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Biomedical subjects

T Fujiwara

Publications and source records attributed to T Fujiwara.

At least 919 records · Page 51Linked to original sources

Chemical, immunobiological and antigenic characterizations of lipopolysaccharides from Bacteroides gingivalis strains.

Lipopolysaccharides (LPS) were extracted from whole cells of seven strains of Bacteroides gingivalis--381, ATCC 33277, BH18/10, OMZ314, OMZ406, 6/26 and HW24D-1--by the phenol/water procedure, and purified by treatment with nuclease and by repeated ultracentrifugation. These LPS were composed of hexoses, hexosamines, fatty acids, phosphorus and phosphorylated 2-keto-3-deoxyoctonate (KDO). The major components of the lipid portion of these LPS were hexadecanoic, 3-hydroxyhexadecanoic, branched 3-hydroxypentadecanoic and branched 3-hydroxyheptadecanoic acids. All the LPS preparations induced marked mitogenic and in vitro polyclonal B cell activation responses in spleen cells from both C3H/HeN and C3H/HeJ mice, exhibited no definitive preparatory activity in the local Shwartzman reaction in rabbits, but were active in the chromogenic Limulus amoebocyte lysate test. A monoclonal antibody (mAb) raised against the LPS from B. gingivalis strain 6/26 reacted with LPS from all other B. gingivalis strains tested. Other mAbs raised against LPS from B. gingivalis strains 381 and 6/26 reacted with the LPS from strains 381, ATCC 33277, BH18/10 and 6/26 (these strains were termed LPS serogroup I), as revealed by ELISA and immunodiffusion. The LPS from these strains except for 6/26 showed almost identical patterns in SDS-PAGE stained with ammoniacal silver. A mAb raised against the LPS from B. gingivalis HW24D-1 reacted with the LPS from strains OMZ314, HW24D-1 and OMZ409 (LPS serogroup II). These LPS, except OMZ409, exhibited very similar profiles in SDS-PAGE. These results indicate that there are at least two different antigenic groups present among LPS from B. gingivalis strains, as well as a common, species-specific antigen.

Animals↗

The arcB gene of Escherichia coli encodes a sensor-regulator protein for anaerobic repression of the arc modulon.

The arcA (dye) and arcB genes of Escherichia coli are responsible for anaerobic repression of target operons and regulons of aerobic function (the arc modulon). The amino acid sequence of ArcA (Dye) indicated that it is the regulator protein of a two-component control system. Here we show that ArcB is a membrane sensor protein on the basis of its deduced amino acid sequence (778 residues), hydropathicity profile, and cellular distribution. On the carboxyl end of the ArcB sequence there is an additional domain showing homology with conserved regions of regulator proteins. Deletion into this domain destroyed ArcB function. ArcB conserved a histidine residue for autophosphorylation of the sensor proteins, and aspartic residues important for the regulator proteins.

Amino Acid Sequence↗

Antimicrobial activities of thiolactomycin against gram-negative anaerobes associated with periodontal disease. f1.

Thiolactomycin (TLM), (4R)-(2E,5E)-2,4,6-trimethyl-3-hydroxy-2,5, 7-octatriene-4-thiolide, purified from a culture filtrate of a strain of the Nocardia species, was examined for antimicrobial activities against more than 100 strains of oral and periodontally associated bacteria. Nine other commonly used antibiotics were also included for the test. We found that TLM exhibited strong and selective antimicrobial activities against Bacteroides gingivalis and other oral black-pigmented Bacteroides species that may be etiologically associated with adult periodontitis. TLM also inhibited the growth of Actinobacillus actinomycetemcomitans, but did not affect the growth of oral streptococcal species and Eubacterium species. Strains of Eikenella corrodens were moderately susceptible to TLM, while Actinomyces viscosus strains were only slightly susceptible to it. Other antibiotics used for comparison showed a broad spectrum of antimicrobial activities in general. In conclusion, TLM exhibited highly selective antimicrobial activities to black-pigmented Bacteroides species and A. actinomycetemcomitans, both of which are implicated in the pathogenesis of human periodontal disease.

Actinobacillus↗

Interaction of structural isomers of sucrose in the reaction between sucrose and glucosyltransferases from mutans streptococci.

Structural isomers of sucrose, i.e. disaccharides composed of glucose and fructose molecules with different glucosidic linkages, were examined for their effect on the reaction between sucrose and various glucosyltransferases (GTases) from Streptococcus mutans MT8148 and Streptococcus sobrinus 6715. Trehalulose (alpha 1-1), turanose (alpha 1-3), maltulose (alpha 1-4), and palatinose (alpha 1-6) were used as the sucrose analogues. Mutans streptococci were found not to utilize these sucrose analogues. Analysis of enzymatic products of GTase and sucrose with thin layer chromatography clearly revealed that glucan synthesis from [14C]sucrose by the various purified GTase preparations from S. mutans and S. sobrinus was inhibited in the presence of these sucrose analogues except turanose, resulting in the release of increased amounts of [14C]fructose and [14C]oligosaccharides. It was also found that the fructose residues in the oligosaccharides were derived from those of sucrose analogues but not sucrose itself. The Lineweaver-Burk plots of the substrate saturation kinetics of GTase vs sucrose indicated increased Km and Vmax in the presence of sucrose analogue, as compared with sucrose alone. Finally, these sucrose analogues except turanose inhibited sucrose dependent cellular adherence of S. sobrinus 6715 to a glass surface, while they scarcely inhibited the adherence of S. mutans MT8148. Among the analogues, maltulose appeared the most effective inhibitor against GTases in general.

Chromatography, Thin Layer↗

A case of epidural granulocytic sarcoma preceding acute leukemia.

A 20-year-old male developed both coccygeal and leg pain and followed by rectocystic disturbance. Disc herniation between L5 and S was suspected and laminectomy was performed. At surgery, an easily curretable tumor occupied the epidural space from L5 to the end of the sacrum. In part, the tumor spread out of the vertebral canal and invaded the surrounding muscle tissue. This muscle tissue and part of the lamina were checked histologically. Initial blood analysis revealed 5% blast-like cells, but failed to confirm them as leukemic cells. Histologically, the tumor cells had round or oval nuclei with large nucleoli and scanty cytoplasm without granulocytic differentiation. Malignant lymphoma or Ewing's sarcoma was initially suspected, but the definite diagnosis was uncertain. Immunohistochemical staining with the PAP method and enzyme histochemistry revealed that the tumor cells were positive for lysozyme and naphthol ASD chloracetate esterase. Thus, granulocytic sarcoma was finally diagnosed. Electron microscopic findings supported this diagnosis. Subsequent karyotyping of bone marrow cells revealed 8; 21 translocation, thus the final diagnosis of this patient was myelodysplastic syndrome, refractory anemia with excess blast cells in transformation or acute myelogenous leukemia, M2, by the FAB classification.

Acute Disease↗

Novel gelatin particle agglutination test for serodiagnosis of leprosy in the field.

We developed a novel gelatin particle agglutination test (MLPA) for the serodiagnosis of leprosy; this test is especially useful for clinical practice and epidemiological surveys of leprosy in countries in which the disease is endemic. The antigen used in the test is the chemically synthesized trisaccharide moiety of Mycobacterium leprae-specific phenolic glycolipid I. MLPA is a simple and easy technique having sensitivity and specificity comparable to those of the conventional indirect enzyme-linked immunosorbent assay. The new technique was found to be useful for monitoring of chemotherapy and predictive diagnosis of high-risk individuals in contact with persons with leprosy and may be useful for the prediction of relapse. We are now preparing to supply a quality-controlled ready-to-use MLPA kit for leprosy control in countries in which leprosy is endemic.

Agglutination Tests↗

Plaque formation of dietary isomaltulose in humans.

The plaque formation of isomaltulose, a sucrose isomer, was examined in 15 human volunteers with both diet and oral hygiene under supervision. The subjects were requested to refrain from all oral hygiene procedures for 3 days and were provided between-meal snacks containing 157 g of 4 test sugars (100% isomaltulose, 70% isomaltulose + 30% sucrose, 50% isomaltulose + 50% sucrose, and 100% sucrose). The study was repeated 4 times over 4 weeks. The isomaltulose diet resulted in the lowest plaque index, while sucrose induced a significantly greater deposition. In the absence of sucrose-containing snacks, mutans streptococci in saliva decreased below the baseline level. These results suggest that isomaltulose may be a suitable substitute for sucrose in between meal snacks.

Adult↗

Effect of experimental hyposalivation on the induction of dental caries in rats infected with Streptococcus mutans.

The effects of hyposalivation on the induction of dental caries were examined in specific pathogen-free Sprague-Dawley rats infected with Streptococcus mutans MT8148R and fed the caries-inducing diet 2000. Hyposalivation was induced by either irradiation or surgical removal of salivary glands in 30-day-old rats. After either procedure, severe dental caries was induced in the rats when examined at 84 days of age, and a statistically significant correlation was found between caries scores and flow rates of saliva. However, the recovery of strain MT8148R from hyposalivated rats was not always high, and it is possible that microorganisms other than the inoculated S. mutans strain MT8148R may have played a role in promoting dental caries.

Animals↗

Classification system for cataracts. Application by the Japanese Cooperative Cataract Epidemiology Study Group.

A cataract classification and grading system for the main purpose of cataract epidemiological studies is proposed by the Japanese Cooperative Cataract Epidemiology Study Group. Cataractous types are classified principally into cortical, nuclear and subcapsular opacities. Stages of cataract advancement are classified into grade I (early stage), grade II (moderate stage) and grade III (advanced stage). Regarding cortical opacities an opacified area obtained from red-reflex images is utilized for grading judgment. Nuclear opacity grading is based on the intensity of scattering at the nucleus. Grading of subcapsular opacities is judged from their extension in the three different pupillary areas. To help grading judgments, standard pictures of cortical and nuclear cataracts are provided. Gradings of lens coloration are divided into pale yellow (grade I), yellow (II), brownish yellow (III) and brown including reddish and black brown (IV). A recording chart is also provided.

Adult↗

Diabetes as risk factor of cataract: differentiation by retroillumination photography and image analysis.

We performed an epidemiological study to identify risk factors of cataract based on 2,243 subjects who underwent a general health checkup. Each subject underwent a 75-gram oral glucose tolerance test and blood chemistry analysis. The prevalence rate of cataract was higher in diabetic subjects in their 50s than in nondiabetics in the same age range (p less than 0.01). The incidence of vesicular and radial lens opacity was higher in diabetics than in nondiabetics (p less than 0.05).

Adult↗

Hematoma of the interatrial septum associated with complete atrioventricular block.

A 70-year-old man who had been treated for tongue cancer began to suffer from repeated syncopal attacks. His electrocardiogram indicated complete atrioventricular block. A permanent pacemaker was implanted. He was well for about 2 years after which time he developed dyspnea, suffered from general fatigue, and then suddenly died. A postmortem examination revealed a hematoma approximately 4 x 4. cm in size in the interatrial septum, connected to a noncoronary sinus and rupture of the noncoronary leaflet, which compressed the tricuspid and aortic valves. This hematoma might have been related to the conduction disturbance caused by destructive compression on the conduction system, and his death by the rupture of the noncoronary leaflet.

Aged↗

Attempts to quantity Clostridium botulinum type A toxin and antitoxin in serum of two cases of infant botulism in Japan.

Serum samples taken from two infant botulism cases during hospitalization were titrated for botulinum toxin by both the intraperitoneal (ip) injection method and the score method in mice. By the ip method, in which death is the only parameter, such low levels of toxin as lower than 4 ip LD50/ml may not be titrated even though the surviving mice show abdominal palsy. By the score method based on the degree of abdominal palsy, such low levels of toxin as 1.1 and 0.8 ip LD50/ml were detected in specimens of one of the patient's serum. No antitoxin was demonstrated in either case of infant botulism by applying the score method. It is not known whether spontaneous recovery from infant botulism is due to the antitoxin production.

Botulinum Antitoxin↗

Synthesis of RNA having cap structure.

RNA consisting 43 nucleotides bearing cap structure was synthesized (Figure). In the first place, 9 mer of a leader sequence with the cap structure (F-1) was synthesized by the phosphotriester method and followed by the capping reaction. Next, 32 mer of a cistron was divided into two fragments and each was synthesized by the phosphoramidite method. The 3'-end nucleotide of the RNA, a modified guanosine 5'-phosphate, was introduced to F-3 by use of P1-2',3'-O-methoxymethylene guanosine-5'-yl P2-adenosine-5'-yl diphosphate (A5' ppGmM) with T4 RNA ligase. The chemically synthesized RNA fragments were ligated with T4 RNA ligase to afford the desired RNA.

Amino Acid Sequence↗

DNA strand scission by neocarzinostatin: molecular recognition process responsible for site-specificity.

A series of hexanucleotides possessing A-T, G-C, inosine (I)-C and 2-aminoadenine (ANH2)-T base pairs at 5'-side of the target thymine were prepared and their selectivity for C-5' and C4' oxidation in the NCS-mediated degradation was investigated. Quantitative product analysis indicated that preferential C5' oxidation of deoxyribose moiety of the target T occurs at -5'-AT- and 5'-IT- sites, whereas C5' and C4' oxidation occurs competitively at T of -5'-GT- and -5'-ANH2T- sites. Based on the experimental results, an intercalation model that permits competitive hydrogen abstraction from C5' and C4' of deoxyribose moiety has been proposed.

Base Sequence↗