Kinetics of phase separation and coarsening in dilute surfactant pentaethylene glycol monododecyl ether solutions.

We investigated the phase separation phenomena in dilute surfactant pentaethylene glycol monodedecyl ether (C(12)E(5)) solutions focusing on the growth law of separated domains. The solutions confined between two glass plates were found to exhibit the phase inversion, characteristic of the viscoelastic phase separation; the majority phase (water-rich phase) nucleated as droplets and the minority phase (micelle-rich phase) formed a network temporarily, then they collapsed into an usual sea-island pattern where minority phase formed islands. We found from the real-space microscopic imaging that the dynamic scaling hypothesis did not hold throughout the coarsening process. The power law growth of the domains with the exponent close to 1/3 was observed even though the coarsening was induced mainly by hydrodynamic flow, which was explained by Darcy's law of laminar flow.

Melting of polymer single crystals studied by dynamic Monte Carlo simulations.

We report dynamic Monte Carlo simulations of lattice polymers melting from a metastable chain-folded lamellar single crystal. The single crystal was raised and then melted in an ultrathin film of polymers wetting on a solid substrate, mimicking the melting observations made by using Atomic Force Microscopy. We observed that the thickness distribution of the single crystal appears quite inhomogeneous and the thickness increases gradually from facetted edges to the center. Therefore, at low melting temperatures, melting stops at a certain crystal thickness, and melting-recrystallization occurs when allowing crystal thickening; at intermediate temperatures, melting maintains the crystal shape and exhibits different speeds in two stages; at high temperatures, fast melting makes a melting hole in the thinnest region, as well as a saw-tooth-like pattern at the crystal edges. In addition, the linear melting rates at low temperatures align on the curve extrapolated from the linear crystal growth rates. The temperature dependence of the melting rates exhibits a regime transition similar to crystal growth. Such kinetic symmetry persists in the melting rates with variable frictional barriers for c -slip diffusion in the crystal as well as with variable chain lengths. Visual inspections revealed highly frequent reversals upon melting of single chains at the wedge-shaped lateral front of the lamellar crystal. We concluded that the melting kinetics is dominated by the reverse process of intramolecular secondary crystal nucleation of polymers.

Plasminogen activator inhibitor-1 aids nerve growth factor-induced differentiation and survival of pheochromocytoma cells by activating both the extracellular signal-regulated kinase and c-Jun pathways.

Astrocytes are thought to be critical to neurons' surviving damage caused by ischemic stroke or other injury. Plasminogen activator inhibitor-1 is one of the active soluble factors released by astrocytes and regulates plasminogen activator-plasmin proteolytic sequence in the CNS as a serpin. In this study, we show that plasminogen activator inhibitor-1 can promote neurite outgrowth and survival of rat pheochromocytoma cells in serum-deprived conditions, and that this neuroprotective activity is correlated with enhanced activation of both extracellular signal-regulated kinases following a direct phosphorylation of nerve growth factor receptor, Trk A, and of c-Jun. Our results suggest that plasminogen activator inhibitor-1 can act as a neurotrophic factor, protecting neurons from serum deprivation-induced neuron death not only by compensating for nerve growth factor functions, but also by activating the c-Jun/activating protein-1 pathway.

Cytochrome P-450 in liver microsomes of streptozotocin-induced diabetic rats: purification and characterization.

Two diabetes-inducible forms of cytochrome P-450, named P-450ST-1 and -ST-2, were purified from the liver microsomes of streptozotocin-diabetic male rats by sodium cholate solubilization, octylamino-Sepharose 4B chromatography and high-performance liquid chromatography with DEAE-5PW and hydroxyapatite columns. The purified P-450 forms gave a single band each on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular weight of 48,500 for P-450ST-1 or 48,000 for P-450ST-2. The CO-reduced spectral maxima of P-450ST-1 and -ST-2 were at 451 nm. The two cytochromes had the low-spin state of heme in the oxidized form. Both P-450ST-1 and -ST-2 catalyzed the metabolism of aniline, benzphetamine, p-nitroanisole, testosterone and aminopyrine. However, the catalytic activity of P-450ST-2 for these substrates was apparently higher than that of ST-1. Analyses of the NH2-terminal amino-acid sequence and Western immunoblot showed that P-450ST-1 and -ST-2 differed structurally from each other. The catalytic activities, molecular weights, NH2-terminal sequences and/or immunochemical properties of P-450ST-1 and -ST-2 did not agree with those of the other cytochrome P-450 forms purified from diabetic rats previously.

Puromycin aminonucleoside induces apoptosis and increases HNE in cultured glomerular epithelial cells(1).

Puromycin aminonucleoside induces apoptosis and increases 4-hydroxy-2-nonenal (HNE) in cultured glomerular epithelial cells. We have previously reported the detachment of cultured glomerular epithelial cells (GECs) from their substrata by puromycin aminonucleoside (PAN) treatment. In this study we explored whether or not apoptosis was involved in the mechanisms of the detachment. DNA fragmentation on gel electrophoresis was clearly shown by 10(-3) M PAN treatment of GECs. Nuclear staining by Hoechst 33342 indicated the greatest number of apoptotic cells at 10(-3) M PAN for 48 h treatment. Similarly, TUNEL methods revealed maximal apoptotic cells at 10(-3) M PAN for 48 h treatment. Caspase-3 (like) protease activity increased at 10(-3) M PAN, and decreased at 2 x 10(-3) M PAN for 48 h treatment as well as at 10(-3) M PAN for 60 h treatment. Pretreatment with 2'-deoxycoformycin (DCF), inhibitor of adenosine deaminase, abolished these effects of PAN on cultured GECs. PAN treatment increased HNE, a lipid peroxide adduct, modified protein in cultured GECs, which was also prevented by pretreatment by DCF. These results for the first time indicate that the PAN-induced detachment of GECs from culture substrata is mediated at least in part through apoptosis via oxidative stresses by adenosine deaminase activity.

Highly efficient oligosaccharide synthesis on water-soluble polymeric primers by recombinant glycosyltransferases immobilised on solid supports.

Recombinant beta-1,4-galactosyltranferase (beta 1,4-GalT) and alpha-2,6-sialytransferase (alpha 2,6-SiaT) immobilised covalently with activated Sepharose beads were employed for the practical synthesis of a trisaccharide derivative, Neu-5Ac alpha(2-->6)Gal beta(1-->4)GlcNAc beta-O-(CH2)6-NH2, on a water-soluble primer having GlcNAc residues through a alpha-chymotrypsin-sensitive linker.

Effect of bile acids on biliary excretion of cyclosporin A in the rat.

Cyclosporin A, a substrate of P-glycoprotein (P-gp), is known to cause cholestasis in humans and in rat experimental models. Tauroursodeoxycholate is reported to be effective in CyA-induced cholestasis in rats. In the present study, to investigate the mechanism of the inhibition of CyA induced cholestasis, effect of bile acids on biliary cyclosporin A excretion was studied in rats. Infusion of both taurocholate and tauroursodeoxycholate at the rate of 0.8 mmol/min per 100 g bodyweight increased bile flow and biliary cyclosporin A excretion, and the extent was more prominent with tauroursodeoxycholate. It was suggested that these findings were caused by the enhanced vesicular targeting of P-gp to the canalicular membrane by bile acids, thus increasing the numbers of P-gp in the canalicular membrane.

Inhibition by nimesulide of prostaglandin production in rat macrophages.

We have investigated the inhibitory action of nimesulide (4-nitro-2-phenoxymethanesulfonanilide) on release of prostaglandin E2 (PGE2) from rat peritoneal exudated macrophages (macrophages) and its mechanism of action. PGE2 release from macrophages stimulated with opsonized zymosan (OPZ) were increased in the 20 h after stimulation, whereas no significant increase was noted in PGE2 release from unstimulated macrophages. Nimesulide caused a weak inhibition of PGE2 release from macrophages at 15 min after OPZ stimulation as compared with indomethacin, but nimesulide caused approximately the same strong inhibition as indomethacin at 10 h after OPZ stimulation. Cellular cyclooxygenase (COX) activity in macrophage at 10 h after OPZ stimulation was increased approximately seven times the COX activity in macrophages before OPZ stimulation. Nimesulide caused approximately the same strong inhibition of cellular COX activity as indomethacin at 10 h after OPZ stimulation. COX-1 mRNA was expressed in macrophages irrespective of OPZ stimulation, but COX-2 mRNA was expressed only after OPZ stimulation, and COX-2 protein was simultaneously induced. Nimesulide affected neither the levels of COX-1 mRNA and COX-2 mRNA at 4 h after OPZ stimulation nor the levels of COX-2 protein at 10 h after OPZ stimulation. In contrast, actinomycin D caused strong inhibition of COX-2 mRNA expression and protein induction. These results suggest that inhibition by nimesulide of PGE2 release from macrophages, namely inflammatory cells, would be neither due to inhibition of COX-2 mRNA expression nor COX-2 induction, but to the selective inhibition of COX-2 activity itself.

Cross-sectional and longitudinal studies on the effect of water exercise in controlling bone loss in Japanese postmenopausal women.

The effects of water exercise, as a form of daily physical activity (water exercise in a warm water pool), on bone loss in healthy Japanese postmenopausal women have been cross-sectionally and longitudinally investigated from the viewpoint of preventing osteoporosis. In the cross-sectional study, the bone mineral density (BMD) of the lumbar spine (Z-score (%)) in the Veterans group (N = 27), who had been exercising for 35.2 months on average, was significantly higher than that in the Newcomers (N = 40), who had only begun to exercise 3 or 4 weeks before, and that in the Non-exercisers (N = 30), serving as a control group. The rate of change in the BMD of the lumbar spine was -0.92%/year in the Non-exercisers (N = 30), +1.55%/year in the Veterans (N = 20), and +2.16%/year in the Newcomers (N = 15), based on BMD Z-scores. In the Exercisers groups, it was found that the rate of change in the BMD showed a slight increase rather than a decrease irrespective of the duration of menopause. On the other hand, in the Non-exercisers group, the rate of change in the BMD decreased slightly. The results of questionnaires showed that the subjects' general awareness of health and fitness in daily life was enhanced after starting the water exercise program. These results suggest that consistently participating in water exercise is an important factor in preventing bone loss, and moreover, appears not only to indirectly improve awareness of daily physical activity but also to promote health and improve daily life.

[Early and late survival of total aortic arch replacement in patients with dissecting or atherosclerotic aneurysm].

Forty-seven patients (pts) were underwent total aortic arch replacement (TAAR) were studied to compare the early and late results between atherosclerotic (AA) and dissecting aneurysm (DA). There were 23 pts with AA and 24 pts with 24 pts. Preoperative risk were observed with shock in 12 pts (26%), major organ ischemia in 2 pts (4%), rupture in 8 pts (17%), stroke in 8 pts (17%) and coronary artery disease in 4 pts (9%). Operative procedure was TAAR in 22 pts, TAAR with ascending aorta replacement in 14 pts, and TAA with descending aorta replacement in 11 pts. Hospital mortality was 21 pts (45%) and late mortality was a surgical death after thoracoabdominal aneurysm in 1 pts. There was no difference in early and late survival rate, however early mortality was higher in pts with ruptured AA and with DA suffered from preoperative shock. There were 19 of early death in recent pts without rupture in AA and preoperative shock in DA.

[Successful emergent replacement of ruptured aortic arch aneurysm in an inpatient waiting for surgery: report of a case].

A 71-year-old male was admitted to our hospital for examination of mediastinal abnormal shadow, which was diagnosed aortic arch aneurysm. Once he was discharged and stayed at home for 2 weeks, and then re-admitted for surgery. On the 3rd day of 2nd admission, he suddenly fainted away in the ward and was in shock by hemorrhagic cardiac tamponade. He was transferred to operation room within an hour from onset of rupture. Emergent aortic arch replacement was performed under circulatory arrest with deep hypothermia and retrograde cerebral perfusion. His postoperative course was uneventful without any neurological deficits. Prompt diagnosis and surgery may contribute to improvement of surgical result in patients with ruptured aortic arch aneurysms.

High oleic/stearic fatty-acid desaturation index in cord plasma from infants of mothers with gestational diabetes.

OBJECTIVE: Enhanced fatty-acid desaturation by stearoyl-CoA desaturase enzyme-1 (SCD1) is associated with obesity. This study determined desaturation in the cord plasma of newborns of mothers with and without gestational diabetes (GDM). STUDY DESIGN: Newborns of mothers with GDM (n=21) and without (control, n=22) were recruited. Cord plasma fatty-acid desaturation indices (palmitoleic/palmitic, oleic/stearic ratios) were compared, and correlated with anthropometrics and biochemical measures. A subset of very low-density lipoprotein (VLDL) desaturation indices were determined to approximate the liver SCD1 activity. RESULT: The total oleic/stearic index was higher in GDM, despite adjustment for cord glucose concentrations. Among GDM and controls, the oleic/stearic index correlated with cord glucose concentrations (rs=0.36, P=0.02). Both palmitoleic/palmitic and oleic/stearic indices correlated with waist circumference (r=0.47, P=0.001; r=0.37, P=0.01). The VLDL oleic/stearic index was higher in GDM. CONCLUSION: The elevated total oleic/stearic index suggests increased lipogenesis in GDM newborns. Factors in addition to glucose supply may influence fetal SCD1 activity.

Plasma amino acid profile is associated with visceral fat accumulation in obese Japanese subjects.

UNLABELLED: What is already known about this subject • Asians with metabolic complications associated with obesity, a low body mass index and a low waist circumference have a greater proportion of visceral adipose tissue for a given amount of total body fat compared with Europeans. • Apparent obese humans and obese animal models show an elevation of branched-chain amino acid levels in plasma. • A multivariate logistic regression model of plasma free amino acids has been used to screen for several types of cancers in clinical settings. What this study adds • A specific formula incorporating six amino acid values (Ala, Gly, Glu, Trp, Tyr and branched-chain amino acid) was developed for discrimination of subjects with high visceral fat area by multivariate logistic regression analyses. • The generated amino acid formula was strongly correlated with visceral fat area in both apparent and non-apparent obese subjects. • Measuring plasma free amino acids can be used to distinguish the non-apparent visceral obesity in clinical settings in Asian populations. SUMMARY: Metabolic complications associated with obesity are becoming more common among Japanese subjects. However, visceral fat accumulation is not always apparent by measuring body mass index (BMI) or waist circumference in Asian populations because of the physiological characteristics particular to those ethnicities. Excess visceral fat accumulation raises the odds ratio for developing cardiovascular disease. Thus, high-throughput determination of the amount of abdominal adipose tissue is necessary. We hypothesized that accumulating visceral fat alters the peripheral amino acid profile and that a multivariate logistic regression model of plasma free amino acids can distinguish visceral obesity. A total of 1449 Japanese subjects (985 males and 464 females) who had undergone a comprehensive health screening were enrolled in this study. The visceral fat area was determined using computed tomography imaging, and a plasma free amino acid index to identify high visceral fat areas (≥100 cm(2) ) was developed. The sensitivity and specificity values of the generated amino acid index were 80% and 65%, respectively. In particular, the sensitivity of the generated index to identify subjects with non-apparent visceral obesity (BMI < 25 kg m(-2) ; visceral fat area ≥ 100 cm(2) ) was much greater than that of the waist circumference (73% vs. 46%, respectively). This index's high sensitivity and specificity may be the result of specific alterations in the patients' amino acid profiles, which were specifically correlated with the visceral fat areas and not with subcutaneous fat areas. This profile can be used as a predictor of elevated visceral obesity and a risk assessment tool for metabolic complications in Asian populations.

Antigenicity of sulfanilamide and its metabolites using fluorescent-labelled compounds.

In order to clarify the onset mechanisms of drug-induced allergies, three fluorescent-labelled compounds were synthesized by subjecting sulfanilamide (SA), a base compound for sulfonamides, and its active metabolites, i.e. sulfanilamide hydroxylamine and sulfanilamide nitroso, to dansylation using dansylchloride. In other words, 5-dimethylamino-N-(4-aminobenzyl)-naphthalenesulfonamide (DNS-4ABA), 5-dimethylamino-N-(4-hydroxylaminobenzyl)-1-naphthalenesulfonamide (DNS-4HABA) and 5-dimethylamino-N-(4-nitrosobenzyl)-1-naphthalenesulfonamide (DNS-4NSBA) were synthesized as model haptens. When analysed by HPLC, a conjugate of DNS-4HABA and glutathione (GSH) with nucleophilic amino acids had two peaks (P-1 and P-2). FAB-MS and 1H-NMR revealed that the DNS-4HABA-GSH conjugate consisted of sulphinamide and semimercaptal. The reactivity of GSH to DNS-4ABA, DNS-4HABA and DNS-4NSBA was quantified by HPLC using an oxidization system (horseradish peroxidase/H2O2). The results show that production of DNS-4NSBA-GSH-conjugate was four to eight times higher than that of DNS-4HABA-GSH conjugate, but that DNS-4ABA did not bind with GSH. Skin reactions were assessed using guinea pigs, and strong delayed erythema was seen with DNS-4NSBA, which bound most strongly with GSH, whereas weak delayed erythema was seen with DNS-4ABA, which did not bind with GSH. This suggests a correlation between GSH conjugate production and skin reactions. DNS-4HABA enzymatically bound with proteins in rat and guinea pig liver cytosol and microsomal fractions. The proteins that bound to DNS-4HABA were purified by HPLC and then subjected to N-terminal amino acid analysis. Ubiquitin (10 kDa) and fatty acid binding protein (30 kDa) were detected in the rat liver cytosol fraction; retinol-dehydrogenase (35 kDa) in the rat microsomal fraction; and glutathione-S-transferase B (mmu) (25 kDa) in the guinea pig liver cytosol fraction. When DNS-4HABA or DNS-4NSBA binds to proteins that play important roles in the body, unexpected adverse reactions may occur. Furthermore, by utilizing our technique using model compounds, it may be possible to identify the carrier proteins of various compounds, including pharmaceutical agents.

Nested polymerase chain reaction for detection of the avian leukosis virus causing so-called fowl glioma.

The complete nucleotide sequence of the avian leukosis virus causing so-called fowl glioma has been previously determined. Primers were designed for detection of the fowl glioma-causal virus (FGV) based on the 3' untranslated region of the viral genome. The provirus and viral RNA of FGV were specifically detected in various organs and tissues, including feather pulp, from experimentally infected birds using nested polymerase chain reaction (PCR) and reverse transcription nested PCR. The prevalence of FGV was evaluated in 131 Japanese fowls of a zoological garden in Japan based on the detection of the FGV genome in feather pulp using PCR and the detection of viral antigen in faeces by enzyme-linked immunosorbent assay. FGV proviral DNA was detected in feather pulp of 52 birds (39.7%) by nested PCR. Later, nine dead birds from among the 52 were histologically diagnosed as having fowl glioma and found to have the proviral DNA in the affected brain. These results demonstrated that the PCR-based detection of FGV in feather pulp is useful for epidemiological studies on fowl glioma.

Gene cluster for creatinine degradation in Arthrobacter sp. TE1826.

The genes encoding creatininase (CrnA; 258 residues) and creatinase (CreA; 411 residues) from Arthrobacter sp. TE1826 were cloned and sequenced. The genes form a cluster with the sarcosine oxidase gene (soxA) and its regulator gene (soxR), which were cloned previously. The deduced amino acid sequences of CrnA and CreA show 35.9% and 63.1% identity, respectively to the corresponding Pseudomonas enzymes. CrnA and CreA were purified from the recombinant strains and characterized. Other open reading frames (creB and crnB), encoding proteins similar to several transporters, were found downstream of creA and crnA, respectively.

Higher-order Laue zone line contrast in large-angle convergent-beam electron diffraction around a dislocation.

The physical picture of higher-order Laue zone (HOLZ) line contrast in a large-angle convergent-beam electron diffraction pattern around a dislocation, which is used for determining the Burgers vector, was examined. To evaluate the analytical expression of diffracted wave amplitude, we introduced an approximate form of the atomic displacement field of a dislocation. We showed that the four features of the HOLZ line contrast, that is, splitting, fading, bending and periodical contrast can be explained by analysis of the atomic displacement field. The localized lattice plane bending around a dislocation core made a HOLZ line split, fade and bend. However, we found that the periodical contrast of a HOLZ line was produced by the change of phase difference of the atomic displacement field between the crystals above and below the slip plane across the dislocation line.

Effects of experimental diabetes on aminopyrine metabolism in rats.

1. The metabolism of aminopyrine has been investigated in normal, alloxan- and streptozotocin (STZ)-diabetic rats. The drug was administered i.p. and the serum concentrations of the unchanged aminopyrine and its main metabolites were measured using h.p.l.c. 2. Aminopyrine was metabolized at a slower rate in both diabetic rats, as judged from higher serum levels of the unchanged drug. Pharmacokinetic studies of aminopyrine in diabetic rats also showed a decrease in serum clearance of the drug and an increase in its serum half-life. 3. The serum concentrations of the metabolites 4-monomethylaminoantipyrine, 4-acetylaminoantipyrine and 4-formylaminoantipyrine decreased in diabetic rats. In contrast, serum levels of 3-hydroxymethyl-2-methyl-4-dimethylamino-1-phenyl-3-pyrazolin-5-on e increased over control values. Serum concentrations of 4-aminoantipyrine remained unaltered by the induction of diabetes. 4. The magnitudes of changes in serum levels of these metabolites were larger in alloxan-diabetes than in STZ-diabetes. 5. Additional support for changed metabolism of aminopyrine was obtained from the investigation of microsomal preparations from diabetic and normal rats. 6. These findings indicate that it is important to use intact animals for evaluation of the metabolism of drugs in pathological states.