Cumulative survival probability for tumor progression according to resection degree.

<p>Cumulative survival probability for tumor progression according to resection degree.</p

Cumulative survival probability for death according to resection degree.

<p>Cumulative survival probability for death according to resection degree.</p

Summary of hepatobiliary disorders.

<p>Summary of hepatobiliary disorders.</p

Results in a multivariate Cox proportional hazards analysis on PFS.

<p>Results in a multivariate Cox proportional hazards analysis on PFS.</p

Base-line characteristics of the included patients.

<p>Base-line characteristics of the included patients.</p

Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves

<div><p>Although there is a great wealth of data supporting the occurrence of simultaneous synthesis and breakdown of storage carbohydrate in many organisms, previous ¹³CO₂ pulse-chase based studies indicated that starch degradation does not operate in illuminated Arabidopsis leaves. Here we show that leaves of <i>gwd</i>, <i>sex4</i>, <i>bam4</i>, <i>bam1/bam3</i> and <i>amy3/isa3/lda</i> starch breakdown mutants accumulate higher levels of starch than wild type (WT) leaves when cultured under continuous light (CL) conditions. We also show that leaves of CL grown <i>dpe1</i> plants impaired in the plastidic disproportionating enzyme accumulate higher levels of maltotriose than WT leaves, the overall data providing evidence for the occurrence of extensive starch degradation in illuminated leaves. Moreover, we show that leaves of CL grown <i>mex1/pglct</i> plants impaired in the chloroplastic maltose and glucose transporters display a severe dwarf phenotype and accumulate high levels of maltose, strongly indicating that the MEX1 and pGlcT transporters are involved in the export of starch breakdown products to the cytosol to support growth during illumination. To investigate whether starch breakdown products can be recycled back to starch during illumination through a mechanism involving ADP-glucose pyrophosphorylase (AGP) we conducted kinetic analyses of the stable isotope carbon composition (δ¹³C) in starch of leaves of ¹³CO₂ pulsed-chased WT and AGP lacking <i>aps1</i> plants. Notably, the rate of increase of δ¹³C in starch of <i>aps1</i> leaves during the pulse was exceedingly higher than that of WT leaves. Furthermore, δ¹³C decline in starch of <i>aps1</i> leaves during the chase was much faster than that of WT leaves, which provides strong evidence for the occurrence of AGP-mediated cycling of starch breakdown products in illuminated Arabidopsis leaves.</p></div

Leaves of different starch breakdown mutants display a high starch content phenotype when cultured under continuous light conditions.

<p>(A) Iodine staining and (B) starch content in leaves of WT and the indicated starch breakdown mutants cultured under CL conditions. Leaves were harvested at the 18 days after sowing (DAS) growth stage. In “B” values represent the means ± SE determined from three independent experiments using 6 plants in each experiment. Asterisks indicate significant differences with respect to WT plants according to Student´s t-tests (<i>p</i><0.05).</p

Isotope ratio mass spectrometric evidence for the occurrence of starch cycling in illuminated leaves through a mechanism involving AGP.

<p>The graphics represent the values of δ¹³C in starch of leaves of (A) 26 DAS WT (Col-<i>0</i>) and <i>aps1</i> plants, and (B) 22 DAS WT (Ws-2) and <i>pgi1-2</i> plants exposed to ¹³C enriched CO₂ for 5 hours and then chased for 15 additional hours. Plants were cultured in growth cabinets under long day (LD) conditions. The grey area indicates the ¹³CO₂ pulse period. Starch content in leaves is shown in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0171245#pone.0171245.s004" target="_blank">S4 Fig</a></b>. Values represent the means ± SE determined from three independent experiments using 6 plants in each experiment.</p

Amyloglucosidase releases carbon compounds other than starch glucose molecules from Arabidopsis leaf ethanol precipitates.

<p>The graphic shows the total carbon (TOC) and the starch carbon content in amyloglucosidase digests of WT (Col-<i>0</i>) and <i>aps1</i> leaves. Values represent the means ± SE determined from three independent experiments using 6 plants in each experiment.</p

Maltose content in leaves of WT (Col-<i>0</i>), <i>aps1</i>, <i>pgm</i>, <i>aps1/pgm</i>, <i>mex1/aps1</i> and <i>mex1/pgm</i> plants.

<p>Leaves of the indicated plants were harvested at the 18 DAS growth stage. Values obtained using HPAEC-PAD and GC-MS are represented with white and grey columns, respectively. Values represent the means ± SE determined from three independent experiments using 6 plants in each experiment. Asterisks indicate significant differences according to Student´s t-tests (*<i>P</i><0.05, <i>aps1</i>, <i>pgm</i> and <i>aps1/pgm</i> vs. Col-<i>0</i>; **<i>P</i><0.05, <i>mex1/aps1</i> vs. <i>aps1;</i> ***<i>P</i><0.05, <i>mex1/pgm</i> vs. <i>pgm</i>). Values correspond to plants cultured under continuous light (CL) conditions. Essentially the same results were obtained using plants cultured under long day (LD) conditions (not shown).</p