Tissue-nonspecific alkaline phosphatase (TNSALP) in serum comprises liver alkaline phosphatase (liver-ALP) and bone alkaline phosphatase (bone-ALP). Liver-ALP is a marker of liver disease; thus a specific method for its measurement would be useful. Measurement of ALP by electrophoresis is difficult, although all of the isozymes can be assessed simultaneously. Total ALP can also be measured by automated analyzer, but it is difficult to determine the cause of a high ALP value because bone-, intestine-, placenta-, and tumor-ALP are measured together. Thus, anti-TNSALP monoclonal antibodies that can resolve these problems are needed. Here we have generated an anti-TNSALP monoclonal antibody, 3-29-3R. This clone has specificity to liver-ALP rather than to bone-ALP. In electrophoresis, 3-29-3R reacted with TNSALP and shifted the bands. The use of 3-29-3R enabled easy interpretation of the results. Furthermore, we tested 3-29-3R by developing an immunocapture enzymatic assay (IEA). Preliminary results of the IEA show that this method is effective for measurement of liver-ALP. Thus, the monoclonal antibody that we have established may be a useful tool for clinical diagnosis.
When dl -isoleucine was added to an ammonium nitrate-maltose medium during cultivation of Streptomyces sp. 732, quinomycin B formation was selectively enhanced (from 3 to 70% of the quinomycin mixture) and quinomycin C synthesis was inhibited completely. In addition, two new quinomycins, designated quinomycins D and E, were produced in the presence of isoleucine. These compounds were found to contain N -methylalloisoleucine. In experiments with isoleucine enantiomorphs, it was determined that the order of effectiveness for quinomycin B synthesis was dl -isoleucine > d -isoleucine > l -isoleucine. The extent to which quinomycin B synthesis is enhanced depends upon the concentration and the time of addition of isoleucine to the medium. The effect of dl -isoleucine was reduced to some extent by the addition of l -valine. It is conceivable that amino acids which are precursors of the N -methylamino acids in quinomycin can regulate quinomycin formation.
Fifteen strains which showed a specific activity to acid-fast bacteria only in streak plate method, were tested using several kinds of culture media on their antibiotic potency in the culture filtrates, and studied further on the nature of antibiotic substances from the point of their own antibiotic spectra, active pH and diffusibility.
Serum band 5 tartrate-resistant acid phosphatase (TRACP 5; EC 3.1.3.2) is a glycoprotein that exists as two very similar isoforms, TRACP 5a and TRACP 5b. The similarity of these two isoforms has made it difficult to establish monoclonal antibodies specific for either isoform. We report here the development of a monoclonal antibody with high specificity for TRACP 5b. We prepared TRACP 5b antigens from four sources: TRACP 5b purified from human bone, recombinant TRACP 5 from Escherichia coli, recombinant TRACP 5 from insect cells, and a synthetic TRACP 5b peptide. Thirty-seven mice were each immunized with 1 of the 4 different TRACP antigens to generate 473 antibody-producing clones. Three of these clones, Trk27, Trk49, and Trk62, reacted with TRACP 5b. These three clones were all established from mice exposed to native bone TRACP 5b antigen. In fact, none of the other antigens were able to generate anti-TRACP 5b monoclonal antibodies in mice. Furthermore, Trk62 interacted more strongly with TRACP 5b than with TRACP 5a. These results suggested that although recombinant proteins can be effective antigens, the native TRACP 5 protein might be more effective at generating monoclonal antibodies of greater specificity due to its more faithful representation of the native three-dimensional structure of the protein.
1) Five kinds of basic antagonistic substances, which resembled streptothricin in nature, were obtained in a form of hydrochloride, crystal-line helianthate and reineckate, and the identification between these sub-stances was performed. 2) All of these substances had rather lower potencies than strepto-mycin, in a dilution titer against E. coli, while the substances No. 259 and No. 120 (Seki) were higher inhibitory against Staphylococcus (Terashima strain) than streptomycin. 3) Six substances, including streptomycin, were arranged in the order of increasing toxicities as follows: -36, streptomycin, 120 (Seki), 24, 39, and 259. The substance No. 36 was able to be injected both, intravenously and intramuscularly into mice in an amount of 20mg without showing any toxic signs. In this respect, the substance No. 36 has advantages over streptomycin. 4) The decomposition points of helianthates or reineckates of 4 streptothricin-Tike substances were found to be in the vicinities of that of streptothricin except No. 36 (Roseomycin). In reference to the other biological and chemical properties, however, the 5 substances seem quite different from streptomycin and from each other. 5) Maltol and Sakaguchi reactions were found to be positive in streptomycin alone, while glucosamine and other sugar reactions were positive in both streptomycin and streptothricin-like 5 substances. The chemical reactions positive in streptothricin group only, were not found yet. 6) A strain of E. coli, which was made resistant to the hydrochlorides of 4 substances (36, 20, 259, and 120) respectively was resistant to each of 5 substances and somewhat less resistant to streptomycin too, while the same strain of E. coli, which was made resistant to streptomycin, was not resistant to each of 5 substances. In this respect it seems very probable, that each of 5 substances resembles closely in their mechanisms of the anti-biotic functions.
To examine the influence of sex hormones on the appearance of overt diabetes in NOD mice, oophorectomy and orchiectomy were performed. Castrated males showed a high incidence of diabetes. The time course of onset was similar to that of the intact females. On the other hand, castrated females showed a intact-male pattern of onset with a low incidence. Clinical determinations demonstrated a close similarity between spontaneous and castration-induced diabetes. These data suggest that sex hormones modulate the expression of overt diabetes in NOD mouse, i. e. androgens have at least a suppressive effect.
