Assay of phylloquinone and menaquinones in human liver.
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Biomedical subjects
Publications and source records attributed to Y Usui.
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The binding properties of Staphylococcus aureus in relation to human platelets were investigated. Protease digestion (pronase E, proteinase K, trypsin), heat treatment (80 degrees C, 30 min), and sonication for 5 min significantly reduced the binding abilities of the staphylococcal cells to 0% (p < .01), 50 +/- 5% (p < .05), and 38 +/- 9% (p < .05), respectively, while mixed glycosidases did not. Inhibition experiments indicated that protein A and various sugars were ineffective. A binding study using biotinylated cell surface fractions extracted from the whole cells of S. aureus indicated that the proteins having apparent molecular weights of 14400 and 16500 estimated by sodium dodecylsulfate-polyacrylamide gel electrophoresis were involved in the binding between S. aureus and human platelets.
For the determination of disodium cromoglycate in urine, a fluorimetric method using HPLC post-column photoirradiation has been developed. The mobile phase consisted of a 35 mmol l-1 phosphate buffer (pH 8)-methanol (7 + 3, %v/v) containing 75 mmol l-1 hydrogen peroxide and 20 mmol l-1 18-crown-6. The 18-crown-6 was used for separation adjustment of the disodium cromoglycate in the urine sample. Photoirradiation was carried out in tubing wound around a germicidal light in a reactor equipped with an air-cooling fan. The fluorescence was monitored with excitation at 325 nm and emission at 448 nm. The calibration graph for disodium cromoglycate was linear over the range 38-2340 ng ml-1 using an injection volume of 100 microliters. The pretreatment of the urine samples consisted of diluting and filtering steps. The mean recovery of disodium cromoglycate from urine was 99.1 +/- 2.4% (n = 6).
We surveyed a stratified random sample of 205 person from 45,500 residents, aged 65 years or older, in a city of Japan using an interview schedule including scales of active-life orientation, social activities and other covariates. A total of 157 (77%) participated in the study. The main results were as follows: 1) Significantly higher scores of active-life orientation were observed in those who currently worked, than in those who did not work in males and females (p < 0.05). In males, scores of active-life orientation were significantly higher in those who had hobbies than in those who did not (p < 0.05); in those who traveled during the past year than in those who did not (p < 0.05). 2) In multiple linear regression analysis, PGC scale scores positively and significantly correlated with score of active orientation in males and females (p < 0.05). Age negatively correlated with the scores in males. Scores of affiliation orientation positively correlated with scores of active orientation, and satisfaction with available social support negatively correlated with the scores in females (p < 0.05).
To clarify the nature of the accumulated lung T cells of pulmonary sarcoidosis, we studied bronchoalveolar and peripheral blood (PB) lymphocytes from Japanese patients for expressed T-cell receptor beta-chain variable (TCR V beta) gene usage. Quantitative polymerase chain reaction technique revealed that the V beta repertoires of bronchoalveolar and PB T cells were heterogeneous. However, the bronchoalveolar lymphocytes from five out of 10 patients showed overrepresentation of T cells, with a few TCR V beta families, which varied among individuals, possibly reflecting a difference in the human leukocyte antigen (HLA) backgrounds. Nucleotide sequence analysis of third complementarity determining region (CDR3) segments in overrepresented V beta families demonstrated limited diversity of the transcripts. Two HLA-DR 12(5)+ individuals with overrepresented V beta 6 gene transcripts in the lungs shared a common amino acid motif within their CDR3. By contrast, non-overrepresented TCR V beta transcripts did not show clonal expansion, and the oligoclonal clones in the lung were not found in PB of the corresponding patient. These results showed that, at least in subsets of Japanese patients with pulmonary sarcoidosis, a limited number of T-cell clones are accumulated in the lung in response to a specific antigen. They may mediate an immune response to shape pathology of the disease.
Der f 2 is a major mite allergen composed of 129 amino acid residues. To determine the major epitopes on Der f 2 recognized by human IgE antibodies, artificial mutations were introduced to Der f 2 protein. The IgE-binding activity of Der f 2 was significantly decreased by deletion of 10 amino acids at the N-terminus or nine amino acids at the C-terminus. Site-directed mutagenesis with a single amino acid replacement by Ala or Leu in both N- and C-terminal regions as well as a central portion was performed to generate 42 single-site mutations. Amino acid replacement around a disulfide bond of Cys8-Cys119 caused a marked decrease in IgE-binding activity. Furthermore, a distinct decrease in IgE-binding was also caused by Ala-substitution close to a disulfide bond of Cys73-Cys78 and by mutations of a few charged residues. From these results, it was concluded that the two disulfide-forming regions of Der f 2 and several charged residues are important for forming major epitope structures recognized by human IgE antibodies.
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A woman aged 62 with long history of chronic relapsing pancreatitis presented with swelling and ulcer in the lower limbs and occasional gastrointestinal bleeding. The radiological imaging showed complete obstruction of Inferior Vena Cava (IVC) at the level of the pancreas and well developed collateral vessels. Portal vein and splenic vein were also obstructed and superior mesenteric venous blood drained into the liver via coronary vein. She was originally found to have pancreas head tumor, which was not resectable. A palliative operation was performed, but histological examination of pancreatic specimen suggested only chronic inflammation and no evidence of malignancy. She was diagnosed as tumor-forming type chronic pancreatitis. Although SPV or SMV-PV obstruction has been recognized as a complication of chronic pancreatitis, IVC obstruction can occur by the same mechanism. This is the only case but one ever reported. Not only splenoportography but IVC-graphy will contribute to more precise understanding of patient's condition with chronic pancreatitis.
Amino acid sequencing of the major mite allergen Der fII purified from mite body extract revealed that it is a mixture of at least two variants. Substitutions were found only at positions in which amino acid variations were predicted from the nucleotide sequences of three cloned cDNAs. When cDNAs corresponding to the three variants were expressed in Escherichia coli, Der fII proteins were produced as inclusion bodies. Denaturation and renaturation with urea converted recombinant Der fII into a protein with three intramolecular disulfide bonds (Cys8-Cys119, Cys21-Cys27, and Cys73-Cys78), which were identical to those previously identified in native Der fII. All the variants, including native Der fII, were equally recognized by human IgE antibodies from 14 different sera of mite-allergic patients. These results suggested that recombinant Der fII proteins assumed a conformation identical to that of the native Der fII and that all the Der fII variants acted as allergens. This result also suggested that IgE antibody binds to the region where there were no amino acid variations. The binding ability of the monoclonal antibody (mAb) 18G8 for the variant clones 1, 2, and 11 Der fII was almost the same. However, mAb 15E11 bound to clone 1 Der fII more efficiently than to clones 2 and 11, whereas mAb 13A4 recognized clone 2 Der fII as the most preferable antigen. This suggested that these antibodies recognized the region of amino acid variations. The stability of Der fII variants was analyzed by measuring their IgE antibody binding activity after heating, freezing, or acid treatment, and was analyzed by resistance to trypsin digestion.(ABSTRACT TRUNCATED AT 250 WORDS)
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We examined the bacteria isolated from urine of patients in our department from 1988 though 1989. A total of 809 strains were isolated from the outpatients. They were 241 strains of gram-positive bacteria (29.8%). E. coli (32.0%) was isolated most frequently, followed by Enterococcus spp. (9.9%), Proteus group (7.4%), S. epidermidis (6.2%), P. aeruginosa (5.1%) and others. A total of 412 strains were isolated from the inpatients. They consisted of 173 strains of gram-positive bacteria (42.0%). Enterococcus spp. (17.0%) was isolated most frequently, followed by P. aeruginosa (12.6%), S. epidermidis (9.7%), E. coli (7.3%), S. aureus (6.6%) and others. Fewer P. aeruginosa, S. marcescens and Proteus group were isolated from the outpatients and fewer S. marcescens, E. coli and K. pneumoniae from inpatients than previously. More S. aureus were isolated abruptly from the inpatients and more E. coli were isolated from the outpatients than previously. E. cloacae, S. epidermidis, S. aureus and Corynebacterium spp. all isolated from inpatients showed lower antimicrobial sensitivity than those isolated from the outpatients.
Significant changes in disk biochemistry were evident in the nuclei pulposi of 25 symptomatic disks that demonstrated abnormal diskography. Compared with the results from ten normal disk from young subjects (mean age, ten years) and from ten asymptomatic disks from adult patients (mean age, 33 years), the symptomatic disks demonstrated significantly greater collagen content than the young and asymptomatic adult subjects, respectively. Significantly fewer total glycosaminoglycans (GAGs) and significantly less water content were also evident for the diseased disks. The mean pH of the nucleus pulposus from 23 patients was also significantly reduced for the symptomatic disks as compared with the mean pH measured for four asymptomatic adult subjects. Although significantly fewer total GAGs were observed for the symptomatic disks, there were no significant differences in the percent composition of hyaluronic acid (HA), chondroitin sulfate (CS), or keratan sulfate (KS) among the normal and symptomatic disks. Significant changes in disk biochemistry are associated with the symptomatic disk, and these changes could contribute to the development of the painful disk syndrome.