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Biomedical subjects

K Takahama

Publications and source records attributed to K Takahama.

At least 37 records · Page 2Linked to original sources

Identification of remains by sequencing of mitochondrial DNA control region.

The maternity of two newborns who were murdered and abandoned >5 and 10 years were analyzed by amplification and direct sequencing of mitochondrial DNA (mtDNA) control regions. Sequences of two hypervariable segments from each femur bone sample and the blood of the putative mother showed four mutations in hypervariable region I and two mutations in addition to two nucleotide insertions in hypervariable region II compared with the reference sequence, and all sequences were identical. The genotype of these individuals is found to be relatively rare in the Japanese population, and it was strongly suggested that both sets of newborn remains really were children of the putative mother. Sexes of the remains were determined to be female and male by amplifying a segment of the X-Y homologous gene, amelogenin. These results demonstrate that sequencing of mtDNA is a useful tool for genetic identification of aged and decomposed materials.

Amelogenin↗

Investigation of the association between infection with Helicobacter pylori and with TT virus, a novel DNA virus, in patients with gastroduodenal ulcer or ulcer scar.

OBJECTIVE: Recently, a novel DNA virus (TT virus; TTV) has been isolated. Enteric transmission is suggested as a route of transmission of TTV, with high prevalence of this virus infection in the general population, and age and geographical distributions of TTV prevalence very similar to those of Helicobacterpylori infection. We analysed an association between TTV and H. pylori infection in patients with gastroduodenal ulcer or ulcer scar. METHODS: In 181 patients with a gastroduodenal ulcer or ulcer scar (102 with a gastric lesion, 60 with a duodenal lesion, and 19 with both sites involved), specimens were cultured for H. pylori and TTV infection was sought in serum by a polymerase chain reaction. RESULTS: H. pylori infection was demonstrated in 152 patients (84.0%) and TTV was detected in 168 patients (92.8%). Patients with TTV were significantly older than those without TTV (P = 0.0001), while no age difference was observed between patients with and without H. pylori infection. No difference was apparent in the prevalence of TTV infection between patients with and without H. pylori infection, and vice versa. CONCLUSIONS: We found no association between TTV infection and H. pylori infection in patients with peptic ulcer diseases, which is consistent with a lack of association between TTV infection and peptic ulcer. However, larger studies including surveys of the general population will be required to analyse the overall association between TTV and H. pylori.

Adult↗

Non-competitive inhibition of kainate-induced currents by diethylstilbestrol in acutely isolated mouse CA1 hippocampal neurons.

The effect of a synthetic estrogen, diethylstilbestrol (DES), on kainate-induced currents was investigated in the hippocampal CA1 pyramidal neurons acutely dissociated from the mice using the nystatin-perforated patch-clamp recording configuration under voltage-clamp conditions. DES inhibited the current evoked by 100 microM kainate in a concentration-dependent manner with a half-maximum inhibitory concentration of 8.8 microM. The action of DES was voltage-independent. Since DES produced a suppression of the maximum response of the kainate concentration-response curve, the inhibition by DES of the kainate-induced current appears to be non-competitive.

Animals↗

[Survey on postmortem examination to police surgeons and emergency physicians. Possibility of physicians' assist in mass-disaster].

We conducted a questionnaire survey of police surgeons and emergency physicians, inquiring about their experience of medicolegal investigation of death and their willingness to join a death investigation team in a major disaster. The questionnaire also asked about their knowledge about and interest in the forensic specialist system established by the Japanese Society of Legal Medicine. Police surgeons were generally willing to join an investigation team only if a disaster occurred in or close to their hometown, because they could not afford more than several days away from patient care. Although many of the emergency physicians were willing to join a death investigation team, they had difficulty in doing so without permission or orders from their employer or the authorities concerned. The survey found that the percentage of aged police surgeons was increasing among those surveyed. This fact, in combination with the current emphasis of postgraduate education on specialty training, threatens to cause a substantial lack of physicians available for medicolegal investigation of death. Therefore, it is urgently necessary to establish a system of training resident and emergency physicians in medicolegal investigation of death. In addition to providing postgraduate training in medicolegal investigation of death to prospective trainees who are emergency physicians at major hospitals in potential disaster-stricken areas, the medical school should incorporate forensic medicine in postgraduate training programs so that they can actively perform death investigation on disaster victims dying before or after arrival at their hospitals. Furthermore, the forensic community should make every effort to increase the number of autopsies in each department of forensic medicine and to expand the medical examiner system throughout Japan that is currently in practice only in the Metropolis of Tokyo and Yokohama, Nagoya, Osaka and Kobe Cities in order to incorporate forensic training in the postgraduate clinical training programs that will become compulsory in 2004.

Attitude of Health Personnel↗

A sandwich enzyme immunoassay for human muscle-specific beta-enolase and its application for the determination of skeletal muscle injury.

A sensitive sandwich enzyme immunoassay for human beta-enolase was developed and used to examine beta-enolase in blood or bloodstains as a marker for the determination of skeletal muscle injury. Human beta-enolase was purified from human skeletal muscle, and then an antibody against it was prepared. Polystyrene balls coated with rabbit anti-human beta-enolase IgG were incubated with human beta-enolase and then with anti-human beta-enolase Fab'-peroxidase conjugate. Peroxidase activity bound to the polystyrene balls was assayed by fluorometry using 3-(4-hydroxyphenyl)propionic acid as a hydrogen donor. The detection limit for human beta-enolase was 2.6 pg (30 amol) per assay. The degree of cross-reaction of the sandwich enzyme immunoassay for other organs except for heart (1/10) was about 1/150 or less. Moreover, the localization of beta-enolase in various human tissues was examined by Northern blot analysis, and this confirmed that beta-enolase was expressed only in skeletal and cardiac muscle. Antigenic activity in bloodstains containing beta-enolase was recovered well after storage for 60 days at room temperature. The ratio of beta-enolase to total protein in bloodstains made from non-traumatic blood, nasal hemorrhage and menstrual blood, was within the normal range. In contrast, the ratio of beta-enolase in bloodstains from traumatic blood was obviously elevated (10-30 fold) in comparison with non-traumatic blood. Furthermore, the ratio of beta-enolase was proved to be higher in stains adhering to weapons that had passed through skeletal muscle, indicating that detection of beta-enolase in bloodstains could be used to distinguish crime weapons. These results suggest that beta-enolase is a useful marker for identification of skeletal muscle injury as well as for detecting the origin of bleeding.

Animals↗

Y chromosome DNA polymorphisms and their haplotypes in a Japanese population.

Allele frequencies of two Y chromosome-specific short tandem repeats, locus DYS385 and DYS19, in addition to the Y Alu polymorphic (YAP) insert (DYS287) were investigated in blood samples obtained from 270 unrelated Japanese in Miyazaki Prefecture (south Japan). A total of 47 genotypes in DYS385 and 6 in DYS19 were detected, and the frequency of the YAP(+) allele was found to be 35.2% (95270). The allele distributions of each locus revealed significant differences compared with those in other populations. One hundred and twelve different haplotypes were observed in the combined 55 alleles of three loci. The gene diversity values range 0.96 for DYS385 and 0.71 for DYS19, and the combination haplotype diversity value is 0.98.

Journal Article↗

Left ventricular function at rest and during bicycle exercise in normal subjects: assessment by ECG-gated myocardial perfusion SPET with 99Tcm-tetrofosmin.

99Tcm-labelled myocardial perfusion tracers allow simultaneous assessment of myocardial perfusion and left ventricular function using ECG-gated SPET. The aim of this study was to evaluate left ventricular performance during exercise by means of ECG-gated myocardial perfusion SPET. After the administration of 99Tcm-tetrofosmin (555-740 MBq), eight healthy volunteers aged 27-49 years underwent ECG-gated myocardial perfusion SPET at rest and during supine submaximal exercise (75 and 125 W), for 3 min each. Using ECG-gated SPET data, left ventricular end-diastolic volume (LVEDV) demonstrated a biphasic response during exercise (from 106.4 +/- 17.5 to 119.9 +/- 19.9 to 108.1 +/- 19.2 ml). In contrast, left ventricular end-systolic volume decreased gradually and significantly during exercise (from 47.1 +/- 11.9 to 41.5 +/- 8.9 to 36.1 +/- 10.1 ml; P < 0.05), and left ventricular ejection fraction continued to increase at higher workloads (from 56.1 +/- 6.0 to 63.0 +/- 2.7 to 67.0 +/- 4.3; P < 0.01) despite a fall in LVEDV. There was a progressive increase in cardiac output during exercise, which reached a peak of 7.2 +/- 0.9 l.min-1. We conclude that ECG-gated myocardial perfusion SPET can assess left ventricular function during exercise and may provide useful information for the evaluation of patients with ischaemic heart disease.

Adult↗

Mucolytic and antitussive effects of erdosteine.

To investigate the influence of erdosteine, a new homocysteine-derived expectorant, on airway clearance we studied the effects of the drug on the viscosity of mucin, on the mucociliary transport rate in quails, on airway secretion in rats and on the cough reflex in guinea-pigs. The active metabolite of erdosteine, M1 (10 microM to 1 mM), significantly reduced the viscosity of porcine stomach mucin. Erdosteine by itself did not reduce viscosity. Erdosteine significantly promoted mucociliary transport in quails and increased airway secretion in rats. The effect was still apparent 24h after administration. Erdosteine significantly suppressed citric acid-induced cough reflexes in guinea-pigs but did not suppress mechanical stimuli-induced cough reflexes. Erdosteine suppressed the reduction of the recovery volume of bronchoalveolar lavage fluid and albumin leakage into the fluid in citric acid-exposed guinea-pigs. These results indicate that erdosteine removes sputum by reducing its viscosity, and by promoting mucociliary transport and sustained enhancement of airway secretion. It also suppressed the chemical stimulation-induced cough reflex and plasma leakage into the airway. These results suggest that erdosteine is an excellent expectorant with several modes of action.

Animals↗

[Composition of vascular tree using moving-table MR angiography: development and preliminary clinical experience with a semi-automated program combining stacks of MR angiographic images].

Moving-table MR angiography, combining a contrast enhanced MRA approach with table movement, has recently become available. This technique allows imaging of long, longitudinal anatomical regions of vasculature. We have developed a semi-automated program combining stacks of MRA images obtained from different image stations, and applied it to the reconstruction of vascular trees in twenty-five patients with peripheral arterial occlusive disease. Using this program, continuous vascular trees, extending from the abdominal aorta to lower leg (up to 124 cm), were semi-automatically reconstructed within two minutes from six different projection angles. Extensive vascular lesions were recognized three-dimensionally by moving image display mode.

Arterial Occlusive Diseases↗

[Unusual form of ulcerative colitis].

Segmental colitis which showed the similar findings of ulcerative colitis and right-sided colitis with back-wash ileitis has been considered as the unusual form of ulcerative colitis. Today, we have few opportunities to come across those cases, because modern diagnosis of colon disease could make differential diagnosis easily with the sense of broad spectrum of IBD. We presented three cases of segmental colitis and one case of so-called indeterminate colitis associated of carcinoma in the cecum and discussed the clinical significance for diagnosis and finding the step of studying the etiology of IBD. Especially, in the third case, thirteen years old man was showing the sign of early stage of Crohn disease with non-caseating granuloma in the biopsy specimen at first, the findings of the segmental ulcerative colitis with crypt abscess were seen after six months in the second examination. To observe the detail changes of the mucosal lesion in this kind of cases will present the new idea for studying the etiology of IBD.

Adolescent↗

Sequence polymorphism of mitochondrial DNA control region in Japanese.

Sequence polymorphisms of the mitochondrial DNA (mtDNA) control region, hypervariable regions I and II, from 100 unrelated Japanese were determined by PCR amplification and direct sequencing. Sequences of 404 nucleotides for hypervariable region I and 379 nucleotides for region II were obtained. Variable sites (85 and 45) were revealed in region I and region II, respectively, as compared to the reference sequence, and a total of 96 different genetic patterns from both regions I and II were determined. A point mutation heteroplasmy was observed at the ratio of approximately 50:50 from one individual at the sequence position 151 showing a nucleotide transition from C to T. The probability of identity was estimated as 2.3% for region I, 3.9% for region II, and 1.1% combined for both regions. These results suggest that sequence polymorphism of mtDNA control region would be very useful in forensic practice as a marker for individual identification.

Base Sequence↗

Development of a new method for simultaneously evaluating mucociliary clearance and pulmonary epithelial permeability in rabbit experiments by means of 18FDG, three-dimensional positron emission tomography and rectilinear scan.

We tried to simultaneously obtain the elimination constant of mucociliary clearance and the pulmonary epithelial permeability constant after inhalation of 2-[18F]fluoro-2-deoxy-D-glucose (18FDG) solution by carrying out whole lung positron emission tomography and a rectilinear scan in rabbit experiments. The elimination constant of pulmonary epithelial permeability was obtained from the decrease in the amount of the radioactivity with time in the region of interest (ROI) confined to the lungs, trachea and tracheal cannula in the rectilinear scan. The total elimination constant of the radioactivity in the lungs was obtained from the ROI confined to the lungs in the tomography. The mucociliary clearance rate constant in the lungs was then obtained after subtracting the elimination constant of the pulmonary epithelial permeability from the total elimination constant of the 18FDG in the lungs. The mucociliary clearance constant in the trachea was calculated from the residual radioactivity in the trachea and the mucociliary clearance constant in the lungs. The mean pulmonary epithelial permeability constant was 0.0020% min(-1) obtained from the rectilinear scan. The mean mucociliary clearance constants of the lungs and the trachea were 0.0006 and 0.025% min(-1), respectively. These results indicated that the pulmonary epithelial permeability and mucociliary clearance could be evaluated simultaneously with 18FDG by using three-dimensional positron emission tomography and a rectilinear scan.

Administration, Inhalation↗

Current opinion of muco-active drug research: strategies and problems.

In general, mucoactive drugs are classified into several groups. However, since many drugs have overlapping effects, it is difficult to classify the drugs into groups based on their major actions. It has been reported that many mucoactive drugs have antioxidant effects. It is reasonable to suggest that an anti-inflammatory property is crucial to demonstrate effectiveness in a clinical context. From this point of view, we have evaluated several mucoactive drugs over two decades. Of these, we will consider the following drugs with anti-inflammatory properties: sodium aceneuramate; glucocorticoids; traditional Chinese medicines; and new cysteine derivatives. On the basis of these findings, we believe that the efforts to seek for compatible actions between glucocorticoids and oriental medicines may provide new opportunities for development of ideal mucoactive drugs with specified actions, i.e. suppression of gene expression.

Anti-Inflammatory Agents↗

A family with Liddle's syndrome caused by a new missense mutation in the beta subunit of the epithelial sodium channel.

Liddle's syndrome is an autosomal dominant form of salt sensitive hypertension caused by mutations in the beta or gamma subunit of the epithelial sodium channel. Systematic mutagenesis studies revealed that a conserved PPPXY sequence (PY motif) of the C-terminus of the alpha, beta, or gamma subunits might be involved in the regulation of the channel activity. However, only two missense mutations in the PY motif of the beta subunit have been reported to cause Liddle's syndrome. We sequenced the C-termini of the beta and gamma subunits of the epithelial sodium channel in a Japanese family clinically diagnosed as having Liddle's syndrome and found a new missense mutation in the PY motif of the beta subunit, P615S. Expression studies with P615S mutant in Xenopus oocytes resulted in an about 3-fold increase in the amiloride-sensitive sodium current compared to the wild type (p = 0.001). These findings provide further clinical evidence for the hypothesis that a conserved PY motif may be critically important for the regulation of the epithelial sodium channel.

Adult↗

Effects of SS320A, a new cysteine derivative, on the change in the number of goblet cells induced by isoproterenol in rat tracheal epithelium.

We examined the effects of SS320A ((-)-(R)-2-amino-3-(3-hydroxypropylthio)propionic acid), a new cysteine derivative, on the change in the number of goblet cells induced by isoproterenol in rat tracheal epithelium. Four types of goblet cells were characterized in tracheal epithelium according to their size and staining affinity with Alcian blue (AB) / periodic acid Schiff (PAS). When each rat was given a single daily injection of isoproterenol (0.05 mg/kg, i.p.) for 14 days, a significant increase was observed in AB/PAS-positive cells that were recognizable as goblet cells in tracheal epithelium. When SS320A (10-100 mg/kg, p.o.) or propranolol (1 mg/kg, s.c.) was administered before each injection of isoproterenol, the increase in the number of goblet cells induced by isoproterenol was significantly inhibited. There was no difference between male and female rats with regard to this inhibitory action. On the other hand, ambroxol, bromhexine, L-cysteine ethyl ester and S-carboxymethylcysteine (100 mg/kg, p.o., respectively), which are used as expectorants, had no inhibitory effects on the isoproterenol-induced change in the number of goblet cells. Four metabolites (M1-M4) of SS320A in rats also failed to inhibit the change induced by isoproterenol. These data suggest that SS320A itself may have a beneficial effect against mucus hypersecretion in chronic respiratory diseases.

Animals↗

Inhibition of glycine-induced current by morphine in nucleus tractus solitarii neurones of guinea pigs.

We studied the effect of morphine on the current induced by glycine in acutely dissociated nucleus tractus solitarii (NTS) neurones of guinea pigs, by use of the whole-cell patch clamp technique. Morphine inhibited 30 microM glycine-induced current (Igly), without affecting the current caused by 30 microM GABA. The effect of morphine was concentration-dependent, with a maximal effect at 1 mM, and reversible. The half-maximum inhibitory concentration of morphine was 30 microM. The effect of morphine was not depressed by naloxone, an opioid antagonist. Furthermore, the effect was not substantially affected by methiothepin, a 5-HT1 antagonist, ketanserin, a 5-HT2 antagonist and MDL-72222, a 5-HT3 antagonist. Morphine at 30 microM shifted the concentration-response curve for Igly to the right without affecting the maximum value. The effect of morphine on Igly showed no use-dependence. The results indicate that morphine inhibits Igly in the NTS neurones, and further suggest that morphine at the concentration used may act on the glycine receptor-ionophore complex, but not on the Cl-channel of the complex.

Animals↗

Differential effect of codeine on coughs caused by mechanical stimulation of two different sites in the airway of guinea pigs.

We studied the difference in the effects of codeine on coughs caused by mechanical stimulation to the larynx and to the bifurcation of the trachea in lightly anaesthetized guinea pigs. Mechanical stimulation to the larynx or the bifurcation of trachea caused a stable cough response. The response was reproducible over 60 min, when stimulation was repeatedly applied at 20-min intervals. No significant difference was found between the amplitudes of the responses to mechanical stimulation of the larynx and of the tracheal bifurcation. Codeine, 10, 20 and 50 mg/kg, dose dependently depressed the coughs caused by larynx stimulation. The antitussive, however, failed to depress the cough caused by stimulation to the tracheal bifurcation, although a large dose, 50 mg/kg, significantly depressed the cough. In capsaicin-treated guinea pigs, codeine at 20 mg/kg significantly depressed the cough caused by stimulation to the tracheal bifurcation. The present results suggest that cough caused by mechanical stimulation to the larynx might be more sensitive to codeine treatment than cough caused by stimulation to the bifurcation of trachea. Furthermore, it is suggested that coughs caused by mechanical stimulation to both sites might consist of at least two components as regards their pharmacological nature.

Animals↗

A sandwich enzyme immunoassay for brain S-100 protein and its forensic application.

A sensitive sandwich enzyme immunoassay for identification of brain S-100 protein in blood or bloodstains containing brain tissue is described. A polystyrene ball coated with rabbit anti-S-100 protein IgG was incubated with human S-100 protein, and then with anti-S-100 Fab'-peroxidase conjugate. Peroxidase activity bound to the polystyrene ball was assayed by fluorometry using 3-(4-hydroxyphenyl)propionic acid as the hydrogen donor. The detection limit of human S-100 protein was 0.6 pg (30 amol) per assay tube. The cross-reaction of this sandwich enzyme immunoassay to other organs was approximately 1/100 or less. Antigenic activity of S-100 protein in bloodstains containing brain extracts was detectable after storage for 36 days at room temperature. The ratio of S-100 protein to total protein (ng/mg) in bloodstains when brain tissue was mixed with normal human blood at concentrations of 5-500 mg/ml was approximately 100-fold those of other samples (liver, heart, intestine, and skeletal muscle). These results indicated that bloodstains mixed with brain tissue were clearly distinguishable from others. Thus, in forensic practice, measurement of S-100 protein or the ratio of S-100 protein to total protein is useful to identify blood and bloodstains containing brain tissue.

Blood Stains↗