Dendrogram heat map of differential protein levels.

<p>(A) Clustering of proteomics data based on time points. Note that the data corresponding to 9 o’clock contribute major differences. (B) Clustering of proteomics data based on differential proteins.</p

Proteomics analysis reveals a dynamic diurnal pattern of photosynthesis-related pathways in maize leaves

<div><p>Plant leaves exhibit differentiated patterns of photosynthesis rates under diurnal light regulation. Maize leaves show a single-peak pattern without photoinhibition at midday when the light intensity is maximized. This mechanism contributes to highly efficient photosynthesis in maize leaves. To understand the molecular basis of this process, an isobaric tag for relative and absolute quantitation (iTRAQ)-based proteomics analysis was performed to reveal the dynamic pattern of proteins related to photosynthetic reactions. Steady, single-peak and double-peak protein expression patterns were discovered in maize leaves, and antenna proteins in these leaves displayed a steady pattern. In contrast, the photosystem, carbon fixation and citrate pathways were highly controlled by diurnal light intensity. Most enzymes in the limiting steps of these pathways were major sites of regulation. Thus, maize leaves optimize photosynthesis and carbon fixation outside of light harvesting to adapt to the changes in diurnal light intensity at the protein level.</p></div

All-Electrochem-Active Thick Electrode with Dual-Continuous TiO₂‑Carbon Integrated Skeletons for Low-Temperature Lithium Storage

Newly designed all-electrochem-active thick electrode (∼500 μm) with dual-continuous integrated skeletons of defective rutile-anatase TiO2 (D-R-A-TiO2) heterojunctions and carbon have been introduced to enhance efficient electron–ion transport for high-rate energy storage, which provides a new idea for low-temperature lithium storage. For the first time, we anneal anatase TiO2 integrated carbon under CO2 atmosphere for converting anatase to rutile and activating carbon simultaneously, to fabricate freestanding all-electrochem-active thick electrode. The D-R-A-TiO2 heterojunctions contain a type II staggered band alignment, which significantly induce highly localized electrons and lower the migration barrier of ions. The continuous D-R-A-TiO2 heterojunctions form synergistically advantageous electronic networks, and the thick electrode (up to 60.97 mg cm–2) delivers outstanding areal capacity (14.14 mAh cm–2 at 0.61 mA cm–2) under 30 °C. The areal capacity is 8.62 mAh cm–2 at 0.57 mA cm–2 under −10 °C. When the temperature drops to −20 °C, the areal capacity still delivers 4.92 mAh cm–2 at 0.57 mA cm–2. And the D-R-A-TiO2 electrode still delivers 3.2 mAh cm–2 capacity after 70 cycles at 0.57 mA cm–2 under −20 °C

Immunofluorescent staining of cultured NG neurons.

<p>NG neurons were stained using antibodies against P2X1, P2X2, P2X3, and P2X4 receptor subunits (A, D, G, and J, respectively). The nuclei of cultured NG neurons were stained with antibodies against NeuN (B, E, H, K). Merged images (C, F, I, L) representing co-staining of P2X receptor subunits and NeuN are shown. The scale bar shown in L is representative of all images, and represents 50 µm.</p

Four types of I_ATPs and their relationship with NG neuron diameter.

<p>(A) The currents activated by ATP (10⁻⁴ M) in NG were characterized according to fast (F), intermediate (I), slow (S), and very slow (VS) kinetics with an expanded time axis. The horizontal bar above the traces indicates the application of ATP. (B) The 10–90% rising time (ordinate) of different types of ATP (10⁻⁴ M)-activated currents (black circles  =  type F, red triangles  =  type I, green squares  =  for type S, and yellow diamonds  =  type VS I_ATPs) against neuron diameter. Each point in the graph represents a single cell. The correlation coefficient was 0.66 (<i>p</i><0.05). (C) Representative traces of type F, I, S, and VS ATP (10⁻⁴ M)-activated currents. (D) Comparisons of the absolute ratios of the four types of I_ATPs in different subpopulations (small, medium, and large sizes) of NG neurons. Specifically, F type: small (13/54, 28.9%), medium (36/54, 80.0%), and large (5/45, 11.1%); I type: small (5/75, 6.7%), medium (56/75, 74.7%), and large (14/75, 18.6%); S type: small (11/126, 8.7%), medium (94/126, 74.6%), and large (21/126, 16.7%); VS type: small (7/59, 11.9%), medium (34/59, 57.6%), and large (18/59, 30.5%).</p

Expression levels of 15 selected genes that are related to photosynthesis and carbon metabolism.

<p>The expression levels of each gene are presented relative to the first time point (i.e., 6 AM). Values are the means ±standard error (SE) of three independent experiments with at least three replicates each.</p

Generation of iTRAQ proteomics data.

<p>(A) Venn diagram of identified proteins. The numbers indicate the counts of identified maize leaf proteins with differential and steady levels. (B) The distribution of the fold-changes of the identified proteins.</p

Proteins with differential levels during diurnal light cycle that are related to photosynthesis and carbon metabolism.

<p>The class of each protein was k-means clustered based on the dynamic protein levels measured at eight time points.</p

Relevance of the P2X1–4 subunits on the four types of I_ATP.

<p>(A) Schematic view of the setup for the whole cell patch clamp and a representative image of a recorded cell under the phase contrast microscope and immunohistochemistry. (B) Immunohistochemistry revealed positive or negative staining for P2X1–4 subunits, which correlated with the type of I_ATP and cell size. The samples in each row were from four different neurons that responded to ATP with different types of ATP-activated current. P2X3 staining was positive in all four types of I_ATP neurons. P2X1 was positive in F, I, and S I_ATPs, but negative in VS. P2X2 staining was only absent in neurons with type I I_ATP, and P2X4 was positive in neurons with type F, I, and some S I_ATPs.</p

Concentration-response relationships and the efficacy order of P2X receptor antagonists on I_ATPs.

<p>(A) Sequential current traces of F, I, S, and VS ATP-activated currents recorded from rat NG neurons in response to different concentrations of ATP (from 10⁻⁵ to 3×10⁻³ M). Current traces of each type were obtained from the same neuron. (B) The dose-response curves for each type of I_ATPs. Each point represents the means ± SEM of 10–15 neurons. All ATP-induced currents were normalized to the response induced by 3×10⁻³ M ATP in each type. The holding potential was set at −60 mV. The data for ATP were a good fit to the Hill equation I = I_max/[1+ (EC50/C) n], where C is the concentration of ATP, I is the normalized amplitude of I_ATP, and EC50 is the concentration of ATP for the half maximal current response. (C) The efficacy order of the inhibitory effects of P2X receptor antagonists on four distinct I_ATPs. The columns in the bar graph show the inhibitory effects of the P2X receptor antagonists: PPADS (10⁻⁴M), suramin (10⁻⁴ M), and RB2 (10⁻⁴ M). F-type, suramin >PPADS > RB2; I-type, suramin > PPADS > RB2; S-type, suramin > RB2> PPADS; VS-type: suramin > RB2> PPADS. *<i>p</i><0.05, **<i>p</i><0.01.</p