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Biomedical subjects

H Takechi

Publications and source records attributed to H Takechi.

At least 19 recordsLinked to original sources
PubMed2001

Synthesis and evaluation of a difluoromethylene analogue of sphingomyelin as an inhibitor of sphingomyelinase.

A sphingomyelin analogue 2, in which the long alkenyl chain and the phosphodiester moiety of sphingomyelin were replaced by a phenyl and an isosteric difluoromethylenephosphonic acid, was prepared to evaluate its inhibitory potency to sphingomyelinase. The analogue non-competitively inhibited the neutral sphingomyelinase in bovine brain microsomes with an IC50 of 400 microM. The compound had the ability to suppress tumor necrosis factor alpha-induced apoptosis of PC-12 neurons at a low concentration of 0.1 microM.

Animals↗
PubMed2001

Cortical networks recruited for time perception: a monkey positron emission tomography (PET) study.

The presence of an "internal clock" in the brain has been assumed to underlie the information processing related to time. This clock plays a critical role in time keeping and time perception, which are closely associated with integrated functions in the brain. To identify the brain areas recruited for time keeping and time perception, we performed positron emission tomography (PET) studies with rhesus monkeys to measure regional cerebral blood flow (rCBF) as an index of neural activity during time discrimination tasks of different durations ranging from 400 to 1500 ms. Changes in rCBF that covaried significantly with the durations of the target being perceived by subjects were found in the dorsolateral prefrontal cortex (DLPFC), the posterior part of the inferior parietal cortex, basal ganglia, and posterior cingulate cortex. Furthermore, a loss of neuronal function in the DLPFC caused by a local application of bicuculline resulted in the selective reduction of performance in time discrimination tasks. The results indicate that a neural network composed of the posterior inferior parietal cortex to the DLPFC plays a crucial role in the temporal monitoring process in time perception.

Animals↗
PubMed2000

Screening search for organic fluorophores: syntheses and fluorescence properties of 3-azolyl-7-diethylaminocoumarin derivatives.

In order to find a highly sensitive fluorophore, 3-azolyl-7-diethylaminocoumarin derivatives were synthesized. Both the absorption and fluorescence maxima of the coumarin-thiazole compounds showed red shifts with increases of the molar absorptivities and fluorescence intensities, in comparison with those of the corresponding coumarin-oxazole compounds. Among them, 3-(5-ethoxycarbonyl-1,3-thiazol-2-yl)-7-diethylamino-2H-chromen -2-one (3e) was one of the most promising candidates as a fluorophore accessible for analytical purposes in the fields of analytical and biological chemistry.

Azoles↗
PubMed1999

A novel subtype of prostacyclin receptor in the central nervous system.

Recently, in the course of our search for the prostacyclin receptor in the brain, we found a novel subtype, designated as IP2, which was finely discriminated by use of the specific ligand (15R)-16-m-tolyl-17,18,19,20-tetranorisocarbacyclin (15R-TIC) and specifically localized in the rostral part of the brain. In the present study, the tritiated compound 15R-[15-(3)H]TIC was synthesized and utilized for more specific research on IP2. The specificity of binding to rat brain regions was confirmed by use of several prostacyclin derivatives including 15S-TIC. Mapping of 15R- and 15S-[3H]TIC binding in adjacent pairs of frozen sections of rat brain demonstrated a quite similar pattern of distribution in almost all rostral brain regions, indicating that the regions may contain only the IP2 subtype. On the other hand, 15R-[3H]TIC binding was very faint as compared with 15S-[3H]TIC binding in the caudal medullary region. High densities of 15R-[3H]TIC binding sites were shown in the dorsal part of the lateral septal nucleus, thalamic nuclei, limbic structures, and some of the cortical regions. Scatchard plot analysis showed two components of high-affinity 15R-[3H]TIC binding in the rostral regions, one with a K(D) value at approximately 1 nM and the other with approximately 30 nM. These results strengthen our previous finding that a different subtype of prostacyclin receptor is expressed in the CNS, and the map with 15R-[3H]TIC obtained here could guide further studies on the molecular and functional properties of the IP2.

Animals↗
PubMed1998

Evaluation of wheelchair cushions by means of pressure distribution mapping.

We studied the seated buttock pressure distribution in six paraplegic patients by means of computerized pressure mapping. They were all male and their age ranged from 18 to 48 years old. Their level of paralysis varied from Th5 to L1. Five kinds of wheelchair cushions were studied: an air cushion, a contour cushion, a polyurethane foam cushion, a Cubicushion (which is made of polyurethane foams) and a silicone gel cushion. A tactile sensor consisting of 2064 matrices was used for measuring the buttock pressure distribution and the data was analyzed on a personal computer. Peak pressures measured for each cushion were as follows (in descending order): the Cubicushion, the polyurethane foam cushion, the contour cushion, the silicone gel cushion, and the air cushion. The areas of total contact measured for each cushion were as follows (in descending order): the air cushion, the silicone gel cushion, the polyurethane foam cushion, the contour cushion and the Cubicushion. Based on these findings, we conclude that the most advantageous cushion is the air cushion or the silicone gel cushion. Likewise, we conclude that the Cubicushion is not practical for pressure sore prevention.

Adolescent↗
PubMed1997

Mapping of cortical areas involved in color vision in non-human primates.

Positron emission tomography (PET) was used to measure changes in the regional cerebral blood flow (rCBF) of rhesus monkeys performing visual discrimination tasks. In comparison with both position and brightness discrimination tasks, the color discrimination task activated the posterior inferior temporal cortex and a ventromedial occipital region, which is located along the anterior one-third of the calcarine sulcus. In contrast, the position task activated the middle temporal area and intraparietal cortex as compared with the color task. These results confirm the segregation of visual pathways and delineate the visual areas involved in color vision. This approach might bridge the gap between invasive studies in animals and functional imaging studies in humans.

Animals↗
PubMed1997

Analysis of stereoisomeric C27-bile acids by high performance liquid chromatography with fluorescence detection.

A method for differentially measuring the 24-hydroxylated stereoisomeric intermediates (3 alpha,7 alpha,12 alpha,24-tetrahydroxy- and 3 alpha,7 alpha,24-trihydroxy-5 beta-cholestan-26-oic acids) and related C27-bile acids in beta-oxidation of bile acid biosynthesis has been developed by high performance liquid chromatography with fluorescence detection. The method involved the derivatization of the above intermediable C27-bile acids into fluorescent esters with 3-(4-bromomethylphenyl)-7-diethylaminocoumarin, a newly synthesized labeling reagent for carboxylic acids. The fluorescent derivatives were subjected to a short silica gel column to eliminate interfering products prior to analysis by high performance liquid chromatography. The separation of the 16 kinds of bile acids containing stereoisomers was carried out using a reversed-phase Inertsil C8-column by gradient elution and detected with a fluorometer (Ex. 400 nm, Em. 475 nm). The linearity of calibration curve for each bile acid was from 0.5 to 250 pmol (r = 0.999) and the detection limits were about 15 fmol at a signal-to-noise ratio of 3. The method was applied to the determination of intermediates in beta-oxidation of bile acid biosynthesis using rat liver homogenate. The results showed that two stereoisomers of 24-hydroxylated C27-bile acids were predominantly produced, indicating the formation of the isomers by the cis-hydration with water.

Animals↗
PubMed1997

A survey of wheelchair use by paraplegic individuals in Japan. Part 1: Characteristics of wheelchair cushions.

The characteristics of wheelchair cushions used by 218 paraplegic patients who lived independent lives were surveyed to clarify the present state of wheelchair cushioning for pressure sore prevention in Japan. Out of 586 cushions surveyed, 91.0% were ready-made and the rest were custom-made. The outstanding popularity of polyurethane foam ready-made cushions (76.3%) suggested that insufficient consideration was taken in the selection of cushions. Custom-made cushions displayed unique modifications to relieve contact pressure or to stabilize sitting posture, which should be systematically provided for all patients. The variety of cushion types and the frequent dissatisfaction with cushions seen in patients with current pressure sores suggested a strong demand for the effective prescription of cushions. Furthermore, 30% of all cushions had had an excessively prolonged use, indicating insufficient follow-up. A medical system including deliberate prescription and regular follow-up of wheelchair cushions should be established for the effective prevention of pressure sores.

Adolescent↗
PubMed1997

A survey of wheelchair use by paraplegic individuals in Japan. Part 2: Prevalence of pressure sores.

A cross-sectional survey was done to clarify the incidence of pressure sores in 218 self-supported Japanese paraplegic patients and to determine effective measures for prevention. The majority of patients (85.7%) had previous pressure sores, and 46.3% had undergone multiple surgeries. Some patients (17.9%) were still suffering from persistent sores which commonly developed at the ischial tuberosities, suggesting insufficiency of self-care practice during wheelchair activities. Sensory disturbance over the seating surface, urinary incontinence, and general complications were seen in 85.8%, 49.5%, and 18.8% of total subjects, respectively. They were seen as risk factors for pressure sores, but only urinary incontinence clearly increased the current pressure sore prevalence. Nevertheless, both self-care practice and sports activities, seen in 85.3% and 36.2% of total subjects, respectively, contributed to greatly reduce the incidence. A patient education system including acquisition of basic knowledge and proper technique should be established to promote effective prevention of pressure sores in Japan.

Adolescent↗
PubMed1997

Survey of above knee (A/K) prostheses currently used in the Chugoku-Shikoku district of Japan.

To determine the extent to which recent advances in biomechanical technology have been implemented and to evaluate these new technologies, 84 unilateral above knee (A/K) amputees and their prostheses were surveyed in the Chugoku-Shikoku district of Japan, especially in regard to the types, sockets and components of A/K prostheses currently in use. Background factors such as age and sex of the A/K amputees and the period after amputation were also surveyed. Of the 84 amputees surveyed, 74 (88.1%) were over 40 years old and 40 (47.6%) were over 60 years old. There were 10 women (11.9%) and 74 men (88.1%). The period after amputation was under 25 years in 58 (69.0%) cases. Regarding the type of A/K prostheses, one-third of the prostheses was of the exoskeletal type and two-thirds were of the endoskeletal type. Although the endoskeletal type is becoming more popular recently, elderly A/K amputees tend to use the exoskeletal type. Thirty-one (36.9%) had plug-fit sockets which are preferable for those who follow the Japanese practice of sitting on the floor, especially for elderly amputees. Thirty-seven (44.0%) had a lock-knee, 27 (73.0%) of which were used by amputees over 60 years old. Seventy-three (86.9%) had a single-axis ankle which is generally considered to be the most stable ankle. Thus, the most common combination of prosthetic components for elderly A/K amputees was the plug-fit socket, lock-knee joint and single-axis ankle.

Adult↗
PubMed1996

A novel subtype of the prostacyclin receptor expressed in the central nervous system.

By use of several prostacyclin analogs and an in vitro autoradiographic technique, we have found a novel subtype of the prostacyclin receptor, one having different binding properties compared with those of the known prostacyclin receptor in the rat brain. Isocarbacyclin, which is a potent agonist for the known prostacyclin receptor, had high affinity for the novel subtype (dissociation constant (Kd) of 7.8 nM). However, iloprost, which is usually used as a stable prostacyclin analog, showed low affinity binding (Kd = 159 nM) for the subtype. Other prostaglandins showed no or little affinity for the subtype. [3H]Isocarbacyclin binding was high in the thalamus, lateral septal nucleus, hippocampus, cerebral cortex, striatum, and dorsal cochlear nucleus. Although the nucleus of the solitary tract and the spinal trigeminal nucleus showed a high density of [3H]isocarbacyclin binding, [3H]iloprost also had high affinity in these regions, and the binding specificity was similar to that for the known prostacyclin receptor. Hemilesion studies of striatal neurons lesioned by kainate or of dopaminergic afferents lesioned by 6-hydroxydopamine revealed that the binding sites of the novel subtype exist on neuronal cells in the striatum, but not on the presynaptic terminal of afferents or on glial cells. Electrophysiological studies carried out in the CA1 region of the hippocampus revealed that prostacyclin analogs have a facilitatory effect on the excitatory transmission through the novel prostacyclin receptor. The widespread expression of the prostacyclin receptor in the central nervous system suggests that prostacyclin has important roles in neuronal activity.

Animals↗
PubMed1996

Receptor imaging technique with 11C-labeled receptor ligands in living brain slices: its application to time-resolved imaging and saturation analysis of benzodiazepine receptor using [11C]Ro15-1788.

Recently we developed a novel imaging technique using positron emitter-labeled compounds as probes and a storage phosphor screen as a detector. This approach makes it possible to follow a variety of biochemical processes with spatial information in living brain slices. Further technical development is reported here in terms of time-resolved imaging and receptor characterization in a real equilibrium state. The method was validated by use of [11C]Ro15-1788, a benzodiazepine receptor antagonist. Fresh brain slices were incubated with [11C]Ro15-1788 in oxygenated Krebs-Ringer solution at 37 degrees C, in a specially designed chamber. By placing the chamber on a storage phosphor screen, we could obtain two-dimensional images of radioactivity in the slices. Time-resolved imaging was made at 5 min intervals, revealing that it took 60 min to reach equilibrium binding. The dissociation process was observed by adding an excess amount of unlabeled Ro15-1788 to the chamber, 25 min was required for the full dissociation. In the equilibrium state, i.e. in the presence of free radio-ligand, Scatchard plot analysis was performed on the cerebral cortex (Kd = 7.4 nM, Bmax = 146 fmol/mg tissue) and striatum (Kd = 7.5 nM, Bmax = 107 fmol/mg tissue), suggesting the presence of a single component of binding site in these two regions. The present method, for the first time, made it possible to study a ligand-receptor interaction in living brain slices with temporal and spatial resolutions. This technique should prove useful for studies of receptor function under physiological conditions.

Animals↗
PubMed1995

Age-related attenuation of HSP47 heat response in fibroblasts.

The collagen-binding heat shock protein of molecular weight 47,000 (HSP47), resident in the endoplasmic reticulum (ER), is assumed to play a specific role as a molecular chaperon in the processing of procollagen molecules. The present investigation of age-related alteration in the HSP47 heat response in cultured murine and human fibroblasts revealed expression in cells with a low population doubling level (PDL) derived from young mice and people more inducible by heat treatment than those from older mice and people. On the other hand, cells with a high PDL showed a very low heat response in terms of HSP47 expression regardless of the donor age. Northern blot analysis of HSP47 m-RNA indicated that the age related attenuation of HSP47 expression was regulated by transcriptional mechanisms. Furthermore, immunofluorescent analysis using a monoclonal antibody against the carboxylterminal propeptide of type I procollagen revealed far greater retention of procollagen molecules in the ER lumen of cells from old persons than in those from young persons. This was particularly prominent in heat-treated cells from old persons, indicating the possibility that the observed decrease in HSP47 heat response might cause blockage of procollagen transport to the Golgi and therefore secretion.

Aging↗
PubMed1995

In vitro positron emission tomography (PET): use of positron emission tracers in functional imaging in living brain slices.

Positron-emitting radionuclides have short half-lives and high radiation energies compared with radioisotopes generally used in biomedical research. We examined the possibility of applying positron emitter-labeled compounds to functional imaging in brain slices kept viable in an oxygenated buffer solution. Brain slices (300 microns thick) containing the striatum were incubated with positron emitter-labeled tracers for 30-45 min. The slices were then rinsed and placed on the bottom of a Plexiglas chamber filled with oxygenated Krebs-Ringer solution. The bottom of the chamber consisted of a thin polypropylene film to allow good penetration of beta+ particles from the brain slices. The chamber was placed on a storage phosphor screen, which has a higher sensitivity and a wider dynamic range than X-ray films. After an exposure period of 15-60 min, the screen was scanned by the analyzer and radioactivity images of brain slices were obtained within 20 min. We succeeded in obtaining quantitative images of (1) [18F]fluorodeoxyglucose uptake, (2) dopamine D2 receptor binding, (3) dopa-decarboxylase activity, and (4) release of [11C]dopamine preloaded as L-[11C]DOPA in the brain slice preparation. These results demonstrate that positron emitter-labeled tracers in combination with storage phosphor screens are useful for functional imaging of living brain slices as a novel neuroscience technique.

Animals↗
PubMed1994

Brain activation study by use of positron emission tomography in unanesthetized monkeys.

A system for the measurement of brain activity in conscious monkeys by positron emission tomography (PET) was established in the present study. The signal/noise ratio was maximal around 40 s for data acquisition in the PET scan with 15O-labeled water. When the monkey was stimulated by vibration and subtraction images of the data sets from regional cerebral blood flow (rCBF) changes in paired stimulation and control were superimposed on magnetic resonance images obtained from the same specimens, a somatotopic map corresponding to the sites stimulated was clearly demonstrated. Visual stimulation with a photic stimulator activated the corresponding regions of the primary visual cortex. Comparison of the activated sites and extents under the conscious state with those under anesthesia assured that the study is controllable; there was little unpredictable activation due to unlimited subject movement or to psychological effects.

Animals↗
PubMed1994

Differential induction of mRNA species encoding several classes of stress proteins following focal cerebral ischemia in rats.

We report here the time-dependent expression of several classes of HSP mRNAs following focal cerebral ischemia in rats. HSP70, GRP78, HSP27, HSP90 and HSP47 have been reported to possess distinct functions under normal and/or stress conditions. These different classes of HSP mRNAs were differentially induced by ischemia, as determined by Northern blot analysis. Messenger RNAs of the HSP70 family proteins were induced within 4 h after ischemia and then rapidly decreased, whereas HSP27 and HSP47 mRNAs reached a maximum level of expression at 24 h and 48 h after ischemic treatment, respectively. In situ hybridization showed that the expression of inducible HSP70 mRNA was observed predominantly in regions adjacent to the ischemic core except during the early periods of ischemia. HSP27 mRNA was expressed over a broad area of the ipsilateral cerebral neocortex except for the ischemic center 24 h after ischemia. The unique induction kinetics for each HSP mRNA species may reflect their distinct roles in the brain during various physiological stresses. We will also discuss that stress proteins may be involved in the central nervous system after ischemia in two important aspects: early protection against stress and restoration of damaged lesions in the brain at later stages after ischemia.

Animals↗
PubMed1994

Alternative 5' splice site selection induced by heat shock.

The mouse HSP47 gene consists of six exons separated by five introns. Three HSP47 cDNAs differing only in their 5' noncoding regions have been reported. One of these alternatively spliced mRNAs was detected only after heat shock, which caused an alternative 5' splice donor site selection. Other stress inducers, including an amino acid analog and sodium arsenite, had no effect on the alternative splicing. The alternatively spliced mRNA, which was 169 nucleotides longer in the 5' noncoding region compared to mRNA transcribed in non-heat shock conditions, was efficiently translated under heat shock conditions. This novel finding that alternative splicing is caused by artificial treatment like heat shock will provide a useful in vivo model for understanding the exon-intron recognition mechanism as well as heat shock-induced alterations in gene expression.

3T3 Cells↗