Protein acetylation and succinylation are the most crucial protein post-translational modifications (PTMs) involved in the regulation of plant growth and development. In this study, we present the first lysine-acetylation and lysine-succinylation proteome analysis of seedling leaves in Brachypodium distachyon L (Bd). Using high accuracy nano LC-MS/MS combined with affinity purification, we identified a total of 636 lysine-acetylated sites in 353 proteins and 605 lysine-succinylated sites in 262 proteins. These proteins participated in many biology processes, with various molecular functions. In particular, 119 proteins and 115 sites were found to be both acetylated and succinylated, simultaneously. Among the 353 acetylated proteins, 148 had acetylation orthologs in Oryza sativa L., Arabidopsis thaliana, Synechocystis sp. PCC 6803, and Glycine max L. Among the 262 succinylated proteins, 170 of them were found to have homologous proteins in Oryza sativa L., Escherichia coli, Sacchayromyces cerevisiae, or Homo sapiens. Motif-X analysis of the acetylated and succinylated sites identified two new acetylated motifs (K---K and K-I-K) and twelve significantly enriched succinylated motifs for the first time, which could serve as possible binding loci for future studies in plants. Our comprehensive dataset provides a promising starting point for further functional analysis of acetylation and succinylation in Bd and other plant species.
Mounting attention has been focused on defects in macroautophagy/autophagy and the autophagylysosomal pathway (ALP) in cerebral ischemia. TFEB (transcription factor EB)-mediated induction of ALP has been recently considered as the common mechanism in ameliorating the pathological lesion of myocardial ischemia and neurodegenerative diseases. Here we explored the vital role of TFEB in permanent middle cerebral artery occlusion (pMCAO)-mediated dysfunction of ALP and ischemic insult in rats. The results showed that ALP function was first enhanced in the early stage of the ischemic process, especially in neurons of the cortex, and this was accompanied by increased TFEB expression and translocation to the nucleus, which was mediated at least in part through activation by PPP3/ calcineurin. At the later stages of ischemia, a gradual decrease in the level of nuclear TFEB was coupled with a progressive decline in lysosomal activity, accumulation of autophagosomes and autophagy substrates, and exacerbation of the ischemic injury. Notably, neuron-specific overexpression of TFEB significantly enhanced ALP function and rescued the ischemic damage, starting as early as 6 h and even lasting to 48 h after ischemia. Furthermore, neuron-specific knockdown of TFEB markedly reversed the activation of ALP and further aggravated the neurological deficits and ischemic outcome at the early stage of pMCAO. These results highlight neuronal-targeted TFEB as one of the key players in the pMCAO-mediated dysfunction of ALP and ischemic injury, and identify TFEB as a promising target for therapies aimed at neuroprotection in cerebral ischemia.
The pulse is a key biomedical signal containing various human physiological and pathological information highly related to cardiovascular diseases. Pulse signals are often collected from the radial artery based on Traditional Chinese Medicine, or by using flexible pressure sensors. However, the wrist wrapped with a flexible pressure sensor exhibits unstable signals under hand motion because of the concave surface of the wrist. By contrast, fingertips have a convex surface and therefore show great promises in stable and long‐term pulse monitoring. Despite the promising potential, the fingertip pulse signal is weak, calling for highly sensitive detecting devices. Here, a highly sensitive and flexible iontronic pressure sensor with a linear sensitivity of 13.5 kPa−1, a swift response, and remarkable stability over 5000 loading/unloading cycles is developed. This sensor enables stable and high‐resolution detection of pulse waveform under both static condition and finger motion. Fingertip pulse waveforms from subjects of different genders, age, and health conditions are collected and analyzed, suggesting that fingertip pulse information is highly similar to that of the radial artery. This work justifies that fingertip is an ideal platform for pulse signals monitoring, which would be a competitive alternative to existing complex health monitoring systems.
Background: Village doctors, as gatekeepers for the health of rural residents in China, are confronted with adversity in providing the basic public health services (BPHS), which has significantly impeded them from providing high quality BPHS. This study aimed to explore the obstacles and difficulties faced by village doctors in order to improve the quality and efficiency of BPHS provision and increase the health level of the population. Methods: In-depth interviews were employed to conduct this qualitative study. A total of 51 village doctors in four cities of Shandong Province were interviewed. The interviews were transcribed, anonymized, and imported into NVivo11.0 to facilitate management. Thematic framework analysis employing the constant comparison method was applied to the data analysis. Results: The main challenges faced by village doctors comprised the shortage, gender imbalance, and poor education of village doctors; older village doctors in some villages; low income; lack of social security; inappropriate performance assessment; inadequate professional BPHS training; heavy workload; and insufficient cooperation from rural residents, which have exacerbated the quality, efficiency, and accessibility of BPHS to some extent. Conclusions: Village doctors, as the important BPHS providers in rural Shandong, are facing a wide range of challenges. It is urgent for government officials and policy makers to consider these challenges and concentrate on improving the quality of BPHS provision by developing relevant and practical strategies.
The 14-3-3 gene family identified in all eukaryotic organisms is involved in a wide range of biological processes, particularly in resistance to various abiotic stresses. Here, we performed the first comprehensive study on the molecular characterization, phylogenetics, and responses to various abiotic stresses of the 14-3-3 gene family in Brachypodium distachyon L. A total of seven 14-3-3 genes from B. distachyon and 120 from five main lineages among 12 species were identified, which were divided into five well-conserved subfamilies. The molecular structure analysis showed that the plant 14-3-3 gene family is highly evolutionarily conserved, although certain divergence had occurred in different subfamilies. The duplication event investigation revealed that segmental duplication seemed to be the predominant form by which the 14-3-3 gene family had expanded. Moreover, seven critical amino acids were detected, which may contribute to functional divergence. Expression profiling analysis showed that BdGF14 genes were abundantly expressed in the roots, but showed low expression in the meristems. All seven BdGF14 genes showed significant expression changes under various abiotic stresses, including heavy metal, phytohormone, osmotic, and temperature stresses, which might play important roles in responses to multiple abiotic stresses mainly through participating in ABA-dependent signaling and reactive oxygen species-mediated MAPK cascade signaling pathways. In particular, BdGF14 genes generally showed upregulated expression in response to multiple stresses of high temperature, heavy metal, abscisic acid (ABA), and salicylic acid (SA), but downregulated expression under H2O2, NaCl, and polyethylene glycol (PEG) stresses. Meanwhile, dynamic transcriptional expression analysis of BdGF14 genes under longer treatments with heavy metals (Cd2+, Cr3+, Cu2+, and Zn2+) and phytohormone (ABA) and recovery revealed two main expression trends in both roots and leaves: up-down and up-down-up expression from stress treatments to recovery. This study provides new insights into the structures and functions of plant 14-3-3 genes.
Two Chinese bread wheat cultivars, Jinghua 9 and Zhongmai 175, distinct in grain weight and dough quality, were used to study proteome changes in the embryo and endosperm during grain development using a two-dimensional difference gel electrophoresis (2D-DIGE)-based proteomics approach. In total, 138 and 127 differentially expressed protein (DEP) spots representing 116 and 113 unique DEPs were identified in the embryo and endosperm, respectively. Among them, 54 (31%) DEPs were commonly present in both organs while 62 (35%) and 59 (34%) DEPs occurred only in the embryo and endosperm, respectively. Embryonic DEPs are primarily stress-related proteins and involved in carbohydrate and lipid metabolism, while those from the endosperm are related primarily to carbohydrate metabolism and storage. Principal component analysis (PCA) indicated that the proteome differences in the endosperm caused by different cultivars were greater than those by development stages, while the differences in the embryo showed the opposite pattern. Protein-protein interaction (PPI) analysis revealed a complex network centered primarily on enzymes involved in carbohydrate and protein metabolism. The transcriptional levels of fourteen important DEPs encoding genes showed high similarity between organs and cultivars. In particular, some key DEPs of the endosperm, such as phosphoglucomutase, ADP-glucose pyrophosphorylase (AGPase), and sucrose synthase (SUS), showed significantly upregulated expression, indicating their key roles in starch biosynthesis and grain yield. Moreover, upregulated expression of some storage proteins in the endosperm could improve wheat bread-making quality.
In this study, we performed the first integrated physiological and proteomic analysis of the response to drought and recovery from drought, using Brachypodium distachyon L. Roots and leaves. Drought stress resulted in leaves curling, root tips becoming darker in color and significant changes in some physiological parameters. Two-dimensional difference gel electrophoresis (2D-DIGE) identified 78 and 98 differentially accumulated protein (DAP) spots representing 68 and 73 unique proteins responding to drought stress and/or recovery in roots and leaves, respectively. Differences between the root and leaf proteome were most marked for photosynthesis, energy metabolism, and protein metabolism. In particular, some DAPs involved in energy and protein metabolism had contrasting accumulation patterns in roots and leaves. Protein-protein interaction (PPI) analysis of roots and leaves revealed complex protein interaction networks that can generate synergistic responses to drought stress and during recovery from drought. Transcript analysis using quantitative real-time polymerase chain reaction (qRT-PCR) validated the differential expression of key proteins involved in the PPI network. Our integrated physiological and proteomic analysis provides evidence for a synergistic network involved in responses to drought and active during recovery from drought, in Brachypodium roots and leaves.Plants encounter a variety of biotic and abiotic stresses during growth 1 . These stresses unbalance cellular homeostasis and lead to morphological, physiological, and molecular changes 2 . These changes have a negative impact on survival and biomass production, and can reduce final yield by up to 82% 3 . Global warming and climate change may also be exacerbating the effects of abiotic stresses on crop production in many areas of the world today. Previous reports have suggested that a temperature increase of 1 °C can produce a decrease in yield of up to 10% 4 . Drought in particular, severely impairs plant growth and development and limits crop productivity more than any other environmental factor 5,6 . As the climate continues to change, drought may become a more frequent and cause severe problem 7 . Drought stress induces a range of physiological and biochemical responses in plants. Under drought conditions, the plant root cap produces abscisic acid (ABA), which mediates stomatal closure. This in turn, suppresses cell growth, photosynthetic efficiency, and respiration [8][9][10] . Recently, ABA and stomata have been implicated in the regulation of systemic responses to abiotic stresses 11 . In addition, plants generate toxic ions and reactive oxygen species (ROS) that can decrease enzyme activities and damage essential proteins 12 . The generation of ROS occurs early in the response to water-stress, and ROS are key secondary messengers triggering subsequent defensive measures in plants 1,[13][14][15] . Plants have developed a sophisticated and elaborate system for scavenging high levels of ROS using antioxidant enzymes that include superoxid...
The glaciers on Yulong mountain (5596 m a.s.l.), China, are the southernmost in mainland Eurasia. The largest is Baishui glacier No. 1. Up to 90% of the annual precipitation there falls in summer, when warm moisture-rich air masses associated with the southwest monsoon reach the area from the Indian Ocean. The winter climate is influenced by air masses with a continental origin and by the southern branch of the westerlies. The snow that accumulates on Baishui glacier No. 1 includes marine aerosols associated with the summer monsoon, and dust brought in winter from central/west Asia, Africa or the Thar Desert area. Studies in May 2006 at two sites, one in the accumulation area (4900 m) and one in the ablation area (4750 m), revealed differences between the ionic composition of the snow that had accumulated in the 2005/06 winter and that of the snow which had been deposited during the preceding summer monsoon. Differences in the chemistry of the summer-accumulated snow at the two sites probably reflected local differences in ablation and elution rates, rather than differences in ion supplies. Differences in the chemistry of the winter-accumulated snow may reflect the influence of up-valley winds, which bring more crustal material to the lower site.
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