Biomedical subjects

T Inoue

Publications and source records attributed to T Inoue.

At least 631 records · Page 35Linked to original sources

PubMed1999

Evaluation of In-111 DTPA-paclitaxel scintigraphy to predict response on murine tumors to paclitaxel.

UNLABELLED: Our goal was to determine whether scintigraphy with 111In-DTPA-paclitaxel could predict the response to chemotherapy with paclitaxel. METHODS: Ovarian carcinoma (OCA 1), mammary carcinoma (MCA-4), fibrosarcoma (FSA) and squamous cell carcinoma (SCC VII) were inoculated into the thighs of female C3Hf/Kam mice. Mice bearing 8 mm tumors were treated with paclitaxel (40 mg/kg). The growth delay, which was defined as the time in days for tumors in the treated groups to grow from 8 to 12 mm in diameter minus the time in days for tumors in the untreated control group to reach the same size, was measured to determine the effect of paclitaxel on the tumors. Sequential scintigraphy in mice bearing 10 to 14 mm tumors was conducted at 5, 30, 60, 120, 240 min and 24 hrs postinjection of 111In-DTPA-paclitaxel (3.7MBq) or 111In-DTPA as a control tracer. The tumor uptakes (% injection dose/pixel) were determined. RESULTS: The growth delay of OCA 1, MCA-4, FSA and SCC VII tumors was 13.6, 4.0, -0.02 and -0.28 days, respectively. In other words, OCa 1 and MCA-4 were paclitaxel-sensitive tumors, whereas FSA and SCC VII were paclitaxel-resistant tumors. The tumor uptakes at 24 hrs postinjection of In-111 DTPA paclitaxel of OCA 1, MCA-4, FSA and SCC VII were 1.0 x 10(-3), 1.6 x 10(-3), 2.2 x 10(-3) and 9.0 x 10(-3) % injection dose/pixel, respectively. There was no correlation between the response to chemotherapy with paclitaxel and the tumor uptakes of 111In-DTPA-paclitaxel. CONCLUSIONS: Scintigraphy with 111In-DTPA-paclitaxel could not predict the response to paclitaxel chemotherapy. Although there was significant accumulation of the paclitaxel in the tumor cells, additional mechanisms must be operative for the agent to be effective against the neoplasm. 111In-DTPA-paclitaxel activity is apparently different from that of paclitaxel with Cremophor.

Animals↗

PubMed1999

Effect of edetate calcium disodium on yttrium-90 activity in bone of mice.

The kinetics of Yttrium-90 (Y-90) in bone of mice was investigated in combination with edetate calcium disodium (CaNa2EDTA). One group of mice were intraperitoneally administered 37.5 mg/kg CaNa2EDTA or 0.9% NaCl as a control at 1, 22, 34, 46, 58, 70, 82, 94, 154 and 166 h after injection of Y-90 acetate (post-administration), and the biodistribution was studied at 3, 24, 72, 120 and 168 h postinjection of Y-90 acetate. No difference between the post-CaNa2EDTA-treated mice and the control was demonstrated in the radioactivity in the bone. A decrease in radioactivity in the liver and kidneys was accelerated, and the radioactivity was lower than the control at 120 h postinjection. The other group of mice were also given the same dose of chelator at 12 h and 1 h preinjection of Y-90 acetate and at 1, 22, 34, 46, 58, 70, 82, 94, 154 and 166 h after injection of Y-90 acetate (pre- and post-administration), the radioactivity in bone at 3 h postinjection was significantly lower than in the control (24.4 +/- 3.92% ID/g vs. 31.7 +/- 2.26% ID/g, p < 0.05), but the decrease was not sequential. A significant reduction in radioactivity in the blood, kidneys and liver was demonstrated at 3 h, 72 h and 72 h postinjection. In conclusion, the CaNa2EDTA with the administration schedule employed here cannot chelate the Y-90 from bone but the free Y-90 before deposition into bone.

Animals↗

PubMed1999

Performance stability of SHR-2000 high resolution PET for animal research.

The performance of a high resolution positron emission tomography (PET) system SHR-2000 for animal studies was re-evaluated six years after its installation. The system employs a detector array consisting of BGO crystals that are 1.7 mm (transaxially) by 10 mm (axially) by 30 mm (deep). A block detector, which is a position-sensitive photomultiplier tube (PMT) coupled to 4 arrays of BGO crystals has been adopted to the system. There are 15 block detectors positioned to form a 35 cm diameter ring with a field of view (FOV) of 17 cm by 4.6 cm axially, giving the system a 7 slice imaging capability. For six year workload in spatial resolution (FWHM), there were approximately a 2.6% increase at tangential FOV and a 7.5% increase at radial FOV. In axial resolution (FWHM) there was almost no change. The count rate loss for the true count rate increased 1.3% at 200 kBq/ ml. The average slice sensitivity showed a decrease of approximately 4.1%, and in scatters it showed an increase of approximately 1.4%. In animal experiments, the bones of guinea pigs were clearly identified with 18F fluoride ion. These experiments show that after a six year workload, the system also maintains good performance and has good stability.

Animals↗

PubMed1999

Spectroscopic and electrochemical properties of two azurins (Az-iso1 and Az-iso2) from the obligate methylotroph Methylomonas sp. strain J and the structure of novel Az-iso2.

Two azurin-type blue copper proteins, which are related to the electron-transfer processes involving methylamine/methanol oxidation, have been spectroscopically and electrochemically characterized. The obligate methylotroph Methylomonas sp. strain J gives rise to two azurins (Az-isol and Az-iso2) with methylamine dehydrogenase (MADH-Mj). The intense blue bands characteristic of Az-iso1 and Az-iso2 are observed at 621 and 616 nm in the visible absorption spectra respectively, being revealed at 620-630 nm in those of usual azurins. The EPR signal of Az-iso1, similar to usual azurins, shows axial symmetry, while the axial EPR signal of Az-iso2 involves a slightly rhombic character. The half-wave potentials (E1/2) of the two azurins and the intermolecular electron-transfer rate constants (kET) from MADH-Mj to each azurin were determined by cyclic voltammetry. The E1/2 values of Az-iso1 and Az-iso2 are +321 and +278 mV vs NHE at pH 7.0, respectively. The kET value of Az-iso2 is larger than that of Az-iso1 by a factor of 5. However, the electron-transfer rate of Az-iso2 is interestingly slower than those of the azurins from a denitrifying bacterium, Alcaligenes xylosoxidans NCIB 11015, and the amicyanin from a different methylotroph, Methylobacterium extorquens AM1. The structure of Az-iso2 has been determined and refined against 1.6 A X-ray diffraction data. The whole structure of Az-iso2 is quite similar to those of azurins reported already. The Cu(II) site of Az-iso2 is a distorted trigonal bipyramidal geometry like those of other azurins, but some of the Cu-ligand distances and ligand-Cu-ligand bond angle parameters are slightly different. These findings suggest that Az-iso2 is a novel azurin and perhaps functions as an electron acceptor for MADH.

Azurin↗

PubMed1999

Effect of an intermittent weekly dose of human parathyroid hormone (1-34) on osteoporosis: a randomized double-masked prospective study using three dose levels.

To test the effect of amino-terminal peptide 1-34 of human parathyroid hormone (hPTH (1-34)) as a possible bone anabolic agent in the treatment of osteoporosis, weekly subcutaneous injection of 50 units (L group), 100 units (M group) or 200 units (H group) of hPTH (1-34) was started in 220 patients with osteoporosis at 71 institutions randomly divided into three groups in a double-masked system. Lumbar spine bone mineral density (BMD) by dual-energy X-ray absorptiometry (DXA) increased by 0.6%, 3.6% and 8.1% after 48 weeks in groups L, M and H respectively, responses in groups M and H being significantly higher than in L (p<0.05, Mann-Whitney U-test). Since the coefficient of variation for lumbar spine measurement stayed at 1-2.5%, increases of 3.6% and 8.1% appeared significant. Metacarpal BMD and cortical thickness measured by radiogrammetry did not change significantly. Serum calcium decreased in each group and serum phosphorus decreased in groups M and H. Urinary calcium/creatinine decreased at the 12th week in group H and at the 24th and 48th weeks in groups M and L. Serum 25(OH) vitamin D and 1,25(OH)(2) vitamin D decreased in each group at the 48th week (p<0.05). Serum bone-type alkaline phosphatase was increased at the fourth week in groups H and M and decreased at the 48th week in group H. Urinary hydroxyproline, pyridinoline and deoxypyridinoline declined significantly in each group. Backache improved in 30-40% of each group. No serious adverse effects were found during the test period. Intermittent weekly injection of hPTH (1-34) increased lumbar BMD in osteoporosis, suggesting usefulness in the treatment of osteoporosis.

Aged↗

PubMed1999

The relationships between the degree of beta-isomerization of type I collagen degradation products in the urine and aging, menopause and osteoporosis with fractures.

We have evaluated the effect of aging, menopause and osteoporosis on the measurements of both nonisomerized type I collagen C-telopeptide breakdown products (alpha-CTx) by radioimmunoassay (RIA) and beta-isomerized type I collagen C-telopeptide breakdown products (beta-CTx) by enzyme-linked immunosorbent assay (ELISA). In 86 premenopausal healthy women (PRE), 144 postmenopausal healthy women (POST), 74 patients with vertebral fractures (VX) and 61 patients with hip fractures (HX), urinary CTx excretion was measured by both ELISA and RIA assays. Samples were collected more than 6 months after fracture in the VX group and within 48 h after fracture in the HX group. In all subjects a highly significant correlation was found between alpha-CTx and beta-CTx (r = 0.85). The values of beta-CTx in the POST group greatly increased compared with those in the PRE group (% mean increase: 82%), while the values of alpha-CTx in the POST group moderately increased compared with those in the PRE group (% mean increase: 47%). The values of both alpha-CTx and beta-CTx in the HX group were significantly higher than those in the other groups, but particularly the increase in mean alpha-CTx (211% for HX versus POST) was very high compared with the increase in mean beta-CTx (68% for HX versus POST). Moreover, the alpha-CTx/beta-CTx ratio in the HX group was significantly higher than in the other groups. These results suggest that both assays well reflect the increase in bone resorption associated with high bone turnover, especially, in osteoporotic patients with hip fracture. However, there was a difference between the urinary excretion of alpha-CTx and beta-CTx in patients with hip fracture, so the alpha-CTx/beta-CTx radio might be a good indicator reflecting the characteristics of bone metabolism for osteoporosis with hip fracture.

Adolescent↗

PubMed1999

A double-masked multicenter comparative study between alendronate and alfacalcidol in Japanese patients with osteoporosis. The Alendronate Phase III Osteoporosis Treatment Research Group.

To evaluate the efficacy and safety of alendronate, a double-masked, active (alfacalcidol) controlled comparative study for 48 weeks was carried out in a total of 210 Japanese patients with osteoporosis. The doses of alendronate and alfacalcidol were 5 mg/day and 1 microgram/day, respectively. The lumbar bone mineral density (LBMD) values observed at 12, 24, 36 and 48 weeks after the initiation of alendronate treatment were 3.53 +/- 0.53%, 5.37 +/- 0.62%, 5.87 +/- 0.74% and 6.21 +/- 0.59% (mean +/- SE), respectively, higher than the baseline value. Corresponding values in the alfacalcidol group were 1.50 +/- 0.43%, 0.69 +/- 0.63%, 1.12 +/- 0.60% and 1.36 +/- 0. 63%, respectively. There was a significant difference between the two groups at each time point (p<0.05 or p<0.001). The bone turnover markers were depressed during treatment in the alendronate group: -32.2% for alkaline phosphatase, -53.7% for N-terminal osteocalcin and -45.0% for urinary deoxypyridinoline compared with the corresponding baseline values. On the contrary, no notable changes in these parameters were observed in the alfacalcidol group. Treatment with alendronate caused a transient decrease in serum calcium concentrations associated with an increase in the serum level of intact parathyroid hormone. In contrast, treatment with alfacalcidol resulted in a tendency of these parameters to change in the opposite direction. No difference in fracture incidence between the two groups was observed. The overall safety of alendronate was comparable to that of alfacalcidol. In conclusion, although it was a relatively short-term study of 48 weeks, the results of the present study indicate that alendronate at the daily dose of 5 mg was effective in increasing LBMD and that no serious drug-related adverse events were observed in the alendronate-treated patients. Alendronate is more efficacious than alfacalcidol in increasing bone mineral density, although the mechanisms of the actions of the two drugs are apparently different.

Aged↗

PubMed1999

Calcaneus bone mineral density using single x-ray absorptiometry in Japanese women.

Calcaneus bone mineral density (BMD) of 738 Japanese women (605 healthy and 133 with osteoporosis) was measured using single X-ray absorptiometry (SXA). A reference range of calcaneus BMD values for healthy Japanese women was established and the usefulness of this method for screening and diagnosis of osteoporosis was evaluated. There was no significant age change of calcaneus BMD prior to menopause, though values decreased significantly thereafter. BMD loss ratio was 1.7%/year in the 10 years after menopause. The reference range of calcaneus BMD was 410 +/- 43 mg/cm(2), calculated from the mean BMD value of subjects whose ages ranged from 25 to 50 years old. The fracture threshold for the spine was established as 294 mg/cm(2), which corresponded to -2.67 SD from the average BMD of the young healthy women, and the odds ratio for spine fracture in the subjects with BMD lower than this threshold was 3.52 [95% CI (confidence interval) 1.34-9.26]. The spine fracture group showed statistically lower calcaneus BMD than the nonfracture group when subjects with adjusted age and body size were analyzed. There were no significant differences in the ROC analysis for spine fracture between calcaneus BMD and spine BMD. Therefore, calcaneus BMD is not readily affected by degenerative change or soft tissue, and the annual decrement rate (1.7%/year) can be detected easily and with low precision error (0.8%). These indices may prove useful for the screening and diagnosis of osteoporosis.

Absorptiometry, Photon↗

PubMed1999

An alternative approach to estimation of the brain perfusion index for measurement of cerebral blood flow using technetium-99m compounds.

Cerebral blood flow (CBF) has been quantified non-invasively using the brain perfusion index (BPI) determined from radionuclide angiographic data generated by technetium-99m hexamethylpropylene amine oxime( )((99m)Tc-HMPAO) or technetium-99m ethyl cysteinate dimer( )((99m)Tc-ECD). The BPI is generally calculated using graphical analysis (GA). In the present study, BPI was measured using spectral analysis (SA), and its usefulness evaluated in comparison with GA. The BPI was calculated from the sum of spectral data obtained by SA. We applied this method to radionuclide angiographic data collected from the bilateral brain hemispheres of 20 patients with various brain diseases using (99m)Tc-HMPAO and from those of 20 patients using (99m)Tc-ECD. We also measured BPI using GA. The BPI values obtained by SA (BPI(S)) (x) and by GA (BPI(G)) (y) correlated closely (y=0.708x+0.038, r=0.945 for (99m)Tc-HMPAO and y=0.559x+0.093, r=0.931 for (99m)Tc-ECD). However, the BPI(G) values were underestimated by 22.9%+/-6.6% (mean+/-SD) for (99m)Tc-HMPAO and by 27.9%+/-7.5% for (99m)Tc-ECD as compared with the BPI(S) values. The extent of underestimation tended to increase with increasing BPI(S) values. These findings were considered to be a result of the BPI(G) values being affected by the first-pass extraction fraction of the tracer. We also compared the BPI(S) and BPI(G) values with those of CBF measured using N-isopropyl-p-[(123)I]iodoamphetamine (CBF(IMP)) in 16 patients (six for (99m)Tc-HMPAO and ten for (99m)Tc-ECD). Although both BPI(S) and BPI(G) values correlated significantly with the CBF(IMP) values, the correlation coefficient in BPI(S) was always better than that in BPI(G) (r=0.869 for (99m)Tc-HMPAO and r=0.929 for (99m)Tc-ECD in BPI(S), r=0.629 for (99m)Tc-HMPAO and r=0.856 for (99m)Tc-ECD in BPI(G)). These results suggest that SA can provide a more reliable BPI for quantifying CBF using (99m)Tc-HMPAO or (99m)Tc-ECD than the conventional method using GA. Our method will be useful especially when using a tracer with a low first-pass extraction fraction and/or when performing activation studies using pharmacological intervention.

Aged↗

PubMed1999

Effects of glucocorticoids on bone mineral density in rheumatoid arthritis patients. A longitudinal study.

We carried out a comparative study in 78 post-menopausal women with rheumatoid arthritis (RA). Forty-four women with a mean disease duration of 17.5 years had been treated with low-dose glucocorticoid (prednisone at < 5 mg/day) for at least 12 months. They were studied for an average period of 3 years and 8 months. The remaining 34 women had been treated only with nonsteroidal anti-rheumatic drugs (NSAIDs) and served as the control group. Bone mineral density (BMD) in the lumbar spine (L2-4) and femoral neck was measured by dual-energy X-ray absorptiometry (DXA). Reduction of BMD in the lumbar spine was significant in both groups (P < 0.05 to approximately 0.01), but there was no statistically significant difference between the two groups. BMD of the femoral neck decreased significantly (P < 0.05) in the prednisone group, but again the difference was not significant between the two groups. Our data suggest that low-dose prednisone administration probably does not induce significant axial bone loss in female RA patients.

Absorptiometry, Photon↗

PubMed1999

Changes of regional distribution of bone mineral according to age, gender, and activity.

To assess the pattern of bone mineral distribution over various parts of the skeletal system depending on age, gender, and level of activity, bone mineral content of the head, arms, trunk, and legs was measured and expressed as percentage of the whole-body mineral content (H%, A%, T%, and L%) by a QDR-4,500 (Hologic) in 814 subjects between ages of 20 and 99, 205 men and 609 women. In ambulant, self-supporting subjects classified as ADL 1, women had higher H%, lower A%, and lower L% at each age group than in men. In subjects older than 60 years of age, H% increased and L% decreased markedly in women, whereas these changes were milder in men. Women, but not men, showed definite fall of L% and rise of H% along with the decrease of the level of activity. In hospitalized female subjects with a lower level of activity, but not in ambulatory female subjects, fall of L% and rise of H% was noted after 1 year, unlike the results in their male counterparts. Mineral distribution changes over the whole body characterized by age, gender, and activity may serve as new indices for evaluation of skeletal changes caused by aging, lifestyle, and disease.

Activities of Daily Living↗

PubMed1999

Position-independent human beta-globin gene expression mediated by a recombinant adeno-associated virus vector carrying the chicken beta-globin insulator.

The position-independent expression of transgenes in target cells is an essential subject for determining effective gene therapies. The chicken beta-globin insulator blocks the effects of regulatory sequences on transcriptional units at differential domains. We prepared a recombinant adeno-associated virus (rAAV) containing various combinations of the DNase I-hypersensitive site 2 (HS2), 3 (HS3), and 4 (HS4) core elements from the human beta-globin locus control region (LCR), the human beta-globin gene, and the herpes virus thymidine kinase promoter driven neomycin-resistant gene (neoR) (rHS432, rHS43, rHS42, rHS32, and rHS2), and also rAAV containing two copies of the 250-bp core sequence of the chicken beta-globin insulator on both sides of the rHS2 (rIns/HS2/2Ins). After isolating neomycin-resistant mouse erythroleukemia (MEL) cells infected with each rAAV, we analyzed the rAAV genome by Southern blots and polymerase chain reaction (PCR), using primers specific for HS core elements and the human beta-globin gene. All clones contained a single copy of the rAAV genome in the chromosome, however, some of them had a rearranged proviral genome. In five clones with a single unrearranged rAAV genome for each rAAV construct, we assayed the expression of the human b-globin gene relative to the endogenous mouse beta maj-globin gene, using quantitative reverse transcriptase (RT)-PCR. In clones infected with rHS432, the expression level of the human beta-globin gene ranged from 51.6% to 765.6% of that in the mouse beta maj-globin gene. Likewise, in rHS43, the expression level ranged from 36.7% to 259.0%; in rHS42, from 47.8% to 207.0%; in rHS32, from 47.9% to 105.4%; and in rHS2, from 6.1% to 172.1%, indicating a high variability of expression level in clones infected with recombinant virus lacking the insulator. In contrast, in clones infected with rIns/HS2/Ins, the range of expression of the human beta-globin gene ranged from 52.8% to 58.3% of that in the mouse beta maj-globin gene. These results indicate that the insulator functioned dramatically to reduce the variability of transgene expression due to the position effect. This insulator-rAAV vector system holds promise to provide a constant level of transgene expression for gene therapy, regardless of the insertion sites on the chromosome.

Animals↗

PubMed1999

Does pressure antagonize anesthesia? High-pressure stopped-flow study of firefly luciferase and anatomy of initial flash.

The antagonizing effect of high pressure against anesthesia is well known. With purified firefly luciferase, however,. Biophys. J. 60:1309-1314) reported that high pressure did not affect the initial flash intensity. Firefly luciferase emits a burst of light when the substrates luciferin and ATP are added in the presence of O2. The light intensity decays rapidly and the weak light lasts for hours. The initial flash is a transient event and is not in a steady state. The steady state is represented by the slope of the linear part of the integral of the light output. The present study used a high-pressure stopped-flow system to compare the pressure effects on the initial flash intensity and the steady-state light intensity. The flash intensity did not change by the application of hydrostatic pressure in the presence or absence of chloroform or 1-octanol. In contrast, high pressure increased the steady-state light intensity. The application of 12 MPa pressure increased the steady-state light intensity of firefly luciferase inhibited by 5 mM chloroform or 0.7 mM 1-octanol by 19.7% and 18.8%, respectively. When analyzed by the rapid reaction kinetics of the transition state theory, the initial peak intensity represents the total amount of active enzyme and is unrelated to the reaction rate. Anesthetics inhibited the initial flash by unfolding the protein, thereby decreasing the concentration of the active enzyme. Pressure affected the steady-state light intensity by changing the reaction rates.

Anesthesia↗

PubMed1999

Morphological behavior of acidic and neutral liposomes induced by basic amphiphilic alpha-helical peptides with systematically varied hydrophobic-hydrophilic balance.

Lipid-peptide interaction has been investigated using cationic amphiphilic alpha-helical peptides and systematically varying their hydrophobic-hydrophilic balance (HHB). The influence of the peptides on neutral and acidic liposomes was examined by 1) Trp fluorescence quenched by brominated phospholipid, 2) membrane-clearing ability, 3) size determination of liposomes by dynamic light scattering, 4) morphological observation by electron microscopy, and 5) ability to form planar lipid bilayers from channels. The peptides examined consist of hydrophobic Leu and hydrophilic Lys residues with ratios 13:5, 11:7, 9:9, 7:11, and 5:13 (abbreviated as Hels 13-5, 11-7, 9-9, 7-11, and 5-13, respectively; Kiyota, T., S. Lee, and G. Sugihara. 1996. Biochemistry. 35:13196-13204). The most hydrophobic peptide (Hel 13-5) induced a twisted ribbon-like fibril structure for egg PC liposomes. In a 3/1 (egg PC/egg PG) lipid mixture, Hel 13-5 addition caused fusion of the liposomes. Hel 13-5 formed ion channels in neutral lipid bilayer (egg PE/egg PC = 7/3) at low peptide concentrations, but not in an acidic bilayer (egg PE/brain PS = 7/3). The peptides with hydrophobicity less than Hel 13-5 (Hels 11-7 and Hel 9-9) were able to partially immerse their hydrophobic part of the amphiphilic helix in lipid bilayers and fragment liposome to small bicelles or micelles, and then the bicelles aggregated to form a larger assembly. Peptides Hel 11-7 and Hel 9-9 each formed strong ion channels. Peptides (Hel 7-11 and Hel 5-13) with a more hydrophilic HHB interacted with an acidic lipid bilayer by charge interaction, in which the former immerses the hydrophobic part in lipid bilayer, and the latter did not immerse, and formed large assemblies by aggregation of original liposomes. The present study clearly showed that hydrophobic-hydrophilic balance of a peptide is a crucial factor in understanding lipid-peptide interactions.

Biophysical Phenomena↗

PubMed1999

Tolerability, pharmacokinetics, and pharmacodynamics of DX-9065a, a new synthetic potent anticoagulant and specific factor Xa inhibitor, in healthy male volunteers.

OBJECTIVE: The aim of this study was to assess the tolerability, pharmacokinetic and pharmacodynamic properties of DX-9065a, a novel low-molecule specific factor Xa inhibitor in healthy male volunteers. METHODS: DX-9065a was intravenously administered to healthy male Japanese volunteers at doses of 0.625 to 30 mg. The drug concentrations in plasma and urine were measured and pharmacokinetic parameters were calculated. Coagulation time and bleeding time were also measured. RESULTS: No serious adverse event was observed during or after administration of DX-9065a. The pharmacokinetics of DX-9065a in human subjects after intravenous dosing was linear. The simulated plasma concentrations of DX-9065 were well in accordance with the observed values. Though prolongation of coagulation times was dependent on plasma concentration of DX-9065, bleeding time did not increase even at the highest plasma concentration of 1640 ng/mL. CONCLUSIONS: DX-9065a had a good correlation between linear pharmacokinetics and pharmacodynamics after intravenous administration in humans.

Adult↗

PubMed1999

Recombinant adenovirus vectors for cytokine gene therapy in mice.

BACKGROUND: Adenoviruses have several specific features useful for gene therapy. They infect various lineages of cells irrespective of cell cycle status. However, the exact mechanism of their infection and in vivo kinetics as a gene expression vector have not been elucidated. OBJECTIVE: Using adenovirus vectors expressing marker genes, we examined the infectivity of these vectors (including cellular and tissue tropism), the duration and intensity of transgene expression, and the side effects. METHODS: Various cells were infected with adenovirus expressing LacZ gene at various doses, and beta-galactosidase activity was measured and compared in relation with dose, time course, and cellular vitronectin receptor. Mice were injected with adenoviruses expressing LacZ, luciferase and GM-CSF, and in vivo gene expression was examined. RESULTS: Adenovirus infection induced viral dose-dependent transgene expression that persisted for 2 weeks. Adherent cells were infected much more efficiently than nonadherent cells, probably because the former expressed much higher levels of the vitronectin receptor, one of the main receptors for adenovirus. Studies performed in mice with luciferase-expressing adenovirus revealed that the liver was the main target organ after intravenous injection and showed that the intravenous route was superior to other routes with regard to transgene expression. After intravenous injection of adenovirus expressing human GM-CSF, there was a transient and dose-dependent increase in the serum level of this cytokine. Administration of adenovirus expressing mouse GM-CSF enhanced hematopoiesis in the spleen and bone marrow. CONCLUSION: These results indicated that adenoviruses can be used for in vivo cytokine gene therapy but suggested the necessity of taking into consideration the route of administration, the duration of transgene expression, the toxic dose, and host immune reactions.

Adenoviridae↗

PubMed1999

Nonpeptide mimic of bradykinin with long-acting properties.

Kinins, members of a family of peptides released from kininogens by the action of kallikreins, have been implicated in a variety of biological activities including vasodilation, increased vascular permeability, contraction of smooth muscle cells and activation of sensory neurons. However, investigation of the physiological actions of kinins have been greatly hampered because its effects are curtailed by rapid proteolytic degradation. We examined the pharmacological characteristics of the first nonpeptide bradykinin receptor agonist 8-[2,6-dichloro-3-[N-[(E)-4-(N-methylcarbamoyl)cinnamidoacetyl+ ++]-N-methylamino]benzyloxy]-2-methyl-4-(2-pyridylmethoxy)quinolin e (FR190997). FR190997, whose structure is quite different from the natural peptide ligand, but is similar to the nonpeptide antagonists FR165649, FR167344 and FR173657, potently and selectively interacts with the human B2 receptor and markedly stimulates inositol phosphate formation in transfected Chinese hamster ovary (CHO) cells. FR190997 induces concentration-dependent contraction of isolated guinea pig ileum. In vivo, FR190997 mimics the biological action of bradykinin and induces hypotensive responses in rats with prolonged duration, presumably as a consequence of its resistance to proteolytic degradation. Therefore, FR190997 is a highly potent and subtype-selective nonpeptide agonist which displays high intrinsic activity at the bradykinin B2 receptor. This compound represents a powerful tool for further investigation of the physiology and pathophysiology of bradykinin receptors.

Animals↗

PubMed1999

An enhancement of nitric oxide production regulates energy metabolism in rat hepatocytes after a partial hepatectomy.

BACKGROUND/AIMS: Infection after a liver resection often results in hepatic failure. Nitric oxide is one of the candidates which has been suspected to cause cellular dysfunction during infection in the liver. We have previously reported that the inflammatory cytokine interleukin-1beta (IL-1beta) induced the expression of the inducible nitric oxide synthase gene in primary cultured rat hepatocytes. We hypothesized that an enhancement of nitric oxide production after the resection was implicated in a change in liver energy metabolism, thus resulting in liver dysfunction. METHODS: In this study, we performed a 70% hepatectomy or a sham operation in rats, and then isolated hepatocytes from the remnant liver by collagenase perfusion. The cultured hepatocytes were treated with cytokines including IL-1beta. The effects on nitric oxide induction, the ATP content and ketone body ratio (acetoacetate/beta-hydroxybutyrate) were then compared between the partial hepatectomized (PH) and sham-operated (control) rats. RESULTS: IL-1beta augmented the induction of nitric oxide production two-fold in hepatocytes from the PH rats as compared to the control rats. IL-1beta markedly decreased the ATP content in the PH rats, although IL-1beta also decreased the ATP content in the control rats, but to a lesser extent. IL-1beta also decreased the ketone body ratio in both groups. The addition of L-arginine further stimulated the inhibition of the ATP levels and the ketone body ratio concomitantly with increased nitric oxide production in the PH rats. N(G)-monomethyl-L-arginine, an inhibitor of nitric oxide synthase, abolished the effects of IL-1beta on the ATP levels and ketone body ratio, as well as on the nitric oxide production. CONCLUSIONS: These results demonstrate that the decreased ATP content observed in PH rats resulted from an increase in nitric oxide production. The decrease in ketone body ratio indicates that nitric oxide-induced mitochondrial dysfunction contributes significantly to ATP attenuation in hepatocytes. Therefore, the regulation of nitric oxide induction may be crucial for preventing liver failure after a hepatic resection.

Adenosine Diphosphate↗