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Biomedical subjects

H Aoki

Publications and source records attributed to H Aoki.

At least 109 records · Page 6Linked to original sources

A study of orientation in a zero gravity environment by means of virtual reality simulation.

When the International Space Station (ISS) is completed and starts its operation, crew members will be stationed for three months or more in orbit aboard the ISS. As they stay longer in the space environment, "habitability" for them will become most important in the design of the interior space. One of the problems about habitability in a zero gravity (0 G) environment is disorientation. Crew members have difficulty in discriminating between "up" and "down" and more serious disorientations may cause space motion sickness. Crew members rely on visual perception to orient themselves because they can't use their sense of equilibrium in a 0 G environment. Although color and the direction of equipment of Space Shuttles or modules has been considered, no systematic study has been conducted on interior space. This study intended to clarify how people acquire visual information and recognize their orientation in a 0 G environment by an experiment in which a subject wears a head-mounted display (HMD) and enters a virtual weightless state represented by computer graphics (CG). Visual information of a room and the degree-of-freedom of motion were varied to examine the influence of the conditions on such a simple task as movement through several connected modules, and the performance and the behavior of each subject were investigated.

Humans↗

Induction of antibodies in mice by a recombinant baculovirus expressing pseudorabies virus glycoprotein B in mammalian cells.

The glycoprotein gB of pseudorabies virus (PrV) was expressed in various mammalian cells by a recombinant baculovirus carrying the PrV gB gene under the control of the CAG promoter. When the recombinant baculovirus was inoculated into the stable porcine kidney cell line CPK, expression of PrV gB was detected by immunofluorescent antibody analysis and a 155 kDa of protein, which has the same molecular mass as the native PrV gB, was detected by Western blotting. High levels of expression of PrV gB were observed in BHK-21, HmLu-1 and SK-H cell lines. Furthermore, anti-PrV gB-specific antibodies against PrV gB protein were detected by the enzyme-linked immunosorbent assay in mice inoculated the recombinant baculovirus. The recombinant baculovirus containing the PrV glycoprotein gB gene under the CAG promoter could be a candidate for a pseudorabies vaccine.

Animals↗

Association between high serum alanine aminotransferase levels and more rapid development and higher rate of incidence of hepatocellular carcinoma in patients with hepatitis C virus-associated cirrhosis.

BACKGROUND: Many studies have demonstrated in animal experiments that persistent inflammation may accelerate the development of carcinoma. In this article, the question of whether the persistent elevation of serum alanine aminotransferase (ALT) levels (which represents the inflammatory necrosis of hepatocytes) correlates with the development of hepatocellular carcinoma (HCC) was studied in patients with early stage hepatitis C virus (HCV)-associated cirrhosis. METHODS: Sixty-nine consecutive patients with biopsy proven HCV-associated cirrhosis (mostly Child's Stage A) who had been followed for >5 years for the development of HCC were studied. They were subdivided into 3 groups according to their serum ALT levels: Group A was comprised of 28 patients whose annual average serum ALT level was persistently high (>/= 80 IU) (high ALT group), Group B was comprised of 28 patients whose annual average serum ALT level was persistently low (< 80 IU) (low ALT group), and Group C was comprised of 13 unclassified patients. The patients had been studied prospectively with frequent ultrasonography and magnetic resonance imaging or computed tomography for > 5 years. RESULTS: In the high ALT group HCC developed in 71.4% of patients compared with 25.0% in the low ALT group over the observation period (P < 0.005). The 5-year rate of incidence of HCC in the high ALT group was as high as 53.6% compared with only 7.1% in the low ALT group (P < 0.001). The expected interval between the diagnosis of cirrhosis and the development of HCC was 6.0 +/- 0.7 years (mean +/- standard error) in the high ALT group and 12.7 +/- 1.2 years in the low ALT group (P < 0.001). CONCLUSIONS: The results of the current study demonstrated that the development of HCC was more rapid in the high ALT group with HCV-associated cirrhosis.

Aged↗

Effect of Na(+) concentration on the subgel phases of negatively charged phosphatidylglycerol.

The effect of Na(+) concentration on the subgel phase of dimyristoylphosphatidylglycerol (DMPG) was investigated by differential scanning calorimetry (DSC) and negative stain electron microscopy, and the results were compared with dipalmitoylphosphatidylglycerol (DPPG). The conversion mode of DMPG vesicle to the subgel phase by annealing at 5 degrees C was grouped into two types depending on whether Na(+) concentration is above or below 200-250 mM. For [Na(+)]>200-250 mM, the subgel phase of a crystalline superstructure of bilayers wrapped in a cylinder was attained during a 24-h period of annealing and transformed directly to the liquid crystal phase on heating. For [Na(+)]<200-250 mM, two subgel phases which transform to the gel phase on heating were observed after annealing up to 24 h. Both subgel phases showed belt-like structures composed of loosely and closely stacked lamellae, respectively, and their fractions were found to depend on Na(+) concentration. With a further annealing up to 30 days, only the closely stacked subgel phase converted subsequently into the cylindrical superstructure of a more ordered phase. Similar two subgel phases were detected for DPPG at [Na(+)]< or =100 mM. The difference in the relative enthalpy between the gel and subgel phases was investigated from the van der Waals interaction energy between the hydrocarbon chains.

Journal Article↗

Host expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in normal colon tissue affects metastatic potential of colorectal cancer.

PURPOSE: To clarify the mechanism of cancer cell invasion, we paid close attention to the role of matrix metalloproteinases and tissue inhibitors of metalloproteinases in normal tissue that is located in the same organ as the cancer. METHODS: Samples were obtained from a tumor lesion and normal tissue in the resected large intestine of 59 patients with colorectal cancer, including 13 cases with liver metastasis (Group A) and 46 cases without liver metastasis (Group B). In each sample the expression of m-RNA for matrix metalloproteinase-2, matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, and tissue inhibitor of metalloproteinase-2 was examined using reverse transcription-coupled polymerase chain reaction and southern hybridization. RESULTS: In normal colon tissue the expression rate of matrix metalloproteinase-2 in Group A (76.9 percent) was significantly higher than that of Group B (15.2 percent; P < 0.0001). Regarding the expression pattern of m-RNA of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in normal colon tissue, Group B included 24 cases with matrix metalloproteinase-2 negative, and tissue inhibitor of metalloproteinase-2 positive (24/46; 52.2 percent). Conversely, Group A had only one case with matrix metalloproteinase-2 negative and tissue inhibitor of metalloproteinase-2 positive (1/13; 7.7 percent; P = 0.0107). In addition, the ratio of cases with matrix metalloproteinase-2 positive and tissue inhibitor of metalloproteinase-2 negative in Group A was 30.8 percent (4/13), which was a significantly higher rate than that in Group B (3/46; 6.5 percent; P = 0.0170). CONCLUSION: We think that the expression pattern of m-RNA of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in normal colon tissue is closely related to liver metastasis in colon cancer patients.

Aged↗

Identification of a ribosomal ATPase in Escherichia coli cells.

Eukaryotic ribosomes harbor an ATPase activity that has been shown to be essential for translation elongation in some lower fungi. Here we report the first identification of a ribosome bound ATPase, RbbA, in E. coli cells. RbbA accounts for most of the ATPase activity associated with 70S ribosomes and 30S ribosomal subunits. Both native and recombinant RbbA were purified and shown to possess ribosome-dependent ATPase activities and to stimulate polyphenylalanine synthesis in vitro. Biochemically, RbbA is similar to the fungi-specific translation elongation factor 3 (EF-3) and cross-reacts with antibody raised against EF-3. The gene encoding RbbA is identified as ORF yhih and the predicted RbbA amino acid sequence is 40% similar to that of the C-terminal half of EF-3. The discovery of a ribosomal ATPase in a prokaryotic cell suggests a common, conserved function for these proteins in translation.

Adenosine Triphosphatases↗

Molecular characterization of a prokaryotic translation factor homologous to the eukaryotic initiation factor eIF4A.

Initiation of translation involves a complex series of reactions that result in the formation of an initiation complex at the proper start site of the mRNA. These reactions, particularly those that involve the binding of the mRNA to the small subunit of the ribosome, are not fully understood. Here we show that one of the factors (W2) required to reconstitute translation in E. coli is encoded by the deaD gene which harbors 87% amino acid sequence similarly to the eukaryotic (eIF4A). Antibodies against the eukaryotic eIF4A cross-react with the E. coli protein. We describe the overexpression of the W2 protein from recombinant clones and its purification in one step by the use of a His tag at the N-terminus of its sequence. We report a rapid assay for the W2 protein that scores for initiation and elongation programmed by a native mRNA template. The W2 protein promotes initiation programmed by the mRNA that harbors secondary structures. The W2 protein is not required in standard initiation assays programmed by synthetic mRNAs of defined sequence that lack this feature. We conclude that W2 is an important factor for initiation in eukaryotic and prokaryotic cells.

Adenosine Triphosphate↗

Two novel lanostane-type triterpenes from the stem bark of abies mariesii

Two novel lanostane-type triterpene lactones, 3alpha, 11alpha-dihydroxy-7-oxolanosta-8,24-dien-26,23(R)-olide (1) and 1alpha,3alpha,11alpha-trihydroxy-7-oxolanosta-8,24-dien-26, 23(R)-olide (2), were isolated from the stem bark of Abies mariesii, together with a known triterpene, 3-oxo-9betaH-lanosta-7,24-dien-26, 23(R)-olide. The structures of 1 and 2 were determined on the basis of 2D NMR techniques.

Journal Article↗

A newly developed assay for melatonin using cells expressing human mel-1a receptor.

We have developed a radioreceptor binding assay (RRA) method for melatonin using membranes from Chinese hamster ovary cells that can stably express human mel-1a receptors. We measured melatonin levels in plasma samples collected every 4h for 24h using the RRA and radioimmunoassay (RIA) methods, simultaneously. There was a statistically significant correlation between the melatonin levels measured by the two methods, this newly developed method providing a sensitive bioassay. As it is possible to circumvent the cross-reactivity usually occurring in the RIA method, this method may be an important tool for detecting bioactive substances relative to the mel-1a receptor.

Animals↗

erbB-2 overexpression but no activation of beta-Catenin gene in extramammary Paget's disease.

Our previous study in extramammary Paget's disease showed neither p53 mutations nor allelic loss at selected loci implicated in other cancers, suggesting a pathogenesis of this skin cancer different from other common epithelial malignancies. To examine further the genetic defects in extramammary Paget's disease, we carried out molecular genetic analyses in 31 tumor samples obtained from 27 cases of extramammary Paget's disease without underlying malignancies. Immunohistochemistry using CB-11 monoclonal antibody revealed either membrane or cytoplasmic erbB-2 oncoprotein overexpression in none of the 13 primary in situ tumors, but in one recurrent in situ tumor, 10 of 13 invasive primary tumors and two of four lymph node metastases. Sensitive dual color fluorescence in situ hybridization analysis using probes for erbB-2 gene locus and chromosome 17 pericentromere, however, revealed different erbB-2 gene status in the erbB-2 overexpressing tumors. One recurrent in situ tumor and one lymph node metastasis showed definite gene amplification characterized by multiple scattered signals or a few large clustered erbB-2 signals, whereas four tumors with predominantly cytoplasmic erbB-2 overexpression were thought to have low-grade gene amplification. The remaining six tumors overexpressing erbB-2 showed no increase of erbB-2 copy numbers. No evidence of abnormal activation of the beta-catenin gene, a critical mediator of Wnt signaling pathway, in any tumor by immunohistochemical staining and by direct sequencing and reverse transcription-polymerase chain reaction analysis was found. Frequent overexpression of erbB-2 by either gene amplification or possible transcriptional activation in invasive primary tumors and metastases suggests an important part for this oncogene in the progression of extramammary Paget's disease.

Aged↗

Roles of bone morphogenetic protein type I receptors and Smad proteins in osteoblast and chondroblast differentiation.

The biological effects of type I serine/threonine kinase receptors and Smad proteins were examined using an adenovirus-based vector system. Constitutively active forms of bone morphogenetic protein (BMP) type I receptors (BMPR-IA and BMPR-IB; BMPR-I group) and those of activin receptor-like kinase (ALK)-1 and ALK-2 (ALK-1 group) induced alkaline phosphatase activity in C2C12 cells. Receptor-regulated Smads (R-Smads) that act in the BMP pathways, such as Smad1 and Smad5, also induced the alkaline phosphatase activity in C2C12 cells. BMP-6 dramatically enhanced alkaline phosphatase activity induced by Smad1 or Smad5, probably because of the nuclear translocation of R-Smads triggered by the ligand. Inhibitory Smads, i.e., Smad6 and Smad7, repressed the alkaline phosphatase activity induced by BMP-6 or the type I receptors. Chondrogenic differentiation of ATDC5 cells was induced by the receptors of the BMPR-I group but not by those of the ALK-1 group. However, kinase-inactive forms of the receptors of the ALK-1 and BMPR-I groups blocked chondrogenic differentiation. Although R-Smads failed to induce cartilage nodule formation, inhibitory Smads blocked it. Osteoblast differentiation induced by BMPs is thus mediated mainly via the Smad-signaling pathway, whereas chondrogenic differentiation may be transmitted by Smad-dependent and independent pathways.

Activin Receptors↗

Hepatitis C virus and hepatocarcinogenesis.

Although human hepatocellular carcinoma (HCC) is one of the most common types of tumors in the world, the molecular mechanisms underlying hepatitis-C-related human hepatocarcinogenesis are still not clear. HCC is accompanied by virus infections in most cases, and it is suggested that hepatitis B virus and hepatitis C virus (HCV) significantly influence the oncogenic process. The persistence of inflammation following HCV infection is reportedly related to carcinogenesis, and the mechanism of chronic inflammation has been approached by taking viral, immunologic, cytokine and apoptotic responses into consideration. With the progress made in molecular biology, the functional abnormality of oncogenes/tumor suppressor genes has been identified and, apart from the p53 gene, involvement of the IGF-II gene has also been described recently. Furthermore, it has been suggested that uncontrolled proliferation of cancer cells might be based on abnormal regulation of intracellular signal transduction pathways. Here we review the cutting edge of molecular hepatitis C virology in terms of virus-cell interactions, which may contribute to the development of human HCCs. We also discuss the recent progress made in the molecular and cell biology of human hepatocarcinogenesis.

Chromosome Aberrations↗

[Double cancer observed from bladder cancer].

BACKGROUND: In recent years, despite of the improvement of treatment results for cancer and long life, the occurrence of second primary cancer was increased. In this paper, we analyzed present condition of double cancer observed with bladder cancer in our hospital. METHOD: Last 21 years, we have treated 969 cases (828 male and 141 female) of primary bladder cancer. For those cases, we analyzed in term of frequency, involved organ, age, interval between two cancer occurrence, risk factor and prognosis of double cancer patients. RESULT: Of 969 cases with bladder cancer, 81 cases (8.36%) had double cancer involving 6 cases (0.61%) of triple cancer. In sex, 70 males (9.78%) and 11 females (7.80%) had double cancer. As involved organs, 25 cases (3.02%) had in prostate, 23 cases (2.37%) in stomach, 3 case (2.13%) in breast, 14 cases (1.44%) in colon and rectum. In diagnosis timing of complicated cancer from bladder cancer, 28 cases (34.6%) were diagnosed previously to bladder, 28 cases (34.6%) were simultaneously and 31 cases (38.3%) were secondary. An average interval of diagnosis of two cancer were 49 +/- 42.5 months. An average age of occurrence of second cancer was 70.3 +/- 8.8 years. Actual survival rate from diagnosis of bladder cancer were 90.8%, 68.6%, 53.3% and 30.3%, after 1, 3, 5 and 10 years, respectively. Ten cases were dead by bladder cancer, 21 cases by complicated cancer and 16 cases by another cause. CONCLUSION: The incidence of double cancer with bladder cancer were increased. Prostate cancer, colorectal cancer and breast cancer were gradually increased as complicated organs in Japan. The prognosis of double cancer patients with bladder cancer was poor than single bladder cancer patients.

Adult↗

The adsorptive properties of hydroxyapatite to albumin, dextran and lipids.

Hydroxyapatite is widely utilized as a component in dental hygiene agents. The ability to adsorb and remove dental plaque adhering to tooth surfaces is recognized as an important property of hydroxyapatite particles. In this study, the adsorptive ability of hydroxyapatite was evaluated based on its adsorptive amount and adsorptive strength. The adsorptive amount was determined by measuring the weight of albumin, dextran and lipids adsorbing to hydroxyapatite. The adsorptive strength was determined by measuring the weights of albumin and dextran remaining adsorbed after rinsing. Similar procedures were performed in control experiments using calcium hydrogen phosphate dihydrate, alumina and sericite. The results showed that the weights of albumin and dextran adsorbed to hydroxyapatite were 1.4 +/- 0.13 and 1.6 +/- 0.30 times higher than that to calcium hydrogen phosphate dihydrate. The weight of lipids adsorbed to hydroxyapatite was much higher than that to alumina and sericite. The percentages of weight loss of albumin and dextran were 2.3 +/- 1.7% and 5.3 +/- 2.0% for hydroxyapatite and 27 +/- 6.8% and 19 +/- 11% for calcium hydrogen phosphate dihydrate after one rinse. It is concluded that hydroxyapatite is a better adsorbent than calcium hydrogen phosphate dihydrate, alumina and sericite with respect to adsorbing albumin, dextran and lipids.

Adsorption↗

Minimum light intensity required to suppress nocturnal melatonin concentration in human saliva.

We set out to determine the minimum intensity of light able to suppress nocturnal melatonin levels as measured in normal human saliva. Five healthy male volunteers were exposed to light at different intensities (<10, 500, 1000, 2500, and 5000 lux) in a repeated measure design. Suppression of melatonin was dependent on both light intensity and duration of light exposure. Minimum intensities of light suppressing nocturnal melatonin levels were calculated as 393, 366, 339, and 285 lux for exposure durations of 30, 60, 90, and 120 min, respectively. Minimum effective intensity and duration of light exposure showed a linear inverse relationship. These results suggest that less intensity of light than previously reported suffices to suppress melatonin in humans, and that caution is required in interpreting studies using long exposure to dim light as a background condition.

Adult↗

Angiotensin II activates RhoA in cardiac myocytes: a critical role of RhoA in angiotensin II-induced premyofibril formation.

The organization of actin into striated fibers (myofibrils) is one of the major features of cardiac hypertrophy. However, its signal transduction mechanism is not well understood. Although Rho-family small G proteins have been implicated in actin organization in many cell types, it is not fully elucidated whether Rho mediates the organization of actin fibers by hypertrophic stimuli in cardiac myocytes. Therefore, we examined (1) whether Rho is activated by the hypertrophic stimulus, angiotensin II (Ang II), and (2) whether Rho mediates the Ang II-induced organization of actin fibers in cultured neonatal rat cardiac myocytes. Treatment of myocytes with Ang II caused a rapid formation of both striated (mature myofibrils) and nonstriated (premyofibrils) actin fibers within 30 minutes, as determined by phalloidin stainings of the polymerized actin and troponin T stainings. Immunoblot analyses and immunostainings have indicated that cardiac myocytes express RhoA, but RhoB is undetectable. In the control state, RhoA was observed predominantly in the cytosolic fraction, but it was translocated in part to the particulate fraction in response to Ang II, consistent with activation of RhoA by Ang II. Incubation of myocytes with exoenzyme C3 for 48 hours completely ADP-ribosylated Rho in vivo. The C3 treatment abolished formation of premyofibrils induced by Ang II, suggesting that Ang II causes premyofibril formation via a Rho-dependent mechanism. The Ang II-induced mature myofibril formation was only partly abolished by C3. Expression of constitutively active RhoA (V14RhoA) caused the formation of premyofibrils but not mature myofibrils. The C3 treatment inhibited Ang II-induced atrial natriuretic factor induction, whereas it had no effect on c-fos induction. These results indicate that RhoA is activated by Ang II and mediates the Ang II-induced formation of premyofibrils and induction of a subset of genes. Distinct signaling mechanisms seem to be responsible for striated mature myofibril formation by Ang II.

ADP Ribose Transferases↗

Localization of carbohydrate chains of pig sperm ligand in the glycoprotein ZPB of egg zona pellucida.

The three glycoproteins of pig zona pellucida (ZPA, ZPB and ZPC) can be separated into ZPA and a mixture of ZPB/ZPC by gel-filtration HPLC. We have shown previously that the neutral complex-type N-linked carbohydrate chains obtained from ZPB/ZPC possess sperm-binding activity. Intact ZPB and ZPC cannot be separated from each other unless acidic N-acetyllactosamine regions of their carbohydrate chains are removed by endo-beta-galactosidase digestion. The endo-beta-galactosidase-digested ZPB retains the sperm-binding activity. Recently, we have reported that N-linked carbohydrate chains of N-terminal fragment (residues 137-247) obtained from endo-beta-galactosidase-digested ZPB are involved mainly in sperm binding [Yonezawa, N., Mitsui, S., Kudo, K. & Nakano, M. (1997) Eur. J. Biochem. 248, 86-92]. In this study, we separated the intact neutral N-linked chains from the ZPB/ZPC mixture into diantennary chains and triantennary and tetraantennary chains by affinity chromatography on Concanavalia ensiformis agglutinin. An in vitro competition assay revealed that triantennary and tetraantennary chains possess a sperm-binding activity stronger than that of diantennary chains. Three glycopeptides, having one Asn residue to which the carbohydrate chain is linked, were obtained by lysyl endopeptidase digestion of the heat-solubilized zonae containing intact ZPB and lysyl endopeptidase and chymotrypsin A digestion of endo-beta-galactosidase-digested ZPB. From sugar-mapping analysis of the carbohydrate chains from these glycopeptides and comparison with the carbohydrate structures of the main intact neutral N-linked chains of ZPB/ZPC, the triantennary and tetraantennary chains were shown to be localized mainly at Asn220 of ZPB, and diantennary chains were present on all the three potential residues (Asn203, Asn220 and Asn333). These results suggest that the carbohydrate chains linked to Asn220 of ZPB participate predominantly in sperm-egg binding.

Amino Acid Sequence↗