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Biomedical subjects

S Kamei

Publications and source records attributed to S Kamei.

At least 37 records · Page 2Linked to original sources
PubMed2000

Association of remnant-like particle cholesterol with coronary artery disease in patients with normal total cholesterol levels.

BACKGROUND: Limited information is available as to whether there is a difference in the association of lipid and fibrinolytic variables with coronary artery disease according to the presence or absence of elevated serum total cholesterol. We examined the levels of various lipid and fibrinolytic variables including remnant-like particle cholesterol (RLP-C). RLP-C is a recently established simple assay method for the estimation of triglyceride-rich lipoprotein remnants. METHODS AND RESULTS: Levels of total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol (HDL-C), triglyceride, lipoprotein(a), RLP-C, uric acid, blood glucose, tissue plasminogen activator, tissue plasminogen activator inhibitor type 1, antithrombin III, and protein C were measured in 208 patients who underwent diagnostic coronary angiograms. Of these 208 patients, 57 were hypercholesterolemic (> or =220 mg/dL) and 151 were normocholesterolemic. HDL-C showed significant differences between patients with and those without angiographically determined coronary artery stenosis in both hypercholesterolemic and normocholesterolemic patients (P =.0025 and P =.0003, respectively). Both RLP-C and uric acid showed significant differences only in the normocholesterolemic subgroup (P =.0006 and P =.0060, respectively). This difference in uric acid was not significant by multivariable analysis. The ratio of RLP-C/HDL-C was demonstrated to be highly significantly (P <.0001) associated with coronary artery stenosis in patients with normal total cholesterol, whereas there was no statistically significant association in the hypercholesterolemic patient subgroup. CONCLUSIONS: Our current study disclosed that RLP-C levels are strongly associated with coronary artery disease, especially in patients with normal total cholesterol levels. Moreover, RLP-C/HDL-C ratio may be even more significantly associated with the presence of coronary artery stenosis in normocholesterolemic patients.

Adult↗
PubMed2000

Predictive value of remnant-like particle cholesterol as an indicator of coronary artery stenosis in patients with normal serum triglyceride levels.

OBJECTIVE: We designed the present study to evaluate the association of various lipid and fibrinolytic components with coronary artery stenosis with respect to the triglyceride (TG) level. METHODS: Levels of TG, remnant-like particle cholesterol (RLP-C), total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), lipoprotein-(a), uric acid, blood glucose, tissue plasminogen activator (t-PA), t-PA inhibitor type 1, antithrombin III, and protein C were measured in 208 patients who underwent diagnostic coronary angiograms. PATIENTS: Of these 208 patients, 59 were hypertriglyceridemic (150 mg/dl or higher) and 149 were normotriglyceridemic. RESULTS: Both LDL-C and HDL-C showed significant differences between patients with and those without coronary artery stenosis in both hypertriglyceridemic and normotriglyceridemic patient subgroups. However, RLP-C showed a significant difference in the normotriglyceridemic patient subgroup (p=0.012) but not in the hypertriglyceridemic patient subgroup (p=0.736). CONCLUSION: Our current retrospective study disclosed that RLP-C levels are closely associated with coronary artery stenosis in patients with normal TG levels.

Aged↗
PubMed2000

Nationwide survey of the annual prevalence of viral and other neurological infections in Japanese inpatients.

OBJECTIVE: To estimate the annual prevalence of viral and other neurological infections at large hospitals in Japan during the period from 1989 to 1991. METHODS: A nationwide questionnaire survey on the numbers of inpatients with viral and other neurological infections was sent for completion to the chiefs of Departments of Internal Medicine, Neurology and Pediatrics at all hospitals with more than 200 beds. RESULTS: The average annual number of inpatients (and the number per 10(6) population) with encephalitis in large hospitals was estimated to be 2,200+/-400 (17.7+/-3.2), while it was 32,000+/-16,000 (258+/-129) for meningitis, and 650+/-50 (5.2+/-0.4) for myelitis. Among the inpatients with encephalitis, meningitis, and myelitis, an unknown etiology was the most common (51.2% in encephalitis, 73.2% in meningitis, and 36.3% in myelitis), followed by a viral etiology for all three diseases. CONCLUSION: The first estimate was made of the annual prevalence of viral and other neurological infections and their etiology in Japan.

Adult↗
PubMed2000

Nucleotide sequence of the gene encoding murine tissue factor pathway inhibitor-2.

Tissue factor pathway inhibitor-2 (TFPI-2), also known as placental protein 5, is a 32 kDa extracellular matrix-associated serine proteinase inhibitor consisting of three tandemly-arranged Kunitz-type domains. Two overlapping genomic clones containing sequences encoding murine TFPI-2 were isolated from a lambda FIXII 129 SVJ mouse genomic library, and the complete nucleotide sequence of the gene was determined. The murine TFPI-2 gene spans approximately 9.3 kilobases and consists of five exons and four introns. The nucleotide sequences surrounding all the exon-intron boundaries are highly conserved and obey the GT-AG rule. Each Kunitz-type domain is encoded by a single exon, similar to that observed for other Kunitz-type proteinase inhibitors. A total of 1,577 bp of the 3'-flanking region contains a probable polyadenylation site (ATTAAA) at +5,759 and an apparent cleavage or termination site (CATTG) at +6,170. The 5'-flanking region of the murine TFPI-2 gene contains a prototypical TATA box, a GC box and two CAAT boxes. In addition, several candidate transcription factor binding sites responsible for placenta-, endothelial cell-, and smooth muscle cell-specific expression of the TFPI-2 gene were also identified.

Amino Acid Sequence↗
PubMed2000

[Sarcomatoid carcinoma of the urinary bladder: report of a case].

The patient was a 78-year-old man with gross hematuria. A non-papillary and non-pedunculated tumor was found on the posterior wall of the bladder by cystoscopic examination. Total cystectomy with construction of ileal conduit was performed. Histologically, the tumor was composed of carcinomatous and sarcomatous elements. The carcinomatous element was compatible with grade 3 transitional cell carcinoma. The sarcomatous element was composed of osteosarcomatous, chondrosarcomatous pattern and undifferentiated malignant spindle cell component. Immunohistochemical examination demonstrated the presence of keratin, cytokeratin and epithelial membrane antigen in both carcinomatous and sarcomatous elements. Therefore, we diagnosed this tumor as sarcomatoid carcinoma. The patient has remained well without any evidence of recurrence for 10 months after operation.

Aged↗
PubMed2000

[Small cell carcinoma of the urinary bladder: a case report].

We report a case of small cell carcinoma of the urinary bladder. The patient was a 58-year-old man complaining of gross hematuria. Clinical examination revealed a non-papillary, broad-based tumor on the right bladder wall with a clinical stage of T3a, N0, M0. Neoadjuvant intraarterial infusion chemotherapy with methotrexate, adriamycin and cisplatin was performed, but it was ineffective. Three weeks later we performed a radical cystectomy. The operative specimen of the tumor revealed small cell carcinoma. It was staged pT3aN0M0R0L2V1. Postoperatively, 1 course of adjuvant chemotherapy using cisplatin and etoposide was performed. The patient is alive without any evidence of tumor recurrence at 6 months after operation.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2000

Physical aging by soft ultrasonic wave enhances ethanol metabolism: metabolic process of wine as followed by 400 mhz 1H-NMR spectroscopy.

In natural aging of spirits or wine, the dynamic structure of ethanol-water clusters changes to a smaller and more uniform state. Through experience we know that naturally aged ones have higher metabolism than the non-aged ones. Also, the same effect as natural aging can be obtained in various types of spirits or wines by the treatment for a period of time with soft ultrasonic wave (US). In this study, we compared ethanol metabolism in human subjects dosed with non-treated white wine (control = CON) and with US treated wine. Ethanol levels in human sera were followed by 400 MHz 1H-NMR spectroscopy after administration of wine doses. Experimental results indicated that ethanol metabolism was enhanced 18% in subjects when US treated wine was used rather than when non-treated (CON) was used. Other experiments using rabbits showed that a 20% ethanol-aqueous solution was absorbed 18% more rapidly by the group dosed with US wine than by the CON group. From these experimental facts, it was theorized that ethanol metabolism depends on the rapidity of ethanol absorption in the human body. And it can be concluded that US treatment brings about the same effect on spirits or wines as natural aging.

Animals↗
PubMed2000

Serum concentration of interferon-alpha: a comparison between once-a-day and twice-a-day administration.

The aim of this study was to determine the possible mechanism of the antiviral activity of twice daily administration of interferon (IFN)-alpha for active hepatitis C and to evaluate serum concentrations of IFN-alpha with enzyme-linked immunosorbent assay (ELISA). Forty-seven patients with active hepatitis C received IFN-alpha intramuscularly for 24 weeks. They were divided into once-a-day and twice-a-day groups. Changes in serum IFN-alpha levels were assessed during the treatment period. Compared with twice-a-day treatment, the once-a-day group showed no increase in serum IFN-alpha at the end of daily treatment (p < 0.03). In contrast, one-third of the twice-a-day group showed increased IFN-alpha at the end of daily treatment (p < 0.02). In conclusion, measured with ELISA, twice-daily administration of IFN-alpha manifested prolonged elevation in serum levels when compared with once daily administration.

Adult↗
PubMed1999

Stable expression of protective protein/cathepsin A-green fluorescent protein fusion genes in a fibroblastic cell line from a galactosialidosis patient. Model system for revealing the intracellular transport of normal and mutated lysosomal enzymes.

Fibroblastic cell lines derived from a galactosialidosis patient, stably expressing the chimaeric green fluorescent protein variant (EGFP) gene fused to the wild-type and mutant human lysosomal protective protein/cathepsin A (PPCA) cDNA, were first established as a model system for revealing the sorting and processing of lysosomal enzymes and for investigating the molecular bases of their deficiencies. In the cell line expressing the wild-type PPCA-EGFP chimaera gene (EGFP-PPwild), an 81 kDa form (27 kDa EGFP fused to the C-terminus of the 54 kDa PPCA precursor) was produced, then processed into the mature 32/20 kDa two-chain form free of the EGFP domain. The intracellular cathepsin A, alpha-N-acetylneuraminidase and beta-galactosidase activities, which are deficient in the parent fibroblastic cells, could also be significantly restored in the cells. In contrast with the uniform and strong fluorescence throughout the cytoplasm and nucleus in the mock-cell line expressing only EGFP cDNA, weak reticular and punctate fluorescence was distributed throughout the EGFP-PPwild cell line. Bafilomycin A1, a potent inhibitor of vacuolar ATPase and intracellular acidification, induced the distribution of Golgi-like perinuclear fluorescence throughout the living and fixed cells, in which only the 81 kDa product was detected. After removal of the agent, time-dependent transport of the chimaeric protein from the Golgi apparatus to the prelysosomal structure in living cells was monitored with a confocal laser scanning microscope system. Leupeptin caused the distribution of lysosome-like granular fluorescence throughout the cytoplasm in the fixed cells, although it was hardly observed in living cells. The latter agent also dose-dependently induced an increase in the intracellular amount of the 81 kDa product containing the EGFP domain and inhibited the restoration of cathepsin A activity in the EGFP-PPwild cells after the removal of bafilomycin A1. In parallel, both the mature two-chain form and PPCA function disappeared. These results suggested that the chimaera gene product was transported to acidic compartments (endosomes/lysosomes), where proteolytic processing of the PPCA precursor/zymogen, quenching of the fluorescence, and random degradation of the EGFP portion occurred. A cell line stably expressing a chimaeric gene with a mutant PPCA cDNA containing an A1184-->G (Y395C) mutation, commonly detected in Japanese severe early-infantile type of galactosialidosis patients, showed an endoplasmic reticulum (ER)-like reticular fluorescence pattern. The PPCA-immunoreactive gene product was hardly detected in this cell line. The mutant chimaeric product was suggested to be degraded rapidly in the ER before transport to post-ER compartments. A cell line expressing the chimaeric gene with a T746-->A (Y249N) PPCA mutation exhibited both ER-like reticular and granular fluorescence on the reticular structure that was stronger than that in the EGFP-PPwild cells. Some of them contained large fluorescent inclusion-body-like structures. The ineffectiveness of transport inhibitors in the distribution changes in the two mutant chimaeric proteins suggested that they were not delivered to acidic compartments. Therefore this expression system can possibly be applied to the direct analysis of the sorting defects of mutant gene products in living cells and will be useful for the molecular investigation of lysosomal diseases, including galactosialidosis.

Amino Acid Sequence↗
PubMed1999

Inhibitory properties of human recombinant Arg24-->Gln type-2 tissue factor pathway inhibitor (R24Q TFPI-2).

Human type-2 tissue factor pathway inhibitor (TFPI-2), also known as placental protein 5, is a 32-kDa serine proteinase inhibitor consisting of three tandemly arranged Kunitz-type domains homologous to tissue factor pathway inhibitor. TFPI-2 inhibits a variety of serine proteinases involved in coagulation and fibrinolysis through an arginine residue (R24) in its first Kunitz-type domain, which constitutes a putative P1 residue for the substrate recognition sites of these proteinases. As recent studies have shown that this P1 residue to be a glutamine in murine TFPI-2, we constructed, expressed, and purified a human TFPI-2 mutant with glutamine substituted for arginine at position 24 (R24Q TFPI-2). R24Q TFPI-2 lost approximately 90% of its inhibitory activity towards bovine trypsin and virtually all inhibitory activity towards human plasmin and the factor VIIa-tissue factor complex, emphasizing the importance of the P1 Arg24 residue in the inhibition of these serine proteinases. However, whereas wild-type TFPI-2 is a relatively weak inhibitor of human factor Xa amidolytic activity (IC50 approximately 1 microM), R24Q TFPI-2 exhibited enhanced inhibitory activity towards the amidolytic and coagulant activities of this proteinase with a Ki of 18 nM. While the molecular basis for the enhanced inhibition of human factor Xa by R24Q TFPI-2 is unknown, these data provide suggestive evidence that murine TFPI-2 may function as a serine proteinase inhibitor in spite of the absence of a P1 Arg or Lys residue.

Amino Acid Sequence↗
PubMed1999

Simultaneous analysis of serum immunoglobulins in patients with M protein using cellulose acetate membrane isoelectric focusing.

We developed a method for the simultaneous analysis of microheterogeneity of human serum IgG, IgA, IgM, IgD, and IgE, and serum protein pattern using cellulose acetate membrane isoelectric focusing, and analyzed in 11 healthy subjects and 67 patients with M protein (17 cases of multiple myeloma [MM] and 50 cases of monoclonal gammopathy of undetermined significance [MGUS]). Using this method, bands indicating the microheterogeneity of each immunoglobulin could clearly be detected.Among healthy subjects, the detected IgG, IgA, and IgM bands did not vary, but the detected IgE and IgD bands did vary. Therefore, IgA, IgM, and IgG were selected for comparison of serum immunoglobulins in MM and in MGUS. In the IgA-type M protein group, normal IgM and IgG bands were decreased in MM patients compared to MGUS patients, while the M band and other bands were increased in MM patients compared to MGUS patients, but the differences between the two groups were not significant. In the IgG-type M protein group, normal IgM, IgA, and IgG were significantly decreased in MM patients compared to MGUS patients. We examined the changes in electrophoretic pattern in six MM patients and eight MGUS patients with IgA-type M protein after neuraminidase treatment. The width of the M band in MM patients with IgA-type M protein decreased with neuraminidase treatment. On the other hand, the width of the M band in MGUS patients with IgA-type M protein increased with neuraminidase treatment. We concluded that the decrease of the normal immunoglobulins in MM patients with IgG type M protein could be detected by this method, and IgA type of M protein binding sugar chain were different between MM and MGUS patients.

Blood Proteins↗
PubMed1999

Blinded, prospective, and serial evaluation by quantitative-EEG in interferon-alpha-treated hepatitis-C.

OBJECTIVES: To estimate the effect of brain function due to IFN-alpha in chronic hepatitis C patients by the quantitative EEG. METHODS: 56 chronic hepatitis C patients were administered IFN-alpha intramuscularly at 9x10(6) IU daily for the first 4 weeks and then 3 times/week for the next 20 weeks. Serial EEGs were obtained in each subject before, at 2 and 4 weeks, and after completing the treatment. Resting EEG without artifacts was selected for quantitative EEG analysis, which was performed blindly. The frequency range was divided into delta to beta. The absolute and relative powers of each frequency band in each subject were calculated and the differences of these powers at different stages were analyzed statistically. RESULTS: The absolute powers of slow waves (delta, theta 1, and theta 2) increased while alpha 2 and fast wave (beta) decreased significantly at all locations during IFN-alpha administration. The total power and alpha 1 values revealed no significant alterations. The relative power revealed the same alteration during treatment. These changes disappeared following the treatment. Such diffuse slowing in the EEG was revealed by the total change in the whole subjects. CONCLUSIONS: Diffuse slowing in the EEG was induced by IFN-alpha, was reversible, and was evident as the total change in the subjects. These findings suggested mild IFN-alpha-induced encephalopathy.

Analysis of Variance↗
PubMed1999

Comparative study between chemiluminescence assay and two different sensitive polymerase chain reactions on the diagnosis of serial herpes simplex virus encephalitis.

OBJECTIVE: A prospective study was undertaken on the diagnosis of herpes simplex encephalitis (HSVE) by comparing chemiluminescence assay (CL) and two different sensitive polymerase chain reactions (PCRs). METHODS: The materials comprised 53 serial CSF samples from 31 patients with acute encephalitis with suspected HSVE. Each CSF was distributed to three independent laboratories to perform quantitative measurements by CL, the low sensitive (single) PCR, and high sensitive (nested) PCR. The CL provided a method of detecting HSV itself and the small fragment with HSV antigenicity which was composed of viral component proteins. The serial CSFs were found retrospectively to comprise 24 samples from 11 patients with HSVE due to HSV1 and 29 samples from 20 patients with non-HSVE. RESULTS: the CL showed 50 to 48 000 pfu/ml in all samples of HSVE (except one) taken from the 3rd to the 25th day. The low sensitive PCR demonstrated 50 to 47 000 pfu/ml in only six samples of HSVE. The high sensitive PCR disclosed less than 100 to 120 000 copies/ml in 11 samples of HSVE. At the acute stage from the 1st to 7th day, the sensitivities of CL and the high sensitive PCR were 100%, but that of the low sensitive PCR was 75%. The sensitivity of CL was significantly higher than those of both PCRs after the acute stage on the 15th to 32nd day. The specificities and positive predictive values of the three methods were 100%. However, the negative predictive value of CL was significantly higher than that of the low sensitive PCR. CONCLUSIONS: The sensitivity of CL is equivalent to that of the high sensitive PCR during the acute stage and significantly higher than that of the high sensitive PCR after the acute stage. A clear difference in sensitivity exists between the different PCRs. A combination of the PCR, chemiluminescence assay, and serological antibody diagnosis is currently considered the most effective approach for the clinical diagnosis of HSVE.

Adult↗
PubMed1998

Stabilizing effect of lysosomal beta-galactosidase on the catalytic activity of protective protein/cathepsin A secreted by human platelets.

The 32/20-kDa two-chain form of protective protein/cathepsin A (CathA) secreted by human platelets was thermostable in the aggregation supernatant at acidic pH, but was denatured at neutral pH. Leupeptin partly protected the CathA against denaturation, which was not observed in the supernatant after depletion of the cosecreted lysosomal acid beta-galactosidase (beta-Gal) by affinity separation with p-aminophenylthiogalactose (PATG)-agarose beads even at pH 4.8. The purified recombinant human beta-Gal proteins, the 84-kDa precursor and 64-kDa mature-like enzyme (the tryptic product of the 84-kDa precursor), also protected the CathA against denaturation at neutral pH in part. Biospecific interaction analysis revealed that the CathA secreted by platelets dose dependently bound to the immobilized recombinant beta-Gal proteins. The association rate constant of CathA with the 64-kDa mature-like beta-Gal was 4.0 x 10(6) (M-1 s-1) at acidic pH, which was three times larger than that with the 84-kDa beta-Gal precursor. The calculated affinity constants for the enzyme molecules at acidic pH were approximately 1 x 10(9) (M-1), and those at neutral pH were two orders lower. These results first demonstrated that beta-Gal stabilizes the catalytic activity of CathA through direct binding in vitro. The affinity was shown to increase with removal of the carboxy-terminal domain of the beta-Gal precursor.

Blood Platelets↗
PubMed1998

A kinetic analysis of the interaction of human recombinant tissue factor pathway inhibitor with factor Xa utilizing and immunoassay and the effect of antithrombin III/heparin on the complex formation.

We have recently shown that a complex formation of tissue factor pathway inhibitor (TFPI) and factor Xa (Xa) promotes a clearance of proteoglycans-associated TFPI. In the current studies, the interaction between human recombinant TFPI (h-rTFPI) and Xa were kinetically analyzed by utilizing both a protease inhibitor, p-(amidophenyl) methanesulfonyl fluoride hydrochloride, and a specific enzyme-linked immunosorbent assay for the complex of h-rTFPI with Xa. We further investigated the effect of antithrombin III on the complex formation between h-rTFPI and Xa. We found that the h-rTFPI/Xa complex formed in a time-dependent manner: the second-order rate constant (K1) for the complex formation was calculated to be 0.86x10(6) M(-1)s(-1). The addition of antithrombin III to the h-rTFPI solution modestly reduced the rate of the complex formation between h-rTFPI and Xa. Heparin strikingly enhanced antithrombin III's inhibition of Xa and resulted in complete abrogation of the complex formation between h-rTFPI and Xa in the absence or presence of acidic phospholipids. Furthermore, antithrombin III induced dissociation of the preformed h-rTFPI/Xa complex in the presence of heparin. These results suggest that in the presence of heparin, antithrombin III interferes with the catabolism of TFPI mediated via Xa.

Antithrombin III↗
PubMed1998

Predictive value of remnant-like particles cholesterol/high-density lipoprotein cholesterol ratio as a new indicator of coronary artery disease.

BACKGROUND: There is as yet no definite consensus on the predictive value of the various lipid profiles and fibrinolytic parameters that became available in clinical use recently for coronary artery disease. METHODS: Levels of lipoprotein(a), high-density lipoprotein cholesterol (HDL-C), remnant-like particles cholesterol (RLP-C), tissue plasminogen activator (TPA), TPA inhibitor, antithrombin III, and protein C were measured in 124 patients who underwent diagnostic coronary angiograms. RESULTS: Of these patients, 37 had no significant stenoses (group N) and 87 had significant stenoses (group S). There were no significant differences in patient characteristics between the two groups. HDL-C was significantly lower (p = 0.0071 ) and RLP-C was significantly higher (p = 0.0022) in group S. When a product and a ratio of each of two factors were calculated, RLP-C/HDL-C was demonstrated to be a highly significant predictor for coronary artery stenoses (p < 0.0001). There were also significant increases in RLP-C/HDL-C levels with increasing number of vessels involved (r = 0.359, p < 0.0001 ). CONCLUSION: Our present study disclosed the predictive value of RLP-C/HDL-C ratio as a new indicator of coronary artery disease.

Adult↗
PubMed1998

Development of a sensitive and accurate enzyme-linked immunosorbent assay (ELISA) system that can replace HPLC analysis for the determination of N1,N12-diacetylspermine in human urine.

N1,N12-Diacetylspermine (DiAcSpm)-specific antibodies were raised in rabbits, using N-acetylspermine coupled to mercaptosuccinylated BSA via N-(4-maleimidobutyryloxy)-succinimide as an antigen. Highly DiAcSpm-specific antibodies were enriched from crude sera through a series of affinity-based fractionations. A competitive ELISA system, intended for measuring DiAcSpm in solution, was constructed using this antibody preparation, with N-acetylspermine coupled to a synthetic peptide via N-(8-maleimidocapryloxy)-succinimide as a solid phase antigen. The Ki value for DiAcSpm with this competitive ELISA system was 33 nM, and the cross-reactivity with DiAcSpm, AcSpm, DiAcSpd, N1-AcSpd, and N8-AcSpd was 100, 0.29, 0.20, 0.033, and 0.055%, respectively. This procedure can be applied to the determination of DiAcSpm in human urine samples, giving highly reproducible results. The coefficients of variation obtained were 6.7 and 4.2% for within-run and between-run precision, respectively. The correlation coefficient between DiAcSpm concentrations in urine estimated by ELISA and those by HPLC analysis was calculated to be 0. 99, and the regression equation was expressed as y = 1.04x + 0.026 microM.

Amino Acid Sequence↗