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Background: Cell therapy can protect cardiomyocytes from hypoxia, primarily via paracrine secretions, including extracellular vesicles (EVs). Since EVs fulfil specific biological functions based on their cellular origin, we hypothesised that EVs from human cardiac stromal cells (CMSCLCs) obtained from coronary artery bypass surgery may have cardioprotective properties. Objectives: This study characterises CMSCLC EVs (C_EVs), miRNA cargo, cardioprotective efficacy and transcriptomic modulation of hypoxic human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). C_EVs are compared to bone marrow mesenchymal stromal cell EVs (B_EVs) which are a known therapeutic EV type. Methods: Cells were characterised for surface markers, gene expression and differentiation potential. EVs were compared for yield, phenotype, and ability to protect hiPSC-CMs from hypoxia/reoxygenation injury. EV dose was normalised by both protein concentration and particle count, allowing direct comparison. C_EV and B_EV miRNA cargo was profiled and RNA-seq was performed on EV-treated hypoxic hiPSC-CMs, then data were integrated by multi-omics. Confirmatory experiments were carried out using miRNA mimics. Results: At the same dose, C_EVs were more effective than B_EVs at protecting CM integrity, reducing apoptotic markers, and cell death during hypoxia. While C_EVs and B_EVs shared 70–77% similarity in miRNA content, C_EVs contained unique miRNAs, including miR-202-5p, miR-451a and miR-142-3p. Delivering miRNA mimics confirmed that miR-1260a and miR-202/451a/142 were cardioprotective, and the latter upregulated protective pathways similar to whole C_EVs. Conclusions: This study demonstrates the potential of cardiac tissues, routinely discarded following surgery, as a valuable source of EVs for myocardial infarction therapy. We also identify miR-1260a as protective of CM hypoxia. [ABSTRACT FROM AUTHOR]

Accurate and timely identification of infections is crucial for effective treatment and controlling disease spread. While single biomarkers have traditionally served diagnostic purposes, their sensitivity and specificity limitations call for a more comprehensive approach. Simultaneous detection of multiple biomolecules shows promise in improving accuracy, expediting analysis, and enhancing pathology screening. Therefore, there is a pressing need to develop a noninvasive biosensor capable of quantifying various infection-associated biomarkers concurrently, such as albumin proteins and bacterial molecules. In this endeavor, a novel biosensor has been developed, leveraging a color-indicating optical platform that utilizes gold nanoisland films (AuNIFs) enveloped by cholesteric liquid crystals (CLCs). The integration of CLCs and AuNIFs capitalizes on their sensitive interfacial interactions, highlighting the potential for robust biosensing. Changes in biomolecule concentrations induce discernible alterations in the CLCs–AuNIFs interface alignment, profoundly impacting the hybrid plasmonic–photonic behavior of AuNIFs. The resulting CLCs–AuNIFs biosensor demonstrates exceptional sensitivity and precision in detecting E. coli concentrations and albumin levels. Notably, the biosensor achieves a label-free limit of detection, with an impressive sensitivity of 1 × 106 CFU/ml for E. coli and 10 ng/ml for bovine serum albumin (BSA). This innovative biosensor offers rapid detection, visual clarity, label-free operation, and the ability to detect multiple concentrations of both microbial agents and albumin. Changes in biomolecule concentrations induce noticeable shifts in interface alignment, thereby modulating the hybrid plasmonic–photonic dynamics of AuNIFs. The potential applications of this advancement span biomedical, microbial, and industrial sectors. This unified biosensor has the potential to revolutionize infection-related diagnostics and containment measures, serving as an ideal point-of-care solution. This ground-breaking development leads the way in multi-biomarker detection in infection diagnosis, with profound implications for future of infection diagnostics and therapeutic interventions. [ABSTRACT FROM AUTHOR]

Purpose: This study aims to investigate the impact of regular Kung Fu and Taekwondo practice on the health and quality of life among elderly individuals in the Maule region, Chile. Design/methodology/approach: The authors designed a 12-week Kung Fu and Taekwondo workshop with activities suitable for their age. Through semistructured interviews (at the beginning and the end of the workshop), along with periodic monitoring of vital signs and cardiovascular components, the authors observed an improvement in participants' physical (strength, speed of reaction and flexibility) and psychological conditions (self-esteem and resilience), quality of life (relationships with family and friends and ability to deal with stressful events in working life) and health (waist circumference, percentage of oxygen saturation in blood, blood pressure, among other values). Findings: From these results, the authors affirm that this workshop improves health and physical condition and helps the participants develop the coping capacity to deal with stressful situations and complicated interpersonal relationships. In this sense, the authors conclude that Kung Fu and Taekwondo as regular sports activities can benefit senior citizens' aging process. Originality/value: This research is based on an original study project. [ABSTRACT FROM AUTHOR]

Nanomaterials are widely used in various fields, and ongoing research is focused on developing safe and sustainable nanomaterials. Using zebrafish as a model organism for studying the potentially toxic effects of nanomaterials highlights the importance of developing safe and sustainable nanomaterials. Studies conducted on nanomaterials and their toxicity and potential risks to human and environmental health are vital in biomedical sciences. In the present review, we discuss the potential toxicity of nanomaterials (inorganic and organic) and exposure risks based on size, shape, and concentration. The review further explores various types of nanomaterials and their impacts on zebrafish at different levels, indicating that exposure to nanomaterials can lead to developmental defects, changes in gene expressions, and various toxicities. The review also covers the importance of considering natural organic matter and chorion membranes in standardized nanotoxicity testing. While some nanomaterials are biologically compatible, metal and semiconductor nanomaterials that enter the water environment can increase toxicity to aquatic creatures and can potentially accumulate in the human body. Further investigations are necessary to assess the safety of nanomaterials and their impacts on the environment and human health. [ABSTRACT FROM AUTHOR]

We have achieved a successful synthesis of a label-free, flexible, portable, and colour-indicating biosensor using cholesteric nanostructure liquid crystals (LCs), commonly referred to as cholesteric LCs (CLCs), on paper fibres. This innovative approach holds significant promise for rapid screening in precision medicine and other applications. We incorporated optically active nematic LCs in various configurations within CLCs, allowing them to rotate along different axes. When exposed to light, these LCs reflected light at specific wavelengths. By employing polarised optical microscopy and a charged couple device (CCD), we conducted titration experiments with bovine serum albumin (BSA) on the paper substrate and quantified protein concentrations based on the observed colour shifts. Utilising internal Python pixel analysis software, we evaluated the red, green, and blue colour components, which served as biomarkers for different BSA concentrations. Our results demonstrated a linear correlation between BSA concentration and colour variations. This CLC-based paper-substrate biosensor exhibits the potential to efficiently screen a large number of samples, thereby enhancing the diagnostic accuracy in clinical settings. Combining the colour-indicating and quantifiable attributes of CLCs with the affordability and portability of paper substrates, we have developed a versatile biosensor. Its ease of use, rapid response, and high sensitivity make it a promising tool for future disease detection endeavours. [ABSTRACT FROM AUTHOR]

BiVO4 with suitable band edges and a small band gap is considered an efficient photocatalyst for the oxygen evolution reaction (OER), but short charge-transfer lengths limit its photocatalytic ability. Increasing active sites can essentially enhance oxidation reactions and photocatalytic ability. The alkaline etching technique can refine the surface properties of BiVO4 by introducing oxygen vacancies and possibly establishing a heterojunction, but suitable alkaline etching methods are rarely discussed. In this study, three alkaline etching methods, including hydrothermal, soaking, and electrodeposition processes, are first applied for a constant duration to tailor the surface properties of BiVO4 as a photocatalyst for the OER. Alkaline-etched BiVO4 prepared using hydrothermal and soaking processes shows better photocatalytic abilities due to the generation of additional oxygen vacancies, while poor photocatalytic ability is found for that fabricated using electrodeposition due to the formation of less photocatalytic VO2. The highest photocurrent density of 2.38 mA cm−2 is obtained at 1.23 V versus a reversible hydrogen electrode in an electrolyte without a hole scavenger for the BiVO4 etched in the hydrothermal process, while the untreated BiVO4 presents a photocurrent density of 0.58 mA cm−2. A photocurrent retention of 92.3% is achieved after 5000 seconds of continuous illumination. The study supplies blueprints for establishing effective alkaline etching methods for enhancing the photocatalytic ability of BiVO4, with the exception of time costs. [ABSTRACT FROM AUTHOR]

The diversification of effector function, driven by a co-evolutionary arms race, enables pathogens to establish compatible interactions with hosts. Structurally conserved plant pathogenesis-related PR-1 and PR-1-like (PR-1L) proteins are involved in plant defense and fungal virulence, respectively. It is unclear how fungal PR-1L counters plant defense. Here, we show that Ustilago maydis UmPR-1La and yeast ScPRY1, with conserved phenolic resistance functions, are Ser/Thr-rich region mediated cell-surface localization proteins. However, UmPR-1La has gained specialized activity in sensing phenolics and eliciting hyphal-like formation to guide fungal growth in plants. Additionally, U. maydis hijacks maize cathepsin B-like 3 (CatB3) to release functional CAPE-like peptides by cleaving UmPR-1La's conserved CNYD motif, subverting plant CAPE-primed immunity and promoting fungal virulence. Surprisingly, CatB3 avoids cleavage of plant PR-1s, despite the presence of the same conserved CNYD motif. Our work highlights that UmPR-1La has acquired additional dual roles to suppress plant defense and sustain the infection process of fungal pathogens. Plant PR-1 proteins participate in defense responses against pathogens. Here, the authors show that PR-1-like proteins from the plant pathogenic fungus Ustilago maydis are important for virulence by detecting plant-derived phenolics and modulating plant PR-1-mediated defenses. [ABSTRACT FROM AUTHOR]

This study conducted a systematic review following the guidelines of the PRISMA statement, with the aim of analyzing the scientific production on Education for Sustainable Development (ESD) in in-service and pre-service mathematics teachers. Out of 211 articles identified in the indexed databases, WOS, SCOPUS, and SciELO, 16 were selected for an exhaustive analysis. It was determined that this topic is still in the process of development, focusing on aspects such as learning content, pedagogy, and the learning environment. Most studies employed qualitative methodologies. Contextualization in sustainability is the most advanced competency. The results highlight the importance of continuing to investigate and develop effective strategies to integrate ESD in the continuous education and training of mathematics teachers. It is essential to address the development of competencies in ecological and mediational facets, ensure solid knowledge of mathematical content, employ adequate pedagogical strategies, and understand the relationship between the environment and mathematics. These findings offer insights into the prevailing trend of integrating ESD into mathematics education through ESD elements and didactic competencies, as evidenced by recent studies involving both practicing and pre-service teachers. This understanding may promote the enhancement of the quality of mathematics education practices within this domain. [ABSTRACT FROM AUTHOR]

Effective phosphoproteome of nanoscale sample analysis remains a daunting task, primarily due to significant sample loss associated with non-specific surface adsorption during enrichment of low stoichiometric phosphopeptide. We develop a tandem tip phosphoproteomics sample preparation method that is capable of sample cleanup and enrichment without additional sample transfer, and its integration with our recently developed SOP (Surfactant-assisted One-Pot sample preparation) and iBASIL (improved Boosting to Amplify Signal with Isobaric Labeling) approaches provides a streamlined workflow enabling sensitive, high-throughput nanoscale phosphoproteome measurements. This approach significantly reduces both sample loss and processing time, allowing the identification of >3000 (>9500) phosphopeptides from 1 (10) µg of cell lysate using the label-free method without a spectral library. It also enables precise quantification of ~600 phosphopeptides from 100 sorted cells (single-cell level input for the enriched phosphopeptides) and ~700 phosphopeptides from human spleen tissue voxels with a spatial resolution of 200 µm (equivalent to ~100 cells) in a high-throughput manner. The new workflow opens avenues for phosphoproteome profiling of mass-limited samples at the low nanogram level. A streamlined tandem tip-based workflow for sensitive nanoscale phosphoproteomics is developed, reducing sample loss and processing time, allowing the phosphoproteome profiling of mass-limited samples at the low nanogram level. [ABSTRACT FROM AUTHOR]

Abiotic stresses have significant impacts on crop yield and quality. Even though significant efforts during the past decade have been devoted to uncovering the core signaling pathways associated with the phytohormone abscisic acid (ABA) and abiotic stress in plants, abiotic stress signaling mechanisms in most crops remain largely unclear. The core components of the ABA signaling pathway, including early events in the osmotic stress-induced phosphorylation network, have recently been elucidated in Arabidopsis with the aid of phosphoproteomics technologies. We now know that SNF1-related kinases 2 (SnRK2s) are not only inhibited by the clade A type 2C protein phosphatases (PP2Cs) through dephosphorylation, but also phosphorylated and activated by upstream mitogen-activated protein kinase kinase kinases (MAP3Ks). Through describing the course of studies to elucidate abiotic stress and ABA signaling, we will discuss how we can take advantage of the latest innovations in mass-spectrometry-based phosphoproteomics and structural proteomics to boost our investigation of plant regulation and responses to ABA and abiotic stress. [ABSTRACT FROM AUTHOR]

Myocardial infarction is a major cause of morbidity and mortality worldwide. Due to poor inherent regeneration of the adult mammalian myocardium and challenges with effective drug delivery, there has been little progress in regenerative therapies. Nanocarriers, including liposomes, nanoparticles, and exosomes, offer many potential advantages for the therapy of myocardial infarction, including improved delivery, retention, and prolonged activity of therapeutics. However, there are many challenges that have prevented the widespread clinical use of these technologies. This review aims to summarize significant principles and developments in the field, with a focus on nanocarriers using ligand-based or cell mimicry-based targeting. Lastly, a discussion of limitations and potential future direction is provided. [ABSTRACT FROM AUTHOR]

Steady-state visual evoked potential (SSVEP) has been used to implement brain-computer interface (BCI) due to its advantages of high information transfer rate (ITR) and high accuracy. In recent years, owing to the developments of head-mounted device (HMD), the HMD has become a popular device to implement SSVEP–based BCI. However, an HMD with fixed frame rate only can flash at its subharmonic frequencies which limits the available number of stimulation frequencies for SSVEP-based BCI. In order to increase the number of available commands for SSVEP-based BCI, we proposed a phase-approaching (PA) method to generate visual stimulation sequences at user-specified frequency on an HMD. The flickering sequence generated by our PA method (PAS sequence) tries to approximate user-specified stimulation frequency by means of minimizing the difference of accumulated phases between our PAS sequence and the ideal wave of user-specified frequency. The generated sequence of PA method determines the brightness state for each frame to approach the accumulated phase of the ideal wave. The SSVEPs evoked from stimulators, driven by PAS sequences, were analyzed using canonical correlation analysis (CCA) to identify user’s gazed target. In this study, a six-command SSVEP-based BCI was designed to operate a flying drone. The ITR and detection accuracy are 36.84 bits/min and 93.30%, respectively. [ABSTRACT FROM AUTHOR]

MED25 has been implicated as a negative regulator of the abscisic acid (ABA) signaling pathway. However, it is unclear whether other Mediator subunits could associate with MED25 to participate in the ABA response. Here, we used affinity purification followed by mass spectrometry to uncover Mediator subunits that associate with MED25 in transgenic plants. We found that at least 26 Mediator subunits, belonging to the head, middle, tail, and CDK8 kinase modules, were co‐purified with MED25 in vivo. Interestingly, the tail module subunit MED16 was identified to associate with MED25 under both mock and ABA treatments. We further showed that the disruption of MED16 led to reduced ABA sensitivity compared to the wild type. Transcriptomic analysis revealed that the expression of several ABA‐responsive genes was significantly lower in med16 than those in wild type. Furthermore, we discovered that MED16 may possibly compete with MED25 to interact with the key transcription factor ABA INSENSITIVE 5 (ABI5) to positively regulate ABA signaling. Consistently, med16 and med25 mutants displayed opposite phenotypes in ABA response, cuticle permeability, and differential ABI5‐mediated EM1 and EM6 expression. Together, our data indicate that MED16 and MED25 differentially regulate ABA signaling by antagonistically affecting ABI5‐mediated transcription in Arabidopsis. [ABSTRACT FROM AUTHOR]

Large numbers of cells are generally required for quantitative global proteome profiling due to surface adsorption losses associated with sample processing. Such bulk measurement obscures important cell-to-cell variability (cell heterogeneity) and makes proteomic profiling impossible for rare cell populations (e.g., circulating tumor cells (CTCs)). Here we report a surfactant-assisted one-pot sample preparation coupled with mass spectrometry (MS) method termed SOP-MS for label-free global single-cell proteomics. SOP-MS capitalizes on the combination of a MS-compatible nonionic surfactant, n-Dodecyl-β-D-maltoside, and hydrophobic surface-based low-bind tubes or multi-well plates for 'all-in-one' one-pot sample preparation. This 'all-in-one' method including elimination of all sample transfer steps maximally reduces surface adsorption losses for effective processing of single cells, thus improving detection sensitivity for single-cell proteomics. This method allows convenient label-free quantification of hundreds of proteins from single human cells and ~1200 proteins from small tissue sections (close to ~20 cells). When applied to a patient CTC-derived xenograft (PCDX) model at the single-cell resolution, SOP-MS can reveal distinct protein signatures between primary tumor cells and early metastatic lung cells, which are related to the selection pressure of anti-tumor immunity during breast cancer metastasis. The approach paves the way for routine, precise, quantitative single-cell proteomics. Tsai, Zhang, Scholten et al. develop a surfactant- assisted one-pot sample preparation coupled with mass spectrometry method (SOP-MS) for label-free global single-cell proteomics. This method allows researchers to measure hundreds of proteins from single human cells, suggesting its utility for quantitative single-cell proteomics. [ABSTRACT FROM AUTHOR]

Summary: CDK8 is a key subunit of Mediator complex, a large multiprotein complex that is a fundamental part of the conserved eukaryotic transcriptional machinery. However, the biological functions of CDK8 in plant abiotic stress responses remain largely unexplored.Here, we demonstrated CDK8 as a critical regulator in the abscisic acid (ABA) signaling and drought response pathways in Arabidopsis. Compared to wild‐type, cdk8 mutants showed reduced sensitivity to ABA, impaired stomatal apertures and hypersensitivity to drought stress. Transcriptomic and chromatin immunoprecipitation analysis revealed that CDK8 positively regulates the transcription of several ABA‐responsive genes, probably through promoting the recruitment of RNA polymerase II to their promoters.We discovered that both CDK8 and SnRK2.6 interact physically with an ERF/AP2 transcription factor RAP2.6, which can directly bind to the promoters of RD29A and COLD‐REGULATED 15A (COR15A) with GCC or DRE elements, thereby promoting their expression. Importantly, we also showed that CDK8 is essential for the ABA‐induced expression of RAP2.6 and RAP2.6‐mediated upregulation of ABA‐responsive genes, indicating that CDK8 could link the SnRK2.6‐mediated ABA signaling to RNA polymerase II to promote immediate transcriptional response to ABA and drought signals.Overall, our data provide new insights into the roles of CDK8 in modulating ABA signaling and drought responses. [ABSTRACT FROM AUTHOR]

Summary: In Arabidopsis, the plasma membrane transporter PUT3 is important to maintain the cellular homeostasis of polyamines and plays a role in stabilizing mRNAs of some heat‐inducible genes. The plasma membrane Na+/H+ transporter SOS1 and the protein kinase SOS2 are two salt‐tolerance determinants crucial for maintaining intracellular Na+ and K+ homeostasis.Here, we report that PUT3 genetically and physically interacts with SOS1 and SOS2, and these interactions modulate PUT3 transport activity.Overexpression of PUT3 (PUT3OE) results in hypersensitivity of the transgenic plants to polyamine and paraquat. The hypersensitivity of PUT3OE is inhibited by the sos1 and sos2 mutations, which indicates that SOS1 and SOS2 are required for PUT3 transport activity. A protein interaction assay revealed that PUT3 physically interacts with SOS1 and SOS2 in yeast and plant cells. SOS2 phosphorylates PUT3 both in vitro and in vivo. SOS1 and SOS2 synergistically activate the polyamine transport activity of PUT3, and PUT3 also modulates SOS1 activity by activating SOS2 in yeast cells.Overall, our findings suggest that both plasma‐membrane proteins PUT3 and SOS1 could form a complex with the protein kinase SOS2 in response to stress conditions and modulate the transport activity of each other through protein interactions and phosphorylation. [ABSTRACT FROM AUTHOR]

Osmoregulation is important for plant growth, development and response to environmental changes. SNF1-related protein kinase 2s (SnRK2s) are quickly activated by osmotic stress and are central components in osmotic stress and abscisic acid (ABA) signaling pathways; however, the upstream components required for SnRK2 activation and early osmotic stress signaling are still unknown. Here, we report a critical role for B2, B3 and B4 subfamilies of Raf-like kinases (RAFs) in early osmotic stress as well as ABA signaling in Arabidopsis thaliana. B2, B3 and B4 RAFs are quickly activated by osmotic stress and are required for phosphorylation and activation of SnRK2s. Analyses of high-order mutants of RAFs reveal critical roles of the RAFs in osmotic stress tolerance and ABA responses as well as in growth and development. Our findings uncover a kinase cascade mediating osmoregulation in higher plants. Rapid activation of SnRK2 kinases is central to plant responses to osmotic stress and abscisic acid. Here the authors show that a group of Raf-like kinases are very quickly activated by osmotic stress, and then phosphorylate and activate SnRK2s. [ABSTRACT FROM AUTHOR]

Summary: The capability to maintain cell wall integrity is critical for plants to adapt to unfavourable conditions. l‐Arabinose (Ara) is a constituent of several cell wall polysaccharides and many cell wall‐localised glycoproteins, but so far the contribution of Ara metabolism to abiotic stress tolerance is still poorly understood.Here, we report that mutations in the MUR4 (also known as HSR8) gene, which is required for the biosynthesis of UDP‐Arap in Arabidopsis, led to reduced root elongation under high concentrations of NaCl, KCl, NaNO3, or KNO3.The short root phenotype of the mur4/hsr8 mutants under high salinity is rescued by exogenous Ara or gum arabic, a commercial product of arabinogalactan proteins (AGPs) from Acacia senegal. Mutation of the MUR4 gene led to abnormal cell−cell adhesion under salt stress. MUR4 forms either a homodimer or heterodimers with its isoforms. Analysis of the higher order mutants of MUR4 with its three paralogues, MURL,DUR,MEE25, reveals that the paralogues of MUR4 also contribute to the biosynthesis of UDP‐Ara and are critical for root elongation.Taken together, our work revealed the importance of the Ara metabolism in salt stress tolerance and also provides new insights into the enzymes involved in the UDP‐Ara biosynthesis in plants. [ABSTRACT FROM AUTHOR]

Environmental-friendly nanocomposite hydrogels of carboxymethyl-hexanoyl chitosan (CHC), graphene oxide (GO) and cellulose nanocrystals (CNCs) were combined to produce a bio-hydrogel with great biocompatibility and antibacterial ability. The size of the GO nanosheets was about 200–500 nm, and the CNCs had a length of 100–200 nm and a width of 10–20 nm, as shown by transmission electron microscopy (TEM). X-ray photoelectron spectroscopy (XPS) was utilized for the analysis of the oxygen functional groups of GO. The homogeneous dispersion of the CHC/GO/CNC nanocomposite hydrogel showed significantly higher water absorption capacity and water retention capability. In addition, inhibition of a variety of microorganisms (gram-negative and gram-positive bacteria and fungi) by the introduction of the CHC/GO/CNC nanocomposite hydrogel demonstrated that there is a great opportunity to use it in the bio-medical field, such as for plastic masks and wound dressings. [ABSTRACT FROM AUTHOR]

Respiration is a crucial vital sign to indicate acidotic state of human body. In this paper, we have developed a Holo-Hilbert spectral analysis (HHSA)-based approach to detect subject’s respiration frequency from wrist photoplethysmogram (PPG) signals. The HHSA is a two-layer EMD architecture which first decomposes the original signal into intrinsic mode functions and then tries to find additive and multiplicative interactions among the participating components. The HHSA enables frequency-modulated (FM) and amplitude-modulated (AM) features of a signal can be comprehensively presented on a Holo-Hilbert spectrum (HHS). With the help of HHSA, subject’s respiration frequency can be identified on HHS by finding the AM frequency with the most prominent amplitude around the FM frequency of heart rate. The efficacy of the proposed method has been demonstrated in two designed experiments. In the first experiment, 75 subjects with ages ranged from 20–80 years old, and the difference between the detected results of proposed method and the readouts of transthoracic impedance plethysmography is 0.04 ± 0.96 breathes-per-minute (brm). In the second experiment, six subjects were requested to breathe at 6, 12, 18, 24 brm, following the pacing rhythms of a metronome. The difference between detected respiration rates and expected breathing rates is 0.01 ± 0.70 brm. The HHSA-based approach has manifested its capability to extract respiration-induced multiplicative component in PPG signal. It avoids the ambiguity of frequency representation in signal multiplication when using traditional additive decomposition methods. [ABSTRACT FROM AUTHOR]

Several nucleoporins in the nuclear pore complex (NPC) have been reported to be involved in abiotic stress responses in plants. However, the molecular mechanism of how NPC regulates abiotic stress responses, especially the expression of stress responsive genes remains poorly understood. From a forward genetics screen using an abiotic stress-responsive luciferase reporter (RD29A-LUC) in the sickle-1 (sic-1) mutant background, we identified a suppressor caused by a mutation in NUCLEOPORIN 85 (NUP85), which exhibited reduced expression of RD29A-LUC in response to ABA and salt stress. Consistently, the ABA and salinity induced expression of several stress responsive genes such as RD29A, COR15A and COR47 was significantly compromised in nup85 mutants and other nucleoporin mutants such as nup160 and hos1. Subsequently, Immunoprecipitation and mass spectrometry analysis revealed that NUP85 is potentially associated with HOS1 and other nucleoporins within the nup107-160 complex, along with several mediator subunits. We further showed that there is a direct physical interaction between MED18 and NUP85. Similar to NUP85 mutations, MED18 mutation was also found to attenuate expression of stress responsive genes. Taken together, we not only revealed the involvement of NUP85 and other nucleoporins in regulating ABA and salt stress responses, but also uncovered a potential relation between NPC and mediator complex in modulating the gene expression in plants. [ABSTRACT FROM AUTHOR]

Phosphorylation has an incredible impact on the biological behavior of proteins, altering everything from intrinsic activity to cellular localization and complex formation. It is no surprise then that this post-translational modification has been the subject of intense study and that, with the advent of faster, more accurate instrumentation, the number of large-scale mass spectrometry-based phosphoproteomic studies has swelled over the past decade. Recent developments in sample preparation, phosphorylation enrichment, quantification, and data analysis strategies permit both targeted and ultra-deep phosphoproteome profiling, but challenges remain in pinpointing biologically relevant phosphorylation events. We describe here technological advances that have facilitated phosphoproteomic analysis of cells, tissues, and biofluids and note applications to neuropathologies in which the phosphorylation machinery may be dysregulated, much as it is in cancer. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) regulate gene expression and play critical roles in growth and development as well as stress responses in eukaryotes. miRNA biogenesis in plants requires a processing complex that consists of the core components DICER-LIKE 1 (DCL1), SERRATE (SE) and HYPONASTIC LEAVES (HYL1). Here we show that inactivation of functionally redundant members of the SnRK2 kinases, which are the core components of abscisic acid (ABA) and osmotic stress signaling pathways, leads to reduction in miRNA accumulation under stress conditions. Further analysis revealed that the steady state level of HYL1 protein in plants under osmotic stress is dependent on the SnRK2 kinases. Additionally, our results suggest that the SnRK2 kinases physically associate with the miRNA processing components SE and HYL1 and can phosphorylate these proteins in vitro. These findings reveal an important role for the SnRK2 kinases in the regulation of miRNA accumulation and establish a mechanism by which ABA and osmotic stress signaling is linked to miRNA biogenesis. [ABSTRACT FROM AUTHOR]

This paper studies the amplitude-frequency characteristic of frontal steady-state visual evoked potential (SSVEP) and its feasibility as a control signal for brain computer interface (BCI). SSVEPs induced by different stimulation frequencies, from 13 ~ 31 Hz in 2 Hz steps, were measured in eight young subjects, eight elders and seven ALS patients. Each subject was requested to participate in a calibration study and an application study. The calibration study was designed to find the amplitude-frequency characteristics of SSVEPs recorded from Oz and Fpz positions, while the application study was designed to test the feasibility of using frontal SSVEP to control a two-command SSVEP-based BCI. The SSVEP amplitude was detected by an epoch-average process which enables artifact-contaminated epochs can be removed. The seven ALS patients were severely impaired, and four patients, who were incapable of completing our BCI task, were excluded from calculation of BCI performance. The averaged accuracies, command transfer intervals and information transfer rates in operating frontal SSVEP-based BCI were 96.1%, 3.43 s/command, and 14.42 bits/min in young subjects; 91.8%, 6.22 s/command, and 6.16 bits/min in elders; 81.2%, 12.14 s/command, and 1.51 bits/min in ALS patients, respectively. The frontal SSVEP could be an alternative choice to design SSVEP-based BCI. [ABSTRACT FROM PUBLISHER]

DNA demethylation mediated by the DNA glycosylase ROS1 helps determine genomic DNA methylation patterns and protects active genes from being silenced. However, little is known about the mechanism of regulation of ROS1 enzymatic activity. Using a forward genetic screen, we identified an anti-silencing (ASI) factor, ASI3, the dysfunction of which causes transgene promoter hyper-methylation and silencing. Map-based cloning identified ASI3 as MET18, a component of the cytosolic iron-sulfur cluster assembly (CIA) pathway. Mutation in MET18 leads to hyper-methylation at thousands of genomic loci, the majority of which overlap with hypermethylated loci identified in ros1 and ros1dml2dml3 mutants. Affinity purification followed by mass spectrometry indicated that ROS1 physically associates with MET18 and other CIA components. Yeast two-hybrid and split luciferase assays showed that ROS1 can directly interact with MET18 and another CIA component, AE7. Site-directed mutagenesis of ROS1 indicated that the conserved iron-sulfur motif is indispensable for ROS1 enzymatic activity. Our results suggest that ROS1-mediated active DNA demethylation requires MET18-dependent transfer of the iron-sulfur cluster, highlighting an important role of the CIA pathway in epigenetic regulation. [ABSTRACT FROM AUTHOR]

Measurement of protein phosphorylation plays an essential role in delineating cell signaling pathways. Although the detection of a specific phosphoprotein has been largely accomplished by immunological methods, a specific and sensitive assay to measure total protein phosphorylation level in complex samples such as whole cell extracts has yet to be established. Here, we present a sensitive phosphorylation assay on a microwell plate to determine total protein phosphorylation level calibrated to a phosphoprotein standard. The core of the assay is a reagent termed pIMAGO that is multi-functionalized with titanium ions for its superior selectivity towards phosphorylated proteins and with fluorophores for quantification. The specificity, sensitivity, and quantitative nature of the assay were demonstrated with standard proteins and whole cell lysates. The method was then employed to measure the overall protein phosphorylation level of human cells under different treatments. At last, we investigated the practicability of the assay to serve as a sensitive tool to estimate the amount of phosphorylated samples prior to a mass spectrometry-based phosphoproteomic analysis. [ABSTRACT FROM AUTHOR]

Cathepsin B is one of the major lysosomal cysteine proteases that plays an important role in apoptosis. Herein, we investigated whether Cathepsin B is involved in cardiomyocyte apoptosis caused by hyperthermic injury (HI) and heat shock protein (HSP)-70 protects these cells from HI-induced apoptosis mediated by Cathepsin. HI was produced in H9C2 cells by putting them in a circulating 43 °C water bath for 120 min, whereas preinduction of HSP-70 was produced in H9C2 cells by mild heat preconditioning (or putting them in 42 °C water bath for 30 min) 8 h before the start of HI. It was found that HI caused both cardiomyocyte apoptosis and increased Cathepsin B activity in H9C2 cells. E-64-c, in addition to reducing Cathepsin B activity, significantly attenuated HI-induced cardiomyocyte apoptosis (evidenced by increased apoptotic cell numbers, increased tuncated Bid (t-Bid), increased cytochrome C, increased caspase-9/-3, and decreased Bcl-2/Bax) in H9C2 cells. In addition, preinduction of HSP-70 by mild heat preconditioning or inhibition of HSP-70 by Tripolide significantly attenuated or exacerbated respectively both the cardiomyocyte apoptosis and increased Cathepsin B activity in H9C2 cells. Furthermore, the beneficial effects of pre-induction of HSP-70 by mild heat production in reducing both cardiomyocyte apoptosis and increased Cathepsin B activity caused by HI can be significantly reduced by Triptolide preconditioning. These results indicate that Cathepsin B is involved in HI-induced cardiomyocyte apoptosis in H9C2 cells and HSP-70 protects these cells from HI-induced cardiomyocyte apoptosis through Cathepsin B pathways. [ABSTRACT FROM AUTHOR]

Intolerance to heat exposure is believed to be associated with hypothalamo-pituitary-adrenocortical (HPA) axis impairment [reflected by decreases in blood concentrations of both adrenocorticotrophic-hormone (ACTH) and corticosterone]. The purpose of this study was to determine the effect of human recombinant factor VIIa (rfVIIa) on heat intolerance, HPA axis impairment, and hypothalamic inflammation, ischemic and oxidative damage, and apoptosis in mice under heat stress. Immediately after heat stress (41.2 °C for 1 h), mice were treated with vehicle (1 mL/kg of body weight) or rfVIIa (65-270 µg/kg of body weight) and then returned to room temperature (26 °C). Mice still alive on day 4 of heat exposure were considered survivors. Cellular ischemia markers (e.g., glutamate, lactate-to-pyruvate ratio), oxidative damage markers (e.g., nitric oxide metabolite, hydroxyl radials), and pro-inflammatory cytokines (e.g., interleukin-6, interleukin-1β, tumor necrosis factor-α) in hypothalamus were determined. In addition, blood concentrations of both ACTH and corticosterone were measured. Hypothalamic cell damage was assessed by determing the neuronal damage scores, whereas the hypothalamic cell apoptosis was determined by assessing the numbers of cells stained with terminal deoxynucleotidyl transferase-mediated αUTP nick-end labeling, caspase-3-positive cells, and platelet endothelial cell adhesion molecula-1-positive cells in hypothalamus. Compared with vehicle-treated heated mice, rfVIIa-treated heated mice had significantly higher fractional survival (8/10 vs 1/10), lesser thermoregulatory deficit (34.1 vs 24.8 °C), lesser extents of ischemic, oxidative, and inflammatory markers in hypothalamus, lesser neuronal damage scores and apoptosis in hypothalamus, and lesser HPA axis impairment. Human recombinant factor VIIa appears to exert a protective effect against heatstroke by attenuating hypothalamic cell apoptosis (due to ischemic, inflammatory, and oxidative damage) in mice. [ABSTRACT FROM AUTHOR]

RATIONALE Despite advances in mass spectrometry (MS)-based identification, effective phosphopeptide enrichment is a prerequisite towards comprehensive phosphoproteomic analysis. Based on the different binding affinities and coordination geometries of the Ti4+ and Fe3+ ions with the phosphate group, we report a complementary metal-directed immobilized metal ion affinity chromatography (IMAC) method to increase the identification coverage of a phosphoproteome. METHODS Phosphopeptides from standard phosphoproteins and Raji B cells were enriched from Ti4+-IMAC and Fe3+-IMAC methods, followed by matrix-assisted laser desorption/ionization (MALDI) MS and Orbitrap MS analysis. Optimal enrichment specificity was achieved by selection of acid structure/concentration and organic solvent to compete with non-phosphopeptides. The effect of the metal ion and the chelating compound was evaluated by the comparison of the characteristics of enriched phosphopeptides between Ti4+-IMAC, Fe3+-IMAC and TiO2 methods. RESULTS To address the low enrichment specificity of the Ti4+-IMAC method, a simple one-step acid/solvent controlled IMAC method was developed with significantly improved specificity (88%) and recovery (93%). The most striking discovery is that the optimal Ti4+-IMAC and Fe3+-IMAC methods have low overlapping percentage (10%) among the 2905 enriched phosphopeptides from Raji cells, comprised of the distinct characteristics including hydrophobicity, amino acid compositions, and frequency of multiple phosphorylation of the phosphopeptides. CONCLUSIONS The reported Fe3+-IMAC and Ti4+-IMAC methods can complementarily enrich acidic and basic phosphopeptides to effectively increase the identification coverage of an heterogeneous phosphoproteome (twice than the single approach). Given the reproducibility and low sample loss, the combination of our enrichment strategy with a quantitative technique could be feasible for quantitative phosphoproteomics. Copyright © 2012 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor. Patients diagnosed with GBM have a poor prognosis, and it has been reported that tumor malignancy and GBM recurrence are promoted by STAT3 signaling. As resveratrol (RV), a polyphenol in grapes, is reported to be a potent and non-toxic cancer-preventive compound, the aim of this study was to investigate the therapeutic effect and molecular mechanisms of RV on GBM-derived radioresistant tumor initiating cells (TIC). Firstly, our results showed that primary GBM-CD133+ TIC presented high tumorigenic and radiochemoresistant properties as well as increased protein levels of phosphorylated STAT3. We consistently observed that treatment with shRNA-STAT3 (sh-STAT3) or AG490, a STAT3 inhibitor, significantly inhibited the cancer stem-like cell properties and radioresistance of GBM-CD133+ in vitro and in vivo. Furthermore, treatment of GBM-CD133+ with 100 µM RV induced apoptosis and enhanced radiosensitivity by suppressing STAT3 signaling. Microarray results suggested that RV or AG490 inhibited the stemness gene signatures of GBM-CD133+ and facilitated the differentiation of GBM-CD133+ into GBM-CD133− or astrocytoma cells. Finally, xenotransplant experiments indicated that RV or sh-STAT3 therapy could significantly improve the survival rate and synergistically enhance the radiosensitivity of radiation-treated GBM-TIC. In summary, RV can reduce in vivo tumorigenicity and enhance the sensitivity of GBM-TIC to radiotherapies through the STAT3 pathway. J. Cell. Physiol. 227: 976-993, 2012. © 2011 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Introduction: CD133 (PROM1) is a potential marker for cancer stem cells (CSCs), including those found in brain tumors. Recently, medulloblastoma (MB)-derived CD133-positive cells were found to have CSC-like properties and were proposed to be important contributors to tumorigenicity, cancer progression, and chemoradioresistance. However, the biomolecular pathways and therapeutic targets specific to MB-derived CSCs remain unresolved. Materials and methods: In the present study, we isolated CD133 cells from MB cell lines and determined that they showed increased tumorigenicity, radioresistance, and higher expression of both embryonic stem cell-related and drug resistance-related genes compared to CD133 cells. Bioinformatics analysis suggested that the STAT3 pathway might be important in MB and CD133 cells. To evaluate the effects of inhibiting the STAT3 pathway, MB-derived CD133 cells were treated with the potent STAT3 inhibitor, cucurbitacin I. Treatment with cucurbitacin I significantly suppressed the CSC-like properties and stemness gene signature of MB-derived CD133 cells. Furthermore, cucurbitacin I treatment increased the apoptotic sensitivity of MB-derived CD133 cells to radiation and chemotherapeutic drugs. Notably, cucurbitacin I demonstrated synergistic effects with ionizing radiation to inhibit tumorigenicity in MB-CD133-inoculated mice. Results: These results indicate that the STAT3 pathway plays a key role in mediating CSC properties in MB-derived CD133 cells. Targeting STAT3 with cucurbitacin I may therefore represent a novel therapeutic approach for treating malignant brain tumors. [ABSTRACT FROM AUTHOR]

Soybean (Glycine max L.) is a major crop providing important source for protein and oil for human life. Low phosphate (LP) availability is a critical limiting factor affecting soybean production. Soybean plants develop a series of strategies to adapt to phosphate (Pi) limitation condition. However, the underlying molecular mechanisms responsible for LP stress response remain largely unknown. Here, we performed a label-free quantification (LFQ) analysis of soybean leaves grown under low and high phosphate conditions. We identified 267 induced and 440 reduced differential proteins from phosphate-starved leaves. Almost a quarter of the LP decreased proteins are involved in translation processes, while the LP increased proteins are accumulated in chlorophyll biosynthetic and carbon metabolic processes. Among these induced proteins, an enolase protein, GmENO2a was found to be mostly induced protein. On the transcriptional level, GmENO2a and GmENO2b, but not GmENO2c or GmENO2d, were dramatically induced by phosphate starvation. Among 14 enolase genes, only GmENO2a and GmENO2b genes contain the P1BS motif in their promoter regions. Furthermore, GmENO2b was specifically induced in the GmPHR31 overexpressing soybean plants. Our findings provide molecular insights into how soybean plants tune basic carbon metabolic pathway to adapt to Pi deprivation through the ENO2 enzymes. [ABSTRACT FROM AUTHOR]

We have successfully fabricated poly(3,4-ethylenedioxythiophene) (PEDOT) derivative nanohybrid coatings on flexible SUS316L stainless steel by electrochemical polymerization, which can offer anti-fouling and anti-bacterial capabilities. PEDOT derivative nanohybrids were prepared from polystyrene sulfonates (PSS) and graphene oxide (GO) incorporated into a conducting polymer of PEDOT. Additionally, the negative charge of the PEDOT/GO substrate was further modified by poly-diallyldimethylammonium chloride (PDDA) to form a positively charged surface. These PEDOT derivative nanohybrid coatings could provide a straightforward means of controlling the surface energy, roughness, and charges with the addition of various derivatives in the electrochemical polymerization and electrostatically absorbed process. The characteristics of the PEDOT derivative nanohybrid coatings were evaluated by Raman spectroscopy, scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), water contact angle, and surface potential (zeta potential). The results show that PEDOT/PSS and PEDOT/GO nanohybrid coatings exhibit excellent anti-fouling capability. Only 0.1% of bacteria can be adhered on the surface due to the lower surface roughness and negative charge surface by PEDOT/PSS and PEDOT/GO modification. Furthermore, the anti-bacterial capability (7 mm of inhibition zone) was observed after adding PDDA on the PEDOT/GO substrates, suggesting that the positive charge of the PEDOT/GO/PDDA substrate can effectively kill bacteria (Staphylococcus aureus). Given their anti-fouling and anti-bacterial capabilities, PEDOT derivative nanohybrid coatings have the potential to be applied to biomedical devices such as cardiovascular stents and surgical apparatus. [ABSTRACT FROM AUTHOR]

Magnetic reduced graphene oxide (MRGO) sheets were prepared by embedding Fe3O4 nanoparticles on polyvinylpyrrolidone (PVP) and poly(diallyldimethylammonium chloride) (PDDA)-modified graphene oxide (GO) sheets for bacteria capture and destruction under a high-frequency magnetic field (HFMF). The characteristics of MRGO sheets were evaluated systematically by transmission electron microscopy (TEM), scanning electron microscopy (SEM), zeta potential measurement, X-ray diffraction (XRD), vibrating sample magnetometry (VSM), and X-ray photoelectron spectroscopy (XPS). TEM observation revealed that magnetic nanoparticles (8–10 nm) were dispersed on MRGO sheets. VSM measurements confirmed the superparamagnetic characteristics of the MRGO sheets. Under HFMF exposure, the temperature of MRGO sheets increased from 25 to 42 °C. Furthermore, we investigated the capability of MRGO sheets to capture and destroy bacteria (Staphylococcus aureus). The results show that MRGO sheets could capture bacteria and kill them through an HFMF, showing a great potential in magnetic separation and antibacterial application. [ABSTRACT FROM AUTHOR]

In this study, a novel hemocompatible coating on stainless steel substrates was prepared by electrochemically copolymerizing 3,4-ethylenedioxythiophene (EDOT) with graphene oxide (GO), polystyrene sulfonate (PSS), or heparin (HEP) on SUS316L stainless steel, producing an anti-fouling (anti-protein adsorption and anti-platelet adhesion) surface to avoid the restenosis of blood vessels. The negative charges of GO, PSS, and HEP repel negatively charged proteins and platelets to achieve anti-fouling and anti-clotting. The results show that the anti-fouling capability of the poly(3,4-ethylenedioxythiophene) (PEDOT)/PSS coating is similar to that of the PEDOT/HEP coating. The anti-fouling capability of PEDOT/GO is higher than those of PEDOT/HEP and PEDOT/PSS. The reason for this is that GO exhibits negatively charged functional groups (COO−). The highest anti-fouling capability was found with the PEDOT/GO/HEP coating, indicating that electrochemical copolymerization of PEDOT with GO and HEP enhances the anti-fouling capability. Furthermore, the biocompatibility of the PEDOT coatings was tested with 3T3 cells for 1–5 days. The results show that all PEDOT composite coatings exhibited biocompatibility. The blood clotting time (APTT) of PEDOT/GO/HEP was prolonged to 225 s, much longer than the 40 s of pristine SUS316L stainless steel (the control), thus greatly improving the anti-blood-clotting capability of cardiovascular stents. [ABSTRACT FROM AUTHOR]

Repetitive movements at a constant rate require the integration of internal time counting and motor neural networks. Previous studies have proved that humans can follow short durations automatically (automatic timing) but require more cognitive efforts to track or estimate long durations. In this study, we studied sensorimotor oscillatory activities in healthy subjects and chronic stroke patients when subjects were performing repetitive finger movements. We found the movement-modulated changes in alpha and beta oscillatory activities were decreased with the increase of movement rates in finger lifting of healthy subjects and the non-paretic hands in stroke patients, whereas no difference was found in the paretic-hand movements at different movement rates in stroke patients. The significant difference in oscillatory activities between movements of non-paretic hands and paretic hands could imply the requirement of higher cognitive efforts to perform fast repetitive movements in paretic hands. The sensorimotor oscillatory response in fast repetitive movements could be a possible indicator to probe the recovery of motor function in stroke patients. [ABSTRACT FROM AUTHOR]