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Biomedical subjects

S Mitsuhashi

Publications and source records attributed to S Mitsuhashi.

At least 289 records · Page 16Linked to original sources

Natural hemolytic activity of snake serum. II. Heterogeneity of putative natural antibody and levels of hemolytic activity.

The erythrocytes of various vertebrates, such as mice, rabbits, sheep, chickens, bullfrogs, and toads are lysed by normal snake sera. However, snake erythrocytes were not lysed by serum from different snake species. Putative natural antibody seems with different specificities to comprise heterogeneous antibodies. Thus, absorption of snake serum with mouse erythrocytes, for example, abrogated hemolytic activity for mice but not for rabbit or sheep erythrocytes. We observed no significant intraspecies individual differences in serum hemolytic titer, but interspecies differences were obvious. Immunization of snakes with sheep erythrocytes caused no further elevation of hemolytic activity, though high titer antibody was produced in response to certain bacterial antigens. Even the sera of newly-hatched snakes showed hemolytic activity at modestly high levels. No seasonal change in hemolytic activity was observed.

Absorption↗

In vivo formation of transmissible resistance factor by recombination between nontransmissible resistance factor and Col B factor.

Germ-free swine were artificially contaminated with tetracycline (TC) sensitive strains of Escherichia coli and Klebsiella pneumoniae. One of these strains, E. coli 3306, was infected with a plasmid carrying kanamycin (KM) resistance, i.e., T-kan factor. Another strain, E. coli P-5, carried a conjugally transferable Col B factor. Among the nine strains used, only E. coli P-38 became TC-resistant after TC administration. Three types of TC-resistance E. coli P-38 strains were found; (a) one strain carried nontransferable TC resistance and could not produce colicin, (b) one strain carried TC resistance with a high transmission frequency which could not produce colicin, and (c) one strain carried TC resistance with a low transmission frequency that could produce colicin B. Genetic studies disclosed that the transmissible TC resistance factors, i.e., Rms105 (group b) and Rms104 (group c), were formed by recombination between Col B factor and nontransmissible TC-resistance (tet) determinant which appeared in E. coli P-38 mutants.

Animals↗

Plasmid-mediated sulfanilamide resistance.

Dihydropteroate synthetase (DHPS) is specified by a substrain of Escherichia coli K12, ML1410. This enzyme activity is inhibited by sulfanilamides (Sa) and is known to be heat-stable, i.e., an Sa-sensitive normal enzyme. Another DHPS activity specified by E. coli ML1410 carrying drug resistance plasmids is Sa-resistant but heat-sensitive, i.e., an Sa-resistant enzyme. Most plasmids encoding single Sa or double (Sa. Tc or Sa. Sm) (Tc, tetracycline; Sm, streptomycin) resistance mediate the formation of this type of DHPS. Therefore, E. coli carrying these plasmids becomes diploid for DHPS, i.e., an Sa-resistant and an Sa-sensitive normal enzyme. The biochemical mechanism of Sa resistance mediated by plasmids encoding triple (Cm.Sm.Sa; Tc.Sm.Sa) and quadruple (Cm.Tc.Sm.Sa) resistance is not due to the formation of an altered DHPS but probably due to the decrease in permeation of the drug into the cell. The evolutionary process of the formation of Sa-resistance determinants on plasmids is discussed based on the presence of two types of Sa resistance mechanism.

Bacteria↗

Antibacterial activity of apalcillin (PC-904) against gram-negative bacilli, especially ampicillin-, carbenicillin-, and gentamicin-resistant clinical isolates.

Apalcillin (PC-904) is active against carbenicillin- and ampicillin-resistant strains of gram-negative bacilli. Among Pseudomonas aeruginosa strains highly resistant to carbenicillin (>/=3,200 mug/ml), half of them were susceptible to PC-904 at a concentration of 50 to 1,600 mug/ml. The minimal inhibitory concentration of PC-904 against P. aeruginosa strains resistant to carbenicillin (400 to 1,600 mug/ml) ranged from 3.1 to 25 mug/ml. Ampicillin- and carbenicillin-resistant Enterobacteriaceae strains were similarly susceptible to PC-904. However, drug resistance to PC-904 was already apparent among some strains of P. aeruginosa, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, P. vulgaris, and P. morganii, recently isolated in Japan; i.e., 4, 35, 32, 4, 6, and 14% of strains isolated were resistant. PC-904 was more active, on the other hand, than ampicillin and carbenicillin against antibiotic-susceptible Enterobacteriaceae and also showed high activity against most species of Pseudomonadaceae, especially P. cepacia and P. aeruginosa. The minimum inhibitory concentrations of PC-904 were greatly affected by inoculum size when the organisms tested were strains producing large amounts of beta-lactamase.

Ampicillin↗

Streptomycin and Spectinomycin resistance mediated by plasmids.

Resistance to tetracycline (Tc), chloramphenicol (Cm), streptomycin (Sm), and sulfanilamide (Sa) was surveyed in clinical isolates of Escherichia coli and Shigella strains. Among the Sm Sa-resistant strains, the frequency of nonconjugative r (Sm Sa) plasmids was much higher than that of conjugative R plasmids encoding double resistance. The biochemical mechanism of Sm resistance mediated by the conjugative plasmids R(Tc Cm Sm Sa) and R(Cm Sm Sa) and about half the numbers of conjugative R(Tc Sm Sa) and R(Sm Sa) plasmids tested were found to be due to adenylylation of the drug. The remaining conjugative R(Tc Sm Sa) and R(Sm Sa) plasmids and all nonconjugative (Sm Sa) plasmids tested inactivated Sm by phosphorylation.

Conjugation, Genetic↗

New antipseudomonal penicillin, PC-904: affinity to penicillin-binding proteins and inhibition of the enzyme cross-linking peptidoglycan.

The mechanism of action of a new antipseudomonal penicillin, PC-904, was studied with respect to its binding affinities to penicillin-binding proteins (PBPs) and its inhibitory activities on cross-linking enzymes of peptidoglycan synthesis in vitro. PC-904 showed especially high affinity (compared with that of penicillin G) to Escherichia coli PBP-3. It also had high affinities to PBP-2 and -1Bs and low affinities to PBP-1A, -4, -5, and -6. Similar results were obtained with Pseudomonas aeruginosa, in which this antibiotic showed very high affinity (compared with that of penicillin G) to PBP-3, -1A (presumably corresponding to E. coli PBP-1Bs), and -2; there was especially high affinity to PBP-3 and much less affinity to PBP-1B (presumably corresponding to E. coli PBP-1A). These results are compatible with morphological observations that at concentrations near its minimal inhibitory concentration or less, this antibiotic induced the formation of filamentous cells of E. coli and P. aeruginosa. At higher concentrations or after prolonged incubation, it induced lysis of the cells. The remarkably high affinity of PC-904 to pseudomonal PBP-3, -1A, and -2 may partly explain the potent antipseudomonal activity of this antibiotic. In E. coli, the concentration of PC-904 required to inhibit the cross-linking reaction in enzymatic peptidoglycan synthesis, presumably carried out by PBP-1Bs, was as low as the inhibitory concentrations of penicillin G, ampicillin, and carbenicillin.

Ampicillin↗

Determination of the ID50 values of antibacterial agents in agar.

Microorganisms were plated on agar plates containing various concentrations of an antimicrobial drug and inhibition of growth was determined at each drug concentration. The ID50 value and the gradient of the line were calculated by the least square method. When 200 approximately 800 bacterial cells were inoculated on an agar plate, growth inhibition corresponded linearly with the log concentration of a drug within range of 5 approximately 95% inhibition. The ID50 value and the gradient obtained were reproducible and reliable using microorganisms at stationary phase of growth with all tested bacterial species and all tested antimicrobial agents. It was found that the ID50 values of drugs were more reproducible and may be more reliable than the MIC (minimum inhibitory concentration) values of the drugs.

Agar↗

Drug resistance of Escherichia coli isolated in Nepal.

We collected Escherichia coli strains from 59 Nepalese porters in 1971 and surveyed for their drug resistance. Drug-resistant E. coli strains were isolated from four porters. (TC. CM. SM. SA. APC.)-resistant strains were isolated from two porters and SA- or APC-resistant strains were isolated from each of the others. The R factors were demonstrated from the multiple-resistant E. coli strains.

Drug Resistance, Microbial↗

Biochemical properties of a penicillinase from Escherichia coli carrying Rms 298.

We obtained two R plasmids, i.e., Rms195 and Rms298, from a clinical isolate, E. coli GN5503. Penicillin beta-lactamase (PCase) was extracted from ML1410 Rms195+ and Rms298+, and was purified by chromatography. Rms195 PCase was identical to the type I PCase mediated by R-TEM, RI and Rms212. The isoelectric point of Rms298 PCase was 5.9 and its molecular weight was 21,000 +/- 1,000. The substrate profile and physiochemical properties indicate that Rms298 PCase belongs to the type IV PCase mediated by Rms139 isolated from Pseudomonas aeruginosa.

Cephalosporins↗

Temperature sensitive R plasmids isolated from Proteus strains.

Out of 32 R plasmids isolated from Proteus strains, 17 were found to be temperature sensitive with respect to inheritance in E. coli cells. They were fi- and classified into incompatibility group T or V. Cells carrying T group Rms273 plasmid were temperature sensitive with respect to growth and conjugal transfer in both E. coli and Proteus. The V group YOR-10 plasmid was stable in Proteus even at 42 C. However, the loss frequency of YOR-10 plasmid in E. coli reached 100% after 4 hr of incubation at 42 C, in spite of stable inheritance at 25 C. Conjugal transfer of the YOR-10 plasmid in E. coli was also strongly inhibited at 42 C. It has been concluded that instability of V group R plasmids in E. coli is due to their thermosensitive inheritance in the progeny cells at high temperatures.

Escherichia coli↗

Antibacterial and inducer activities for tetracycline resistance by its derivates and analogues.

Antibacterial and inducer activities of thirteen TC derivates were investigated for tetracycline (TC) resistance in staphylococcus aureus. Four compounds of the TC derivatives were not able to induce the resistance to TC in Staphylococcus aureus MS3937 rms7 (TC) + harboring an inducible TC resistance determinant located on a plasmid. The 12a-hydroxy position seems to be essential for the inducer activity.

Bacterial Proteins↗

Inducible high resistance to colistin in Proteus strains.

Wild-type Proteus strains are usually resistant to colistin. We have observed an increase in the level of colistin resistance in four clinical isolates of P. vulgaris, P. morganii, and P. rettgeri by prior treatment with subinhibitory concentrations of the drug. The enhanced resistance in these strains was lost after cultivating them in colistin-free medium, indicating that the enhanced resistance in the Proteus group is inducible.

Colistin↗

Nonconjugative plasmids encoding sulfanilamide resistance.

Nonconjugative plasmids encoding sulfanilamide (Sa) resistance were demonstrated at a high frequency in Shigella and Escherichia coli strains resistant to sulfanilamide. These Sa plasmids were all compatible with the standard plasmids used in compatibility testing. The sizes of seven Sa plasmids were measured by electron microscopy and ranged from 1.79 to 2.08 mum, corresponding to 3.5 to 3.9 megadaltons.

Conjugation, Genetic↗