Using an Automated 3D-tracking System to Record Individual and Shoals of Adult Zebrafish

Like many aquatic animals, zebrafish (Danio rerio) moves in a 3D space. It is thus preferable to use a 3D recording system to study its behavior. The ...

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Aspergillus fumigatus is a common cause of invasive and allergic pulmonary disease. Resting conidia of the filamentous fungus are constantly inhaled, but cause infection only after initiating hyphal growth. In this study, we have explored whether macrophages can distinguish between resting spores and the maturing, potentially invasive form of the fungus. Although macrophages bind and ingest A. fumigatus resting conidia efficiently, there is little inflammatory response; NF-kappabeta is not activated, inflammatory cytokines are not induced, and reactive oxygen species are not produced. However, maturing A. fumigatus conidia and germ tubes stimulate NF-kappabeta, secretion of proinflammatory cytokines and production of reactive oxygen by human monocyte-derived macrophages and murine macrophages from multiple anatomical sites. These responses are in part mediated by dectin-1, which binds cell wall beta-glucan that is not present on the surface of dormant conidia, but is present after cellular swelling and loss of the hydrophobic proteinaceous cell wall. Dectin-1 binding to germ tubes augments, but is not required for, TLR2-mediated inflammatory cytokine secretion. Dectin-1 recognition of germ tubes also stimulates TNF-alpha production in the absence of both TLR2 and MyD88 signaling. These data demonstrate one mechanism by which the pulmonary inflammatory response is tailored toward metabolically active cells, thereby avoiding unnecessary tissue damage with frequent inhalation of ubiquitous spores.

Dectin-1 and TLRs permit macrophages to distinguish between different Aspergillus fumigatus cellular states.

The protozoan parasite Toxoplasma gondii is the causative agent of one of the most widespread parasitic infections of man and is a leading cause of congenital neurological birth defects and the third most common cause of food-borne deaths in the United States. Despite this, to date no drugs are available that provide a fully effective treatment. Recently, the antibacterial agent triclosan was shown to inhibit the fatty-acid biosynthesis pathway in T. gondii and to interact with the enoyl reductase (ENR). In order to analyse the potential of triclosan as a lead compound targeting T. gondii ENR and to explore unique features of the apicomplexan enzyme that could be exploited in future drug development, structural studies have been initiated on T. gondii ENR. Crystals of T. gondii ENR in complex with NAD+ and triclosan were grown using the hanging-drop vapour-diffusion method with PEG 8000 as precipitant. The crystals belong to space group P3(2)21, with approximate unit-cell parameters a = 78.1, b = 78.1, c = 188.5 A, alpha = beta = 90, gamma = 120 degrees and a dimer in the asymmetric unit. Test data were collected to beyond 2.6 A on cryocooled crystals (100 K) using a Rigaku MM007 rotating-anode X-ray source, revealing that the crystals are suitable for a full structural determination.

Expression, purification and preliminary crystallographic analysis of the Toxoplasma gondii enoyl reductase.

Lipoate is an essential cofactor for key enzymes of oxidative metabolism. Plasmodium falciparum possesses genes for lipoate biosynthesis and scavenging, but it is not known if these pathways are functional, nor what their relative contribution to the survival of intraerythrocytic parasites might be. We detected in parasite extracts four lipoylated proteins, one of which cross-reacted with antibodies against the E2 subunit of apicoplast-localized pyruvate dehydrogenase (PDH). Two highly divergent parasite lipoate ligase A homologues (LplA), LipL1 (previously identified as LplA) and LipL2, restored lipoate scavenging in lipoylation-deficient bacteria, indicating that Plasmodium has functional lipoate-scavenging enzymes. Accordingly, intraerythrocytic parasites scavenged radiolabelled lipoate and incorporated it into three proteins likely to be mitochondrial. Scavenged lipoate was not attached to the PDH E2 subunit, implying that lipoate scavenging drives mitochondrial lipoylation, while apicoplast lipoylation relies on biosynthesis. The lipoate analogue 8-bromo-octanoate inhibited LipL1 activity and arrested P. falciparum in vitro growth, decreasing the incorporation of radiolabelled lipoate into parasite proteins. Furthermore, growth inhibition was prevented by lipoate addition in the medium. These results are consistent with 8-bromo-octanoate specifically interfering with lipoate scavenging. Our study suggests that lipoate metabolic pathways are not redundant, and that lipoate scavenging is critical for Plasmodium intraerythrocytic survival.

Scavenging of the cofactor lipoate is essential for the survival of the malaria parasite Plasmodium falciparum.

Fish species have been used as vertebrate model systems for numerous human diseases. However, a comprehensive monitoring system for continuously tracking the positions of aquatic animals is still lacking. Manual or simple automatic methods usually lead to false or incomplete behavioral parameters. In this paper, a video-based 3D system is proposed for monitoring aquatic animals. This system is comprised of a novel catadioptric stereo-vision setup and methods for robust tracking of 3D motion-related behavior. The system has many advantages over 2D monitoring methods, such as being able to completely monitor the animals' behavior in 3D space with high spatial and temporal resolution, track multiple animals simultaneously without any physical marker, and accurately reconstruct 3D motion trajectories despite the potential problems of water refraction and reflection. Alterations in swimming behavior following exposure to acute ethanol were studied in goldfish (Carassius auratus) using the 3D behavior monitoring system. In analyzing the experimental data, a systematic comparison was made between the 3D goldfish behavioral parameters and their dimensionally reduced 2D forms. It was found that the 3D monitoring method was able to generate more accurate behavioral parameters than the conventional 2D methods. Compared to the results from 3D method, the hypothesis test conclusions based on 2D methods are more prone to error. It is expected that the 3D behavior monitoring system can significantly improve the efficiency, accuracy, and cost-effectiveness of behavioral studies and of model development in aquatic animals; potential applications include pre-clinical drug development, in-vivo compound screening, and bio-sensing.

Catadioptric stereo-vision system for the real-time monitoring of 3D behavior in aquatic animals.

This work is devoted to an extensive study of cerium deposits distributed directly on zinc particles by simple impregnation or ultrasonic impregnation for the modification of zinc powder. Meantime, the characterization of modified zinc powder and the influence of ultrasound parameters in the modification process upon the dendritic growth, the corrosion behavior and the cyclic performance of zinc are investigated using scanning electron microscopy, energy dispersion spectrometry, potentiostatic polarization, potentiodynamic polarization and cyclic voltammetry. Compared with simple impregnation, the assistance of ultrasonic irradiation is found to have a significant effect on the sedimentary state and favorable properties of cerium deposits in a protective way. Besides the cyclic voltammetry measurements display that the application of ultrasound also improves the cyclic performance of zinc electrode containing modified zinc powder mainly because the cerium deposits formed under ultrasonic irradiation can greatly hinder the dissolution and diffusion of the oxidation product of zinc in the electrolyte and effectively favor the capacity maintenance of zinc electrode.

A novel method for the modification of zinc powder by ultrasonic impregnation in cerium nitrate solution.

The levonorgestrel intrauterine system (LNG-IUS) is a widely recognized intrauterine anti-fertility system, which can alleviate symptoms of uterine leiomyoma. This study aims to evaluate leimyoma cell growth inhibition induced by high concentrations of LNG.

Levonorgestrel inhibits proliferation and induces apoptosis in uterine leiomyoma cells.

Wire-shaped quantum dots-sensitized solar cells (WS-QDSCs) based on nanosheets and nanowires were fabricated and investigated for this paper. The nanosheets grown on stainless steel (SS) wire by electrodeposition were mainly composed of Zn₅(OH)₈Cl₂·H₂O and most of the Zn₅(OH)₈Cl₂·H₂O was converted to ZnO by post-treatment, and ZnO nanowires were directly grown on SS wire by the hydrothermal method. CdS QDs were deposited on nanosheets and nanowires by successive ionic layer adsorption and reaction method. The results of photoelectrochemical performance indicated that WS-QDSCs showed a similar conversion efficiency in polysulfide and Na₂SO₄ electrolytes, while the WS-QDSCs based on the Cu2S counter electrode achieved much higher performance than those based on SS and Cu counter electrodes. By optimizing electrodeposition duration, the WS-QDSCs based on nanosheets presented the highest conversion efficiency of 0.60% for the duration of 20 min. Performance comparison indicated that the WS-QDSC based on nanosheets showed very superior performance to that based on the nanowires with similar film thickness.

Wire-shaped quantum dots-sensitized solar cells based on nanosheets and nanowires.

Air and soil pollution from traffic has been considered as a critical issue to crop production and food safety, however, few efforts have been paid on distinguish the source origin of traffic-related contaminants in rice plant along highway. Therefore, we investigated metals (Pb, Cd, Cr, Zn and Cu) concentrations and stable Pb isotope ratios in rice plants exposed and unexposed to highway traffic pollution in Eastern China in 2008. Significant differences in metals concentrations between the exposed and unexposed plants existed in leaf for Pb, Cd and Zn, in stem only for Zn, and in grain for Pb and Cd. About 46% of Pb and 41% of Cd in the grain were attributed to the foliar uptake from atmosphere, and there were no obvious contribution of atmosphere to the accumulations of Cr, Zn and Cu in grain. Except for Zn, all of the heavy metals in stem were attributed to the root uptake from soil, although significant accumulations of Pb and Cd from atmosphere existed in leaf. This indicated that different processes existed in the subsequent translocation of foliar-absorbed heavy metals between rice organs. The distinct separation of stable Pb isotope ratios among rice grain, leaf, stem, soil and vehicle exhaust further provided evidences on the different pathways of heavy metal accumulation in rice plant. These results suggested that further more attentions should be paid to the atmospheric deposition of heavy metals from traffic emission when plan crop layout for food safety along highway.

Source attributions of heavy metals in rice plant along highway in Eastern China.

One-dimensional flexible solar cells were fabricated through vertical growth of ZnO nanowires on freestanding carbon fibers and subsequent deposition of CdS quantum dots (QDs). Under light illumination, excitons were generated in the CdS QDs and dissociated in the ZnO/CdS interface. Photoelectrochemical characterization indicates that fiber quantum dot-sensitized solar cells (QDSCs) could effectively absorb visible light and convert it to electric energy. The photoelectrochemical performance was enhanced after the deposition of a ZnS passivating layer on the CF/ZnO/CdS surface. The highest conversion efficiency of about 0.006% was achieved by the fiber QDSCs. A higher conversion efficiency was expected to be achieved after some important parameters and cell structure were optimized and improved.

Growth of ZnO nanowires on fibers for one-dimensional flexible quantum dot-sensitized solar cells.

Behavioral changes in zebrafish induced by acute administration of buspirone have been interpreted to be the result of reduced anxiety. The purpose of the current study was to determine whether the effects of short-term and mid-term habituation to an open field corroborated the anxiolytic hypothesis. We exposed single zebrafish for 60 min to 5 mgL buspirone and tested them twice, immediately after exposure and again 3.5 h later. Each session lasted 20 min. Distance from bottom of tank, velocity, duration freezing, distance from center and horizontal distribution, and preferred spatial location were analyzed with a 3-D tracking system. In the early session (starting at 10:30), 20 min habituation of control zebrafish only marginally increased distance from bottom, which was still 90% higher in zebrafish treated with buspirone. When extending the habituation to 3.5 h, the picture became more complex. Distance from bottom did not further increase in control zebrafish. More importantly, some signs of increased anxiety were present in the buspirone group, such as increased freezing, reduced velocity, and increased bottom-dwelling. However, analyzing data of individual fish excluded rebound anxiety as unlikely. The delayed effects might be drug side effects, such as motor impairment and/or dizziness. The immediate and the delayed effects of buspirone have the appearance to be unrelated.

The immediate and the delayed effects of buspirone on zebrafish (Danio rerio) in an open field test: a 3-D approach.

Changing the sheets: Hollow TiO₂ porous nanosheets (HTPNs) are prepared on ITO glass through an improved TiO₂ polycrystalline shell-assisted cation exchange by using ZnO porous nanosheets (grown by using indirect electrodeposition) as a template. Quantum dot-sensitized solar cells based on HTPNs exhibit a high performance, and other photoelectrochemical devices are expected to be prepared based on HTPNs.

Hollow TiO₂ porous nanosheets: transformation from ZnO porous nanosheets and application in photoelectrochemical cells.

Understanding the different patterns of anxiety-like behavioral responses is of great interest for pharmacological and genetic research. Here we report the effects of 3.5-hr habituation, buspirone and ethanol on those responses in shoaling zebrafish (Danio rerio). Since in these experiments we used a container with white walls, the effects of black-vs.-white walls were tested in a separate experiment. An important objective was to determine whether factors unrelated to anxiety played a role in modulating the responses. The anxiety-like behavioral responses studied here are social cohesion, distance from bottom and bottom-dwell time, radial distribution (to study thigmotaxis), transparent-wall preference (to study escape responses), locomotion and freezing. The experimental conditions yielded distinctly different response patterns. Thigmotaxis was the most obvious response to white walls and it was significantly reduced after 3.5-hr habituation. It was not affected by any of the drugs. The reduction of social cohesion after 3.5-hr habituation and in the 0.5% ethanol group was probably the most interesting effect seen in this study. A role of anxiety herein was suggested but could not be established with certainty. Other hypotheses were also discussed. The large increase of distance-from-bottom resulting in swimming close to the water surface, which occurred in both buspirone groups and in the 0.5%-ethanol group, is most likely not an anxiolytic response, because of the discrepancy with the in the literature well-established time-course and the absence of any effect of 3.5-hr habituation or black walls on vertical measures. Finally, locomotion and duration freezing could not be specifically taken as indicators for the state of anxiety and the results concerning transparent-wall preference were not sufficient clear. We conclude that the neuronal and ethological mechanisms underlying the effects of habituation, white-aversion, buspirone and ethanol on anxiety-like behavioral responses are complex and need further exploration.

Dissociating the effects of habituation, black walls, buspirone and ethanol on anxiety-like behavioral responses in shoaling zebrafish. A 3D approach to social behavior.

Reversible surface wetting behavior is a hot topic of research because of the potential engineering applications. In the present work, a hierarchical micro/nanostructure is fabricated on brass by alternate current (AC) etching. The superhydrophilic as-prepared etched brass (EB) turns into superhydrophobic after the modification of stearic acid for 1 min. After annealing at 350 °C for 5 min, the superhydrophobic modified EB surface becomes superhydrophilic again. Furthermore, the annealed EB can restore the superhydrophobicity with the remodification of stearic acid. The wetting transition is realized by stearic acid modification and annealing rapidly in 6 min. The wetting transition mechanism is discussed based on the surface chemical analysis. This method is facile and suitable for the construction of large-scale and complex brass surfaces with tunable wetting behaviors.

Rapid reversible superhydrophobicity-to-superhydrophilicity transition on alternating current etched brass.

Wax deposition during the production and transportation of crude oil is a global problem in oil industries. Fabrication of underwater self-cleaning materials can provide a new strategy to prohibit wax deposition. In this paper, conversion films on carbon steel with hierarchical micro/nanostructure are fabricated through a novel in situ alternating-current deposition method. The flower-like conversion films are composed of amorphous iron phosphate and present superhydrophilicity in air and superoleophobicity underwater. The conversion films can efficiently prevent the deposition of wax in water-contained crude oil, showing excellent self-cleaning performance. This facile and low-cost fabrication of a self-cleaning film provides a good strategy for underwater-oil prevention.

Bioinspired in situ growth of conversion films with underwater superoleophobicity and excellent self-cleaning performance.

Because of its highly developed social character, zebrafish is a promising model system for the study of the genetic and neurochemical basis of altered social engagement such as is common in autism and schizophrenia. The traditional shoaling paradigm investigates social cohesion in homogeneous groups of zebrafish. However, the social dynamics of mixed groups is gaining interest from a therapeutic point of view and thus warrants animal modeling. Furthermore, mutant zebrafish are not always available in large numbers. Therefore, we developed a new paradigm that allows exploring shoaling in heterogeneous groups. The effects of MK-801, a non-competitive antagonist of the glutamate N-methyl-D-aspartate (NMDA) receptor, on social cohesion were studied to evaluate the paradigm. The drug has previously been shown to mimic aspects of autism and schizophrenia. Our results show that a single MK-801-treated zebrafish reduced social cohesion of the entire shoal drastically. Preliminary observations suggest that the social dynamics of the shoal as a whole was altered.

Liqun Zhu

R&D

xyZfish

Using an Automated 3D-tracking System to Record Individual and Shoals of Adult Zebrafish

Liqun Zhu

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xyZfish

Using an Automated 3D-tracking System to Record Individual and Shoals of Adult Zebrafish

R&D