Visual illusory and hallucinatory phenomena in a patient with left occipital seizures.
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Anticonvulsants↗
Biomedical subjects
Makoto Kawai
Publications and source records attributed to Makoto Kawai.
Intracellular mechanism of the negative inotropic effect induced by alpha1-adrenoceptor stimulation in mouse myocardium.
Alpha(1)-adrenoceptor stimulation (alpha(1)ARS) shows a positive inotropic effect in most mammalian myocardium. In mouse myocardium, however, alpha(1)ARS showed the negative inotropic effect, of which intracellular mechanisms are not fully clarified. The purpose of this study is to investigate the intracellular mechanism of the negative inotropic effect by alpha(1)ARS in C57BL/6 mouse myocardium. We used isolated ventricular papillary muscles of C57BL/6 strain mouse which is widely used for genetic manipulation. We simultaneously measured isometric tension and intracellular Ca(2+) concentration ([Ca(2+)](i)) using the aequorin method. In twitch contraction, phenylephrine concentration-dependently (1-100 microM) decreased tension without significant changes in the Ca(2+) transient, and these effects were completely blocked by prazosin (3 microM) or calphostin C (a PKC inhibitor, 1 microM). Phorbol 12-myristate 13-acetate (PMA) (a PKC activator, 1 microM) decreased tension as observed in phenylephrine. After PMA application, the negative inotropic effect of phenylephrine disappeared. To estimate the Ca(2+) sensitivity, tetanic contraction was produced, and the relation between [Ca(2+)](i) and tension at a steady state was measured. Phenylephrine (10 microM) decreased the Ca(2+) sensitivity, and PMA showed a similar Ca(2+) desensitizing effect. These results suggest that the negative inotropic effect of phenylephrine in mouse myocardium can be explained by the decrease in the Ca(2+) sensitivity through the activation of PKC. The present result indicates that the effect of alpha(1)ARS differs among species and strains of experiment animals. Thus, we should be careful about using the results of mouse myocardium to understand the functions of the human heart.
Animals↗
Use of the Ca-shortening curve to estimate the myofilament responsiveness to Ca2+ in tetanized rat ventricular myocytes.
We previously estimated the myofilament responsiveness to Ca(2+) in isolated intact ventricular myocytes, using the steady-state relationship between cytosolic Ca(2+) concentration ([Ca(2+)](i)) and cell-shortening during tetanus (Ca-L trajectory). This method was useful and easy; however, it could not be used for a high dose of Ca sensitizer because the instantaneous plots after the application of Ca sensitizer did not make a fixed point of shortening (we used 5% shortening). Therefore we must produce another method to investigate Ca(2+) responsiveness. For an estimation of a wider range of the Ca-L trajectory, we fitted the Ca-L trajectory data with the Hill equation to construct the Ca-shortening curve. To fit this curve, we measured the maximal shortening, which was on average 31.6%. The value of [Ca(2+)](i) to produce the half-maximal shortening (Ca(50)) was dose-dependently decreased by EMD57033 (sensitization). Either isoproterenol or 3-isobutyl-1-methylxanthine increased Ca(50) (desensitization) with a concomitant increase in intracellular c-AMP. EMD57439, a selective PDE-III inhibitor, did not significantly increase the c-AMP concentration and produced little change in Ca(50). These results are in agreement with previous reports with skinned or intact multicellular preparations. The Ca-shortening curve constructed in intact cardiac myocytes can be used to estimate the myofibrillar responsiveness to Ca(2+) in a wide range of [Ca(2+)](i).
1-Methyl-3-isobutylxanthine↗
Differential effects of left and right hemispheric seizure onset on heart rate.
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Adult↗
Incidence of type-2 diabetes mellitus in a large population of Japanese male white-collar workers.
Since occupation, a major socio-economic factor, may be a risk factor for type-2 diabetes mellitus (DM), we compared DM incidence among four groups of white-collar workers in a follow-up study in 13,547 Japanese men. The subjects aged 30-59 years and free from DM at baseline consisted of: (a) 3725 clerical, (b) 5575 technical/professional, (c) 3474 manager/administrative and (d) 774 sales workers. Incident DM was identified by 'fasting serum glucose >or=7.00 mmol/l' or 'under medical treatment for DM'. Hazard ratio (HR) and 95% confidence interval (CI) for incident DM were estimated using clerical workers as the reference group (HR=1.00). Baseline age, body mass index (BMI), drinking, smoking, exercise and education were computed as confounders. During mean follow-up periods of 7.4 years, 176 clerical, 264 technical/professional, 195 manager/administrative and 49 sales workers developed DM (6.0, 6.1, 8.8 and 9.4 per 1000 person-years, respectively). Sales workers aged 40-49 and 50-59 years had increased multivariate-adjusted HRs (95% CIs) of 1.55 (1.02, 2.35) and 2.01 (0.98, 4.10), respectively. Manager/administrative workers aged 50-59 years had an increased crude HR (95% CI) of 1.64 (1.02, 2.63), but the significance disappeared after BMI-adjustment (HR (95% CI: 1.46 (0.91, 2.35)). Technical/professional workers had no significant HR in any model. In conclusion, sales workers in Japan aged >or=40 years may have increased risk for DM (+55 to +100%) independent of BMI, lifestyles and education, and manager/administrative workers aged >or=50 years may have increased risk (about +65%) due to their large BMI.
Adult↗
InCeP: intracellular pathway based on mKIAA protein-protein interactions.
Since December 2001, we have been conducting a project to isolate and determine entire sequences of mouse KIAA cDNA clones, which encode polypeptides corresponding to human KIAA proteins. The ultimate goal of this project has been elucidation of the functions of KIAA proteins. To address this issue, we have been generating 'libraries' of antibodies against mKIAA proteins. We have, to date, already generated >800 antibodies. Using our 'libraries' of antibodies, we are now identifying endogenous mKIAA protein-protein interactions. In the present study, novel interactions were identified by MS/MS analysis following immunoprecipitation with anti-mKIAA antibodies. The interactions with biologically known molecules should enable us to predict the function of mKIAA/KIAA proteins, including hypothetical proteins identified in our cDNA project. These interactions are subsequently used for construction of an intracellular pathway related to the mKIAA protein, and the pathway is distributed through the InCeP (IntraCellular Pathway based on mKIAA protein-protein interactions) database. Users can freely access the InCeP through the internet and download the graphical display as well as the curated information.
Amino Acid Sequence↗
Policemen and firefighters have increased risk for type-2 diabetes mellitus probably due to their large body mass index: a follow-up study in Japanese men.
BACKGROUND: Policemen/firefighters may have increased risk for mortality from ischemic heart disease and from all causes. We compare incidence of type-2 diabetes mellitus (DM), a well-known predictor for mortality, in policemen/firefighters with that in two other worker-groups. METHODS: A follow-up study of 5,130 healthy Japanese men aged 30-49 years at baseline consisted of three worker-groups; 3,111 clerical workers, 1,122 manual/production and transport/communication workers, and 897 policemen/firefighters. Incident DM was identified by "fasting serum glucose >or=7.00 mmol/L" or/and "under medical treatment for DM." Hazard ratio (HR) and 95% confidence interval (95% CI) for incident DM was estimated by Cox's proportional hazard models using clerical workers as a reference group (HR=1.00). Baseline age, body mass index (BMI, kg/m(2)), drinking, smoking, exercise, and education were computed as confounders. RESULTS: During mean follow-up periods of 8.4 years, 155 clerical workers, 51 manual/production and transport/communication workers, and 74 policemen/firefighters had incident DM (5.9, 5.7, and 9.6 per 1,000 person-years, respectively; P=0.001 by chi(2)-test). Policemen/firefighters had significantly increased risk for incident DM against clerical workers (age-adjusted HR (95% CI): 1.65 (1.25, 2.18)), but the significance disappeared after adjustments for BMI (age- and BMI-adjusted HR (95% CI): 1.16 (0.87, 1.54)). Manual/production and transport/communication workers had no increased risk. CONCLUSIONS: Policemen/firefighters have increased risk for DM probably due to their large BMI. Body weight control may be more efficient for prevention of DM and DM-related health problems in policemen/firefighters than in other workers.
Adult↗
Effects of spironolactone during an angiotensin II receptor blocker treatment on the left ventricular mass reduction in hypertensive patients with concentric left ventricular hypertrophy.
BACKGROUND: Angiotensin II receptor blockers (ARB) are now commonly used to treat hypertension because of their beneficial effects on cardiovascular remodeling. However, ARB treatment can not inhibit the left ventricular (LV) remodeling sufficiently, which may be related with aldosterone secretion. To inhibit the action of aldosterone during ARB treatment, the additional effects of an aldosterone blocker and spironolactone (SPRL) on LV hypertrophy in patients with essential hypertension was studied. METHODS AND RESULTS: The patients with essential hypertension were randomly divided into 2 groups; 1 group was treated with an ARB, candesartan (8 mg/day), for 1 year (ARB group) and other group was treated with the ARB for the first 6 months and with the ARB plus SPRL (25 mg/day) for the next 6 months (combination group). Seventy patients who underwent echocardiography every 6 months were analyzed and were also classified into 4 subgroups of LV geometric pattern according to the LV mass index (LVMI) and the relative wall thickness (RWT). The ARB treatment and the addition of SPRL significantly reduced the blood pressure, however, both treatments did not affect the LV geometry in both groups. The ARB treatment in the subgroups of concentric LV remodeling (RWT>or=0.45 and LVMI<125) and concentric LV hypertrophy (RWT>or=0.45 and LVMI>or=125) significantly reduced RWT. However, ARB treatment in all subgroups did not affect LVMI. The addition of SPRL only in the concentric LV hypertrophy subgroup significantly reduced the LVMI, despite similar changes in blood pressure. CONCLUSIONS: These results indicated that the addition of SPRL treatment during the ARB treatment and conventional treatments is clinically useful to reduce the LVMI in hypertensive patients with concentric LV hypertrophy; however, does not improve the eccentric LV hypertrophy.
Aged↗
[A case of steroid-resistant acute hypersensitivity pneumonitis successfully treated with an immunosuppressive drug].
A 28-year-old man complaining of cough and fever was hospitalized because of bilateral diffuse granular lung shadows. Hypersensitivity pneumonitis was diagnosed based on bronchoalveolar lavage fluid (BALF) and transbronchial lung biopsy (TBLB). Since antigen avoidance alone was not effective, steroid pulse therapy was started, and his symptoms and chest X-ray findings improved. After discharge, he moved to another residence. A few weeks later, fever and dyspnea recurred, then he was hospitalized on the suspicion of acute exacerbation of hypersensitivity pneumonitis. Steroid therapy resulted in no improvement on this occasion. Lung biopsy under video-assisted thoracoscopy was performed, and acute hypersensitivity pneumonitis was diagnosed pathologically. Although steroid therapy was continued, hypoxia still remained and a KL-6 level markedly increased. Combined therapy with steroid and cyclosporin A was started, and his symptoms, physical findings, laboratory data, and chest X-ray findings gradually improved. There has been no report in which cyclosporin A was used for acute hypersensitivity pneumonitis but this case indicates that cyclosporin A is efficacious for its treatment.
Adult↗
[Case of carbon monoxide poisoning with the lesions demonstrated as abnormal intensity in the basal part of the left temporal lobe by MR image in acute phase].
A 46-year-old male patient was found unconscious in his room. There were remnants of burned-out charcoal briquettes. On arrival to our ER, the concentration of carbon monoxide (CO) hemoglobin was detected in 16.2%, and he was diagnosed with CO poisoning based on both his circumstantial evidence and laboratory results. Brain CT obtained on Day 1 revealed low density area in the basal part of the left temporal lobe, and T2-weighted MR images of the brain on Day 8 showed hyperintensity in the same region. MR images on Day 15 showed findings caused by white matter encephalopathy, and signal intensity in the left temporal lobe was decreased. To our best knowledge, it is very rare to see abnormal signal changes in the temporal lobe on radiological imaging of the patients who is suspected CO poisoning in acute phase because it has been reported that white matter encephalopathy after CO poisoning is commonly seen in delayed timing.
Acute-Phase Reaction↗
Presenilin 2 regulates the systolic function of heart by modulating Ca2+ signaling.
Genetic studies of families with familial Alzheimer's disease have implicated presenilin 2 (PS2) in the pathogenesis of this disease. PS2 is ubiquitously expressed in various tissues including hearts. In this study, we examined cardiac phenotypes of PS2 knockout (PS2KO) mice to elucidate a role of PS2 in hearts. PS2KO mice developed normally with no evidence of cardiac hypertrophy and fibrosis. Invasive hemodynamic analysis revealed that cardiac contractility in PS2KO mice increased compared with that in their littermate controls. A study of isolated papillary muscle showed that peak amplitudes of Ca2+ transients and peak tension were significantly higher in PS2KO mice than those in their littermate controls. PS2KO mouse hearts exhibited no change in expression of calcium regulatory proteins. Since it has been demonstrated that PS2 in brain interacts with sorcin, which serves as a modulator of cardiac ryanodine receptor (RyR2), we tested whether PS2 also interacts with RyR2. Immmunoprecipitation analysis showed that PS2, sorcin, and RyR2 interact with each other in HEK-293 cells overexpressing these proteins or in mouse hearts. Immunohistochemistry of heart muscle indicated that PS2 colocalizes with RyR2 and sorcin at the Z-lines. Elevated Ca2+ attenuated the association of RyR2 with PS2, whereas the association of sorcin with PS2 was enhanced. The enhanced Ca2+ transients and contractility in PS2KO mice were observed at low extracellular [Ca2+] but not at high levels of [Ca2+]. Taken together, our results suggest that PS2 plays an important role in cardiac excitation-contraction coupling by interacting with RyR2.
Animals↗
Temporal change in mKIAA gene expression during the early stage of retinoic acid-induced neurite outgrowth.
mKIAA genes are mouse counterparts of human KIAA genes, which were isolated in our cDNA project and were functionally unknown at the time they were sequenced. Because KIAA/mKIAA genes were isolated mainly from cDNA libraries derived from brain tissues, they are thought to be important for the organization and function of the brain. To investigate the participation of mKIAA genes in neuronal phenomena, we analyzed retinoic acid-induced neurite outgrowth using an mKIAA oligonucleotide microarray. Focusing on the early stage of this outgrowth phenomenon, we analyzed temporal gene expression changes 1-24 h after treatment with retinoic acid and found several change patterns in 38 mKIAA genes. Among them, six were upregulated at 3 h and subsequently returned to the steady state. Supposing that these genes had important roles, we performed semi-quantitative RT-PCR analysis and confirmed the existence of temporal expression patterns in two genes (mKIAA0182 and mKIAA1039). Further computational analysis of the 38 genes enabled us to find the cellular pathway associated with 6 of them with high confidence. These results indicate that some mKIAA genes are apparently relevant to retinoic acid-induced neurite outgrowth.
Animals↗
Triton-polyacrylamide gel electrophoresis and leucine aminopeptidase activity staining detect Triton-slowed bands including high-molecular-mass aminopeptidase N (CD13) isoform in cholestatic patient sera.
BACKGROUND: Western blotting of aminopeptidase N (APN) detects a high-molecular-mass isoform (260 kDa) [M. Kawai, Y. Otake, Y. Hara High-molecular-mass isoform of aminopeptidase N/CD13 in serum from cholestatic patients. Clin Chim Acta 330 (2003) 141-149] in cholestatic patient serum but is time-consuming. METHODS: Human sera were electrophoresed on polyacrylamide gel containing Triton-X100 (Triton-PAGE) and stained with leucine-B-naphthylamide (LAP-staining). The stained bands were eluted from the gel, treated with N- and O-glycosidase if necessary, and analyzed by Western blotting [M. Kawai, Y. Otake, Y. Hara High-molecular-mass isoform of aminopeptidase N/CD13 in serum from cholestatic patients. Clin Chim Acta 330 (2003) 141-149]. RESULTS: Triton-PAGE and LAP-staining clearly detected fast bands in all the sera examined. Almost parallel with leucine aminopeptidase activity, slow bands were strongly stained in all 11 cholestatic patients but clearly stained in 3 out of 14 patients with hepatobiliary diseases other than cholestasis. PAGE with various concentrations of Triton showed that Triton slows down slow bands but not fast bands. Western blotting showed that Triton-PAGE-slow bands of cholestasis contained 140 and 260-kDa APN and that fast bands were slightly smaller than monomer-size slow bands after glycosidase treatment. CONCLUSIONS: Less time-consuming than Western blotting, Triton-PAGE and LAP-staining detect novel APN bands slowed by Triton and partly composed of the high-molecular-mass isoform in cholestasis. The slow bands seem to be homodimers of APN with transmembrane anchors. The polypeptide of the fast band seems to be processed differently from that of the slow band.
Blotting, Western↗
5-Lipoxygenase inhibitors: convenient synthesis of 4-[3-(4-heterocyclylphenylthio)phenyl]-3,4,5,6-tetrahydro-2H-pyran-4-carboxamide analogues.
A convenient synthetic route to 4-[3-(4-heterocyclylphenylthio)phenyl]-3,4,5,6-tetrahydro-2H-pyran-4-carboxamide analogues as 5-LO inhibitors is described. This methodology enabled rapid development of structure-activity relationships (SARs) leading to improvement of pharmacological properties. Thus, new compounds with higher 5-LO inhibitory potency were discovered. The stereo-chemistry requirements of the tetrahydropyran ring are also discussed.
Lipoxygenase Inhibitors↗
A novel approach to protein expression profiling using antibody microarrays combined with surface plasmon resonance technology.
We have previously described our systems for the high-throughput production of antibodies against mouse KIAA proteins and their validation (Proteomics 2004, 4, 1412-1416). Using our "libraries" of antibodies, we established a novel antibody microarray system in which surface plasmon resonance (SPR) technology is utilized for signal detection. Up to 400 real-time antibody-target bindings could be measured simultaneously within a single hour. This rapid detection was achieved by direct readout of the bindings using SPR technology. To evaluate our system, we assessed the reproducibility on crude protein samples and obtained satisfactorily reproducible results, exhibiting correlation values >0.92. Using this SPR-based antibody microarray system, we examined mKIAA protein expression in five different adult mouse tissues and identified the specific tissue expression patterns of several mKIAA proteins.
Animals↗
Influences of amino acid features of glutathione S-transferase fusion proteins on their solubility.
We have previously described our strategy for high-throughput (HT) production of recombinant antigens for anti-mKIAA antibody generation, which involves using shotgun fragments generated during entire sequencing of mKIAA cDNAs. We applied this strategy to 1628 mouse KIAA (mKIAA) cDNA fragments, and 84.2% of the GST-mKIAA fusion proteins were successfully purified. The solubility of the proteins was predicted by a small-scale bacterial culture, and a large-scale culture was then performed according to the expected results. Among them, 43.8% of the proteins were purified as a soluble form and 56.2% as an insoluble form. The average yield of the soluble proteins was 0.15 nmol/mL of bacterial culture, and that of the insoluble proteins was 0.55 nmol/mL Statistical analysis of the data revealed a significant correlation between amino acid features of the recombinant proteins and their solubility. To achieve the most effective and feasible protein expression, we constructed a decision tree in which the analyzed data were reflected. The information described here may provide practical guidelines for HT production of recombinant proteins.
Amino Acid Sequence↗
Influence of the 3'-UTR-length of mKIAA cDNAs and their sequence features to the mRNA expression level in the brain.
We have previously described the sequence features of approximately 1500 mouse KIAA (mKIAA) genes in comparison with those of human KIAA genes (Okazaki, N., Kikuno, R., Inamoto, S., Hara, Y., Nagase, T., Ohara, O., and Koga, H. 2002, DNA Res., 9, 179-188; Okazaki, N., Kikuno, R., Ohara, R., Inamoto, S., Aizawa, H., Yuasa, S., Nakajima, D., Nagase, T., Ohara, O., and Koga, H. 2003, DNA Res., 10, 35-48; Okazaki, N., Kikuno, R., Ohara, R., Inamoto, S., Koseki, H., Hiraoka, S., Saga, Y., Nagase, T., Ohara, O., and Koga, H. 2003, DNA Res., 10, 167-180; and Okazaki, N., F-Kikuno, R., Ohara, R., Inamoto, S., Koseki, H., Hiraoka, S., Saga, Y., Seino, S., Nishimura, M., Kaisho, T., Hoshino, K., Kitamura, H., Nagase, T., Ohara, O., and Koga, H. 2004, DNA Res., 11, 205-218). To validate the orthologous relationship between mKIAA and KIAA genes in detail, we examined their chromosomal positions and evolutionary rate of synonymous substitutions and confirmed that >93% of the mKIAA/KIAA gene pairs are orthologous. During the sequence analysis of mKIAA genes, we found that 3'-untranslated region (3'-UTR) lengths of mKIAA and KIAA genes are extremely long. In the meanwhile, we have also examined the tissue-specific expression of approximately 1700 mKIAA genes using cDNA microarray and verified predominantly their expression in adult brain (Koga, H., Yuasa, S., Nagase, T., Shimada, K., Nagano, M., Imai, K., Ohara, R., Nakajima, D., Murakami, M., Kawai, M., Miki, F., Magae, J., Inamoto, S., Okazaki, N., Ohara, O. 2004, DNA Res., 11, 293-304). To connect these two evidences, we statistically analysed the relationship between them by using the mKIAA genes. Consequently, a positive correlation was observed between the 3'-UTR lengths and the relative expression intensities in adult brain. Furthermore, we searched sequence elements in the 3'-UTR possibly related with their expression and found some candidates regarding the brain-specific expression.
3' Untranslated Regions↗
Microarray analysis of host gene-expression during intracellular nests formation of Trypanosoma cruzi amastigotes.
The intracellular pathogens utilize numerous cellular components of host cells to advance the infection as well as to enter the host cell. Analyzing the host cellular response enables us to get a better understanding of the pathogenesis, and subsequently indicate possible therapeutic targets. We therefore analyzed gene-expression profile of NIH3T3 fibroblast cells infected by Trypanosoma, a representative intracellular pathogen similar to Leishmania, using custom-designed cDNA microarray consisting of 1,701 mKIAA cDNAs. Focusing on intracellular nest formation of Trypanosoma cruzi amastigotes, we profiled the host gene-expression at 8 days post-infection and found several degrees of change in 16 mKIAA genes. Among these genes, 10 were up-regulated and 6 were down-regulated. Assuming that these genes had important roles in the infection's progression, we performed semi-quantitative RT-PCR analysis and con-firmed the gene expression change of 4 genes. Furthermore, 5 genes were mapped on cadherin signaling pathway using pathway analysis software. These results indicate significance of the host cellular pathway in the proliferative stage of Trypanosoma cruzi amastigotes.
Animals↗