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Biomedical subjects

Tetsuo Hashimoto

Publications and source records attributed to Tetsuo Hashimoto.

At least 19 recordsLinked to original sources

Usability of VTL from natural quartz grains for retrospective dosimetry.

To develop retrospective dosimetry of unexpected radiation accident, basic studies on violet thermoluminescence (VTL) phenomena were conducted using natural quartz grains. All VTL glowcurves of as-received samples did not exhibit peaks <250 degrees C, although for artificially irradiated quartz samples there were VTL peaks in the temperature region <250 degrees C. Therefore, accident doses could be estimated without the interference of naturally accumulated doses by VTL measurements from natural quartz. The mean lives of VTL were evaluated by the various heating rates method and the range of values was found to be between some days and ten thousands of years depending on each peak. Especially, the mean life of VTL peak at 200 degrees C was years order. Furthermore, the lower detection limit was calculated to be tens of mGy from the response curve. This value was lower than that of other methods such as ESR dosimetry. From these results, we conclude that VTL dosimetry can be preferred for accidental evaluation.

Electrons↗

[A case of takotsubo cardiomyopathy provoked by taking a new quinolone antibiotic drug and a non-steroidal anti-inflammatory drug].

A 97-year-old woman was provoked a convulsion after taking a new quinolone antibiotic drug Levofloxacin and a non-steroidal anti-inflammatory drug Loxoprofen. At a later time, she was suffered from severe chest pain. An electrocardiogram showed ST segment elevation in leads II, III and aVF, and T-wave inversion in leads V1 to V4. Coronary angiography demonstrated no organic stenosis, however biventriculography revealed apical ballooning akinesis and basal hyperkinesis. Positron emission tomography was also performed to assess the uptake of 18F-fluorodeoxyglucose (FDG) after 75 g oral glucose loading for evaluating myocardial glucose metabolism at 10th day. Severely reduced uptake of FDG was observed in the apical ballooning region. Left ventriculography showed normal wall motion at 19th day. Thallium-201 myocardial single-photon emission computed tomography (SPECT) to determine the status of myocardial perfusion at the 20th hospital day showed normal perfusion. Iodine-123-beta-methyl-p-iodophenyl penta-decanoic acid myocardial SPECT to evaluate myocardial fatty acid metabolism at the 23rd day revealed severely reduced uptake in the apical ballooning region. These findings suggested that the coronary microcirculation was impaired in the apical ballooning region.

Aged, 80 and over↗

[Clinical usefulness of 1231I-BMIPP myocardial SPECT in patient with microvascular spasm: a case report].

This patient was a 64-year-old man with chest pain at rest. An electrocardiogram showed depression of the ST segment in V2-V5 leads during chest pain. 123I-BMIPP myocardial SPECT revealed reduced uptake in the apex. Coronary angiographies revealed severely delayed filling of contrast medium without narrowing of epicardial coronary arteries. An intracoronary infusion of isosorbide dinitrate did not improve the delayed filling of contrast medium or the ST segment depression. After an intracoronary infusion of nicorandil, coronary arterial flows were remarkably improved, chest symptoms disappeared, and electrocardiographic findings were improved. Left ventriculography showed severe hypokinesis in the apex. After the medication with nicorandil, reduced 123I-BMIPP myocardial uptake and reduced wall motion on echocardiography were improved. These findings suggest that myocardial ischemia in this case might be explained as having been caused by microcirculation disturbance.

Coronary Circulation↗

[Clinical usefulness of 201Tl/99mTc-PYP dual myocardial quantitative gated SPECT program using low-dose dobutamine loading in assessment of myocardial viability in patient with acute myocardial infarction--a case report].

An 86-year-old man with chest pain was admitted to our hospital. Coronary angiography revealed 99% stenosis of the mid segment of the left anterior descending coronary artery, therefore, a coronary stent was implanted. Immediately after the stent implantation, 99% stenosis occurred at the proximal site of the 1st diagonal artery because of stent jeal. On the 4th hospital day, ECG-gated 201TL/99mTc-PYP dual myocardial quantitative gated SPECT was performed at rest and during low-dose dobutamine loading. The 201Tl scintigraphy revealed moderately reduced uptake in the anterior, septal and apical walls, and 99mTc-PYP uptake was observed in the mid-anterior wall. A three-dimensional surface display of gated 201Tl SPECT images showed severe hypokinesis in the anterior, septal and apical walls at rest. On the other hand, during low-dose dobutamine loading, improved wall motion was observed in the basal anterior and septal walls, while no change was observed in the midanterior and apical wall movements. Three-dimensional surface display of gated 201Tl/99mTc-PYP dual SPECT images revealed similar patterns of wall motion as those of gated 201Tl SPECT images at rest. During low-dose dobutamine loading, on the other hand, a three-dimensional surface display of gated 201Tl/99mTc-PYP dual SPECT images revealed improved wall motion in the basal anterior, septal and apical walls, but worsened wall motion of the mid-anterior wall. After 6 months, a follow-up coronary angiography revealed no re-stenosis of the stent, but 99% stenosis at the proximal aspect of the 1st diagonal artery. Left ventriculography revealed improved wall motion in the apex and akinesis of the mid-anterior wall. These wall motion findings were similar to those visualized in the three-dimensional surface display of gated 201Tl/99mTc-PYP dual SPECT images during low-dose dobutamine loading in the acute phase. These results suggest that 201Tl/99mTc-PYP dual myocardial quantitative gated SPECT using low-dose dobutamine loading could be useful for the assessment of myocardial viability after reperfusion therapy in patients with acute myocardial infarction.

Aged, 80 and over↗

Phylogeny of the Centrohelida inferred from SSU rRNA, tubulins, and actin genes.

Amoeboid protists are major targets of recent molecular phylogeny in connection with reconstruction of global phylogeny of eukaryotes as well as the search for the root of eukaryotes. The Centrohelida are one of the major groups of Heliozoa, classified in the Actinopodida, whose evolutionary position is not well understood. To clarify the relationships between the Centrohelida and other eukaryotes, we sequenced SSU rRNA, alpha-tubulin, and beta-tubulin genes from a centroheliozoan protist, Raphidiophrys contractilis. The SSU rRNA phylogeny showed that the Centrohelida are not closely related to other heliozoan groups, Actinophryida, Desmothoracida, or Taxopodida. Maximum likelihood analyses of the combined phylogeny using a concatenate model for an alpha- + beta-tubulin + actin data set, and a separate model for SSU rRNA, alpha- and beta-tubulin, and actin gene data sets revealed the best tree, in which the Centrohelida have a closer relationship to Rhodophyta than to other major eukaryotic groups. However, both weighted Shimodaira-Hasegawa and approximately unbiased tests for the concatenate protein phylogeny did not reject alternative trees in which Centrohelida were constrained to be sisters to the Amoebozoa. Moreover, alternative trees in which Centrohelida were placed at the node branching before and after Amoebozoa or Viridiplantae were not rejected by the WSH tests. These results narrowed the possibilities for the position of Centrohelida to a sister to the Rhodophyta, to the Amoebozoa, or to an independent branch between the branchings of Amoebozoa and Rhodophyta (or possibly Plantae) at the basal position within the bikonts clade in the eukaryotic tree.

Actins↗

The origin of dihydroorotate dehydrogenase genes of kinetoplastids, with special reference to their biological significance and adaptation to anaerobic, parasitic conditions.

Trypanosoma cruzi dihydroorotate dehydrogenase (DHOD), the fourth enzyme of the de novo pyrimidine biosynthetic pathway, is localized in the cytosol and utilizes fumarate as electron acceptor (fumarate reductase activity), while the enzyme from other various eukaryotes is mitochondrial membrane-linked. Here we report that DHOD-knockout T. cruzi did not express the enzyme protein and could not survive even in the presence of pyrimidine nucleosides, substrates for the potentially active salvage pathway, suggesting a vital role of fumarate reductase activity in the regulation of cellular redox balance. Cloning and phylogenetic analysis of euglenozoan DHOD genes showed that the euglenoid Euglena gracilis had a mitochondrial DHOD and that biflagellated bodonids, a sister group of trypanosomatids within kinetoplastids, harbor the cytosolic DHOD. Further, Bodo saliens, a bodonid, had an ACT/DHOD gene fusion encoding aspartate carbamoyltransferase (ACT), the second enzyme of the de novo pyrimidine pathway, and DHOD. This is the first report of this novel gene structure. These results are consistent with suggestions that an ancient common ancestor of Euglenozoa had a mitochondrial DHOD whose descendant exists in E. gracilis and that a common ancestor of kinetoplastids (bodonids and trypanosomatids) subsequently acquired a cytosolic DHOD by horizontal gene transfer. The cytosolic DHOD gene thus acquired may have contributed to adaptation to anaerobiosis in the kinetoplastid lineage and further contributed to the subsequent establishment of parasitism in a trypanosomatid ancestor. Different molecular strategies for anaerobic adaptation in pyrimidine biosynthesis, used by kinetoplastids and by euglenoids, are discussed. Evolutionary implications of the ACT/DHOD gene fusion are also discussed.

Adaptation, Physiological↗

[Assessment of microcirculation disturbance in patients with coronary ectasia by ATP-loading 99mTc-tetrofosmin myocardial SPECT].

UNLABELLED: Patients with coronary ectasia often develop chest pain and reveal ischemic changes on electrocardiograms and reduced left ventricular wall motion on left ventriculography, in the absence of epicardial coronary artery stenotic regions. We examined the disturbances in the coronary microcirculation in patients with coronary ectasia using left ventriculography and ATP loading 99mTc-tetrofosmin myocardial single photon emission computed tomography (SPECT) before and after administration of a coronary vasodilator and antiplatelet agents. METHODS: Twenty patients in whom coronary angiography revealed diffuse coronary artery ectasia but no stenotic regions were enrolled in this study. Left ventriculography and ATP loading 99mTc-tetrofosmin myocardial SPECT were performed before and after administration of the coronary vasodilator, nicorandil, as well as that of the antiplatelet agents, aspirin and ticlopidine. RESULTS: (1) The ejection fraction in left ventriculography was 48.3 +/- 17.4% before, and 56.6 +/- 18.3% after the drug administration, the ejection fraction was improved after the drug administration (p < 0.05). (2) Before the drug administration, the total defect scores on 99mTc-tetrofosmin myocardial SPECT were 5.9 +/- 3.1 and 8.8 +/- 2.7 in the ATP-loading and rest images, respectively (p < 0.05), and the corresponding scores after the drug administration were 4.1 +/- 3.0 and 5.4 +/- 3.1, respectively (N.S.). Thus, the total defect scores in the ATP-loading and rest images improved after the drug administration (p < 0.05). CONCLUSION: Myocardial damage in patients with coronary ectasia might be induced by microthrombotic embolism and microcirculation disturbance.

Adenosine Triphosphate↗

Root of the Eukaryota tree as inferred from combined maximum likelihood analyses of multiple molecular sequence data.

Extensive studies aiming to establish the structure and root of the Eukaryota tree by phylogenetic analyses of molecular sequences have thus far not resulted in a generally accepted tree. To re-examine the eukaryotic phylogeny using alternative genes, and to obtain a more robust inference for the root of the tree as well as the relationship among major eukaryotic groups, we sequenced the genes encoding isoleucyl-tRNA and valyl-tRNA synthetases, cytosolic-type heat shock protein 90, and the largest subunit of RNA polymerase II from several protists. Combined maximum likelihood analyses of 22 protein-coding genes including the above four genes clearly demonstrated that Diplomonadida and Parabasala shared a common ancestor in the rooted tree of Eukaryota, but only when the fast-evolving sites were excluded from the original data sets. The combined analyses, together with recent findings on the distribution of a fused dihydrofolate reductase-thymidylate synthetase gene, narrowed the possible position of the root of the Eukaryota tree on the branch leading to Opisthokonta or to the common ancestor of Diplomonadida/Parabasala. However, the analyses did not agree with the position of the root located on the common ancestor of Opisthokonta and Amoebozoa, which was argued by Stechmann and Cavalier-Smith [Curr. Biol. 13:R665-666, 2003] based on the presence or absence of a three-gene fusion of the pyrimidine biosynthetic pathway: carbamoyl-phosphate synthetase II, dihydroorotase, and aspartate carbamoyltransferase. The presence of the three-gene fusion recently found in the Cyanidioschyzon merolae (Rhodophyta) genome sequence data supported our analyses against the Stechmann and Cavalier-Smith-rooting in 2003.

Animals↗

Simultaneous determination of radionuclides separable into natural decay series by use of time-interval analysis.

A delayed coincidence method, time-interval analysis (TIA), has been applied to successive alpha- alpha decay events on the millisecond time-scale. Such decay events are part of the (220)Rn-->(216)Po ( T(1/2) 145 ms) (Th-series) and (219)Rn-->(215)Po ( T(1/2) 1.78 ms) (Ac-series). By using TIA in addition to measurement of (226)Ra (U-series) from alpha-spectrometry by liquid scintillation counting (LSC), two natural decay series could be identified and separated. The TIA detection efficiency was improved by using the pulse-shape discrimination technique (PSD) to reject beta-pulses, by solvent extraction of Ra combined with simple chemical separation, and by purging the scintillation solution with dry N(2) gas. The U- and Th-series together with the Ac-series were determined, respectively, from alpha spectra and TIA carried out immediately after Ra-extraction. Using the (221)Fr-->(217)At ( T(1/2) 32.3 ms) decay process as a tracer, overall yields were estimated from application of TIA to the (225)Ra (Np-decay series) at the time of maximum growth. The present method has proven useful for simultaneous determination of three radioactive decay series in environmental samples.

Journal Article↗

Direct evidence for cyanide-insensitive quinol oxidase (alternative oxidase) in apicomplexan parasite Cryptosporidium parvum: phylogenetic and therapeutic implications.

Cryptosporidium parvum is a parasitic protozoan that causes the diarrheal disease cryptosporidiosis, for which no satisfactory chemotherapy is currently available. Although the presence of mitochondria in this parasite has been suggested, its respiratory system is poorly understood due to difficulties in performing biochemical analyses. In order to better understand the respiratory chain of C. parvum, we surveyed its genomic DNA database in GenBank and identified a partial sequence encoding cyanide-insensitive alternative oxidase (AOX). Based on this sequence, we cloned C. parvum AOX (CpAOX) cDNA from the phylum apicomplexa for the first time. The deduced amino acid sequence (335 a.a.) of CpAOX contains diiron coordination motifs (-E-, -EXXH-) that are conserved among AOXs. Phylogenetic analysis suggested that CpAOX is a mitochondrial-type AOX, possibly derived from mitochondrial endosymbiont gene transfer. The recombinant enzyme expressed in Escherichia coli showed quinol oxidase activity. This activity was insensitive to cyanide and highly sensitive to ascofuranone, a specific inhibitor of trypanosome AOX.

Amino Acid Sequence↗

Comparative analysis of the ribosomal components of the hydrogenosome-containing protist, Trichomonas vaginalis.

The ribosomes of the amitochondriate but hydrogenosome-containing protist lineage, the trichomonads, have previously been reported to be prokaryotic or primitive eukaryotic, based on evidence that they have a 70S sedimentation coefficient and a small number of proteins, similar to prokaryotic ribosomes. In order to determine whether the components of the trichomonad ribosome indeed differ from those of typical eukaryotic ribosomes, the ribosome of a representative trichomonad, Trichomonas vaginalis, was characterized. The sedimentation coefficient of the T. vaginalis ribosome was smaller than that of Saccharomyces cerevisiae and larger than that of Escherichia coli. Based on two-dimensional PAGE analysis, the number of different ribosomal proteins was estimated to be approximately 80. This number is the same as those obtained for typical eukaryotes (approximately 80) but larger than that of E. coli (approximately 55). N-Terminal amino acid sequencing of 18 protein spots and the complete sequences of 4 ribosomal proteins as deduced from their genes revealed these sequences to display typical eukaryotic features. Phylogenetic analyses of the five ribosomal proteins currently available also clearly confirmed that the T. vaginalis sequences are positioned within a eukaryotic clade. Comparison of deduced secondary structure models of the small and large subunit rRNAs of T. vaginalis with those of other eukaryotes revealed that all helices commonly found in typical eukaryotes are present and conserved in T. vaginalis, while variable regions are shortened or lost. These lines of evidence demonstrate that the T. vaginalis ribosome has no prokaryotic or primitive eukaryotic features but is clearly a typical eukaryotic type.

Amino Acid Sequence↗

Molecular cloning and characterization of Trypanosoma vivax alternative oxidase (AOX) gene, a target of the trypanocide ascofuranone.

Trypanosoma vivax causes nagana disease in cattle. Since T. vivax is transmitted not only by tsetse flies but also by other biting flies (non-cyclic transmission), the parasite has been distributed to and has had a significant economic impact on wide geographical areas, including Africa and South America. Our previous study on Trypanosoma brucei brucei showed that the trypanosome alternative oxidase (TAO, TbAOX) is a promising target of chemotherapy. For this reason, we also have cloned the T vivax AOX (TvAOX) gene and characterized the recombinant enzyme. The deduced amino acid sequence (328 a.a.) of TvAOX shares 76% identity with TbAOX and contains the diiron-coordination motifs (-E-, -EXXH-) that are conserved among AOXs. The Km of recombinant TvAOX (rTvAOX) expressed in Escherichia coli for ubiquinol (87.0 +/- 0.54 microM) was significantly lower than the value for recombinant TbAOX (rTbAOX) (714 +/- 4.5 microM). Ascofuranone, the most potent inhibitor of TbAOX, was a competitive inhibitor of rTvAOX with a Ki value (0.40 +/- 0.00 nM) significantly lower than that for rTbAOX (1.29 +/- 0.00 nM). The non-cyclic transmission ability of T. vivax and the in vivo chemotherapeutic efficacy of ascofuranone against T. vivax and T. b. brucei infection are discussed in terms of these Km and Ki values.

Amino Acid Sequence↗

Molecular and biochemical characterization of D-phosphoglycerate dehydrogenase from Entamoeba histolytica. A unique enteric protozoan parasite that possesses both phosphorylated and nonphosphorylated serine metabolic pathways.

A putative phosphoglycerate dehydrogenase (PGDH), which catalyzes the oxidation of d-phosphoglycerate to 3-phosphohydroxypyruvate in the so-called phosphorylated serine metabolic pathway, from the enteric protozoan parasite Entamoeba histolytica was characterized. The E. histolytica PGDH gene (EhPGDH) encodes a protein of 299 amino acids with a calculated molecular mass of 33.5 kDa and an isoelectric point of 8.11. EhPGDH showed high homology to PGDH from bacteroides and another enteric protozoan ciliate, Entodinium caudatum. EhPGDH lacks both the carboxyl-terminal serine binding domain and the 13-14 amino acid regions containing the conserved Trp139 (of Escherichia coli PGDH) in the nucleotide binding domain shown to be crucial for tetramerization, which are present in other organisms including higher eukaryotes. EhPGDH catalyzed reduction of phosphohydroxypyruvate to phosphoglycerate utilizing NADH and, less efficiently, NADPH; EhPGDH did not utilize 2-oxoglutarate. Kinetic parameters of EhPGDH were similar to those of mammalian PGDH, for example the preference of NADH cofactor, substrate specificities and salt-reversible substrate inhibition. In contrast to PGDH from bacteria, plants and mammals, the EhPGDH protein is present as a homodimer as demonstrated by gel filtration chromatography. The E. histolytica lysate contained PGDH activity of 26 nmol NADH utilized per min per mg of lysate protein in the reverse direction, which consisted 0.2-0.4% of a total soluble protein. Altogether, this parasite represents a unique unicellular protist that possesses both phosphorylated and nonphosphorylated serine metabolic pathways, reinforcing the biological importance of serine metabolism in this organism. Amino acid sequence comparison and phylogenetic analysis of various PGDH sequences showed that E. histolytica forms a highly supported monophyletic group with another enteric protozoa, cilliate E. caudatum, and bacteroides.

Amino Acid Sequence↗

Case of leiomyosarcoma of the renal pelvis.

A 54-year-old man presented at the Mibayashi clinic with bowel discomfort. Ultrasonography showed a left renal mass and the patient was referred to Noto General Hospital for urological evaluation. Results of the physical examination were unremarkable, but computed tomography and magnetic resonance imaging demonstrated a large tumor in the middle of the left kidney. The patient underwent left nephrectomy. On hemisection of the kidney, a firm tumor, measuring 8 x 7.5 cm, was seen occupying the renal pelvis. The histological diagnosis was leiomyosarcoma arising from the right renal pelvis. No treatment was provided after surgery and no recurrence was observed 6 months postoperatively.

Biopsy, Needle↗