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Biomedical subjects

T Gotoh

Publications and source records attributed to T Gotoh.

At least 91 records · Page 5Linked to original sources

Long-term effects of dietary alpha-linolenic acid from perilla oil on serum fatty acids composition and on the risk factors of coronary heart disease in Japanese elderly subjects.

Although important roles of dietary n-3 fatty acids in the prevention of coronary heart disease (CHD) have been suggested, long-term effects of dietary alpha-linolenic acid (ALA, 18:3n-3) have not yet been established under controlled conditions. We tested whether a moderate increase of dietary ALA affects fatty acids composition in serum and the risk factors of CHD. Oxidized LDL (OxLDL) was directly measured by ELISA using antibody specific to OxLDL. By merely replacing soybean cooking oil (SO) with perilla oil (PO) (i.e., increasing 3 g/d of ALA), the n-6/n-3 ratio in the diet was changed from 4:1 to 1:1. Twenty Japanese elderly subjects were initially given a SO diet for at least 6 mo (baseline period), a PO diet for 10 mo (intervention period), and then returned to the previous SO diet (washout period). ALA in the total serum lipid increased from 0.8 to 1.6% after 3 mo on the PO diet, but EPA and DHA increased in a later time, at 10 mo after the PO diet, from 2.5 to 3.6% and 5.3 to 6.4%, respectively (p<0.05), and then returned to baseline in the washout period. In spite of increases of serum n-3 fatty acids, the OxLDL concentration did not change significantly when given the PO diet. Body weight, total serum cholesterol, triacylglycerol, glucose, insulin and HbA1c concentrations, platelet count and aggregation function, prothrombin time, partial thromboplastin time, fibrinogen and PAI-1 concentration, and other routine blood analysis did not change significantly when given the PO diet. These data indicate that, even in elderly subjects, a 3 g/d increase of dietary ALA could increase serum EPA and DHA in 10 mo without any major adverse effects.

Aged↗

[Intravesical bacillus Calmette-Guerin instillation therapy for carcinoma in situ of the urinary bladder and prediction of effects by urinary cytologic examination].

PURPOSE: We investigated the effects of intravesical BCG instillations for carcinoma in situ (CIS) of the urinary bladder. And we have retrospectively analyzed the prediction of effects by fresh urinary cytologic examinations before instillation. MATERIAL AND METHODS: 33 patients were treated for bladder CIS (1991-1997) with a median follow-up of 30 months (range from 9 to 90 months). The patients (27 males and 6 females) ranged in age from 46 to 91 (average 71 years) and received 6 to 12 weekly BCG Tokyo 172 strain 80 mg instillations. They were divided into 3 groups based on tumor history: primary (9), secondary (15), concurrent (9). The prediction of effects were analyzed by scoring fresh urinary cytologic examinations before instillation. RESULTS: 22 cases (67%) were responded and they have remained free of disease for follow-up period. The statistic evaluation proved to show the significance between the effects of treatment and the sum of scoring (cellular appearance and existence of large nuclear cells). CONCLUSION: We confirmed the effects of this treatment. The prediction of effect of this treatment seemed to be indicative by fresh urinary cytologic examinations before instillation, especially cellular appearance and existence of large nuclear cells.

Adjuvants, Immunologic↗

[Long-term healing of excimer laser ablated rabbit corneas].

PURPOSE: To evaluate histologically and immunohistochemically long-term changes in rabbit cornea following excimer laser ablation. MATERIALS AND METHODS: Excimer laser keratectomy was performed on 12 eyes of 6 rabbits. The corneas were subjected to histological and immunohistochemical studies 2 and 3 years after treatment. FINDINGS: In all specimens, epithelial basement membrane of the ablated central cornea showed undulating and spike-pattern invasion into the epithelial side. The collagen lamellae showed irregularities in the subepithelial stroma. Immunohistochemically, the subepithelial stroma showed positive reaction to anti-type III collagen antibody. Staining for anti-IV collagen antibody showed a linear pattern in the epithelial basement membrane and was positive in the subepithelial stroma of the ablated central cornea. These positive reactions to anti-type III and type IV collagen antibodies in the stroma showed the localized presence of scar tissue in the ablated stroma. Antibody to type VII collagen showed weak reaction in the basement membrane, suggesting incomplete regeneration of anchoring fibrils. CONCLUSION: The healing process was still incomplete 3 years after excimer laser keratotomy in rabbits.

Animals↗

Normal structure and morphological effects of a corticosteroid on the peripatellar synovial membrane of the rat.

We observed the peripatellar synovial membranes of normal and betamethasone-treated rats with a scanning electron microscope (SEM), light microscope (LM) and transmission electron microscope (TEM). On the basis of SEM findings, the peripatellar synovial membrane is divided into two distinct regions; an upper, smaller, round, non-folded region which is covered with small excrescences, and a lower, larger, horseshoe-shaped region which has numerous folds consisting of cobblestone-like oval cells. These SEM findings were well confirmed by parallel LM and TEM observations of both regions. The synovial membrane of rats treated with intra-articular injections of betamethasone, an anti-inflammatory agent for rheumatoid arthritis, showed cell-sized excrescences on the surface of the non-folded regions. Under LM and TEM, the synovial membrane of treated rats showed an expansion of the intercellular spaces with collagen fibers, swelling of vacuoles in the cytoplasm in the type M synoviocytes, and increases in the number and elongation of microvilli of the type M synoviocytes. These observations suggest that repeated betamethasone injection therapy may result in unnecessary damage to the synovial membrane.

Animals↗

[Cystic renal cell carcinoma diagnosed as a simple cyst preoperatively with incidental renal tumor: a case report].

A 54-year-old man visited our hospital with right incidentally-found renal tumor detected by ultrasonography. Computed tomography, magnetic resonance imaging and angiography showed a small tumor, 1.5 cm in size, at the upper portion and a simple cyst, 4 cm in size, at the lower pole of the right kidney. We enucleated the small tumor and aspirated the cyst with outer part resection of the cyst wall. Pathological findings of the tumor showed renal cell carcinoma, alveolar type, common type, clear cell subtype, G1, pT1, INF-alpha. Microscopic appearance of the excised cyst wall also revealed sheets of renal cell carcinoma inside the wall. Therefore, two weeks after the first operation, we performed right radical nephrectomy. The resected specimen had severe inflammation without any evidence of residual tumor. Eight months after the nephrectomy, no recurrence has occurred.

Carcinoma, Renal Cell↗

Hexagonal bilayer structuring activity of linker chains of an annelid giant hemoglobin from the polychaete Perinereis aibuhitensis.

Preferential activity of linker chains to clamp submultiples to form hexagonal bilayer (HBL) assembly of the multisubunit hemoglobin (Hb) of the polychaete Perinereis aibuhitensis (approximately 3.4 MDa) was demonstrated. To understand the HBL assembly that should rely on structuring activity of each subunit, reassociation in response to combining isolated subunits was monitored using gel filtration, SDS-PAGE, and transmission electron microscopy. The isolation of each subunit L, T, and M (L, linker chains; T, disulfide-bonded trimer A-b-B; M, monomeric chain a) of Perinereis Hb was made simply by exposing Hb to pH 10.5, where Hb was completely dissociated into its subunits L, T, and M. As a result, it was concluded that (i) subunits T and M have strong affinity to form an intermediate complex, submultiple D, which is a dodecamer of globin chains, 3[a. A-b-B], (ii) addition of subunit L to submultiple D brings about the formation of whole molecule, similarly (iii) addition of subunit M to T+L forms the whole molecule, and (iv) addition of subunit T to M+L brings about the formation of the whole molecule, too. The results obtained lead us to conclude that linkers do function to clamp 12 submultiples D up to a whole molecule at the final step of formation of Perinereis Hb. In summary, linkers appeared to have high affinity for submultiple D, a little affinity for subunit T, but no affinity for subunit M at all. Thus linker chains were demonstrated to preferentially clamp submultiples D together to form the HBL disc of the whole molecule.

Animals↗

Carbohydrate gluing is a strategy for supramolecular clamping of submultiples in annelid extracellular multi-subunit hemoglobin.

A key to understanding the mysterious hierarchic organization of annelid multi-subunit extracellular hemoglobin (giant Hb, composed of 144 globin chains and about 36 nonheme chains called linkers) is knowing the role of linkers in holding together the entire two-tiered hexagonal form. Here, the effects of added monosaccharides on the dissociation of giant Hb from the marineworm Perinereis aibuhitensis were monitored using dynamic light scattering (DLS), transmission electron microscopy (TEM), and circular dichroism (CD) measurements. Changes in Stokes radius and more clearly the distribution analysis of the Hb based on the DLS measurements showed that Hb preferentially dissociates into hexagonal units (called submultiples), which was consistent with the results of TEM and CD measurements. The results thus show that linkers specifically "clamp" submultiples together to organize the two-tiered form through carbohydrate gluing. Thus, a submultiple behaves like an ordinary protein, whereas the intact Hb behaves like a miniature supramolecular system. This clamp model is plausible because it inherently involves catastrophe of the molecular stoichiometry at the two-tiered hexagonal formation level because carbohydrates are under posttranslational regulation and therefore contain structural ambiguity.

Animals↗

Early Breast Cancer Following Treatment of Myelodysplastic Syndrome: Report of a Case.

A 45-year-old woman was admitted to our hospital complaining of a mass in herleft breast. She had previously been diagnosed with myelodysplastic syndrome(MDS), a type of refractory anemia, based on bone marrow findings and chromosome analysis. She received a preoperative transfusion of fresh packed platelets and a recombinant human granulocyte colony-stimulating factor(rhG-CSF)injection. Left partial mastectomy and axillary lymph nodes dissection were performed to treat early breast cancer. Postoperatively, prophylactic radiotherapy of the residual breast and administration of medroxyprogesterone acetate(MPA)were performed because the tumor tissue was positive for progesterone receptors. She has remained clinically stable, with no evidence of recurrence, for more than three years to date. We report a rare case of breast cancer with MDS, treated with breast-conserving therapy. The strategy of pre- or postoperative platelet transfusion, rhG-CSFinjections, and hormonal therapy (MPA) appears to be suitable treatment for progesterone receptor(PgR)-positive breast cancer patients with MDS.

Journal Article↗

Human neuroblastoma demonstrating clonal evolution in vivo.

Neuroblastoma demonstrates various clinical behaviors, ranging from spontaneous regression to rapid progression regardless of the therapy used. To study the possibility that progression occurs in neuroblastoma through the accumulation of genetic aberrations, we analyzed the clonal constitution of the primary tumor and metastatic tumor samples from a stage-4 patient. Using cytofluorometry and FISH analyses, intratumor clonal heterogeneity was revealed. In the initial primary tumor sample, the nuclear DNA content indicated the coexistence of diploid and aneuploid clones, and the copy number of chromosome 1 varied from two to six. The chromosome 1 aneusomy population was composed of MYCN-amplified and 1p-deleted clones, whereas, in the chromosome 1 disomy population, coexistence of MYCN-amplified and non-amplified clones as well as 1p-deleted and 1p-intact clones was revealed. In the primary tumor after chemotherapy, the DNA-diploid component had become predominant, although the coexistence of MYCN-amplified and non-amplified clones could still be demonstrated in poorly- and well-differentiated tumor regions, respectively. This contrasted with the findings in the metastatic tumors, in which either diploid or aneuploid clone with MYCN amplification and 1p deletion dominated completely in each metastatic site. The findings suggest that the aneuploid clones had evolved from a diploid clone with MYCN amplification and a 1p deletion which, in turn, may have evolved from a diploid clone with neither MYCN nor 1p abnormality. This illustrates how various stages of multiple-step tumorigenesis may provide clues to a better understanding of the clinical heterogeneity of neuroblastoma.

Child, Preschool↗

Immunohistochemical localization of arginase II and other enzymes of arginine metabolism in rat kidney and liver.

Arginine is a precursor for the synthesis of urea, polyamines, creatine phosphate, nitric oxide and proteins. It is synthesized from ornithine by argininosuccinate synthetase and argininosuccinate lyase and is degraded by arginase, which consists of a liver-type (arginase I) and a non-hepatic type (arginase II). Recently, cDNAs for human and rat arginase II have been isolated. In this study, immunocytochemical analysis showed that human arginase II expressed in COS-7 cells was localized in the mitochondria. Arginase II mRNA was abundant in the rat small intestine and kidney. In the kidney, argininosuccinate synthetase and lyase were immunostained in the cortex, intensely in proximal tubules and much less intensely in distal tubules. In contrast, arginase II was stained intensely in the outer stripes of the outer medulla, presumably in the proximal straight tubules, and in a subpopulation of the proximal tubules in the cortex. Immunostaining of serial sections of the kidney showed that argininosuccinate synthetase and arginase II were colocalized in a subpopulation of proximal tubules in the cortex, whereas only the synthetase, but not arginase II, was present in another subpopulation of proximal tubules. In the liver, all the enzymes of the urea cycle, i.e. carbamylphosphate synthetase I, ornithine transcarbamylase, argininosuccinate synthetase and lyase and arginase I, showed similar zonation patterns with staining more intense in periportal hepatocytes than in pericentral hepatocytes, although zonation of ornithine transcarbamylase was much less prominent. The implications of these results are discussed.

Animals↗

Regulation of the urea cycle enzyme genes in nitric oxide synthesis.

Nitric oxide (NO) is synthesized from arginine by nitric-oxide synthase (NOS), and citrulline that is generated can be recycled to arginine by argininosuccinate synthase (AS) and argininosuccinate lyase (AL). Rats were injected with bacterial lipopolysaccharide (LPS) and expression of the inducible isoform of NOS (iNOS), AS and AL was analysed. In RNA blot analysis, iNOS mRNA was induced by LPS in the lung, heart, liver and spleen, and less strongly in the skeletal muscle and testis. AS and AL mRNAs were induced in the lung and spleen. Kinetic studies showed that iNOS mRNA increased rapidly in both spleen and lung, reached a maximum 2-5 h after the treatment, and decreased thereafter. On the other hand, AS mRNA increased more slowly and reached a maximum in 6-12 h (by about 10-fold in the spleen and 2-fold in the lung). AL mRNA in the spleen and lung increased slowly and remained high up to 24 h. In immunohistochemical analysis, macrophages in the spleen that were negative for iNOS and AS before LPS treatment were strongly positive for both iNOS and AS after this treatment. As iNOS, AS and AL were co-induced in rat tissues and cells, citrulline-arginine recycling seems to be important in NO synthesis under the conditions of stimulation. Arginine is a common substrate of NOS and arginase. Rat peritoneal macrophages were cultured in the presence of LPS and expression of iNOS and livertype arginase (arginase I) was analysed. mRNAs for iNOS and arginase I were induced by LPS in a dose-dependent manner. iNOS mRNA appeared 2 h after LPS treatment and increased up to a near-maximum at 8-12 h. On the other hand, arginase I mRNA began to increase after 4 h with a lag time and reached a maximum at 12 h. Immunoblot analysis showed that iNOS and arginase I proteins were also induced. Induction of iNOS and arginase I mRNAs were also observed in LPS-injected rats in vivo. Thus, arginase I appears to have an important role in downregulating NO synthesis in murine macrophages by decreasing the availability of arginine. A cDNA for human arginase II, an arginase isozyme, was isolated. A polypeptide of 354 amino acid residues including the putative NH2-terminal presequence for mitochondrial import was predicted. It was 59% identical with arginase I. mRNA for human arginase II was present in the kidney and other tissues but was not detected in the liver. Arginase II mRNA was co-induced with iNOS mRNA in murine macrophage-like RAW 264.7 cells by LPS. This induction was enhanced by dexamethasone and dibutyrul cAMP, and was prevented by interferon-gamma. These results indicate that NO synthesis is regulated by arginine-synthesizing and -degrading enzymes in a complicated manner.

Amino Acid Sequence↗

Effects of parent Shamo cocks on the histochemical properties of M. iliotibialis lateralis and M. supracoracoideus on their crossbred broilers.

1. Four Shamo (a Japanese game bird) cocks showing different characteristics in the histochemical properties of M. iliotibialis lateralis (ITL) were crossed with White Rock hens to produce male and female crossbred broilers of the 4 lines (90 d of age). Normal broilers (56 d) were used, for comparison. 2. Histochemical properties of ITL and M. supracoracoideus (SC) were compared among the crossbred lines and normal broilers. Myofibres were divided into Types II R, II I and II W showing high, moderate and low reduced nicotinamide adenine dinucleotide dehydrogenase (NADH-DH) activities, respectively. 3. In the ITL of the crossbred cockerels, the percentage of Type II R and II I fibres decreased and conversely Type II W increased in comparison to those in the Shamo. 4. Sex differences of the histochemical properties were recognised only in the ITL of the crossbred, in which the percentage of Type II R fibres was greater in the male. 5. The different characteristics of the parent Shamo cocks were reproduced only in the different fibre type composition of the ITL muscle in the crossbred cockerels. 6. The histochemical features of fibre type seemed to develop with bird age, particularly subsarcolemmal accumulation of formazan granules (indicating high NADH-DH activity) in Type IIR fibres. 7. Breed, line, sex and age differences in the histochemical properties were demonstrated clearly in ITL but not in SC.

Age Factors↗

Gastric ulcer, atrophic gastritis, and intestinal metaplasia caused by Helicobacter pylori infection in Mongolian gerbils.

BACKGROUND: Helicobacter pylori infection is associated with gastroduodenal disease in humans. In this study we aimed to show this relationship directly in Mongolian gerbils. METHODS: The animals were challenged orally with H. pylori and killed 1, 2, 3, and 6 months after inoculation for histologic and anti-H. pylori antibody titer examination. RESULTS: The spiral bacteria were observed in the mucus and gastric pits of all infected animals. A severe infiltration of the lamina propria by polymorphonuclear and mononuclear cells was seen 1 month after H. pylori inoculation. The submucosa was infiltrated by mainly mononuclear cells with formation of lymphoid follicles. Erosion of the gastric mucosa appeared soon after inoculation, whereas gastric ulcers, gastritis cystica profunda, and atrophy with goblet cell metaplasia occurred between 3 and 6 months after inoculation. In the duodenal mucosa a mild inflammatory cell infiltration with ballooning and diminished number of duodenal glands was seen. The IgG anti-H. pylori antibody titer increased gradually after 2 months of inoculation. CONCLUSIONS: Since the gastritis, gastric ulcers, atrophic gastritis, and intestinal metaplasia that developed in Mongolian gerbils were similar to those observed in humans, this model may be useful to study the therapy of gastric ulcer and, with a longer observation period, to confirm a possible relationship between H. pylori and malignancy.

Animals↗

Chemoprevention of N-nitroso-N-methylurea-induced rat mammary carcinogenesis by soy foods or biochanin A.

We examined the effects of soybeans, a soy product (miso) and biochanin A, an isoflavone derivative, on N-nitroso-N-methylurea (MNU)-induced rat mammary carcinogenesis. Seven-week-old female CD/Crj rats received a single i.v. dose (40 mg/kg body weight) of MNU. After administration of MNU, rats were fed diet containing 0% (control), 2% or 10% soybeans, or 10% miso as a soy-supplemented diet, or 10 or 50 mg/kg biochanin A. All rats were observed for 18 weeks after MNU administration. At 18 weeks, the multiplicity (mean tumors/rat) of palpable mammary tumors was significantly decreased in the 10% soybean (1.1) and 10% miso (1.2) diet groups compared to the control (2.2) (P<0.05, respectively). In the biochanin A-supplemented diet groups, the incidence (percentage of rats with tumors) was significantly decreased in the 50 mg/kg (32%) diet group compared to the control (80%) (P<0.01), and the multiplicity was significantly decreased in both the 10 mg/kg (0.7) and 50 mg/kg (0.5) diet groups compared to the control (2.2) (P<0.01 and P<0.001, respectively). The proliferative cell nuclear antigen labeling index of mammary tumors was significantly decreased in both biochanin A-supplemented diet groups compared to the control. The present results indicate that soybeans, miso, and biochanin A are useful for the prevention of mammary cancer.

Animals↗

Chemoprevention of N-nitroso-N-methylurea-induced rat mammary cancer by miso and tamoxifen, alone and in combination.

We examined the effects of a Japanese fermented soybean product, miso, and tamoxifen (TAM), alone and in combination, on N-nitroso-N-methylurea (MNU)-induced rat mammary cancer. Seven-week-old female CD/Crj rats received a single i.v. dose (50 mg/kg body weight) of MNU. After administration of MNU, the rats were divided into 4 groups: regular diet (control), 10% miso diet, regular diet + TAM, and 10% miso diet + TAM. TAM was implanted s.c. in the form of pellets containing 2.5 mg at the same time as MNU was administered. All rats were observed for 18 weeks after MNU administration. Incidence (percentage of rats with tumors) and multiplicity (mean tumors/rat) of mammary tumors were 91% and 4.5 in the control, 77% and 2.4 (P < 0.05) in the 10% miso group, 68% and 1.4 (P <0.01) in the TAM group, and 10% (P < 0.0001 or less) and 0.2 (P < 0.0001) in the 10% miso + TAM group. In the second experiment, the effect of the combination of miso and TAM on established rat mammary tumors was investigated. When the mammary tumors induced by MNU reached 10 to 25 mm, the rats were divided into 3 treatment groups: regular diet, regular diet + TAM, and 10% miso diet + TAM. At 6 weeks after the start of treatment, the mean tumor size in the control and TAM groups was 160% and 141% of the pretreatment value, but a decrease of 85% of the pretreatment value was produced by the combination of miso and TAM, and this was significantly different from both the control and TAM groups (P < 0.01) and P < 0.05, respectively). These results indicate that miso is useful in protecting against mammary cancer and it can be expected to have a potent antitumor effect, especially when used in combination with TAM.

Animals↗

Serological assessment of the early response to eradication therapy using an immunodominant outer membrane protein of Helicobacter pylori.

Eradication of Helicobacter pylori infection cures gastritis and prevents recurrence of peptic ulcers. Endoscopy is usually used to evaluate the effectiveness of eradication therapy. We designed a new noninvasive assay system for the early evaluation of eradication of H. pylori infection in which a crude H. pylori outer membrane protein preparation (HPOmp) is used as an antigen, and we determined the sensitivity and specificity of the serological assay system. Immunoblot analysis showed that anti-HPOmp antibodies reacted to a protein with a molecular mass of approximately 29 kDa. In those patients who responded to therapy, the anti-HPOmp immunoglobulin G (IgG) titers measured by enzyme-linked immunosorbent assay (ELISA) at 1 month after the end of therapy were significantly lower than those before treatment (34.8% reduction; P < 0.001), and the posttreatment reduction in the antibody titer was significantly greater than that of the titer measured with a commercially available anti-H. pylori IgG ELISA (34.8% versus 16.1%; P < 0.001). When a 25% reduction of anti-HPOmp IgG titer at 1 month after the end of treatment was taken as the cutoff value for H. pylori eradication, the sensitivity and specificity of our new assay were 75% (51 of 68 treatment responders) and 96% (22 of 23 nonresponders), respectively. Our results indicate that the novel serological test with HPOmp might be a clinically useful tool for assessment of eradication of H. pylori.

Adolescent↗

Distinct arginase isoforms expressed in primary and transformed macrophages: regulation by oxygen tension.

Experiments were performed to identify arginase isoforms expressed in primary and transformed rodent macrophages and to determine the molecular mechanisms for the previously observed increase in arginase activity in macrophages cultured in hypoxia or anoxia. Results demonstrate the following: 1) mRNA and protein for hepatic-type AI arginase are expressed in primary cultures of rat and mouse peritoneal macrophages and are enhanced seven- and nine-fold, respectively, by lipopolysaccharide (LPS). 2) mRNA for extrahepatic-type AII arginase is constitutively expressed in mouse, but not rat, peritoneal macrophages and is detected in RAW264.7 cells after LPS treatment; neither J774A.1 nor P388D1 cells contain arginase mRNA. 3) AI arginase mRNA, arginase activity in cell lysates, and L-arginine flux through arginase in intact cells are all increased in rat wound-derived and mouse peritoneal macrophages by hypoxic or anoxic culture; AII arginase mRNA is, in contrast, suppressed > 50% by O2 deprivation. 4) Expression of the L-arginine transporter mCAT-2 is increased greater than twofold by reduced O2 culture. These results demonstrate substantial variability in arginase isoform expression among primary and transformed rodent macrophages. They also identify AI and AII arginase and the mCAT-2 L-arginine transporter as O2-regulated genes.

Amino Acid Transport Systems, Basic↗