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S Hamada

Publications and source records attributed to S Hamada.

At least 19 recordsLinked to original sources

Molecular characterization and expression of the cell-associated glucosyltransferase gene from Streptococcus mutans.

A gene encoding cell-associated glucosyltransferase (CA-GTase) was cloned from Streptococcus mutans MT8148 into Escherichia coli DH5 alpha by using a low-copy-number plasmid, pMW119. After screening of a gene library with the oligonucleotide probe designed on the basis of a partial amino acid sequence of CA-GTase, a recombinant plasmid, pSK6, that had a 5.6 kb insert carrying the CA-GTase gene was selected. The gene product (recombinant CA-GTase) of pSK6 was expressed by using a lac promoter in pMW119. Western blotting revealed that rCA-GTase reacted with antibody to CA-GTase. rCA-GTase was found to synthesize water-insoluble glucans. Southern blotting indicated that the MT8148 chromosome contained another gene which was homologous to pSK6. A plasmid harboring this gene (pSK16) was also isolated from the gene library, the gene product of pSK16 exhibited GTase activity but ten times lower than that of pSK6.

Base Sequence

[Detectability of diagonal branch disease by 201TlCl exercised myocardial scintigraphy].

The detectability of diagonal branch disease in 10 patients (five with angina pectoris, five with myocardial infarction) with isolated diagonal branch lesions (more than 75% luminal stenosis in coronary angiography) was reviewed. In exercised 201TlCl myocardial scintigraphy, chest pain occurred in four of 10 patients, electrocardiographic change indicating myocardial ischemia was seen in four, and diagonal branch lesion was detected in only four patients by planar images. In contrast, diagonal branch lesions were detected in 10 of 10 patients by SPECT (single photon emission computed tomography). In planar images, perfusion defects appeared high in the anterolateral, posterolateral, and anterior walls of the left ventricle. In SPECT images they appeared high in the anterior to anterolateral wall. The extent of diagonal branch lesions could be quantitatively evaluated by coronary territory maps developed from unfolded maps of exercised SPECT. The mean ratio of the extent of diagonal branch lesion to left anterior descending branch territory was 24.7%, and the extent of myocardial infarction was significantly larger than that of angina pectoris (p < 0.05). In conclusion, SPECT is useful for detecting diagonal branch lesions and can quantitatively show the extent of these lesions by coronary territory map.

Aged

[Growth rate of abdominal aortic aneurysms as measured by Computerized tomography].

To evaluate the growth rate of abdominal aortic aneurysms, sequential computed tomography (CT) examinations were performed on 125 nonoperated patients (M/F 101/24, mean age 67 +/- 8 years, 129 lesions) at intervals of at least 6 months. Growth rates of aneurysms were obtained by subtracting the first from the last diameter and adjusting for examination interval. Mean growth rate was 0.28 +/- 0.26 cm/year. Aneurysms with an initial diameter exceeding 4 or 5 cm showed significantly faster growth than smaller aneurysms. There was no significant correlation between growth rate and atherosclerogenic factors, but systolic blood pressure was significantly higher in patients whose aneurysms ruptured. Our study showed quantitatively that aneurysms with a large diameter have a rapid growth rate, the growth rate of infrarenal aneurysms is significantly faster than that of suprarenal aneurysms, and blood pressure control is important to prevent rupture.

Adult

Induction of mucosal and serum immune responses to a specific antigen of periodontal bacteria.

The dynamics of the host immune response to periodontal bacteria not only may be informative from the standpoint of specific mucosal protection to these pathogens, but also may reveal the capacity of the mucosal immune response to provide protection of the host. To this end, we have examined the immune response to chromatographically purified fimbriae of P. gingivalis administered orally or systemically with liposomes and adjuvant in BALB/c mice, high responders to this antigen. Oral administration of P. gingivalis fimbriae clearly enhanced the fimbriae-specific salivary IgA response. ELISPOT analysis revealed that significant numbers of fimbriae-specific IgA SFC were seen in lamina propria and mesenteric lymph nodes but not in Peyer's patches of mice immunized orally. In contrast, antigen-specific IgM and IgG SFC were seen mainly in the circulating blood mononuclear cells. On the other hand, subcutaneous injection of fimbriae with GM-53 also raised the fimbriae-specific IgG followed by IgM and IgA responses in serum, and both IgA and IgG responses in saliva. Oral immunization was less effective than subcutaneous injection in terms of the serum antibody response. However, the salivary antibody level of mice injected subcutaneously was similar to that of mice immunized orally. In the subcutaneously immunized mice, fimbriae-specific SFC were detected in the spleen, blood, and brachial lymph nodes by ELISPOT assay. Fimbriae-specific IgM SFC appeared earlier and antigen-specific IgG SFC were seen later. These results show that the combined use of fimbriae together with the adjuvant results in sharply increased IgA responses in saliva and IgG responses in serum. In summary, it is clear that the nature of the host's antibody response in serum and mucosal secretions is distinct, and depends on the route of antigen administration, the use of adjuvant and/or liposomes, and the temporal phase of the humoral immune response following various immunization regimes.

Animals

Dependence of reaction rate of 5,5'-dithiobis-(2-nitrobenzoic acid) to free sulfhydryl groups of bovine serum albumin and ovalbumin on the protein conformations.

The effect of protein conformations on the reaction rate of Ellman's reagent, 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) with sulfhydryl (SH) groups of proteins was examined. The stopped-flow method was applied to follow the reaction of DTNB with SH group of two proteins, bovine serum albumin (BSA) and ovalbumin (OVA), at various concentrations of guanidine hydrochloride and urea. The rates for both the proteins were faster in guanidine than in urea. The rate sharply depended on the protein conformations, which were monitored by changes of helix contents on the basis of the circular dichroism measurements. The reaction rate of DTNB with SH groups of BSA was maximal around 2 M guanidine and 5 M urea. On the other hand, the reaction rate of DTNB with OVA was maximal at 3.5 M guanidine, while it gradually increased with an increase in the urea concentration. The amount of reactive SH group participating in the reaction with DTNB was also estimated by the absorbance change at 412 nm. The magnitudes of absorbance change for the reaction with free SH groups of OVA at low concentrations of the denaturants were appreciably smaller than those for BSA with one free SH group. Most of the four SH groups of OVA might react with DTNB above 5 M guanidine, although only a part of them did even at 9 M urea.

Dithionitrobenzoic Acid

Latex agglutination test for detection of mutans streptococci in relation to dental caries in children.

A simple and rapid system based on a latex agglutination (LA) reaction was devised for the detection of mutans streptococci in dental plaque. Latex particles were sensitized with antibodies against whole cells of Streptococcus mutans strains MT8148 (serotype c), MT703R (e) and OMZ175 (f) and Strep. sobrinus strains B13 (d) and 6715 (g). These sensitized particles were agglutinated within a few minutes after addition of 1.0-10 ng serotype-specific antigen from the homologous organisms or the nitrous acid extract of whole cells at 10(5)-10(6) c.f.u. The LA test specifically differentiated not only mutans streptococci from the other oral streptococci but also Strep. sobrinus from Strep. mutans. The LA test was also applicable to extracts of plaque from 206 human subjects who harboured mutans streptococci. In clinical trials, the outcome of the LA test correlated significantly with the number of mutans streptococci found in plaque (p less than 0.0001), which was quantified by the selective cultivation of mutans streptococci. Furthermore, the LA test discriminated between Strep. mutans and Strep. sobrinus from human dental plaque. The sensitivity and the specificity of the LA test for detection of mutans streptococci were 78.9 and 100%. The degree of reactivity in the LA test correlated significantly with the number of decayed tooth surfaces (p less than 0.0001) and decayed and filled tooth surfaces (p less than 0.0001). These results suggest that the LA test could be useful clinically for the detection of mutans streptococci in dental plaque as well as serving as a caries-activity test.

Antibodies, Bacterial

Osteochondritis dissecans of the humeral head.

A rare case of osteochondritis dissecans (OCD) occurring in the humeral head is presented. As there are only six available records in the literature concerning OCD in the humeral head, clinical features, treatment, and etiology of OCD involvement of this anatomical part remains unclear. These points are discussed on the basis of the previous cases and ours. Osteochondritis dissecans of the humeral head commonly affects middle-aged and young males rather than females. The prevalent location of involvement is the anterosuperior aspect of the humeral head. We propose that repetitive microtrauma is one of the important causes of OCD in the humeral head. Moreover, we suggest that the most authentic treatment for OCD with a partial detached osteochondral fragment is removal of this fragment and drilling.

Arthroscopy

Occurrence of antigen-specific B cells following oral or parenteral immunization with Porphyromonas gingivalis fimbriae.

The induction and distribution of antigen-specific antibody-secreting cells in various tissues were assessed in BALB/c mice immunized with the purified fimbrial protein of the Porphyromonas gingivalis strain 381. Groups of mice were immunized by gastric intubation of liposomes containing fimbriae and GM-53 on days 0, 1, 27, and 28. Additional groups of mice were immunized with P. gingivalis fimbriae and adjuvant GM-53 in Freund's incomplete adjuvant by subcutaneous injection on days 0 and 28. In the latter group of mice, levels of serum IgM anti-fimbria antibodies were first detected on day 7, while high levels of serum IgG anti-fimbria antibodies were seen after secondary immunization. Fimbria-specific spot-forming cells (SFC) were detected in the spleen, circulating blood mononuclear cells (CBMC), and brachial lymph nodes of immunized mice by ELISPOT. Fimbria-specific IgM SFC appeared by day 5 and antigen-specific IgG SFC were seen later in subcutaneously immunized mice. Mice immunized orally exhibited serum anti-fimbria IgG and IgA antibodies after boosting. Although numerical analysis revealed that the numbers of fimbria-specific SFC were generally lower than in subcutaneously immunized mice, significant numbers of antigen-specific IgA SFC were seen in lamina propria and mesenteric lymph nodes of orally immunized mice. In contrast, antigen-specific IgM and IgG SFC were observed mainly in CBMC. The route of immunization with fimbriae and GM-53 also influenced the total numbers of immunoglobulin-secreting cells. Thus, subcutaneous immunization enhanced the total number of IgM and IgG SFC, including fimbria-specific antibody-secreting cells in CBMC and the spleen. (ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral

Quantitative assessment of myocardial enhancement with iodinated contrast medium in patients with ischemic heart disease by using ultrafast x-ray computed tomography.

BACKGROUND AND RATIONALE: The authors report the quantitation of myocardial enhancement using iodinated contrast medium in patients with ischemic heart disease. Twenty-eight patients with chronic ischemic heart disease and 11 controls were examined by ultrafast computed tomography (CT) using 100-msecond scans. METHODS: The authors analyzed M/L (ratio of post-contrast incremental increases in the left ventricular myocardial and luminal CT number) in early and late phases after contrast injection. RESULTS: In controls, mean values of early and late M/L were 30% and 51%, respectively. In infarcted or severely ischemic segments, early M/Ls (19%, 16%) were significantly small (P less than .001), whereas late M/Ls (90%, 63%) were higher (P less than .001, .01) than controls. Segments with infarction or severe ischemia were differentiated from mild or nonsignificant ischemia by using this parameter (sensitivity, 99%; specificity, 88%). CONCLUSIONS: M/L is useful for detection of the ischemic myocardium.

Contrast Media

Purification and properties of extracellular mutacin, a bacteriocin from Streptococcus sobrinus.

Mutacin MT6223, a cell-free bacteriocin produced by Streptococcus sobrinus MT6223, was purified by ammonium sulphate precipitation, chromatofocusing with PBE 94 and column chromatography on SP Sephadex C-25. The specific activity of the purified mutacin was increased 1950-fold with a recovery of 9.7%. The molecular mass of the purified mutacin preparation was estimated to be 6.5 kDa. The mutacin activity was stable from pH 2-7, and was resistant to treatment at 100 degrees C for 20 min. It was inactivated by papain or ficin digestion, and was partially inhibited by alpha-chymotrypsin. The mutacin was found to be active against strains of serotypes c, e and f of Streptococcus mutans and the addition of purified mutacin MT6223 to growing cells of S. mutans MT8148 resulted in a rapid inhibition of incorporation of [3H]thymidine, [3H]uracil or L-[3H]glutamic acid into DNA, RNA or protein, respectively. Specific pathogen-free Fischer rats fed diet 2000 and infected with S. mutans MT8148R showed significantly fewer caries and lower plaque scores when mutacin was administered through drinking water. The present study demonstrates that mutacin MT6223 inhibited the growth of mutans streptococci. Thus, mutacin MT6223 may be a candidate for use in dental caries prevention.

Animals

Changes with time in the oral microflora and dental caries induction in hyposalivated rats fed on sucrose diet.

The effects of hyposalivation on the induction of dental caries and the change in oral microflora were examined at weekly intervals in Sprague-Dawley rats fed on diet 2000 containing 56% sucrose. In hyposalivated rats, significant dental caries was induced within one week and its severity increased with the experimental period. Bacteriological examinations demonstrated that the number of total cultivable microorganisms, lactobacilli and Staphylococcus aureus increased significantly shortly after surgical induction of hyposalivation, while the number of streptococci and yeasts did not increase significantly until the 7th week, beyond which time remarkable gross caries developed. A positive correlation was found between the caries score and the recovery of lactobacilli from mandibles of hyposalivated rats, while there was no statistically significant correlation between the caries score and the recoveries of S. aureus. On the other hand, dental caries was not induced in control rats fed on sucrose diet with no surgically-induced hyposalivation. It was also found that the number of lactobacilli increased significantly shortly after diet 2000 was given to control rats, but S. aureus was rarely recovered from the mandibles of control rats throughout the experiments. The roles of lactobacilli and S. aureus in the induction of dental caries under the hyposalivated condition were discussed and it was suggested that lactobacilli may play some significant role in the induction of dental caries in hyposalivated rats.

Animals

Immunochemical and biological characterization of outer membrane proteins of Porphyromonas endodontalis.

Outer membrane proteins (OMP) of Porphyromonas endodontalis HG 370 (ATCC 35406) were prepared from the cell envelope fraction of the organisms. The cell envelope that had been obtained by sonication of the whole cells was extracted in 2% lithium dodecyl sulfate and then successively chromatographed with Sephacryl S-200 HR and DEAE-Sepharose Fast Flow. Two OMP fractions, OMP-I and OMP-II, were obtained, and their immunochemical properties and induction of specific antibodies were examined. The OMP-I preparation consisted of a major protein with an apparent molecular mass of 31 kDa and other moderate to minor proteins of 40.3, 51.4, 67, and 71.6 kDa, while the OMP-II preparation contained 14-, 15.5-, 27-, and 44-kDa proteins as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis. OMP-I was found to form hydrophilic diffusion pores by incorporation into artificial liposomes composed of egg yolk phosphatidylcholine and dicetylphosphate, indicating that OMP-I exhibited significant porin activity. However, the liposomes containing heat-denatured OMP-I were scarcely active. Spontaneous and antigen-specific immunoglobulin M (IgM)-, IgG-, and IgA-secreting spot-forming cells (SFC) enzymatically dissociated into single-cell suspensions from chronically inflamed periapical tissues and were enumerated by enzyme-linked immunospot assay. In patients with radicular cysts or dental granulomas, the major isotype of spontaneous SFC was IgG. In radicular cysts, the OMP-II-specific IgG SFC represented 0.13% of the total IgG SFC, while the antigen-specific IgA or IgM SFC was not observed. It was also found that none of these mononuclear cells produced antibodies specific for OMP-I or lipopolysaccharide of P. endodontalis.

Antibodies, Bacterial

Type A aortic dissection: evaluation with ultrafast CT.

The authors examined 17 patients with type A aortic dissection with ultrafast computed tomography (CT). Forty sections with 1-cm intervals, from the aortic arch to the aortic bifurcation, were scanned serially without breath holding within only 74 seconds. Except for two thrombosed dissections, all intimal flaps in the ascending aorta were demonstrated, and 14 (93%) of the 15 surgically proved intimal tears were depicted. An intimal tear in the aortic arch was missed. Since conventional scanning tends not to depict type A aortic dissection, millisecond-order ultrafast CT scanning has the potential to become the modality of choice for imaging of aortic dissection.

Adult

Aortic aneurysms: growth rates measured with CT.

Because of a lack of information about the rates of growth of aortic aneurysms, such rates in thoracic and abdominal aortic aneurysms were determined. One hundred seventy-one patients with atherosclerotic aortic aneurysm managed nonoperatively were followed up for more than 6 months with sequential computed tomography (CT). There were 211 aneurysms (thoracic aortic, 82; abdominal aortic, 129). The growth rates of thoracic and abdominal aortic aneurysms were 0.42 and 0.28 cm/y, respectively. Aneurysms at the aortic arch (n = 34) grew at a faster average rate (0.56 cm/y) than aneurysms arising at other levels, even when the rate was corrected for the initial diameter. It is recommended that thoracic aortic aneurysms, especially aortic arch aneurysms, be followed frequently with CT examination of size.

Adult

Noncariogenicity of maltitol in specific pathogen-free rats infected with mutans streptococci.

The effect of maltitol on caries development was examined in an experimental caries system employing specific pathogen-free (SPF) Sprague-Dawley rats. Fourteen strains of oral streptococci, including mutans streptococci, did not utilize the maltitol nor produce sufficient acid to demineralize tooth enamel. Furthermore, maltitol did not serve as a substrate for glucosyltransferases of either Streptococcus mutans MT8148R or Streptococcus sobrinus 6715 to synthesize water-insoluble glucan. Maltitol induced no significant dental caries in SPF rats infected with these mutans streptococci, and replacement of the dietary sucrose content with maltitol resulted in a trend towards caries reduction in SPF rats.

Animals

Effects of hyposalivation on the oral microflora of rats fed sucrose or wheat flour diets.

The effects of hyposalivation on the induction of dental caries and on the composition of the oral microflora were examined in specific pathogen-free Sprague-Dawley rats fed either a sucrose or a wheat flour diet with or without inoculation of Streptococcus mutans. Significant dental caries was induced in hyposalivated rats fed diet 2000 containing 56% sucrose, irrespective of infection by S. mutans. Diets containing 56% wheat flour did not induce dental caries in either hyposalivated or sham-operated rats, irrespective of infection by S. mutans. Bacteriological examinations at the end of the experiment demonstrated that the total numbers of lactobacilli and staphylococci increased in hyposalivated rats irrespective of the diet given, while the inoculated stain of S. mutans decreased significantly in hyposalivated rats. These findings suggest that some acidogenic microorganisms such as lactobacilli and staphylococci that can utilize sucrose or glucose but not wheat flour may also promote dental caries in hyposalivated rats.

Animals

Evaluation of myocardium in ischemic heart disease by ultrafast computed tomography.

In this study we evaluated quantitatively the contrast enhancement of the myocardium of 27 patients with infarction and ischemia by ultrafast CT (Imatron C-100) with a scan time of 100 msec. We defined three parameters from the CT numbers of the myocardium and the ventricle in both the early and the late phase (about 4 min) after the injection of non-ionic contrast material; the difference of the CT numbers of myocardium between the early and the late phase (delta CT#), and the ratios of the CT numbers of the myocardium and the ventricular lumen (M/L) in both the early phase and the late phase. The delta CT# for myocardial infarction and severe ischemia were -20 +/- 10HU and -8 +/- 10HU, respectively. These values were significantly lower than the mild ischemic myocardium and normal myocardium (p less than 0.001). The average M/L values for myocardial infarction and ischemia in the early phase were 19 +/- 8% and 16 +/- 6%, respectively. These values were significantly lower than those in normal myocardium (p less than 0.001). The average M/L values for infarction and severe ischemia in the late phase were 90 +/- 18% and 63 +/- 20%, respectively. These M/L were significantly dissociated (p less than 0.001). Our results indicate that with contrast UFCT it is feasible to detect quantitatively the myocardial characteristics of ischemia and infarction.

Adult

Ehlers-Danlos syndrome with soft-tissue contractures.

We report four patients with a form of Ehlers-Danlos syndrome associated with soft-tissue contractures from birth and skin hyperalgesia. In early infancy, these cases were thought to be forms of arthrogryposis multiplex congenita, Larsen's syndrome or Marfan's syndrome. The characteristic triad of signs of Ehlers-Danlos disease gradually appeared from four to six years of age, allowing us to establish the correct diagnosis. We discuss the differential diagnosis of these connective-tissue disorders and the problems of the orthopaedic treatment of the associated joint deformities. Ehlers-Danlos syndrome is a heterogeneous group of metabolic diseases of collagen and our cases constitute a group which is distinct from the conventional types.

Adult