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Biomedical subjects

T Yamagami

Publications and source records attributed to T Yamagami.

At least 91 records · Page 5Linked to original sources

Complete amino acid sequence of chitinase-a from bulbs of gladiolus (Gladiolus gandavensis).

The complete amino acid sequence of gladiolus bulb chitinase-a (GBC-a) was determined. First the tryptic peptides from GBC-a after it was reduced and S-carboxymethylated were sequenced and then the peptides were further studied by chemical cleavage of the enzyme. GBC-a consisted of 274 amino acid residues and had a molecular mass of 30,714 Da. Two consensus sequences essential for chitinase activity by plant class III chitinases were conserved in GBC-a, although its sequence similarity with plant class III chitinases was less than 20%. Sequence comparison of GBC-a with sequences of other proteins in a protein identification resource (PIR) showed that the GBC-a sequence was 33% similar to that of narbonin, a seed storage 2S globulin from narbon beans.

Amino Acid Sequence↗

Isolation and characterization of chitinase isoforms from the bulbs of four species of the genus Tulipa.

Six chitinase isoforms, designated TBC-1 to TBC-6, were purified to homogenity from the bulbs of four species (Tulipa bakeri, T. tarda, T. turkestanica, and T. praestans) of the genus Tulipa by CM-cellulose column chromatography, Butyl-Toyopearl 650M hydrophobic column chromatography, gel filtration on Sephadex G-75, and Mono-S fast protein liquid chromatography (FPLC). The chitinases had molecular weights of 30,000 and isoelectric points of 5.2 to 6.1. These chitinases were found to proteins with similar amino acid compositions and N-terminal sequences. The tulip chitinases all had two half-cystine residues, one more than gladiolus bulb class IIIb chitinase, but many fewer than chitinases of plant class I (15-17 Cys residues/mol), II (5-8 Cys residues/mol), or III (6 Cys residues/mol). The N-terminal sequences of tulip chitinases were similar to the sequence of the gladiolus chitinase, but did not resemble sequence of any class of plant chitinase. The optimal pH of these chitinases toward glycolchitin was pH 5. TBC-1 hydrolyzed (GlcNAc)6 into (GlcNAc)2, (GlcNAc)3, and (GlcNAc)4, and hydrolyzed (GlcNAc)5 into (GlcNAc)2 and (GlcNAc)3.

Chitin↗

Complete amino acid sequence of chitinase-A from leaves of pokeweed (Phytolacca americana).

The complete amino acid sequence of pokeweed leaf chitinase-A was determined. First all 11 tryptic peptides from the reduced and S-carboxymethylated form of the enzyme were sequenced. Then the same form of the enzyme was cleaved with cyanogen bromide, giving three fragments. The fragments were digested with chymotrypsin or Staphylococcus aureus V8 protease. Last, the 11 tryptic peptides were put in order. Of seven cysteine residues, six were linked by disulfide bonds (between Cys25 and Cys74, Cys89 and Cys98, and Cys195 and Cys208); Cys176 was free. The enzyme consisted of 208 amino acid residues and had a molecular weight of 22,391. It consisted of only one polypeptide chain without a chitin-binding domain. The length of the chain was almost the same as that of the catalytic domains of class IL chitinases. These findings suggested that this enzyme is a new kind of class IIL chitinase, although its sequence resembles that of catalytic domains of class IL chitinases more than that of the class IIL chitinases reported so far. Discussion on the involvement of specific tryptophan residue in the active site of PLC-A is also given based on the sequence similarity with rye seed chitinase-c.

Amino Acid Sequence↗

[Studies on carcinogenicity of asbestos with special reference to interaction between viruses and cultured cells].

To investigate effects of asbestos on the process of cancer development, the capacity of asbestos which increases cellular uptake of external carcinogens was tested for asbestos-mediated viral RNA transfection in cultured cells. For the transfection, crocidolite, amosite, anthophyllite and chrysotile were placed onto Vero-E6 cells with poliovirus RNA inoculum, respectively. All asbestos samples mediated viral RNA transfection compared with the background including only viral RNA. The transfection was much greater with combined exposure to asbestos and kaolin than with exposure to asbestos alone.

Animals↗

High incidence of chemotherapy-induced acral erythema in female patients with non-Hodgkin's lymphoma treated with the VACOP-B regimen.

Seven patients, all females out of 29 with non-Hodgkin's lymphoma (NHL) (16 males and 13 females) treated with the VACOP-B regimen utilizing granulocyte-colony-stimulating factor (G-CSF) support developed chemotherapy-induced acral erythema (CAE). In contrast, none of 32 patients with NHL who were treated with CHOP, MACOP-B, or biweekly CHOP regimens without G-CSF developed CAE. Total dose intensities of VACOP-B regimen were higher than those of the three other regimens. However, no significant difference in dose intensities of each drug in the patients treated with the VACOP-B regimen was found between male and female patients and between female patients with or without CAE. The cause of the high incidence of CAE (7/13) in the female patients treated with VACOP-B regimen remains unknown. However, female sex hormones may increase susceptibility to CAE. Since the occurrence of CAE interrupts intensive chemotherapy and reduces the cure rate, high risk patients for CAE should be carefully monitored for early symptoms and signs of CAE and should be treated early and appropriately.

Antineoplastic Combined Chemotherapy Protocols↗

Postlaminectomy adhesion of the cauda equina. Changes of postoperative vascular permeability of the equina in rats.

STUDY DESIGN: The authors evaluated the vascular permeability changes of the cauda equina after lumbar laminectomy in rats. OBJECTIVES: To clarify the early vascular responses in postlaminectomy adhesive arachnoiditis. SUMMARY OF BACKGROUND DATA: Laminectomy-induced cauda equina adhesion has been visualized by postoperative serial magnetic resonance imaging in humans. In laminectomized rats, fibrinous exudation was apparent among the adhered cauda equina. To date, there has been no report of substantial changes in vascular permeability in the cauda equina after lumbar laminectomy. METHODS: Wistar rats laminectomized from L5-L6 were used for four studies; 1) Evans blue albumin tracer study; 2) horseradish peroxidase tracer study; 3) lanthanum tracer study; and 4) fluoroscein isothiocyanate-dextran (molecular weights: 20K, 70K, and 150K) tracer study. Untreated rats were used as controls. RESULTS: 1) In the laminectomized (L5-L6) area, the cauda equina tended to conglomerate from 3 hours after surgery; this was relatively resolved by 6 weeks. Extravascular leakage of Evans blue albumin in the cauda equina appeared at 3 hours after surgery and reached a plateau at 24 hours; it mostly disappeared by 6 weeks. At 24 hours after surgery, the extent of Evans blue albumin extravasation was seen in one or more levels adjacent to the laminectomized area. Evans blue albumin leakage and cauda equina adhesion started to appear in nearly a same phase after laminectomy, followed by a plateau of vascular permeability facilitating complete cauda equina adhesion by 24 hours after surgery. The restoration of Evans blue albumin leakage was noted 1 week after surgery, and tended to precede recovery of the cauda equina adhesion; 2) in the laminectomy group, Horseradish peroxidase penetrated from the lumen to the extracellular space beyond the basal laminae of the endothelial cells. Numerous horseradish peroxidase-labeled vesicles in the endothelial cells were found; 3) There was a significant increase in lanthanum distribution in the endothelial cytoplasm at the laminectomized levels. Lanthanum filled the interendothelial clefts beyond the tight junction, but did not penetrate through the perivascular basal laminae; 4) Fluoroscein isothiocyanate-dextran (20K, 70K, 150K) permeability increased in the laminectomy group, but there was no difference in extravasation of fluoroscein isothiocyanate-dextran, irrespective of the molecular weight. CONCLUSION: Laminectomy consistently induced an increase in vascular permeability in the cauda equina, an increase of vesicular transport in the endothelial cell, and opening of the tight junction early after laminectomy, suggesting breakdown of the blood nerve barrier in the cauda equina. The accelerated permeability may enhance cauda equina adhesion.

Animals↗

A case of internal carotid artery occlusion complicating embolization of the external carotid artery system.

A case of visual disturbance by embolization through the external carotid artery (ECA) after internal carotid artery (ICA) occlusion is reported. A 65-year-old female presented left ICA occlusion 14 years ago. She was admitted with acute deterioration of left visual acuity. Ophthalmologically, the retinal central artery was occluded. Cerebral angiography disclosed an irregular wall at the left ICA stump and stenosis of the maxillary artery. Carotid endarterectomy of the common carotid artery, ECA and residual ICA was performed and atheromatous plaque with ulceration was removed. The postoperative course was uneventful. We discuss cases of ICA occlusion in relation to visual impairment.

Aged↗

Myoepithelial hamartoma of the ileum causing intussusception in an infant

Intussusception is a relatively common pediatric gastrointestinal emergency, although the etiology in most cases remains unknown. In a small number of cases the lead point is a tumor or tumor-like lesion of the small bowel. We report an unusual case of a myoepithelial hamartoma of the ileum causing an intussusception in a 4-month-old boy. This is the second such report in the literature.

Journal Article↗

Aberrant overexpression of the Wilms tumor gene (WT1) in human leukemia.

To clarify whether the expression of the WT1 gene in leukemic cells is aberrant or merely reflects that in normal counterparts, the expression levels of the WT1 gene were quantitated for normal hematopoietic progenitor cells. Bone marrow (BM) and umbilical cord blood (CB) cells were fluorescence-activated cell sorting (FACS)-sorted into CD34+ and CD34- cell populations, and the CD34+ cells into nine subsets (CD34+ CD33-, CD34+ CD33+, CD34+ CD38-, CD34+ CD38+, CD34+ HLA-DR-, CD34+ HLA-DR+, CD34+ c-kit(high), CD34+ c-kit(low), and CD34+ c-kit-) according to the expression levels of CD34, CD33, CD38, HLA-DR, and c-kit. Moreover, acute myeloid leukemic cells were also FACS-sorted into four populations (CD34+ CD33-, CD34+ CD33+, CD34- CD33+, and CD34- CD33-). FACS-sorted normal hematopoietic progenitor and leukemic cells and FACS-unsorted leukemic cells were examined for the WT1 expression by quantitative reverse transcriptase-polymerase chain reaction. The WT1 expression in the CD34+ and CD34- cell populations and in the nine CD34+ subsets of BM and CB was at either very low (1.0 to 2.4 x 10(-2)) or undetectable (< 10(-2)) levels (the WT1 expression level of K562 cells was defined as 1.0), whereas the average levels of WT1 expression in FACS-sorted and -unsorted leukemic cells were 2.4 to 9.3 x 10(-1). Thus, the WT1 expression levels in normal hematopoietic progenitor cells were at least 10 times less than those in leukemic cells. Therefore, we could not find any normal counterparts of BM or CB that expressed the WT1 at levels comparable with those in leukemic cells. These results indicate an aberrant overexpression of the WT1 gene in leukemic cells and imply the involvement of this gene in human leukemogenesis.

Acute Disease↗

Lineage- and differentiation stage-specific expression of LSM-1 (LPAP), a possible substrate for CD45, in human hematopoietic cells.

CD45, a transmembrane tyrosine phosphatase, is found on almost all nucleated hematopoietic cells and plays a crucial role in lymphocyte activation and differentiation. We recently achieved isolation of the human LSM-1 (hLSM-1) gene, whose product is a possible substrate for CD45, and we raised antibodies against the hLSM-1 protein. hLSM-1 expression in hematopoietic cells was examined with Northern and Western blot, fluorescence-activated cell sorter, and immunocytochemical analyses. It was found that in the lymphoid lineage, T and B lymphocytes as well as NK cells expressed LSM-1, whereas terminally differentiated plasma cells did not. As for the myeloid lineage, immature myeloid cells expressed LSM-1, whereas terminally differentiated granulocytes and monocytes did not. In the erythroid lineage, normal erythroblasts expressed very low levels of LSM-1, while erythroid cell lines (K562 and HEL) did not. Megakaryocytes did not express LSM-1. Both CD34+/CD33- and CD34+/CD33+ hematopoietic progenitor cells weakly expressed LSM-1. These results showed that LSM-1 is expressed in a lineage- and differentiation stage-specific fashion.

Blotting, Western↗

Myoepithelial hamartoma of the ileum causing intussusception in an infant.

Intussusception is a relatively common pediatric gastrointestinal emergency, although the etiology in most cases remains unknown. In a small number of cases the lead point is a tumor or tumor-like lesion of the small bowel. We report an unusual case of a myoepithelial hamartoma of the ileum causing an intussusception in a 4-month-old boy. This is the second such report in the literature.

Diagnosis, Differential↗

Primary obsessional slowness: long-term findings.

Cases of slowness among patients who spent large amounts of time to perform daily activities were first reported in 1974, and described as primary obsessional slowness (POS). It was observed that it was neither obsessive thoughts nor compulsions that directly hindered their daily activities. However, in more than 20 years following the original report, the diagnostic independence of POS remains controversial, some insisting that obsessional slowness can be explained as secondary. The authors experienced four cases in Japan which share the same characteristics as the original cases. Long-term observation and treatment has led us to support the diagnostic independence of POS. Slowness remained after other accompanying symptoms of obsessive-compulsive disorder had been successfully treated, showing that the slowness of our patients was not secondary.

Activities of Daily Living↗

Purification and characterization of two chitinase isoforms from the bulbs of gladiolus (Gladiolus gandavensis).

Two chitinase isoforms, designated GBC-a and GBC-b, were purified from the bulbs of gladiolus (Gladiolus gandavensis) using CM-cellulose column chromatography followed by Butyl-Toyopearl 650 M hydrophobic column chromatography, gel filtration on Sephadex G-75, and Mono-S FPLC. GBC-a and GBC-b are weakly acidic and weakly basic proteins with molecular masses of 30 kDa, and isoelectric points of 6.0 and 7.5, respectively. GBC-a and GBC-b were found to be homologous proteins with similar amino acid compositions and N-terminal sequences. The number of half-cystine residues in GBC-a and GBC-b was only one each, which is much lower than those of plant class I (15-17 Cys residues/mol), class II (5-8 Cys residues/mol), and class III (6 Cys residues/mol) chitinases. The N-terminal sequences of GBC-a and GBC-b were completely different from those of plant three classes of chitinases. The optimal pHs of these chitinases toward glycolchitin were pH 5. GBC-a hydrolyzed (GlcNAc)5 into (GlcNAc)2, (GlcNAc)3 and (GlcNAc)4, and (GlcNAc)5 into (GlcNAc)2 and (GlcNAc)3.

Amino Acid Sequence↗

Ischaemic heart disease deaths in a Japanese rural area evaluated by clinical records.

A community-based survey to examine the actual number of ischaemic heart disease (IHD) deaths was undertaken in the western area of Toyama, a rural area with a population of 209,000. IHD deaths (International Classification of Diseases [ICD], ninth revision, codes 410-414) and heart failure (HF) deaths (ICD code 428) according to the death certificate aged 15 to 74 in 1987 to 1990 were reevaluated from medical records using the criteria of the multinational monitoring of trends and determinants in cardiovascular disease (MONICA). Of 97 subjects with IHD and of 170 subjects with HF according to the death certificate, 85 cases (87.6%) and 143 cases (84.1%) could be examined, respectively. Using the MONICA criteria, of 85 subjects with IHD, 18 (21.2%) were reevaluated as "definite acute myocardial infarction (AMI)" and 30 (35.3%) as "possible AMI". On the other hand, of 143 subjects with HF, 1 (0.7%) were reevaluated as "definite AMI" and 12 (8.4%) as "possible AMI". From these results, the number of deaths as reevaluated IHD was estimated 104.5 in the study period, and it was 7.7% increase at least compared with the number of deaths judged from the death certificate.

Adolescent↗

[Enhancement pattern of hepatic metastases from colorectal cancer on CT arteriography].

The enhancement pattern of hepatic metastases from colorectal cancer on CT arteriography (CTA) was evaluated in 79 cases with 405 lesions. Sixty-two percent of overall lesions irrespective of size showed ring-like enhancement, and 33% of lesions less than 1 cm in diameter showed homogeneous enhancement. In pathological examination of 59 resected lesions, the enhancement pattern on CTA had a good correlation with the extent of tumor cellularity, necrosis or fibrosis in the lesions. And in 8.5% of resected lesions, the extent of enhancement on CTA was larger than that of perfusion defect on CT arterial portography, which was almost equal to the size in the resected specimen. In such lesions, the enhancement on CTA might include enhancement of normal hepatic parenchyma around the lesion.

Adenocarcinoma↗

Long-term follow-up of minimal residual disease in leukemia patients by monitoring WT1 (Wilms tumor gene) expression levels.

Thirty-one patients (27 with acute myeloid leukemia [AML], 2 with acute lymphocytic leukemia [ALL], and 2 with acute mixed lineage leukemia [AMLL]) treated with conventional chemotherapy (CHT) and 23 patients (13 AML, 5 ALL, and 5 with chronic myeloid leukemia [CML]) treated with allogeneic bone marrow transplantation (BMT) were monitored for WT1 expression levels in BM and peripheral blood (PB) by reverse transcriptase-polymerase chain reaction over a long-term period (mean, 29 months for CHT and 24 months for BMT). Sixteen of the patients in the CHT group and 3 in the BMT group who had achieved complete remission suffered clinical relapse. In 10 of these patients, WT1 expression that had returned to normal BM levels (< 10(-3); the WT1 expression level of K562 cells was defined as 1.0) after complete remission (CR) either gradually or rapidly increased again to abnormal levels 1 to 18 months (mean, 7 months) before clinical relapse became apparent. In another 9 patients, WT1 expression never returned to normal BM levels even after CR and the subsequent relapse was accompanied by a rapid increase in WT1 expression to levels higher than 10(-2) (10(-3) levels in PB). On the other hand, the remaining 35 patients (15 CHT and 20 BMT) maintained their CR. In 29 of these patients (11 CHT and 18 BMT), WT1 expression either gradually or rapidly decreased to normal BM levels, whereas in the other 6 (4 CHT and 2 BMT), low or very low levels of WT1 mRNAs (10(-3) to 10(-2) in BM and 10(-5) to 10(-3) in PB) remain detectable, but without any clinical signs of relapse. A clear correlation was found to exist between the minimal residual disease (MRD) detected in the paired BM and PB samples for all types of leukemias (AML, ALL, and CML), with MRD in PB being approximately one-tenth of that in BM. WT1 quantitation of 168 paired BM and PB samples showed that PB samples were superior to BM samples for the detection of MRD. We conclude that monitoring of WT1 expression levels in BM and PB makes it possible to rapidly assess the effectiveness of individual treatment and diagnose clinical relapse in the early stage for all leukemia patients regardless of the presence or absence of tumor-specific DNA markers.

Bone Marrow↗

Growth inhibition of human leukemic cells by WT1 (Wilms tumor gene) antisense oligodeoxynucleotides: implications for the involvement of WT1 in leukemogenesis.

We have previously reported expression of WT1 in acute leukemia. To elucidate its biological significance, we examined the effect of the suppression of the WT1 expression by WT1 antisense oligomers on the growth of the leukemic cells expressing WT1. When 20 different WT1 antisense (AS) oligomers covering from the 5' cap sites of the WT1 gene to the 3' end were examined for the inhibitory effect on the growth of K562 cells expressing WT1, four WT1 AS oligomers inhibited the cell growth, whereas WT1 sense and random sequence oligomers had no effect on the cell growth of K562. Moreover, WT1 AS oligomers significantly inhibited the growth of the clonogenic cells of fresh leukemic cells in six of 14 patients with acute myeloid leukemia, in one of two patients with chronic myelogenous leukemia (CML) chronic phase, and in one of one patient with CML blastic crisis. However, these oligomers did not inhibit normal colony-forming unit-granulocyte-macrophage. Western blot analysis clearly demonstrated the significant reduction in the WT1 protein levels in the K562 and fresh leukemic cells that were treated with the WT1 AS oligomers, confirming that the inhibitory effect of the WT1 AS oligomers on the cell growth operates via the reduction in the WT1 protein levels. These results show that WT1 plays an important role in leukemogenesis.

Base Sequence↗