ALBERT

Beschreibung: © The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 13 (2018): e0205015, doi:10.1371/journal.pone.0205015.

Beschreibung: Channelopsins and photo-regulated ion channels make it possible to use light to control electrical activity of cells. This powerful approach has lead to a veritable explosion of applications, though it is limited to changing membrane voltage of the target cells. An enormous potential could be tapped if similar opto-genetic techniques could be extended to the control of chemical signaling pathways. Photopigments from invertebrate photoreceptors are an obvious choice—as they do not bleach upon illumination -however, their functional expression has been problematic. We exploited an unusual opsin, pScop2, recently identified in ciliary photoreceptors of scallop. Phylogenetically, it is closer to vertebrate opsins, and offers the advantage of being a bi-stable photopigment. We inserted its coding sequence and a fluorescent protein reporter into plasmid vectors and demonstrated heterologous expression in various mammalian cell lines. HEK 293 cells were selected as a heterologous system for functional analysis, because wild type cells displayed the largest currents in response to the G-protein activator, GTP-γ-S. A line of HEK cells stably transfected with pScop2 was generated; after reconstitution of the photopigment with retinal, light responses were obtained in some cells, albeit of modest amplitude. In native photoreceptors pScop2 couples to Go; HEK cells express poorly this G-protein, but have a prominent Gq/PLC pathway linked to internal Ca mobilization. To enhance pScop2 competence to tap into this pathway, we swapped its third intracellular loop—important to confer specificity of interaction between 7TMDRs and G-proteins—with that of a Gq-linked opsin which we cloned from microvillar photoreceptors present in the same retina. The chimeric construct was evaluated by a Ca fluorescence assay, and was shown to mediate a robust mobilization of internal calcium in response to illumination. The results project pScop2 as a potentially powerful optogenetic tool to control signaling pathways.

Beschreibung: This work was funded by Colciencias grant FP44842-010-2015 and Connecticut Fund for Science.

Beschreibung: © The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLos One 13 (2018): e0200386, doi:10.1371/journal.pone.0200386.

Beschreibung: Soft robotics is an emerging technology that has shown considerable promise in deep-sea marine biological applications. It is particularly useful in facilitating delicate interactions with fragile marine organisms. This study describes the shipboard design, 3D printing and integration of custom soft robotic manipulators for investigating and interacting with deep-sea organisms. Soft robotics manipulators were tested down to 2224m via a Remotely-Operated Vehicle (ROV) in the Phoenix Islands Protected Area (PIPA) and facilitated the study of a diverse suite of soft-bodied and fragile marine life. Instantaneous feedback from the ROV pilots and biologists allowed for rapid re-design, such as adding “fingernails”, and re-fabrication of soft manipulators at sea. These were then used to successfully grasp fragile deep-sea animals, such as goniasterids and holothurians, which have historically been difficult to collect undamaged via rigid mechanical arms and suction samplers. As scientific expeditions to remote parts of the world are costly and lengthy to plan, on-the-fly soft robot actuator printing offers a real-time solution to better understand and interact with delicate deep-sea environments, soft-bodied, brittle, and otherwise fragile organisms. This also offers a less invasive means of interacting with slow-growing deep marine organisms, some of which can be up to 18,000 years old.

Beschreibung: This work is supported by NOAA OER Grant # NA17OAR0110083 “Exploration of the Seamounts of the Phoenix Islands Protected Area” to RDR, EEC, TMS and DFG and Schmidt Ocean Institute Grant: “What is the Current State of the Deep-Sea Coral Ecosystem in the Phoenix Island Protected Area?” to EEC, RDR, TMS and DFG; NSF Instrument Development for Biological Research Award # 1556164 to RJW and #1556123 to DFG; the National Academies Keck Futures Initiative of the National Academy of Sciences under award #NAKFI DBS21 to RJW and DFG; and NFS Research Fellowship awarded to KPB (#DGE1144152). It is also supported by the Wyss Institute for Biologically Inspired Engineering at Harvard University. We are grateful for the support from the National Geographic Society Innovation Challenge (Grant No.: SP 12-14) to RJW and DFG.

Beschreibung: © The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 13 (2018): e0207532, doi:10.1371/journal.pone.0207532.

Beschreibung: Acoustic standing waves can precisely focus flowing particles or cells into tightly positioned streams for interrogation or downstream separations. The efficiency of an acoustic standing wave device is dependent upon operating at a resonance frequency. Small changes in a system’s temperature and sample salinity can shift the device’s resonance condition, leading to poor focusing. Practical implementation of an acoustic standing wave system requires an automated resonance control system to adjust the standing wave frequency in response to environmental changes. Here we have developed a rigorous approach for quantifying the optimal acoustic focusing frequency at any given environmental condition. We have demonstrated our approach across a wide range of temperature and salinity conditions to provide a robust characterization of how the optimal acoustic focusing resonance frequency shifts across these conditions. To generalize these results, two microfluidic bulk acoustic standing wave systems (a steel capillary and an etched silicon wafer) were examined. Models of these temperature and salinity effects suggest that it is the speed of sound within the liquid sample that dominates the resonance frequency shift. Using these results, a simple reference table can be generated to predict the optimal resonance condition as a function of temperature and salinity. Additionally, we show that there is a local impedance minimum associated with the optimal system resonance. The integration of the environmental results for coarse frequency tuning followed by a local impedance characterization for fine frequency adjustments, yields a highly accurate method of resonance control. Such an approach works across a wide range of environmental conditions, is easy to automate, and could have a significant impact across a wide range of microfluidic acoustic standing wave systems.

Beschreibung: This research was supported by grants from the National Institute of General Medical Sciences of the National Institutes of Health under award number R21GM107805 and the NSF under award number (OCE-1130140 and OCE-1131134) to SWG, RJO, and HMS.

Beschreibung: The work is made available under the Creative Commons CCO public domain dedication.. The definitive version was published in PLoS Biology 16 (2018): e2006333, doi:10.1371/journal.pbio.2006333.

Beschreibung: Our current understanding of biology is heavily based on a small number of genetically tractable model organisms. Most eukaryotic phyla lack such experimental models, and this limits our ability to explore the molecular mechanisms that ultimately define their biology, ecology, and diversity. In particular, marine protists suffer from a paucity of model organisms despite playing critical roles in global nutrient cycles, food webs, and climate. To address this deficit, an initiative was launched in 2015 to foster the development of ecologically and taxonomically diverse marine protist genetic models. The development of new models faces many barriers, some technical and others institutional, and this often discourages the risky, long-term effort that may be required. To lower these barriers and tackle the complexity of this effort, a highly collaborative community-based approach was taken. Herein, we describe this approach, the advances achieved, and the lessons learned by participants in this novel community-based model for research.

Beschreibung: The research efforts, connections, and collaborations described in this paper and protocols.io (https://www.protocols.io/) were supported by the Gordon and Betty Moore Foundation’s Marine Microbiology Initiative.

Beschreibung: © The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 12 (2017): e0188601, doi:10.1371/journal.pone.0188601.

Beschreibung: Many animals go through one or more metamorphoses during their lives, however, the molecular underpinnings of metamorphosis across diverse species are not well understood. Medusozoa (Cnidaria) is a clade of animals with complex life cycles, these life cycles can include a polyp stage that metamorphoses into a medusa (jellyfish). Medusae are produced through a variety of different developmental mechanisms—in some species polyps bud medusae (Hydrozoa), in others medusae are formed through polyp fission (Scyphozoa), while in others medusae are formed through direct transformation of the polyp (Cubozoa). To better understand the molecular mechanisms that may coordinate these different forms of metamorphosis, we tested two compounds first identified to induce metamorphosis in the moon jellyfish Aurelia aurita (indomethacin and 5-methoxy-2-methylindole) on a broad diversity of medusozoan polyps. We discovered that indole-containing compounds trigger metamorphosis across a broad diversity of species. All tested discomedusan polyps metamorphosed in the presence of both compounds, including species representatives of several major lineages within the clade (Pelagiidae, Cyaneidae, both clades of Rhizostomeae). In a cubozoan, low levels of 5-methoxy-2-methylindole reliably induced complete and healthy metamorphosis. In contrast, neither compound induced medusa metamorphosis in a coronate scyphozoan, or medusa production in either hydrozoan tested. Our results support the hypothesis that metamorphosis is mediated by a conserved induction pathway within discomedusan scyphozoans, and possibly cubozoans. However, failure of these compounds to induce metamorphosis in a coronate suggests this induction mechanism may have been lost in this clade, or is convergent between Scyphozoa and Cubozoa.

Beschreibung: National Science Foundation Graduate Research Fellowship (DGE - 1058262; https://www.nsfgrfp.org/general_resources/about) to RRH. Evo-Devo-Eco Network (IOS # 0955517; http://edenrcn.com/) Research Exchange Funds, awarded to RRH. National Science Foundation Rhode Island Established Program to Stimulate Competitive Graduate Research Fellowship to RRH (DEB-1256695; http://web.uri.edu/rinsfepscor/grad-fellowships/). Brown University Ecology and Evolutionary Biology Dissertation Development Grant from the Bushnell Research and Education Fund awarded to RRH.

Beschreibung: © The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 13 (2018): e0191509, doi:10.1371/journal.pone.0191509.

Beschreibung: Wintertime convective mixing plays a pivotal role in the sub-polar North Atlantic spring phytoplankton blooms by favoring phytoplankton survival in the competition between light-dependent production and losses due to grazing and gravitational settling. We use satellite and ocean reanalyses to show that the area-averaged maximum winter mixed layer depth is positively correlated with April chlorophyll concentration in the northern Labrador Sea. A simple theoretical framework is developed to understand the relative roles of winter/spring convection and gravitational sedimentation in spring blooms in this region. Combining climate model simulations that project a weakening of wintertime Labrador Sea convection from Arctic sea ice melt with our framework suggests a potentially significant reduction in the initial fall phytoplankton population that survive the winter to seed the region’s spring bloom by the end of the 21st century.

Beschreibung: KB, LB, PJR and LRL were supported by the Office of Science (BER), U. S. Department of Energy as part of the Regional and Global Climate Modelling (RGCM) Program. SCD acknowledges support from NASA Award NNX15AE65G North Atlantic Aerosol and Marine Ecosystem Study (NAAMES).

Beschreibung: © The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 12 (2017): e0188340, doi:10.1371/journal.pone.0188340.

Beschreibung: Prion diseases include a number of progressive neuropathies involving conformational changes in cellular prion protein (PrPc) that may be fatal sporadic, familial or infectious. Pathological evidence indicated that neurons affected in prion diseases follow a dying-back pattern of degeneration. However, specific cellular processes affected by PrPc that explain such a pattern have not yet been identified. Results from cell biological and pharmacological experiments in isolated squid axoplasm and primary cultured neurons reveal inhibition of fast axonal transport (FAT) as a novel toxic effect elicited by PrPc. Pharmacological, biochemical and cell biological experiments further indicate this toxic effect involves casein kinase 2 (CK2) activation, providing a molecular basis for the toxic effect of PrPc on FAT. CK2 was found to phosphorylate and inhibit light chain subunits of the major motor protein conventional kinesin. Collectively, these findings suggest CK2 as a novel therapeutic target to prevent the gradual loss of neuronal connectivity that characterizes prion diseases.

Beschreibung: This work was supported by Alzheimer Association New Investigator Research Grant to Promote Diversity NIRGD-11-206379 and Consejo Nacional de Investigaciones Científicas y Técnicas PIP 112 20150100954 CO (to GP), National Institutes of Health NS066942A and NS096642 (to GM), R01-NS023868 and R01-NS041170 (to STB).

Beschreibung: © The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 13 (2018): e0190905, doi:10.1371/journal.pone.0190905.

Beschreibung: Trichoplax adhaerens has only six cell types. The function as well as the structure of crystal cells, the least numerous cell type, presented an enigma. Crystal cells are arrayed around the perimeter of the animal and each contains a birefringent crystal. Crystal cells resemble lithocytes in other animals so we looked for evidence they are gravity sensors. Confocal microscopy showed that their cup-shaped nuclei are oriented toward the edge of the animal, and that the crystal shifts downward under the influence of gravity. Some animals spontaneously lack crystal cells and these animals behaved differently upon being tilted vertically than animals with a typical number of crystal cells. EM revealed crystal cell contacts with fiber cells and epithelial cells but these contacts lacked features of synapses. EM spectroscopic analyses showed that crystals consist of the aragonite form of calcium carbonate. We thus provide behavioral evidence that Trichoplax are able to sense gravity, and that crystal cells are likely to be their gravity receptors. Moreover, because placozoans are thought to have evolved during Ediacaran or Cryogenian eras associated with aragonite seas, and their crystals are made of aragonite, they may have acquired gravity sensors during this early era.

Beschreibung: This research was supported by the intramural research program of the NIH, NINDS.

Beschreibung: Author(s): F. Buccella We assume that the two hidden charm pentaquark states discovered at L H C b are built from three light quarks and a c c ¯ pair. Further assumed is that the three light quarks and the c c ¯ pair are both in colour octet states. Thus, for the final J P = 5 2 + state, the three light quarks and the c c ¯ pair are in... [Phys. Rev. D 98, 114011] Published Thu Dec 13, 2018

Beschreibung: Author(s): H. Bahtiyar, K. U. Can, G. Erkol, M. Oka, and T. T. Takahashi We evaluate the spin- 3 / 2 → spin − 1 / 2 electromagnetic transitions of the doubly charmed baryons on 2 + 1 flavor, 3 2 3 × 64 PACS-CS lattices with a pion mass of 156 ( 9 )     MeV / c 2 . A relativistic heavy quark action is employed to minimize the associated systematic errors on charm-quark observables. We extract the... [Phys. Rev. D 98, 114505] Published Thu Dec 13, 2018