Structure and orientation of antibiotic peptide alamethicin in phospholipid bilayers as revealed by chemical shift oscillation analysis of solid state nuclear magnetic resonance and molecular dynamics simulation.

The structure, topology and orientation of membrane-bound antibiotic alamethicin were studied using solid state nuclear magnetic resonance (NMR) spectroscopy. (13)C chemical shift interaction was observed in [1-(13)C]-labeled alamethicin. The isotropic chemical shift values indicated that alamethicin forms a helical structure in the entire region. The chemical shift anisotropy of the carbonyl carbon of isotopically labeled alamethicin was also analyzed with the assumption that alamethicin molecules rotate rapidly about the bilayer normal of the phospholipid bilayers. It is considered that the adjacent peptide planes form an angle of 100° or 120° when it forms α-helix or 310-helix, respectively. These properties lead to an oscillation of the chemical shift anisotropy with respect to the phase angle of the peptide plane. Anisotropic data were acquired for the 4 and 7 sites of the N- and C-termini, respectively. The results indicated that the helical axes for the N- and C-termini were tilted 17° and 32° to the bilayer normal, respectively. The chemical shift oscillation curves indicate that the N- and C-termini form the α-helix and 310-helix, respectively. The C-terminal 310-helix of alamethicin in the bilayer was experimentally observed and the unique bending structure of alamethicin was further confirmed by measuring the internuclear distances of [1-(13)C] and [(15)N] doubly-labeled alamethicin. Molecular dynamics simulation of alamethicin embedded into dimyristoyl phophatidylcholine (DMPC) bilayers indicates that the helical axes for α-helical N- and 310-helical C-termini are tilted 12° and 32° to the bilayer normal, respectively, which is in good agreement with the solid state NMR results.

Consumption of tyrosine in royal jelly increases brain levels of dopamine and tyramine and promotes transition from normal to reproductive workers in queenless honey bee colonies.

Dopamine (DA) and tyramine (TA) have neurohormonal roles in the production of reproductive workers in queenless colonies of honey bees, but the regulation of these biogenic amines in the brain are still largely unclear. Nutrition is an important factor in promoting reproduction and might be involved in the regulation of these biogenic amines in the brain. To test this hypothesis, we examined the effect of oral treatments of tyrosine (Tyr; a common precursor of DA, TA and octopamine, and a component of royal jelly) in queenless workers and quantified the resulting production of biogenic amines. Tyrosine treatments enhanced the levels of DA, TA and their metabolites in the brain. Workers fed royal jelly had significantly larger brain levels of Tyr, DA, TA and the metabolites in the brains compared with those bees fed honey or sucrose (control). Treatment with Tyr also inhibited the behavior of workers outside of the hive and promoted ovarian development. These results suggest that there is a link between nutrition and the regulation of DA and TA in the brain to promote the production of reproductive workers in queenless honey bee colonies.

Method for validating cloud mask obtained from satellite measurements using ground-based sky camera.

Error propagation in Earth's atmospheric, oceanic, and land surface parameters of the satellite products caused by misclassification of the cloud mask is a critical issue for improving the accuracy of satellite products. Thus, characterizing the accuracy of the cloud mask is important for investigating the influence of the cloud mask on satellite products. In this study, we proposed a method for validating multiwavelength satellite data derived cloud masks using ground-based sky camera (GSC) data. First, a cloud cover algorithm for GSC data has been developed using sky index and bright index. Then, Moderate Resolution Imaging Spectroradiometer (MODIS) satellite data derived cloud masks by two cloud-screening algorithms (i.e., MOD35 and CLAUDIA) were validated using the GSC cloud mask. The results indicate that MOD35 is likely to classify ambiguous pixels as "cloudy," whereas CLAUDIA is likely to classify them as "clear." Furthermore, the influence of error propagations caused by misclassification of the MOD35 and CLAUDIA cloud masks on MODIS derived reflectance, brightness temperature, and normalized difference vegetation index (NDVI) in clear and cloudy pixels was investigated using sky camera data. It shows that the influence of the error propagation by the MOD35 cloud mask on the MODIS derived monthly mean reflectance, brightness temperature, and NDVI for clear pixels is significantly smaller than for the CLAUDIA cloud mask; the influence of the error propagation by the CLAUDIA cloud mask on MODIS derived monthly mean cloud products for cloudy pixels is significantly smaller than that by the MOD35 cloud mask.

Juvenile hormone-dopamine systems for the promotion of flight activity in males of the large carpenter bee Xylocopa appendiculata.

The reproductive roles of dopamine and dopamine regulation systems are known in social hymenopterans, but the knowledge on the regulation systems in solitary species is still needed. To test the possibility that juvenile hormone (JH) and brain dopamine interact to trigger territorial flight behavior in males of a solitary bee species, the effects on biogenic amines of JH analog treatments and behavioral assays with dopamine injections in males of the large carpenter bee Xylocopa appendiculata were quantified. Brain dopamine levels were significantly higher in methoprene-treated males than in control males 4 days after treatment, but were not significantly different after 7 days. Brain octopamine and serotonin levels did not differ between methoprene-treated and control males at 4 and 7 days after treatment. Injection of dopamine caused significantly higher locomotor activities and a shorter duration for flight initiation in experimental versus control males. These results suggest that brain dopamine can be regulated by JH and enhances flight activities in males. The JH-dopamine system in males of this solitary bee species is similar to that of males of the highly eusocial honeybee Apis mellifera.

Neuroendocrine mechanisms underlying regulation of mating flight behaviors in male honey bees (Apis mellifera L.).

We determined the neuroendocrine mechanisms underlying regulation of mating flight behaviors in male honey bees. Both a precursor of dopamine (3,4-dihydroxyphenylalanine: DOPA) and a precursor of octopamine (tyramine) in the brain decreased in an age-dependent fashion before sexual maturation (i.e. 8days of age), whereas the levels of brain dopamine, dopamine metabolites (N-acetyldopamine and norepinephrine) and octopamine were increased. These age-dependent increases of dopamine and octopamine were also detected in the meso-metathoracic ganglia. Injection of either dopamine or octopamine into 7-8-day-old males shortened the duration for flight-initiation and increased the duration of wing vibration, indicating that both dopamine and octopamine enhance the flight-initiation and -sustaining activities in males. Applications of a juvenile hormone analog (methoprene) enhanced the levels of dopamine in the brains of 4-day-old males, but this enhancement was not detected in either brain octopamine or meso-metathoracic dopamine and octopamine. Thus, we found that both dopamine and octopamine in the brain and meso-metathoracic ganglia increase until sexual maturation and could enhance the activities of mating flight independently; in addition, the increase in levels of dopamine in the brain could be selectively regulated by juvenile hormone. The regulatory systems of dopamine and octopamine in honey bee males might be 'classical' and similar to those of primitively eusocial hymenopterans, and partly adapt to the short lifespan with a single mating system in the males.

Superflares on solar-type stars.

Solar flares are caused by the sudden release of magnetic energy stored near sunspots. They release 10(29) to 10(32) ergs of energy on a timescale of hours. Similar flares have been observed on many stars, with larger 'superflares' seen on a variety of stars, some of which are rapidly rotating and some of which are of ordinary solar type. The small number of superflares observed on solar-type stars has hitherto precluded a detailed study of them. Here we report observations of 365 superflares, including some from slowly rotating solar-type stars, from about 83,000 stars observed over 120 days. Quasi-periodic brightness modulations observed in the solar-type stars suggest that they have much larger starspots than does the Sun. The maximum energy of the flare is not correlated with the stellar rotation period, but the data suggest that superflares occur more frequently on rapidly rotating stars. It has been proposed that hot Jupiters may be important in the generation of superflares on solar-type stars, but none have been discovered around the stars that we have studied, indicating that hot Jupiters associated with superflares are rare.

Caste differences in dopamine-related substances and dopamine supply in the brains of honeybees (Apis mellifera L.).

We determined the mechanisms underlying caste differences in the brain levels of dopamine-related substances in adult honeybees (Apis mellifera L.). Brain levels of dopamine, DOPA (a dopamine precursor), and N-acetyldopamine (a dopamine metabolite, NADA) were significantly higher in three-day-old virgin queens than same-aged workers. Caste differences in dopamine and NADA levels were also found in the hemolymph. The in vitro enzymatic activities of DOPA decarboxylase (DDC) during dopamine synthesis in brains were not significantly different between castes. The DDC activity in adult queens was mainly found in the brain, but with lower levels of activity detected in the mandibular glands, salivary glands and ovaries. Oral application of DOPA to workers led to DOPA uptake in the brain and significantly higher dopamine and NADA levels in the brain, suggesting that dopamine synthesis could be controlled by the amount of DDC substrate. Royal jelly samples taken from queen cells had a >25-fold higher concentration of dopamine compared with honey samples collected from honey cells. However, oral application of the same concentration of dopamine did not significantly enhance the brain levels of dopamine and NADA. These results suggest that the higher levels of brain dopamine in queens compared with workers can be explained by the higher level of DDC substrate, rather than DDC activity in the brain and other tissues of queens as well as exogenous dopamine in the royal jelly.

Dopamine enhances locomotor activity for mating in male honeybees (Apis mellifera L.).

Dopamine plays multiple roles in the regulation of reproduction in female honeybees where it appears to act independently of juvenile hormone (JH). In males the role of dopamine and its relationship to JH control have not been elucidated. In the present study we determined hemolymph levels of dopamine and its metabolite (N-acetyldopamine) in males at post-emergence days 0-16. The development of locomotor and flight activities were recorded over the same period. Hemolymph levels of dopamine and N-acetyldopamine were found to increase at the time of onset of mating flight activity and those of dopamine decreased thereafter. Both locomotor and flight activities increased in parallel with hemolymph dopamine levels but the increased activity levels were maintained following decline of dopamine levels. Brain and meso-metathoracic ganglia levels of dopamine showed a similar developmental profile to hemolymph dopamine levels. Locomotor activities were temporarily inhibited by injection of a dopamine-receptor antagonist (cis(Z)-flupenthixol) into the thorax, and were enhanced by injection of a dopamine-receptor agonist (6,7-ADTN). These results suggest that dopamine regulates locomotor activities for mating and plays a role downstream of JH in premature males in honeybees.

Gonadotropic effects of dopamine in isolated workers of the primitively eusocial wasp, Polistes chinensis.

In social insects, biogenic amines are thought to play regulatory roles in the transition between reproductive states in females. To determine the effect of dopamine on the reproductive development of workers in primitively eusocial societies, isolated workers of the paper wasp Polistes chinensis were supplied with oral dopamine. Ovarian development was accelerated in dopamine-fed workers as compared to control workers of the same age fed only sucrose solution. Oral dopamine increased brain levels of dopamine and its metabolite (N-acetyldopamine). Brain levels of tyramine or octopamine were also increased by dopamine application in one of two colonies; levels of the tyramine metabolite N-acetyltyramine were unchanged. These results indicate that dopamine plays a gonadotropic role in isolated workers in the primitively eusocial wasp, similar to the gonadotropic role previously reported for juvenile hormone. This is the first study to report effects of dopamine on ovarian development in workers of the paper wasp.

Prognostic factors in influenza-associated encephalopathy.

BACKGROUND: Recently, reports of influenza-associated encephalopathy have increased worldwide. Given the high mortality and morbidity rates attributable to this severe neurologic complication of influenza, we conducted a nationwide study in Japan to identify the prognostic factors. METHODS: We retrospectively evaluated 442 cases of influenza-associated encephalopathy that were reported to the Collaborative Study Group on Influenza-Associated Encephalopathy, which was organized by the Japanese Ministry of Health, Labor, and Welfare in collaboration with hospitals, clinics, and local pediatric practices in Japan between 1998 and 2002. The outcome for each patient was classified as either survival or death. Predictors of death were identified using logistic regression analysis. RESULTS: Four major prognostic factors for death were found to be significant by multivariate analysis (P < 0.05) in the 184 patients for whom we had complete data: elevation of aspartate aminotransferase, hyperglycemia, the presence of hematuria or proteinuria, and use of diclofenac sodium. CONCLUSIONS: We identified patients who had factors associated with a poor prognosis, and these findings might be clinically useful for the management of this illness.

Influence of age and juvenile hormone on brain dopamine level in male honeybee (Apis mellifera): association with reproductive maturation.

Dopamine (DA) is a major functional biogenic amine in insects and has been suggested to regulate reproduction in female honeybees. However, its function has not been investigated in male drones. To clarify developmental changes of DA in drones, brain DA levels were investigated at various ages and showed a similar pattern to the previously reported juvenile hormone (JH) hemolymph titer. The DA level was lowest at emergence and peaked at day 7 or 8, followed by decline. Application of JH analog increased brain DA levels in young drones (2-4-days-old), suggesting regulation of DA by JH in drones. In young drones, maturation of male reproductive organs closely matched the increase in brain DA. The dry weight of testes decreased and that of seminal vesicles increased from emergence to day 8. The dry weight of mucus glands increased up to day 4. Consequently, DA regulated by JH might have reproductive behavior and/or physiological functions in drones.

Neuro-endocrine correlates of ovarian development and egg-laying behaviors in the primitively eusocial wasp (Polistes chinensis).

To explore the roles of biogenic amines in reproduction in workers in primitively eusocial societies, correlations between brain levels of biogenic amines and ovarian development or oviposition in workers of the paper wasp Polistes chinensis were investigated. Several workers in queenright colonies developed ovaries and were potential egg-layers. Maximum ovarian width was significantly correlated with brain levels of dopamine, serotonin and their metabolites (N-acetyldopamine and N-acetylserotonin). Individuals with developed ovaries proceeded with yolk formation had significantly higher levels of brain dopamine, serotonin and N-acetyldopamine compared with individuals with undeveloped ovaries. Brain dopamine levels were higher in egg-laying individuals than in other individuals with developed ovaries. Thus, the workers of the paper wasp showed quantitative differences in brain dopamine levels correlated with reproduction. These results suggest that the brain levels of biogenic amines in paper wasp workers correspond to their tasks, and that there is a mechanism for promoting reproduction by dopamine, as previously reported in the workers of eusocial bees.

Glutamate release from astrocytes is stimulated via the appearance of exocytosis during cyclic AMP-induced morphologic changes.

Recent studies have shown that astrocytes release various transmitters including glutamate and thus directly affect synaptic neurotransmission. The mechanisms involved in the release of glutamate from astrocytes remain unclear, however. In the present study, we examined differences in 1) the amount of glutamate released, 2) the appearance of exocytosis, and 3) the expression of SNARE (soluble N-ethylmaleimide sensitive fusion protein attachment protein receptor) proteins between cyclic AMP-treated and non-treated astrocytes in culture. Extracellular glutamate was detected in the recording solution of cyclic AMP-treated astrocytes after stimulation with ATP by high-performance liquid chromatography and NADH imaging. Exocytosis, which was observed by FM1-43 imaging, appeared in cyclic AMP-treated astrocytes in a punctiform fashion, but not in non-treated cells, after stimulation with ATP and glutamate. Immunocytochemistry and Western blotting showed that the amount of SNARE proteins increased during cAMP-induced morphologic changes, and in particular, a v-SNARE, synaptobrevin, appeared as punctiform staining in the cytosol of cyclic AMP-treated astrocytes. These findings show that astrocytes acquire SNARE proteins during cyclic AMP-induced differentiation, and suggest that glutamate is released by exocytosis in cyclic AMP-treated astrocytes in response to ATP released from neighboring neurons and astrocytes.

Experiential effects of appetitive and nonappetitive odors on feeding behavior in the blowfly, Phormia regina: a putative role for tyramine in appetite regulation.

In humans, appetite is affected by food experiences and food flavors. In the blowfly Phormia regina, we found that feeding threshold to sugar increased in the presence of the odor of D-limonene and decreased in the presence of the odor of dithiothreitol (DTT). Using these odors as representative nonappetitive and appetitive flavors, we demonstrated the role played by tyramine (TA) in appetite regulation by experiences of food flavors. When fed with sucrose flavored with D-limonene for 5 d after emergence, flies showed subsequent decreased appetite to plain sucrose, whereas when they were fed with sucrose flavored by DTT they showed increased appetite. However, mushroom body (MB)-ablated flies did not show these patterns. This suggests that MB, one of the primary memory centers of the insect brain, is necessary for the flies to apply previous experiences of food flavors to appetitive learning behaviors. In addition, flies' previously acquired decreased or increased appetites showed parallel changes with both octopamine (OA) and tyramine levels in the brain. However, injection experiments with OA, TA, or their agonist and antagonist indicated that TA more directly mediates feeding threshold determination, which was affected by acquired memories of food flavors.

Morphological behavior of lipid bilayers induced by melittin near the phase transition temperature.

Morphological changes of DMPC, DLPC, and DPPC bilayers containing melittin (lecithin/melittin molar ratio of 10:1) around the gel-to-liquid crystalline phase transition temperatures (Tc) were examined by a variety of biophysical methods. First, giant vesicles with the diameters of approximately 20 microm were observed by optical microscopy for melittin-DMPC bilayers at 27.9 degrees C. When the temperature was lowered to 24.9 degrees C (Tc = 23 degrees C for the neat DMPC bilayers), the surface of vesicles became blurred and dynamic pore formation was visible in the microscopic picture taken at different exposure times. Phase separation and association of melittin molecules in the bilayers were further detected by fluorescent microscopy and mass spectrometry, respectively. These vesicles disappeared completely at 22.9 degrees C. It was thus found that the melittin-lecithin bilayers reversibly undergo their fusion and disruption near the respective Tcs. The fluctuation of lipids is, therefore, responsible for the membrane fusion above the Tc, and the association of melittin molecules causes membrane fragmentation below the Tc. Subsequent magnetic alignments were observed by solid-state (31)P NMR spectra for the melittin-lecithin vesicles at a temperature above the respective Tcs. On the other hand, additional large amplitude motion induced by melittin at a temperature near the Tc breaks down the magnetic alignment.

Depression of brain dopamine and its metabolite after mating in European honeybee (Apis mellifera) queens.

To explore neuro-endocrinal changes in the brain of European honeybee (Apis mellifera) queens before and after mating, we measured the amount of several biogenic amines, including dopamine and its metabolite in the brain of 6- and 12-day-old virgins and 12-day-old mated queens. Twelve-day-old mated queens showed significantly lower amounts of dopamine and its metabolite (N-acetyldopamine) than both 6- and 12-day-old virgin queens, whereas significant differences in the amounts of these amines were not detected between 6- and 12-day-old virgin queens. These results are explained by down-regulation of both synthesis and secretion of brain dopamine after mating. It is speculated that higher amounts of brain dopamine in virgin queens might be involved in activation of ovarian follicles arrested in previtellogenic stages, as well as regulation of their characteristic behaviors.

Serotonin modifies the sensitivity of the male silkmoth to pheromone.

Serotonin is known to modulate the response of neuronal populations in the primary olfactory center of the moth olfactory system, the antennal lobe. Here, we analyzed the effects of serotonin on the behavior related to the restricted pheromone olfactory pathway of the male silkmoth, Bombyx mori. In order to understand the effects of serotonin at the behavioral level, we applied serotonin (10(-5) mol l(-1), 10(-4) mol l(-1) and 10(-3) mol l(-1)) to the brain and found that 10(-4) mol l(-1) serotonin increases the sensitivity to female pheromone whereas 10(-3) mol l(-1) serotonin had the opposite effect. Levels of serotonin in the brain were determined using HPLC with electrochemical detection. Inhibitory effects were observed after applying the serotonin antagonists mianserin (10(-4) mol l(-1)) and ketanserin (10(-3) mol l(-1)). Additionally, we quantified the circadian variation of serotonin in the brain using HPLC with electrochemical detection. Further, this variation correlated well with a circadian variation of the male sensitivity to pheromone. These results show that the serotonin-related enhancement of neuronal responses at the antennal lobe level is expressed at the behavioral level as a modulation of pheromone sensitivity and that the circadian variation of serotonin levels in the brain correlates with changes in the moth's pheromone sensitivity.

Dynorphin induced magnetic ordering in lipid bilayers as studied by (31)P NMR spectroscopy.

Lipid bilayers of dimyristoyl phosphatidylcholine (DMPC) containing opioid peptide dynorphin A(1-17) are found to be spontaneously aligned to the applied magnetic field near at the phase transition temperature between the gel and liquid crystalline states (T(m)=24 degrees C), as examined by 31P NMR spectroscopy. The specific interaction between the peptide and lipid bilayer leading to this property was also examined by optical microscopy, light scattering, and potassium ion-selective electrode, together with a comparative study on dynorphin A(1-13). A substantial change in the light scattering intensity was noted for DMPC containing dynorphin A(1-17) near at T(m) but not for the system containing A(1-13). Besides, reversible change in morphology of bilayer, from small lipid particles to large vesicles, was observed by optical microscope at T(m). These results indicate that lysis and fusion of the lipid bilayers are induced by the presence of dynorphin A(1-17). It turned out that the bilayers are spontaneously aligned to the magnetic field above T(m) in parallel with the bilayer surface, because a single 31P NMR signal appeared at the perpendicular position of the 31P chemical shift tensor. In contrast, no such magnetic ordering was noted for DMPC bilayers containing dynorphin A(1-13). It was proved that DMPC bilayer in the presence of dynorphin A(1-17) forms vesicles above T(m), because leakage of potassium ion from the lipid bilayers was observed by potassium ion-selective electrode after adding Triton X-100. It is concluded that DMPC bilayer consists of elongated vesicles with the long axis parallel to the magnetic field, together with the data of microscopic observation of cylindrical shape of the vesicles. Further, the long axis is found to be at least five times longer than the short axis of the elongated vesicles in view of simulated 31P NMR lineshape.