Biomedical subjects

Y Tsuruta

Publications and source records attributed to Y Tsuruta.

At least 55 records · Page 3Linked to original sources

PubMed1998

Recognition of rheumatoid arthritis synovial antigen by CD4+,CD8- T cell clones established from rheumatoid arthritis joints.

OBJECTIVE: To investigate the rheumatoid arthritis (RA)-specific autoantigen(s) recognized by CD4+ T cells in patients with RA. METHODS: CD4+,CD45RO+ T cell clones were established from the joints of RA patients, and were examined for their proliferative response to synovial cells. RESULTS: Eight of 146 T cell clones responded to RA synovial cells in a DR-restricted manner. These T cell clones recognized solubilized antigens extracted from RA synovial cells in the presence of DR-matched antigen-presenting cells, but did not respond to those extracted from non-RA synovial cells. The antigens had a molecular weight of 50/25 kd. Five of the 8 T cell clones used T cell receptor BV6, and the remaining clones used BV12.2. CONCLUSION: The antigens recognized by joint-infiltrating CD4+ T cells are present exclusively in RA synovial cells. The expression of these antigens by synovial cells may trigger the autoreactivity of T cells in RA joints.

Amino Acid Sequence↗

PubMed1998

Tumor response to neoadjuvant chemotherapy correlates with the expression of P-glycoprotein and PCNA but not GST-pi in the tumor cells of cervical carcinoma.

OBJECTIVE: To identify the clinicopathological and chemoresistant factors predicting the response to neoadjuvant chemotherapy and the patient prognosis in high-risk cervical carcinomas. METHODS: We retrospectively reviewed 47 patients with locally advanced or bulky cervical carcinoma treated with two courses of intraarterial infusion of cisplatin, doxorubicin, mitomycin C, and 5-fluorouracil (5-FU), followed by radical hysterectomy at our hospital between 1988 and 1995. Expressions of the chemoresistance-related proteins, such as P-glycoprotein, glutathione S-transferase pi (GST-pi), and proliferating cell nuclear antigen (PCNA) in the tumor cells, were examined by immunohistochemistry using pretreatment biopsy specimens. These results were compared with the chemotherapeutic response, which was evaluated by magnetic resonance imaging (MRI) and histopathology. Outcome of the patients was also studied. RESULTS: Chemotherapeutic effect of either complete (CR) or partial (PR) response on MRI was obtained in 36 of the 47 (86%) patients. Poor response to chemotherapy was significantly correlated with P-glycoprotein expression (P < 0.005) and low PCNA labeling (P < 0. 05), but not GST-pi expression in the tumor cells. Independent prognostic factors for patient survival were parametrial involvement and lymph node metastasis. Neither the expression of GST-pi nor PCNA was correlated with the patient survival. CONCLUSION: Assessment of the expression of P-glycoprotein and PCNA is potentially useful for the prediction of tumor response to neoadjuvant chemotherapy for cervical carcinomas.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1998

Amyloid arthropathy of the hip joint: MR demonstration of presumed amyloid lesions in 152 patients with long-term hemodialysis.

The aim of this study was to determine the spectrum of MR findings of presumed amyloid arthropathy of the hip joints in patients on long-term hemodialysis. We prospectively performed T1- and T2-weighted spin-echo imaging on 152 consecutive patients on hemodialysis. The duration of hemodialysis ranged from 5 months to 24 years, 2 months (mean: 8 years, 8 months). The frequency, location, and signal intensity of bone lesions were assessed. In 12 cases with contrast-enhanced MR examination, enhancement pattern of bone lesions, synovial lesions, and intra-articular lesions were characterized. Bone lesions presumed to be amyloid deposits were identified in 60 patients (39%). Magnetic resonance imaging revealed that amyloid lesions were more extensive than anticipated by plain radiographs. All bone lesions showed decreased signal intensity on T1-weighted images. On T2-weighted images, bone lesions showed increased signal intensity in 32 patients (54%), decreased signal intensity in 11 patients (18%), and both increased and decreased signal intensity in 17 patients (28%). Following intravenous injection of gadolinium-based contrast, all bone lesions showed moderate enhancement. Synovial thickening could not be identified on T1- and T2-weighted images. However, contrast-enhanced images showed thickened synovial membrane, which could be differentiated from joint fluid. Intra-articular nodules showed decreased or intermediate signal intensity on T1-weighted images and decreased signal intensity on T2-weighted images; the intra-articular nodules were contiguous with subchondral bone lesions. Magnetic resonance imaging is useful for evaluating the distribution and extent of amyloidosis of the hip joints in patients undergoing long-term hemodialysis.

Adult↗

PubMed1998

Heteroduplex analysis: a useful screening method for glycogen storage disease type Ia.

Glycogen storage disease type Ia (GSDIa), also known as von Gierke disease, is the most common and severe disease of glycogenoses and is caused by a deficiency of glucose-6-phosphatase (G6Pase) and transmitted by an autosomal recessive trait. The encoding gene of G6Pase is composed of only five exons and each exon is short. With heteroduplex analysis (HDA) method, we analyzed the genomic DNA from a patient diagnosed with GSDIa and from her parents. Exons II and IV of the patient showed heteroduplex bands. The mother had a heteroduplex band of exon II, and the father had a heteroduplex band of exon IV. In a mini-slab electrophoresis, exons II and IV of the patient did not show clear heteroduplex bands, but they appeared broader than the others, which made us suspect that they were heteroduplex bands. HDA is an easy and simple method and can verify mutant homozygous DNA fragments by adding wild-type DNA. We think that HDA may be a very useful screening method for the detection of novel genomic mutation in GSDIa in large-scale and mini-slab electrophoresis.

Adult↗

PubMed1998

Early invasive adenocarcinoma of the fallopian tube: a case report and review of the literature.

We present an early invasive adenocarcinoma of the fallopian tube, which was incidentally found in a 45-year-old woman undergoing a laparotomy for uterine myoma. Histological examination of the hydropic tubes revealed widespread endosalpingeal hyperplasia without atypia in both tubes. In addition, the left tube contained 3 scattered lesions of carcinoma in situ, one of which was accompanied by a microfocus of definite stromal invasion confined within the endosalpingeal mucosa. Such a case seems extremely rare, and it might represent the histological appearance of an early invasive feature of tubal carcinoma. We reviewed previously reported cases of in situ and/or early invasive carcinomas of the fallopian tube with respect to the pathological diagnosis and histogenesis of primary tubal adenocarcinomas.

Adenocarcinoma↗

PubMed1998

Lipid peroxidation induced by adriamycin in linolenic acid-loaded cultured hepatocytes.

Addition of more than 10 microM of adriamycin to cultured rat hepatocytes loaded with alpha-linolenic acid (linolenic acid-loaded hepatocytes) caused marked lipid peroxidation as measured by an accumulation of malondialdehyde during a 9 hr incubation. After addition of 50 microM of adriamycin to linolenic acid-loaded hepatocytes, malondialdehyde accumulation significantly increased at 3 hr, followed by cellular reduced glutathione decrease and lactate dehydrogenase leakage after 6 hr. Inhibition of adriamycin-induced lipid peroxidation by addition of N,N'-diphenyl-p-phenylenediamine or alpha-tocopherol, both lipid radical scavengers, or deferoxamine, which is a Fe ion chelator, prevented both glutathione decrease and lactate dehydrogenase leakage, indicating that lipid peroxidation caused cellular damage to linolenic acid-loaded hepatocytes exposed to adriamycin. The effect of SKF 525-A, which is a cytochrome P450 inhibitor, on adriamycin-induced lipid peroxidation and on 7-ethoxycoumarin O-deethylase activity was determined by 6 hr incubation of linolenic acid-loaded cells. Addition of SKF 525-A suppressed adriamycin-induced lipid peroxidation comparably with its 7-ethoxy-coumarin 0-deethylase inhibitory activity. These results suggest that cytochrome P450 contributes to the one-electron bioreduction of adriamycin into its semiquinone radical in rat hepatocytes.

7-Alkoxycoumarin O-Dealkylase↗

PubMed1998

Effect of potassium sorbate on cellular GSH level and lipid peroxidation in cultured rat hepatocytes.

Change in cellular reduced glutathione (GSH) level was examined after the addition of 1-10 mM potassium sorbate (SA-K) to cultured rat hepatocytes. The cellular GSH content was decreased to the lowest level at 6 h after the addition of SA-K, and then gradually returned to the normal level except for hepatocytes exposed to 10 mM SA-K. Although the decrease in GSH level was not associated with lactate dehydrogenase (LDH) leakage in hepatocytes exposed to SA-K up to the concentration of 5 mM, cell injury was caused in cells exposed to 10 mM SA-K. When eicosapentaenoic acid was added in conjunction with various concentrations of SA-K to hepatocytes, peroxidation of the fatty acid was accelerated in parallel with the decrease in cellular GSH level. The enhanced lipid peroxidation in the hepatocytes co-exposed to SA-K and eicosapentaenoic acid (EPA) induced the development of cell injury. These results suggest that hepatocytes exposed to SA-K become susceptible to oxidative stress such as lipid peroxidation.

Animals↗

PubMed1998

[Flow cytometric analysis of proliferative activity of pleomorphic adenoma of salivary gland].

Pleomorphic adenoma (PA) of the salivary gland has diverse biological behavior in spite of its being a benign tumor. So the nuclear DNA content of 36 PAs was measured by flow cytometry to determine the relationship between proliferative activity and histopathological variable. DNA histograms were evaluated according to the rate of S and G2+ M phase cells (S + G2M%). We assumed that the DNA histogram measured algebraically, S + G2 M% < or = 0%, is the near diploid pattern and calculated its ratio to the near diploid pattern. A statistically significant difference between PA and normal salivary gland was found by the ratio to the near diploid pattern (P < 0.05), which confirmed the usefulness of the ratio to the near diploid pattern for measuring low proliferative activity. The tumors were divided into 3 groups: epitheloid type, intermediate type and myxochondroid type. In the 3 groups no differences were found by S + G2M% and the ratio to the near diploid pattern. The areas of the tumor were divided into 5 groups according to the ratio in the epitheloid region and the myxochondroid region. In the 5 groups, the area which consists partly of epitheloid components and mostly of myxochondroid components had the lowest ratio to the near diploid pattern and the highest S + G2M%, and the area which consists of only myxochondroid components had the highest ratio to the near diploid pattern and the lowest S + G2M%. Between only these two areas a statistically significant difference was found by the ratio to the near diploid pattern (P < 0.05). We considered that the area which consists partly of epitheloid components, and mostly of myxochondroid components has the highest proliferative activity in PA. Neither aneuploid or polyploid cells were found in any tumor, but the S + G2M% is more than 20% in 4 tumors. None of these high S + G2M% tumors except one recurrent tumor had clinical and histopathological features. A differences between PAs of the parotid glands and PAs of the submandibular glands was not found in S + G2M%.

Adenoma, Pleomorphic↗

PubMed1997

Quantitative analysis of the usage of human T cell receptor alpha and beta chain variable regions by reverse dot blot hybridization.

We previously developed an adaptor ligation-mediated PCR method to amplify the T cell receptor (TCR) cDNA pools. In the present study we applied reverse dot blot hybridization to PCR-amplified specimens for quantitative analysis of the usage of TCR alpha and beta chain variable (V) region. 44 VA sequence-specific oligonucleotide probes (SSOPs) and 38 VB SSOPs were synthesized corresponding to unique sequences of VA and VB subfamilies. Peripheral blood lymphocytes of ten healthy donors and five T cell clones established from bone marrow cells were examined for VA and VB usage using this method. The results were consistent with those obtained by a colony hybridization method and those by immunofluorescence staining using monoclonal antibodies to VA and VB. Thus, reverse dot blot hybridization for TCR V(alpha) and Vbeta is a new, easy and dependable technique useful for analysis of VA and VB usage by human T cells.

Base Sequence↗

PubMed1997

Non-restricted T cell receptor (TCR)-V alpha and -V beta gene usage in patients with pulmonary sarcoidosis.

Sarcoidosis is a systemic granulomatous disease of unknown etiology characterized by the pronounced accumulation of CD4+ T cells and macrophages in the affected organs. TCR variable (V) alpha and V beta gene usage in patients with sarcoidosis is still a matter of discussion. In this investigation, we analysed TCR-V alpha and -V beta gene usage in bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cells (PBMC) of 30 patients with active pulmonary sarcoidosis using an adapter ligation method, reverse transcriptase-polymerase chain reaction (RT-PCR), and sequence-specific oligonucleotide probe (SSOP) analyses. There was no significant difference in TCR-V alpha or -V beta gene usage between BALF (n = 12) or PBMC (n = 27) of patients and PBMC of healthy subjects (n = 10). Neither selective TCR-V alpha nor -V beta expansion was observed in the paired BALF and PBMC from seven of nine patients. However, selective expansions were observed in a few TCR-V alpha or -V beta subsets in the BALF or PBMC of some individuals. Although a modest increase in a few TCR-V alpha or -V beta subsets was observed in the BALF or PBMC of some individuals, the increased TCR-V alpha or -V beta subsets were not closely associated with the HLA-DRB1, DQA1, DQB1, and DPB1 alleles of these patients. These results suggest that TCR-V alpha or -V beta gene usage is not restricted in both lung and peripheral blood in the majority of patients with active pulmonary sarcoidosis.

Adult↗

PubMed1997

Expression of vascular endothelial growth factor (VEGF) during folliculogenesis and corpus luteum formation in the human ovary.

Vascular endothelial growth factor (VEGF) has been suggested to be involved in angiogenesis and microvascular hyperpermeability. We examined immunohistochemically the expression of VEGF in the granulosa and theca cells, along with that of proliferating cell nuclear antigen (PCNA), in the vascular endothelium during the course of follicular development and corpora lutea formation in human ovaries. The immunolocalization of VEGF in these cells was compared with that of another putative angiogenic factor, basic fibroblast growth factor (bFGF). The granulosa cells in the primordial and primary follicles were VEGF negative, but at the preantral stage, the granulosa cells showed weakly positive immunostaining for VEGF. However, the VEGF immunostaining in the granulosa cells was weak throughout the folliculogenesis. In contrast, the theca interna cells of developing follicles showed strong staining for VEGF, which was well correlated with the PCNA positivity in the vascular endothelial cells in the thecal layer. In the atretic follicles, the granulosa and theca cells were VEGF negative. In the corpora lutea, VEGF was strongly expressed in both granulosa and theca lutein cells in the early luteal phase when the PCNA positivity in the endothelium increased, but the VEGF staining in these cells became weak in the mid- and late luteal phases. Accordingly, the PCNA positivity in the vascular endothelium was well correlated with the expression of VEGF in the theca cells during follicular development and atresia, and that in the granulosa and theca lutein cells in corpora lutea formation and regression. In addition, the immunolocalization of VEGF was different from that of bFGF.

Biopsy↗

PubMed1996

Fluorometric determination of phenol and p-cresol in urine by precolumn high-performance liquid chromatography using 4-(N-phthalimidinyl)benzenesulfonyl chloride.

A sensitive HPLC method for simultaneous determination of phenol and p-cresol in urine was developed. After acid hydrolysis of urine was conducted, phenol and p-cresol were extracted with diisopropyl ether and derivatized with 4-(N-phthalimidinyl)benzenesulfonyl chloride to give fluorescent sulfonyl esters. The labeling reactions were completed at 75 degrees C for 10 min. The fluorescent derivatives were separated on a reversed-phase column by a gradient elution with acetonitrile-water and detected by fluorescence measurement of excitation at 300 nm and emission at 410 nm. The detection limits (signal-to-noise ratio = 3) for phenol and p-cresol were 0.17 and 0.25 pmol per injection, respectively. The within-day and day-to-day relative standard deviations were 2.31-3.76 and 4.36%, respectively, for phenol and 1.99-4.56 and 3.71%, respectively, for p-cresol. The concentrations (means) of phenol and p-cresol in normal human urine were 67.3 and 167.9 nmol/mg creatinine, respectively.

Adult↗

PubMed1996

Definition of the mitogenic factor (MF) as a novel streptococcal superantigen that is different from streptococcal pyrogenic exotoxins A, B, and C.

Human T cell activation by recombinant mitogenic factor (rMF) was investigated in comparison with that by recombinant streptococcal pyrogenic exotoxins (rSPE) A, B, and C. Recombinant MF, rSPEA, and rSPEC were mitogenic for peripheral blood mononuclear cells (PBMC), whereas rSPEB was not. Recombinant MF required only HLA-DR for the stimulation of PBMC, as determined using monoclonal antibodies (mAb) to HLA class II molecules and the mouse L cells transfected with HLA class II molecules. Recombinant SPEA and rSPEC required HLA-DR or HLA-DQ molecule. Recombinant MF selectively stimulated V beta 2, V beta 7, V beta 8, V beta 18 and V beta 21-bearing T cells, whereas rSPEA and rSPEC activated V beta 2 and V beta 6-bearing T cells as evaluated by the quantitative T cell receptor (TCR) analytical method. No clonality was observed in the nucleotide sequences of complementarity determining region 3 of TCR V beta in T cells responding to rMF. The profiles of cytokine production by PBMC in response to rMF, rSPEA, and rSPEC were quite similar. In summary, these results demonstrate that both HLA class II molecules and the TCR V beta required for rMF-mediated T cell activation are distinct from those required for rSPEA or rSPEC-mediated activation. Therefore, the MF is a novel streptococcal super-antigen which is different from SPEA, SPEB, and SPEC.

Bacterial Proteins↗

PubMed1996

Spontaneous conception and intrauterine pregnancy in a symptomatic missed abortion of ectopic pregnancy conceived in the previous cycle.

We encountered a rare case of combined intrauterine and extrauterine pregnancy that occurred following separate spontaneous ovulations. A 33 year old woman visited our hospital with the chief complaint of abdominal pain on April 16, 1993. Her last menstruation was from March 23 for 6 days. However, the urinary human chorionic gonadotrophin (HCG) on April 19 was 1024 IU/l. Pelvic examination and ultrasonography indicated an extrauterine pregnancy, which was confirmed by laparotomy and histological identification of trophoblast cells. The urinary HCG concentration markedly decreased after the operation. However, the HCG level increased again on the fifth post-operative day, and a gestational sac (11 mm) was identified in the uterine cavity on the 11th post-operative day, indicating that this intrauterine pregnancy was established following spontaneous ovulation which occurred before the removal of the extrauterine pregnancy. This case indicates that a combined pregnancy can occur not only after simultaneous multiple ovulations but also after the separate spontaneous ovulations.

Adult↗

PubMed1996

Characterization of spirochetes isolated from ticks (Ixodes tanuki, Ixodes turdus, and Ixodes columnae) and comparison of the sequences with those of Borrelia burgdorferi sensu lato strains.

Ixodes persulcatus serves as a tick vector for Borrelia garinii and Borrelia afzelii in Japan; however, unidentified spirochetes have been isolated from other species of ticks. In this study, 13 isolates from ticks (6 from Ixodes tanuki, 6 from Ixodes turdus, and 1 from Ixodes columnae) and 3 isolates from voles (Clethrionomys rufocanus) were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, rRNA gene restriction fragment length polymorphism, partial sequencing of the outer surface protein C (OspC) gene, whole DNA-DNA hybridization, and 16S rRNA gene sequence comparison. All of the results revealed that these Borrelia strains clearly represent at least two new species. A third is also likely, although additional strains have to be isolated and characterized before a separate species is designated. We designated all isolates of I. tanuki and C. rufocanus as group Hk501 and all isolates of I. turdus as group Ya501. Phylogenetic analysis based on 16S rRNA gene sequences distinguished these Borrelia strains from those belonging to hitherto known Borrelia species. Furthermore, the genomic groups, each with its own tick vectors with enzootic cycles, were quite different from each other and also from those of Lyme disease Borrelia species known to occur in Japan. The results of 16S rRNA gene sequence comparison suggest that the strain Am501 from I. columnae is related to group Hk501, although its level of DNA relatedness is less than 70%.

Amino Acid Sequence↗

PubMed1996

Determination of total hydroxyproline and proline in human serum and urine by HPLC with fluorescence detection.

A method to measure total hydroxyproline (Hyp) and proline (Pro) in human serum and urine by HPLC was developed. Hyp and Pro in acid hydrolysates of serum and urine were derivatized with 4-(5,6-dimethoxy-2-phthalimidinyl)phenysulfonyl chloride after treatment with o-phthaladehyde and cleanup on Bond Elut C18 column. The derivatives of imino acids were separated on a reversed phase column by gradient elution with acetonitrile and phosphate buffer (1 mmol/l, pH 7) and detected by fluorescence measurement at 315 nm (excitation) and 385 nm (emission). Detection limits for both Hyp and Pro were 10 fmol per injection. The within-day and day-to-day relative standard deviations for Hyp and Pro in serum and urine were less than 3.19%. The recoveries of Hyp and Pro added to serum and urine were about 100%. The present method was applied to determine total Hyp and Pro in serum and urine from normal subjects and patients with chronic renal failure.

Adult↗

PubMed1995

Determination of free hydroxyproline and proline in human serum by high-performance liquid chromatography using 4-(5,6-dimethoxy-2-phthalimidinyl)phenylsulfonyl chloride as a pre-column fluorescent labelling reagent.

A fluorescent labelling reagent, 4-(5,6-dimethoxy-2-phthalimidinyl)phenylsulfonyl chloride, was designed for the determination of amines by precolumn HPLC and was applied to the simultaneous determination of hydroxyproline and proline in serum. The reagent reacted with hydroxyproline and proline at 30 degrees C for 10 min to produce the fluorescent derivatives, which were separated on a reversed-phase column by gradient elution with phosphate buffer (1 mmol l-1, pH 7) and acetonitrile and detected by fluorescence measurement at 315 nm (excitation) and 385 nm (emission). The detection limits (signal-to-noise ratio = 3) for both hydroxyproline and proline were 10 fmol per injection. The within-day (n = 10) and day-to-day (n = 5) relative standard deviations using human sera were less than 2.16% and 2.75%, respectively, for hydroxyproline and less than 2.30% and 3.25%, respectively, for proline. The concentrations of free hydroxyproline and proline in normal human sera (n = 13) were 5.6-18.0 and 137.6-252.6 mumol l-1, respectively. The proposed method was also applied to the determination of hydroxyproline and proline in sera from patients with chronic renal failure. The mean concentrations of hydroxyproline and proline in chronic renal failure were about 2.6 and 1.6 times higher, respectively, than those in normal human sera.

Chromatography, High Pressure Liquid↗

PubMed1995

Genetic and phenotypic analysis of Borrelia miyamotoi sp. nov., isolated from the ixodid tick Ixodes persulcatus, the vector for Lyme disease in Japan.

The ixodid tick Ixodes persulcatus is the most important vector of Lyme disease in Japan. Most spirochete isolates obtained from I. persulcatus ticks have been classified as Borrelia burgdorferi sensu lato because of their genetic, biological, and immunological characteristics. However, we found that a small number of isolates obtained from I. persulcatus contained a smaller 38-kDa endoflagellar protein and single 23S-5S rRNA gene unit. Representative isolate HT31T (T = type strain) had the same 23S rRNA gene physical map as Borrelia turicatae. The DNA base composition of strain HT31T was 28.6 mol% G+C. DNA-DNA hybridization experiments revealed that strain HT31T exhibited moderate levels of DNA relatedness (24 to 51%) with Borrelia hermsii, B. turicatae, Borrelia parkeri, and Borrelia coriaceae. However, the levels of DNA reassociation with the previously described Lyme disease borreliae (B. burgdorferi, Borrelia garinii, and Borrelia afzelii) were only 8 to 13%. None of the previously described species examined exhibited a high level of DNA relatedness with strain HT31T. In addition, the 16S rRNA gene sequence (length, 1,368 nucleotides) of strain HT31T was determined and aligned with the 16S rRNA sequences of other Borrelia species. Distance matrix analyses were performed, and a phylogenetic tree was constructed. The results showed that isolate HT31T is only distantly related to both previously described Lyme disease borreliae and relapsing fever borreliae. Thus, the spirochetes isolated from I. persulcatus and closely related isolates should be classified as members of a new Borrelia species. We propose the name Borrelia miyamotoi sp. nov. for this spirochete; strain HT31 is the type strain.

Animals↗