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Biomedical subjects

K Uehara

Publications and source records attributed to K Uehara.

At least 145 records · Page 8Linked to original sources

[Bacteriological and clinical studies of cefoxitin with special reference to anaerobic infections in the patients of abdominal surgery (author's transl)].

Both bacteriological and clinical studies of the effectiveness of cefoxitin (CFX) in the treatment of infections associated with abdominal surgery have been carried out at Tokyo Metropolitan Toshima Hospital from September 1979 through August 1980. The results of these studies are summarized in the following: 1. The clinical isolates from the 29 surgical patients were studied and anaerobes were found in 16 patients (55% of the patients). B. fragilis was found in 11 of the 16 patients (69%) from whom anaerobes were isolated. In all of these patients, aerobes were also isolated--most frequently E. coli or K. pneumoniae. 2. The in vitro antibacterial activity of cefoxitin (CFX) against 83 clinical isolates was compared to that of cephalothin (CET), cefazolin (CEZ) and carbenicillin (CBPC). The activity of CFX against Gram-positive bacteria was generally slightly inferior to that of the other 3 antibiotics. Among the Gram-negative aerobic organisms, CEZ was the most active against E. coli and CFX and CEZ against K. pneumoniae. However, CFX, with MIC's of 0.78-12.5 micrograms/ml, showed the greatest activity against B. fragilis, followed by CBPC, CEZ and CET, in order of decreasing activity. 3. CFX was administered in a 1-hour drip infusion to 3 patients following abdominal surgery, and concentrations of CFX in the serum and the exudate were measured. Peak serum concentrations were obtained at the end of the infusion, with a mean peak level of 97.93 micrograms/ml. Peak concentrations in the exudate were observed 30 to 60 minutes later and varied from 21.10 to 56.25 micrograms/ml. 4. Of the 20 patients administered of CFX, complete clinical and bacteriological data of anaerobic infections were available in 8 patients. The clinical evaluation was 'good' in 7 patients and 'fair' in 1. The bacteriological evaluation was 'eradicated' in 5 patients and 'decreased' in 3. As for side effects, elevations of S-GOT and S-GPT were observed in 4 of the 20 patients received CFX, but these abnormalities might also be attributable to other factors such as underlying disease, surgical intervention etc. No other side effects were found in these patients.

Abdomen↗

Evidence for phase separation in the membrane of an osmotically stabilized fatty acid auxotroph of E. coli and its biological significance.

1. An unsaturated fatty acid auxotroph of Escherichia coli accumulated a high content of saturated fatty acids in its membrane when it was cultured under osmotically stabilized conditions. The physicochemical properties of the phospholipid extracts and of the membrane fraction from the cells were investigated by means of proton magnetic resonance, infrared spectroscopy and differential scanning calorimetry. 2. Physicochemical studies indicate that the phospholipid bilayers in the membranes exhibit at least two phase transitions, a minor one at approx. 19 degrees C and a major one at approx. 43 degrees C. Between the two temperatures, gel and liquid crystalline domains co-exist. Moreover, even in the gel state, phospholipids seem to segregated into domains containing different proportions of unsaturated fatty acids. 3. The Arrhenius plot of beta-galactoside transport rates is biphasic. The inflection point is at 22 degrees C. This means that the appearance of the fluid region in the bilayer at approx. 19 degrees C is important in the activation of membrane transport.

Biological Transport↗

Studies on D-tetrose metabolism. Crystallization and properties of D-erythrulose reductase from chicken liver.

D-Erythrulose reductase from chicken liver has been purified to homogeneity as judged by acrylamide gel electrophoresis and ultracentrifugation. The overall purification of the enzyme was 164-fold from a crude extract. The enzyme was crystallized from ammonium sulfate solution at pH 7.0 to give hexagonal plates. The molecular weight determined by sedimentation equilibrium analysis was 94,600 and that by SDS-polyacrylamide gel electrophoresis was 22,400, which suggests a tetrameric structure for the native enzyme. The enzyme was found to contain up to 3 molecules of NADP+ per enzyme; this high amount of NADP+ resulted in a higher absorption at 260 nm than at 280 nm. The extinction coefficient of the enzyme at 290 nm was found to be 4.0. The contents of various amino acids were very similar to those of the beef liver enzyme formerly crystallized in our laboratory. The isoelectric point of the enzyme determined by Ampholine isoelectric focusing was pH 6.43. The enzyme was shown to catalyze the reduction of D-erythrulose to D-threitol with the concomitant oxidation of NAD(P)H to NAD(P)+, and was highly specific to D-erythrulose with an apparent Km of 0.38 mM. NADH was less effective than NADPH and the Km's for NADH and NADPH were 67 micrometers and 7.9 micrometers, respectively. D-Threitol was slightly oxidized by the enzyme with either NADP+ or NAD+ as a cofactor at pH's 7.5 and 9.0.

Amino Acids↗

Effect of NADP+ and its analogs on the Rose Bengal-sensitized photoinactivation of D-erythrulose reductase from beef liver.

Upon addition of NADP+, the rose bengal-sensitized photoinactivation of D-erythrulose reductase from beef liver is prevented to a remarkable extent. Adenosine 2',5'-diphosphate (2',5'-ADP) also has a protective effect, but to a lesser extent. On the other hand, 2'-AMP markedly enhances the photoinactivation. Other nucleotides which have no 2'-phosphoryl group, such as NAD+, 3'-AMP, 5'-AMP, ADP, and NMN, are ineffective. Further, only 2'-AMP derivatives (NADP+, 2',5'-ADP, and 2'-AMP) among these nucleotides were found to be potent competitive inhibitors of the enzyme with small Ki's (6--13 muM). Photooxidation of some methionine residues in the enzyme is prevented by the addition of NADP+ and accelerated in the presence of 2'-AMP. Photooxidation products(s) of 2'-AMP derivatives have no effect upon the enzymatic activity. Although NADP+ and 2'-AMP induce detectable conformational changes of the enzyme, the changes are not characteristic to the compounds. Based on these observations, we present a possible action mechanism of 2'-AMP derivatives on the photoinactivation of D-erythrulose reductase.

Adenosine Monophosphate↗

Phase transitions of phospholipid bilayers from an unsaturated fatty acid auxotroph of Escherichia coli.

Total phospholipids were extracted from cells of temperature sensitive unsaturated fatty acid auxotrophs of Escherichia coli (K-12 UFAts) grown at 28degrees C (PL28), and at 42degrees C in the presence of 2% KCl as an osmotic stabilizer (PL42 (KCl)). From the analysis of fatty acids, it was shown that the content of unsaturated fatty acids of PL42 (KCl) is only 9% of the total fatty acids, while that of PL28 is 54%. The thermal phase transitions of the bilayers prepared from the phospholipid fractions were studied by proton magnetic resonance. The line widths of the methylene signals and the sums of the methylene and methyl signal intensities were plotted against reciprocal values of absolute temperature 1/T or temperature itself. From the plots phase transitions were detected at about 19degrees C for PL28 and at 43degrees C for PL42 (KCl). In spite of its complex composition of fatty acids a highly cooperative transition was observed in the case of PL42 (KCl). It was also suggested that the phospholipids bilayers in the biomembranes of this strain at the growth temperature (42 degrees C) are in the state where the gel and liquid crystalline phases coexist.

Cell Membrane↗