been suggested that this alteration, as well as changes seen with the electron microscope in the hippocampal mossy fiber boutons, may be related to a loss of neurotransmitters. The purpose of these experiments was to apply the OsO4-zinc-iodide staining technique to the study of these structural changes since it has been suggested that OsO4-zinc-iodide stain reacts with neurotransmitters (acetylcholine, catecholamines).Domestic New Zealand rabbits (2.5 to 3 kg) were used. Hippocampal tissue was removed from normal and experimental animals treated with 3-acetylpyridine (antimetabolite of nicotinamide), reserpine (anti- hypertensive/tranquilizer), or iproniazid (antidepressant/monamine oxidase inhibitor). After fixation in glutaraldehyde hippocampal tissue was treated with OsO4-zinc-iodide stain and further processed for phase and electron microscope studies.

scholarly journals Signal-mediated localization of Candida albicans pheromone response pathway components

2020 ◽  
Author(s):  
Anna Carolina Borges Pereira Costa ◽  
Raha Parvizi Omran ◽  
Chris Law ◽  
Vanessa Dumeaux ◽  
Malcolm Whiteway
Keyword(s):  
Candida Albicans ◽  
Map Kinase ◽  
Nuclear Localization ◽  
Map Kinases ◽  
Critical Role ◽  
Cell Periphery ◽  
Pheromone Response ◽  
Pheromone Response Pathway ◽  
Localization Signal ◽  
In Cells

Abstract Candida albicans opaque cells release pheromones to stimulate cells of opposite mating type to activate their pheromone response pathway. Although this fungal pathogen shares orthologous proteins involved in the process with Saccharomyces cerevisiae, the pathway in each organism has unique characteristics. We have used GFP-tagged fusion proteins to investigate the localization of the scaffold protein Cst5, as well as the MAP kinases Cek1 and Cek2, during pheromone response in C. albicans. In wild-type cells, pheromone treatment directed Cst5-GFP to surface puncta concentrated at the tips of mating projections. These puncta failed to form in cells defective in either the Gα or β subunits. However, they still formed in response to pheromone in cells missing Ste11, but with the puncta distributed around the cell periphery in the absence of mating projections. These puncta were absent from hst7Δ/Δ cells, but could be detected in the ste11Δ/Δ hst7Δ/Δ double mutant. Cek2-GFP showed a strong nuclear localization late in the response, consistent with a role in adaptation, while Cek1-GFP showed a weaker, but early increase in nuclear localization after pheromone treatment. Activation loop phosphorylation of both Cek1 and Cek2 required the presence of Ste11. In contrast to Cek2-GFP, which showed no localization signal in ste11Δ/Δ cells, Cek1-GFP showed enhanced nuclear localization that was pheromone independent in the ste11Δ/Δ mutant. The results are consistent with CaSte11 facilitating Hst7-mediated MAP kinase phosphorylation and also playing a potentially critical role in both MAP kinase and Cst5 scaffold localization.

scholarly journals Incidence and contamination level of Clostridium perfringens in meat and meat products sold in Sakarya province of Turkey

2021 ◽  
Vol 7 (3) ◽  
pp. 172-178
Author(s):  
Serap Coşansu ◽  
Şeyma Şeniz Ersöz
Keyword(s):  
Clostridium Perfringens ◽  
Meat Product ◽  
Meat Products ◽  
Ground Beef ◽  
Chicken Meat ◽  
Contamination Level ◽  
Biochemical Tests ◽  
Contamination Levels ◽  
Emulsified Meat ◽  
Cooked Meat

Totally 101 meat and meat product samples obtained from local markets and restaurants were analyzed for incidence and contamination level of Clostridium perfringens. The typical colonies grown anaerobically on Tryptose Sulfite Cycloserine Agar supplemented with 4-Methyliumbelliferyl (MUP) were confirmed by biochemical tests. Forty-eight of the samples (47.5%) were contaminated with C. perfringens. The highest incidence of the pathogen was determined in uncooked meatball samples (72.2%) followed by ground beef samples (61.3%). The incidence of C. perfringens in chicken meat, cooked meat döner, cooked chicken döner and emulsified meat product samples were 33.3, 33.3, 28.6 and 16.7%, respectively. Thirteen out of 101 samples (12.9%) yielded typical colonies on TSC-MUP Agar, but could not be confirmed as C. perfringens. Average contamination levels in sample groups ranged from 8.3 to 1.5×102 cfu/g, with the highest ground beef and the lowest chicken meat.

scholarly journals The mechanism of thrombin-induced platelet factor 4 secretion

Blood ◽  
1980 ◽  
Vol 55 (4) ◽  
pp. 661-668 ◽  
Author(s):  
MH Ginsberg ◽  
L Taylor ◽  
RG Painter
Keyword(s):  
Electron Microscope ◽  
Platelet Factor 4 ◽  
Immunofluorescent Staining ◽  
Cell Periphery ◽  
Ultrastructural Examination ◽  
Platelet Factor ◽  
Intact Cells ◽  
Thin Sections ◽  
Intracellular Translocation

We have measured thrombin-induced secretion of platelet factor 4 antigen (PF4) and simultaneously followed its intracellular translocation by immunofluorescence. In permeable resting platelets, speckled intracellular immunofluorescent staining for PF4 was observed. Addition of thrombin to washed platelets at 22 degrees C resulted in secretion of PF4 and formation of large (approximately 0.5 micrometer) immunofluorescent masses. These masses moved to the cell periphery during secretion and were virtually absent at the conclusion of secretion. Ultrastructural examination of thrombin-treated platelets revealed vacuoles corresponding in size, shape, and time of occurrence to the large immunofluorescent masses of PF4. These vacuoles contained PF4 by immunoferritin staining of frozen thin sections; they therefore appear to represent the ultrastructural counterpart of the large PF4 masses. When intact cells were stained for PF4 after thrombin addition, only 5.6% of the large masses stained. Thus, during secretion, PF4 antigen is consolidated into large closed pools that appear as vacuoles in the electron microscope.

scholarly journals Immunocytochemical evidence for a possible role of cross-linked keratinocyte envelopes in stratum corneum cohesion.

1991 ◽  
Vol 39 (11) ◽  
pp. 1531-1538 ◽  
Author(s):  
M Haftek ◽  
G Serre ◽  
V Mils ◽  
J Thivolet
Keyword(s):  
Stratum Corneum ◽  
Cell Envelope ◽  
Ultrastructural Localization ◽  
Antigenic Determinants ◽  
Cell Periphery ◽  
Protein Distribution ◽  
Cell Structures ◽  
Physical Resistance ◽  
Cell Envelopes

Cross-linked cornified envelopes are cell structures specifically synthesized by terminally differentiating keratinocytes. They are composed of proteins deposited at the cell periphery under the plasma membrane, and can be purified from epidermis by physicochemical extractions. The resulting keratinocyte "shells" are highly insoluble structures devoid of cytoplasmic components. The rigidity of the stratum corneum cell envelope seems to be one of the essential factors contributing to the physical resistance of this most superficial epidermal layer. We studied the purified cell envelopes from human plantar horny layer to determine their antigenic composition and protein distribution. The extraction protocol consisted of four 10-min cycles of boiling in 10 mM Tris-HCl buffer containing 2% SDS and 1% beta-mercaptoethanol. The absence of any extractable proteins persisting in the purified pellets was checked with SDS-PAGE of the sample electroeluates. Indirect immunofluorescence as well as pre- and post-embedding immunogold labeling for electron microscopy revealed the persistence of several keratinocyte antigenic determinants on the purified substrates. The antibodies directed against involucrin, keratin 10, desmoplakin I + II, desmoglein (intracellular epitope), intercellular corneodesmosome proteins, and filaggrin (a considerably weaker reactivity) labeled the cell envelopes according to the ultrastructural localization pattern characteristic for a given antigen. We conclude that the cytoskeletal and desmosomal components become "embedded" in the highly cross-linked cornified envelope structures during the process of keratinocyte terminal differentiation. This underlines the central role of cornified envelopes in the physical resistance of superficial epidermal layers and indicates a possible importance of junctional proteins in this function.

Ultrastructural Changes in Euglena After Ultraviolet Irradiation

1973 ◽  
Vol 13 (3) ◽  
pp. 799-809
Author(s):  
A. MICHAELS ◽  
A. GIBOR
Keyword(s):  
Ultraviolet Light ◽  
Ultraviolet Irradiation ◽  
Euglena Gracilis ◽  
Structural Changes ◽  
Ultrastructural Changes ◽  
Electron Microscopic ◽  
Bleaching Process ◽  
In Cells

The structural changes associated with the ultraviolet-induced bleaching of light-grown cells of Euglena gracilis were investigated. Our light- and electron-microscopic observations of the bleaching process indicate that there is a continuity of plastid structure in cells 5 generations after receiving a bleaching dose of ultraviolet light. There seems to be a continuous dilution of the plastid thylakoids and a decrease in plastid size in the bleaching cells. There also seems to be a change in the position of the plastids in relation to the mitochondria in the bleaching cells. The plastids and possibly the mitochondria are the only organelles which are affected by the ultraviolet irradiation. The continuity of plastids in bleaching cells of Euglena is discussed in relation to the proposed effect of the ultraviolet light.

Levels and Enterotoxigenicity of Clostridium perfringens in Pozole, Tamales, and Birria

2005 ◽  
Vol 68 (2) ◽  
pp. 331-335 ◽  
Author(s):  
V. NAVARRO-HIDALGO ◽  
E. CABRERA-DÍAZ ◽  
H. ZEPEDA ◽  
L. MOTA DE LA GARZA ◽  
A. CASTILLO ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Cell Elongation ◽  
Cytotoxic Effect ◽  
Cell Lysis ◽  
Vero Cells ◽  
Plate Count ◽  
Most Probable Number ◽  
Probable Number ◽  
Quantitative Survey ◽  
Plate Counts

A quantitative survey of Clostridium perfringens in typical foods served at local restaurants was conducted for 18 months in Guadalajara, Mexico. A total of 151 samples, including goat's birria (50), pozole (50), and beef tamales (51), were collected from small restaurants in Guadalajara. Samples were tested for C. perfringens by the most probable number (MPN) method and for mesophilic aerobic plate counts (MAPCs) and coliform, yeast, and mold counts by plate count methods. Isolates confirmed as C. perfringens were further sporulated and tested for cytotoxic or cytotonic effect against Vero cells as an indication of enterotoxin production. C. perfringens was detected in 78 (52%) of all samples at concentrations that ranged from 2.3 to 5.4 log MPN/g. Average MAPCs were 1.3 to 2.7 log CFU/g, depending on the type of dish. Coliform counts ranged from less than 1.0 to 1.5 CFU/g, and yeast and mold counts were less than 1.0 log CFU/g in all cases. A total of 118 isolates of C. perfringens were tested for enterotoxic effect on Vero cells; 82 (70%) showed activity against Vero cells. Of them, 31 isolates induced cell lysis, indicating cytotoxic effect; 41 induced cell elongation, indicating cytotonic effect; and 10 produced both cytotoxic and cytotonic effect. Dilution of the bacterial filtrates that were still producing an effect on Vero cells ranged from 1:80 to 1:5,120. These results underscore the importance of determining enterotoxigenicity when testing for C. perfringens in foods.

scholarly journals The endogenous galactofuranosidase GlfH1 hydrolyzes mycobacterial arabinogalactan

2020 ◽  
Vol 295 (15) ◽  
pp. 5110-5123 ◽  
Author(s):  
Lin Shen ◽  
Albertus Viljoen ◽  
Sydney Villaume ◽  
Maju Joe ◽  
Iman Halloum ◽  
...  
Keyword(s):  
Cell Wall ◽  
Structural Changes ◽  
Cell Envelope ◽  
Wall Structure ◽  
Tubercle Bacillus ◽  
Bioinformatics Analyses ◽  
The Past ◽  
Biochemical Assays ◽  
Mycobacterial Protein

Despite impressive progress made over the past 20 years in our understanding of mycolylarabinogalactan-peptidoglycan (mAGP) biogenesis, the mechanisms by which the tubercle bacillus Mycobacterium tuberculosis adapts its cell wall structure and composition to various environmental conditions, especially during infection, remain poorly understood. Being the central portion of the mAGP complex, arabinogalactan (AG) is believed to be the constituent of the mycobacterial cell envelope that undergoes the least structural changes, but no reports exist supporting this assumption. Herein, using recombinantly expressed mycobacterial protein, bioinformatics analyses, and kinetic and biochemical assays, we demonstrate that the AG can be remodeled by a mycobacterial endogenous enzyme. In particular, we found that the mycobacterial GlfH1 (Rv3096) protein exhibits exo-β-d-galactofuranose hydrolase activity and is capable of hydrolyzing the galactan chain of AG by recurrent cleavage of the terminal β-(1,5) and β-(1,6)-Galf linkages. The characterization of this galactosidase represents a first step toward understanding the remodeling of mycobacterial AG.

scholarly journals Inhibitors of complement derived from the erythrocyte membrane in paroxysmal nocturnal hemoglobinuria

Blood ◽  
1980 ◽  
Vol 55 (5) ◽  
pp. 772-776 ◽  
Author(s):  
C Chua ◽  
EM Hoffmann ◽  
JP Adams ◽  
WF Rosse
Keyword(s):  
Erythrocyte Membrane ◽  
Complement Activation ◽  
Inhibitory Activity ◽  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Cell Lysis ◽  
Red Cells ◽  
Cobra Venom ◽  
Normal Cells ◽  
Complement Lysis

Abstract Extracts of the membranes of normal red cells and red cells from all subpopulations of paroxysmal nocturnal (PNH) red cells inhibited antibody-mediated complement activation. These extracts were shown to accelerate decay of the complement complex. C42, and the relative amount of inhibitory activity was similar in normal and PNH membranes. Inhibitors derived from normal red cells markedly decreased lysis of both PNH and normal cells when antibody was present in excess and complement was limiting. These same inhibitors decreased PNH cell lysis to a much lesser degree when complement was activated with cobra venom or acidified serum. The susceptibility of the PNH cell to complement lysis because of an increased fixation of C3 to its membrane is not due to a difference in membrane-associated accelerator of the decay of the C42 complex.

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