The science reports of the research institutes, Tohoku University. Ser. C, Medicine. Tohoku Daigaku最新文献

A stimulatory factor of DNA-dependent RNA polymerase B (nucleosidetriphosphate: RNA nucleotidyltransferase, EC 2.7.7.6) in nonhistone proteins was partially purified from rat liver nuclei on a column of daunomycin-CH Sepharose 4B and of phosphocellulose. In the process of purification, the stimulatory factor was separated from the main fraction of nuclear protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37). This factor enhanced specifically the activity of RNA polymerase B on rat liver DNA as template and did not affect RNA polymerase A and Escherichia coli RNA polymerase at all. The polynucleotide elongation rate was increased by the addition of this factor.

The effects of cytochalasin B (CB) and colchicine on the motility and growth of cultured Yoshida sarcoma cells are studied by cinematographic methods. CB was found to reduce the average locomotory rate of motile Yoshida sarcoma cells and to enhance the frequency of non-motile cells. On the other hand, colchicine enhanced only the frequency of non-motile cells and did not affect the locomotory rate of motile cells. CB inhibited the growth of Yoshida sarcoma cells at the concentration range of 0.6 to 5 microgram/ml.

A 44-year-old male was hospitalized, because of the presence of abnormal shadows on chest x-ray film. Diagnosis of pulmonary alveolar proteinosis was established by examination of a specimen obtained by transbronchial biopsy. Therapeutic bronchopulmonary lavage was performed twice. As a result, there was an improvement of radiological findings on chest x-ray film. Then a biochemical study was carried out with pooled lavage fluids. Cellular debris were removed by centrifugation. Lipids were extracted with a 2:1 mixture of chloroform and methanol. Individual phospholipids were identified by column chromatography and thin layer chromatography. Fatty acids were identified by gas chromatography. It was found that phosphatidyl choline (lecithine) was the major component of phospholipids. On the other hand, myristic acid of the molecule of phosphatidyl choline was found to constitute 19.6% of the total fatty acids. This patient lives a normal life as of Dec. 1980.

Biological effects of animal lung proteoglycans on functions of pulmonary alveolar macrophages from the same animal species were evaluated by quantitative and qualitative NBT tests. It was found that fractions of lung proteoglycans modulated the rate of NBT reduction in pulmonary macrophages under selected conditions.

1. Pleural fluid contained protein-bound hyaluronic acid, protein-bound chondroitin sulfate, hyaluronic acid, chondroitin sulfate, undersulfated chondroitin sulfate and dermatan sulfate. The composition of acid glycosaminoglycans in pleural fluid seems to reflect the rate of biosynthesis and degradation of these polysaccharides at some sites which are closely related to the pleural cavity. 2. A possibility was suggested that hyaluronic acid was synthesized in pleural tissue and was excreted shortly thereafter into the surroundings, as evidenced by experiments with rabbit pleural tissue. 3. In human, hyaluronic acid, chondroitin sulfate, dermatan sulfate and heparan sulfate were found in thickened pleurae caused by lung cancer, in those caused by asbestosis and also in tumor tissues of pleural mesothelioma. The molecular size of hyaluronic acid from pleural mesothelioma was found to be larger than that from human unbilical cord. 4. Quantification and histochemical study of acid glycosaminoglycans demonstrated that the quantity of hyaluronic acid in tissue specimens of mesothelioma by far exceeded that in non-mesothelioma cases (statistically significant). 5. Thus a possibility was suggested that histochemical investigation together with microquantitation of hyaluronic acid in pleural tissue may prove to be an efficient means of differential diagnosis of pleural mesothelioma. 6. Definite conclusion on the relationship between the fluctuation with time in quantity of acid glycosaminoglycans of the effusions and etiology of pleurisy awaits further investigations.

1. The susceptibility of M. pneumoniae to antibiotics can be determined by the microtiter method. The adequate technique requires that the final volume of broth medium in a well is 0.2 ml and that the dilution is made after the parent solution of antibiotic in the test tube is dropped into a well every fifth wells. 2. M. pneumoniae was cultured on agar media containing two-fold concentrations of macrolide and analogous antibiotics, and the following results were obtained. 1) The growth of eight strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of thet strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of thet strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of the colony grown as an average of 0.5 to eight on agar media containing erythromycin in four strains was 0.1 to 1.6 micrograms/ml in some colonies, and 400 to 800 micrograms/ml in most colonies. The results disclosed that the broth culture contains a small number of mycoplasma cells with a definite, high degree of resistance to the antibiotics, but no cells with intermediate degrees of resistance. 3) The FH strain was made resistant to erythromycin, oleandomycin, midecamycin, acetylspiramycin, leucomycin, josamycin, tylosin, lincomycin, or clindamycin by subculture in broth medium from the colony grown at the highest concentrations of each of the antibiotics in agar media. The degree of the resistance developed was 16 to 128,000 in the MIC radio and showed high values of MIC in most strains. The resistance developed was not lost by subculturing the resistant strain in broth medium without antibiotic. 4) The FH strain made resistant to the antibiotics had cross resistance to other macrolides. Strains resistant to some of the antibiotics had cross resistance to lincomycin and clindamycin, and strains resistant to others did not. Some strains made resistant to macrolides with cross resistance to lincomycin and clindaymycin and strains made resistant to lincomycin or clindamycin had no cross resistance to vernamycin B alpha, while all the resistant strains without cross resistance to lincomycin and clindamycin had cross resistance to vernamycin B alpha. No strain had cross resistance to vernamycin A...

1. Glucosamine 6-phosphate (GlcN-6-P) isomerase of rat kidney was resistant to heating at 50--55 degrees in crude extract but not after several purification steps. GlcN-6-P and N-acetylglucosamine 6-phosphate were found to stabilize the isomerase under these conditions. They also protected the enzyme from tryptic digestion, but only GlcN-6-P was effective against inactivation by p-chloromercuribenzoate. 2. When GlcN-6P isomerase was purified from fresh kidney and kidney stored at -20 degrees, separately and under GlcN-6-P, the two preparations were different in elution profile from a hydroxyapatite column. It was subsequently found that storage of crude extract at -20 degrees resulted in molecular alterations of the enzyme. Prolonged purification appeared to affect the enzyme similarly. The molecular alterations, however, were suppressed if the extract was stored at -70 degrees. 3. These findings have been utilized to develop a procedure, which enables us to purify rat kidney GlcN-6-P isomerase without any molecular alteration and in good yield.

Low concentration (0.1--1 mM) of ascorbate and erythorbate (isoascorbate) caused lipid peroxidation and lysosome labilization ("cofactor" action). In addition, they acted additively on microsomal NADPH oxidase-induced lipid peroxidation at the low concentration. The "cofactor" action, however, was dependent reciprocally on the density of lysosomes; the more dilute was the lysosomal fraction, the more susceptible the lysosomes were. On the other hand, ascorbate and erythorbate at concentration more than 1 mM inhibited microsomal NADPH oxidase-induced lipid peroxidation and lysosome labilization. Their antioxidant effect was revealed to be clear especially when the "cofactor" action was eliminated by such a basic protein as protamine. Considering that the "cofactor" action was observed only at the lower density of lysosomes and might be inhibited by physiologically occurring basic proteins, ascorbate and erythorbate may mostly act as antioxidant on lysosomes in vivo. Ascorbate- or erythorbate- induced lysosome labilization was certified to be mediated by lipid peroxidation.