Biomedical subjects
Publications and source records attributed to E Hoshino.
We describe Eubacterium minutum sp. nov., which was isolated from human periodontal pockets. This new species was established on the basis of DNA-DNA hybridization data. The guanine-plus-cytosine content of its DNA is 38 to 40 mol%. The results of differential biochemical and enzymatic tests are described. The type strain of this species is strain M-6.
We compared the usefulness of lansoprazole and famotidine for treatment of reflux esophagitis. Fifty-four subjects with moderate to severe reflux esophagitis were divided into two groups. The lansoprazole group (LPZ) received lansoprazole 30 mg once daily for 4 weeks, followed by famotidine 20 mg twice daily for 4 weeks. The famotidine group (FAM) received the same medications in the reverse order. Patient evaluation was based on six factors found on endoscopy, and on patient reports of heartburn. Evaluations were done before the start of the study and at 4 and 8 weeks. The improvement rates in the grade of GI classification at 4 weeks were 57.7% in the LPZ group and 38.5% in the FAM group; there was no significant difference between the two groups. However, after drugs were switched, more patients who switched to famotidine experienced a deterioration of their condition than did those who switched to lansoprazole. The percentages of "useful" or "very useful" were not significantly different between the two groups, but the percentage of "very useful" in the LPZ group (46.2%) was significantly higher than that in the FAM group (14.3%) (p < 0.05). These results suggest that lansoprazole is the first drug of choice for treatment of moderate to severe reflux esophagitis.
Seventy-one patients with peptic ulcer (44 gastric ulcer and 27 duodenal ulcer patients) were enrolled in this study. After the ulcers had healed with 2 months of treatment with lansoprazole (LPZ) 30 mg/day, patients were divided into two groups: 43 in the half-dose q.d. group and 28 in the weekend full-dose group. Fourteen (23%) of the 43 patients in the half-dose q.d. group and 14 of the 28 (50%) in the weekend full-dose group failed to continue taking the drug for 6 months owing to poor compliance (p < 0.05). None of the 30 patients who continued to take the maintenance dose of LPZ experienced ulcer recurrence during the 6-month follow-up period, whereas 71% of the patients who stopped taking LPZ experienced recurrence (p < 0.01). In conclusion, maintenance therapy using the half-dose q.d. administration schedule was superior to weekend full-dose administration because of the better patient compliance and a lower ulcer recurrence rate.
Appendiceal cancer associated with pseudomyxoma peritonei is a low grade malignancy and its extraperitoneal metastasis is extremely rare. We report a case of gastric metastasis of this tumor in a 76-year-old man. Two metastatic gastric tumors, which appeared after a 1-year interval, were successfully resected endoscopically. The patient was well for more than 3 years after the onset of the disease. To our knowledge, gastric metastasis from appendiceal cancer with pseudomyxoma peritonei has not been previously reported.
Degradation of phenylalanine and leucine by resting cells of Eubacterium brachy ATCC 33089 were studied under strict anaerobic conditions. The effects of metronidazole and air on the metabolism were also studied. The organism principally produced phenylpropionate and isocaproate from phenylalanine and leucine, respectively. Other products were cinnamate and phenylacetate from phenylalanine, and alpha-ketoisocaproate and isovalerate from leucine. The organism also produced hydroxylated end-products, i.e. phenyllactate from phenylalanine and hydroxyisocaproate from leucine. When metronidazole was added to the reaction mixture, the production of phenylpropionate, cinnamate, phenylacetate, isocaproate, alpha-ketoisocaproate and isovalerate was inhibited, while that of hydroxylated products was not, suggesting that the organism has metronidazole-sensitive and -tolerant pathways of metabolism. A similar inhibitory effect was also found when the reaction was done aerobically, suggesting that the inhibitory mode of metronidazole is similar to that of oxygen.
The aim of this study was to determine whether microorganisms invade periapical cementum of human teeth from the adjacent periapical lesions. We therefore attempted to isolate microorganisms from periapical cementum through the adoption of standard anaerobic procedures for obligate anaerobes. Samples of cementum were taken from 10 amputated tooth roots at the time of apicoectomy. From two of these samples, bacteria were recovered after anaerobic incubation, but no bacteria were recovered after aerobic incubation of the same samples. Of a total of eight isolates from the cementum, seven were obligate anaerobes and one was aerotolerant. The obligate anaerobes isolated were assigned to the genera Prevotella, Peptostreptococcus, Eubacterium, and Fusobacterium. The aerotolerant anaerobe was Campylobacter. From this, we conclude that bacteria can successfully invade cementum via periapical periodontal tissue, and that such bacteria may play a significant role in chronic periapical pathosis.
The mode of hydrolysis of cotton cellulose by two highly purified exo- and endo-type cellulases from Irpex lacteus was investigated by differential scanning calorimetry, to measure changes in the size of the amorphous region in cotton fibers with the enzymatic reaction. The cellulases induced entirely different changes in the size of the amorphous region, particularly at earlier stages of reaction. Exo-type cellulase gradually reduced the amorphous region with release of cellobiose from the initial stage of hydrolysis, but began to increase the amorphous region at more advanced stages of hydrolysis. By contrast, endo-type cellulase caused no liberation of reducing sugar at the initial stage of hydrolysis but caused a sharp increase in the amorphous region, and it thereafter caused a rapid decrease of the amorphous region, accompanied with the production of various kinds of cellooligosaccharides. The rate of size reduction of the amorphous region caused by endo-type cellulase was much higher than that by exo-type cellulase. Convergence of the decrease in the size of amorphous region during hydrolysis by endo-type cellulase is followed by the increase in this region being influenced by further hydrolysis of remained crystalline region. Substantial changes in the morphology of cotton occurred with the two cellulases after the hydrolysis stages at which the size of the amorphous region was minimum.
The antigenic polysaccharide produced by Eubacterium saburreum, strain T19, contains unusual sugars, including D-glycero-D-galacto-heptose (Hep) and D-fucose (D-Fuc). A repeating unit of the polysaccharide is composed of a linear chain of D-glycero-D-galacto-heptopyranosyl tetrasaccharide as its backbone structure, i.e., -[-->6)-beta-Hep p-(1-->3)-beta-Hep p-(1-]2-->, and a D-fucofuranosyl disaccharide as a branched group, i.e., alpha-D-Fuc f-(1-->2)-alpha-D-Fuc f-(1-->, which is linked to O-4 of one (1--6)-linked D-glycero-D-galacto-heptopyranosyl residue. The polysaccharide also contains O-acetyl groups.
The mode of action of two highly purified cellulases of exo- and endo-types from Irpex lacteus was investigated by using pure cellulosic substrates with different crystallinities derived from cellulose I. Exo-type cellulase saccharified all celluloses more effectively than endo-type enzyme, and the saccharification activities of both cellulases similarly increased with decreasing crystallinity of cellulose. The DP-lowering activity of exo-type cellulase remained similar for celluloses with higher crystallinity, while this cellulase showed a degradation mode resembling that of the endo-type enzyme for the substrates with lower crystallinity. Compared with exo-type cellulase, endo-type cellulase remarkably decreased the DP of cellulose with higher crystallinity, while this activity was abated for cellulose with lower crystallinity. Thus, the effects of both cellulases became similar in the degradation of amorphous substrates such as H3PO4-treated cellulose. Endo-type cellulase produced several kinds of cellooligosaccharide from all kinds of cellulose used, while the product of the exo-type enzyme was only cellobiose from crystalline cellulose such as cotton and cotton linter even after a 12-h incubation period. The results indicate that each cellulase shows a typical mode of action (exo or endo) for crystalline cellulose, but that their characteristic modes of attack may change with decreasing crystallinity of cellulose.
The interaction of two highly purified cellulases, exo- and endo-type cellulases from Irpex lacteus, with pure cotton and amorphous cellulose was investigated by electron microscopy. The morphological observations indicated that exo- and endo-type cellulases are both strongly adsorbed on the internal microfibril of cotton fiber before enzymatic hydrolysis, and then initiate their action toward the internal cellulose microfibrils with retention of the original shape. The two cellulases, however, caused considerably different morphological changes in cotton cellulose, and each cellulase seems to degrade native cellulose with a distinct mode of action. In the hydrolysis of cotton with exo-type cellulase, deep transverse cracks were produced and they extended from the fiber surface to the lumen structure located inside the fibers. In contrast, it was found that there were no deep cracks on fibers treated with endo-type cellulase, but severe internal erosion and cavitation occurred along fibril or microfibril layers inside the fibers. Thus, the degradation of cotton by exo- and endo-type cellulases yielded quite different morphological patterns, while little difference was found for regenerated celluloses. The mode of enzymatic hydrolysis of cellulose shown by cellulases with different degrees of randomness (exo and endo types) appears to be markedly affected by the fine structure of cellulose fibers.
A new species, Eubacterium saphenum [corrected] sp. nov., established on the basis of the results of DNA-DNA hybridization, was proposed for strains isolated from human periodontal pockets. Differential characteristics are given.
Antigens of Eubacterium species including E. alactolyticum, E. brachy, E. nodatum, E. saburreum, E. timidum, E. yurii subsp. yurii and E. yurii subsp. margaretiae, which have been isolated frequently from periodontal pockets and associated with periodontal diseases, were extracted by ultrasonication from whole bacterial cells. Antigens were also prepared from E. aerofaciens, E. lentum and E. rectale, which have been found in intestinal tracts and infected abscesses in human oral cavities. The antigens of the oral Eubacterium species were compared with antigens from E. limosum, the type species of the genus Eubacterium, by using SDS-PAGE and Western immunoblot assays. SDS-PAGE gels stained with Coomassie brilliant blue indicated that no major peptide bands were common among the Eubacterium species examined. The protein profile patterns were distinctly different from each other. Western immunoblotting reactions with rabbit antisera showed that the Eubacterium species could be clearly distinguished serologically, and that the species-specific antigens were peptide components of ultrasonic extracts from the whole bacterial cells. The present study demonstrates that these Eubacterium species show great heterogeneity in their peptide components and immunological reactions, which may be useful for identification of the Eubacterium species from human oral specimens.
The purpose of this study was to clarify the antibacterial efficacy of mixed antibacterial drugs on bacteria of carious and endodontic lesions of human deciduous teeth in vitro. The antibacterial drugs used in this study were mixtures of ciprofloxacin, metronidazole, plus a third antibiotic: amoxicillin, cefaclor, cefroxadine, fosfomycin or rokitamycin. Samples taken from carious dentin (17 cases) and infected pulpal tissues (14 cases) were cultured on control plates and plates containing the mixed drugs. No bacteria were recovered in the presence of any combination of the mixture of the drugs (100 micrograms each/ml), and the bacterial growth occurred on control plates (10(1) to 10(7) colony-forming units), indicating that the mixed drugs inhibit the growth of bacteria in the samples. When carious and endodontic lesions on split surfaces of freshly extracted teeth were covered overnight with alpha-tricalcium phosphate cement containing a mixture of ciprofloxacin, metronidazole and cefaclor (1% each; 5 cases), no bacteria were recovered from the lesions. No bacteria were recovered from carious and endodontic lesions when these lesions were immersed in a solution of the mixture (200 micrograms each/ml; 5 cases). These findings indicate that carious and endodontic lesions can be sterilized by the mixed drugs in situ.
The adsorption mode of two highly purified cellulases, exo- and endo-type cellulases, from Irpex lacteus (Polyporus tulipiferae) was investigated by using pure cellulosic materials with different crystallinity as substrates. Adsorption of the two enzymes on the substrates was found to fit the Langmuir-type adsorption isotherm. Maximum amount of adsorbed enzyme obtained from the Langmuir plots showed an inverse correlation to the crystallinity of the substrate with both enzymes, and this value of endo-type cellulase was less dependent on the degree of crystallinity of substrates than that of exo-type cellulase, whose isotherms reached saturation in the range of low enzyme concentrations. The two enzymes showed relatively high affinities for all the substrates and their affinities increased with increasing crystallinity, but this tendency was less marked with endo-type cellulase than with exo-type one. In addition, large negative values of free energy change were observed on the adsorption of both enzymes, and the values became more negative with increasing crystallinity. Consequently, both cellulases showed high adsorption on crystalline cellulose and the adsorption process became smoother with increasing crystallinity. The adsorption of the two types of cellulases was endothermic with an increase in entropy, especially for amorphous cellulose, suggesting the occurrence of water release from the substrates during enzyme adsorption. In addition, the changes in thermodynamic parameters (delta H, delta S, and delta G) in adsorption of exo-type cellulase were larger than in that of endo-type enzyme.
Anaerobic procedures were adopted to demonstrate the early bacterial invasion of non-exposed dental pulps, and to isolate and identify the bacteria. Of 19 freshly extracted teeth which originally exhibited deep dentinal lesions, clinical examination and electric pulp testing showed that nine of them had no pulpal exposure. Thus the pulps of these teeth were covered by clinically sound dentine beneath the carious lesion. Bacteria were found to have invaded the pulps of six of these nine teeth. The predominant bacteria were obligate anaerobes belonging to the genera Eubacterium, Propionibacterium and Actinomyces. Other obligate anaerobes were Lactobacillus, Peptostreptococcus, Veillonella and Streptococcus. The bacterial composition resembled that of the deep layers of dentinal lesions described previously, suggesting that the bacteria isolated in this study had passed through some individual dentinal tubules, to invade the dental pulp.
An antigenic surface polysaccharide produced by Eubacterium saburreum strain T18, isolated from human dental plaque, was purified from formamide extract of whole cells. Methylation analysis, Smith degradation, optical rotation data and nuclear magnetic resonance spectra demonstrated that the purified antigen was a homopolysaccharide composed of D-glycero-D-galacto-heptose (Hep.) residues. The structure of the repeating unit in the polysaccharide was: -[----6)-[alpha-Hep.furanosyl-(1----4)]-beta-Hep.pyranosyl- (1----6)-[alpha-Hep.furanosyl-(1----2), alpha-Hep.furanosyl-(1----4)]-beta- Hep.pyranosyl-(1-)4----6)-beta-Hep.pyranosyl-(1----. No heptose residues were acetylated. Immunodiffusion reactions in agar gel suggested that the immunodeterminant of the antigenic polysaccharide was D-glycero-D-galacto-heptofuranosyl residues as branched nonreducing terminals.
This study was carried out to investigate the predominant anaerobic bacteria of periodontal pockets in patients with advanced periodontitis, who had no previous treatment other than supragingival scaling, no history of recent or chronic systemic illness, nor any intake of antibiotics within 6 weeks prior to bacteriological sampling. Care was taken not to ignore tiny-colony-forming anaerobes, by means of a stereoscope and an anaerobic glove box system. Out of 422 (100%) isolates, 380 (90%) were obligate anaerobes, suggesting that the environment in periodontal pockets was anaerobic and favors the growth of obligate anaerobes. Among the 380 obligate anaerobes isolated, strains belonging to Eubacterium (54%) were predominant, and many of them occurred in tiny colonies. The other obligate anaerobes isolated were assigned to Wolinella (9%), unidentified motile rods which resemble Wolinella (7%), Peptostreptococcus (6%), Fusobacterium (5%), Bacteroides (2%; including those reclassified to Prevotella and Porphyromonas) and Selenomonas (0.5%). Among the isolates, 67% were Gram-positive bacteria, including 59% of rods (mostly asaccharolytic Eubacterium), suggesting that these bacteria, particularly strains of the Eubacterium species, may play an important role in etiology of adult periodontitis.
The circadian rhythm of respiratory function was examined in normal males and asthmatic subjects. In both groups, maximal flow decreased in the early morning. The major factor contributing to the circadian change in V50 was lung elastic recoil pressure in normal males and upstream resistance in asthmatic subjects. In some asthmatic patients, beclomethasone inhaled in the afternoon and at night was more effective than in the morning in increasing both the circadian maximal and minimal peak expiratory flow rates. This indicates the importance of chronotherapy in asthma.