<i>CaP60A</i>^{<i>kum170</i>} mutant has altered heart rate and cardiac dimensions after heat shock.

<p><b>A</b>. Percent paralysis after heat shock. <i>CaP60A</i>^kum170 heterozygote flies were exposed to heat shock of varying durations (no heat shock defined as 0 min (closed circle), 5 min (closed triangles), 7 min (open triangles), or 10 min (open circles)) and observed for up to 72 hours. All flies receiving a heat shock of greater than 7 minutes developed irreversible paralysis; while a smaller percentage of flies developed paralysis after 5-minute heat shock. <b>B</b>. Representative optical coherence tomography (OCT) recordings from <i>w</i>¹¹¹⁸ and <i>CaP60A</i>^kum170 after varying durations of heat shock (HS). End diastolic dimension (EDD) and end systolic dimension (ESD) are denoted in red; A 125 micron standard and one-second bar are shown. <b>C</b>. Heart rates measured from 3 second OCT recordings show a progressive decline in heart rate with increasing durations of heat shock. <b>D</b>. End diastolic dimensions (EDDs) increase with increasing duration of heat shock. <b>E</b>. Fractional shortening is not markedly altered with heat shock in the CaP60A^kum170 mutants. *p<0.05, † p<0.005, ‡p<0.0001 in comparison to 0 minutes heat shock by one-way ANOVA with Tukey’s multiple comparisons test. N= 16, 12, 16, 4, 22 for 0, 5, 6, 7 and 10 minute groups, respectively.</p

Expression of human mutant S151A-SGCD causes an inducible and reversible dilated cardiomyopathy in adult <i>Drosophila</i>.

<p>(A) Temperature shift from 18°C to 26°C causes the induction of S151A-SGCD expression and subsequent deterioration in cardiac function. At 96 hours post induction, flies expressing S151A-SGCD demonstrate an enlargement in EDD and ESD with a resultant impairment in FS. At 96 hours, a temperature shift back to 18°C results in a repression of S151A-SGCD expression and subsequent improvement in cardiac function with return of EDD, ESD and FS to near baseline. A similar level of wt-SGCD expression after temperature shift from 18°C to 26°C does not result in deterioration in cardiac function. Each graph represents the summary data for serial OCT measurements of EDD, ESD and FS and are expressed as the mean +/− SE (n = 16 for wt-SGCD and n = 16 for S151A-SGCD). * P<0.05 for time point measurements compared to baseline and <i>#</i> P<0.05 for measurements between wt-SGCD and S151A-SGCD at each time point. (B) Representative serial OCT m-modes in individual flies expressing S151A-SGCD or wt-SGCD at the indicated times and temperatures. The arrows indicate EDD when the fly heart is fully relaxed and ESD when the fly heart if fully contracted. A 125 micron standard and 1 second scale bar is shown.</p

Expression of S151A-SGCD precedes cardiac function changes in adult <i>Drosophila</i> expressing S151A-SGCD.

<p>(A).Temperature shift from 18°C to 26°C results in deterioration in cardiac function and induction of S151A-SGCD expression. After a temperature shift to 26°C, the induction of transgene expression precedes the development of dilated cardiomyopathy by 48 hours as demonstrated by QRT-PCR measurements of S151A-SGCD mRNA levels compared to cardiac function by serial OCT. A second temperature shift back to 18°C represses S151A-SGCD mRNA expression and results in restoration of cardiac function by 48 hours. We performed three independent experiments using different batches of flies each time. The summary data for QRT-PCR at the indicated times and temperatures are expressed as the mean +/− SE of three independent experiments, each performed in triplicate. * p<0.05 for S151A-SGCD expression at indicted time vs. baseline; # p<0.05 for wt-SGCD expression at the indicated time points vs. baseline; Δ p<0.05 for wt-SGCD vs. S151A-SGCD at the 48 hour time point. (B–D) Summary data for the time course of S151A-SGCD expression (closed circles) vs. cardiac measurements (closed squares) based on serial measurements of EDD, ESD, and FS as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007132#pone-0007132-g001" target="_blank">Figure 1</a>.</p

Rescue of Cardiac function with CaP60A overexpression.

<p><b>A</b>. Representative OCT images of <i>w</i>¹¹¹⁸, <i>heldup</i>² (<i>hdp²</i>) and cardiac specific CaP60A (<i>tinC-wtCaP60A</i>) overexpression in <i>hdp</i>² genetic background (<i>hdp²; tinC-wtCaP60A</i>); Scale bar= 125µm, EDD= End diastolic dimension, ESD=End systolic dimension. <b>B</b>. Heart rate, C. EDD (µm), D. ESD (µm) and <b>E</b>. Fractional shortening (%) in <i>w</i>^<i>1118</i>, <i>CaP60A</i>^kum170 after 10 minute heat shock, <i>tinC>CaP60A </i><i>RNAi</i>, <i>Df</i>(2R) <i>BSC601</i> and <i>CaP60A</i>^{<i>KG00570</i>}. †p<0.001 vs. <i>w</i>¹¹¹⁸, *p<0.05 vs. <i>tinC>CaP60A </i><i>RNAi</i>, ‡p<0.0001 vs. <i>w</i>¹¹¹⁸, **p<0.005 vs. <i>w</i>¹¹¹⁸ by one-way ANOVA with Tukey’s multiple comparisons test.</p

The effects of Calcium Release mutations on the <i>CaP60A</i>^kum170 cardiac phenotypes.

<p><b>A</b>. Heat shock induced paralysis in heterozygous <i>CaP60A</i>^{<i>kum170</i>} flies as well as in transheterozygous mutants carrying both <i>CaP60A</i>^{<i>kum170</i>} and a mutation in Ryanodine receptor (<i>Rya-r44F¹⁶</i>). No heat shock induced paralysis was noted in the single <i>Rya-r44F</i>¹⁶ mutants or <i>w</i>¹¹¹⁸. <b>B</b>. Representative OCT images from <i>w</i>¹¹¹⁸, <i>CaP60A</i>^{<i>kum170</i>}, <i>Rya-r44F</i>¹⁶, <i>CaP60A</i>^{<i>kum170</i>}; <i>Rya-r44F</i>¹⁶ with and without 10-minute heat shock; A 125 micron standard and one-second bar are shown. EDD= End diastolic dimension, ESD=End systolic dimension. <b>C</b>. Heart rate, D. EDD and <b>E</b>. Fractional Shortening (%) in <i>w</i>¹¹¹⁸, <i>CaP60A</i>^{<i>kum170</i>}, <i>Rya-r44F</i>¹⁶, <i>CaP60A</i>^{<i>kum170</i>}<i>; Rya-r44F</i>¹⁶ at baseline and after 10-minute heat shock. *p<0.0002 vs. non-heat shock state of the same genotype, † p<0.0001 vs. <i>CaP60A</i>^{<i>kum170</i>} after heat shock, #p<0.0001 vs. <i>w</i>¹¹¹⁸ of the same treatment condition, ‡p<0.0002 vs. <i>Rya-r44F</i>¹⁶ of the same treatment condition by one-way ANOVA with Tukey’s multiple comparisons test.</p

Cardiac parameters in <i>w¹¹¹⁸, Oregon-R</i>, and the cardiac-specific, inducible driver line.

<p>OCT measurements of end diastolic dimension (EDD) in microns, end systolic dimension (ESD) in microns, and fractional shortening (FS) in percentage are expressed as the mean +/− SE. The values for <i>w¹¹¹⁸</i> (n = 11), <i>Oregon-R</i> (n = 12), the homozygous Gal80^ts; tincΔ4-Gal4 driver line (n = 11) were obtained from flies maintained at 26°C. The values for the Gal80^ts; tincΔ4-Gal4 driver line harboring either wt-human-SGCD (n = 16) or S151A-human-SGCD (n = 16) were obtained from flies maintained at 18°C.</p

The effects of loss of CaP60A function on cardiac parameters.

<p><b>A</b>. Heat shock did not induce paralysis in flies with cardiac specific CaP60A RNAi (<i>tinC>CaP60A RNAi</i>), heterozygous deletion of CaP60A (<i>Df</i>(2R) <i>BSC601</i>), heterozygous p-element disruption of CaP60A (<i>CaP60A^KG00570</i>), or <i>w</i>¹¹¹⁸ compared to <i>CaP60A</i>^{<i>kum170</i>}. <b>B</b>. Representative OCT images in <i>w</i>¹¹¹⁸, <i>CaP60A</i>^kum170 after 10 minute heat shock, <i>tinC>CaP60A </i><i>RNAi</i>, <i>Df</i> (2R) <i>BSC601</i> and <i>CaP60A</i>^{<i>KG00570</i>}; A 125 micron standard and one second bar are shown, C. Heart rate, D. End Diastolic Dimensions (EDD), E. End Systolic Dimensions (ESD) and <b>F</b>. Fractional shortening (%) in <i>w</i>^<i>1118</i>, <i>CaP60A</i>^kum170 after 10 minute heat shock, <i>tinC>CaP60A </i><i>RNAi</i>, <i>Df</i>(2R) <i>BSC601</i> and <i>CaP60A</i>^{<i>KG00570</i>}. †p<0.001 vs. <i>w</i>¹¹¹⁸, *p<0.05 vs. <i>tinC>CaP60A </i><i>RNAi</i>, ‡p<0.0001 vs. <i>w</i>¹¹¹⁸, **p<0.005 vs. <i>w</i>¹¹¹⁸ by one-way ANOVA with Tukey’s multiple comparisons test.</p

Self-Consistent-Charge Density-Functional Tight-Binding (SCC-DFTB) Parameters for Ceria in 0D to 3D

Reducible oxides such as CeO₂ are challenging to describe with standard density-functional theory (DFT) due to the mixed valence states of the cations; they often require the use of non-standard correction schemes, and/or more computationally expensive methods. This adds a new layer of complexity when it comes to the generation of Slater–Koster tables and the corresponding repulsive potentials for self-consistent density-functional based tight-binding (SCC-DFTB) calculations of such materials. In this work, we provide guidelines for how to set up a parametrization scheme for mixed valence oxides within the SCC-DFTB framework, with a focus on reproducing structural and electronic properties as well as redox reaction energies calculated using a reference DFT method. This parametrization procedure was here used to generate parameters for Ce–O systems, with Ce in its +III or +IV formal oxidation states. The generated parameter set is validated by comparison with DFT calculations for various ceria (CeO₂) and reduced ceria (CeO_2–<i>x</i>) systems of different dimensionalities ranging from 0D (nanoparticles) to 3D (bulk). As oxygen vacancy defects in ceria are of crucial importance to many technological applications, special focus is directed toward the capability of describing such defects accurately