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Biomedical subjects

T Mochizuki

Publications and source records attributed to T Mochizuki.

At least 91 records · Page 5Linked to original sources

Oncogenic Ras triggers cell suicide through the activation of a caspase-independent cell death program in human cancer cells.

To prevent neoplasia, cells of multicellular organisms activate cellular disposal programs such as apoptosis in response to deregulated oncogene expression, making the suppression of such programs an essential step for potentially neoplastic cells to become established as clinically relevant tumors. Since the mutation of ras proto-oncogenes, the most frequently mutated proto-oncogenes in human tumors, is very rare in some tumor types such as glioblastomas and gastric cancers, we hypothesized that mutated ras genes might activate a cell death program that cannot be overcome by these tumor types. Here we show that the expression of oncogenically mutated ras gene induces cellular degeneration accompanied by cytoplasmic vacuoles in human glioma and gastric cancer cell lines. Cells dying as a result of oncogenic Ras expression had relatively well-preserved nuclei that were negative for TUNEL staining. An immunocytochemical analysis demonstrated that the cytoplasmic vacuoles are derived mainly from lysosomes. This oncogenic Ras-induced cell death occurred in the absence of caspase activation, and was not inhibited by the overexpression of anti-apoptotic Bcl-2 protein. These observations suggested that oncogenic Ras-induced cell death is most consistent with a type of programmed cell death designated 'type 2 physiological cell death' or 'autophagic degeneration', and that this cell death is regulated by a molecular mechanism distinct from that of apoptosis. Our findings suggest a possible role for this non-apoptotic cell death in the prevention of neoplasia, and the activation of the non-apoptotic cell death program may become a potential cancer therapy complementing apoptosis-based therapies. In addition, the approach used in this study may be a valuable way to find genetically-regulated cell suicide programs that cannot be overcome by particular tumor types.

Brain Neoplasms↗

Validity of 99mTc-DMSA renal uptake by planar posterior-view method in children.

Renal uptake of 99mTc-DMSA has been quantified by various methods. The aim of this study is to obtain a normal value for 99mTc-DMSA renal uptake calculated by the posterior view method and age variation, and to assess its clinical validity. Scintigrams of 238 children (0-12 years) with 99mTc-DMSA were reviewed. All the children had a clinical history of primary vesicoureteral reflux and/or neurogenic bladder, ureteral or urethral anomalies. Their kidneys were divided into two groups, "normal" and "abnormal" according to their scintigraphic findings and split renal functions. Percent renal uptake per injected dose (% RU) was quantitated from planar images at 2 hours after injection of an age-adjusted dose (26-95 MBq) of 99mTc-DMSA. Calculated total % RU, individual % RU of the right and left kidneys (mean +/- sd) in patients with normal kidneys were 40.7 +/- 5.0%, 20.2 +/- 3.0%, 20.4 +/- 2.7%, respectively. There was no significant correlation between % RU and age (r = 0.231). Longitudinal variation in the % RU in 9 patients ranged from 1.2% to 18%. Our conventional method for quantifyng % RU is simple, practical and feasible in routine clinical practice, especially for children under follow up.

Adolescent↗

An alternative approach to estimation of the brain perfusion index for measurement of cerebral blood flow using technetium-99m compounds.

Cerebral blood flow (CBF) has been quantified non-invasively using the brain perfusion index (BPI) determined from radionuclide angiographic data generated by technetium-99m hexamethylpropylene amine oxime( )((99m)Tc-HMPAO) or technetium-99m ethyl cysteinate dimer( )((99m)Tc-ECD). The BPI is generally calculated using graphical analysis (GA). In the present study, BPI was measured using spectral analysis (SA), and its usefulness evaluated in comparison with GA. The BPI was calculated from the sum of spectral data obtained by SA. We applied this method to radionuclide angiographic data collected from the bilateral brain hemispheres of 20 patients with various brain diseases using (99m)Tc-HMPAO and from those of 20 patients using (99m)Tc-ECD. We also measured BPI using GA. The BPI values obtained by SA (BPI(S)) (x) and by GA (BPI(G)) (y) correlated closely (y=0.708x+0.038, r=0.945 for (99m)Tc-HMPAO and y=0.559x+0.093, r=0.931 for (99m)Tc-ECD). However, the BPI(G) values were underestimated by 22.9%+/-6.6% (mean+/-SD) for (99m)Tc-HMPAO and by 27.9%+/-7.5% for (99m)Tc-ECD as compared with the BPI(S) values. The extent of underestimation tended to increase with increasing BPI(S) values. These findings were considered to be a result of the BPI(G) values being affected by the first-pass extraction fraction of the tracer. We also compared the BPI(S) and BPI(G) values with those of CBF measured using N-isopropyl-p-[(123)I]iodoamphetamine (CBF(IMP)) in 16 patients (six for (99m)Tc-HMPAO and ten for (99m)Tc-ECD). Although both BPI(S) and BPI(G) values correlated significantly with the CBF(IMP) values, the correlation coefficient in BPI(S) was always better than that in BPI(G) (r=0.869 for (99m)Tc-HMPAO and r=0.929 for (99m)Tc-ECD in BPI(S), r=0.629 for (99m)Tc-HMPAO and r=0.856 for (99m)Tc-ECD in BPI(G)). These results suggest that SA can provide a more reliable BPI for quantifying CBF using (99m)Tc-HMPAO or (99m)Tc-ECD than the conventional method using GA. Our method will be useful especially when using a tracer with a low first-pass extraction fraction and/or when performing activation studies using pharmacological intervention.

Aged↗

Pharmacokinetic analysis and antitumor efficacy of MKT-077, a novel antitumor agent.

MKT-077 (1-ethyl-2-[[3-ethyl-5-(3-methylbenzothiazolin-2-yliden)]-4- oxothiazolidin-2-ylidenemethyl] pyridinium chloride), a novel rhodacyanine dye in phase I/II clinical trials, may provide a new approach to cancer therapy based on the accumulation in the mitochondria of the cells of certain carcinomas, for example, those of the colon, breast and pancreas. To support the development of MKT-077 for clinical application as an intravenous (i.v.) therapy, we investigated the metabolic fate of [14C]MKT-077 in BDF1 mice as well as the distribution of MKT-077 in experimental LS174T tumor-bearing mice using a high-performance liquid chromatography (HPLC) method. The plasma levels of 14C after i.v. administration of [14C]MKT-077 declined in a triphasic manner. In the first distribution phase, the levels of 14C decreased with a T1/2 of approximately 5 min. In the second and terminal phase, the T1/2 of 14C was 2.8-4.6 h and 16.2 h, respectively. Cmax (1 min after injection) increased from 0.3 to 1.5 microg/ml linearly, but less than proportionately between the doses. The AUC(0-infinity) at 0.3, 1 and 3 mg/kg were 0.030 +/- 0.002, 0.60 +/- 0.12 and 1.73 +/- 0.25 microg x h/ml, respectively. Plasma clearance was approximately 1.8 l/h per kg (at doses of 1 and 3 mg/kg). The steady state volume of distribution (6.8 and 25.1 l/kg) indicated that MKT-077 distributed as a lipid-soluble molecule. The mean residence time (MRT) was 4.1 (at a dose of 1 mg/kg) and 14.1 h (at a dose of 3 mg/kg). In the first rapid phase (5 min after dosing), 14C radioactivity was detected in most of the tissues and organs, most strongly in the kidney cortex, and not in the central nervous system and testes. In the terminal phase (24 h after dosing), 14C contents increased in the intestinal tract, and in the kidney and liver were nearly to the background level. After i.v. bolus administration at a dose of 3 mg/kg of [14C]MKT-077, the predominant route of elimination of the radioactivity was via the feces, and recoveries of total radioactivity in urine and feces corresponded to 33.5% and 61.1%, respectively. More than 60% was recovered within 24 h and 95% within 1 week. MKT-077 was primarily excreted in unmetabolized form with five unidentified metabolites found in the urine and plasma. Intact MKT-077 was retained in the tumor tissue longer than in plasma and kidney in LS174T tumor-bearing mice receiving MKT-077 at an i.v. therapeutic dose (10 mg/kg). This accumulation decreased very slowly, suggesting that the high membrane potentials of tumor cell mitochondria may help retain the drug in tumors.

Animals↗

Amelioration by KRP-297, a new thiazolidinedione, of impaired glucose uptake in skeletal muscle from obese insulin-resistant animals.

We examined the effect of KRP-297, a new thiazolidinedione derivative, on glucose uptake in the soleus muscle of two animal models of insulin resistance that show moderate (ob/ob mice) and severe (db/db mice) hyperglycemia. Insulin-stimulated 2-deoxyglucose (2DG) uptake in soleus muscle was 53.8% lower in ob/ob mice versus lean mice (P < .05). When administered to ob/ob mice, KRP-297 (0.3 to 10 mg/kg) decreased plasma glucose and insulin levels and improved the impaired insulin-stimulated 2DG uptake in soleus muscle in a dose-dependent manner. Soleus muscle from db/db mice exhibited defects in both basal (35.0% decrease, P < .01) and insulin-stimulated (50.5% decrease, P < .01) 2DG uptake. These defects were improved by treatment with KRP-297 (0.3 to 10 mg/kg). Moreover, KRP-297 prevented severe hyperglycemia and the marked decrease in pancreatic insulin content in db/db mice. These results suggest that KRP-297 treatment is useful to prevent the development of diabetic syndromes in addition to ameliorating the impaired glucose transport in skeletal muscle.

Animals↗

Covalent structures of BmK AS and BmK AS-1, two novel bioactive polypeptides purified from Chinese scorpion Buthus martensi Karsch.

Complete amino acid sequences of two novel bioactive polypeptides, each containing 66 amino acid residues, BmK AS and BmK AS-1 purified from the venom of Chinese scorpion Buthus martensi Karsch, have been determined by Edman sequencing and mass spectrometry on native proteins, reduced and S-carboxymethylated proteins and their peptides obtained after cleavage with proteolytic enzymes. Sequence analysis showed 86.4% structural identity between BmK AS and BmK AS-1 and also a high sequence similarity between BmK ASs and AaH IT4, a unique anti-insect toxin and a ligand of Na+ channels obtained from Sahara scorpion A. australis Hector, but poor sequence homology between BmK ASs and those of the known alpha-, beta-type and long-chain insect-selective type scorpion neurotoxins. The positions of four disulfide bridges in BmK AS-1 were established as Cys-12 and Cys-62, Cys-16 and Cys-37, Cys-23 and Cys-44, and Cys-27 and Cys-46, which are the same as those in alpha- and beta-scorpion neurotoxins. These results suggest that BmK ASs and AaH IT4 may form a new group sharing similar structural and functional properties in the family of scorpion neurotoxic polypeptides.

Amino Acid Sequence↗

Dual interaction producing both territorial and schooling behavior in fish.

Fish schools are frequently observed without a leader and an explicit condition forming school. Several models of schooling have been proposed focusing on the influence of neighbors, and introducing probability distributions, while these models are based on the separation of schooling and territorial behavior. We frequently consider the duality of aggregation of animals, in which behavioral patterns involve both attraction and repulsion, antagonistic with each other. The idea of probability does not explain this duality that can provide both schooling and territorial behavior. From these biological facts, we have constructed a behavior model in which the influence of neighbors is formulated by the interface between the states of neighbors and a map of changes in these states. This interface uses a self-similar nowhere differentiable transition map which is temporally constructed, encompassing a crucial duality of repulsive and attractive forces hidden in the interaction among fishes. We tested it with computer simulations against the biological reality of schooling and territorial behavior. Due to the influence neighbors can have on duality, the same model can show both schooling behavior with a high degree of polarization and territorial behavior.

Animals↗

Clinical and laboratory features of lupus patients with complicating pulmonary disease.

The objective of this study was to determine the incidence of pulmonary involvement in patients with systemic lupus erythematosus (SLE) and to clarify the clinical and laboratory characteristics in SLE patients with various pulmonary involvements. A retrospective study (n = 137) revealed that the types of pulmonary involvement found in SLE patients were: pleuritis (9%), interstitial pneumonia (8%), pulmonary infarction (7%), pulmonary infection (4%), pulmonary hypertension (2%), restrictive dysfunction (28%) and decreased diffusion capacity (43%). The incidences of pericarditis (P < 0.01), arthralgia (P < 0.05) and hypoalbuminemia (P < 0.05) were significantly greater in patients with pleuritis than in those without, while in patients with interstitial pneumonia, the incidence of anti-SS-A antibody (P < 0.05) and sicca syndrome (P < 0.05) were significantly greater than in those without. A longitudinal follow-up study of patient groups with various pulmonary involvements revealed: 1. significant changes of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), lactate dehydrogenase (LDH) and thrombomodulin (TM) in patients with pleuritis, and 2. significant changes of WBC and LDH in patients with interstitial pneumonia. The increased ESR, CRP and TM levels during disease episodes suggest that the involvement of inflammatory processes is related to vasculitic events in the pathogenesis of lupus pleuritis. A higher incidence of anti-SS-A antibody in lupus patients with interstitial pneumonia suggests a potential role for this autoantibody in the pathogenesis of this complication.

Adolescent↗

Regional differences on production of chemokines in gastric mucosa between Helicobacter pylori-positive duodenal ulcer and gastric ulcer.

It is well known that antrum-predominant gastritis and pan-gastritis occurs in the patients with Helicobacter pylori-positive duodenal ulcer (DU) and gastric ulcer (GU), respectively. However, the role of chemokines in the pathogenesis of these pathologies is unclear. We examined the regional differences in mucosal chemokine production in patients with DU and GU. The production of interleukin-8 (IL-8), growth-related gene (GRO) alpha, and macrophage inflammatory protein (MIP)-1alpha was greater in the antrum than in the corpus in DU patients. In the patients with GU, monocyte chemoattractant protein (MCP)-1 levels in the mucosa adjacent to ulcer were greater than those away for the ulcer in the corpus. The reduction in chemokine production occurring in association with the eradication of H. pylori differed between DU and GU patients in the antrum (IL-8, P = 0.0394; GROalpha, P = 0.0149; MIP-1alpha, P = 0.0246; MCP-1, P = 0.0087). The data imply a different pathogenesis may exist for the gastritis present in patients with DU and GU occurring in H. pylori-positive individuals.

Adult↗

Experience with emergency cardiac surgery following institution of percutaneous cardiopulmonary support.

Between August 1992 and February 1998, 43 patients were treated with percutaneous cardiopulmonary support (PCPS) in our institution, and 8 of them subsequently required emergency cardiac surgery. There were 3 males and 5 females with a mean age of 63 years (range, 37 to 81 years). The etiology of shock in these 8 patients was acute myocardial infarction in 3, postinfarction left ventricular (LV) free wall rupture in 1, postinfarction ventricular septal perforation (VSP) in 1, LV free wall rupture and VSP in 1, and fatal arrhythmia due to severe aortic valvular disease in 2. The mean time interval from the onset of cardiogenic shock to the institution of PCPS was 77 min (range, 18 to 183 min). The mean time interval from the institution of PCPS until surgery was 145 min (range, 40 to 603 min). The surgical procedures were coronary artery bypass grafting (CABG) in 3 patients, closure of the LV rupture and/or closure of VSP in 3, and aortic valve replacement in 2. Six patients were weaned from PCPS, and 2 patients were discharged from the hospital (discharge rate, 25%). Although the results of emergency cardiac surgery following PCPS still are not satisfactory, we continue to apply PCPS and perform appropriate surgical procedures to improve the survival rate of the patients who would die without PCPS.

Adult↗

Kinetic parameter estimation from compartment models using a genetic algorithm.

Kinetic parameters were estimated from a three-compartment fluorodeoxyglucose model with three rate constants using a genetic algorithm. The performance of the genetic algorithm was investigated by simulation studies, in which brain time-activity data (TAD) were generated using cited mean values of rate constants and the plasma TAD obtained from positron emission tomographic studies. The accuracy of kinetic parameter estimation using the genetic algorithm was compared with that using the non-linear least-squares (NLSQ) method. The margin of error in the parameters estimated using the genetic algorithm tended to be smaller than that obtained by the NLSQ method. Although not statistically significant at a noise level of 5% in the brain TAD, the difference between the two methods became significant for all parameters at a noise level of 15% or higher. Our results suggest that the genetic algorithm is a promising means of estimating kinetic parameters from compartment models, because it is more robust against statistical noise than the NLSQ method and it can be rendered highly parallel for processing.

Algorithms↗

Hepatic extraction fraction of hepatobiliary radiopharmaceuticals measured using spectral analysis.

Measuring the hepatic extraction fraction (HEF) of a hepatobiliary radiopharmaceutical helps to differentiate hepatocyte from biliary tract diseases, and it is generally performed using deconvolution analysis. In this study, we measured HEF using spectral analysis. With spectral analysis, HEF was calculated from (the sum of the spectral data obtained by spectral analysis--the highest frequency component of the spectrum) divided by (the sum of the spectral data) x 100 (%). We applied this method to dynamic liver scintigraphic data obtained from six healthy volunteers and from 46 patients with various liver diseases, using 99Tcm-N-pyridoxyl-5-methyltryptophan (PMT). We also measured HEF using deconvolution analysis, in which the modified Fourier transform technique was employed. The HEF values obtained by spectral analysis correlated closely with those obtained by deconvolution analysis (r = 0.925), suggesting our method is valid. The HEF values obtained by spectral analysis decreased as the severity of liver disease progressed. The values were 100.0 +/- 0.0%, 94.7 +/- 13.6%, 76.2 +/- 27.4%, 45.7 +/- 15.6%, 82.7 +/- 24.2% and 95.2 +/- 11.8% (mean +/- S.D.) for the normal controls (n = 6), mild liver cirrhosis (n = 16), moderate liver cirrhosis (n = 11), severe liver cirrhosis (n = 5), acute hepatitis (n = 8) and chronic hepatitis groups (n = 6), respectively. The HEF was obtained more simply and rapidly by spectral analysis than by deconvolution analysis. The results suggest that our method using spectral analysis can be used as an alternative to the conventional procedure using deconvolution analysis for measuring HEF.

Aged↗

Identification of several clinical isolates of dermatophytes based on the nucleotide sequence of internal transcribed spacer 1 (ITS 1) in nuclear ribosomal DNA.

Nucleotide sequences of internal transcribed spacer 1 (ITS 1) in nuclear ribosomal DNA from seven morphologically unidentified dermatophyte isolates were determined. The sequences were compared with those of typical isolates of Trichophyton (T.) mentagrophytes var. interdigitale, T. rubrum, and Epidermophyton floccosum. Two of the isolates were classified as T. rubrum and the other five as T. mentagrophytes var. interdigitale. The results did not conflict with identifications using other molecular techniques, including random amplification of polymorphic DNA (RAPD) analysis and restriction enzyme analysis of mitochondrial DNAs. Thus, the nuclotide sequence of ITS 1 is possibly a good molecular marker for identification of these major anthropophilic dermatophyte species.

Base Sequence↗

Phylogenetic classification and species identification of dermatophyte strains based on DNA sequences of nuclear ribosomal internal transcribed spacer 1 regions.

The mutual phylogenetic relationships of dermatophytes of the genera Trichophyton, Microsporum, and Epidermophyton were demonstrated by using internal transcribed spacer 1 (ITS1) region ribosomal DNA sequences. Trichophyton spp. and Microsporum spp. form a cluster in the phylogenetic tree with Epidermophyton floccosum as an outgroup, and within this cluster, all Trichophyton spp. except Trichophyton terrestre form a nested cluster (100% bootstrap support). Members of dermatophytes in the cluster of Trichophyton spp. were classified into three groups with ITS1 homologies, with each of them being a monophyletic cluster (100% bootstrap support). The Arthroderma vanbreuseghemii-Arthroderma simii group consists of A. vanbreuseghemii, A. simii, Trichophyton mentagrophytes isolates from humans, T. mentagrophytes var. quinckeanum, Trichophyton tonsurans, and Trichophyton schoenleinii. Arthroderma benhamiae, T. mentagrophytes var. erinacei, and Trichophyton verrucosum are members of the Arthroderma benhamiae group. Trichophyton rubrum and Trichophyton violaceum form the T. rubrum group. This suggests that these "species" of dermatophytes have been overclassified. The ITS1 sequences of 11 clinical isolates were also determined to identify the species, and all strains were successfully identified by comparison of their base sequences with those in the ITS1 DNA sequence database.

Arthrodermataceae↗

Bacterial interdigital scaly erythema (Kitamura): a possible new clinical entity.

Four patients with scaly erythema on their finger webs and sides of their fingers during summer are described. These patients were working in either butcher's shops or a sushi bar, where they handled raw meat, chicken or fish for many hours. The eruptions first appeared as scaly erythema, sometimes accompanied by small pustules, on the second, third and fourth finger webs, and later the erythema extended to the sides of the fingers and palms. Maceration and/or shallow erosion sometimes appeared on the finger webs. Symptoms were usually mild; the patients complained of slight itching, irritation or pain. Direct examination of specimens stained with Parker blue-black ink containing KOH revealed scales containing bacterial granules or filaments. Several species of bacteria were cultured including Corynebacterium sp. Fungus was not detected in either KOH specimens or in cultures. The lesions responded rapidly to topical or oral antibiotics; however, they recurred frequently during hot and humid weather. Hitherto a similar condition has not been described and is possibly a new clinical entity.

Adolescent↗

Glicentin and oxyntomodulin modulate both the phosphoinositide and cyclic adenosine monophosphate signaling pathways in gastric myocytes.

We have investigated the transduction pathways mediating the contractile effect of two glucagon-containing peptides, glicentin (GLIC) and oxyntomodulin (OXM), on smooth muscle cells isolated from rabbit antrum. Low concentrations of GLIC induced a biphasic and rapid (first phase at 5-8 sec) Ins(1,4,5)P3 production. By comparison, higher concentrations of OXM or OXM(19-37) were required to obtain biphasic time-courses of Ins(1,4,5)P3 production. In a Ca2+ free medium, the first phase of Ins(1,4,5)P3 production induced by GLIC or OXM was maintained, while the second phase disappeared. In saponin-permeabilized cells, all three peptides induced cell contraction with similar efficacies and potencies. Exogenous Ins(1,4,5)P3 mimicked the contractile effect of the peptides and heparin, which inhibits the Ins(1,4,5)P3 binding to its receptor, prevented contraction stimulated by each effector. We conclude that a Ca2+ mobilization from the intracellular stores is essential in the contractile effects of GLIC and OXM. Using the fluo-3 probe, a [Ca2+]i increase was observed in the presence of GLIC, OXM, or OXM(19-37). The three peptides reduced by 30-40% the cAMP content of cells stimulated by forskolin. This effect was pertussis toxin sensitive as demonstrated with OXM(19-37). Our data constitute important clues for the existence in smooth muscle cells of receptor(s) specific for the GLIC/OXM hormones, coupled via G protein(s) to both Ca2+ and cAMP pathways.

Animals↗

Histamine release induced by immobilization, gentle handling and decapitation from mast cells and its inhibition by nedocromil in rats.

The effect of immobilization, gentle handling and decapitation on the level of plasma histamine in Wistar rats was investigated. Mast cell deficient (Ws/Ws) rats were used to characterize the source of elevated histamine in plasma by stress, and the effect of nedocromil, a mast cell stabilizer, on histamine release was assessed in these models in vivo. The plasma histamine concentration of freely moving rats was 93.0+/-2.3 pmol/ml. Gentle handling produced a transient increase in plasma histamine level by 1.9-fold, whereas immobilization resulted in a longer-lasting elevation by 2.6-fold compared to that in the freely moving rats. Decapitation increased the plasma histamine level by 10- to 16-fold compared with that in the freely moving rats. No increase in plasma histamine was found in Ws/Ws rats exposed to stress. Nedocromil inhibited the increase in plasma histamine level induced by stress in a dose-dependent manner. These findings suggest that stress induces histamine release from mast cells in Wistar rats and the extent of this histamine release increases with the severity of stress. Nedocromil proved to be a good pharmacological tool to inhibit stress-induced release of mediators from mast cells.

Animals↗