Dopamine D1 receptor subtype mediates acute stress-induced dendritic growth in excitatory neurons of the medial prefrontal cortex and contributes to suppression of stress susceptibility in mice.

Dopamine in prefrontal cortices is implicated in cognitive and emotional functions, and the dysfunction of prefrontal dopamine has been associated with cognitive and emotional deficits in mental illnesses. These findings have led to clinical trials of dopamine-targeting drugs and brain imaging of dopamine receptors in patients with mental illnesses. Rodent studies have suggested that dopaminergic pathway projecting to the medial prefrontal cortex (mPFC) suppresses stress susceptibility. Although various types of mPFC neurons express several dopamine receptor subtypes, previous studies neither isolated a role of dopamine receptor subtype nor identified the site of its action in mPFC. Using social defeat stress (SDS) in mice, here we identified a role of dopamine D1 receptor subtype in mPFC excitatory neurons in suppressing stress susceptibility. Repeated social defeat stress (R-SDS) reduces the expression of D1 receptor subtype in mPFC of mice susceptible to R-SDS. Knockdown of D1 receptor subtype in whole neuronal populations or excitatory neurons in mPFC facilitates the induction of social avoidance by SDS. Single social defeat stress (S-SDS) induces D1 receptor-mediated extracellular signal-regulated kinase phosphorylation and c-Fos expression in mPFC neurons. Whereas R-SDS reduces dendritic lengths of mPFC layer II/III pyramidal neurons, S-SDS increases arborization and spines of apical dendrites of these neurons in a D1 receptor-dependent manner. Collectively, our findings show that D1 receptor subtype and related signaling in mPFC excitatory neurons mediate acute stress-induced dendritic growth of these neurons and contribute to suppression of stress susceptibility. Therefore, we propose that D1 receptor-mediated dendritic growth in mPFC excitatory neurons suppresses stress susceptibility.

Anomalous pressure effect in heteroacene organic field-effect transistors.

Anomalous pressure dependent conductivity is revealed for heteroacene organic field-effect transistors of dinaphtho[2, 3-b:2', 3'-f]thieno[3, 2-b]thiophene single crystals in the direction of a and b crystallographic axes. In contrast to the normal characteristics of a monotonic increase in mobility µ with the application of external hydrostatic pressure P in conductors, we found that the present organic semiconductor devices exhibit nonmonotonic and gigantic pressure dependence including an even negative pressure coefficient dµ/dP. In combination with a structural analysis based on x-ray diffraction experiments under pressure, it is suggested that on-site molecular orientation and displacement peculiar in heteroacene molecules are responsible for the anomalous pressure effect.

Light compensation points in shade-grown seedlings of deciduous broadleaf tree species with different successional traits raised under elevated CO2.

We measured leaf photosynthetic traits in shade-grown seedlings of four tree species native to northern Japan, raised under an elevated CO2 condition, to investigate the effects of elevated CO2 on shade tolerance of deciduous broadleaf tree species with different successional traits. We considered Betula platyphylla var. japonica and Betula maximowicziana as pioneer species, Quercus mongolica var. crispula as a mid-successional species, and Acer mono as a climax species. The plants were grown under shade conditions (10% of full sunlight) in a CO2 -regulated phytotron. Light compensation points (LCPs) decreased in all tree species when grown under elevated CO2 (720 µmol·mol(-1) ), which were accompanied by higher apparent quantum yields but no photosynthetic down-regulation. LCPs in Q. mongolica and A. mono grown under elevated CO2 were lower than those in the two pioneer birch species. The LCP in Q. mongolica seedlings was not different from that of A. mono in each CO2 treatment. However, lower dark respiration rates were observed in A. mono than in Q. mongolica, suggesting higher shade tolerance in A. mono as a climax species in relation to carbon loss at night. Thus, elevated CO2 may have enhanced shade tolerance by lowering LCPs in all species, but the ranking of shade tolerance related to successional traits did not change among species under elevated CO2 , i.e. the highest shade tolerance was observed in the climax species (A. mono), followed by a gap-dependent species (Q. mongolica), while lower shade tolerance was observed in the pioneer species (B. platyphylla and B. maximowicziana).

Photosynthetic traits of Siebold's beech seedlings in changing light conditions by removal of shading trees under elevated CO2.

The purpose of this study was to obtain basic information on acclimation capacity of photosynthesis in Siebold's beech seedlings to increasing light intensity under future elevated CO2 conditions. We monitored leaf photosynthetic traits of these seedlings in changing light conditions (before removal of shade trees, the year after removal of shade trees and after acclimation to open conditions) in a 10-year free air CO2 enrichment experiment in northern Japan. Elevated CO2 did not affect photosynthetic traits such as leaf mass per area, nitrogen content and biochemical photosynthetic capacity of chloroplasts (i.e. maximum rate of carboxylation and maximum rate of electron transport) before removal of the shade trees and after acclimation to open conditions; in fact, a higher net photosynthetic rate was maintained under elevated CO2 . However, in the year after removal of the shade trees, there was no increase in photosynthesis rate under elevated CO2 conditions. This was not due to photoinhibition. In ambient CO2 conditions, leaf mass per area and nitrogen content were higher in the year after removal of shade trees than before, whereas there was no increase under elevated CO2 conditions. These results indicate that elevated CO2 delays the acclimation of photosynthetic traits of Siebold's beech seedlings to increasing light intensity.

Separation and properties of the NAD-linked and NADP-linked isozymes of succinic semialdehyde dehydrogenase in Euglena gracilis z.

Euglena gracilis z contained two succinic semialdehyde dehydrogenases (EC 1.2.1.16), one requiring NAD and the other NADP, and these isozymes were separated from each other and partially purified. The NAD-linked isozyme was relatively stable on storage at 5 degrees C whereas the NADP-linked one was extremely unstable unless 30% glycerol or ethyleneglycol was added. The optimum pH was 8.7 and optimum temperature 35-45 degrees C for both isozymes. They were inhibited by Zn2+ and activated, particularly the NAD-linked enzyme, by K+. Sulfhydryl reagents activated both isozymes. The Km values for succinic semialdehyde were 1.66 - 10(-4) M with the NAD-linked isozyme and 1.06 - 10(-3) M with the NADP-linked one. The NADP-linked isozyme was induced by glutamate while the NAD-linked one was not. Probable roles of these isozymes in the physiology of Euglena gracilis are discussed.

Thiamin uptake in Euglena gracilis.

Thiamin uptake has been investigated in Euglena gracilis Z. This protozoon possessed an active transport system for thiamin with a Km value of 17 nM and a Vmax value of 7.8 pmol per 10(6) cells per min. Thiamin uptake was dependent on pH and temperature, but not on exogenous glucose as an energy source. Oxythiamin and pyrithiamin were competitive inhibitors with Ki values of 33 nM and 15 nM, respectively. Thiamin monophosphate, thiamin pyrophosphate, thiamin triphosphate, heteropyrithiamin, quinolinothiamin, thiamin chloride and amprolium inhibited uptake. Inhibition of thiamin uptake by various metabolic inhibitors and anaerobiosis suggest that thiamin uptake requires an energy source generated by respiration and glycolysis.

Effects of changes in the aortic input impedance on systolic pressure-ejected volume relationships in the isolated supported canine left ventricle.

We examined the effects of aortic input impedance alteration on left ventricular pressure, aortic flow and ejected volume (integral value of aortic flow), in an isolated blood perfused ejecting canine heart, with special reference to end-systolic values. A hydraulic model which stimulates an aortic input impedance was attached to the aortic root of an excised heart. Left ventricular end-diastolic pressure was kept constant by electrical pacing. Three coronary arteries were perfused with arterial blood from support dogs. When the peripheral resistance in the hydraulic model was changed, there were inverse linear relationships between stroke volume and mean left ventricular systolic pressure and between ejected volume and pressure at end-systole. Time interval from the onset of contraction to end-systole did not change. Thus the relation between stroke volume and mean left ventricular pressure obtained by changes in peripheral resistance is governed by a source resistance, which can be considered as the contractile state of the ventricle. When the capacitance (arterial compliance) was changed, there was no inverse linear relation between stroke volume and mean systolic pressure. In many cases, there was an inverse linear relationship between ejected volume and pressure at end-systole. However, an increase in capacitance prolonged the time interval from the onset of contraction to end-systole. We conclude that the end-systolic pressure-ejected volume relationship in the ejecting heart is governed not only by contractility but also by arterial capacitance.

Comparative efficacy of broad-spectrum antiviral agents as inhibitors of rotavirus replication in vitro.

Several nucleoside analogues which have previously been established as broad-spectrum antiviral agents, i.e. ribavirin, vidarabine, pyrazofurin, tubercidin, carbodine, (S)-9-(2,3-dihydroxypropyl)adenine [(S)-DHPA], carbocyclic 3-deazaadenosine (C-c3 Ado), (RS)-3-adenine-9-yl-2-hydroxypropanoic acid [(RS)-AHPA] isobutyl ester and neplanocin A were compared for their potency and selectivity as inhibitors of human rotavirus (strains Wa, KUN and MO) replication in vitro. As the most efficacious inhibitors emerged (S)-DHPA, C-c3 Ado, (RS)-AHPA isobutyl ester and neplanocin A, with a minimum inhibitory concentration of 60, 1.4, 1.2 and 0.2 micrograms/ml, and a selectivity index of greater than 3, 70, 80 and greater than 20, respectively. As has been postulated for their antiviral action in general, these adenosine analogues probably owe their anti-rotavirus activity to inhibition of S-adenosylhomocysteine hydrolase, a key enzyme in regulating methylations including those that are required for the maturation of viral mRNA.

Characterization of ribulose 1,5-bisphosphate carboxylase/oxygenase from Euglena gracilis Z.

An improved method was devised to purify ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with high specific activity (2.1 mumol of CO2 fixed/mg protein/min) from Euglena gracilis Z. The purified enzyme stored at -80 degrees C required treatment with dithiothreitol for full activity. The dithiothreitol-treated RuBisCO was activated by 12 mM NaHCO3 and 20 mM MgCl2, and the activated state was stable at least for 60 min in the presence of 4 mM ethylenediaminetetraacetate. The form of inorganic carbon fixed by the Euglena enzyme was CO2, as for the plant enzymes. The carboxylase reaction proceeded linearly with time for at least 8 min. The optimum pH for this reaction was 7.8 to 8.0. The carboxylase activity increased with increasing temperature up to 50 degrees C. The activation energy for the carboxylation reaction was 10.0 kcal/mol. The Michaelis constants of Euglena RuBisCO were 30.9 microM for CO2, 560 microM for O2, and 10.5 microM for ribulose 1,5-bisphosphate. Mathematical comparison between the photosynthesis rate predicted from these enzymatic properties and the observed rate suggested that there is no CO2-concentrating mechanism in E. gracilis.

Pantothenate synthetase of Escherichia coli B. I. Physicochemical properties.

Using a new apparatus for preparative polyacrylamide gel electrophoresis, pantothenate synthetase (D-pantoate: beta-alanine ligase (AMP-forming), [EC 6.3.2.1] was purified about 500-fold from Escherichia coli B. It was found to be homogeneous in analytical disc gel electrophoresis and sedimentation ultracentrifugation (so20, w=4.9). From sedimentation equilibrium ultracentrifugation, a molecular weight of 70,100 was obtained, which is in good agreement with the value obtained by the Sephadex G-150 gel filtration method (69,000); the diffusion constant was calculated to be 5.88X10(-7) cm2/sec. The minimum molecular weight calculated from the amino acid composition of this enzyme protein was 19,700, a value in reasonable accord with molecular weight of the enzyme subunit, 18,000, obtained by gel electrophoresis in the presence of sodium dodecylsulfate. The partial specific volume, v, was calculated to be 0.71 cm3/g. The enzyme had an amino-terminal glycyl residue and a Leu-Ala-Ser-OH sequence at the carboxyl end. Electrophoresis of the enzyme with carrier ampholine gave an isoelectric point of pH 4.6.

Two omega-amino acid transaminases from Bacillus cereus.

Bacillus cereus strain K-22 produced two distinct omega-amino acid transaminases, one catalyzing the transamination between beta-alanine and pyruvic acid and the other that between gamma-aminobutyric acid and alpha-ketoglutaric aic. The two enzymes were partially purified and separated from each other by various chromatographies. beta-Alanine:pyruvic acid transaminase and gamma-aminobutyric acid:alpha-ketoglutaric acid transaminase were induced by the addition of beta-alanine and gamma-aminobutyric acid, respectively, to the growth medium. beta-Alanine transaminase showed an optimum pH of 10.0 and optimum temperature of 35 degrees C, and its Km values for beta-alanine and pyruvic acid were both 1.1 mM. gamma-Aminobutyric acid, epsilon-aminocaproic acid, 2-aminoethylphosphonic acid, and propylamine showed about 30-40% of the activity of beta-alanine as amino donors, and oxalacetic acid was as good an amino acceptor as pyruvic acid. The optimum pH and temperature of gamma-aminobutyric acid transaminase were 9.0 and 50 degrees C, respectively, and its Km value for gamma-aminobutyric acid was 2.8 mM, while that for alpha-ketoglutaric acid was 2.3 mM. gamma-Aminobutyric acid and delta-aminovaleric acid were good amino donors but other omega-amino acids were virtually inactive with gamma-aminobutyric acid transaminase; alpha-ketoglutaric acid, and to a lesser extent glyoxylic acid, were active amino acceptors. Sulfhydryl reagents specifically activated gamma-aminobutyric acid transaminase.

Isolation, purification, and characterization of the pellicle of Euglena gracilis z.

The pellicle was isolated from the cell homogenate obtained on sonication of Euglena gracilis z grown aerobically under illumination and purified by a combination of differential and sucrose density gradient centrifugations. The purity and homogeneity of the pellicle fragments were determined by an electron microscopic method and biochemical analysis of the components. The protein, lipid, and sugar contents of the purified pellicle were 68.7, 17.9, and 13.5%, respectively. The equilibrium density of pellicle fragments was 1.21 g/cm3. SDS-polyacrylamide gel electrophoresis revealed that the pellicle contained 50 mol% of nonpolar amino acids. The constituents of the lipid and sugar were very different from those of the cell membrane of other organisms.

Purification and some properties of short chain-length specific trans-2-enoyl-CoA reductase in mitochondria of Euglena gracilis.

Short chain-length specific trans-2-enoyl-CoA reductase (reductase I), which contributed to mitochondrial fatty acid synthesis, was purified about 200-fold from crude extract of mitochondria in Euglena gracilis. It had a molecular weight of 39,000, and consisted of two dissimilar subunits with molecular weights of 15,000 and 25,000. The enzyme utilized crotonyl-CoA as the most active substrate and showed negative cooperativity in the reaction with the substrate. NADH was the sole electron donor. Some divalent cations were inhibitory to the enzyme when incubated with the enzyme prior to the start of the reaction. The reductase apparently contained loosely bound FAD.

Occurrence of oxygen-sensitive, NADP+-dependent pyruvate dehydrogenase in mitochondria of Euglena gracilis.

Pyruvate dehydrogenase found in mitochondria of Euglena gracilis was active on NADP+ but not NAD+, and FAD and methyl viologen also served as electron acceptors. For 2-oxoglutarate dehydrogenase both NAD+ and NADP+ were utilized and the ratio of its activity on NAD+ and NADP+ was about 1:5. The activity of pyruvate dehydrogenase was inhibited by pyruvate in aerobiosis, while not in anaerobiosis.

Mitochondrial alcohol dehydrogenase from ethanol-grown Euglena gracilis.

The inducing effects of ethanol on alcohol dehydrogenase and the key enzymes of the glyoxylate cycle, isocitrate lyase and malate synthase, in Euglena cells were investigated. Ethanol as the sole carbon source resulted in increases in alcohol dehydrogenase and the two glyoxylate cycle enzymes. The experimental results indicated that ethanol is assimilated by alcohol dehydrogenase and the glyoxylate cycle in Euglena. Mitochondria from aerobically grown Euglena contain a unique type of alcohol dehydrogenase that accounts for their ability to respire with ethanol as a substrate. This alcohol dehydrogenase was purified to homogeneity from ethanol-grown Euglena gracilis. The mitochondrial alcohol dehydrogenase was NAD(+)-specific but not NADP(+)-specific. Ethanol was the most active substrate, but the enzyme was also active towards 1-butanol, 1-heptanol, cinnamyl alcohol, and myristyl alcohol. These results indicated that mitochondrial alcohol dehydrogenase participated in alcohol metabolism in Euglena gracilis.

Epidemiology of rotavirus in Guayaquil, Ecuador.

Detection of rotavirus by electron microscopy was conducted with fecal specimens from 1,722 infants and young children with acute diarrhea, during a 41-month survey from April 1978 through December 1981 in Guayaquil, Ecuador; 376 of these specimens (21.8%) were positive. The detection rate was higher during the dry season (May to November; 25.2%) than during the rainy season (December to April; 14.7%). When rotaviruses isolated from 59 patients hospitalized with diarrhea (from April 1979 to July 1981) were subjected to genome RNA analysis by polyacrylamide gel electrophoresis, a single dominant electropherotype was found with other less common electropherotypes. An atypical rotavirus with a unique property was also found.

Effects of deicing salt on the vitality and health of two spruce species, Picea abies Karst., and Picea glehnii Masters planted along roadsides in northern Japan.

In northern Japan, the growth of Picea abies Karst., and Picea glehnii Masters, which have been planted along the highways, is often suppressed due to several environmental stresses. To examine the adverse effects of deicing salt, the primary source of stress,we measured needle life span, photosynthetic capacity, and water potential and transpiration rate of the two spruce species at a site with damaged trees, near the roadside and a site with healthy trees, located far from the highway. Results from the analysis showed large amounts of sodium and chlorine in the soil and snow at the damaged site. These elements had accumulated in the needles of the spruce. Moreover, physiological traits of the spruce, at the damaged site were also affected. Therefore, we concluded that poor physiological traits might be attributed to an accumulation of deicing salt in the needles, resulting in the suppression of tree growth.

Contents and compositions of the aroma in "Wasanbon" sugar.

"Wasanbon" sugar is handmade sugar which has been manufactured traditionally in Japan by a unique refining procedure, and is used in the making of Japanese traditional confectionary. No reports have been published on the substances responsible for the unique aroma of "Wasabon" sugar. In this paper, the contents and compositions of the aroma in "Wasabon" sugar and refinery final molasses are reported as studied by column chromatography, gas chromatography, mass spectrometry and sensory evaluation. The samples are the first press-off molasses ("Ara-mitsu" molasses) refinery final molasses, "Shiroshita" sugar (prerefined sugar) and "Wasabon" sugar. The summarized results are as follows: In the acidic fraction, the aroma of 3-phenylpropionic acid is similar to the stored aroma of "Wasabon" sugar, whereas the aroma of its methyl ester was not similar to that aroma. Although aroma contents of the weakly acidic fraction in "Wasabon" sugar and refinery final molasses are 8.5 to 8.7% of those of the acidic fraction, and their main components are cyclotene and maltol, which are formed by thermal degradation of sugar. These components show a higher preference than other weakly acidic fraction aromas, by a paired preference test. Cyclotene and maltol increased about 3.7 and 1.5 times, respectively, by the heating of "Shiroshita" sugar.

Enzymological properties of pantothenate synthetase from Escherichia coli B.

Following a previous report on physicochemical properties, the enzymological properties of a homogeneously purified preparation of pantothenate synthetase were described. The optimum pH was 10.0 and optimum temperature 30 degrees C. The lyophilized enzyme was very stable on standing at -20 degrees C. K+ or NH4+ and Mg2+ were required as activators; other cations examined were inhibitive to various extents and the enzyme required ATP as the energy supplier. Some omega-amino acids exerted strong inhibition, and the enzyme was inhibited by some chelating agents but was not affected by SH compounds and SH inhibitors. Apparent Km for pantoate was 6.3 x 10(-5)M, for beta-alanine 1.5 x 10(-4)M, and for ATP 1.0 x 10(-4)M. According to the method of Cleland, the enzyme reaction proceeds by a Bi Uni Uni Bi Ping Pong mechanism and a scheme showing the order of binding of substrates and releasing of products is presented.

The biosynthetic pathway of L-ascorbic acid in Euglena gracilis Z.

D-Glucose and D-galactose were the starting materials of L-ascorbic acid biosynthesis in Euglena gracilis as evidenced by feeding experiments of unlabeled and labeled sugars, but D-glucose was the more effective precursor. The addition of various acid derivatives of D-glucose and D-galactose, with the exception of D-glucono-delta-lactone, considerably augmented L-ascorbic acid formation. D-Galacturonic acid and L-galactono-gamma-lactone showed greater effects than did D-glucurono-gamma-lactone and L-gulono-gamma-lactone. The results of isotopic dilution experiments also showed the preference for the galacto-configuration. Fed U-14C-D-glucose was transformed into labeled D-galacturonic acid to a greater extent than into labeled D-glucuronic acid, and added D-galacturonic acid only caused extensive accumulation of labeled D-galacturonic acid. These results together show that the pathway involving D-galacturonic acid and L-galactono-gamma-lactone is the major one, the one involving D-glucuronic acid L-gulono-gamma-lactone being the minor one. A likely pathway for L-ascorbic acid biosynthesis in Euglena is proposed in the Scheme, which thus involves uronic acid intermediates and configurational inversion.