Lack of evidence for direct phosphorylation of recombinantly expressed P2X2 and P2X3 receptors by protein kinase C

P2X3 and P2X2+3 receptors are present on sensory neurons, where they contribute not only to transient nociceptive responses, but also to hypersensitivity underlying pathological pain states elicited by nerve injuries. Increased signalling through P2X3 and P2X2+3 receptors may arise from an increased routing to the plasma membrane and/or gain of function of pre-existing receptors. An obvious effector mechanism for functional modulation is protein kinase C (PKC)-mediated phosphorylation, since all P2X family members share a conserved consensus sequence for PKC, TXR/K, within the intracellularly located N-terminal domain. Contradictory reports have been published regarding the exact role of this motif. In the present study, we confirm that site-directed elimination of the potential phosphor-acceptor threonine or the basic residue in the P+2 position of the TXR/K sequence accelerates desensitization of P2X2 receptors and abolishes P2X3 receptor function. Moreover, the PKC activator phorbol 12-myristate 13-acetate increased P2X3 (but not P2X2) receptor-mediated currents. Biochemically, however, we were unable to demonstrate by various experimental approaches a direct phosphorylation of wild-type P2X2 and P2X3 receptors expressed in both Xenopus laevis oocytes and HEK293 cells. In conclusion, our data support the view that the TXR/K motif plays an important role in P2X function and that phorbol 12-myristate 13-acetate is capable of modulating some P2X receptor subtypes. The underlying mechanism, however, is unlikely to involve direct PKC-mediated P2X receptor phosphorylation

Building a fish passage assessment protocol for New Zealand: implementation of Bayesian network models for estimating fish passage success

Migratory fish species are dependent on connected habitats to complete their life cycles. Instream structures such as culverts, weirs and dams can impede the movement of migratory species. Disruptions to migratory pathways impact ecosystem health by reducing the abundance and diversity of species present. A number of metrics are available for quantifying habitat fragmentation within river networks, but they are dependent on sufficient information being available on the location and severity of migration barriers. Characterising the likelihood of fish passage success at instream structures requires information on the characteristics of the structure and the capabilities of fishes. Biotelemetry and mark-recapture studies are the most effective approaches for quantifying passage success, but are impractical for broad-scale evaluation of multiple instream structures. Bayesian networks offer a flexible approach for deriving probabilistic models suitable for broad-scale rapid assessment of instream structures for barrier severity. We present a Bayesian network derived for evaluating the probability of fish passage success at culverts in New Zealand. A formal expert elicitation process was utilised to populate the prior probability distributions in the model. We present the results from over 350 culverts where the model has been applied. By taking advantage of expert knowledge, the model offers a practical and objective approach for rapidly quantifying the likelihood of fish passage success at multiple instream structures without the need for resource intensive tagging studies. The results are also consistent with requirements for developing environmental reporting metrics for stream connectivity and the model has been used in a new fish passage assessment protocol for New Zealand

Educational Technology Primer: A Guide for Pre-Service Teachers

The Educational Technology Primer grew out of the realization that today’s teacher education students have different needs than their predecessors. Current students arrive on campus less intimidated by technology and more experienced with the use of technology in instruction. However, many introductory educational technology texts still highlight the acquisition of basic technology operations; knowledge and skills that made sense when students entering an introductory educational technology course lacked technology experiences, but not today

Fish passage hydrodynamics New Zealand context

New Zealand is home to 57 native freshwater fish species, of which a considerable number are diadromous, having to move between freshwater and saltwater at least once during their lifecycle. The economic utilisation of New Zealand rivers has largely been carried out without fish migration behaviour in mind, resulting in thousands of structures that prevent fish migration up- and downriver. Remediation of existing structures, especially culverts, and construction of new, more fish friendly, structures requires in-depth knowledge of the needs of the target fish species. In April 2018, the New Zealand Fish Passage Guidelines were released, providing a design framework to enable fish passage in new and existing structures. In support of the new guidance, our project aims to gain insight into the swimming behaviour and performance of inanga (Galaxias maculatus) under various hydraulic conditions, in particular when swimming upstream, which is not well understood. For this purpose, we are designing a new experimental setup in a 600 mm wide flume at the Water Engineering Laboratory at the University of Auckland. We will study hydrodynamics of fish passes with roughened surfaces, baffles and energy dissipators. We will evaluate sensor equipment used to enable flow and fish tracking, with the intention of tracking individual inanga at critical cross sections. This will allow us to study fish response to turbulence, boundary layers, resting zones and wetted margins. We aim to gain valuable insight into design methods and materials that best help inanga, and potentially other members of the family Galaxiidae, with their migration. Eventually, the project aims to provide guidelines suitable for retrofitting existing and building new structures

Practice characteristics of Emergency Department extracorporeal cardiopulmonary resuscitation (eCPR) programs in the United States: The current state of the art of Emergency Department extracorporeal membrane oxygenation (ED ECMO).

PURPOSE: To characterize the current scope and practices of centers performing extracorporeal cardiopulmonary resuscitation (eCPR) on the undifferentiated patient with cardiac arrest in the emergency department. METHODS: We contacted all US centers in January 2016 that had submitted adult eCPR cases to the Extracorporeal Life Support Organization (ELSO) registry and surveyed them, querying for programs that had performed eCPR in the Emergency Department (ED ECMO). Our objective was to characterize the following domains of ED ECMO practice: program characteristics, patient selection, devices and techniques, and personnel. RESULTS: Among 99 centers queried, 70 responded. Among these, 36 centers performed ED ECMO. Nearly 93% of programs are based at academic/teaching hospitals. 65% of programs are less than 5 years old, and 60% of programs perform ≤3 cases per year. Most programs (90%) had inpatient eCPR or salvage ECMO programs prior to starting ED ECMO programs. The majority of programs do not have formal inclusion and exclusion criteria. Most programs preferentially obtain vascular access via the percutaneous route (70%) and many (40%) use mechanical CPR during cannulation. The most commonly used console is the Maquet Rotaflow(®). Cannulation is most often performed by cardiothoracic (CT) surgery, and nearly all programs (\u3e85%) involve CT surgeons, perfusionists, and pharmacists. CONCLUSIONS: Over a third of centers that submitted adult eCPR cases to ELSO have performed ED ECMO. These programs are largely based at academic hospitals, new, and have low volumes. They do not have many formal inclusion or exclusion criteria, and devices and techniques are variable

Propuesta de un modelo de Oficina de Gestión de Proyectos I+D+i en universidades públicas manabitas

Public universities have as their main design the satisfaction of social needs. This is achieved through the implementation of research, development and innovation (R+D+i) projects that comprise the systematic work undertaken in order to increase the knowledge base and devise new utilities. However, the complexity of this type of project and the competitiveness of the current environment warrants the need to systematize management activities. Hence the importance of creating University Project Management Offices (PMOs), because in addition to being entities that support project management, they contribute to monitoring and control in terms of costs, deadlines, and quality. The objective of the research was to propose a PMO model that contributes to improving the management of R+D+i projects in manabitas public universities. A diagnosis of project management maturity in universities was carried out, which showed an intermediate level of maturity. Additionally, through interviews applied to the vice-chancellors for research, it was verified that in none of the universities there were specific entities that functioned as PMOs, nor was a consolidated project management culture actually appreciated. The proposal of the PMO model that guarantees the success of these projects.Las universidades públicas tienen como principal designio la satisfacción de las necesidades de sociales. Esto se logra a través de la implementación de proyectos de investigación, desarrollo e innovación(I+D+i), que comprenden el trabajo sistemático emprendido con la finalidad de acrecentar el acervo de conocimientos e idear nuevas utilidades. Sin embargo, la complejidad de esta tipología de proyectos y la competitividad del entorno actual amerita la necesidad de sistematizar las actividades de gestión. De ahí la importancia de crear Oficinas de Gestión de Proyectos (PMO) universitarias, porque además de ser entidades que apoyan la gestión de los proyectos, contribuyen a llevar un monitoreo y control en términos de costos, plazos y calidad. La investigación tuvo como objetivo proponer un modelo de PMO que contribuya a mejorar la dirección de proyectos I+D+i en las universidades públicas manabitas. Fue realizado un diagnóstico de la madurez de gestión de proyectos en las universidades, lo cual evidenció un nivel intermedio de madurez. Adicionalmente, a través de entrevistas aplicadas a los vicerrectores de investigación se comprobó que en ninguna de las universidades existían entidades específicas que funcionaran como PMO, ni se apreciaba en realidad cultura de gestión de proyectos consolidada. Con relación a los resultados derivados fue realizada la propuesta del modelo de PMO que garantice el éxito de estos proyectos

Not just a migration problem: Metapopulations, habitat shifts and gene flow are also important for fishway science and management

Worldwide, fishways are increasingly criticized for failing to meet conservation goals. We argue that this is largely due to the dominance of diadromous species of the Northern Hemisphere (e.g., Salmonidae) in the research that underpins the concepts and methods of fishway science and management. With highly diverse life histories, swimming abilities and spatial ecologies, most freshwater fish species do not conform to the stereotype imposed by this framework. This is leading to a global proliferation of fishways that are often unsuitable for native species. The vast majority of fish populations do not undertake extensive migrations between clearly separated critical habitats, yet the movement of individuals and the genetic information they carry is critically important for population viability. We briefly review some of the latest advances in spatial ecological modelling for dendritic networks to better define what it means to achieve effective fish passage at a barrier. Through a combination of critical habitat assessment and the modelling of metapopulations, climate change�driven habitat shifts, and adaptive gene flow, we recommend a conceptual and methodological framework for fishway target�setting and monitoring suitable for a wide range of species. In the process, we raise a number of issues that should contribute to the ongoing debate about fish passage research and the design and monitoring of fishways

A Location-based File Sharing Application

Multiple computing devices in close proximity must usually rely on the Internet in order to share information, even though doing so is grossly inefficient and subject to external factors. A method to facilitate this sort of local sharing in a secure manner could help alleviate these issues. This study proposes to demonstrate the substantial demand for a more efficient and interactive means to exchange information among networks of people. We will detail how this project will result in a software protocol capable of linking mobile devices for the purpose of sending and receiving data through manipulation of available technology, pursuit of developing computer systems, and creation of an innovative program. The tasks at hand do not rely on innovation through "brute force" development of new hardware, but rather on manipulation of existing technology through revolutionary software

Photoreversible interconversion of a phytochrome photosensory module in the crystalline state.

A major barrier to defining the structural intermediates that arise during the reversible photointerconversion of phytochromes between their biologically inactive and active states has been the lack of crystals that faithfully undergo this transition within the crystal lattice. Here, we describe a crystalline form of the cyclic GMP phosphodiesterases/adenylyl cyclase/FhlA (GAF) domain from the cyanobacteriochrome PixJ in Thermosynechococcus elongatus assembled with phycocyanobilin that permits reversible photoconversion between the blue light-absorbing Pb and green light-absorbing Pg states, as well as thermal reversion of Pg back to Pb. The X-ray crystallographic structure of Pb matches previous models, including autocatalytic conversion of phycocyanobilin to phycoviolobilin upon binding and its tandem thioether linkage to the GAF domain. Cryocrystallography at 150 K, which compared diffraction data from a single crystal as Pb or after irradiation with blue light, detected photoconversion product(s) based on Fobs - Fobs difference maps that were consistent with rotation of the bonds connecting pyrrole rings C and D. Further spectroscopic analyses showed that phycoviolobilin is susceptible to X-ray radiation damage, especially as Pg, during single-crystal X-ray diffraction analyses, which could complicate fine mapping of the various intermediate states. Fortunately, we found that PixJ crystals are amenable to serial femtosecond crystallography (SFX) analyses using X-ray free-electron lasers (XFELs). As proof of principle, we solved by room temperature SFX the GAF domain structure of Pb to 1.55-Å resolution, which was strongly congruent with synchrotron-based models. Analysis of these crystals by SFX should now enable structural characterization of the early events that drive phytochrome photoconversion

A location-based communication platform: integrating file sharing with interpersonal contact

Gemstone Team FLIP (File Lending in Proximity)Sharing on the Internet, even among computing devices in close proximity, is both inefficient and inconvenient. Online services and websites do not take advantage of easily obtainable geo-locational data that can improve sharing. We at Team FLIP have extended an existing mapping system called TerpNav with functionality that allows proximate users to interact and collaborate while sharing digital information. This study demonstrates both the feasibility of and demand for a more efficient and interactive method to exchange information among proximate networks of people