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T Ohta

Publications and source records attributed to T Ohta.

At least 415 records · Page 23Linked to original sources

[Recent development and clinical application of bone mineral measurements].

Over the past decade, methodologies for the assessment of bone mineral density have markedly progressed, so that any sites of the skeleton now can be measured with high degree of accuracy and precision with safety. The number of devices distributed in Japan rapidly increased for the last 5 years and the total number installed nationwide reached over 7000 as with 1998. There are variety of techniques: microdensitometry (MD) or radiographic absorptiometry (RA), single X-ray absorptiometry (SXA), dual X-ray absorptiometry (DXA), quantitative CT (QCT), peripheral QCT, and quantitative ultrasonometry (QUS). There are, however, no such single technique as to fulfill the entire clinical requirements, since the time of initiation of bone loss, and the speed of bone loss are quite different from site to site of the skeleton, so that the correlations of bone density measured by each technique are not sufficiently high (gamma = 0.5-0.8) to predict BMD of other bones by measuring one bone. Since the relatively large amounts of data on the prediction of fracture (hip, spine and others) by these techniques have been accumulated, a specific guideline regarding the appropriate application of these techniques, including multiple combination measurements, should be established based on the worldwide consensus.

Bone Density↗

Effects of continuous alendronate treatment on bone mass and mechanical properties in ovariectomized rats: comparison with pamidronate and etidronate in growing rats.

Alendronate is a potent inhibitor of bone resorption. To investigate the relationship between antiresorptive activity and bone-related side effects, we studied the effect of 2 months of daily alendronate (0.04, 0.2, 1.0 or 5.0 mg/kg/day) treatment on the strength of the femoral shaft and neck and on the bone mass of ovariectomized rats. The p.o. administration regimen began immediately after ovariectomy at 6 weeks of age, and the results were compared with pamidronate (0.2, 1.0 or 5.0 mg/kg/day) or etidronate (5.0, 25.0 or 125.0 mg/kg/day) treatment. In the femoral epiphysis and neck, a preventive effect of alendronate on loss of bone mineral density was observed at the dose of 1.0 mg/kg. The alendronate-treated group did not show significant alteration of the breaking load or the cross-sectional shape of the femoral midshaft. Similar results were obtained in the femoral neck strength and femoral neck geometry. In histomorphometric analysis of tibial metaphyses, alendronate inhibited the ratio of osteoid volume to tissue volume and the mineral apposition rate at a dose of 0.2 mg/kg compared with the ovariectomized control. In contrast, etidronate tended to increase osteoid volume/bone volume at 125 mg/kg. From these results, we conclude that p.o. alendronate-treatment prevented the decrease in bone mineral density and maintained the mechanical properties of bone after ovariectomy without impairing of bone mineralization in growing rats.

Alendronate↗

Evolution by nearly-neutral mutations.

Genetic systems are complex and interact at various levels; among amino acids or nucleotide sites, among gene products, and among regulatory regions and proteins. Patterns of synonymous and nonsynonymous substitutions of mammalian genes indicate that nonsynonymous substitutions are nearly neutral, coming from interactions among amino acids. As an interactive system, the NK model of Kauffman was analysed. This model assumes that each amino acid makes a fitness contribution that depends upon the amino acid and upon K other amino acids among the N that make the protein. Through simulations, it was found that there are numerous nearly-neutral mutations in this model, and that evolution is rapid in small populations and slow in large ones. The system moves on the rugged fitness landscape by mutation, random genetic drift and selection. Small populations have more chance to attain novel genetic systems than large ones because of larger effects of random drift, but the chance of extinction becomes greater.

Animals↗

[Non-Hodgkin's lymphoma of the pancreas with elevated serum CA 19-9 and SPAN-1].

A 55-year-old woman was admitted because of progressive jaundice. Blood examination on admission revealed markedly elevated serum levels of CA19-9 and SPAN-1. Abdominal computed tomography revealed a large tumor in the head of the pancreas. Although the patient's jaundice and elevated CA19-9 decreased after percutaneous franshepatic cholangio-drainage, her SPAN-1 level remained elevated. Open biopsy of the pancreatic tumor revealed non-Hodgkin's lymphoma (NHL) (diffuse medium, B cell type), Complete remission was obtained after one course of CHOP therapy. This case suggests that pancreatic tumor with elevated serum CA19-9 and SPAN-1 levels may involve NHL, and may be curable with chemotherapy.

Antigens, Neoplasm↗

[Indications for and operative outcome of hepato-pancreatoduodenectomy in the treatment of carcinoma of the gallbladder].

The indications for hepatopancreatoduodenectomy (HPD) are still controversial. Opinion is divided especially concerning the need to perform pancreatoduodenectomy (PD) for lymph node dissection, with some investigators claiming that the peripancreatic lymph nodes can be dissected adequately without PD. Detailed studies of resected specimens in our department have shown that preservation of the head of the pancreas carries an increased risk of leaving behind cancer cells in small lymph nodes and ducts remaining in the peripancreatic region. We therefore perform HPD in all patients at increased risk for metastasis to peripancreatic lymph nodes, as indicated by subserosal invasion with distinct evidence of lymph node involvement, tumor location at the neck or both fundus and body of the gallbladder, and serosa exposure to tumor irrespective of tumor location and lymph node involvement. When the outcome was compared between HPD and hepatic resection plus dissection of peripancreatic lymph nodes (HR), we found that the operative results of HPD were poor with extended lobectomy plus PD because of high rates of postoperative complications and operative mortality. However, the results have gradually improved with the introduction of percutaneous transehepatic portal embolization and advances in perioperative care. Although overall there is no difference between HPD and HR in long-term outcome, the outcome with HPD is significantly better in node-positive patients and patients without hepatoduodenal ligament involvement, there by demonstrating the value of performing peripancreatic lymph node dissection by PD. However, there is no difference between HPD and HR in patients with hepatoduodenal ligament involvement, and attempts to develop other new and effective means of treatment should continue.

Gallbladder Neoplasms↗

The meaning of near-neutrality at coding and non-coding regions.

The nature of weak selection differs between coding and non-coding regions. Coding regions contain genetic information, whereas most non-coding regions do not have any information. Genetic information may be regarded as interaction systems, and the NK model of Kauffman was analysed. This model assumes that each amino acid makes a fitness contribution that depends on the amino acid and on K other amino acids among the N that make the protein. Through simulations, it was found that there are numerous nearly-neutral mutations under this model. Therefore, evolution is rapid in small populations, and slow in large populations. The variance of the evolutionary rate is not quite as large as data indicate under the model, and additional factors, such as environmental change or population-size fluctuation, need to be considered. Weak selection at non-coding regions may come from chromosome organization, and may be regional in character, which differs from that at coding regions. The problem of genetic load is thought to disappear in these circumstances.

Gene Frequency↗

Le(y) glycolipid acts as a co-factor for tumor procoagulant activity.

We have generated a monoclonal antibody (MAb), FS01, which inhibits the procoagulant activity (CCA-1) produced by a human squamous cell carcinoma cell line, LK52. Expression of the antigen recognized by FS01 MAb in various cancer cell lines correlated well with the procoagulant activities of the expressing cell lines. Our objective was to characterize the molecule reacting with FS01 MAb and to analyze its involvement in the CCA-1 procoagulant activity. The molecule was identified as a glycolipid and found to be involved in the procoagulant activity because both procoagulant activity and reactivity to FS01 MAb were lost after endoglycoceramidase treatment of CCA-1. Furthermore, FS01 MAb recognized the Lewis Y (Le[y]) antigen. To confirm the involvement of a glycolipid incorporating the Le(y) antigen in the procoagulant activity, we attempted to purify CCA-1 from LK52 culture supernatant. In one of the purification steps, a fraction containing low procoagulant activity (CCA-1p) separated from the Le(y)-positive fraction (CCA-1c). Although CCA-1c alone did not show procoagulant activity, the procoagulant activity of CCA-1p was augmented by CCA-1c and this augmentation was inhibited by FS01 MAb. Furthermore, CCA-1c enhanced the procoagulant activity of 33 cell lines tested as well as CCA-1p. In addition, purified Le(y) glycolipid from canine intestine augmented the procoagulant activity of CCA-1p, and this augmentation also could be inhibited by FS01 MAb. We conclude that Le(y) glycolipid is a co-factor for the procoagulant activity derived from cancer cells.

Animals↗

Changes in intracellular Na+ concentration evoked by nicotinic receptor activation in the guinea-pig adrenal chromaffin cells.

Using the whole-cell voltage clamp technique and microfluorometry with sodium-binding benzofuran isophthalate (SBFI), a nicotine-induced inward current and increase in the intracellular Na+ concentration ([Na+]in) were examined simultaneously in guinea-pig adrenal chromaffin cells. The increase in [Na+]in expected from the time-integrated inward current was well correlated with that of [Na+]in measured with SBFI. The ratio of the expected [Na+]in to the measured [Na+]in was 0.64 at -85mV and decreased with increasing holding potentials. The decay time constant of the increased [Na+]in was not affected by ouabain. It is concluded that the Na+ entering the cell is diffusable in about 60% cell volume without fast buffering mechanisms and is eliminated by the exchange of Na+ between the pipette solution and cell interior under the patch clamp condition.

Animals↗

Co-mutagenicity of glyco- and tauro-deoxycholic acids in the Ames test.

Mutagenicity and co-mutagenicity of glyco- and tauro-deoxycholic acids (GDCA and TDCA), which are abundant in human bile, were examined by the Ames test. The two chemicals were not mutagenic for themselves to Salmonella typhimurium TA98 and TA100, with and without S9 mix. They enhanced, however, the mutagenic activities of the pro-mutagens, 2-aminoanthracene (2AA) and benzo[a]pyrene (BaP), for both TA98 and TA100 with S9 mix. They were more co-mutagenic for the pro-mutagens on TA98 than on TA100. On TA98, the mutagenic activities of 2AA with GDCA (5 mumol/plate) and with TDCA (5 mumol/plate) were 9.7-fold and 11.8-fold as high as that of the corresponding control (2AA only), respectively. BaP with GDCA (2.5 mumol/plate) and with TDCA (2.5 mumol/plate) showed 2.8-fold and 3.0-fold increases over the corresponding control level (BaP only), respectively. It is hence concluded that GDCA and TDCA may enhance the activity of some mutagens existing in bile.

Animals↗

In vivo genotoxicity of heterocyclic amines detected by a modified alkaline single cell gel electrophoresis assay in a multiple organ study in the mouse.

We used a modification of the alkaline single cell gel electrophoresis (SCG) (Comet) assay to test the in vivo genotoxicity of 6 heterocyclic amines, Trp-P-1 (25 mg/kg), Trp-P-2 (13 mg/kg), IQ (13 mg/kg), MeIQ (13 mg/kg), MeIQx (13 mg/kg) and PhIP (40 mg/kg), in mouse liver, lung, kidney, brain, spleen, bone marrow and stomach mucosa. Mice were sacrificed 1, 3, and 24 h after intraperitoneal injection. Trp-P-2, IQ, MeIQ, and MeIQx yielded statistically significant DNA damage in the stomach, liver, kidney, lung and brain; Trp-P-1 in the stomach, liver and lung; and PhIP in the liver, kidney and brain. None of the heterocyclic amines induced DNA damage in the spleen and bone marrow. Our results suggest that the alkaline SCG assay applied to multiple organs is a good way to detect organ-specific genotoxicity of heterocyclic amines in mammals.

Amines↗

Structural and functional characterization of homo-oligomeric complexes of alpha and beta chaperonin subunits from the hyperthermophilic archaeum Thermococcus strain KS-1.

To elucidate the function of group II chaperonin, the gene for the chaperonin from the hyperthermophilic archaeum Thermococcus strain KS-1 was cloned and sequenced. Two distinct genes coding for chaperonin subunits, designated alpha and beta, were obtained, and their deduced amino acid sequences are highly homologous to those of group II chaperonins from other sources. The alpha and beta subunits were individually expressed in Escherichia coli. Both of the recombinant subunits assemble to constitute the homo-oligomeric double-ring complexes, which are prone to form large aggregates. The alpha aggregate is dissociated into the typical chaperonin ring complex by incubation in buffer containing 15% (v/v) methanol, while the beta aggregate cannot be dissociated. At high temperature, both of the recombinant complexes have weak ATPase activities. They are able to arrest refolding of a chemically denatured thermophilic enzyme in the absence of ATP, and refolding is resumed when ATP is supplemented. These results suggest that homo-oligomeric complexes of the archaeal chaperonin have activity.

3-Isopropylmalate Dehydrogenase↗

Purification and characterization of the human interleukin-18 receptor.

Interleukin (IL)-18 was identified as a molecule that induces IFN-gamma production and enhances NK cell cytotoxicity. In this paper, we report upon the purification and characterization of human IL-18 receptor (hIL-18R). We selected the Hodgkin's disease cell line, L428, as the most strongly hIL-18R-expressing cell line based on the results of binding assays. This binding was inhibited by IL-18 but not by IL-1beta. The dissociation constant (Kd) of 125I-IL-18 binding to L428 cells was about 18.5 nM, with 18,000 binding sites/cell. After immunizing mice with L428 cells and cloning, a single monoclonal antibody (mAb) against hIL-18R was obtained (mAb 117-10C). Sequentially, hIL-18R was purified from 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS)-extracted L428 cells by wheat germ lectin-Sepharose 4B chromatography and mAb 117-10C-Sepharose chromatography. The internal amino acid sequences of hIL-18R all matched those of human IL-1 receptor-related protein (IL-1Rrp), the ligand of which was unknown to date. When expressed in COS-1 cells, the cDNA of IL-1Rrp conferred IL-18 binding properties on the cells and the capacity for signal transduction. From these results, we conclude that a functional IL-18 receptor component is IL-1Rrp.

Amino Acid Sequence↗

Requirement of the caspase-3/CPP32 protease cascade for apoptotic death following cytokine deprivation in hematopoietic cells.

Hematopoietic cytokines transduce cell survival signals, which are distinct from the signals necessary for the stimulation of DNA synthesis. Recently, the Ras and phosphatidylinositol 3-kinase pathways have been shown to play important roles in preventing apoptosis in various cell types, e.g. hematopoietic cells and neuronal cells. Withdrawal of cytokine(s), in turn, results in rapid inactivation of these survival pathways and eventually leads to cell death accompanied by the hallmarks of apoptosis. However, the mechanism of cell death caused by cytokine deprivation has not been fully elucidated. In this study, we demonstrate that caspase-3/CPP32, a member of the caspase/interleukin-1beta-converting enzyme family, is activated upon interleukin (IL)-3 deprivation in IL-3-dependent cells as well as IL-2 deprivation in IL-2-dependent cells. In addition, poly(ADP-ribose) polymerase, a cellular substrate for the caspase family proteases, was degraded into apoptotic fragments in both cell lines after cytokine removal. Furthermore, inhibition of a caspase family protease by synthetic peptides suppressed apoptotic death. These results indicate that the activation of a caspase-like protease(s) is required for the progression of apoptosis following cytokine deprivation. However, readdition of IL-3 did not restore the proliferative potential of the cells that survived in the presence of the peptide inhibitor after IL-3 depletion. Therefore, cellular commitment to apoptosis appears to precede the activation of a caspase-like protease(s).

Animals↗

Simple detection of in vivo genotoxicity of pyrimethamine in rodents by the modified alkaline single-cell gel electrophoresis assay.

We tested the genotoxicity of pyrimethamine in 5 mouse and rat organs (liver, lung, kidney, spleen, and bone marrow) using a modified alkaline single-cell gel electrophoresis (SCG) (Comet) assay. Mice and rats were sacrificed 1, 3, 6, and 24 h after oral administration of the drug at 50 and 120 mg/kg, respectively. Nuclei were isolated from each tissue and evaluated for DNA migration. Pyrimethamine induced DNA damage in cells of the liver, kidney, and lung in both species. For mice, DNA damage persisted in the liver for 24 h, while it peaked in the lung and kidney at 6 and 24 h, respectively. For rats, DNA damage in the liver peaked at 1 h and returned to almost control level at 24 h. Genotoxicity in the spleen was only observed in mice. Our results suggest that the SCG technique, using isolated nuclei can be applied to rats and mice and that the optimal sampling time is different for different organs and species.

Animals↗

Inhibitory effects of HepG2 cell-derived apolipoprotein A-I-containing lipoproteins on cholesteryl ester accumulation in macrophages.

We investigated the mechanisms of inhibitory effects on foam cell formation of apolipoprotein A-I-containing lipoproteins secreted by HepG2 cells (HepG2-HDL) using mouse peritoneal macrophages. When macrophages were incubated with acetylated low-density lipoprotein (acetyl-LDL) in the presence of HepG2-HDL, cholesterol ester (CE) accumulation in cells was reduced by 63%. This inhibitory capacity was almost similar to that of plasma high-density lipoprotein (HDL). When macrophages were converted to foam cells with acetyl-LDL and then reacted with HepG2-HDL or plasma HDL, the HDL-induced CE reduction was 2.2-fold greater than HepG2-HDL. Similar results were obtained using apo E-free HepG2-HDL. Since the inhibitory effect of HDL on acetyl-LDL-induced CE accumulation in macrophages is due largely to its cholesterol efflux capacity, these results suggest the presence of an additional mechanism for the inhibition of CE accumulation by HepG2-HDL. To investigate the mechanism, acetyl-LDL was reisolated from HepG2-HDL by Sephacryl S-300 gel filtration after incubation in a cell-free system. Reisolated acetyl-LDL showed a significant reduction in electrophoretic mobility. The extent of CE accumulation by reisolated acetyl-LDL was reduced by 20% compared with control acetyl-LDL. Moreover, its endocytic degradation by macrophages was reduced by 28%. HepG2-HDL also inhibited macrophage degradation of acetyl-LDL as well as oxidized LDL, a likely atherogenic lipoprotein. This inhibitory effect was ascribed to the HepG2-HDL subfraction containing pre-beta HDL. Our results indicated that apo A-I-containing lipoproteins as a physiological model of nascent HDL may inhibit foam cell formation by reducing ligand activity of atherogenic lipoproteins. These data possibly suggest inhibitory function of nascent HDL for the formation of foam cells in vivo.

Animals↗

Interleukin-18 induces activation and association of p56(lck) and MAPK in a murine TH1 clone.

Interleukin-18 (IL-18) was identified as an inducer of interferon-gamma (IFN-gamma) production by stimulated T cells. In this study, we used an ovalbumin-responsive murine Th1 clone (OVA#4), in which DNA synthesis was reportedly enhanced after IL-18 treatment in the presence of a non-mitogenic TCR/CD3 stimulus, to examine signal transduction pathways. In the presence of the stimulus, IL-18 induced the appearance of tyrosine-phosphorylated proteins and herbimycin A inhibited DNA synthesis. It is suggested that protein tyrosine kinase (PTK) mediated signaling is induced by IL-18. Specifically, IL-18 induced phosphorylation of phosphorylates p56(lck) (LCK) and mitogen-activated protein kinase (MAPK). IL-18 alone induced the kinase activities of both LCK and MAPK, and the activities were increased by the TCR/CD3 stimulus. Simultaneously, IL-18 induced the association of LCK with MAPK and this was also increased by the TCR/CD3 stimulus. The activation of the LCK-MAPK pathway correlated with enhanced DNA synthesis in OVA#4 cells. These results suggest that the LCK-MAPK pathway is involved in IL-18 signaling and that IL-18 may play an important role in modification of TCR/CD3-mediated response.

Animals↗

Measurement of intracellular Na+ concentration by a Na+-sensitive fluorescent dye, sodium-binding benzofuran isophthalate, in porcine adrenal chromaffin cells--usage of palytoxin as a Na+ ionophore.

Palytoxin was found to equilibrate sodium ions (Na+) across the cell membrane much faster than dose gramicidin, which has been frequently used to calibrate the intracellular Na+ concentration ([Na+]in), in cells treated with a Na+-sensitive fluorescent dye, sodium-binding benzofuran isophthalate (SBFI). Palytoxin was capable of equilibrating Na+ in cells treated with SBFI-acetoxymethyl ester (SBFI-AM) and in voltage-clamped cells loaded with SBFI through a patch pipette. Nicotine caused a dose-dependent increase in ([Na+]in) in porcine adrenal chromaffin cells treated with SBFI-AM and caused a simultaneous increase in [Na+]in and inward current in the voltage-clamped cells loaded with SBFI. Palytoxin has an advantage of calibrating ([Na+]in) in a shorter time than dose gramicidin because of its powerful ionophoretic activity.

Acrylamides↗