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961.
Antibody to cilia-associated respiratory (CAR) bacillus was detected by the indirect immunofluorescence assay (IFA) technique using tracheal sections of infected mice as antigen in serum samples collected from rats infected naturally and experimentally. Nine of 23 cases of natural infection were positive in IFA antibody, with titres ranging from 1:10 to 1:80, and all these antibody-positive cases were also histologically positive. The remaining 14 cases were negative in both IFA antibody and histological diagnosis, even though some of them were infected with Sendai virus and Mycoplasma pulmonis. In the experimental infection, serum samples collected from 18 rats on days 4, 7, 14, 21, 28 and 56 post-inoculation (PI) (three rats for each point) and examined for IFA antibody revealed that seroconversion occurred in one rat on day 14 PI and in three rats on day 21 PI. Antibody titres of 1:80 to 1:160 remained to the termination of the experiment. The IFA technique was useful for the diagnosis of CAR bacillus infection except in the early stage of the infection. 相似文献
962.
H Ikezawa M Matsushita M Tomita R Taguchi 《Archives of biochemistry and biophysics》1986,249(2):588-595
Some divalent metal ions were examined for their effects on sphingomyelinase activity of Bacillus cereus. The enzyme activity toward mixed micelles of sphingomyelin and Triton X-100 proved to be stimulated by Co2+ and Mn2+, as well as by Mg2+. Km's for Co2+ and Mn2+ were 7.4 and 1.7 microM, respectively, being smaller than the Km for Mg2+ (38 microM). Sr2+ proved to be a competitive inhibitor against Mg2+, with a Ki value of 1 mM. Zn2+ completely abolished the enzyme activity at concentrations above 0.5 mM. The concentration of Zn2+ causing 50% inhibition of the enzyme activity was 2.5 microM. Inhibition by Zn2+ was not restored by increasing concentrations of Mg2+ when the concentration of Zn2+ was above 10 microM. Ba2+ was without effect. When sphingomyelinase was incubated with unsealed ghosts of bovine erythrocytes at 37 degrees C, the enzyme was significantly adsorbed onto the membrane in the presence of Mn2+, Co2+, Sr2+ or Ba2+. Incubation with intact or Pronase-treated erythrocytes caused enzyme adsorption only in the presence of Mn2+. In the course of incubation, the enzyme was first adsorbed on the membranes of intact bovine erythrocytes in the presence of Mn2+; then sphingomyelin breakdown proceeded with ensuing desorption of adsorbed enzyme. Hot-cold hemolysis occurred in parallel with sphingomyelin breakdown. In this case, the hydrolysis of membranous sphingomyelin as well as the initial enzyme adsorption took place in the following order: unsealed ghosts greater than Pronase-treated erythrocytes greater than intact erythrocytes. 相似文献
963.
Reconstitution of pyrroloquinoline quinone-dependent D-glucose oxidase respiratory chain of Escherichia coli with cytochrome o oxidase. 总被引:1,自引:1,他引:0 下载免费PDF全文
D-Glucose dehydrogenase is a pyrroloquinoline quinone-dependent primary dehydrogenase linked to the respiratory chain of a wide variety of bacteria. The enzyme exists in the membranes of Escherichia coli, mainly as an apoenzyme which can be activated by the addition of pyrroloquinoline quinone and magnesium. Thus, membrane vesicles of E. coli can oxidize D-glucose to gluconate and generate an electrochemical proton gradient in the presence of pyrroloquinoline quinone. The D-glucose oxidase-respiratory chain was reconstituted into proteoliposomes, which consisted of two proteins purified from E. coli membranes, D-glucose dehydrogenase and cytochrome o oxidase, and E. coli phospholipids containing ubiquinone 8. The electron transfer rate during D-glucose oxidation and the membrane potential generation in the reconstituted proteoliposomes were almost the same as those observed in the membrane vesicles when pyrroloquinoline quinone was added. The results demonstrate that the quinoprotein, D-glucose dehydrogenase, can reduce ubiquinone 8 directly within phospholipid bilayer and that the D-glucose oxidase system of E. coli has a relatively simple respiratory chain consisting of primary dehydrogenase, ubiquinone 8, and a terminal oxidase. 相似文献
964.
Purification and Properties of beta-N-Acetylhexosaminidase from Mucor fragilis Grown in Bovine Blood
Yamamoto K Tsuji Y Matsushita S Kumagai H Tochikura T 《Applied and environmental microbiology》1986,51(5):1019-1023
Mucor fragilis grown on bovine blood powder as the sole carbon source abundantly produced beta-N-acetylhexosaminidase. The enzyme activity was several times higher than that of a culture obtained with glucose medium. The enzyme had two different molecular weight forms. The high-molecular-weight form had somewhat higher beta-N-acetylgalactosaminidase activity than the lower-molecular-weight enzyme which had beta-N-acetylgalactosaminidase activity equivalent to about 40% of its beta-N-acetylglucosaminidase activity. Bovine blood seemed to induce both enzymes, but N-acetylamino sugars specifically induced the low-molecular-weight form. N-Acetylgalactosamine had an especially marked effect on activity. The low-molecular-weight form of enzyme was purified from the culture filtrate by fractionation with ammonium sulfate and various column chromatographies. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis. The optimum pH was 4.0 to 5.0 for beta-N-acetylglucosaminidase activity and 5.5 to 6.5 for beta-N-acetylgalactosaminidase activity. The enzyme hydrolyzed natural substrates such as di-N-acetylchitobiose, tri-N-acetylchitotriose, and a glycopeptide obtained by modification of fetuin. 相似文献
965.
A cell-binding, immunoglobulin-like protein from human plasma. I. Isolation and subunit structure 总被引:1,自引:0,他引:1
A protein having Mr of more than 900K (900,000) was isolated by chromatography on a column of Sepharose 4B coupled with solubilized elastin, followed by sucrose density gradient centrifugation. The protein is composed of several disulfide-linked subunits. SDS-polyacrylamide gel electrophoresis and immunoblotting indicated that two of the subunits are identical with the heavy chains of IgM and IgG, and one is immunologically related to the heavy chain of IgA. The results suggested that this protein is a new protein that belongs to the category of, or is closely related to, the immunoglobulins. As described in the succeeding paper, it has cell-binding activity (Fukamizu et al. (1986) J. Biochem. 100, 843-848). We tentatively refer to this protein as "cell-binding immunoglobulin-like protein (CIP)." 相似文献
966.
mRNA was prepared from autopsy liver samples from a homozygote for α1-antitrypsin deficiency (PiZZ) and from a normal (PiMM) subject. Both preparation gave equivalent synthesis of α1-antitrypsin in a wheat germ cell-free system. This suggests that the deficiency of plasma α1-antitrypsin associated with the Z variant is due to a failure of processing and secretion of the protein rather than of its synthesis. It is likely that it is the resultant intracellular accumulation of the Z protein rather than a deficiency of protease inhibitor that is the primary cause of the liver pathology associated with this variant. 相似文献
967.
Tests for Deoxyribonucleic Acid Synthesis in Toluenized Bacillus subtilis Cells 总被引:2,自引:1,他引:1 下载免费PDF全文
Several tests were devised to further characterize deoxyribonucleic acid (DNA) synthesis in toluenized Bacillus subtilis cells. Vigorous agitation of toluenized cells (localization test) demonstrated that the DNA replication is exclusively a cell-associated process. A DNA "repair" condition was also applied to toluenized cells and shown to be distinct from DNA replication in its DNA polymerase I dependency and its ability to synthesize DNA on template which is either cell associated or free, outside the cell. This repair condition was used in conjunction with the localization test to demonstrate the penetration of deoxyribonuclease I and possibly DNA polymerase I into toluenized cells. Therefore, we suggest that the localization test can be used to test the penetration of proteins into toluenized cells for both the DNA repair and replication processes. 相似文献
968.
Permanent loss of chromosome initiation in toluene-treated Bacillus subtilis cells. 总被引:2,自引:1,他引:1 下载免费PDF全文
Initiation of deoxyribonucleic acid replication is absent in Bacillus subtilis cells made permeable by toluene. The absence of initiation may be (i) a temporary removal of toluene, or (ii) irreversibly lost due to damage by toluene treatment to a cellular structure or a process required for chromosome initiation. Washed cells, previously treated with toluene and subsequently washed free of detectable amounts of toluene, have the same characteristics of toluene-treated cells in which toluene is not removed. The continued absence of initiation in the washed cells indicates a permanent loss of initiation in cells treated with toluene. Protein synthesis is also inhibited irreversibly by toluene treatment, indicating damage to translation as a possible mechanism for loss of initiation of toluene-treated cells. 相似文献
969.
Electron Microscopy of the Combination of Antibodies with Flagellar Antigen and with a Pyocine 下载免费PDF全文
Micrographs are presented of antibodies in combination with flagella of Salmonella typhi and with a phage-bound pyocine Rmc, which is supposed to be the tail of a defective bacteriophage from Pseudomonas aeruginosa. The pyocine preparation seems to offer advantages for the study of antibody-antigen complexes. Under the conditions of our experiments, the surfaces of the antigenic structures are saturated with antibody layers approximately 95 A in thickness, i.e., slightly less than half the accepted lengths of 7S antibody molecules. Our interpretation is that the antibody is attached by combining sites at the ends of the molecules to form loops along the surface of the antigenic structures. 相似文献
970.
Seiichi Ishizuka Sachio Ishimoto Anthony W. Norman 《Archives of biochemistry and biophysics》1982,217(1):264-272
Kidney homogenates from vitamin D3-supplemented chicks incubated with 25-hydroxyvitamin D3 [25(OH)D3] produce significant quantities of a new, unknown vitamin D metabolite. This metabolite was isolated in pure form from such incubation mixtures by using Sephadex LH-20 column chromatography followed by high-pressure liquid chromatography. This metabolite has been identified as 23,25,26-trihydroxyvitamin D3 [23,25,26(OH)3D3] by loss of radioactivity from 25-hydroxy[23,24-3H]vitamin D3 and 25-hydroxy-[26,27-methyl-3H]vitamin D3, ultraviolet absorption spectrophotometry, mass spectrometry, and periodate cleavage oxidation followed by mass spectrometry. This same metabolite was also isolated from the serum of rats given large doses of vitamin D3, and structurally characterized as 23,25,26-trihydroxyvitamin D3. As yet, the stereochemistry at the C-23 and C-25 positions of the natural product remains unknown. A comparison of responses to a single dose level (500 ng) of 23,25,26(OH)3D3 or 25(OH)D3 over 96 h in vitamin D-deficient rats indicated that the new metabolite had no capability to mediate bone calcium mobilization and that it was only weakly active in stimulating intestinal calcium transport. 相似文献