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Biomedical subjects

Y Aoi

Publications and source records attributed to Y Aoi.

At least 19 recordsLinked to original sources

Flavin reductase coupling with two monooxygenases involved in dibenzothiophene desulfurization: purification and characterization from a non-desulfurizing bacterium, Paenibacillus polymyxa A-1.

The dibenzothiophene (DBT) desulfurizing bacterium metabolizes DBT to form 2-hydroxybiphenyl without breaking the carbon skeleton. Of the DBT desulfurization enzymes, DszC and DszA catalyze monooxygenation reactions, both requiring flavin reductase. We searched for non-DBT-desulfurizing microorganisms producing a flavin reductase that couples more efficiently with DszC than that produced by the DBT desulfurizing bacterium Rhodococcus erythropolis D-1, and found Paenibacillus polymyxa A-1 to be a promising strain. The enzyme was purified to complete homogeneity. K(m) values for FMN and NADH were 2.1 microM and 0.57 mM, respectively. Flavin compounds were good substrates, some nitroaromatic compounds were also active, and regarding the electron donor, the activity for NADPH was about 1.5 times that for NADH. In the coupling assay with DszC, only FMN or riboflavin acted as the electron acceptor. The coupling reactions of P. polymyxa A-1 flavin reductase with DszC and DszA proceeded more efficiently (3.5- and 5-fold, respectively) than those of R. erythropolis D-1 flavin reductase when identical enzyme activities of each flavin reductase were added to the reaction mixture. The result of the coupling reaction suggested that, in the microbial DBT desulfurization, flavin reductase from the non-DBT-desulfurizing bacterium was superior to that from the DBT-desulfurizing bacterium.

Amino Acid Sequence↗

Real-time monitoring of ammonia-oxidizing activity in a nitrifying biofilm by amoA mRNA analysis.

Ammonia monooxygenase encoding mRNA (amoA mRNA) transcription in the wastewater treatment process was investigated using reverse transcription PCR (RT-PCR) as the model indicating specific function and activity in nitrifying processes. The dynamic response of amoA mRNA transcription and ammonia-oxidizing activity to the change of environmental conditions such as pH and concentration of ammonia was examined to determine the inductive factor and the inhibitor for amoA mRNA expression. Furthermore, we semiquantitatively investigated the response of amoA mRNA transcription to the pH fluctuation in a continuous fed nitrifying reactor. As a result, amoA mRNA oriented analysis enabled real-time assay of ammonia-oxidizing activity within 2 h as a response time. In contrast, rRNA and amoA encoding DNA were constantly detected at almost the same amount throughout the experiment. mRNA transcription was regulated by the many environmental conditions: ammonia seems to be one of the strong inducers for transcription of amoA mRNA, whereas low pH seems to be a strong inhibitor. These factors simultaneously affected the mRNA transcription and enzymatic activity leading to the complex phenomena of ammonia-oxidizing activity and amoA mRNA transcription in the continuous feeding reactors.

Ammonia↗

Isolation of Pseudomonas aeruginosa from Ushubetsu River water in Hokkaido, Japan.

To provide information on the ecology of Pseudomonas aeruginosa in nature, bacteriological surveillance was performed in the defined area in Hokkaido, Japan. P. aeruginosa was isolated from water samples of Ushubetsu River in the down stream from the urban area of Asahikawa. P. aeruginosa was isolated from fecal samples of pigs but not from samples of soil of a tomato field, sand of sandboxes in vest-pocket parks, fresh vegetables, or feces of wild deer. The present results indicate that P. aeruginosa strains isolated from the river water is originated from the environment of human activity and not from wild life or domestic animals.

Animals↗

Ligation errors in DNA computing.

DNA computing is a novel method of computing proposed by Adleman (1994), in which the data is encoded in the sequences of oligonucleotides. Massively parallel reactions between oligonucleotides are expected to make it possible to solve huge problems. In this study, reliability of the ligation process employed in the DNA computing is tested by estimating the error rate at which wrong oligonucleotides are ligated. Ligation of wrong oligonucleotides would result in a wrong answer in the DNA computing. The dependence of the error rate on the number of mismatches between oligonucleotides and on the combination of bases is investigated.

Animals↗

Oral flora of mongrel and beagle dogs with periodontal disease.

The plaque flora was studied in adult mongrel and beagle dogs with periodontal disease. Gingival plaque from maxillary premolars was removed and cultured on various growth media. The flora in all dogs was composed of mostly anaerobic gram negative rods. Bacteroides asaccharolyticus was found in the highest proportion of plaque samples from mongrel dogs, and decomposed hydrogen peroxide suggesting catalase activity. Fusobacterium nucleatum was found in higher proportion in the plaque of beagle dogs as compared to B. asaccharolyticus. With the increasing numbers of obligative anaerobic gram negative organisms such as B. asaccharolyticus, the proportions of Streptococcus, Enterococcus and Staphylococcus decreased in the dogs with periodontal disease. The salivary flora was different from the plaque flora of the dogs with periodontal disease. It was constant regardless with the disease. The salivary flora of beagle dogs with the healthy gingiva was different from that of mongrel dogs. Enterococcus, Lactobacillus, Eubacterium and black-pigmented Bacteroides (BPB, mainly B. asaccharolyticus) were higher proportion in the flora of beagle dogs as compared to mongrel dogs, while Fusobacterium, Enterobacteriaceae, yeast and molds were lower in the flora. The results reveal that B. asaccharolyticus and F. nucleatum are common pathogens and uniquely contribute to the development of gingival inflammation in dog.

Animals↗

Epidemiological study on periodontal diseases and some other dental disorders in dogs.

The prevalence of dental disorders in dogs was studied by applying index systems for human with some modifications. A total of 251 mongrel dogs including 143 stray dogs kept in the Animal Protection Offices in Tokyo and Hokkaido and 108 pet dogs visiting veterinary clinicians in Chiba Prefecture and Hokkaido were used. Periodontitis was prevalent among these dogs regardless of their sources and its incidence was increased with age. The lesion was more severe and more frequent in the premolar and molar regions than in the maxillary and mandibular incisor regions. Missing of teeth was observed at a high and increasing incidence with age. The tooth most commonly lost was the first premolar, followed by the other premolars and molars, where severe periodontitis was frequently found. Calculus was seen on many teeth, and aging agravated its prevalence and severity. Dental caries was observed in stray dogs, but neither to a serious degree nor at a significant level. These findings emphasize the necessity of dental hygiene, proper dental care and continuous periodical survey for dogs.

Age Factors↗

Chemical analysis of glycosaminoglycans inhibiting DNA synthesis.

Sulfated glycosaminoglycans having inhibitory activity in cellular and subcellular systems were found in some tumor tissues from humans. These glycosaminoglycans inhibited more efficiently DNA synthesis of virus transformed cells (SV40-WIRL-3 cells) than their parent normal cells (WIRL-3 cells). Sulfated glycosaminoglycans found in normal human and non-tumor tissues did not have as high an inhibitory activity on DNA synthesis by cells used in this investigation as those from some human tumor tissues. The former did not inhibit as effectively DNA synthesis by virus transformed cells, as DNA synthesis by their normal parent cells. The monosaccharide composition of these sulfated glycosaminoglycans showed N-acetyl glucosamine (Glu-NAc) as a main monosaccharide, and xylose (Xyl), glucose (Glu), galactose (Gal), hyaluronic acid (Hu-A) as minor monosaccharides. N-acetyl galactosamine was not detected.

Adult↗

Protection against hemorrhagic pneumonia of mink by Pseudomonas aeruginosa multicomponent vaccine.

An attempt to prevent epidemics of hemorrhagic pneumonia in mink due to Pseudomonas aeruginosa was made in the course of epidemics with injection of the multicomponent vaccine consisting of common protective antigen (OEP) of P. aeruginosa mixed with toxoids of protease and elastase of the bacillus. Enzootics of hemorrhagic pneumonia, due to P. aeruginosa serotype 8, broke out from August to October 1977 in a total of 13 sheds of 3 farms (A, B and C) which were located in the northeast area of Hokkaido. These farms were raising 7,452, 2,553 and 10,639 mink respectively. The mortality rate of the mink on farms A, B and C were 11.8%, 13.0% and 1.0% respectively. The vaccination was performed on the 3 farms 5, 8 and 21 days after the onset of the disease. Inoculation of each mink with 200 micrograms or 100 micrograms of each of the three components of the multicomponent vaccine was effective in most of the male and female groups of mink. The period required for revealing the effect of the vaccination was very short, in some cases only a few days. Administration of the vaccine 21 days after the onset of the enzootic was also effective.

Animals↗

Surface glycopeptide change triggered by contact between normal cells from rat liver and their simian virus 40-transformed cells from the same virus.

Surface glycopeptide changes triggered by mixed culture of normal cells from rat liver cell line WIRL-3 and simian virus 40-transformed cells from the same line were studied. In addition, biologic responses of the normal cells and the transformed cells to three kinds of surface glycopeptides derived from the normal cells, the transformed cells, and these cells in mixed culture were investigated. Surface glycopeptide derived from the mixed culture showed a quantitative increase and a qualitative change in the glycopeptide structure, not detected in the surface glycopeptide of normal cells and transformed cells cultured separately. The surface glycopeptide changes observed in the mixed culture were mainly ascribable to normal cells responding to transformed cells, and a surface glycopeptide change in normal cells induced a qualitative surface glycopeptide change in transformed cells. Differences in biologic response of normal cells and transformed cells to the above three surface glycopeptides were investigated with the use of inhibition of [3H]thymidine uptake of the cells by the three different surface glycopeptide preparations. The inhibition rates of uptake in normal cells were 30% by all three surface glycopeptides (200 microgram/ml culture medium); inhibition rates in transformed cells were 65% by mixed-culture surface glycopeptide, 50% by normal cell-surface glycopeptide, and 35% by transformed cell-surface glycopeptide.

Cell Communication↗

Biosynthesis of glycoprotein-glycosyl transferases during the cell cycle.

The levels of glycoprotein-glycosyl transferases increased at the late G1-early S phase during the cell cycle both in the plasma membrane and in the microsomal fraction. The levels of the enzyme showed a "peak pattern" during the cell cycle. Elevation of the enzyme activity at earlier stages was not influenced by cytosine arabinoside but that at late G1 was completely inhibited.

Animals↗

Alterations in surface glycoproteins and level of sialyltransferase activity of human embryo kidney cells infected with oncogenic adenovirus type 12.

Human embryo kidney (HEK) cells infected with adenovirus type 12 and its cytocidal mutants (Cyt) synthesized different glycoproteins respectively on the cell surface at the early stage of infection. Furthermore, a less and a highly tumorigenic Cyt mutants synthesized different glycoproteins. The level of sialyltransferase activity in the microsomes of HEK cells was increased when cells were infected with adenovirus type 12. It was confirmed that there was a positive correlation between the sialyltransferase activity in the microsomes and tumorigenicity.

Adenoviridae↗