Search PubMed⌕ Search

Biomedical subjects

K Okamoto

Publications and source records attributed to K Okamoto.

At least 361 records · Page 20Linked to original sources
PubMed1998

Analysis of spinocerebellar ataxia type 2 in Gunma Prefecture in Japan: CAG trinucleotide expansion and clinical characteristics.

We analyzed 13 patients with spinocerebellar ataxia type 2 (SCA2) in seven unrelated families who live in Gunma Prefecture, Japan (population approx. 2,000,000), and documented the clinical and molecular properties correlated with the CAG repeat expansion. Twelve of the 13 patients and one presymptomatic female were genetically examined, and the CAG repeat number of the expanded and normal alleles was 40.8+/-4.8 (mean+/-S.D., n=13) and 22+/-0 (n=13), respectively. The repeat size of the expanded alleles was inversely correlated with the patients' age at onset. Paternal anticipation was observed, accompanied by an increase of the CAG repeat size. The patients presented here were clinically characterized by a relatively higher frequency of slow saccades, hyporeflexia, hypotonia, and tremor. A number of peaks in the expanded allele on polyacrylamide gel electrophoresis showed the presence of cell mosaicism in SCA2 as well. In Gunma Prefecture, SCA2, Machado-Joseph disease and spinocerebellar ataxia type 6 are almost equally present and at higher frequencies than spinocerebellar ataxia type 1 and hereditary dentatorubropallidoluysian atrophy, which are rare. Thus, the difference of frequency of autosomal dominant spinocerebellar ataxias may be present in Japan.

Adolescent↗
PubMed1998

Identification of alternative splicing forms of GLT-1 mRNA in the spinal cord of amyotrophic lateral sclerosis patients.

The glutamate transporter plays an essential role in regulating glutamate levels in the synaptic cleft. It has been postulated that the dysfunction of GLT-1, one subtype of glutamate transporter, may be etiologically related to amyotrophic lateral sclerosis (ALS). Two alternative splicing forms of GLT-1 messenger RNA (mRNA) were found in the cervical spinal cord of five ALS patients and three controls. Analysis with reverse transcription-polymerase chain reaction (RT-PCR) showed that the shorter mRNA was a result of exon 8 skipping. A truncated transcript containing an intronic sequence at the 3' end of exon 7 was also demonstrated. However, the incidence of both alternative mRNAs was not different between the five ALS patients and three controls. Interestingly, the mRNA were also found in the cerebral cortex of a control subject. These results suggest that alternative splicing forms of GLT-1 mRNAs do not play a pathogenetic role in ALS but rather a physiological one in the normal spinal cord and brain.

ATP-Binding Cassette Transporters↗
PubMed1998

T-loop deletion of CDC2 from breast cancer tissues eliminates binding to cyclin B1 and cyclin-dependent kinase inhibitor p21.

The eukaryotic cell cycle is regulated by a highly conserved family of protein kinases, the cyclin-dependent kinases (CDKs). Monomeric free CDKs do not possess enzymatic activity, largely due to the steric hindrance caused by the T-loop at the entrance of the catalytic cleft, making ATP inaccessible to the substrate. Binding of a cyclin, primarily to the NH2-terminal lobe of the CDK that surrounds the PSTAIRE helix, induces a large conformational change in the PSTAIRE helix of the CDK and also causes the T-loop to move out of the way of the catalytic cleft. We identified from breast cancer tissues a novel variant of human CDC2, termed CDC2deltaT, that lacks 171 nucleotides corresponding to 57 amino acids, which compose most of the T-loop. CDC2deltaT was detected in 10 of 14 breast cancer tissues analyzed, whereas it was not detectable in diploid human fibroblast cell lines or in interleukin 2-stimulated normal human lymphocytes. CDC2deltaT protein is unable to complex with cyclin B1 and lacks histone H1 kinase activity. CDC2deltaT also fails to bind to the CDK inhibitor p21. These results indicate that the T-loop not only plays a key role in keeping a free CDK in its inactive state but also in facilitating CDK activation by promoting cyclin binding.

Alternative Splicing↗
PubMed1998

Lithium inhibits the reverse tolerance and the c-Fos expression induced by methamphetamine in mice.

To elucidate the mechanism of psychostimulant-induced reverse tolerance, the effects of lithium on ambulatory activity and cerebral c-Fos protein expression were investigated in mice injected with methamphetamine (2 mg/kg, s.c., 1-5 times). The ambulatory activity enhanced by either acute or chronic methamphetamine injection was delayed or diminished by LiCl pretreatment (170 mg/kg, s.c., 1 h before methamphetamine). The c-Fos expression in the dorsal lateral geniculate nucleus and in the striatum was significantly increased by acute but not chronic injection of methamphetamine, and the increases were significantly suppressed by LiCl pretreatment. Although how the Li-sensitive c-Fos expressions in the dorsolateral geniculate nucleus and striatum are related to methamphetamine-induced behavioral excitation is unclear, these results suggest that lithium at least functionally interferes with the formation of the state of reverse tolerance to methamphetamine in the mouse.

Animals↗
PubMed1998

Targeted chemotherapy in mice with peritoneally disseminated gastric cancer using monoclonal antibody-drug conjugate.

The murine monoclonal antibody A7 (MAb A7) is reactive against most human gastric cancer cell lines. Using a nude mouse peritoneal dissemination model of human gastric cancer, we investigated targeted chemotherapy using a conjugate of neocarzinostatin (NCS) with MAb A7 (A7-NCS). After demonstrating cytotoxicity of the complex against the human gastric cancer cell line MKN45 in vitro, we intraperitoneally injected A7-NCS, NCS or saline into nude mice bearing peritoneally disseminated human gastric cancer. A7-NCS inhibited peritoneal dissemination significantly more effectively than NCS. MAb A7 may prove to be an effective carrier for antineoplastic drugs in patients with peritoneal dissemination of gastric cancer.

Animals↗
PubMed1998

Characterization of physical entrapment and chemical conjugation of adriamycin in polymeric micelles and their design for in vivo delivery to a solid tumor.

An anticancer drug adriamycin (ADR) was incorporated into polymeric micelles forming from poly(ethylene glycol)-poly(aspartic acid) block copolymer by chemical conjugation and physical entrapment. Structural stability of the polymeric micelles was found to be dependent on both the contents of chemically conjugated and physically entrapped ADR. The polymeric micelle with high contents of the chemically conjugated ADR and the physically entrapped ADR expressed very high in vivo antitumor activity against murine C 26 tumor, while the polymeric micelle with only the chemically conjugated ADR showed negligible in vivo activity. This indicates that the physically entrapped ADR played a major role in antitumor activity in vivo. For the polymeric micelle with the high ADR contents, it was found that a dimer of adriamycin molecules formed and that this dimer was physically entrapped in the inner core of the micelle as well as intact ADR.

Animals↗
PubMed1998

Multiple myeloma associated with serum amino acid disturbance and high output cardiac failure.

We experienced a plasma cell leukemia (PCL) patient complicated with high output cardiac failure (HOCF), proved as his elevated cardiac index and pulmonary artery wedge pressure and decreased systemic vascular resistance index in a hemodynamic study. We found no possible causes of HOCF. Interestingly, HOCF was improved as PCL responded to intensive chemotherapy. On the other hand, he showed consciousness disturbance, and had frequent attacks of generalized seizure. His electroencephalogram showed slow waves, and a spike and wave complex. Hyperammonemia and abnormal amino acid distribution were also found. This abnormal serum amino acid distribution, especially elevated glycine level, was different from that seen in chronic liver failure, and he had no hepatic disease. After intensive chemotherapy, the serum ammonia level and glycine level decreased. In this patient, PCL seemed to be responsible for HOCF, hyperammonemia, and abnormal amino acid distribution. We experienced two more cases of multiple myeloma (MM) with HOCF, hyperammonemia, abnormal serum amino acid distribution, and consciousness disturbance of unknown origin. Those two cases showed slow waves in the electroencephalogram. Improvement was seen in their HOCF, hyperammonemia, and abnormal amino acid levels after chemotherapy. The possibility of MM as a cause of HOCF is discussed.

Adult↗
PubMed1998

Evidence for autoantigens of Env/Tax proteins in human T cell leukemia virus type I Env-pX transgenic mice.

OBJECTIVE: To examine T cell clonotypes infiltrating into arthritic joints and to investigate whether human T cell leukemia virus type I (HTLV-I) env-pX gene products act as autoantigens in HTLV-I env-pX transgenic mice. METHODS: Complementary DNA (cDNA) encoding the V-D-J (third complementarity-determining region [CDR3]) region of T cell receptor beta chain was amplified by Vbeta family polymerase chain reaction. T cell clonotypes were detected by a single-strand conformational polymorphism method, and sequence analysis of the CDR3 region was performed. RESULTS: Distinct oligoclonal T cell expansion was observed in arthritic joints, and a conserved amino acid motif in the CDR3 region was found in T cells infiltrating joints. Moreover, several intraarticular T cells recognized HTLV-I Env and Tax proteins. CONCLUSION: Our results suggest that HTLV-I Env and Tax proteins act as autoantigens that are recognized by autoreactive T cells in inflamed arthritic lesions in the HTLV-I env-pX transgenic mouse. Thus, some T cells infiltrating the joint recognize Env or Tax protein. These cells may trigger chronic arthritis in HTLV-I env-pX transgenic mice.

Amino Acid Sequence↗
PubMed1998

Longitudinal study of cerebrospinal fluid levels of tau, A beta1-40, and A beta1-42(43) in Alzheimer's disease: a study in Japan.

To clarify the alterations of tau, amyloid beta protein (A beta) 1-40 and A beta1-42(43) in the cerebrospinal fluid (CSF) that accompany normal aging and the progression of Alzheimer's disease (AD), CSF samples of 93 AD patients, 32 longitudinal subjects among these 93 AD patients, 33 patients with non-AD dementia, 56 with other neurological diseases, and 54 normal control subjects from three independent institutes were analyzed by sensitive enzyme-linked immunosorbent assays. Although the tau levels increased with aging, a significant elevation of tau and a correlation between the tau levels and the clinical progression were observed in the AD patients. A significant decrease of the A beta1-42(43) levels and a significant increase of the ratio of A beta1-40 to A beta1-42(43) were observed in the AD patients. The longitudinal AD study showed continuous low A beta1-42(43) levels and an increase of the ratio of A beta1-40 to A beta1-42(43) before the onset of AD. These findings suggest that CSF tau may increase with the clinical progression of dementia and that the alteration of the CSF level of A beta1-42(43) and the ratio of A beta1-40 to A beta1-42(43) may start at early stages in AD. The assays of CSF tau, A beta1-40, and A beta1-42(43) provided efficient diagnostic sensitivity (71%) and specificity (83%) by using the production of tau levels and the ratio of A beta1-40 to A beta1-42(43), and an improvement in sensitivity (to 91%) was obtained in the longitudinal evaluation.

Aged↗
PubMed1998

Familial spinocerebellar ataxia with cerebellar atrophy, peripheral neuropathy, and elevated level of serum creatine kinase, gamma-globulin, and alpha-fetoprotein.

Here, we report a familial spinocerebellar ataxia (FSCA), which has clinical features similar to Friedreich's ataxia, an ataxia with isolated vitamin E deficiency, and ataxia telangiectasia. However, the serum levels of creatine kinase, gamma-globulin, and alpha-fetoprotein were elevated, and biochemical and genetic analyses ruled out diagnosis of these three ataxias as well as other FSCAs. Thus, this family is thought to have a new type of FSCA.

Adult↗
PubMed1998

Cerebral hypoperfusion and hypometabolism with altered striatal signal intensity in chorea-acanthocytosis: a combined PET and MRI study.

We studied cerebral perfusion and oxygen metabolism in three patients with chorea-acanthocytosis using positron-emission tomography and oxygen-15 labeled O2 and CO2. High-field magnetic resonance imaging also was performed. Regional cerebral blood flow (rCBF) and oxygen metabolism (rCMRO2) were significantly reduced in the caudate and putamen when compared with seven control subjects. Mild but significant reductions of rCBF (lower than the normal control values -2 SD) were found in the bilateral frontal, left temporal and parietal, and bilateral thalamic areas; rCMRO2 was reduced in the bilateral frontal and left temporal areas. Magnetic resonance imaging showed increased signal intensity accompanied by scattered bright spots in the caudate head and putamen on T2-weighted images; decreased signal intensity was shown at these sites on T1-weighted images. These findings were not observed in 10 neurologically normal volunteers and are rare in the common hyperkinetic form of Huntington's disease. Reduced cerebral perfusion and oxygen metabolism seem to be related to the intellectual and personality changes that occur in chorea-acanthocytosis. Combined positron-emission tomography and magnetic resonance imaging studies may improve diagnostic accuracy in patients with chorea-acanthocytosis and related disorders.

Acanthocytes↗
PubMed1998

In vivo 3D localized 13C spectroscopy using modified INEPT and DEPT.

The 3D localized 13C spectroscopy methods LINEPT and LODEPT, which are modifications of INEPT and DEPT, are proposed. As long as a 13C inversion pulse (180-degree pulse) is applied at 1/(4J) before the proton echo time in LINEPT and a 13C excitation pulse (90-degree pulse) is applied at 1/(2J) before the proton echo time in LODEPT, the proton echo time can be set to any value longer than 1/(2J) in LINEPT and longer than 1/J in LODEPT. As a result, the proton and the 13C pulses can be applied separately and these proton pulses can be made slice-selective pulses. These localization features of LINEPT and LODEPT were evaluated using a phantom consisting of a cylinder filled with ethanol placed inside another cylinder filled with oil, and localized ethanol spectra could be obtained. In vivo 3D localized 13C spectra from the brain of a monkey could be obtained using decoupled LINEPT, and glutamate C-4 appeared directly after the administration of glucose C-1, followed by the appearance of glutamate C-2, C-3 and glutamine C-2, C-3, C-4.

Animals↗
PubMed1998

Possible role of platelet-activating factor in the in vivo expression of tissue factor in neutrophils.

BACKGROUND: Recently, we demonstrated that neutrophils express tissue factor (TF) in a model of acute obstructive cholangitis (AOC). However, the regulation of TF expression was not clear. In this study, we clarified the role of platelet-activating factor (PAF) in TF expression in neutrophils. MATERIALS AND METHODS: In a model of AOC, intravenous PAF antagonist, (SM-12502, 200 mg/kg) was administered 5 min before sepsis was induced. Normal saline was given as a control. Coagulation parameters and TF activity were monitored for 6 h. Thereafter, the liver was harvested for histological examination. RESULTS: The percentage of neutrophils which stained positive for TF was significantly reduced by SM-12502 (74.9 +/- 19.3% vs 96.3 +/- 2.8%) (P < 0.01). The number of leukocytes infiltrating the liver was also significantly reduced. Coagulation abnormalities, TF activity, and focal necrosis of the hepatocytes were reduced by SM-12502. CONCLUSIONS: SM-12502 inhibits TF expression in neutrophils which have infiltrated the liver sinusoids, reducing the subsequent infiltration of leukocytes. These results suggest that PAF plays an important role in the expression of TF in neutrophils in vivo.

Animals↗
PubMed1998

Involvement of glutamic acid residue at position 7 in the formation of the intramolecular disulfide bond of Escherichia coli heat-stable enterotoxin Ip in vivo.

Escherichia coli heat-stable enterotoxin Ip (STIp) is a small peptide toxin composed of 18 amino acid residues containing three intramolecular disulfide bonds. We found previously that the bonds are formed by the catalysis of DsbA (a oxidoreductase) in periplasm [1]. To interact with DsbA, the STIp in periplasm must have a structure suitable as substrate. However, the amino acid residues contributing to the construction of this structure have not been elucidated. We mutated the codon for the glutamic acid at position 7 of STIp by oligonucleotide site-specific mutagenesis in vivo and analysed the STIp produced from the mutant gene. The intramolecular disulfide bonds were not formed in mutant STIp (Glu-7-->Ala), but were formed in mutant STIp (Glu-7-->Asp). Furthermore, we found that replacing the asparagine residue at position 11 and the proline residue at position 12 did not affect the disulfide bond formation of STIp. The results indicate that a negative charge at position 7 in the sequence of STIp is necessary for STIp to interact with DsbA in periplasm.

Amino Acid Sequence↗
PubMed1998

Need for TolC, an Escherichia coli outer membrane protein, in the secretion of heat-stable enterotoxin I across the outer membrane.

Escherichia coli heat-stable enterotoxin Ip (STIp) is a typical extracellular toxin consisting of 18 amino acid residues synthesized as a precursor of pre (amino acid residues 1 to 19), pro (amino acid residues 20 to 54), and mature (amino acid residues 55 to 72) regions. STIp synthesized in the cytoplasm must cross the inner and outer membranes to migrate into the extracellular environment. Previous studies showed that the precursor translocates across the inner membrane utilizing the general export pathway consisting of Sec proteins. However, it remains unclear how it crosses the outer membrane. In this study, we examined the effects of mutation of the tolC gene which encodes an E. coli outer membrane protein, TolC, on the release of STIp into the extracellular environment. The mutation reduced the amount of STIp released into culture supernatant and increased the amount of STIp accumulated in the periplasm. This indicates that TolC mediates the translocation of STIp across the outer membrane. The inability to transfer STIp in the periplasm into the culture supernatant was restored by introduction of the tolC gene into the mutant cells. In the mouse intestinal loop assay, living cells of the mutants did not show a positive response, but wild-type cells did. These results showed that TolC is involved in the translocation of STIp across the outer membrane.

Animals↗
PubMed1998

Clinical significance of reduced cerebral metabolism in multiple sclerosis: a combined PET and MRI study.

Magnetic resonance imaging (MRI) in patients with multiple sclerosis (MS) has provided major insights into the disease's natural history, and many studies have focussed on possible correlations between MRI findings and the clinical manifestations of MS. In contrast, there are few reports on possible relationships between functional imaging data and cognitive function. The present study assessed the relationship between clinical presentation and combined anatomical and functional imaging data in MS. Twenty patients with definite MS underwent MRI and positron emission tomography (PET) to evaluate cerebral blood flow (rCBF) and oxygen metabolism (rCMRO2). The relationships between these neuroimaging findings and clinical data, including the Expanded Disability Status Scale (EDSS), Mini-mental status scale, Hasegawa Dementia Scale and relapse time, were evaluated with Spearman's rank correlation coefficients. A general reduction in rCBF and rCMRO2 in the gray and white matter were found in the MS patients. EDSS was correlated with the number and size of the lesions on MRI and was negatively correlated with rCMRO2. A correlation between the decrease in rCMRO2 and the level of cognitive impairment was also found. The severity of cerebral hypometabolism was also related to the number of relapses. Morphological and functional findings obtained by MRI and PET are closely related to the clinical status in MS. Our results suggest that measurement of cerebral metabolism in MS has the potential to be an objective marker for monitoring disease activity and to provide prognostic information.

Adult↗
PubMed1998

[A case report of infective endocarditis caused by MRSA and characterized by pedicled vegetation on the posterior wall of left atrium].

We report here a case of active infective endocarditis caused by Methicilin-Resistant Staphylococcus aureus (MRSA). A 24-year-old woman was admitted to the Osaka Medical Collage Hospital with continuous fever. After admission, MRSA was detected by blood culture and chemotherapy with Vancomycin was started. However, after 1 week, her condition had not improved. Moreover, a pedicled vegetation on the posterior wall of the left atrium and mitral regurgitation due to prolapse of the anterior leaflet were revealed by transesophageal echocardiography. The vegetation grew to about 2 cm in diameter and prolapsed into the left ventricle during diastole. We performed an early operation although the infection was still active due to its rapid growth and the risk of embolism. There was a large pedicled vegetation on the posterior wall of the left atrium as shown by preoperative echocardiography, but the mitral valve appeared to be intact. Therefore, the vegetation was completely removed and the mitral annulus was plicated by Kay's method to treat the associated mitral regurgitation. Postoperatively, we administered VCM 2 g/day for 24 days. The course was uneventful. The patient was discharged from the hospital on the 31st postoperative day.

Adult↗