A novel orexin antagonist from a natural plant was discovered using zebrafish behavioural analysis.

OBJECTIVE: Phenotypic screening is one of the most practical approaches to the identification of mediators of behaviour, since it is difficult to model brain function in vitro, at a cellular level. We used a zebrafish (Danio rerio) behavioural assay to discover novel, natural, neuroactive compounds. MATERIALS AND METHODS: A zebrafish behavioural assay was performed for seven natural compounds, obtained from plants. The behavioural profiles were compared to those of known psychoactive drugs. We characterised a natural compound exhibiting a behaviour profile similar to that of suvorexant, using in silico, in vitro and microarray expression analysis. RESULTS: The behavioural analysis performed in this study classified central nervous system drugs according to their mechanism. Zebrafish treated with a natural compound, 8b-(4'-Hydroxytigloyloxy) costunolide (8b), showed behaviour profiles similar to those of zebrafish treated with suvorexant, a known orexin antagonist. This behavioural assay was validated using in silico and in vitro assays, which revealed that the new compound was a dual orexin receptor antagonist. In addition, transcriptome analysis suggested that 8b might regulate the nuclear factor-κB (NF-κB) related pathway. CONCLUSIONS: We conclude that zebrafish phenotypic screening, combined with in silico assays and gene expression profiling, is a useful strategy to discover and characterize novel therapeutic compounds, including natural products.

Optimization of laser-target parameters for the production of stable lithium beam.

A laser ion source coupled with a radio frequency quadrupole linac accelerator is being proposed as a suitable system for the production of a low energy, high-current stable lithium beam. In order to maximize the lithium yield, plasmas generated by laser ablation of different materials based on lithium (Li, LiOH, and LiNbO3) have been characterized by using a Faraday cup and an electrostatic ion analyzer in the time of flight configuration. A wide range of laser power density has been investigated (109-1012 W/cm2) using two Nd:YAG lasers operating at different wavelengths (1064 nm and 532 nm), pulse durations (6 ns and 17 ns), and maximum energies (1400 mJ and 210 mJ). This paper outlines the pros and cons of the investigated materials by studying how the ion energy, yields, and charge state distributions are modified when the laser power density is changed. Considerable attention has been paid to the higher charge states of oxygen, which may occur with the same mass-to-charge ratio of Li3+. The analysis has evidenced that LiNbO3 represents a valid target since it allows minimizing the O6+/7Li3+ ratio down to 2.5% by using a laser power density of 1.8 × 1010 W/cm2. For such a condition, a Li3+ current of 1.4 mA/cm2 has been measured.

Feasibility study of a compact heavy ion source for investigation of laboratory magnetospheric plasma.

We are developing a laser ion source to provide a high brightness multi-charged heavy ion beam as a part of the heavy ion beam probe system, which will be used to diagnose plasma potential in the Ring Trap 1 device at the University of Tokyo. As a probe beam, Nb2+ was selected, and a detailed laser irradiation condition was explored. It was found that the laser power density of 1.2 × 109 W/cm2 gives the maximum particle number of Nb2+ per laser energy from a niobium foil target. Essential ablation plasma parameters to design the laser ion source were also obtained. The expected beam current was more than 12 mA/cm2, with a pulse width of 3.1 µs at 200 mm away from the target.

Design of target irradiation and diagnostic chamber to study ps-laser generated plasma as a source of singly charged ions for external injection into an electron beam ion source.

High repetition-rate (∼10 kHz) ps-lasers are becoming available on the market with reasonable cost and may offer several advantages compared to ns-lasers by generating nearly continuous beams of singly charged ions appropriate for the "slow" injection mode into the Electron Beam Ion Source (EBIS). To evaluate these advantages, we will perform studies of a ps-laser generated plasma using a laser with a pulse duration of 8 ps and energy up to 5 mJ per pulse. A vacuum chamber equipped with a 3D target positioner, a focusing lens, and a Faraday Cup has been designed and built for this study. Lens-to-target distance variations have been measured using a laser tracker over the whole range of horizontal and vertical translation for all five targets we will use. The variations were found to be within ±150 µm. This degree of "target flatness" should be acceptable for our experimental conditions. Ion currents and ion pulse durations of various elements (from Al to Ta) will be measured for different target irradiation conditions (focal spot size and laser pulse energy). The results obtained will allow us to specify all parameters and geometry of a laser ion source based on a ps-laser to provide external ion injection into the relativistic heavy ion collider EBIS.

96Zr beam production for isobar experiment in relativistic heavy ion collider.

To investigate the chiral magnetic effect, 96Zr and 96Ru beams were accelerated at the relativistic heavy ion collider (RHIC) during Run-18 at Brookhaven National Laboratory. The 96Zr beam was provided from the electron beam ion source (EBIS) injector, which consists of a laser ion source, an EBIS high charge state ion breeder, a 300 keV/u radio frequency quadrupole, and a 2 MeV/u interdigital H type drift tube linear accelerator (IH-DTL). The natural abundance of 96Zr is only 2.8% with about 50% of 90Zr. To obtain a sufficient beam current, Zr material enriched to about 60% of 96Zr was used. The only available form of the enriched material was zirconium oxide (ZrO2) powder, which was not well suited for a laser ion source target. We studied and established a sintering technique of the ZrO2 powder to make a solid sample which could be installed into the laser ion source. The singly charged Zr was produced in a laser ablation plasma, extracted, and delivered to the EBIS to be ionized further to 96Zr16+. We optimized the laser irradiation condition, the EBIS confinement time, and transport through the RF linacs to maximize the performance of the injector. The total number of shots provided from the laser ion source for injection into the EBIS was 489 910. The EBIS facility provided a 192 MeV stable beam of 96Zr16+ ions to the booster ring of alternating gradient synchrotron (AGS) for further acceleration and stripping in the AGS/RHIC complex, allowing for successful data acquisition at the Solenoidal Tracker at the RHIC.

Cluster ion source for external injection and high capacity filling of light elements into the relativistic heavy ion collider electron beam ion source.

An advanced Electron Beam Ion Source (EBIS) is the primary ion source to supply highly charged ion beams of different elements to the Relativistic Heavy Ion Collider (RHIC) and to the NASA Space Radiation Laboratory (NSRL). Intense beams of highly charged ions of various elements of the periodic table, ranging from helium to uranium, have been demonstrated since EBIS became operational in 2010. EBIS routinely provides ion beams to RHIC and NSRL quasisimultaneously with about 1 s switching time between different ion species. Such unique flexibility and rapid switching between ion species are based on external injection of singly charged ions into the EBIS trap either in "fast" or "slow" injection modes. At present, a Laser Ion Source (LIS) provides most of the ion species of solid materials using the "fast" injection mode into the EBIS trap and a Hollow Cathode Ion Source (HCIS) provides most of the ion species of gaseous elements using the "slow" injection mode into the EBIS trap. Gas injection into the EBIS trap is also possible and has been used but imposes some restrictions for the simultaneous generation of highly charged ions such as Au32+ ions for RHIC and ions of gaseous species for NSRL. Because light ions have relatively high velocity inside the EBIS trap, efficient injection of hydrogen and helium ions and filling of the EBIS trap to high capacity is difficult from either LIS or HCIS. To overcome this restriction and enhance EBIS operational capability, we suggest injecting beams of hydrogen and helium cluster ions into the EBIS trap. Required parameters of cluster ion beam injection into the EBIS trap are estimated, and advantages of such an injection are highlighted. A cluster ion source with required high intensity is visible and will be designed, built, optimized, and tested.

Biological characteristics of staphylococcal enterotoxin Q and its potential risk for food poisoning.

AIMS: To elucidate the biological characteristics and stability of a newly identified staphylococcal enterotoxin Q (SEQ) against heating and digestive enzymes and to evaluate the risk of seq-harbouring Staphylococcus aureus in food poisoning. METHODS AND RESULTS: Purified SEQ was treated with heating, pepsin and trypsin which are related to food cooking, stomach and intestine conditions, respectively. Superantigenic activity of SEQ was assessed by determining the ability of IL-2 induction in mouse spleen cells. The emetic activity of SEQ was assessed using house musk shrew, a small emetic animal model. The results revealed that SEQ exhibits a remarkable resistance to heat treatment and pepsin digestion and has significant superantigenic and emetic activities. Furthermore, a sandwich ELISA for detection of SEQ production was developed, and the results showed that seq-harboring S. aureus isolates produce a large amount of SEQ. CONCLUSIONS: The newly identified SEQ had remarkable stability to heat treatment and digestive enzyme degradation and exhibited significant superantigenic and emetic activities. In addition, seq-harbouring S. aureus isolated from food poisoning outbreaks produced a large amount of SEQ, suggesting that seq-harbouring S. aureus could potentially be a hazard for food safety. SIGNIFICANCE AND IMPACT OF THE STUDY: This study found, for the first time, that SEQ, a nonclassical SE, had remarkable stability to heat treatment and enzyme degradation and exhibited significant emetic activity, indicating that SEQ is a high-risk toxin in food poisoning.

Calcium and lithium ion production for laser ion source.

Calcium and lithium ion beams are required by NASA Space Radiation Laboratory at Brookhaven National Laboratory to simulate the effects of cosmic radiation. To identify the difficulties in providing such highly reactive materials as laser targets, both species were experimentally tested. Plate shaped lithium and calcium targets were fabricated to create ablation plasmas with a 6 ns 1064 nm neodymium-doped yttrium aluminum garnet laser. We found significant oxygen contamination in both the Ca and Li high charge state beams due to the rapid oxidation of the surfaces. A large spot size, low power density laser was used to create low charge state beams without scanning the targets. The low charge state Ca beam did not have any apparent oxygen contamination, showing the potential to clean the target entirely of oxide with a low power beam once in the chamber. The Li target was clearly still oxidizing in the chamber after each low power shot. To measure the rate of oxidation, we shot the low power laser at the target repeatedly at 10 s, 30 s, 60 s, and 120 s interval lengths, showing a linear relation between the interval time and the amount of oxygen in the beam.

Proton beam production by a laser ion source with hydride target.

We studied proton beam production from a laser ion source using hydrogen rich target materials. In general, gas based species are not suitable for laser ion sources since formation of a dense laser target is difficult. In order to achieve reliable operation, we tested hydride targets using a sub nanosecond Q-switched Nd-YAG laser, which may help suppress target material consumption. We detected enough yields of protons from a titanium hydride target without degradation of beam current during the experiment. The combination of a sub nanosecond laser and compressed hydride target may provide stable proton beam.

Behavior of moving plasma in solenoidal magnetic field in a laser ion source.

In a laser ion source, a solenoidal magnetic field is useful to guide the plasma and to control the extracted beam current. However, the behavior of the plasma drifting in the magnetic field has not been well understood. Therefore, to investigate the behavior, we measured the plasma ion current and the total charge within a single pulse in the solenoid by changing the distance from the entrance of the solenoid to a detector. We observed that the decrease of the total charge along the distance became smaller as the magnetic field became larger and then the charge became almost constant with a certain magnetic flux density. The results indicate that the transverse spreading speed of the plasma decreased with increasing the field and the plasma was confined transversely with the magnetic flux density. We found that the reason of the confinement was not magnetization of ions but an influence induced by electrons.

Effect of the solenoid in various conditions of the laser ion source at Brookhaven National Laboratory.

In the laser ion source (LIS) at the Brookhaven National Laboratory (BNL), a solenoid is used to guide the laser ablation plasma and modulate the extracted beam current. Many types of ion species are guided. In some cases, the plasma plume is injected into the solenoid away from the solenoidal axis. To investigate the effects of the solenoid on the beam extracted from the plasma that has different properties, the beam current was measured in the setup of the LIS at the BNL. The beam current of Li, Al, Si, Fe, and Au increased when the magnetic field was applied. For most of the species the peak current and the total charge within a single beam pulse increased around 10 times with a magnetic field less than 100 G. In addition, for some species the rate of increase of the peak currents became smaller when the magnetic flux densities were larger than certain values depending on the species. In this case, the current waveforms were distorted. At the same magnetic field value, the field was more effective on lighter species than on heavier ones. When plasma was injected offset from the axis of the solenoid, peak current and total charge became half of those without offset. The experimental data are useful for the operation of the LIS at the BNL.

Low charge state heavy ion production with sub-nanosecond laser.

We have investigated laser ablation plasma of various species using nanosecond and sub-nanosecond lasers for both high and low charge state ion productions. We found that with sub-nanosecond laser, the generated plasma has a long tail which has low charge state ions determined by an electrostatic ion analyzer even under the laser irradiation condition for highly charged ion production. This can be caused by insufficient laser absorption in plasma plume. This property might be suitable for low charge state ion production. We used a nanosecond laser and a sub-nanosecond laser for low charge state ion production to investigate the difference of generated plasma using the Zirconium target.

Laser ion source for isobaric heavy ion collider experiment.

Heavy-ion collider experiment in isobaric system is under investigation at Relativistic Heavy Ion Collider. For this experiment, ion source is required to maximize the abundance of the intended isotope. The candidate of the experiment is (96)Ru + (96)Zr. Since the natural abundance of particular isotope is low and composition of isotope from ion source depends on the composites of the target, an isotope enriched material may be needed as a target. We studied the performance of the laser ion source required for the experiment for Zr ions.

Iron beam acceleration using direct plasma injection scheme.

A new set of vanes of radio frequency quadrupole (RFQ) accelerator was commissioned using highly charged iron beam. To supply high intensity heavy ion beams to the RFQ, direct plasma injection scheme (DPIS) with a confinement solenoid was adopted. One of the difficulties to utilize the combination of DPIS and a solenoid field is a complexity of electro magnetic field at the beam extraction region, since biasing high static electric field for ion extraction, RFQ focusing field, and the solenoid magnetic field fill the same space simultaneously. To mitigate the complexity, a newly designed magnetic field clamps were used. The intense iron beam was observed with bunched structure and the total accelerated current reached 2.5 nC.

Interaction of plasmas in laser ion source with double laser system.

Multiple laser shots could be used to elongate an ion beam pulse width or to intensify beam current from laser ion sources. In order to confirm the feasibility of the multiple shot scheme, we investigated the properties of plasmas produced by double laser shots. We found that when the interval of the laser shots is shorter than 10 µs, the ion current profile had a prominent peak, which is not observed in single laser experiments. The height of this peak was up to five times larger than that of single laser experiment.

Multiple species beam production on laser ion source for electron beam ion source in Brookhaven National Laboratory.

Extracted ion beams from the test laser ion source (LIS) were transported through a test beam transport line which is almost identical to the actual primary beam transport in the current electron beam ion source apparatus. The tested species were C, Al, Si, Cr, Fe, Cu, Ag, Ta, and Au. The all measured beam currents fulfilled the requirements. However, in the case of light mass ions, the recorded emittance shapes have larger aberrations and the RMS values are higher than 0.06 π mm mrad, which is the design goal. Since we have margin to enhance the beam current, if we then allow some beam losses at the injection point, the number of the single charged ions within the acceptance can be supplied. For heaver ions like Ag, Ta, and Au, the LIS showed very good performance.

Comparison of graphite materials for targets of laser ion source.

To investigate efficient graphite material for carbon ion production in laser ion source, the plasma properties produced from these materials are measured. Comparing acquired current profile and charge state distribution, the distributions of ions in laser induced plasma from isotropic graphite and single crystal of graphite are different. The produced quantity of C(6+) from isotropic materials is larger than that from single crystal.

Immunization with outer membrane protein A from Salmonella enterica serovar Enteritidis induces humoral immune response but no protection against homologous challenge in chickens.

Vaccination of poultry is one promising strategy to mitigate Salmonella infection in poultry and, in turn, humans as well. We evaluated the efficacy of outer membrane protein A (OmpA) as a novel vaccine candidate against Salmonella in poultry. Native OmpA purified from Salmonella enterica serovar Enteritidis was mixed with adjuvant and administered intramuscularly to 41-d-old chicks. The vaccinated birds showed no decrease in cecal excretion and tissue colonization compared with the unvaccinated birds after oral challenge with 10(9) cfu of the homologous strain at 28 d postimmunization. However, this vaccination induced an increased level of serum anti-OmpA IgG. Similar results were obtained in the replication experiments using a recombinant OmpA with single and double doses. For the development of more effective component vaccines for avian salmonellosis, the vaccine efficacy of outer membrane proteins other than OmpA and route of immunization other than parenteral administration should be evaluated with regard to protection and immune responses, including mucosal IgA.