Arabidopsis ETHYLENE RESPONSE FACTOR 8 (ERF8) has dual functions in ABA signaling and immunity.

BACKGROUND: ETHYLENE RESPONSE FACTOR (ERF) 8 is a member of one of the largest transcription factor families in plants, the APETALA2/ETHYLENE RESPONSIVE FACTOR (AP2/ERF) superfamily. Members of this superfamily have been implicated in a wide variety of processes such as development and environmental stress responses. RESULTS: In this study we demonstrated that ERF8 is involved in both ABA and immune signaling. ERF8 overexpression induced programmed cell death (PCD) in Arabidopsis and Nicotiana benthamiana. This PCD was salicylic acid (SA)-independent, suggesting that ERF8 acts downstream or independent of SA. ERF8-induced PCD was abolished by mutations within the ERF-associated amphiphilic repression (EAR) motif, indicating ERF8 induces cell death through its transcriptional repression activity. Two immunity-related mitogen-activated protein kinases, MITOGEN-ACTIVATED PROTEIN KINASE 4 (MPK4) and MPK11, were identified as ERF8-interacting proteins and directly phosphorylated ERF8 in vitro. Four putative MPK phosphorylation sites were identified in ERF8, one of which (Ser103) was determined to be the predominantly phosphorylated residue in vitro, while mutation of all four putative phosphorylation sites partially suppressed ERF8-induced cell death in N. benthamiana. Genome-wide transcriptomic analysis and pathogen growth assays confirmed a positive role of ERF8 in mediating immunity, as ERF8 knockdown or overexpression lines conferred compromised or enhanced resistance against the hemibiotrophic bacterial pathogen Pseudomonas syringae, respectively. CONCLUSIONS: Together these data reveal that the ABA-inducible transcriptional repressor ERF8 has dual roles in ABA signaling and pathogen defense, and further highlight the complex influence of ABA on plant-microbe interactions.

A smart fluorescence nanoprobe for the detection of cellular alkaline phosphatase activity and early osteogenic differentiation.

In the past decades, biomaterials were designed to induce stem cell toward osteogenic differentiation. However, conventional methods for evaluation osteogenic differentiation all required a process of cell fixation or lysis, which induce waste of a large number of cells. In this study, a fluorescence nanoprobe was synthesized by combining phosphorylated fluoresceinamine isomer I (FLA) on the surface of mesoporous silica-coated superparamagnetic iron oxide (Fe3O4@mSiO2) nanoparticles. In the presence of alkaline phosphatase (ALP), the phosphorylated FLA on the nanoprobe would be hydrolyzed, resulting in a fluorescence recovery of FLA. During early osteogenic differentiation, a high-level expression of cellular ALP was induced, which accelerated the hydrolysis of phosphorylated FLA, resulting in an enhancement of cellular fluorescence intensity. This fluorescence nanoprobe provides us a rapid and non-toxic method for the detection of cellular ALP activity and early osteogenic differentiation.

Fluorescence light-up AIE probe for monitoring cellular alkaline phosphatase activity and detecting osteogenic differentiation.

Alkaline phosphatase (ALP) is an important monophosphate hydrolase during cell mineralization and osteogenic differentiation. Though traditional methods are provided for evaluating the ALP expression in the fixed and lysed cells, at present it is still challenging to monitor the ALP activity in living cells. In this work, three phosphorylated tetraphenylethylene (TPE) probes (TPE-PA, TPE-2PA and TPE-4PA) with different numbers of -PO3H2 groups were synthesized for monitoring the ALP activity. It was found that in aqueous solution, both the TPE-PA and TPE-2PA probes were highly sensitive to ALP. In the presence of ALP, they could be quickly hydrolysed, resulting in an aggregation-induced emission (AIE) to light up ALP. While in living cells, only TPE-2PA showed good cell penetrability and high fluorescence signal-to-noise ratio during osteogenic differentiation. This probe provides us a new strategy to screen the ALP activity in living stem cells for detecting osteogenic differentiation.

Rational design of multifunctional magnetic mesoporous silica nanoparticle for tumor-targeted magnetic resonance imaging and precise therapy.

In this paper, a multifunctional theranostic magnetic mesoporous silica nanoparticle (MMSN) with magnetic core was developed for magnetic-enhanced tumor-targeted MR imaging and precise therapy. The gatekeeper ß-cyclodextrin (ß-CD) was immobilized on the surface of mesoporous silica shell via platinum(IV) prodrug linking for reduction-triggered intracellular drug release. Then Arg-Gly-Asp (RGD) peptide ligand was further introduced onto the gatekeeper ß-CD via host-guest interaction for cancer targeting purpose. After active-targeting endocytosis by cancer cells, platinum(IV) prodrug in MMSNs would be restored to active platinum(II) drug in response to the innative reducing microenvironment in cancer cells, resulting in the detachment of ß-CD gatekeeper and thus simultaneously triggering the in situ release of anticancer drug doxorubicin (DOX) entrapped in the MMSNs to kill cancer cells. It was found that with the aid of an external magnetic field, drug loaded MMSNs showed high contrast in MR imaging in vivo and exhibited magnetically enhanced accumulation in the cancer site, leading to significant inhibition of cancer growth with minimal side effects. This multifunctional MMSN will find great potential as a theranostic nanoplatform for cancer treatment.

Photo-Activatable Substrates for Site-Specific Differentiation of Stem Cells.

In this report, a UV sensitive, PEGylated PFSSTKTC (Pro-Phe-Ser-Ser-Thr-Lys-Thr-Cys) peptide was modified on quartz substrate to investigate the spatial controlled differentiation of stem cells. This substrate could restrict the cell adhesion due to the steric hindrance of PEG shell. With UV irradiation, PFSSTKTC became exposed owing to the breakage of o-nitrobenzyl group with the detachment of PEG shell. The irradiation boundary on substrate was stable in the long term. The in vitro osteogenic differentiation results revealed that under the site-specific irradiation, the mesenchymal stem cells (MSCs) could specifically differentiate into osteoblast under the induction of PFSSTKTC peptide. This photoactivatable biomaterial shows great potential for region controllable and precise MSCs differentiation.

A novel function of BMHP1 and cBMHP1 peptides to induce the osteogenic differentiation of mesenchymal stem cells.

Bone marrow homing peptide 1 (BMHP1), which was derived from a phage display peptide library (PDPL), is known to be home to bone marrow and bind to stem cells. For the first time, the effect of BMHP1 on the differentiation behavior of mesenchymal stem cells (MSCs) was evaluated. BMHP1 was tethered to modified quartz substrates, and MSCs were seeded on the substrates. It was found that BMHP1 could enhance cell adhesion and proliferation. More importantly, we found that BMHP1 could induce osteogenic differentiation either with a maintenance medium (DMEM) or osteogenic differentiation medium (ODM). Cyclic BMHP1 (cBMHP1) was further synthesized and it was found that cBMHP1 also exhibit a similar, but slightly worse effect on the osteogenic differentiation of MSCs as compared to BMHP1. This work enlightens us on the fact that BMHP1 and cBMHP1 may be used as osteogenic stimulators for MSCs based therapy.

Evaluating the effects of charged oligopeptide motifs coupled with RGD on osteogenic differentiation of mesenchymal stem cells.

Mesenchymal stem cells, due to their multilineage differentiation potential, have emerged as a promising cell candidate for cell-based therapy. In recent years, biomaterials were artificially synthesized to control the differentiation of mesenchymal stem cells. In this study, a series of charged or neutral oligopeptide motifs coupled with RGD were synthesized and used for surface modification using quartz substrates as model. Cell behaviors on the modified surfaces with different charged oligopeptide motifs were studied. It was found that these different charged oligopeptide motifs coupled with RGD were biocompatible for cell proliferation and adhesion. Moreover, it was demonstrated that the positively charged oligopeptide motif could inhibit osteogenic differentiation, while the negatively charged and neutral oligopeptide motifs could enhance osteogenic differentiation in the presence of RGD. This work may bring us enlightenment that different charged oligopeptide motifs coupled with RGD may be used for biomaterial surface modification for different stem cell-based therapies.

The synergistic effect of a BMP-7 derived peptide and cyclic RGD in regulating differentiation behaviours of mesenchymal stem cells.

Precisely controlling the behaviours of stem cells has far-reaching application potential in clinical trials. In this study, we have developed a self-assembled monolayer (SAM) of a cyclic RGD peptide (cycRGD) and bone forming peptide-1 (BFP-1) on a quartz substrate to regulate the behaviours of mesenchymal stem cells (MSCs). The results demonstrated that cycRGD can accelerate the cell adhesion on the substrate, thereby enhancing the ability of BFP-1 in mediating the osteogenic activity. And the synergistic effect between these two functional peptides in osteogenic differentiation of MSCs was confirmed in terms of immunofluorescent staining, Alizarin Red S staining for mineralization and alkaline phosphatase (ALP) activity assay. This finding might give a new insight into designing functional substrates to regulate desired differentiation of stem cells.

Photoresponsive smart template for reversible cell micropatterning.

A novel micropatterned smart template based on transparency photolithography allows the spatial control of cell micropatterning. By utilizing the photoinduced reaction of azobenzene ligands and cyclodextrin-terminated alkanesilane via host-guest recognitions, cells can be easily controlled to adhere reversibly in well defined areas.

The roles of ABA in plant-pathogen interactions.

Defence against abiotic and biotic stresses is crucial for the fitness and survival of plants under adverse or suboptimal growth conditions. The phytohormone abscisic acid (ABA) is not only important for mediating abiotic stress responses, but also plays a multifaceted and pivotal role in plant immunity. This review presents examples demonstrating the importance of crosstalk between ABA and the key biotic stress phytohormone salicylic acid in determining the outcome of plant--pathogen interactions. We then provide an overview of how ABA influences plant defence responses against various phytopathogens with particular emphasis on the Arabidopsis--Pseudomonas syringae model pathosystem. Lastly, we discuss future directions for studies of ABA in plant immunity with emphasis on, its role in the crosstalk between biotic and abiotic stress responses, the importance of distinguishing direct and indirect effects of ABA, as well as the prospect of utilizing the recently elucidated core ABA signaling network to gain further insights into the roles of ABA in plant immunity.