Gust forecasting by using numerical weather prediction and on-site measurement

This paper was reviewed and accepted by the APCWE-IX Programme Committee for Presentation at the 9th Asia-Pacific Conference on Wind Engineering, University of Auckland, Auckland, New Zealand, held from 3-7 December 2017

Gust forecasting by using numerical weather prediction and on-site measurement

This paper was reviewed and accepted by the APCWE-IX Programme Committee for Presentation at the 9th Asia-Pacific Conference on Wind Engineering, University of Auckland, Auckland, New Zealand, held from 3-7 December 2017

A Prediction Model for Lateral Force Coefficients of a Train Car in Turbulent Flows

This paper was reviewed and accepted by the APCWE-IX Programme Committee for Presentation at the 9th Asia-Pacific Conference on Wind Engineering, University of Auckland, Auckland, New Zealand, held from 3-7 December 2017

An improved inverse-type Ca²⁺ indicator can detect putative neuronal inhibition in <i>Caenorhabditis elegans</i> by increasing signal intensity upon Ca²⁺ decrease

<div><p>Sensory processing is regulated by the coordinated excitation and inhibition of neurons in neuronal circuits. The analysis of neuronal activities has greatly benefited from the recent development of genetically encoded Ca²⁺ indicators (GECIs). These molecules change their fluorescence intensities or colours in response to changing levels of Ca²⁺ and can, therefore, be used to sensitively monitor intracellular Ca²⁺ concentration, which enables the detection of neuronal excitation, including action potentials. These GECIs were developed to monitor increases in Ca²⁺ concentration; therefore, neuronal inhibition cannot be sensitively detected by these GECIs. To overcome this difficulty, we hypothesised that an inverse-type of GECI, whose fluorescence intensity increases as Ca²⁺ levels decrease, could sensitively monitor reducing intracellular Ca²⁺ concentrations. We, therefore, developed a Ca²⁺ indicator named inverse-pericam 2.0 (IP2.0) whose fluorescent intensity decreases 25-fold upon Ca²⁺ binding <i>in vitro</i>. Using IP2.0, we successfully detected putative neuronal inhibition by monitoring the decrease in intracellular Ca²⁺ concentration in AWC^ON and ASEL neurons in <i>Caenorhabditis elegans</i>. Therefore, IP2.0 is a useful tool for studying neuronal inhibition and for the detailed analysis of neuronal activities <i>in vivo</i>.</p></div

<i>In vitro</i> properties of IP2.0.

<p><b>A</b>, Normalized fluorescence excitation (dashed lines) and emission (solid lines) spectra in Ca²⁺-free (green and blue lines) and Ca²⁺-saturated (orange and red lines) states. <b>B</b>, pH-dependency of normalized amplitudes at the 515 nm excitation peak in Ca²⁺-free (blue line) and Ca²⁺-saturated (red line) states. <b>C</b>, Ca²⁺ titration curve of inverse-pericam (left) and IP2.0 (right). <b>D</b>, Fluorescence rise response of inverse-pericam (green line) and IP2.0 (blue line) to a stepped decrease in [Ca²⁺]_free from 10 μM to < 10 nM. The raw data of Fig 2D is available in figshare (<a href="https://doi.org/10.6084/m9.figshare.5976067.v1" target="_blank">https://doi.org/10.6084/m9.figshare.5976067.v1</a>).</p

Schematic structures and sequences of inverse-pericam and IP2.0.

<p>Sequences of linkers and amino and substitutions are shown below and above the bars, respectively. His-6: the polyhistidine tag.</p

<i>In vivo</i> imaging of Ca²⁺ responses in <i>C</i>.<i>elegans</i>.

<p>(A) GCaMP6f Ca²⁺ response to isoamylalcohol in AWC^ON (n = 13). (B) IP2.0 Ca²⁺ response to isoamylalcohol in AWC^ON (n = 12). (C) Dual-colour of RCaMP2.0 and IP2.0 Ca²⁺ responses to isoamylalcohol in AWC^ON (n = 10). (D) IP2.0 Ca²⁺ response to change of NaCl concentration in ASEL (n = 8). The values are shown as relative to <i>F</i>_<i>0</i> and error bars represent SEM. The raw data of Fig 4 is available in figshare (<a href="https://doi.org/10.6084/m9.figshare.5976619.v1" target="_blank">https://doi.org/10.6084/m9.figshare.5976619.v1</a>).</p

Representative Ca²⁺ imaging in HeLa cells.

<p>Fluorescence images of HeLa cells (<b>A, C</b>) and fluorescence intensity vs. time traces (<b>B, D</b>) in the ROIs of fluorescence images. Images were taken of HeLa cells transfected with inverse-pericam (<b>A, B</b>) and IP2.0 (<b>C, D</b>). Scale bar: 20 μm. The raw data of Fig 3B and D is available in figshare (<a href="https://doi.org/10.6084/m9.figshare.5976610.v1" target="_blank">https://doi.org/10.6084/m9.figshare.5976610.v1</a>).</p

Spectral characteristics of inverse-pericam and IP2.0.

<p>Spectral characteristics of inverse-pericam and IP2.0.</p

Nuclei detected by the proposed method.

<p>True positives, false positives, and false negatives are shown as red, cyan, and yellow ellipses, respectively. Original image is the same as <a href="http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1004970#pcbi.1004970.g001" target="_blank">Fig 1A</a>.</p