Inhibition of voltage-dependent calcium channels by prostaglandin E2 in rat melanotrophs

The effects of PGE2 on voltage-dependent Ca2+ channel currents were studied in dissociated rat melanotrophs by the whole-cell configuration of the patch-clamp technique. In about 90% of melanotrophs examined, PGE2 reversibly inhibited voltage-dependent Ba2+ currents elicited by voltage steps from a holding potential of -80 to 0 mV, with an ED50 of 68 nM. The maximum inhibition of Ba2+ currents by 1 microM PGE2 (35.3%) was comparable with that by the maximally effective concentration (100 nM) of dopamine. The EP1/EP3 PGE (EP) agonists, 17PT-PGE2 and sulprostone, and the EP2/EP3 agonist, misoprostol, mimicked the inhibition by PGE2, whereas the selective EP2 agonist, butaprostol, had little effect. The inhibition by PGE2 was partially, but significantly, reduced by the selective EP1 antagonist, SC-51322. The magnitude of the PGE2-induced inhibition of Ba2+ currents was greatly reduced by pretreatment with pertussis toxin, or by a depolarizing prepulse, to +80 mV, lasting for 50 msec. Although four distinct types (N-, P/Q-, L-, and R-types) of high-threshold Ba2+ currents were observed, PGE2 (1 microM) caused significant inhibition of only P/Q- and L-type currents, which were 17.3 and 10.1%, respectively, of the total Ba2+ currents. These results suggest that PGE2 inhibits P/Q- and L-type Ca2+ channels of rat melanotrophs via EP1 and EP3 receptors, which are coupled to pertussis toxin-sensitive G proteins, and produces both voltage-sensitive and -insensitive inhibition of Ca2+ channels.     

Influence of detergent builders on surface properties of sodium dodecyl sulfate solutions under dynamic and static conditions

The effects of various detergent builders on surface properties of sodium dodecyl sulfate (SDS) solutions have been investigated by measuring the equilibrium surface tension and hysteresis curves of surface pressure vs surface area under dynamic conditions. While the adsorbed layer of SDS under static conditions is in the liquid-expanded state, a two-dimensional phase transition of the adsorbed layer from liquid-expanded to condensed-film can be observed on compression at higher rates beyond the saturated adsorption. For the SDS solution containing excess NaCl (1×10⁻¹ M), the adsorbed layer of SDS is already in the condensed state under static condition due to a depression in the dissociation of SDS. It has been found that some polyelectrolytes, as well as conventional ionic builders, contribute to increasing the surface excess concentration of SDS together with promoting micelle formation and also to stabilizing the adsorbed layer of SDS. On the other hand, poly(vinyl alcohol) (PVA) as a nonionic polymer is adsorbed at the air-water interface in preference to SDS. In this case, the hysteresis of the dynamic surface behavior of the mixed solution can be attributed to the structural change of the adsorbed layer of PVA rather than SDS, which is different from the other ionic builders.     

POLYCYCLIC AROMATIC HYDROCARBON CONCENTRATIONS OF AIRBORNE PARTICLES IN URBAN AIR OVER THE PAST TWENTY YEARS

To estimate human exposure to hazardous chemicals, it is important to measure the annual trends regarding the concentrations of these compounds in the environment. Airborne particulate samples were collected over the past 20 years (1980–2001) from ambient air using a quartz fiber filter with the help of a high-volume air sampler placed on the top of the National Institute of the Public Health building, Minato-ku, Tokyo, Japan. In the present study, portions of these filter samples were analyzed to investigate the trends regarding the concentration of PAHs in urban air over the sampling period. The concentrations of various PAHs per unit air volume were determined. The concentrations of seven PAHs decreased within the sampling periods, especially in the late 1980s, and slowly decreased from the middle 1990s. The concentrations of seven PAHs were higher in autumn and winter than in the spring and summer.     

Quantitation of NAD+ biosynthesis from the salvage pathway in Saccharomyces cerevisiae

Nicotinamide adenine dinucleotide (NAD⁺) is synthesized via two major pathways in prokaryotic and eukaryotic systems: the de novo biosynthesis pathway from tryptophan precursors, or the salvage biosynthesis pathway from either extracellular nicotinic acid or various intracellular NAD⁺ decomposition products. NAD⁺ biosynthesis via the salvage pathway has been linked to an increase in yeast replicative lifespan under calorie restriction (CR). However, the relative contribution of each pathway to NAD⁺ biosynthesis under both normal and CR conditions is not known. Here, we have performed lifespan, NAD⁺ and NADH (the reduced form of NAD⁺) analyses on BY4742 wild‐type, NAD⁺ salvage pathway knockout (npt1Δ) and NAD⁺ de novo pathway knockout (qpt1Δ) yeast strains cultured in media containing either 2% glucose (normal growth) or 0.5% glucose (CR). We have utilized ¹⁴C labelled nicotinic acid in the culture media combined with HPLC speciation and both UV and ¹⁴C detection to quantitate the total amounts of NAD⁺ and NADH and the amounts derived from the salvage pathway. We observed that wild‐type and qpt1Δ yeast exclusively utilized extracellular nicotinic acid for NAD⁺ and NADH biosynthesis under both the 2% and 0.5% glucose growth conditions, suggesting that the de novo pathway plays little role if a functional salvage pathway is present. We also observed that NAD⁺ concentrations decreased in all three strains under CR. However, unlike the wild‐type strain, NADH concentrations did not decrease and NAD⁺: NADH ratios did not increase under CR for either knockout strain. Lifespan analyses revealed that CR resulted in a lifespan increase of approximately 25% for the wild‐type and qpt1Δ strains, while no increase in lifespan was observed for the npt1Δ strain. In combination, these data suggest that having a functional salvage pathway is required for lifespan extension under CR. Copyright © 2009 John Wiley & Sons, Ltd.     

Phase Transformations and Structural Developments in the Radular Teeth of Cryptochiton Stelleri

During mineralization, the hard outer magnetite‐containing shell of the radular teeth of Cryptochiton stelleri undergoes four distinct stages of structural and phase transformations: (i) the formation of a crystalline α‐chitin organic matrix that forms the structural framework of the non‐mineralized teeth, (ii) the templated synthesis of ferrihydrite crystal aggregates along these organic fibers, (iii) subsequent solid state phase transformation from ferrihydrite to magnetite, and (iv) progressive magnetite crystal growth to form continuous parallel rods within the mature teeth. The underlying α‐chitin organic matrix appears to influence magnetite crystal aggregate density and the diameter and curvature of the resulting rods, both of which likely play critical roles in determining the local mechanical properties of the mature radular teeth.     

Intercalation: Building a Natural Superlattice for Better Thermoelectric Performance in Layered Chalcogenides

A natural superlattice with composition (SnS)_1.2(TiS₂)₂, built by intercalating a SnS layer into the van der Waals gap of layered TiS₂, has been directly observed by high-resolution transmission electron microscopy (HRTEM). The thermoelectric performance is improved in the direction parallel to the layers because the electron mobility is maintained while simultaneously suppressing phonon transport, which is attributed to softening of the transverse sound velocities due to weakened interlayer bonding. In the direction perpendicular to the layers, the lattice thermal conductivity of (SnS)_1.2(TiS₂)₂ is even lower than the predicted minimum thermal conductivity, which may be caused by phonon localization due to the translational disorder of the SnS layers parallel to the layers. Moreover, we propose a large family of misfit-layer compounds (MX)_1+x (TX₂) _n (M = Pb, Bi, Sn, Sb, rare-earth elements; T = Ti, V, Cr, Nb, Ta; X = S, Se; n = 1, 2, 3) with a natural superlattice structure as possible candidate high-performance thermoelectric materials.     

Determination of enzymes from Colletotrichum sp. AHU9748 essential for lepidimoide production from okra polysaccharide

The allelopathic substance lepidimoide (Lp), which exhibits multiple functions in the growth and development of plants, was produced by Colletotrichum sp. AHU9748 from okra polysaccharide. Okra polysaccharide has the repeating structure (1-->4)-O-alpha-(d-galactopyranosyluronic acid)-(1-->2)-O-alpha-l-rhamnopyranose in its hexasaccharide repeating unit of its main chain. To determine the enzymes essential for Lp production, the supernatant of a culture broth was fractionated by repeated column chromatographies to identify two serial fractions responsible for Lp production and non-Lp production by measuring Lp production together with beta-galactosidase (beta-gal), rhamnogalacturonan lyase (RG-lyase) and acetylesterase (AE) activities, which we hypothesized to be necessary for Lp production from the structure of Lp. We confirmed the presence of these three enzymatic activities in the highest-Lp-producing fraction. The addition of purified RG-lyase to fractions producing no or a small amount of Lp demonstrated that beta-gal and RG-lyase activities are necessary for Lp production. The N-terminal amino acid sequences of the three separated proteins on SDS-PAGE confirmed the presence of enzymes identical to beta-gal, RG-lyase and AE in the Lp-producing fractions.     

Morphologic changes in boar sperm nuclei with reduced disulfide bonds in electrostimulated porcine oocytes

In pigs, failure of sperm nuclear decondensation has been reported after injection into oocytes. We examined the effects of pretreating sperm heads with Triton X-100 (TX-100) and dithiothreitol (DTT) and of electrical stimulation of oocytes after sperm head injection on time-dependent morphologic changes in sperm nuclei and in vitro development to the blastocyst stage. In experiment 1, spermatozoa were pretreated with 1% TX-100 and 5 mM DTT (T + D) or not treated, and then injected into in vitro matured oocytes. Electrical stimulation (1.5 kV/cm, 20 mus DC pulse) was applied to the oocytes 1 h after injection (stimulated group) or was not applied (unstimulated group). Some of the oocytes in each group were evaluated at hourly intervals until 10 h after injection for morphologic changes in the sperm nuclei. Unstimulated oocytes injected with untreated spermatozoa showed a delayed peak in the rate of nuclear decondensation (39.4-44.1%, 3-6 h after injection) compared with oocytes injected with T + D-treated spermatozoa (57.0% and 52.6%, 1 and 2 h, respectively). The rate of male pronucleus formation peaked 6 h after stimulation (by 40-60%) after injected oocytes had been stimulated with an electrical pulse, irrespective of whether or not the spermatozoa had been pretreated. In unstimulated oocytes, the rate of male pronucleus formation did not increase and stayed at the basal level (less than 20%) throughout the culture period, regardless of the sperm treatment. Thus, T + D treatment of spermatozoa did not affect completion of fertilization. In experiment 2, we evaluated the effects of electrical stimulation and sperm treatment with T + D on the rate of blastocyst formation and the mean number of cells per blastocyst. Oocytes stimulated after injection with either T + D-treated or untreated spermatozoa showed significantly higher percentages of blastocyst formation (24.8% and 27.1% respectively) than did unstimulated oocytes (1.1% and 4.1% for T + D-treated and untreated respectively; P < 0.01 by Duncan's multiple-range test). The rate of blastocyst formation did not differ between the T + D-treated and untreated groups. The mean number of cells per blastocyst did not differ among any of the groups (14.0-29.4 cells). These results suggest that pretreatment of sperm with TX-100 and DTT shifted the timing of sperm nuclear decondensation forward. However, pronucleus formation and development to the blastocyst stage in vitro were not improved by sperm treatment. Thus, electrical stimulation of injected oocytes enhances in vitro development to the blastocyst stage in pigs.     

Action of 1-(11-selenadodecyl)-glycerol and 1-(11-selenadodecyl)-3-trolox-glycerol against lipid peroxidation

The antioxidant action on lipid peroxidation of the synthesized selenium compounds 1-(11-selenadodecyl)-glycerol (SeG) and 1-(11-selenadodecyl)-3-Trolox-glycerol (SeIrG, where Trolox=6-hydroxyl-2,5,7,8-tetramethylchroman-2-carboxylic acid) was investigated. We compared the reactivity of the selenium compounds toward peroxyl radicals and their inhibitory effect on lipid peroxidation, induced by several kinds of initiating species such as azo compounds, metal ions, and superoxide/nitric oxide in solution, micelles, membranes, and rat plasma. SeTrG, but not SeG, scavenged peroxyl radicals. SeG reduced methyl linoleate hydroperoxides in organic solution and in methyl linoleate micelles oxidized by ferrous ion (Fe²⁺)/ascorbic acid. In rat plasma SeG and SeTrG decreased the formation of lipid hydroperoxides generated by hydrophilic azo compounds. SeG and SeTrG spared α-tocopherol (α-TOH) consumption in multilamellar vesicle membranes oxidized by hydrophilic or lipophilic initiators, and only SeTrG spared α-TOH in superoxide/nitric oxide oxidized membranes. In rat plasma oxidized by radical initiators (either hydrophilic or lipophilic) or superoxide/nitric oxide, SeTrG suppressed α-TOH consumption, but SeG had no effect. The two selenium-containing compounds showed inhibitory effects on lipid peroxidation that depended on their structure, the medium where they acted, and the oxidant used.     

Growth mechanism of AlGaAs on terraced substrates by low pressure MOVPE

AlGaAs layers were grown on recessed GaAs substrates by MOVPE at 5 and 100 Torr. The two mechanisms, the gas phase diffusion through the stagnant layer and the surface migration of the growing species, are responsible for the surface step height after the growth. Since the mean free path in the gas phase at 5 Torr (≈20μm) is longer than the recess height (≈1μm), only the surface migration determines the growth at 5 Torr, while both mechanisms contribute to the growth at 100 Torr. The surface diffusion equation is solved to find out the relation between the growth conditions and the surface step height after the growth. It was found that the surface migration length on the (111)A surface is much longer than that on the (100) plane. The optical waveguide is fabricated by growing a double-heterostructure on the recessed substrate, and light confinement in the channel is verified. On leave from Matsushita Kotobuki Electronics Ltd. Fukutake, Saijo 793, Japan