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R Kato

Publications and source records attributed to R Kato.

At least 73 records · Page 4Linked to original sources
PubMed2000

A new assay for thyroglobulin concentration in serum using monoclonal antibodies against synthetic peptides.

The concentration of thyroglobulin (Tg) measured by radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA) is greatly affected by the presence of anti-Tg autoantibodies in sera. We developed a new assay for detecting Tg in the presence of high concentrations of anti-Tg autoantibodies. A 48-kDa fragment was purified from Tg after treatment with V8 protease. This fragment did not appear to bind to two types of monoclonal antibodies (57Ab and 28D3) against a peptide in the C-terminus (amino acids 2735-2748) of Tg and intact Tg, respectively, by ELISA and Western blot analysis. In contrast, anti-Tg autoantibody or anti-Tg polyclonal antibody reacted well with this fragment. Our new ELISA used 57Ab as a solid phase antibody and 28D3 as a antibody conjugated to horseradish peroxidase. Buffer containing purified 48-kDa fragment was used to neutralize autoantibodies against Tg. With this assay, the recovery of Tg was 84.0-89.6% in normal healthy donors (n=5) in the presence of immunoglobulin G (IgG) purified from sera positive for anti-Tg autoantibody, and 76.2-104.4% in patient sera Grave's disease (n=15). Furthermore, the Tg concentrations in sera from patients with Grave's disease (n=20) ranged from 25 to 526 ng/ml, even though the Tg concentration, as measured by a commercial RIA did not exceed 55 ng/ml. There was good agreement between Tg concentrations measured by new Tg-ELISA and commercial Tg-RIA in sera that were negative for anti-Tg autoantibody. Overall, our new ELISA containing a Tg fragment to neutralize the presence of autoantibodies, showed good sensitivity and precision, and may be useful for routine use. Further investigations with the new assay should allow wider assessment of the prevalence and pattern of thyroid autoimmunity or thyroid neoplasms.

Amino Acid Sequence↗
PubMed2000

Combined therapy with hypothermia and anticytokine agents in influenza A encephalopathy.

Two children with influenza A-related encephalopathy were treated with a combination of mild hypothermia (deep body temperature of the forehead: 35 degrees C) and anticytokine agents (high-dose methylprednisolone and ulinastatin), while receiving amantadine. One of the cases exhibited acute necrotizing encephalopathy on computed tomography (CT). Although no severe complications occurred, correctable hypokalemia and hyperglycemia occurred in both cases. Both patients recovered without any neurological sequelae. Our therapeutic protocol appears to be effective for managing influenza A-related encephalopathy.

Amantadine↗
PubMed2000

The perfusion defect seen with SPECT in West syndrome is not correlated with seizure prognosis or developmental outcome.

We used interictal single photon emission computed tomography (SPECT) on 40 patients with West syndrome to determine whether cortical perfusion abnormalities are closely related to the development of West syndrome and whether they are correlated with the long-term seizure prognosis or the developmental outcome. Localized cortical perfusion abnormalities were seen in 24 patients (60%), while 15 patients (38%) were classified as normal. The remaining patient showed hyperperfusion of the basal ganglia bilaterally. Of 24 patients with localized perfusion abnormalities, unifocal cortical hypoperfusion was present in 11, multifocal hypoperfusion in 10, multiple cortical hypo- and hyperperfusion in one, hyperperfusion of the bilateral frontal cortices and brain stem in one, and focal hyperperfusion in the residual frontal cortex in one. For statistical analysis, we focused on 26 patients (cryptogenic; 10, symptomatic; 16), who were followed for more than 2 years after the onset of tonic spasms (mean 5.0 years). The results showed that focal cortical perfusion abnormalities were not correlated with the long-term seizure prognosis, the developmental outcome, or the response to ACTH therapy. In agreement with previous reports, the results of interictal SPECT suggested that focal cortical lesions play an important role in the development of West syndrome. However, statistical analysis showed that the existence of cortical dysfunction as defined by SPECT did not predict the seizure prognosis or the developmental outcome.

Cerebral Cortex↗
PubMed2000

Induction of protein kinase Czeta-related protein kinase by growth suppression in carcinogen-initiated epidermal cell-line WYF31 cells.

In primary cultured mouse epidermal cells, protein kinase C isozyme zeta (PKCzeta) consists of multiple forms, for example, low-salt eluted PKCzeta (1-PKCzeta; 79 and 85 kDa) and high-salt eluted PKCzeta (h-PKCzeta; 79 and 85 kDa) on anion-exchange column chromatography. In this study, biochemical and biophysical differences between 1-PKCzeta and h-PKCzeta were examined by using carcinogen-initiated mouse epidermal cell-line WYF31 cells, whose growth is stimulated by tumour promoter phorbol 12-myristate 13-acetate (PMA). The binding efficiency of h-PKCzeta to anti-PKCzeta antibody-affinity column was 10 times higher than that of 1-PKCzeta. T7-tagged rat PKCzeta overexpressed in WYF31 cells was recovered only in the high-salt eluted area on the anion-exchange column. Furthermore, when rat PKCzeta was stably overexpressed in WYF31 cells, the content of h-PKCzeta increased 4 to 5 times compared to that of parental cells, but the content of 1-PKCzeta was not altered. All of these results indicate that h-PKCzeta is the product of the PKCzeta gene (referred to as PKCzeta) and that 1-PKCzeta is closely related but different from PKCzeta (referred to as PKCzeta-related kinase). Interestingly, serum starvation of WYF31 cells caused a marked increase of the content of PKCzeta-related kinase with a concomitant decrease of PKCzeta content. These changes were reversed by stimulating the cell growth with 10% foetal calf serum. Prolonged treatment of starved cells with PMA, which induces the proliferation of WYF31 cells, also caused the downregulation of PKCzeta-related kinase. These results suggest that the expression levels of PKCzeta-related kinase and PKCzeta are differently regulated, and that the increased expression of PKCzeta-related kinase might play a significant role in the growth-suppression processes of WYF31 cells.

Amino Acid Sequence↗
PubMed2000

Parahisian radiofrequency catheter ablation in dogs: comparison of the above-valve and below-valve approaches.

In patients with an accessory pathway close to the His bundle, radiofrequency catheter ablation (RFCA) requires additional care to avoid damage to the normal conduction system. To assess differences between approaches from above or below the tricuspid valve (TV), we performed RFCA in 20 dogs (from above, group A, n=10; from below, group B, n=10). RF energy with temperature control at 60 degrees 60 seconds was administered at the site where a small His potential was recorded from the ablation catheter guided by fluoroscopy and transesophageal echocardiography (TEE) (in the latter six dogs). Before and after RFCA, electrophysiological testing was performed and histological findings were compared. An ablated lesion was created in 7 of 10 (2 of 2 guided by TEE) dogs in group A and 5 of 10 (3 of 4 TEE) dogs in group B. In group A, an ablated lesion involved the atrium and ventricle in the anterior site of His bundle, but the lesion was only in the ventricle in group B. An atrioventricular block (AVB) and severe damage to the penetrating bundle was observed in one dog of group A. A large hematoma on the TV was made in 2 dogs and the complete right bundle branch block (CRBBB) occurred in 3 dogs of group B. The approach from below the TV was safer than that from above the TV in parahisian RFCA, because it did not create an AVB, although it has a high incidence of CRBBB and associated technical difficulties.

Animals↗
PubMed2000

The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75 degrees C.

The radA gene is an archaeal homolog of bacterial recA and eukaryotic RAD51 genes, which are critical components in homologous recombination and recombinational DNA repair. We cloned the radA gene from a hyperthermophilic archaeon, Pyrobaculum islandicum, overproduced the radA gene product in Escherichia coli and purified it to homogeneity. The purified P. islandicum RadA protein maintained its secondary structure and activities in vitro at high temperatures, up to 87 degrees C. It also showed high stability of 18.3 kcal.mol-1 (76.5 kJ.mol-1) at 25 degrees C and neutral pH. P. islandicum RadA exhibited activities typical of the family of RecA-like proteins, such as the ability to bind ssDNA, to hydrolyze ATP in a DNA-dependent manner and to catalyze DNA strand exchange. At 75 degrees C, all DNAs tested stimulated ATPase activity of the RadA. The protein exhibited a break in the Arrhenius plot of ATP hydrolysis at 75 degrees C. The cooperativity of ATP hydrolysis and ssDNA-binding ability of the protein above 75 degrees C were higher than at lower temperatures, and the activation energy of ATP hydrolysis was lower above this break point temperature. These results suggest that the ssDNA-dependent ATPase activity of P. islandicum RadA displays a temperature-dependent capacity to exist in two different catalytic modes, with 75 degrees C being the critical threshold temperature.

Adenosine Triphosphatases↗
PubMed2000

Impairment of mycophenolate mofetil absorption by iron ion.

OBJECTIVE: We sought to evaluate the effect of iron ion on the absorption of mycophenolate mofetil, which is an immunosuppressive agent. The pharmacokinetics of mycophenolic acid were studied. METHODS: A randomized crossover design with two phases was used. A 7-day washout period separated the two treatment conditions. In the first phase, the volunteers received 1.0 g of mycophenolate mofetil alone (study 1); in the second phase, the volunteers received 1.0 g of mycophenolate mofetil and 2 tablets of iron ion preparations concomitantly (study 2). The serum concentration of mycophenolic acid, which is a pharmacologically active metabolite, was measured by reverse-phase HPLC. RESULTS: The area under the plasma concentration-time curve from 0 to 12 hours and the maximum concentration of mycophenolic acid in study 2 were significantly less than in study 1 (area under the curve, 32.9 +/- 14.7 versus 2.92 +/- 0.883 microg x h/mL, P < .001, maximum concentration, 20.1 +/- 9.21 versus 1.30 +/- 0.367 microg x h/mL, P < .001). CONCLUSIONS: This finding shows that when mycophenolate mofetil and iron ion preparations were administered concomitantly, a remarkable decrease of mycophenolate mofetil absorption was observed. Therefore it seems to be clear that we must avoid the concomitant administration of mycophenolate mofetil and iron ion preparations.

Adult↗
PubMed2000

Fentanyl reduces infarction but not stunning via delta-opioid receptors and protein kinase C in rats.

Langendorff rat hearts were used (i) to examine whether fentanyl reduces stunning, infarction or both, and (ii) to investigate if this protection is mediated by delta-opioid receptors and/or protein kinase C (PKC). In the stunning study, hearts were subjected to global ischaemia (20 min) and reperfusion. This did not produce infarction. Postischaemic mechanical function was measured in hearts treated with or without fentanyl (740 nM). Fentanyl did not affect postischaemic mechanical function. In the infarction study, the left anterior descending coronary artery was occluded for 35 min and infarct size was assessed by triphenyltetrazolium chloride staining. Hearts in the control group exhibited an infarct zone/area at risk (I/R) of 39 (SEM 5)%, whereas the I/R for the fentanyl group was 13 (2)%. When the hearts were treated with a delta-opioid receptor antagonist (naltrindole 1 nM) or a PKC inhibitor (chelerythrine 2 microM), the effect of fentanyl was abolished, with I/R of 37 (1) and 36 (2)% respectively. In our model, we conclude that fentanyl protects against infarction but not against stunning, and that the limitation of ischaemic injury is mediated by both delta-opioid receptors and PKC.

Analgesics, Opioid↗
PubMed2000

Fentanyl protects the heart against ischaemic injury via opioid receptors, adenosine A1 receptors and KATP channel linked mechanisms in rats.

We have investigated if fentanyl protects against myocardial ischaemic injury and if so, if the mechanism of this protection is mediated via opioid and adenosine A1 receptors, and KATP channels. Langendorff rat hearts were subjected to global ischaemia (30 min) and reperfusion (60 min). The drugs were administered before induction of ischaemia and maintained throughout the experiment. Treatment with fentanyl 740 nmol litre-1 improved post-ischaemic mechanical function, assessed as developed pressure, +dP/dtmax and -dP/dtmin, compared with controls after 60 min of reperfusion. These effects were abolished by naloxone 1 mumol litre-1, DPCPX 10 mumol litre-1, a selective adenosine A1 antagonist and sodium 5-hydroxydecanoate 100 mumol litre-1, a K+ATP channel blocker. We conclude that fentanyl protected the heart against post-ischaemic injury by a mechanism which was blocked by an opioid and an adenosine A1 receptor antagonist and also by a KATP channel antagonist.

Adenosine Triphosphate↗
PubMed2000

Crystallization and preliminary X-ray crystallographic studies of Thermus thermophilus HB8 MutM protein involved in repairs of oxidative DNA damage.

MutM protein, which removes the oxidatively damaged DNA base product, 8-oxoguanine (GO), has been crystallized by means of a hanging-drop vapor-diffusion procedure using polyethyleneglycol monomethylether 2000 as a precipitant in 2-(cyclohexylamino) ethanesulfonic acid (CHES) buffer, pH 9.8. The diffraction data derived from oscillation photographs indicate that the crystals belong to the monoclinic system and space group P2(1). The crystals have unit-cell dimensions of a = 45.4 A, b = 62.0 A, c = 99.7 A, and beta = 90.8 degrees. Assuming that the asymmetric unit contains two molecules, the Vm value was calculated to be 2.35 A(3).Da(-1). The crystals diffracted X-rays to at least 2.1 A resolution and were suitable for high-resolution X-ray crystal structure determination.

Crystallization↗
PubMed2000

Mechanisms of Igf2/H19 imprinting: DNA methylation, chromatin and long-distance gene regulation.

The mouse Igf2 and H19 genes lie 70-kb apart on chromosome 7 and are reciprocally imprinted. Two regulatory regions are important for their parental allele-specific expression: a differentially methylated region (DMR) upstream of H19 and a set of tissue-specific enhancers downstream of H19. The enhancers specifically activate Igf2 on the paternal chromosome and H19 on the maternal chromosome. The interactions between the enhancers and the genes are regulated by the DMR, which works as a selector by exerting dual functions: a methylated DMR on the paternal chromosome inactivates adjacent H19 and an unmethylated DMR on the maternal chromosome insulates Igf2 from the enhancers. These processes appear to involve methyl-CpG-binding proteins, histone deacetylases and the formation of chromatin insulator complexes. The Igf2/H19 region provides a unique model in which to study the roles of DNA methylation and chromatin structure in the regulation of chromosome domains.

Animals↗
PubMed2000

Comparative genomic sequencing identifies novel tissue-specific enhancers and sequence elements for methylation-sensitive factors implicated in Igf2/H19 imprinting.

A differentially methylated region (DMR) and endoderm-specific enhancers, located upstream and downstream of the mouse H19 gene, respectively, are known to be essential for the reciprocal imprinting of Igf2 and H19. To explain the same imprinting patterns in non-endodermal tissues, additional enhancers have been hypothesized. We determined and compared the sequences of human and mouse H19 over 40 kb and identified 10 evolutionarily conserved downstream segments, 2 of which were coincident with the known enhancers. Reporter assays in transgenic mice showed that 5 of the other 8 segments functioned as enhancers in specific mesodermal and/or ectodermal tissues. We also identified a conserved 39-bp element that appeared repeatedly within the DMR and formed complexes with specific nuclear factors. Binding of one of the factors was inhibited when the target sequence contained methylated CpGs. These complexes may contribute to the presumed boundary function of the unmethylated DMR, which is proposed to insulate maternal Igf2 from the enhancers. Our results demonstrate that comparative genomic sequencing is highly efficient in identifying regulatory elements.

5' Untranslated Regions↗
PubMed2000

[Influenza virus surveillance in Tama, Tokyo--viral examinations in 1996/97, 1997/98, 1998/99, and 1999/2000 seasons].

During the 1996/97, 1997/98, 1998/99, and 1999/2000 seasons, isolations of influenza virus and/or confirmations of hemagglutinin gene of virus from the throat swab of patients in 9 medical institutions in Tama, Tokyo were carried out by RT-PCR and tissue culture with MDCK cells. In 1996/97 and 1997/98 seasons, type A (H3N2) was the predominant type of influenza virus, which was 85% in the 1996/97 and 97% in the 1997/98, but small-scale outbreaks with type B virus was also confirmed in the 1996/97 season (during the 8th and 10th week). On the other hand, in the other two seasons, 2 different kinds of types of virus were discovered, which was type A (H3N2) and type B in 1998/99 season, and H3N2 and H1N1 of type A virus in 1999/2000 season. In the 1998/99 season, type A (H3N2) prevailed in the first half, and type B gradually replaced type A (H3N2) after the 5th week in 1999, whereas 2 different kinds of type A in the 1999/2000 mixed during the outbreak. From the presented data, the difference of mode by age groups and year could be explained. Namely, most specimens and positive cases in 1996/97 and 1997/98 seasons were those from low age groups (fifteen years of age and below). In 1998/99 and 1999/2000 seasons, the number of specimens from high age groups (sixteen years of age or over) accounted for above 50%, and about 70% of type A (H3N2)-positive cases. Contrary, the frequency of type B virus in 1998/99 season and type A (H1N1) in 1999/2000 season was higher in the low age groups than the high ones.

Hemagglutination, Viral↗
PubMed2000

Structure-activity relationship of orally potent tripeptide-based HIV protease inhibitors containing hydroxymethylcarbonyl isostere.

We designed and synthesized a new class of peptidomimetic human immunodeficiency virus protease inhibitors containing a unique unnatural amino acid, allophenylnorstatine [Apns; (2S,3S)-3-amino-2-hydroxy-4-phenylbutyric acid], with a hydroxymethylcarbonyl isostere as the active moiety. From a structure-activity relationship study of HIV-1 protease inhibition, enzyme selectivity for other aspartyl proteases, the antiviral activity and pharmacokinetics in rats, 24c (KNI-227) and 24d (KNI-272, our first clinical candidate) were found to be selective and orally potent HIV protease inhibitors. Moreover, an improvement of the pharmacokinetic features of KNI-272 provided two long-lasting and highly bioavailable compounds (24g: JE-2178, 24h: JE-2179).

Animals↗
PubMed2000

Prevalence of migraine and Raynaud's phenomenon in Japanese patients with vasospastic angina.

The prevalence of migraine and Raynaud's phenomenon in Japanese patients with vasospastic angina (group I) were compared with those in 2 control groups: one with effort angina (group II) and the other group without known ischemic heart disease (group III). There were no significant differences among the 3 groups with respect to age and sex. The prevalence of migraine in group I was 23 of 100, as compared with 4 of 100 in group II (p<0.01) and 11 of 100 in group III (p<0.05). The prevalence of Raynaud's phenomenon in group I was 9 of 100, as compared with 3 of 100 in group II and 4 of 100 in group III. Thus, in Japan, the prevalence of migraine in patients with vasospastic angina was higher than those in the 2 control groups, whereas the prevalence of Raynaud's phenomenon did not differ significantly among the 3 groups. The prevalence of Raynaud's phenomenon in Japanese patients with vasospastic angina was different from that reported from North America, although the prevalence of migraine was the same. This may be partially explained by racial differences.

Aged↗
PubMed2000

Accumulation of identical T cell clones in the right and left lobes of the thyroid gland in patients with Graves' disease: analysis of T cell clonotype in vivo.

To elucidate the involvement of intrathyroidal T cells in the thyroid antigen-specific immune response in Graves' disease (GD), we investigated whether identical T cell clonotypes accumulate clonally in the right and left lobes of thyroid glands of GD patients. mRNAs extracted from thyroid glands of five females patients with GD were reverse-transcribed to cDNA and then the genes coding the T cell receptor B chain variable (V-NDN-J) region were amplified using polymerase chain reaction. Single strand conformation polymorphism analysis and subsequently nucleotide sequencing were also performed to determine the clonotype of accumulating T cells. T cells infiltrating the thyroid glands showed oligoclonal expansion. The expanded T cell clonotypes were not detected in peripheral blood of the same patients. Importantly, the majority of expanding T cell clonotypes in the two lobes of the thyroid glands were identical. Our findings suggest that the clonal expansion of identical T cell clonotypes in the two lobes is driven by factors common to both lobes, such as thyroid-specific antigens, in patients with Graves' disease.

Adult↗
PubMed2000

[Studies of gastric acid secretion and gastric mucosal change after Helicobacter pylori eradication--the short and long-term observation].

We studied the gastric acid secretion and the histological findings of gastric mucosa in 48 patients with peptic ulcer before and after successful Helicobacter pylori (H. pylori) eradication. The pH of fasting gastric juice significantly decreased from 2.46 to 1.65 four weeks after eradication (p < 0.001). The factors such as disease, atrophic border, inflammatory cell involvement and histological atrophy were analyzed by logistic regression. Only the atrophic border was a significant and independent factor to convert the acidity after eradication, and the pH of gastric juice prominently decreased in the patients with widespread atrophic mucosal area. In the 24-hour intragastric pH monitoring, the pH3 holding time overnight remarkably decreased from 54.1% to 22.3% after eradication (p < 0.005). In the long follow-up of 17 patients (the mean follow-up period was 15.2 months), the pH of gastric juice kept a lower titer of 1.66. The mean atrophic scores in the upper body reduced from 1.5 to 0.9 (p < 0.05). In addition to the functional recovery of parietal cells after eradication, it was considered that the histological improvement of atrophy in oxyntic mucosa was associated with the recovery of pH.

Adult↗