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Biomedical subjects

K Okamoto

Publications and source records attributed to K Okamoto.

At least 343 records · Page 19Linked to original sources
PubMed1999

Purely laparoscopic pylorus-preserving gastrectomy with extraperigastric lymphadenectomy for early gastric cancer: a case and technical report.

For the purpose of prevention of postgastrectomy syndrome and a less invasive and yet curative oncological resection, a purely laparoscopic pylorus-preserving gastrectomy with extraperigastric lymphadenectomy was performed for a patient with early gastric cancer located in the middle third of the stomach. The patient's postoperative course was uneventful. During his postoperative recovery, the patient experienced very little pain and used analgesic medication only one time. This operation appeared to be oncologically adequate. As of the seventh postoperative month, the patient never experienced dumping syndrome or alkaline reflux gastritis. This procedure is technically feasible and an excellent option because of its reduced surgical invasiveness and better postoperative quality of life.

Adenocarcinoma↗
PubMed1998

Molecular analysis of a de novo mutation for spinocerebellar ataxia type 6 and (CAG)n repeat units in normal elder controls.

Spinocerebellar ataxia type 6 (SCA6) is an autosomal dominant spinocerebellar degenerative disease caused by CAG repeat expansions in the human alpha1A voltage-dependent calcium channel subunit gene (CACNL1A4). We analyzed 15 SCA6 patients in 14 unrelated Japanese families and 52 healthy Japanese aged over 74 years. Sequence analysis was performed to determine the correct number of CAG repeats. The expanded CAG repeat number was 23.6+/-2.1 (mean+/-S.D., n=15) with a range of 20-29, and the shortest expanded allele was 20 repeats. Moreover, the analysis of normal subjects revealed that the CAG repeat number of normal alleles was 12.3+/-1.9 (n=104) with a range of 7-18. We concluded that the normal range of CAG repeats in the CACNL1A4 gene is 18 or less, and that the disease range is 20 or more. Of 15 SCA6 patients, three sporadic cases were observed. In one male patient with 26 CAG repeats, the CAG repeat numbers of his parents were within normal range. His expanded allele was considered to be caused by an expansion of a normal allele from his mother (14 or 17 repeats). This is the first SCA6 case which was genetically proven to occur due to a de novo mechanism, suggesting that larger CAG repeats of normal alleles in the CACNL1A4 gene may be unstable and result in full expansion.

Aged↗
PubMed1998

Signal transduction in hypoxic cells: inducible nuclear translocation and recruitment of the CBP/p300 coactivator by the hypoxia-inducible factor-1alpha.

In response to decreased cellular oxygen concentrations the basic helix-loop-helix (bHLH)/PAS (Per, Arnt, Sim) hypoxia-inducible transcription factor, HIF-1alpha, mediates activation of networks of target genes involved in angiogenesis, erythropoiesis and glycolysis. Here we demonstrate that the mechanism of activation of HIF-1alpha is a multi-step process which includes hypoxia-dependent nuclear import and activation (derepression) of the transactivation domain, resulting in recruitment of the CREB-binding protein (CBP)/p300 coactivator. Inducible nuclear accumulation was shown to be dependent on a nuclear localization signal (NLS) within the C-terminal end of HIF-1alpha which also harbors the hypoxia-inducible transactivation domain. Nuclear import of HIF-1alpha was inhibited by either deletion or a single amino acid substitution within the NLS sequence motif and, within the context of the full-length protein, these mutations also resulted in inhibition of the transactivation activity of HIF-1alpha and recruitment of CBP. However, nuclear localization per se was not sufficient for transcriptional activation, since fusion of HIF-1alpha to the heterologous GAL4 DNA-binding domain generated a protein which showed constitutive nuclear localization but required hypoxic stimuli for function as a CBP-dependent transcription factor. Thus, hypoxia-inducible nuclear import and transactivation by recruitment of CBP can be functionally separated from one another and play critical roles in signal transduction by HIF-1alpha.

Amino Acid Substitution↗
PubMed1998

Arg-gingipain acts as a major processing enzyme for various cell surface proteins in Porphyromonas gingivalis.

Arg-gingipain (RGP) is an Arg-X-specific cysteine proteinase produced by the Gram-negative anaerobe Porphyromonas gingivalis and has been shown to be a potent virulence factor in progressive periodontal disease (Nakayama, K., Kadowaki, T., Okamoto, K., and Yamamoto, K. (1995) J. Biol. Chem. 270, 23619-23626). In this study, we provide evidence that RGP acts as a major processing enzyme for various cell surface and secretory proteins in P. gingivalis. Fimbrilin, a major component of fimbriae, remained in the precursor form in the RGP-null mutant. Prefimbrilin expressed in Escherichia coli was converted to the mature fimbrilin in vitro when incubated with purified RGP, but its conversion was suppressed by potent RGP inhibitors. The results were consistent with the electron microscopic observation indicating little or no fimbriation in the RGP-null mutant. The immunogenic 75-kDa cell surface protein was also shown to retain its proform in the RGP-null mutant. In addition, Lys-gingipain (KGP) was found to be abnormally processed in the RGP-null mutant. In contrast, both prefimbrilin and the 75-kDa protein precursor were processed to their respective mature forms in the KGP-null mutant, suggesting that KGP is not involved in the normal processing mechanisms of these proteins. These results suggest that RGP not only acts as a direct virulence factor but also makes a significant contribution as a major processing enzyme to the virulence of P. gingivalis.

Adhesins, Bacterial↗
PubMed1998

Pituitary adenylate cyclase-activating polypeptide (PACAP) induces duodenal phasic contractions via the vagal cholinergic nerves in sheep.

This study examined the effect of intravenous infusion of pituitary adenylate cyclase-activating polypeptide (PACAP) on duodenal motility in sheep and the mechanism of the action of PACAP. The bilateral cervical vagus nerves were coiled with a cooling device under anesthesia. Duodenal motility was recorded by manometry in conscious animals. PACAP-27, PACAP-38 and vasoactive intestinal polypeptide (VIP) were infused intravenously at 3, 10, 30 and 100 pmol/kg in phase II of the duodenal migrating motor complexes (MMC). PACAP-27 induced a cluster of phasic contractions of the duodenum, while PACAP-38 only augmented the spontaneous contractions. The pattern of PACAP-27-induced contractions was different from that of phase III of MMC, and the contractions were followed by a quiescence period. VIP at only the highest dose induced phase III-like activity of the duodenum. Both the intravenous background infusion of atropine at 10 nmol/kg/min and the reversible cooling blockade of the bilateral cervical vagus nerves blocked the effect of PACAP-27. The application of PACAP-27 at concentrations ranging from 10 nM to 1 microM did not induce contractions of ovine duodenal smooth muscles. These results indicate that PACAP contracts the ovine duodenum via the vagal cholinergic efferent fibers, suggesting that PACAP acts on the central nervous system.

Animals↗
PubMed1998

Involvement of a lysine-specific cysteine proteinase in hemoglobin adsorption and heme accumulation by Porphyromonas gingivalis.

The oral anaerobic bacterium Porphyromonas gingivalis, a major pathogen of advanced adult periodontitis, produces a novel class of cysteine proteinases in both cell-associated and secretory forms. A lysine-specific cysteine proteinase (Lys-gingipain, KGP), as well as an arginine-specific cysteine proteinase (Arg-gingipain), is a major trypsin-like proteinase of the organism. Recent studies indicate that the secreted KGP is implicated in the destruction of periodontal tissue and the disruption of host defense mechanisms. In this study, we have constructed a KGP-deficient mutant to determine whether the cell-associated KGP is important for pathophysiology of the organism. Although the mutant retained the strong ability to disrupt the bactericidal activity of polymorphonuclear leukocytes, its hemagglutination activity was reduced to about one-half that observed with the wild-type strain. More important, the mutant did not form black-pigmented colonies on blood agar plates, indicating the defect of hemoglobin adsorption and heme accumulation. Immunoblot analysis showed that the expression of a 19-kDa hemoglobin receptor protein, which is thought to be responsible for hemoglobin binding by the organism, was greatly retarded in this mutant. The mutant also showed a marked decrease in the ability to degrade fibrinogen. These results suggest the possible involvement of KGP in the hemoglobin binding and heme accumulation of the organism and in the bleeding tendency in periodontal pockets.

Adhesins, Bacterial↗
PubMed1998

The sorting of mitochondrial DNA and mitochondrial proteins in zygotes: preferential transmission of mitochondrial DNA to the medial bud.

Green fluorescent protein (GFP) was used to tag proteins of the mitochondrial matrix, inner, and outer membranes to examine their sorting patterns relative to mtDNA in zygotes of synchronously mated yeast cells in rho+ x rho0 crosses. When transiently expressed in one of the haploid parents, each of the marker proteins distributes throughout the fused mitochondrial reticulum of the zygote before equilibration of mtDNA, although the membrane markers equilibrate slower than the matrix marker. A GFP-tagged form of Abf2p, a mtDNA binding protein required for faithful transmission of rho+ mtDNA in vegetatively growing cells, colocalizes with mtDNA in situ. In zygotes of a rho+ x rho+ cross, in which there is little mixing of parental mtDNAs, Abf2p-GFP prelabeled in one parent rapidly equilibrates to most or all of the mtDNA, showing that the mtDNA compartment is accessible to exchange of proteins. In rho+ x rho0 crosses, mtDNA is preferentially transmitted to the medial diploid bud, whereas mitochondrial GFP marker proteins distribute throughout the zygote and the bud. In zygotes lacking Abf2p, mtDNA sorting is delayed and preferential sorting is reduced. These findings argue for the existence of a segregation apparatus that directs mtDNA to the emerging bud.

Crosses, Genetic↗
PubMed1998

Functional replacement of OprJ by OprM in the MexCD-OprJ multidrug efflux system of Pseudomonas aeruginosa.

For characterization of the MexCD-OprJ efflux system of Pseudomonas aeruginosa involved in resistance to fluoroquinolones and the fourth-generation cephems, we constructed mexC, mexD or oprJ mutants from the nfxB-type PAO strains by insertion mutagenesis. The gene products in the resultant mutants were examined by immunoblot assay using murine and rabbit antibodies developed against purified protein and synthetic oligopeptides. Susceptibility of the mexC (MexC- MexD- OprJ-) and mexD (MexC+ MexD- OprJ-) mutants to fluoroquinolone and the fourth-generation cephems was comparable to that of the wild-type strain PAO1. However, the oprJ mutant (MexC+ MexD+ OprJ-) was still less susceptible than PAO1, since a MexCD-OprM chimera system, which generated from a functional assist of the constitutively expressed OprM, can function in the efflux of the antimicrobial agents in the oprJ mutant. In fact, transformation of the oprJ mutant with an OprM-expression plasmid decreased the former's susceptibility to the levels exhibited by the nfxB mutant without affecting the substrate specificity of MexCD-OprJ.

Animals↗
PubMed1998

Restoration of the glucocorticoid receptor function by the phosphodiester compound of vitamins C and E, EPC-K1 (L-ascorbic acid 2-[3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-1-benzopyran-6-yl hydrogen phosphate] potassium salt), via a redox-dependent mechanism.

We examined the effect of the novel antioxidant EPC-K1 (L-ascorbic acid 2-[3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H -1-benzopyran-6-yl hydrogen phosphate] potassium salt) on glucocorticoid receptor function. We used cloned CHOpMTGR cells in which human glucocorticoid receptor cDNA was stably transfected and the glucocorticoid receptor was expressed at high levels. We recently suggested that glucocorticoid-mediated gene expression is modulated via the cellular redox state [Makino et al., J Clin Invest 98: 2469-2477, 1996]. In the present study, this issue was clearly evidenced by the finding that cellular treatment with H2O2 decreased the ligand binding and transcriptional activity of the glucocorticoid receptor, and we showed that these inhibitory effects of H2O2 were effectively titrated by the addition of EPC-K1. Moreover, DNA-binding activity of the bacterially expressed DNA-binding domain of the glucocorticoid receptor was repressed by the thiol-oxidizing reagent diamide; EPC-K1 also counteracted this repressive effect of diamide. Thus, the redox state was indicated to influence glucocorticoid receptor function at various steps, and EPC-K1 may be useful in restoring the cellular glucocorticoid-responsiveness in oxidative conditions.

Animals↗
PubMed1998

A Japanese family with adrenoleukodystrophy with a codon 291 deletion: a clinical, biochemical, pathological, and genetic report.

We report a Japanese family with adrenoleukodystrophy (ALD) with a three base pair deletion (delGAG 291) in the ALD gene. A variety of phenotypes were observed within this family. While the proband (patient 1) was classified as having a rare intermediate type of adult cerebral and cerebello-brain stem forms, his younger brother (patient 2) and nephew (patient 3) had a childhood ALD type. Another nephew (patient 4) of patient 1 was classified as having an adolescent form. The tau level in the cerebrospinal fluid (CSF) in patient 1 was as high as that of patients with Alzheimer's disease (AD). His brain magnetic resonance image (MRI) showed abnormalities in the bilateral cerebellar hemispheres and brain stem, but not in the cerebral white matter, where marked reductions of the cerebral blood flow and oxygen metabolism were clearly demonstrated by positron emission tomography (PET). In patients 2 and 3, the autopsy findings showed massive demyelination of the cerebral white matter with sparing of the U-fibers, compatible with the findings of childhood ALD. Oleic and erucic acids (Lorenzo's Oil) were administered to patients 1 and 4, but sufficient effectiveness was not obtained. The findings in this family suggest that delGAG291 is part of the cause of Japanese ALD with phenotypic variations. Moreover, although the scale of the study is limited, there is a possibility that PET can detect an insidious lesion which is undetectable by computed tomogram (CT) or MRI analysis, and that the higher level of tau reflects the process of neuronal degeneration in ALD. Lorenzo's Oil should be given in the early stage.

Adrenoleukodystrophy↗
PubMed1998

Combination assay of CSF tau, A beta 1-40 and A beta 1-42(43) as a biochemical marker of Alzheimer's disease.

Cerebrospinal fluid samples from a total of 157 subjects consisting of 55 patients with sporadic Alzheimer's disease (AD), 34 normal controls, 23 patients with non-AD dementia, and 45 with other neurological diseases were examined by ELISA of tau, A beta 1-40, and A beta 1-42(43). The AD group had a significantly higher level of tau than the normal control group (P < 0.001), and the diagnostic sensitivity was 31% and specificity was 94%. CSF A beta 1-40 levels did not show any significant differences. Although the level of A beta 1-42(43) was decreased significantly in the AD group compared to the control group (P < 0.005), the overlap of A beta 1-42(43) levels among all groups meant that none of the AD samples exceeded the cut-off value, the mean 2SD of normal control subjects. Reduction of A beta 1-42(43) levels in AD resulted in a significant increase in the ratio of A beta 1-40 to A beta 1-42(43) (A beta ratio) as an improved marker. The diagnostic sensitivity and specificity of A beta ratio were 51% and 82% respectively. The three indexes, using the tau level and A beta ratio (tau or A beta ratio, deviation score and tau x A beta ratio), showed better sensitivity (58%, 67%, 69%) and specificity (82%, 86%, 88%) than previously reported methods. Combination assay for CSF tau, A beta 1-40 and A beta 1-42(43) in CSF is a biological marker of AD and may be useful to biochemically monitor subjects under treatment.

Adult↗
PubMed1998

Activated microglia cause iron-dependent lipid peroxidation in the presence of ferritin.

Ferritin contains most of the iron found in the brain, and the release of iron from ferritin has an essential role in iron-dependent lipid peroxidation. We examined the effect of cultured microglia on the ferritin-dependent lipid peroxidation of phospholipid liposomes monitored by the formation of thiobarbituric acid-reactive substances. Microglia stimulated by phorbol myristate acetate caused lipid peroxidation in the presence of ferritin. This lipid peroxidation was mediated by superoxide produced by the microglia and iron released from the ferritin. Lipid peroxidation induced by activated microglia may be partly responsible for the oxidative damage that is thought to occur in Parkinson's disease and other neurodegenerative disorders.

Animals↗
PubMed1998

Inhibition of mesangial cell proliferation by E2F decoy oligodeoxynucleotide in vitro and in vivo.

The transcription factor E2F coordinately activates several cell cycle-regulatory genes. We attempted to inhibit the proliferation of mesangial cells in vitro and in vivo by inhibiting E2F activity using a 25-bp decoy oligodeoxynucleotide that contained consensus E2F binding site sequence (E2F-decoy) as a competitive inhibitor. The decoy's effect on human mesangial cell proliferation was evaluated by [3H]thymidine incorporation. The E2F decoy inhibited proliferation in a concentration-dependent manner, whereas a mismatch control oligodeoxynucleotide had little effect. Electrophoretic mobility shift assays demonstrated that the decoy's inhibitory effect was due to the binding of the decoy oligodeoxynucleotide to E2F. The effect of the E2F decoy was then tested in a rat anti-Thy 1.1 glomerulonephritis model. The E2F decoy oligodeoxynucleotide was introduced into the left kidney 36 h after the induction of glomerulonephritis. The administration of E2F decoy suppressed the proliferation of mesangial cells by 71%. Furthermore, treatment with the E2F decoy inhibited the glomerular expression of proliferating cell nuclear antigen at the protein level as well as the mRNA level. These findings indicate that decoy oligonucleotides can suppress the activity of the transcription factor E2F, and may thus have a potential in treating glomerulonephritis.

Animals↗
PubMed1998

Antioxidant properties of nicergoline; inhibition of brain auto-oxidation and superoxide production of neutrophils in rats.

Oxidative stress has been suggested to adversely influence cerebrovascular disorders and some neurodegenerative disorders. We examined whether nicergoline, an agent widely used for treating cerebrovascular disorders and senile mental impairment, possesses antioxidant activities and some beneficial effect on neutrophils generating free radicals. Although nicergoline did not scavenge superoxide produced from a superoxide-generating system, it significantly inhibited superoxide secretion from stimulated neutrophils. Auto-oxidation of brain homogenate of rats, monitored by formation of thiobarbituric acid-reactive substances, was suppressed by nicergoline in a dose-dependent manner. The oxidation of the homogenate was accelerated by activated neutrophils and was significantly suppressed by nicergoline. These observations suggest that nicergoine is an antioxidant that inhibits not only lipid peroxidation but also free radical generation from neutrophils. These properties of nicergoline should be beneficial in some pathological conditions including cerebrovascular and neurodegenerative disorders in which oxidative stress may have a pathoetiological role.

Animals↗
PubMed1998

Inhibitory effects of plumbagin and juglone on azoxymethane-induced intestinal carcinogenesis in rats.

The effects of two naphthoquinones, juglone and plumbagin, and an isocoumarin, hydrangenol, on intestinal carcinogenesis in rats were examined by dietary exposure during the initiation phase. Starting at 5 weeks of age, male F344 rats were fed the diets containing either of the test chemicals at a concentration of 200 ppm or the control diet without the compounds. At 6 weeks of age, all animals were treated with s.c. injections of azoxymethane (AOM) (15 mg/kg body weight, once weekly for 3 weeks) or saline alone. Animals fed experimental diets were changed to the control diet 1 week after the last carcinogen treatment. Animals given plumbagin together with the carcinogen had a lower incidence (41%) and smaller multiplicity (0.48 +/- 0.62) of tumors in the entire intestine compared with those exposed to carcinogen alone (68% and 1.04 +/- 0.62) (P < 0.05 and < 0.01, respectively). The incidence and multiplicity of tumors in the small intestine (7% and 0.07 +/- 0.25) and the multiplicity of tumors in the entire intestine (0.60 +/- 0.76) of animals treated with juglone and the carcinogen were significantly less than those of animals treated with carcinogen alone (P < 0.05 in each). Hydrangenol tended to decrease the incidence and the multiplicity of tumors in the entire intestine induced by AOM, but the effect was not statistically significant. The present data suggest that the naphthoquinones, juglone and plumbagin, could be promising chemopreventive agents for human intestinal neoplasia.

Animals↗
PubMed1998

Enhancement of expression of stress proteins by agents that lower the levels of glutathione in cells.

The effects of diethyl maleate and buthionine sulfoximine, agents that lower cellular levels of glutathione, on expression of hsp27 and alphaB crystallin in response to stress were studied. When C6 rat glioma cells were treated with 100 microM arsenite for 1 h, accumulation of the two proteins, estimated by specific immunoassays, was markedly enhanced by additional exposure to 1 mM diethyl maleate or 2.5 mM buthionine sulfoximine. The latter also increased heat- and CdCl2-induced accumulation of hsp27 and alphaB crystallin. Stress-induced accumulation of hsp70, estimated by Western blotting analysis, was also enhanced by these agents. Northern blotting analysis revealed increase in levels of mRNAs for hsp27, alphaB crystallin and hsp70. The period of heat shock element (HSE)-binding activity of heat shock factor (HSF) stimulated by arsenite was extended by addition of diethyl maleate and buthionine sulfoximine. The induced phosphorylated state of HSF1 was also prolonged by diethyl maleate. Although exposure of cells to diethyl maleate alone for 1 h caused neither accumulation of hsp27, alphaB crystallin and hsp70 nor expression of mRNAs for these proteins, HSE-binding activity of HSF was stimulated. However, the activated HSF was not phosphorylated. These results suggest that diethyl maleate induces an intermediate state of HSF that binds to HSE but is transcriptionally inert. The mechanism is unclear but the levels of glutathione in cells that were exposed to diethyl maleate or buthionine sulfoximine were markedly decreased.

Animals↗
PubMed1998

Changes in expression of the antigen recognized by monoclonal antibody A7 in human pancreatic carcinoma cells following exposure to anticancer agents.

Techniques which can increase the expression level of tumor-associated antigens may improve immunotargeting therapy. We studied the reactivity of MAb A7 toward an antigen expressed on the surface of the human pancreatic cancer cell line HPC-YS after treatment with various antitumoral agents. When we applied 1 microg/ml mitomycin C (MMC) or 0.1 microg/ml neocarzinostatin (NCS) for 1 h, A7 recognizing antigen expression was enhanced until 24 h after the treatments. At a dose that completely suppressed cell growth, increased antigen expression was maintained for 96 h. Therefore, this study suggests that the combined application of an anticancer drug and MAb A7 may be useful for immunotargeting chemotherapy.

Antibiotics, Antineoplastic↗