Summary of freely-living koalas used at the Koala Conservation Center, Phillip Island, Australia. Heart rate values are given as means ± 1SD, and the range is indicated in parentheses.

<p>Summary of freely-living koalas used at the Koala Conservation Center, Phillip Island, Australia. Heart rate values are given as means ± 1SD, and the range is indicated in parentheses.</p

Heart rates of Crimson (red), Nugget (orange) and Nemo (blue), averaged every 5 minutes (dots) over the whole recording periods.

<p>A curve depicting the moving average of the heart rate, calculated with a 3 points step is superimposed on the dots. Note the drastic change in heart rate around 6:30 PM for all koalas. Arrows indicate the time at which an experimenter walked around the trees of Crimson and Nugget.</p

Heart rates recorded during periods of complete inactivity over a) the day (1:00–6:00 PM) and b) the evening (6:00–10:00 PM) of the experiment for the two koalas in contact with tourists (Nugget and Crimson) and the koala without human contact (Nemo).

<p>Heart rates recorded during periods of complete inactivity over a) the day (1:00–6:00 PM) and b) the evening (6:00–10:00 PM) of the experiment for the two koalas in contact with tourists (Nugget and Crimson) and the koala without human contact (Nemo).</p

Averaged spectral analysis of the heart rate variability over the periods of inactivity of the two koalas in contact with tourists (Nugget and Crimson) and the koala without human contact (Nemo) during a) the day (1:00–6:00 PM) with an enlarged portion highlighting the small LF peaks; and b) the evening (6:00–10:00 PM).

<p>Averaged spectral analysis of the heart rate variability over the periods of inactivity of the two koalas in contact with tourists (Nugget and Crimson) and the koala without human contact (Nemo) during a) the day (1:00–6:00 PM) with an enlarged portion highlighting the small LF peaks; and b) the evening (6:00–10:00 PM).</p

RMSSD and SDNN/RMSSD ratio (see Methods) for a) Nemo, b) Nugget and c) Crimson.

<p>In each figure, the lower and upper curves depict the evolution of the RMSSD and the SDNN/RMSSD, respectively, over the complete (dotted black) recording period. Each period of inactivity (green for RMSSD and blue for the ratio) used to calculate the power spectrum of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007378#pone-0007378-g004" target="_blank">Fig. 4</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007378#pone-0007378-g005" target="_blank">5</a>, are also indicated for information.</p

Spectral analysis of the heart rate variability during periods of inactivity throughout the recording periods for a) Nemo (no contact with human), b) Nugget and c) Crimson.

<p>Spectral analysis of the heart rate variability during periods of inactivity throughout the recording periods for a) Nemo (no contact with human), b) Nugget and c) Crimson.</p

An excerpt of the electrocardiogram signal of Nugget showing the PQRT peaks.

<p>An excerpt of the electrocardiogram signal of Nugget showing the PQRT peaks.</p

TableA1

Bird ID, emergence period, mean and total excursion time, and number of excursions

Immunostaining of chromosomes of <i>Allium</i> species using an anti-AfiCENH3 antibody.

<p>(A), (E), (I) and (M): DAPI stained chromosomes. (B), (F), (J) and (N): Immunosignals of an anti-AfiCENH3 antibody. (C), (G), (K) and (O): Immunosignals of an anti-α-tubulin antibody. (D): Merged image of (A–C). (H): Merged image of (E–G). (L): Merged image of (I–K). (P): Merged image of (M–O). (A–D): <i>A. fistulosum.</i> (E–H): <i>A. cepa.</i> (I–L): <i>A. sativum</i>. (M–P): <i>A. tuberosum.</i> Scale bar, 10 µm.</p

Chromosome Dynamics Visualized with an Anti-Centromeric Histone H3 Antibody in <em>Allium</em>

<div><p>Due to the ease with which chromosomes can be observed, the <em>Allium</em> species, and onion in particular, have been familiar materials employed in cytogenetic experiments in biology. In this study, centromeric histone H3 (CENH3)-coding cDNAs were identified in four <em>Allium</em> species (onion, welsh onion, garlic and garlic chives) and cloned. Anti-CENH3 antibody was then raised against a deduced amino acid sequence of CENH3 of welsh onion. The antibody recognized all CENH3 orthologs of the <em>Allium</em> species tested. Immunostaining with the antibody enabled clear visualization of chromosome behavior during mitosis in the species. Furthermore, three-dimensional (3D) observation of mitotic cell division was achieved by subjecting root sections to immunohistochemical techniques. The 3D dynamics of the cells and position of cell-cycle marker proteins (CENH3 and α-tubulin) were clearly revealed by immunohistochemical staining with the antibodies. The immunohistochemical analysis made it possible to establish an overview of the location of dividing cells in the root tissues. This breakthrough in technique, in addition to the two centromeric DNA sequences isolated from welsh onion by chromatin immuno-precipitation using the antibody, should lead to a better understanding of plant cell division. A phylogenetic analysis of <em>Allium</em> CENH3s together with the previously reported plant CENH3s showed two separate clades for monocot species tested. One clade was made from CENH3s of the <em>Allium</em> species with those of Poaceae species, and the other from CENH3s of a holocentric species (<em>Luzula nivea</em>). These data may imply functional differences of CENH3s between holocentric and monocentric species. Centromeric localization of DNA sequences isolated from welsh onion by chromatin immuno-precipitation (ChIP) using the antibody was confirmed by fluorescence <em>in situ</em> hybridization and ChIP-quantitative PCR.</p> </div