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T Ushijima

Publications and source records attributed to T Ushijima.

At least 91 records · Page 5Linked to original sources
PubMed1996

Microsatellite instability and loss of heterozygosity on chromosome 10 in rat mammary tumors induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine.

Microsatellite instability (MI) and loss of heterozygosity (LOH) were examined in mammary tumors induced in Sprague-Dawley x F344 F1 female rats by 2-amino-l-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Examination of 62 microsatellite loci revealed MI in nine of 15 (60%) PhIP-induced mammary tumors, and five of these MI-positive tumors had mutations in more than one microsatellite locus. In contrast, two of 12 (17%) 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumors were MI positive but had mutations at only one locus each. Further, by using 37 polymorphic markers specific LOH was observed in four of 15 PhIP induced mammary tumors on distal parts of rat chromosome 10, which is homologous to human chromosome 17q with no background level of LOH. Similarly, DMBA-induced mammary tumors showed specific LOH on the same region of chromosome 10. These data suggest that mismatch-repair deficiency and loss of chromosome 10 are involved in carcinogenesis of PhIP-induced rat mammary tumors.

Animals↗
PubMed1996

Internal thoracic artery graft function during exercise assessed by transthoracic Doppler echography.

BACKGROUND: Noninvasive quantitative assessment of internal thoracic artery (ITA) graft function at rest and during exercise is important in patients who have undergone coronary artery bypass grafting. METHODS: Blood flow in the ITA graft was measured using transthoracic color Doppler echography before and after operation in 50 patients who underwent coronary artery grafting using an ITA to the left anterior descending artery. The patients were divided into three groups according to the degree of coronary stenosis and previous anterior myocardial infarction: Group 1 included 12 patients with severe (90% or more) coronary stenosis accompanied by anterior infarction. Group 2 included 26 patients with severe coronary stenosis without anterior infarction. Group 3 included 12 patients with moderate (75% or less) coronary stenosis without anterior infarction. Transthoracic echographic images of the ITA were obtained through the first intercostal space using a 7.5-MHz probe, and the diameter and cross-sectional area of the ITA were measured on B-mode imaging. Systolic, diastolic, and mean blood flow velocity and volume were measured by the Doppler method. RESULTS: Internal thoracic artery diameter increased significantly from 2.2 mm to 2.4 mm after operation. The ITA flow patterns in both flow velocity and volume changed from systolic-dominant to diastolic-dominant after operation. Postoperative ITA graft flow was 82 +/- 24 mL/min, 53 +/- 30 mL/min, and 31 +/- 15 mL/min (p < 0.01, group 1 versus 3; p < 0.05, group 1 versus 2) and percent diastolic fraction of ITA flow was 72%, 68%, and 62% (not significant) in groups 1, 2, and 3, respectively. Compared with intraoperative ITA flow, which was measured using an ultrasound transit-time flowmeter, postoperative ITA graft flow was increased in group 1, but not changed in group 2 or 3. The ITA graft flow was measured before and after exercise in 19 patients and was compared with ITA flow in 10 normal control subjects. The ITA graft flow increased significantly with exercise in all patients in the three groups. Percent diastolic fraction of ITA flow increased significantly with exercise in patients with severe coronary stenosis (groups 1 and 2), but decreased significantly in patients with moderate stenosis (group 3). CONCLUSIONS: Changes in native coronary artery and ITA graft may be predicted by assessing ITA flow pattern during exercise. Transthoracic color Doppler echography is a clinically useful noninvasive method of assessing ITA graft function at rest and during exercise.

Adult↗
PubMed1996

Is internal thoracic artery grafting suitable for a moderately stenotic coronary artery?

Grafting an internal thoracic artery to a coronary artery with moderate stenosis remains controversial. Competitive flow from the native coronary artery has been proposed as the cause of distal narrowing and ultimate failure of the internal thoracic artery graft. We investigated intraoperative phasic blood flow in internal thoracic arteries grafted to coronary arteries with various degrees of stenosis and the influence of stenosis on postoperative angiographic findings. One hundred patients who underwent coronary artery bypass grafting of an internal thoracic artery to the left anterior descending coronary artery were divided into three groups according to degree of coronary stenosis. Group 1 included 39 patients who had 75% or less stenosis, group 2 included 34 patients with stenosis from 76% to 90%, and group 3 included 27 patients with stenosis greater than 90%. Mean flow and peak systolic flow of internal thoracic artery graft in group 1 were lower than those in group 2 (p < 0.01, p < 0.05). Peak diastolic flow in group 1 showed no difference from flows in groups 2 and 3. In eight patients in group 1, internal thoracic artery flow showed a predominant diastolic peak with characteristic systolic reversal as a result of competitive flow from the native coronary artery. Angiography at 1 month showed that the internal thoracic artery graft was patent in every case. Relative contributions of native coronary artery and internal thoracic artery flow to distal perfusion differed among the three groups (p < 0.001). In group 1, 15% of patients showed native-dominant flow, 62% showed balanced flow, and 23% showed internal thoracic artery-dependent flow. In group 2, 9% of patients showed native-dominant flow, 29% showed balanced flow, and 62% showed internal thoracic artery-dependent flow. In group 3, 96% of patients showed internal thoracic artery-dependent flow. String sign of the internal thoracic artery graft developed in only three patients; in two of these patients internal thoracic arteries were grafted to coronary arteries with stenosis of 50% or less and in the other patient there was competitive flow from a diagonal vein graft. Eleven of 13 internal thoracic arteries grafted to coronary arteries with stenosis of 50% or less did not show string sign. Competitive flow from a moderately stenotic coronary artery did not predispose the patient toward string sign of the internal thoracic artery graft in the presence of substantial diastolic internal thoracic artery flow. We conclude that internal thoracic artery grafting is acceptable for a moderately stenotic coronary artery.

Adult↗
PubMed1996

Rare activation of the human c-Ha-ras transgene of mice in hemangioendothelial sarcomas and liver tumors induced by Glu-P-1.

A transgenic mouse (Tg), having the human c-Ha-ras proto-oncogene, has been demonstrated to develop hemangioendothelial sarcomas (HESs) which are associated with the transgene mutation, but not to develop liver tumors. In this study, we examined the effects of 2-amino-6-methyldipyrido [1,2-a:3',2'-d] imidazole (Glu-P-1), a food-borne carcinogen, which has been demonstrated to induce HESs and liver tumors in CDF1 mice, on Tg mice. Chronic administration of 0.05% Glu-P-1 in the diet induced HESs in Tg (7/17), but not in 18 non-transgenic mice (N-Tg). With basal diet, two out of 17 Tg but none of 22 N-Tg, developed HESs. In contrast, Glu-P-1 administration induced liver tumors, both in Tg and in N-Tg; 16/17 in Tg and 13/18 in N-Tg. The incidence of hepatocellular carcinomas in Tg was higher than that in N-Tg (8/17 versus 3/18). With basal diet, only one out of 17 Tg and none of 22 N-Tg developed liver tumors. The Ha-ras mutation in tumors developed by the groups administered Glu-P-1, was examined. No mutations were detected in the transgene and mouse c-Ha-ras genes in all three HESs examined. In contrast, when 29 liver tumors taken from Tg were examined, two mutations of the transgene were detected in two HCCs. No mouse c-Ha-ras gene mutations were detected in any of the 47 liver tumors examined, which had developed in Tg and N-Tg mice. These results suggest that the transgene plays a role in the development of HESs induced by Glu-P-1, but not as a result of its mutation. Further, the transgene plays no significant role in the development of liver tumors induced by Glu-P-1, but does play a role in the malignant conversion of some liver tumors, as a result of its mutation.

Animals↗
PubMed1996

Identification of 29 rat genetic markers by arbitrarily primed polymerase chain reaction.

The number of genetic markers for the rat is still limited, in spite of its wide use in cancer research. To facilitate accurate mapping of both established and novel rat genetic markers, we constructed a linkage map by genotyping 105 F2 rats from ACI/N (ACI) and BUF/Nac (BUF) crosses. This map consists of 120 genetic markers that had been previously reported, mainly by two research groups, but had not been integrated. To find new genetic markers, the arbitrarily primed polymerase chain reaction (AP-PCR) was applied to detect polymorphic bands between ACI and BUF rats. After testing 56 single primers and 12 combinations of primers, we found 36 bands produced by 16 single primers and two combinations to be reliably polymorphic between ACI and BUF rats. The 36 bands were typed in the 105 F2 rats, and 29 of them could be linkage-mapped. AP-PCR is thus useful to detect new genetic markers in laboratory strains of rats.

Animals↗
PubMed1996

Human exposure to carcinogenic heterocyclic amines and their mutational fingerprints in experimental animals.

Heterocyclic amines (HCAs) are mutagens/carcinogens to which humans are exposed on almost a daily basis. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhlP) is the most abundant of the various carcinogenic HCAs (present at a level of 0.56 to 69.2 ng/g of cooked meat or fish), with 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MelQx) following it at 0.64 to 6.44 ng/g. HCAs have been found in the urine of healthy people who consume ordinary diets, while patients receiving parenteral alimentation lack, for example, PhlP and MelQx in their urine. Based on the concentrations of PhlP and MelQx in urine samples from 10 healthy volunteers, daily intake of MelQx in Japanese was calculated to be 0.3 to 3.9 micrograms/person, while that of PhlP was 0.005 to 0 micrograms. The Japanese consume more MelQx than Americans, whereas Japanese intake of PhlP was about one-third that of Americans. MelQx-DNA adducts have also detected in Japanese Kidney, colon, and rectum samples using the 32P-postlabeling method followed by identification using high-performance liquid chromatography (HPLC) analysis; the levels were 0.18, 1.8, and 1.4 per 10(9) nucleotides, respectively. In addition, we elucidated the mutational fingerprints of Phlp by analyzing Apc mutations in rat colon cancers induced by this carcinogen. Four of eight tumors had a total of five mutations in the Apc gene, four of which featured a guanine deletion from 5'-GTGGGAT-3' sequences. This specific mutation spectrum may be used as a fingerprint of PhlP in evaluating its risk potential for human colon carcinogenesis. Mutations were not found in similar 2-amino-3-methylimidazo[4,5-f]quinoline-induced colon lesions. Microsatellite instability was detected in both colon and mammary tumors induced by PhlP. The mechanisms involved in this development of microsatellite instability in PhlP. The mechanisms involved in this development of microsatellite instability in PhlP-induced cancers remain to be elucidated.

Amines↗
PubMed1995

2-(5-hydrazinocarbonyl-2-oxazolyl)-5,6-dimethoxybenzothiazole as a precolumn fluorescence derivatization reagent for carboxylic acids in high-performance liquid chromatography and its application to the assay of fatty acids in human serum.

A unique acid hydrazide, 2-(5-hydrazinocarbonyl-2-oxazolyl)-5,6-dimethoxybenzothia zole, that is characterized by the benzothiazole structure conjugated to an oxazoline moiety was synthesized, and its applicability as a precolumn derivatization reagent for carboxylic acids in HPLC was examined in view of sensitivity and separability. The sensitivity of the hydrazide for carboxylic acids was determined using lauric acid to be 0.1 pmol (per 10-microliters injection volume) at a signal-to-noise ratio of 3. The reagent allowed rapid assays of carboxylic acids within 20 min with satisfactory separability. The method was applied to the determination of fatty acids in human sera from healthy volunteers as well as from patients with diabetes or thyroid dysfunction.

Carboxylic Acids↗
PubMed1995

Germline mutation of BRCA1 in Japanese breast cancer families.

We analyzed germline mutations of the BRCA1 gene in 18 Japanese breast cancer families and two Japanese breast-ovarian cancer families. In two site-specific breast cancer families, the same mutation was detected; a nonsense mutation at codon 63 encoding a truncated small protein. It was demonstrated that the mutant allele cosegregated with breast cancer patients within a family and was absent in healthy Japanese, suggesting a breast cancer-predisposing allele. The average age at diagnosis was 44 and 55 years in each family with BRCA1 mutation. No bilateral breast cancer patients were present in the BRCA1 mutation-positive families, although five were present in the BRCA1-negative families. No germline mutations of BRCA1 were detected in the two breast-ovarian cancer families examined in this study, although BRCA1 mutation plays a major role in breast-ovarian cancer families in Western countries. Thus, the proportion of families who inherit the mutated BRCA1 allele seems to be small among Japanese breast cancer families and Japanese breast-ovarian cancer families.

Adult↗
PubMed1995

Specific 5'-GGGA-3'-->5'-GGA-3' mutation of the Apc gene in rat colon tumors induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine.

The APC gene plays a major role in human colon carcinogenesis. We determined the genomic structure of the rat Apc gene, and we analyzed mutations in colon tumors induced in F344 rats by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), potent carcinogens contained in ordinary daily human food. Of eight PhIP-induced tumors, one tumor had two Apc mutations, two tumors had a mutation with loss of the normal allele, and one had a mutation. Two of the above five mutations were at nucleotide 1903, one at 2605, and two at 4237, all being a deletion of a guanine base at the 5'-GGGA-3' site and resulting in truncation of the APC protein. Of 13 IQ-induced tumors, 2 had an Apc mutation with loss of the normal allele. The two mutations were a missense mutation (T-->C) at nucleotide 1567 and a nonsense mutation (C-->T) at 2761. Alteration of the Apc gene was shown to play a more important role in PhIP-induced than in IQ-induced rat colon carcinogenesis. PhIP-induced tumors are characterized by their specific and unique mutation, which may be useful for mutational fingerprinting of human cancers.

Animals↗
PubMed1995

Mutation, loss of heterozygosity, and recombination of the p53 gene in mouse forestomach tumors induced by 2-amino-3,4-dimethylimidazo[4,5-f]quinoline.

2-Amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), a food mutagen, induces forestomach tumors in CDF1 mice. We established a polymerase chain reaction (PCR)-single-strand conformation polymorphism (SSCP) analysis system to detect mutations in the mouse p53 gene exons 2-10, which encompass all five regions conserved among species, and a system to examine loss of heterozygosity (LOH) that uses newly identified polymorphisms between BALB/c and DBA mice, the parental strains of CDF1 mice. Four original forestomach tumors (one papilloma, two carcinomas, and one lymph-node metastasis) and four cell lines derived from four independent forestomach tumors were examined with the PCR-SSCP system and by polymorphism analysis. Of the four original tumors, the papilloma had a G-->A transition at the second position of codon 171, and one carcinoma had a G-->T transversion at the second position of codon 113 with loss of the wild-type allele, whereas the other two carcinomas had no detectable mutations. Of the four cell lines, two had a base substitution and LOH, and the other two had double mutations (a base substitution and a deletion). By amplification of the double mutations in a fragment, the two cell lines were shown to have four kinds of alleles, indicating induction of recombination within the p53 gene. Our results show that our PCR-SSCP analysis system is efficient for detecting p53 mutations in mouse genomic DNA and that alteration of the p53 gene plays a significant role in MeIQ-induced mouse forestomach carcinogenesis.

Animals↗
PubMed1995

No involvement of Ki-ras or p53 gene mutations in colitis-associated rat colon tumors induced by 1-hydroxyanthraquinone and methylazoxymethanol acetate.

1-Hydroxyanthraquinone (1-HA), which is present in some herbs, and methylazoxymethanol (MAM) acetate, a metabolite of azoxymethane, show synergistic carcinogenicity in rat colon, and 1-HA induces ulcerative changes with simultaneous severe inflammation of the entire colon. In this study, mutations in Ki-ras (exons 1 and 2) and p53 (exons 4-7) were studied by polymerase chain reaction (PCR)-single-strand conformation polymorphism (SSCP) analysis. Of 18 adenomas and 38 adenocarcinomas induced in male F344 rats (52 tumors induced by 1-HA plus MAM acetate, three by 1-HA alone, and one by MAM acetate alone), no mutations in Ki-ras or p53 were detected under two conditions of PCR-SSCP analysis. Because human colon carcinomas from patients with ulcerative colitis have a very low incidence of Ki-ras mutation, this experimental system would be a good animal model of human colon carcinomas with ulcerative colitis and of human colon carcinomas without Ki-ras or p53 mutations.

Animals↗
PubMed1995

Construction of a phylogenetic tree for inbred strains of rat by arbitrarily primed polymerase chain reaction (AP-PCR).

We constructed a tentative genealogic tree of 13 inbred rat strains using genetic markers identified by the AP-PCR (Arbitrarily Primed Polymerase Chain Reaction) technique, which consists of PCR amplification under low stringency conditions, with only one oligonucleotide as both forward and reverse primers. We obtained discrete-state genotypes of 264 loci, presumably covering the whole rat genome. Computational analysis of this panel allowed the construction of a genealogic tree representing the relations among the 13 strains. The accordance of the present results with available data on the histories of some of these strains validates the use of the AP-PCR technique for studies on phylogeny.

Animals↗
PubMed1995

A rapid method for detection of mutations in the lacI gene using PCR-single strand conformation polymorphism analysis: demonstration of its high sensitivity.

The lacI gene has been used as a target gene in various mutation assays. We modified single strand conformation polymorphism (SSCP) analysis by introducing restriction digestion to detect mutations in the gene rapidly, and determined the sensitivity of the method. The entire coding sequence and partial promoter region of the lacI gene were amplified by the polymerase chain reaction with [alpha-32P]dCTP in a 1247 base pair fragment, digested into eight restriction fragments, and analyzed by SSCP. The sensitivity of the method was assessed using 160 phages with lacI mutations, which were selected by assay of expression of beta-galactosidase after their infection into E. coli. Of the 160 mutants, 146 (91.3%) showed shifted bands in the first condition of SSCP analysis (without glycerol, 20 degrees C). The remaining 14 mutants were analyzed in a second condition (with 5% glycerol, 20 degrees C), and eight of them showed shifted bands (cumulatively 96.3% of the 160 mutants). The remaining six mutants were analyzed in a third condition (with 5% glycerol, 10 degrees C), and all of them showed shifted bands (cumulatively 100%). Sequencing of the restriction fragments with mobility shifts in the 160 mutants revealed 108 kinds of mutations, 100 (92.6%) being detected in the first condition, seven (cumulatively 99.1%) in the second condition, and one (cumulatively 100%) in the third condition. This method greatly reduced the time to identify lacI mutations, and allowed the detection of multiple mutations in one lacI mutant. The results also show that in general PCR-SSCP analysis is very sensitive when test fragments are shorter than about 250 base pairs and electrophoresis is performed under at least two conditions.

Animals↗
PubMed1995

Splicing mutations in the COL4A5 gene in Alport's syndrome: different mRNA expression between leukocytes and fibroblasts.

The COL4A5 gene from 40 patients with Alport's syndrome was examined using single-strand conformation substitution at the acceptor site (-2) of intron 50 and a G-to-C substitution at the donor site (+1) of intron 47, respectively. The transcript in peripheral leukocytes from the former had a 10-nucleotide deletion. This shortened transcript was derived from abnormal splicing in a cryptic acceptor site within exon 51. This could be translated into a protein with an alteration of three amino acids followed by premature termination, which eliminated 23 amino acids from the carboxyl end. Gene tracking revealed that the mother and a brother carried the mutant allele. In the latter, the transcript in leukocytes was normal, but that in cultured skin fibroblasts showed skipping of exon 47, the result being that 71 amino acids were absent. Glomerular basement membrane from the patient did not react with the anti-alpha 5(IV) antibody. His maternal grandmother, mother, and a sister, all with abnormal urinalysis, carried the mutant allele. Thus, the appearance of exons of the COL4A5 gene in leukocytes may differ from that in fibroblasts. If kidney mRNA is not available, mRNAs from cultured skin fibroblasts, in addition to leukocytes, can be used for gene analysis in subjects with Alport's syndrome.

Adolescent↗
PubMed1995

Genetic polymorphisms and susceptibility to cancer development.

Humans show heterogeneous susceptibility to cancer development, suggesting the involvement of various genetic backgrounds in control of the production of endogenous carcinogens, the metabolism of carcinogens, the repair of DNA damage, cell proliferation and defence mechanisms including immune reactions. Gastric cancer is the major cancer in Japan. However, little is known about the genes linked with its development. In 1967, we found that N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced gastric cancers in Wistar rats. Subsequently the Buffalo strain of rats was reported to be resistant to MNNG stomach carcinogenesis, while ACI rats were very sensitive. In a carcinogenesis study using F1 and F2 rats, we suggested that this trait of MNNG stomach carcinogenesis-resistance was regulated by a single autosomal dominant allele. The O6-methylguanine adduct levels in gastric mucosa induced by MNNG were the same in Buffalo and ACI rats, but cell proliferation induced by MNNG was much higher in ACI than Buffalo animals. Chromosome mapping of the gene responsible for susceptibility to MNNG-induced carcinogenesis is now in progress and its identification will hopefully give us clues to the involvement of genetic traits in susceptibility to gastric cancer in humans. In addition, the genetic background of susceptibility to breast cancer is also being studied. In Japan, about 5% of all cases of breast cancer are familial. We have studied BRCA1, the breast cancer susceptibility gene, as a determinant of susceptibility to breast cancer by linkage analyses in 11 families, but our results indicate that BRCA1 may not be important for development of familial breast cancer in Japanese.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗