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T Kanaya

Publications and source records attributed to T Kanaya.

At least 19 recordsLinked to original sources
PubMed2005

Dynamics of deuterated polystyrene-protonated butadiene diblock copolymer micelles by neutron spin echo.

We report on neutron spin-echo (NSE) measurements on deuterated styrene-protonated butadiene diblock copolymer micelles in deuterated n-decane to investigate the dynamics of butadiene blocks in the corona. Before the NSE measurements, we performed small-angle neutron-scattering (SANS) measurements on the micelles to evaluate the structure to give a basis for the discussion of the dynamics. In the SANS study, we have estimated the form factor P(Q) in terms of a hard-core-shell model from the direct evaluation without curve-fitting procedure while a more flexible core-shell model with the structure factor S(Q) gives a better fit to the observed data. The observed normalized intermediate scattering function I(Q,t)/I(Q,0) by NSE does not show the collective motions corresponding to the so-called breathing mode but rather single chain motion (Zimm modes) for both the 2 and 20 wt % micelle solutions. The Zimm decay rate Gamma(z) in the micelle solution is slow compared with that in the homopolymer solution. This slowing down is assigned to the effective high concentration in the corona. The differences in Gamma(z) between concentrated solutions and the 20% micellar solution are attributed to end-tethering effect of the corona chains on the core surface. The possible reasons why the breathing mode was not observed in the present micelle system are discussed on the basis of chain density in the corona.

Journal Article↗
PubMed2005

Neutron spin-echo studies on dynamic and static fluctuations in two types of poly(vinyl alcohol) gels.

We report neutron spin-echo measurements on two types of poly(vinyl alcohol) (PVA) gels. The first is PVA gel in a mixture of dimethyl sulfoxide (DMSO) and water with volume ratio 60/40 , and the second is PVA gel in an aqueous borax solution. The observed normalized intermediate scattering functions I (Q,t) /I (Q,0) are very different between them. The former I (Q,t) /I (Q,0) shows a nondecaying component in addition to a fast decay, but the latter does not have the nondecaying one. This clearly indicates that the fluctuations in the former PVA gel consist of static and dynamic fluctuations whereas the latter PVA gel does include only the dynamic fluctuations. The dynamic fluctuations of the former and latter gels have been analyzed in terms of a restricted motion in the network and Zimm motion, respectively, and the origins of these motions will be discussed.

Journal Article↗
PubMed2005

Prevention of diet-induced obesity by dietary isomerized hop extract containing isohumulones, in rodents.

OBJECTIVE: Isomerized hop extract (IHE), which consists mainly of isohumulones and is required in the beer brewing process, was investigated for its effects on diet-induced obesity in two strains of mice. DESIGN: C57BL/6N and KK-A(y) mice were fed a standard or high-fat diet containing IHE and their body and tissue weights were measured at various time points. Oral glucose tolerance tests (OGTT) and insulin tolerance tests (ITT) were carried out in high-fat diet-fed C57BL/6N mice. The effects of IHE on intestinal lipid absorption were examined in Wistar rats using a plasma triacylglycerol assay after oral administration of a lipid emulsion. Fecal lipid levels were also measured in these animals after they were fed a high-fat diet containing IHE for 15 days. The effects of IHE on pancreatic lipase activity and the expression of genes involved in hepatic lipid metabolism were also examined using an in vitro assay and quantitative RT-PCR, respectively. RESULTS: Supplementation of high-fat-containing chow with IHE reduced body weight gain and improved glucose tolerance in our experimental mice. A reduction in body weight gain was also observed in C57BL/6N mice fed a standard diet containing IHE. Wistar rats fed a high-fat diet containing IHE showed reduced plasma triacylglycerol levels and an increase in their fecal lipid excretion. Similarly, their pancreatic lipase activity was inhibited and their elevation in plasma triacylglycerol levels seen after the oral administration of lipid emulsion was significantly suppressed. IHE-fed mice showed an increased expression in their lipid oxidation genes and a decreased expression in genes involved in triacylglycerol biosynthesis. CONCLUSION: The inhibition of intestinal dietary fat absorption may be the mechanism by which IHE induces its weight-lowering effects in high-fat diet-fed mice. The modulatory effect of IHE on lipid metabolism may also, at least partly, be responsible for its beneficial effects on body weight gain. These results suggest that IHE may be helpful in humans in preventing diet-induced obesity and perhaps even metabolic syndrome, the latter of which is known to be associated with obesity.

Animals↗
PubMed2004

Contraction and reexpansion of polymer thin films.

We report x-ray reflectivity measurements on polystyrene thin films supported on silicon wafer. In annealing experiments, we found fast and slow contraction processes in the thin films above the glass transition temperature. The former is the normal relaxation (annealing) process observed in bulk, and the latter is unexpected and enhanced in thin films below approximately 20 nm. In addition, we found unexpected extremely slow reexpansion processes in the glassy state. These unexpected very slow processes are discussed in terms of lateral contraction and expansion processes driven by entropic changes at the interfaces and the difference of the expansivities between polystyrene and silicon wafer.

Journal Article↗
PubMed2001

Apparent nonergodic behavior of supercooled liquids above the glass transition temperature.

A speckle pattern can be observed in the polarized component of light scattered from glass forming liquids far above their glass transition temperature. This speckle pattern fluctuates with characteristic time that corresponds to the relaxation time of the additional ultraslow component in the correlation function and is about seven orders of magnitude longer than the relaxation time of the alpha-process. This slow process is out of the experimental time window when the alpha-process is measured by means of the photon correlation spectroscopy and results in an apparent nonergodicity which can be seen as a baseline offset in the ensemble-averaged correlation function. In contrast, the time-averaged field correlation functions which have been measured in practically all light scattering studies always decay to zero. The slow process contributes a q-dependent excess intensity to the polarized component of scattered light. The values of the nonergodicity parameters obtained from the static and dynamic light scattering experiments are equal. Both the slow component and the excess intensity result from denser regions of fractal character which develop in glass-forming liquids on approaching the glass transition.

Journal Article↗
PubMed2001

Serological diagnosis of equine influenza using the hemagglutinin protein produced in a baculovirus expression system.

The hemagglutinin (HA) protein of an equine influenza strain, A/equine/La Plata/1/93 (LP/93), was produced using a baculovirus expression system. Silkworm larvae inoculated with recombinant baculovirus expressed high quantities of the HA protein which was then purified to greater than 95% purity by fetuin-affinity chromatography. Purified HA protein was used subsequently in an ELISA for detection of antibodies in horse sera. Two hundred serum samples from vaccinated racehorses were reacted on ELISA plates coated with 40.0 ng/ml of purified HA protein. Subsequent optical density (OD) levels revealed titers which correlated highly with respective hemagglutinin inhibition (HI) antibody titers which ranged from <1:8 to 1:256 (correlation coefficient among them was 0.850). ELISA OD levels and HI titers increased at 5 and 7 days post-inoculation, respectively, in a horse inoculated intranasally with LP/93. Respective antibody levels were observed to change in an essentially parallel manner during a period of 1 month. Similarly, ELISA OD levels correlated with HI titers in horses during a period of 6 weeks following intramuscular inoculation with inactivated single-strain vaccines containing LP/93, A/equine/Kentucky/1/81 (H3N8) or A/equine/Rome/5/91 (H3N8). A similar pattern was also observed in eight horses throughout a 10-week period following inoculation with a commercially available inactivated trivalent vaccine containing A/equine/Newmarket/1/77(H7N7), A/equine/Kentucky/81 and LP/93. From these results, it is suggested that this ELISA system could be used for disease diagnosis and surveillance of HI antibody titers among vaccinated horses.

Animals↗
PubMed2000

Identification and characterization of negative regulatory elements of the human telomerase catalytic subunit (hTERT) gene promoter: possible role of MZF-2 in transcriptional repression of hTERT.

Human telomerase reverse transcriptase (hTERT) is a catalytic subunit of human telomerase and is a critical determinant of the enzymatic activity of telomerase. Expression of hTERT is known to be regulated mainly at the transcriptional level. In the present study, using transient expression assays, we identified a 400 bp silencer of the hTERT promoter between -776 and -378 upstream of the proximal core promoter. The inhibitory effects of this silencer were enhanced with cellular differentiation. A computer-assisted homology search identified multiple binding motifs for myeloid-specific zinc finger protein 2 (MZF-2) within this region. Mutation introduced in these sites resulted in significant activation of hTERT transcription. Gel shift assays demonstrated that MZF-2 proteins specifically bound to these sites. Overexpression of MZF-2 in cells led to down-regulation of hTERT transcription as well as telomerase activity. These findings suggest that the 400 bp region upstream of the hTERT core promoter that we identified functions as a negative regulatory region and that MZF-2 may be an effector of negative regulation of hTERT.

Base Sequence↗
PubMed2000

Sp1 cooperates with c-Myc to activate transcription of the human telomerase reverse transcriptase gene (hTERT).

Telomerase activation is thought to be a critical step in cellular immortalization and carcinogenesis. The human telomerase catalytic subunit (hTERT) is a rate limiting determinant of the enzymatic activity of human telomerase. In the previous study, we identified the proximal 181 bp core promoter responsible for transcriptional activity of the hTERT gene. To identify the regulatory factors of transcription, transient expression assays were performed using hTERT promoter reporter plasmids. Serial deletion assays of the core promoter revealed that the 5'-region containing the E-box, which binds Myc/Max, as well as the 3'-region containing the GC-box, which binds Sp1, are essential for transactivation. The mutations introduced in the E-box or GC-box significantly decreased transcriptional activity of the promoter. Overexpression of Myc/Max or Sp1 led to significant activation of transcription in a cell type-specific manner, while Mad/Max introduction repressed it. However, the effects of Myc/Max on transactivation were marginal when Sp1 sites were mutated. Western blot analysis using various cell lines revealed a positive correlation between c-Myc and Sp1 expression and transcriptional activity of hTERT. Using fibroblast lineages in different stages of transformation, we found that c-Myc and Sp1 were induced to a dramatic extent when cells overcame replicative senescence and obtained immortal characteristics, in association with telomerase activation. These findings suggest that c-Myc and Sp1 cooperatively function as the major determinants of hTERT expression, and that the switching functions of Myc/Max and Mad/Max might also play roles in telomerase regulation.

Base Sequence↗
PubMed2000

A case of cervical cancer with aggressive tumor growth: possible autocrine growth stimulation by G-CSF and Il-6.

BACKGROUND: Patients with tumors that produce hematopoietic growth factors, such as G-CSF, sometimes exhibit leukemoid reactions. Squamous cell carcinomas of the uterine cervix producing such factors are rarely reported. CASE: A 56-year-old woman had severe leukocytosis (>50 x 10(3)/microl) and fever without evidence of infection after radical hysterectomy for treatment of her cervical cancer. The parametrial recurrent mass appeared after the operation and exhibited extremely aggressive growth. Serum levels of G-CSF and IL-6 were extremely high, and immunohistochemical analysis and RT-PCR assays revealed that G-CSF and IL-6 as well as their receptors were produced in tumor cells. CONCLUSIONS: This is a case of cervical cancer which exhibited an aggressive clinical course, possibly due to autocrine stimulation of cell growth by G-CSF and IL-6.

Carcinoma, Squamous Cell↗
PubMed2000

Acylated delphinidin 3-rutinoside-5-glucosides in the flowers of Petunia reitzii.

Two acylated anthocyanins were isolated from selected individuals of Petunia reitzii, and identified to be delphinidin 3-O-[6-O-(4-O-(4-O-(6-O-(trans-caffeoyl)-beta-D-glucopyranosyl)-tr ans-p-coumaroyl)-alpha-L-rhamnopyranosyl)-beta-D-glucopyranoside]- 5-O-[beta-D-glucopyranoside] and delphinidin 3-O-[6-O-(4-O-(4-O-(beta-D-glucopyranosyl)-trans-p-coumaroyl)-alph a-L-rhamnopyranosyl)-beta-D-glucopyranoside]-5-O-[beta-D-glucopyranoside ]. Nine known anthocyanins were also identified.

Acylation↗
PubMed2000

Purification and characterization of an insect haemolymph protein promoting in vitro replication of the Bombyx mori nucleopolyhedrovirus.

We have identified a novel protein that promotes Bombyx mori nucleopolyhedrovirus (BmNPV) replication in vitro. This protein was purified from heat-treated haemolymph of B. mori larvae by gel filtration and ion exchange chromatography, and designated as promoting protein (PP). The molecular mass of native PP estimated by column chromatography and that of denatured PP estimated by SDS-PAGE were 9600 Da and 15200 Da, respectively, suggesting that native PP is composed of a single polypeptide and may behave in the column as if it is a smaller protein because of its conformation and/or adsorptive nature. Addition of the PP to the culture medium of SES-BoMo-15A cells derived from B. mori embryos resulted in the strong promotion of BmNPV replication. The promoting activity positively correlated with the amount of PP in the culture medium up to 1 microg/ml, above which maximum virus replication occurred and resulted in the highest budded virus production and polyhedrin promoter-mediated luciferase gene expression of 10000-fold and 6000-fold higher than those without PP, respectively. A cDNA encoding the PP precursor (prePP) was successfully cloned and sequenced. Comparison between the amino acid sequence deduced from the nucleotide sequence of prePP cDNA and the N-terminal 18 amino acids determined for the purified PP indicated that the prePP (154 amino acids) consisted of a mature PP polypeptide (136 amino acids) with a signal sequence at the N terminus. Recombinant PP expressed from the cDNA using a baculovirus vector was similar in molecular mass, immunoreactivity and promoting activity to the native PP.

Amino Acid Sequence↗
PubMed2000

Ultra-small-angle neutron scattering studies on phase separation of poly(vinyl alcohol) gels

Time-resolved light scattering measurements during the gelation process of a poly(vinyl alcohol) (PVA) solution in a mixture of dimethyl sulfoxide and water (H2O) have shown that a spinodal decomposition (SD) type phase separation takes place in the early stage of gelation. In this case the kinetics of SD is valid only before macroscopic gelation occurs because the growth rate is slowed down by gelation. Such SD type phase separation makes the solution opaque as it proceeds, and hence the structural change can no longer be followed by light scattering. To investigate the structure of the opaque PVA gel as well, we have employed an ultra-small-angle neutron scattering technique using a Bonse-Hart camera. These observations reveal that even after the macroscopic gelation the structure due to the microphase separation on a spatial scale of several &mgr;m continues to grow against the elasticity. This may be because at first the gel structure is too soft to suppress the growth of the microphase separation, but within 24 h after the quenching the growth terminates. On the basis of the results, we will discuss a possible mechanism of the microphase separation after gelation.

Journal Article↗
PubMed2000

Further evidence of spinodal decomposition during the induction period of polymer crystallization: time-resolved small-angle x-ray scattering prior to crystallization of poly(ethylene naphthalate)

Aiming to clarify spinodal decomposition of polymers in the induction period of crystallization, time-resolved small-angle x-ray scattering measurements have been made in situ for poly(ethylene naphthalate) while it was crystallized from the glass, or in the case of the so-called glass crystallization. It is confirmed for this polymer that in the very beginning of the induction period a scattering peak appears at around 0.03 A (-1) in scattering vector q, which corresponds to a characteristic wavelength of 200 A in density fluctuations, and grows with time. Time evolution of this scattering peak is described by the kinetics of the spinodal decomposition as previously reported for the glass crystallization of poly(ethylene terephthalate).

Journal Article↗
PubMed2000

Adenoviral expression of p53 represses telomerase activity through down-regulation of human telomerase reverse transcriptase transcription.

Telomerase activation is a critical step in cellular immortality and oncogenesis. The activity of telomerase is known to be correlated with cell proliferation, but its regulation by cell cycle regulators is not well understood. In the present study, we examined the effects of p53 on telomerase activity. Wild-type p53 was introduced into SiHa cells via a recombinant adenoviral vector, Ad5CMV-p53, and change in telomerase activity was examined by quantitative telomerase assay. Telomerase activity in the Ad5CMV-p53-infected cells was significantly repressed 36 h after infection following down-regulation of human telomerase catalytic subunit [human telomerase reverse transcriptase (hTERT)] mRNA expression, whereas no change in telomerase activity was observed in the cells infected with control vector AdSCMV-beta-gal. Interestingly, repression of telomerase activity was an early event that preceded cell growth inhibition or apoptosis induced by p53 overexpression, suggesting that p53 directly regulates telomerase activity. Transient expression assays using hTERT-promoter reporter constructs revealed that overexpression of p53 significantly repressed promoter activity of hTERT. 5'-Truncation of the promoter sequences revealed that the proximal core promoter region containing multiple binding sites for transcription factor Spl was responsible for p53-mediated transcriptional repression. Mutations in these binding sites for Spl led to failure of p53 to repress transcription. These findings suggest that p53 repressed telomerase activity through down-regulation of hTERT transcription and that interaction of p53 with Sp1 or other transcription factors may be involved in this regulation.

Adenoviridae↗
PubMed2000

Demethylating reagent 5-azacytidine inhibits telomerase activity in human prostate cancer cells through transcriptional repression of hTERT.

Telomerase activation is thought to be a critical step in cellular immortality and oncogenesis. Several reagents including differentiation-inducing and antineoplastic agents are known to inhibit telomerase activity, although the molecular mechanisms through which they inhibit telomerase activity remain unclear. Demethylating reagents have recently been used as potential antineoplastic drugs for some types of cancers including those of the prostate. In the present study, we examined the effect of the demethylating reagent 5-azacytidine (5-aza-CR) on telomerase activity using cells of two prostate cancer cell lines, DU-145 and TSU-PR1. 5-aza-CR treatment significantly reduced telomerase activity in TSU-PR1 cells, but not in DU-145 cells, although growth inhibition was observed to a similar extent in both cell lines. Reverse transcription-PCR analyses revealed that inhibition of telomerase activity was accompanied by down-regulation of telomerase catalytic subunit (hTERT) mRNA expression. Transient expression assays showed that 5-aza-CR repressed the transcriptional activity of the hTERT promoter and that the E-box within the core promoter was responsible for this down-regulation. Western blot analyses revealed that 5-aza-CR reactivated p16 expression and repressed c-Myc expression in TSU-PR1 cells but not in DU-145 cells. Overexpression of p16 in TSU-PR1 cells led to significant repression of c-Myc transcription. These findings suggest that 5-aza-CR inhibits telomerase activity via transcriptional repression of hTERT, in which p16 and c-Myc may play a key role.

Azacitidine↗
PubMed1999

Estrogen activates telomerase.

Telomerase activity is present in most malignant tumors and provides a mechanism for the unlimited potential for division of neoplastic cells. Although telomerase is known to be a regulated enzyme, the factors and mechanisms involved in telomerase regulation are not well understood. In the present study, we examined the effects of estrogen on telomerase activity. Telomerase activity in estrogen receptor (ER)-positive MCF-7 cells was up-regulated by the treatment with 17beta-estradiol. This activation accompanied up-regulation of the telomerase catalytic subunit, hTERT mRNA. Gel shift assays revealed that the imperfect palindromic estrogen-responsive element in the hTERT promoter specifically binds to ER. Transient expression assays using luciferase reporter plasmids containing various fragments of hTERT promoter showed that this imperfect palindromic estrogen-responsive element is responsible for transcriptional activation by ligand-activated ER. We also found that estrogen activates c-Myc expression in MCF-7 cells and that E-boxes in the hTERT promoter that bind c-Myc/Max play additional roles in estrogen-induced transactivation of hTERT. Estrogen thus activates telomerase via direct and indirect effects on the hTERT promoter. These findings may help elucidate the mechanisms of hormonal control of telomerase activity and aid understanding of the roles of sex steroids in cellular senescence and aging as well as estrogen-induced carcinogenesis.

Base Sequence↗
PubMed1999

Preparation of recombinant rat interleukin-5 by baculovirus expression system and analysis of its biological activities.

Rat interleukin-5 (IL-5) cDNA was subcloned from peritoneal cells collected 4 h after intraperitoneal injection of Ascaris suum antigen solution into the immunized rats. Cysteine proteinase-deleted (CPd) rat IL-5 recombinant virus was constructed by inserting rat IL-5 cDNA into CPd virus having a deletion in the cysteine proteinase gene of the silkworm Bombyx mori nuclear polyhedrosis virus. On infection with the CPd rat IL-5 recombinant virus, the silkworm B. mori larvae produced rat IL-5 as a dimeric form in hemolymph. Recombinant rat IL-5 was purified more than 95.5% by anion-exchange chromatography and hydrophobic chromatography. The purified recombinant rat IL-5 promoted the proliferation of T88-M cells in a concentration-dependent manner, and its effect was inhibited by an anti-murine IL-5 neutralizing polyclonal antibody. When bone marrow cells from normal rats were incubated with recombinant rat IL-5 in medium containing methylcellulose, the colony formation by eosinophilic cells was induced. Furthermore, when rat peritoneal eosinophils were incubated with recombinant rat IL-5, the spontaneous decrease in the eosinophil viability was inhibited in time- and concentration-dependent manners. In addition, the recombinant rat IL-5-induced eosinophil survival was inhibited by an anti-murine IL-5 neutralizing polyclonal antibody. These findings suggest that rat IL-5 acts as B-cell growth factor II (BCGF-II), eosinophil differentiation factor (EDF), and eosinophil survival-enhancing factor.

Animals↗