Biomedical subjects
Publications and source records attributed to M Tai.
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The monomer-single polymer model of G.A. Gilbert (Disc. Faraday Soc. 20 (1955) 68) for moving boundary sedimentation has been used by Payens and colleagues to explain the observed results for bovine caseins, and by Harrington and colleagues to explain the observed results for myosin fibrils. Electron microscope pictures of Buchheim and Schmidt have subsequently revealed micellar beta-casein in the form of slightly elongated or spherical particles having a bimodal size distribution, but with a broad range of particle sizes, at concentrations not too far above the critical micelle concentration. The equilibrium properties of a broadly distributed micellar system can be fitted by the shell model developed by one of us, and in the present article, the shell model is extended to predict the moving boundary sedimentation behavior of such a system. The observed sedimentation patterns, as well as the critical concentration predictions of the monomer-single polymer Gilbert sedimentation model, are satisfactorily described with the present model, based on a continuous distribution of intermediates between monomers and the largest possible spherical micelles. For one example considered, the predicted frequency distribution of molecular weight is in qualitative agreement with the frequency distribution of particle volume found by Buchheim and Schmidt.
Two samples of highly purified bovine beta-casein supplied to us by Dr. T.A.J. Payens of the National Institute for Dairy Research, Ede, The Netherlands, were studied over a range of concentration from just below the critical micelle concentration (CMC) to 0.46%, in 0.2 ionic strength phosphate buffer (pH 7.0), at 20 and 25 degrees C. The relaxation process studied by pressure jump using a 90 degree scattered light detector was also confirmed by the temperature-jump method. In the pressure-jump experiments, the process could be separated into two general time domains, with an approximate ratio of 10-25:1, a behavior reminiscent of that found for synthetic micellar systems. The faster relaxation process was still exhibited below the CMC, however. The concentration dependence of the faster relaxation time agreed very satisfactorily with predictions from the micelle model described in the companion paper.
Ultracentrifugal analysis of ribosomal purity is complicated by the rapid reequilibration of ribosomes with their subunits, and this is further enhanced by the effects of hydrostatic pressure. Fixation of the ribosome system prior to ultracentrifugal analysis supposedly freezes the reequilibration, and thus tends to obviate these difficulties. However, no redistribution of the ribosome-subunit population must be allowed to occur during fixation. Thus, it is necessary that fixation be extremely rapid compared to the ribosome-subunit reequilibration, in order to avoid errors in analysis. It was the purpose of this investigation to make a direct experimental comparison of the rates of these two processes, fixation and ribosome-subunit reequilibration, using the stopped-flow technique with a light-scattering detector, under a variety of buffer environment readjustments. The following findings resulted from this study: (1) Fixation in Tris buffers could not be followed by light scattering because of interaction of glutaraldehyde with Tris, leading to continuous production of high molecular weight contaminants. (2) Polymerization of glutaraldehyde leads to increased ultraviolet absorption, which must not be confused with scattering changes. (3) Undialyzed ribosome solutions prepared by dissolving stock suspensions stored at high levels of magnesium and univalent electrolyte into a known standard buffer produced solutions having free Mg2+ levels lower than those of original buffer, complicating kinetic observations and threatening ribosome stability. (4) Addition of malonic acid as a buffer for Mg2+ largely eliminated this problem. (5) The kinetics of glutaraldehyde were measured quantitatively by studying the perturbation of ribosome association and dissociation kinetics produced during shifts of Mg2+ levels. (6) At fixative levels not producing coagulation, fixation kinetics can be competitive with those of association-dissociation. (7) Mass action effects like those of dilution can be caused by fixation, and can give rise to excess subunits, which may be mistaken for loose couples in original ribosome preparations.
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A method has been developed for quantitative determination of the early appearing mouse paw swelling activity of pertussis vaccine. Mice were inoculated with the vaccine into the left hind paws. The difference in the thicknesses between the inoculated and the uninoculated paws was taken as the swelling response in 16 hr after inoculation. It was transformed into logarithm as a response metameter. A linear log dose-log response regression line was obtained over a relatively wide range of doses of the vaccine. No deviation from parallelism was significant among the regression lines of different vaccines tested. The swelling activities relative to that of a reference vaccine of diphtheria-pertussis-tetanus combined (DPT) vaccines recently produced in Japan were determined. The relative activities ranged from 0.6 to 2.5, being twice the mean width of the 95% confidence interval of the relative activity. Further experiments showed that the activity significantly correlated with endotoxin and histamine-sensitizing factor (HSF) but not with lymphocytosis-promoting factor (LPF). In addition, the multiple regression analysis of such relationship suggested that the swelling activity is accounted for by the combined action of endotoxin and HSF. A model experiment with a mixture of the two toxins supported this conclusion.
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The activities of T-1982 (sodium 7 beta-[(2R, 3S)-2-(4-ethyl-2, 3-dioxo-1-piperazine-carboxamido)-3-hydroxybutanamido]-7 alpha-methoxy-3-[(1-methyl-1H-tetrazol-5-yl)thiomethyl]-3-cephem-4-carboxylate) against various gram-positive and gram-negative bacteria were compare with those of cefmetazole, cefoxitin, cefazolin, and cefoperazone. T-1982 was active against both gram-positive and gram-negative bacteria, including genera resistant to the other cephalosporins. T-1982 exhibited greater activity than did cefmetazole, cefoxitin, cefazolin, or cefoperazone against Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, and Serratia marcescens and was also highly active against Bacteroides fragilis. T-1982 was as stable to various beta-lactamases as were cefmetazole and cefoxitin. The therapeutic activities of T-1982 in mice experimentally infected with various gram-negative bacteria were superior to those of cefmetazole, cefoxitin, cefazolin, and cefoperazone.
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The concentration of hepatic nuclear T3 receptors was measured in isolated nuclei from athyreotic mice bearing thyrotrophic tumors and intact rats with Walker 256 carcinoma. Receptor concentration was reduced in all tumor-bearing animals. The mean receptor capacity of the Walker tumor-bearing rats [0.31 +/- 0.05 (SEM) ng/mg DNA] was significantly decreased from simultaneously assayed controls (0.47 +/- 0.04 ng/mg DNA; P less than 0.01). No change in the apparent equilibrium association constant was observed. In individual rats, the magnitude of the decrease in nuclear T3 receptor concentration was highly correlated with the decrease in tumor-free body weight. Additional studies showed that the decrease in nuclear receptors was not due to delayed equilibration of added T3 with nuclear sites in vitro or to an increase in endogenous hepatic T3 concentration. The plasma concentration of total and free T4 and T3 was decreased in tumor-bearing rats. Plasma TSH concentration, however, remained unchanged. Thus, these transplantable neoplasms seem to be associated with decreased hepatic nuclear receptors and low concentrations of plasma thyroid hormones. The unchanged plasma TSH suggests that the animals remained euthyroid.
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.
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T-1220, sodium 6-[d-(-)-alpha-(4-ethyl-2,3-dioxo-1-piperazinylcarbonyl-amino)- alpha-phenylacetamido] penicillanate, is a new semisynthetic penicillin derivative that possesses a broad spectrum of in vitro antibacterial activity against gram-positive and gram-negative bacteria. T-1220 is more effective than carbenicillin (CB-PC) against Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus species, and Serratia marcescens. Addition of human serum to culture media did not significantly alter the antibacterial activity of T-1220. Greater bactericidal activity toward various strains of gram-negative bacteria was demonstrated with T-1220 than with CB-PC. T-1220, like penicillin G, was hydrolyzed by penicillinase, but was sable to a type IV penicillinase produced by P. aeruginosa strains. In vivo antibacterial activities of T-1220, ampicillin (AB-PC), and CB-PC were compared, using systemic infections of mice with P. aeruginosa, K. pneumoniae, and Escherichia coli. The 50% effective doses (milligrams per kilogram) of T-1220 were consistently lower than those of AB-PC and CB-PC.
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