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M Rogolsky

Publications and source records attributed to M Rogolsky.

At least 19 recordsLinked to original sources
PubMed1988

Molecular analyses of conjugative, gentamicin-resistance plasmids from staphylococcal clinical isolates.

Gentamicin-resistant Staphylococcus aureus and Staphylococcus epidermidis strains which were isolated from infants with staphylococcal bacteremia were analyzed for the presence of self-transmissible gentamicin-resistance (Gmr) plasmids. Conjugative GMr plasmids of approximately 43.8-63 kilobases (kb) were found in all S. aureus strains. Inter- and intra-species transfer of Gmr plasmids by conjugation was observed from S. aureus to S. aureus and to S. epidermidis recipient strains. However, neither inter- nor intra-species transfer of gentamicin resistance by conjugation was observed with nine out of nine S. epidermidis donor strains which were mated with either S. epidermidis or S. aureus recipient strains. These conjugative Gmr plasmids were unable to comobilize a smaller (15-kb) plasmid present in all but two S. aureus clinical isolates. Many of the conjugative Gmr plasmids also carried genetic determinants for kanamycin, tobramycin, neomycin, and ethidium bromide resistance, and for beta-lactamase synthesis. EcoRI restriction endonuclease digests of the S. aureus Gmr conjugative plasmids revealed three different digestion patterns. Four EcoRI restriction endonuclease digestion fragments of 15, 11.4, 6.3, and 4.6 kb in size were common to all plasmids. These plasmids and conjugative Gmr staphylococcal plasmids from other geographical regions shared restriction digestion fragments of similar molecular weights. DNA hybridization with biotinylated S. aureus plasmid pIZ7814 DNA revealed a high degree of homology among these plasmids. A 50.9-kb plasmid from one of the nonconjugative S. epidermidis clinical isolates showed homology with the probe DNA but lacked a portion of a 6.3-kb fragment which was present in all conjugative plasmids and believed to carry much genetic information for conjugation.

Conjugation, Genetic↗
PubMed1987

Characteristics of coagulase-negative staphylococci from infants with bacteremia.

Twenty-nine infants were identified as having coagulase-negative staphylococcal (C-S) bacteremia. Fourteen infants had pneumonia and 10 had central line-associated bacteremia. Twenty-four of 29 (83%) had invasion of the mucocutaneous barrier at the time the positive blood culture was drawn. Clinical signs and symptoms were nonspecific. Apnea/bradycardia was the most prevalent clinical feature, occurring in 20 (69%) infants. Staphylococcus epidermidis was the most frequent blood culture isolate, occurring in 21 (72%) cases. Slime production by C-S blood culture isolates occurred in 23 (79%) cases. There was no prevalent antibiotic resistance pattern, phage type or plasmid profile among blood culture isolates from infants with bacteremia. Mucocutaneous isolates of C-S from infants with bacteremia were compared with those from infants without invasive disease. Infants with bacteremia had a significantly higher percentage of slime-producing organisms (75% vs. 58%, P = 0.027) and a significantly higher percentage of S. epidermidis species (79% vs. 53%, P = 0.001) than isolates from infants without bacteremia. Our data support the relationship of slime production and the S. epidermidis species of C-S as virulence factors in infants with foreign bodies. Testing C-S for slime production is a relatively simple laboratory procedure which may be an additional aid in the evaluation of their clinical significance.

Bacteriophage Typing↗
PubMed1986

Transfer of the plasmid for exfoliative toxin B synthesis in mixed cultures on nitrocellulose membranes.

Plasmid pRW002 carries genetic determinants for exfoliative toxin B and bacteriocin R1 synthesis. When a donor strain carrying plasmid pRW002 was mixed with a plasmidless recipient strain on a nitrocellulose membrane in accordance with the procedure used for staphylococcal conjugation, pRW002 was passed to the recipient by mixed-culture transduction. Transfer was inhibited by citrate and serotype B phage antisera but not by DNase I. Cell-to-cell contact was not required, and transfer frequencies increased more than 10-fold in the presence of small concentrations of mitomycin C. These results are consistent with pRW002 transfer in mixed cultures by transduction and not by conjugation or transformation. Immunodiffusion and DNA analyses after agarose gel electrophoresis demonstrated that transductants were exfoliative toxin B producers and housed pRW002. Since mixed-culture transfer has been reported to occur on skin, our results suggest that mixed-culture transduction might be a mechanism for the transfer of genetic determinants for pathogenicity in vivo.

Bacterial Toxins↗
PubMed1978

Effect of a staphylococcin on Neisseria gonorrhoeae.

Phage group 2 staphylococcal strain UT0002 contains a large 56S virulence plasmid with genes that code for both exfoliative toxin and a specific staphylococcin termed Bac R(1). Four penicillinase-producing strains and three penicillin-susceptible strains of Neisseria gonorrhoeae were killed by Bac R(1). After 30 min of growth of the penicillin-resistant TR1 strain in 62.5 arbitrary units of Bac R(1) per ml, loss of viability was approximately 90%, and, after 5 h, an approximately 99.99% loss of viability was observed. Lysis did not accompany cell death, and 84% of the Bac R(1) added to the growth medium was adsorbed to the gonococcal cells. The extracellular supernatant fluid from a substrain of staphylococcal strain UT0002 cured of the plasmid for Bac R(1) production had no lethal effect on the gonococcal strains. Bac R(1) was also shown to have bactericidal activity against an L-form of N. meningitidis, indicating that the outer envelope of a neisserial cell is not needed for bacteriocin activity. Ten different normal human sera were unable to neutralize Bac R(1) activity. The bacteriocin lacks adsorption specificity. It binds to but does not kill Escherichia coli cells, indicating that the cell envelope of gram-negative organisms can provide protection against the staphylococcin.

Absorption↗
PubMed1977

Production and properties of a staphylococcin genetically controlled by the staphylococcal plasmid for exfoliative toxin synthesis.

Previous data from this laboratory showed that certain phage group 2 staphylococci contain a large 56S virulence plasmid containing genes that code for both exfoliative toxin (ET) and a specific staphylococcin. Optimal cultural conditions for bacteriocin production were similar to those found for ET production. The bacteriocin is an extracellular product produced in small quantities that can be neither extracted from cell pellets with 1 M NaCl nor induced with mitomycin C. The staphylococcin is active against a wide variety of gram-positive organisms and also against group 2 staphylococcal strains that have been cured of the plasmid carrying the staphylococcin marker. The bacteriocin is not inactivated by oxidation, mechanical agitation, or boiling for 15 min. It is sensitive to the action of trypsin and Pronase but not lysostaphin and is stable within a pH range of 4 to 9. It has an isoelectric point of approximately 7.7. Removal of the ampholytes and glycerol from electrofocused staphylococcin preparations resulted in total loss of bacteriocin activity.

Bacteriocins↗
PubMed1977

Molecular and serological differentiation of staphylococcal exfoliative toxin synthesized under chromosomal and plasmid control.

Evidence for the existence of two molecular species of exfoliative toxin (ET) synthesized by phage group II Staphylococcus aureus under chromosomal and plasmid control is presented. Serological evidence that these molecular species of toxin are distinct from each other is given. The plasmid-controlled toxin was synthesized along with the chromosomally controlled toxin by the group II UT0002 strain, whereas another group II strain, UT0007, synthesized only the plasmid-controlled toxin. The molecular weight of the plasmid-controlled toxin was slightly less than that of the chromosomally controlled type and could be separated from the latter on 12.5% sodium dodecyl sulfate (SDS)-polyacrylamide slab gels. On 7.5% SDS-polyacrylamide cylindrical gels there was no hint of heterogeneity, and both ETs migrated together as a single homogeneous band. The existence of two serotypes of ET among phage group II strains complicates interpretation of previous work in this field and makes necessary the preparation of two different antigens for radioimmunobinding assays. Discovery of these ET serotypes provided an explanation for previously reported low binding by rabbit hyperimmune serum (B. Wiley, L. Glasgow, and M. Rogolsky, Infect. Immun. 13:513-520, 1976) in the radioimmunobinding test. A molecular species of ET differing from each of the other two serotypes was isolated from cultures of a phage group III S. aureus. This ET produced scalding in suckling mice and was lower in molecular weight than the ET produced under plasmid control by group II strains. Preliminary serological studies indicated that the ET in the group III strain is closely related to or possibly identical to the group II toxin produced under plasmid control.

Antigens, Bacterial↗
PubMed1976

Phage group II staphylococcal strains with chromosomal and extrachromosomal genes for exfoliative toxin production.

Staphylococcal phage group 2 strain UT0007 was previously shown to contain a high-molecular-weight plasmid containing genes for exfoliative toxin (ET) and bacteriocin production. Phage group 2 strains UT0002 and UT0003 (Tox+Bac-) underwent a twofold and ninefold loss of ET activity, respectively, after growth at 44 C for 18 h. Strain UT0002 also lost total bacteriocin activity. Both strains contained (i) a 56S plasmid that was lost from those substrains showing reduced ET activity and (ii) a 21S plasmid with a gene for cadmium resistance that could be transduced into two recipient strains. Since the ET plasmid-negative substrains still made ET, it was postulated that this residual toxin was made from chromosomal genes. In characterizing the plasmid species from strains UT0002 and UT0003, the 21S but little or no 56S plasmid deoxyribonucleic acid could be isolated after centrifugation of cleared lysates from these strains on dye-buoyant density gradients. Treatment of cleared lysates from strain UT0002 with ethidium bromide, Pronase, or sodium dodecyl sulfate, but not heat at 60 C, induced conversion of the 56S closed circular ET plasmid to a 38S open circular form as determined after centrifugation on 5 to 20% neutral sucrose gradients.

Cadmium↗
PubMed1976

Staphylococcal scalded-skin syndrome: development of a primary binding assay for human antibody to the exfoliative toxin.

Exfoliative toxin (ET) from a phage group II Staphylococcus aureus strain causing staphylococcal scalded-skin syndrome was purified by electrofocusing. Ampholytes and salts were removed from the final product by column chromatography on G-50 Sephadex. Sodium dodecyl sulfate-polyacrylamide gels of the final product yielded a single band upon gel electrophoresis, even when 60 mug of protein was placed in the gels. Radiolabeling of the purified toxin with 125I yielded a product that still caused exfoliation of suckling mice, indicating that the toxin was still biologically active. A radioimmunobinding assay was developed and used to test rabbit and human sera for antibodies to exfoliative toxin. Although the maximum percentage of binding was not as high as expected (approximately 40%), it was postulated that either iodination had not been sufficiently vigorous or the toxin had sustained immunological damage. The assay was reproducible and more sensitive than the existing neutralization method and readily applicable to the testing of human sera for exfoliative toxin antibodies.

Animals↗
PubMed1975

Chromosomal synthesis of staphylococcal exfoliative toxin.

Tox-+ staphylococcal strains, as opposed to Tox-minus strains, produce epidermal exfoliation within 18 h after direct subcutaneous or intraperitoneal injection into newborn mice. The extracellular product responsible for exfoliation is termed exfoliative toxin (ET). When culture supernatant fluid from the plasmid-cured Tox-minus substrains UT 0100 or UT 0111 or from six naturally occurring phage group 2 Tox-minus strains was concentrated 20-fold and inoculated into newborn mice, ET activity could be detected. The Tox-minus, cured derivatives produced ET at levels which were 32 minus and 64-fold lower than the amounts made by their Tox-+ parent strains. Since these Tox-minus, cured substrains contained no plasmid deoxyribonucleic acid, it was postulated that the product possessing ET activity in strains UT 0100 and UT 0111 was made by chromosomal genes. This product has been isolated and purified from strain UT 0100 and appears as two faint bands after electrophoresis on polyacrylamide gels and corresponds in position to a heavy band of ET isolated from the Tox-+ strain UT 0007.

Animals↗
PubMed1975

Isolation of extrachromosomal deoxyribonucleic acid for exfoliative toxin production from phage group II Staphylococcus aureus.

The ability of phage group II staphylococcal strain UT 0101 to produce exfoliative toxin and bacteriocin could be eliminated at a high frequency after growth at high temperatures or in the presence of ethidium bromide or sodium dodecyl sulfate. Extrachromosomal deoxyribonucleic acid, associated with the genes for exfoliative toxin and bacteriocin production, was isolated from strain UT 0101 but was absent from an ethidium bromide-cured substrain. The molecular weight of the exfoliative toxin plasmid, determined by co-sedimentation with the penicillinase plasmid, PI258, was 3.3 times 10-7. The 56S covalently closed circular form of the exfoliative toxin plasmid converted to a 38S open circular form after storage or exposure to sodium dodecyl sulfate. Plasmid deoxyribonucleic acid associated with penicillin resistance could not be identified in the penicillin-resistance Tox+ strains, UT 0007 and UT 0001.

Bacteriocins↗
PubMed1974

Interaction of staphylococcal exfoliative toxin with concanavalin A.

Treatment of mouse embryo fibroblasts (MEF) grown in vitro with purified staphylococcal exfoliative toxin (ET) increased the concanavalin A (Con A) agglutinability of MEF 3.5-fold over control cells. Possible explanations for this phenomenon were investigated. ET lacked proteolytic activity on denatured casein. Con A, however, was found to interact directly with ET as evidenced by the formation of precipitation in an agar gel diffusion plate and in increased turbidity in solution. This interaction was inhibited by alpha-methyl-d-glucopyranoside. The ability of Con A to precipitate with ET suggests that the toxin contains a carbohydrate component and that the carbohydrate associated with ET is branched rather than linear. An analysis of a purified preparation of ET indicated the presence of 9% carbohydrate and no lipid.

Agglutination Tests↗
PubMed1974

Staphylococcal scalded skin syndrome: potentiation by immunosuppression in mice; toxin-mediated exfoliation in a healthy adult.

Staphylococcal scalded skin syndrome, associated with exfoliative toxin produced by phage group II Staphylococcus aureus, has recently been reported in an adult receiving immunosuppressive therapy. To determine the effect of immunosuppression on the development of the staphylococcal scalded skin syndrome, experimental animals were treated with prednisolone, azathioprine, or a combination of both drugs utilizing the clinical isolate from the adult with scalded skin syndrome. The mean lethal dose and mean exfoliating dose were identical and were 6,000-fold lower in animals receiving both drugs or azathioprine alone. The isolate was not more virulent and did not produce more toxin than other group II phage-type strains. Furthermore, immunosuppressive therapy failed to enhance the susceptibility of experimental animals to a purified preparation of toxin. Finally, purified exfoliative toxin was demonstrated to produce erythema, Nikolsky's sign, bullous formation, and flaking desquamation in a normal human adult. The results demonstrated the enhanced susceptibility of experimental animals receiving immunosuppressive therapy to the development of the staphylococcal scalded skin syndrome. They further showed that human adults are susceptible to the action of exfoliative toxin and suggested that, in the host with compromised defense mechanisms, toxin-producing strains may invade and initiate infection resulting in toxin production and exfoliation.

Adult↗
PubMed1974

Nature of the genetic determinant controlling exfoliative toxin production in Staphylococcus aureus.

Phage group II Staphylococcus aureus has been identified as the etiological agent of the staphylococcal scaleded skin syndrome. The development of an animal model system permitted fulfillment of Koch's postulates and recognition of exfoliative toxin (ET) as being responsible for some of the clinical manifestations of this syndrome. Initial studies directed toward associating a lysogenic phage with the genetic control of ET synthesis failed to support this hypothesis. Growth of two Tox(+) strains at 44 C was more effective than growth in ethidium bromide or sodium dodecyl sulfate in eliminating the ability to produce ET. The early and rapid accumulation of ET-negative (Tox(-)) variants during growth of strain UT 0007 at 44 C, the lack of any selective advantage of the Tox(-) variants over Tox(+) cells during growth at 44 C, and an enhanced elimination frequency at 44 C of 97.9% over the spontaneous frequency of loss strongly suggest that the gene for ET synthesis is extrachromosomal. Additional evidence suggests that this gene is located on a plasmid which is not associated with genes for penicillinase synthesis and cadmium resistance. Two Tox(+) strains harbored lysogenic phage capable of transducing cadmium resistance, but not penicillin resistance, to specific Tox(-) recipients.

Animals↗
PubMed1974

Effect of ethidium bromide on elimination of exfoliative toxin and bacteriocin production in Staphylococcus aureus.

The scalded skin syndrome has been associated with phage group II staphylococci. The clinical manifestations of scalded skin syndrome, Ritter's disease, scarlatiniform erythema, and localized bullous impetigo, are due to the production of an extracellular protein, designated exfoliative toxin. Phage group II staphylococci can also produce an extracellular protein, bacteriocin, which is bacteriocidal for specific gram-positive microorganisms. Strain UT 0007 produces both bacteriocin and exfoliative toxin which appear to be products of extrachromosomal genes. These genes are jointly eliminated from strain UT 0007 after growth in either ethidium bromide or at high temperatures.

Animals↗
PubMed1974

Sensitivity of an early step in the sporulation of Bacillus subtilis to selective inhibition by ethidium bromide.

When a final concentration of 0.4 mug of ethidium bromide (EB) per ml, which is subinhibitory to vegetative growth, is added to sporulating cells of Bacillus subtilis Marburg during either stage 0 or the early part of stage 1, morphogenesis is blocked. If the given concentration of EB is added after the early part of stage 1, sporogenesis is unaffected. The synthesis of the serine protease and antibiotic, which are believed to be associated with sporulation events during the early part of stage 0, are not inhibited by EB. Enhanced binding of [(14)C]benzylpenicillin to sporulating cells during septation (stage 2) is a measure of the presence of terminal enzymes for germ cell wall peptidoglycan synthesis. EB does not interfere with the binding of penicillin to sporulating cells, but penicillin remains more permanently bound to EB-treated postlogarithmic cells than to untreated sporulating cells. The absence of an interval of increased penicillin binding activity during stage 2 by sporulating cells treated with EB indicates that EB blocks sporulation prior to the completion of the germ cell wall.

Anti-Bacterial Agents↗
PubMed1973

Binding of radioactive benzylpenicillin to asporogenous mutants of Bacillus subtilis during postexponential growth.

The specific penicillin binding capacity of a postexponential culture of Staphylococcus aureus remains constant, but that of a sporulating Bacillus subtilis culture fluctuates dramatically. An initial decrease in binding capacity during presporulation events is followed by two distinct intervals of enhanced specific binding capacity during the postlogarithmic growth of a sporulating B. subtilis culture. The first peak of enhanced binding occurs during septation, when enzymes for germ cell wall formation are present; and the second peak coincides with cortical biosynthesis. The specific postlogarithmic binding capacities of a number of Spo(-) mutants of B. subtilis were examined to ascertain if specific asporogenous mutations altered the binding pattern observed with the wild-type organism. Four distinct postexponential binding patterns were recognized: (i) a low, constant binding capacity resembling the binding pattern of S. aureus, (ii) a decrease in binding capacity with no subsequent significant peaks, (iii) a decrease in binding capacity followed by a single peak corresponding to the first peak seen with the wild type, (iv) a pattern similar to the wild type. The fourth pattern was observed in a mutant blocked during stage III of sporogenesis which produced forespores that never became refractile. Mutations blocking either one or both periods of enhanced postlogarithmic binding were interspersed throughout a linkage group of spore genes next to lys-2 on the B. subtilis chomosome.

Bacillus subtilis↗