In the era of big data, HBase has been widely used in many business areas due to its good performance of massive data storage and management. Unfortunately, the native HBase only optimizes index for rowkey without creating indexes to the non-key column. A full table scan has to be used when querying the non-key column data, which greatly affects the efficiency of complex condition queries. In this paper, we present CSIndex, which is a coprocessor-based classified secondary index mechanism in HBase. CSIndex proposes the Observer-based secondary index management model to ensure the colocation of relevant data. Furthermore, according to the different data characteristics and query requirements, CSIndex designs the classified memory index model to balance query performance and index performance. On this basis, CSIndex proposes the Endpoint-based parallel query algorithm to reduce data transmission overhead, which improves query performance effectively. Finally, experiments are conducted on real datasets of vehicle trajectory. The results show that the query performance of CSIndex is significantly improved compared with the Solr-based scheme and HiBase, and has better overall performance.

Nitrogen starvation can induce cellular triacylglycerol (TAG) accumulation in different organisms with an unclear mechanism. In this study, we performed nutrient starvation and lipid droplet (LD) proteomics analyses of the filamentous fungus Metarhizium robertsii. Our results indicated that nitrogen starvation activated cell autophagic activity but inhibited the internalization of LDs into vacuoles for degradation. LD proteomic analyses identified an array of differentially accumulated proteins including autophagy-related (ATG) proteins, heat shock proteins, TAG metabolic and phospholipid biosynthetic enzymes when the fungus was grown in different nutrient conditions. In contrast to the highly activated MrATG8, the ATG proteins involved in vacuolar LD internalization were down-regulated after nitrogen starvation. Cellular TAG contents were increased in different ATG-gene null mutants of M. robertsii. In addition, TAG increase could be due to the up-regulation of TAG biogenesis along with the down-regulation of TAG catabolic enzymes in fungal cells after nitrogen deprivation. The data of this study benefit our understanding of the mechanism of nitrogen starvation induced TAG increase in different cells.

Phosphatidylcholine (PC) plays an important role in maintaining membrane integrity and functionality. In this study, two key genes (Mrpct and Mrpem) putatively involved in the cytidine diphosphate (CDP)-choline and phosphatidylethanolamine N-methyltransferase (PEMT) pathways for PC biosynthesis were characterized in the insect pathogenic fungus Metarhizium robertsii. The results indicated that disruption of Mrpct did not lead to any reduction of total PC content but impaired fungal virulence and increased cellular accumulation of triacylglycerol. Deletion of Mrpem reduced PC content and impaired fungal conidiation and infection structure differentiation but did not result in virulence defects. Lipidomic analysis revealed that deletion of Mrpct and Mrpem resulted in dissimilar effects on increase and decrease of PC moieties and other phospholipid species accumulations. Interestingly, we found that these two genes played opposite roles in activation of cell autophagy when the fungi were grown in a nutrient-rich medium. The connection between PC metabolism and autophagy was confirmed because PC content was drastically reduced in Mratg8Δ and that the addition of PC could rescue null mutant sporulation defect. The results of this study facilitate the understanding of PC metabolism on fungal physiology.

In this paper, a robot, namely iLeg, is designed for the purpose of rehabilitation of patients with hemiplegia or paraplegia. The iLeg is composed of one reclining seat and two leg orthoses, and each leg orthosis has three degrees of freedom, which correspond to the hip, knee, and ankle. Based on this robotic system, two controllers, i.e., passive training controller and active training controller, are proposed. The former takes advantage of the proportional-integral control method to solve the trajectory tracking problem, and the latter employs the surface electromyography signals to achieve active training. Two simplified impedance controllers, i.e., damping-type velocity controller and spring-type position controller, are designed for active training. A perceptron neural network detects movement intentions. The performance of the controllers was investigated with one able-bodied male. The results showed that the leg orthosis tracked the predefined trajectory based on the passive training controller, with the error rates of 0.45%, 0.44%, and 0.27%, respectively, for the hip, knee, and ankle. The active training controller whose loop rate is 6.67 Hz can move the leg orthosis smoothly, and the average recognition error of the perceptron neural network is less than 5%.

To the editor: Muscle wasting occurs with aging and in a wide range of catabolic diseases, such as cancer, diabetes, chronic renal disease, and heart failure, which can lead to a dramatically reduced quality of life and increased disease mortality. Several treatments are currently in development as a result of our understanding of the cellular mechanisms that cause muscle wasting.1Cohen S Nathan JA Goldberg AL Muscle wasting in disease: molecular mechanisms and promising therapies.Nat Rev Drug Discov. 2015; 14: 58-74Crossref PubMed Scopus (639) Google Scholar Myostatin, encoded by the MSTN gene, is a well-known cytokine that is activated in most disease conditions associated with muscle wasting, including cancer. Myostatin is primarily secreted by skeletal muscle cells and acts mainly through an autocrine/paracrine mode by activating the SMAD2/SMAD3 signaling pathway that subsequently triggers transcription programs leading to muscle atrophy.1Cohen S Nathan JA Goldberg AL Muscle wasting in disease: molecular mechanisms and promising therapies.Nat Rev Drug Discov. 2015; 14: 58-74Crossref PubMed Scopus (639) Google Scholar,2Lee YS Huynh TV Lee SJ Paracrine and endocrine modes of myostatin actin.J Appl Physiol 1985. 2016; 120: 592-598Crossref PubMed Scopus (29) Google Scholar Inhibition of myostatin has been identified as a promising strategy to alleviate muscle wasting, especially given that loss of functional myostatin in humans is not associated with apparent deleterious effects other than muscle hypertrophy.3Schuelke M Wagner KR Stolz LE Hubner C Riebel T Komen W et al.Myostatin mutation associated with gross muscle hypertrophy in a child.N Engl J Med. 2014; 350: 2682-2688Crossref Scopus (1094) Google Scholar Monoclonal antibodies that bind to myostatin to inhibit its function are currently in clinical trials as a potential therapy to treat cachexia syndrome in cancer patients.1Cohen S Nathan JA Goldberg AL Muscle wasting in disease: molecular mechanisms and promising therapies.Nat Rev Drug Discov. 2015; 14: 58-74Crossref PubMed Scopus (639) Google Scholar,4Ebner N Steinbeck L Doehner W Anker SD von Heahling S Highlights from the 7th Cachexia Conference: muscle wasting pathophysiological detection and novel treatment strategies.J Cachexia Sarcopenia Muscle. 2014; 5: 27-34Crossref PubMed Scopus (52) Google Scholar However, the nature of antibody-based drugs means that this strategy requires repeated injections chronically. With the emergence of different genome-editing tools that enable investigators to introduce genetic alterations into specific genomic sites at will, the use of genome editing for human therapeutic applications is being explored in a variety of preclinical models of diseases.5Maeder ML Gersbach CA Genome-editing technologies for gene and cell therapy.Mol Ther. 2016; 24: 475-487Abstract Full Text Full Text PDF PubMed Scopus (405) Google Scholar Recently, in vivo genome editing targeting muscle tissues with clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems showed efficacy in a mouse model of Duchenne muscular dystrophy.6Long C Amoasii L Mireault AA McAnally JR Li H Sanchez-Ortiz E et al.Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy.Science. 2016; 351: 400-403Crossref PubMed Scopus (671) Google Scholar,7Nelson CE Hakim CH Ousterout DG Thakore PI Moreb EA Castellanos Rivera RM et al.In vivo genome editing improves muscle function in a mouse model of Duchenne muscular dystrophy.Science. 2016; 351: 403-407Crossref PubMed Scopus (806) Google Scholar,8Tabebordbar M Zhu K Cheng JK Chew WL Widrick JJ Yan WX et al.In vivo gene editing in dystrophic mouse muscle and muscle stem cells.Science. 2016; 351: 407-411Crossref PubMed Scopus (741) Google Scholar With a view to establishing a novel therapeutic modality toward muscle-wasting syndrome, we used CRISPR/Cas9 to directly target Mstn in vivo in skeletal muscle cells to prevent loss of muscle mass. We reasoned that the ability to disrupt Mstn and block the myostatin pathway in some, if not all, skeletal muscle cells would be sufficient to partially prevent atrophy of targeted cells and its neighbor cells, thus preserving muscle function to some extent. To perform in vivo targeting of skeletal muscle cells, we adapted a Staphylococcus aureus (SaCas9)/adeno-associated virus 8 (AAV8) system9Ran FA Cong L Yan WX Scott DA Gootenberg JS Kriz AJ et al.In vivo genome editing using Staphylococcus aureus Cas9.Nature. 2014; 520: 186-191Crossref Scopus (1769) Google Scholar for specific muscle cell targeting by using a tissue-specific double muscle creatine kinase (dMCK) promoter (∼500 bp)10Wang B Li J Fu FH Chen C Zhu X Zhou L et al.Construction and analysis of compact muscle-specific promoters for AAV vectors.Gene Ther. 2008; 15: 1489-1499Crossref PubMed Scopus (97) Google Scholar to drive SaCas9 expression (Figure 1a). Efficient expression of genes driven by the dMCK promoter was confirmed by in vivo luminescence examinations with mice injected with AAV8/dMCK promoter-GFP-2a-luciferase viruses for up to two months (data not shown). We next screened candidate SaCRISPR guide RNAs (gRNAs) targeting the first exon of the mouse Mstn gene in mouse NIH-3T3 cells. Among the three gRNAs we tested, we found that one gRNA (guide-2, targeting sequences: GGGCTGTGTAATGCATGTGCG+TGGAG) displayed a ∼50% mutagenesis rate at the on-target site as judged by Surveyor assays (Figure 1b). We further accessed the potential off-target mutagenesis at ten sites that are mostly matched to the guide-2 targeting sequences by genome-wide prediction using the CRISPR Design Server (http://crispr.mit.edu/) (Supplementary Table S1 online). Although one of the sites displayed a relatively high score (5.0) with mismatches by two bases in protospacer matching sequences, lack of protospacer-adjacent motif prevented efficient targeting at this locus. With the limit of detection by the Surveyor assays in NIH-3T3 cells, we found no evidence of significant off-target mutagenesis (Figure 1c). We thus used guide-2 for subsequent in vivo targeting. We made an AAV for efficient delivery of SaCas9 and this gRNA (AAV-SaCRISPR-Mstn) to muscle cells in vivo, using an AAV expressing GFP-2a-luciferase (AAV-GFP-2a-Luci) as a control. To test the hypothesis that genome editing could disrupt Mstn in vivo in muscle tissues and thus prevent muscle loss in catabolic conditions, we administered the AAV-SaCRISPR-Mstn and the AAV-GFP-2a-Luci viruses locally to gastrocnemius muscles (4–5 × 1010 viral particles per injection and six to eight injections per limb) in six-week-old male wild-type C57BL/6 mice, with six mice in each group. We waited around four weeks to allow sufficient expression of SaCas9 in vivo, after which we injected Lewis lung carcinoma cells (3 × 106 cells per mouse) subcutaneously into all mice to induce cancer-associated muscle loss. The mice were then analyzed at the end of week 8 (Figure 1d). Immunostainings of gastrocnemius muscles from AAV-GFP-2a-Luci virus-treated animals showed a mosaic pattern of GFP+ myofibers (Supplementary Figure S1a), indicating efficient but uneven infections by local viral injection. Surveyor analysis in muscle tissues displayed around 5% mutagenesis at the Mstn gene locus in four out of six CRISPR-treated mice, with no evidence of mutagenesis in control mice (Figure 1e). Furthermore, deep sequencing of gRNA targeting sites indicated 4.5 to 8.2% (5.8% on average) indel formation at the target sites in muscle tissues receiving AAV-SaCRISPR-Mstn in all six mice, in contrast to around 0.2% indel formation in muscle tissues receiving AAV-GFP-2a-Luci viruses (Figure 1f). We therefore also included the two mice that showed little editing activity in Surveyor assays (Figure 1e) for all functional analyses. We observed a consistent growth of tumors (as judged by tumor volume measurement) and reduction of fat mass (as judged by NMR analysis) in both groups after tumor cell engraftment (Figure 2a) and a significant decrease of tumor-free body weight (∼9%) at the end point (week 8) in comparison to week 0 in both groups (Figure 2b), indicating the development of cancer-associated cachexia; no statistically significant difference was observed in the above measurements between AAV-SaCRISPR-Mstn–treated and control mice. Next, we analyzed muscle conditions in mice from different groups, including another group of mice with no tumor engraftment and no virus treatment as healthy controls (designated as "tumor-free control"). No significant differences of myostatin concentration in mouse serum were noticed as a result of the relatively low mutagenesis efficiency (Figure 2c). However, we observed a small but significant ∼9% increase of gastrocnemius and soleus muscle weight in the CRISPR-treated group compared to the AAV-GFP-2a-Luci group (Figure 2d). We suspected that this was a result of the major autocrine/paracrine-working mode of myostatin2Lee YS Huynh TV Lee SJ Paracrine and endocrine modes of myostatin actin.J Appl Physiol 1985. 2016; 120: 592-598Crossref PubMed Scopus (29) Google Scholar; disruption of Mstn gene in a few myotubes caused a decrease in local myostatin concentration, leading to attenuated myostatin signaling pathway in CRISPR-targeted cells as well as its neighbor cells, and thus alleviated atrophy of these myotubes. To further test this hypothesis, we also measured the grip strength of virus-treated limbs. Compared to tumor-free mice, mice carrying tumors displayed a dramatic reduction in grip strength as cachexia developed (Figure 2e). However, we observed a significant improvement (>25%) in grip strength in mice treated with AAV-SaCRISPR-Mstn in comparison to AAV-GFP-2a-Luci, indicating partially recovered muscle function in CRISPR-treated mice (Figure 2e). Moreover, CRISPR-treated mice showed clearly ameliorated atrophy of myocytes, as indicated by hematoxylin and eosin staining of muscle fibers (Figure 2f). Interestingly, we found coexistence of myotubes that with severe atrophy (indicated by dotted-line boxes) and with alleviated atrophy (indicated by full-line boxes) in a single slide from CRISPR-treated mice (Supplementary Figure S1b), suggesting that alleviated muscle atrophy is mostly likely a result of decreased local myostatin production instead of a systematic reduction in blood myostatin concentration. In summary, we demonstrated in this study that AAV8-mediated delivery of SaCas9 driven by a muscle cell–specific promoter can be used to disrupt Mstn in mouse muscle in vivo. Although the overall efficiency of genome editing in muscle tissues was low (around 5% as judged by Surveyor assay and deep sequencing), consistent with previous studies with in vivo muscle targeting,6Long C Amoasii L Mireault AA McAnally JR Li H Sanchez-Ortiz E et al.Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy.Science. 2016; 351: 400-403Crossref PubMed Scopus (671) Google Scholar, 7Nelson CE Hakim CH Ousterout DG Thakore PI Moreb EA Castellanos Rivera RM et al.In vivo genome editing improves muscle function in a mouse model of Duchenne muscular dystrophy.Science. 2016; 351: 403-407Crossref PubMed Scopus (806) Google Scholar, 8Tabebordbar M Zhu K Cheng JK Chew WL Widrick JJ Yan WX et al.In vivo gene editing in dystrophic mouse muscle and muscle stem cells.Science. 2016; 351: 407-411Crossref PubMed Scopus (741) Google Scholar disruption of myostatin function in a portion of muscle cells was shown to be sufficient to reduce cachexia in targeted cells or neighbor cells due to the major autocrine/paracrine working mode of myostatin, thus preserving muscle function (>25% as judged by grip-strength measurement and ameliorated atrophy) in cancer-induced cachexia syndrome. As a proof of concept, our study raises the possibility that a patient could benefit with alleviated muscle-wasting syndrome induced by cancer or other catabolic conditions through the use of CRISPR/Cas9-mediated targeting of the MSTN gene in skeletal muscle tissues, although continued development of this strategy to improve the targeting efficacy and assessment of safety will be needed before it can be realized in human beings. Download .pdf (.12 MB) Help with pdf files Table S1. Top ten genome-wide off-target sites used in this study. Download .pdf (.23 MB) Help with pdf files Figure S1. Effects of targeting the Mstn gene in skeletal muscle cells in mice with cancer-associated muscle atrophy. We thank Kiran Musunuru (University of Pennsylvania) and Yan Chen (Institute for Nutritional Sciences, SIBS, China) for helpful comments, suggestions, and critical reading of this manuscript. We thank Xianfeng Chen for assistance in data analysis and Junjie Xiao (Shanghai University, China) for help in measuring mouse-grip strength. This work was supported by grants from Ministry of Science and Technology of the People's Republic of China (2016YFC1304900), the Shanghai Institutes for Biological Sciences Fellowship (Y5Y1X41491) (Y.Z.), the National Natural Science Foundation of China (81500614 to Y.Z.), the Hundred Talents Program of the Chinese Academy of Sciences (Q.D.), the National Youth 1000 Talents Program (Q.D.), the Shanghai Pujiang Program (15PJ1409200 to Q.D.), and the National Natural Science Foundation of China (31670829 to Q.D.).

目的：心肌梗死后及时再灌注是挽救缺血心肌必需的步骤，但该过程伴随着再灌注损伤。缺血／再灌注（ I／R）造成的线粒体Ca2＋（［Ca2＋］m）超载、活性氧（reactive oxygenspecies， ROS）大量释放以及线粒体膜通透性通道（mitochondrial permeability transition pore， MPTP），并导致线粒体严重的结构和功能破坏，是心肌细胞I／R损伤的主要原因。因此寻找保护线粒体的关键靶点，对于深入理解I／R导致心肌细胞结构和功能损伤机制，进而提出心肌保护的新策略至关重要。本实验旨在探讨多种心肌保护措施对I／R心肌的线粒体保护作用及其机制。方法：利用大鼠离体心脏全心I／R模型和成体心肌细胞模拟I／R模型，结合基因的遗传学操作，探讨间歇性低压低氧（ intermittent hypobaric hypoxia， IHH）和过氧化氢预处理（ hydrogen peroxide pre－conditioning， H2 O2 PC）对心肌线粒体的保护作用。利用Rhod－2、MitoSOX和TMRE 荧光染料分别实时监测线粒体［ Ca2＋］ m 超载、线粒体ROS的释放和线粒体膜电位（ mitochondrial membrane potential，ΔΨm ）的变化。分离缺血前和再灌注后活体心肌组织的线粒体，检测线粒体膜电位、耗氧率、ROS释放量及三磷酸腺苷（ adenosine triphosphate， ATP）合成酶活性。结果：IHH可以通过ROS降低［ Ca2＋］ m 超载，改善心肌I／R损伤后线粒体的膜电位、耗氧率、ATP合成酶活性及心肌ATP含量的降低，进而改善心肌I／R后心功能，减少心肌梗死面积。此外，H2 O2 PC在I／R过程中通过UCP3降低［ Ca2＋］ m 超载及防止ΔΨm 丧失，进而改善心肌I／R后心功能，减少心肌梗死面积。本研究揭示了线粒体在心肌I／R损伤和保护中的重要地位和调控机制，为解释IHH和H2 O2 PC心肌保护作用的机理提供新视角，并为缺血性心脏病的临床治疗提供新的实验证据。

ABSTRACT Two-component signaling pathways generally include sensor histidine kinases and response regulators. We identified an ortholog of the response regulator protein Skn7 in the insect-pathogenic fungus Metarhizium robertsii, which we named MrSkn7. Gene deletion assays and functional characterizations indicated that MrSkn7 functions as a transcription factor. The MrSkn7 null mutant of M. robertsii lost the ability to sporulate and had defects in cell wall biosynthesis but was not sensitive to oxidative and osmotic stresses compared to the wild type. However, the mutant was able to produce spores under salt stress. Insect bioassays using these spores showed that the virulence of the mutant was significantly impaired compared to that of the wild type due to the failures to form the infection structure appressorium and evade host immunity. In particular, deletion of MrSkn7 triggered cell autolysis with typical features such as cell vacuolization, downregulation of repressor genes, and upregulation of autolysis-related genes such as extracellular chitinases and proteases. Promoter binding assays confirmed that MrSkn7 could directly or indirectly control different putative target genes. Taken together, the results of this study help us understand the functional divergence of Skn7 orthologs as well as the mechanisms underlying the development and control of virulence in insect-pathogenic fungi.

Moderate enhanced reactive oxygen species (ROS) during early reperfusion trigger the cardioprotection against ischemia/reperfusion (I/R) injury, while the mechanism is largely unknown. Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) contributes to the cardioprotection but whether it is activated by ROS and how it regulates Ca(2+) homeostasis remain unclear. Here we investigated whether the ROS generated during early reperfusion protect the heart/cardiomyocyte against I/R-induced Ca(2+) overload and contractile dysfunction via the activation of JAK2/STAT3 signaling pathway by using a cardioprotective model of intermittent hypobaric hypoxia (IHH) preconditioning. IHH improved the postischemic recovery of myocardial contractile performance in isolated rat I/R hearts as well as Ca(2+) homeostasis and cell contraction in simulated I/R cardiomyocytes. Meanwhile, IHH enhanced I/R-increased STAT3 phosphorylation at tyrosine 705 in the nucleus and reversed I/R-suppressed STAT3 phosphorylation at serine 727 in the nucleus and mitochondria during reperfusion. Moreover, IHH improved I/R-suppressed sarcoplasmic reticulum (SR) Ca(2+)-ATPase 2 (SERCA2) activity, enhanced I/R-increased Bcl-2 expression, and promoted the co-localization and interaction of Bcl-2 with SERCA2 during reperfusion. These effects were abolished by scavenging ROS with N-(2-mercaptopropionyl)-glycine (2-MPG) and/or by inhibiting JAK2 with AG490 during the early reperfusion. Furthermore, IHH-improved postischemic SERCA2 activity and Ca(2+) homeostasis as well as cell contraction were reversed after Bcl-2 knockdown by short hairpin RNA. In addition, the reversal of the I/R-suppressed mitochondrial membrane potential by IHH was abolished by 2-MPG and AG490. These results indicate that during early reperfusion the ROS/JAK2/STAT3 pathways play a crucial role in (i) the IHH-maintained intracellular Ca(2+) homeostasis via the improvement of postischemic SERCA2 activity through the increase of SR Bcl-2 and its interaction with SERCA2; and (ii) the IHH-improved mitochondrial function.

d-Xylose is the most abundant fermentable pentose in nature and can serve as a carbon source for many bacterial species. Since d-xylose constitutes the major component of hemicellulose, its metabolism is important for lignocellulosic biomass utilization. Here, we report a six-protein module for d-xylose signaling, uptake and regulation in solvent-producing Clostridium beijerinckii. This module consists of a novel three-component system' (a putative periplasmic ABC transporter substrate-binding protein XylFII and a two-component system LytS/YesN) and an ABC-type d-xylose transporter XylFGH. Interestingly, we demonstrate that, although XylFII harbors a transmembrane domain, it is not involved in d-xylose transport. Instead, XylFII acts as a signal sensor to assist the response of LytS/YesN to extracellular d-xylose, thus enabling LytS/YesN to directly activate the transcription of the adjacent xylFGH genes and thereby promote the uptake of d-xylose. To our knowledge, XylFII is a novel single transmembrane sensor that assists two-component system to respond to extracellular sugar molecules. Also of significance, this three-component system' is widely distributed in Firmicutes, indicating that it may play a broad role in this bacterial phylum. The results reported here provide new insights into the regulatory mechanism of d-xylose sensing and transport in bacteria.

AIMS:Uncoupling protein 3 (UCP3), located in the mitochondrial inner membrane, is cardioprotective, but its mechanisms of preserving mitochondrial function during ischaemia/reperfusion (I/R) are not fully understood. This study investigated whether UCP3 mediates/mimics the cardioprotection of H₂O₂ preconditioning (H₂O₂PC) against I/R injury and the downstream pathway that mediates H₂O₂PC- and UCP3-afforded cardioprotection.METHODS AND RESULTS:H₂O₂PC at 20 µM for 5 min significantly improved post-ischaemic functional recovery and reduced lactate dehydrogenase (LDH) release and infarct size with concurrently up-regulated UCP3 expressions in perfused rat hearts subjected to global no-flow I/R. These protections were blocked by UCP3 knockdown with short hairpin RNA but mimicked by UCP3 overexpression. Consistently, H₂O₂PC-attenuated I/R-induced cytosolic and mitochondrial Ca(2+) overload, Ca(2+) transient suppression, mitochondrial reactive oxygen species burst, and loss of mitochondrial inner membrane potential were reversed by UCP3 knockdown but mimicked by UCP3 overexpression. Moreover, co-immunoprecipitation assay revealed an interaction of UCP3 with the mitochondrial permeability transition pore (mPTP) component, adenine nucleotide translocator (ANT), while the cardioprotection induced by H₂O₂PC- and UCP3 overexpression in mitochondria, cardiac function, and cell survival was abolished by atractyloside, a mPTP opener binding to ANT, and partially inhibited by a PI3K/Akt inhibitor wortmannin. Furthermore, H₂O₂PC up-regulated the phosphorylation of Akt, and glycogen synthase kinase 3β was blocked by UCP3 knockdown but mimicked by UCP3 overexpression.CONCLUSION:UCP3 mediates the cardioprotection of H₂O₂PC against I/R injury by preserving the mitochondrial function through inhibiting mPTP opening via the interaction with ANT and the PI3K/Akt pathway. Our findings reveal novel mechanisms of UCP3 in the cardioprotection.

Bax inhibitor 1 (BI-1) is a highly conserved protein originally identified as a suppressor of the proapoptotic protein Bax to inhibit cell death in animals and plants. The orthologs of BI-1 are widely distributed in filamentous fungi but their functions remain largely unknown. Herein, we report the identification and characterizations of MrBI-1, an ortholog of BI-1, in the entomopathogenic fungus Metarhizium robertsii. First, we found that MrBI-1 could partially rescue mammalian Bax-induced cell death in yeast. Deletion of MrBI-1 impaired fungal development, virulence and heat tolerance in M. robertsii. We also demonstrated that inactivation of MrBI-1 reduced fungal resistance to farnesol but not to hydrogen peroxide, suggesting that MrBI-1 contributes to antiapoptotic-like cell death via the endoplasmic reticulum stress-signaling pathway rather than the classical mitochondrium-dependent pathway. In particular, we found that unlike the observations in yeasts and plants, expression of mammalian Bax did not lead to a lethal effect in M. robertsii; however, it did aggravate the fungal apoptotic effect of farnesol. The results of this study advance our understanding of BI-1-like protein functions in filamentous fungi.

This paper is a continuation of previous published work by the same authors on Personalized Modelling and Evolving Spiking Neural Network Reservoir architecture (PMeSNNr). The focus is on improvement of predictive modeling methods for the stroke occurrences case study utilizing an enhanced NeuCube architecture. The adaptability of the new architecture leads towards understanding feature correlations that affect the outcome of the study and extracts new knowledge from hidden patterns that reside within the associations. Through this new method, estimation of the earliest time point for stroke prediction is possible. This study also highlighted the improvement from designing a new experimental dataset compared to previous experiments. Comparative experiments were also carried out using conventional machine learning algorithms such as kNN, wkNN, SVM and MLP to prove that our approach can result in much better accuracy level.

The paper presents a novel method and system for personalised (individualised) modelling of spatio/spectro-temporal data (SSTD) and prediction of events. A novel evolving spiking neural network reservoir system (eSNNr) is proposed for the purpose. The system consists of spike-time encoding module of continuous value input information into spike trains; a recurrent 3D SNNr; eSNN as an evolving output classifier. Such system is generated for every new individual, using existing data of similar individuals. Subject to proper training and parameter optimisation, the system is capable of accurate spatio-temporal pattern recognition (STPR) and of early prediction of individual events. The method and the system are generic, applicable to various SSTD and classification and prediction problems. As a case study, the method is applied for early prediction of occurrence of stroke on an individual basis. Preliminary experiments demonstrated a significant improvement in accuracy and time of event prediction when using the proposed method when compared with standard machine learning methods, such as MLR, SVM, and MLP. Future development and applications are discussed.

A lower limb rehabilitation robot, namely iLeg, has been developed recently. Since active exercises have been proven to be effective for neurorehabilitation and motor recovery, they are suggested to be implemented on iLeg. To this goal, patients' motion intention should be recognized. Therefore, a method based on the dynamic model of the human-robot interface (HRI) is designed to recognize the human motion intention. This paper is devoted to modeling and identifying the dynamics of the HRI. Firstly, the dynamic model of the HRI is designed by combining the dynamic models of the human leg and iLeg, where the human leg dynamic model (HLDM) is mainly concerned. By considering the motion trajectories during the rehabilitation exercises provided by iLeg, the human leg can be taken as a manipulator with two degrees of freedom; meanwhile, the joint angles and torques of the human leg can be measured indirectly by using the position and torque sensors mounted on the joints of iLeg. As a result, an 8-parameter HLDM can be designed by using the Lagrangian method. Then, the dynamic model of the HRI is identified by respectively and independently identifying the undetermined dynamic parameters of iLeg and the HLDM, where the dynamic parameters of the HLDM are mainly considered. Finally, the feasibility of the dynamic model of the HRI is validated by experiments.

To design a control strategy for iLeg, an exoskeleton robot developed for lower limb rehabilitation aiming at investigating the feasibility of integrating functional electrical stimulation (FES) with robot-based rehabilitation training, an FES-assisted training strategy combined with impedance control, has been proposed in this paper. Through impedance control, an active compliance of the robot is established, and the patient’s voluntary effort to accomplish the training task is inspired. During the training process, the patient’s related muscles are applied with FES which provides an extra assistance to the patient. The intensity of the FES is properly chosen in order to induce a desired active torque which is proportional to the voluntary effort extracted from the electromyography signals of the related muscles using back propagation neural networks. This kind of enhancement serves as a positive feedback which reminds the patient of the correct attempt to fulfill the desired motion. FES control is conducted by a combination of neural network-based feedforward controller and a PD feedback controller. Simulation conducted using Matlab and the experiment with a spinal cord injury subject and a healthy subject have shown satisfactory results which verify the feasibility of this control strategy.

The number of paralytic sufferers is currently growing huge and the rehabilitation for them is usually a long-time process. Compared to the traditional physiotherapy, rehabilitation with the assistance of robots can reduce the cost and time, and less labor intensity is required. Moreover, various training strategies are provided by robots, so that rehabilitation effect can be improved. Lower limb rehabilitation robots are categorized into horizontal exercisers, vertical locomotors, sit-to-stand aids and multi-orientation hybrids, according to the posture of patient during therapy. Horizontal exercisers are subcategorized into end effectors and exoskeletons, and vertical locomotors are further grouped as suspending body weight support (sBWS) based gait trainers and stand-alone wearables. Interactive control between mechanism and patient is required to create a secure, comfortable and natural training environment for paralytic patients. According to the signals employed to deduce the movement intention of patients, interactive control methods are classified into force-based control and biomedical-signal-based control. Two approaches that are in particular worth mentioning for force-based interactive control are hybrid force-position control and impedance control. Surface electromyogram (sEMG) and electroencephalogram (EEG) are two mostly used signals for biomedical-signal-based control.

Inhibition of matrix metalloproteinases-2 (MMP-2) activation renders cardioprotection from ischemia/reperfusion (I/R) injury; however, the signaling pathways involved have not been fully understood. Intermittent hypobaric hypoxia (IHH) has been shown to enhance myocardial tolerance to I/R injury via triggering intrinsic adaptive responses. Here we investigated whether IHH protects the heart against I/R injury via the regulation of MMP-2 and how the MMP-2 is regulated. IHH (Po2 = 84 mmHg, 4-h/day, 4 wk) improved postischemic myocardial contractile performance, lactate dehydrogenase (LDH) release, and infarct size in isolated perfused rat hearts. Moreover, IHH reversed I/R-induced MMP-2 activation and release, disorders in the levels of MMP-2 regulators, peroxynitrite (ONOO(-)) and tissue inhibitor of metalloproteinase-4 (TIMP-4), and loss of the MMP-2 targets α-actinin and troponin I. This protection was mimicked, but not augmented, by a MMP inhibitor doxycycline and lost by the α1-adrenoceptor (AR) antagonist prazosin. Furthermore, IHH increased myocardial α1A-AR and α1B-AR density but not α1D-AR after I/R. Concomitantly, IHH further enhanced the translocation of PKC epsilon (PKCε) and decreased the release of mitochondrial cytochrome c due to I/R via the activation of α1B-AR but not α1A-AR or α1D-AR. IHH-conferred cardioprotection in the postischemic contractile function, LDH release, MMP-2 activation, and nitrotyrosine as well as TIMP-4 contents were mimicked but not additive by α1-AR stimulation with phenylephrine and were abolished by an α1B-AR antagonist chloroethylclonidine and a PKCε inhibitor PKCε V1-2. These findings demonstrate that IHH exerts cardioprotection through attenuating excess ONOO(-) biosynthesis and TIMP-4 loss and sequential MMP-2 activation via the activation of α1B-AR/PKCε pathway.

This paper proposes a novel leg orthosis for lower limb rehabilitation robots of the sitting/lying type. It consists of three joint mechanisms: hip, knee and ankle, and two sets of links: thigh and crus. Each driving motor is located close to the associated joint and the rotational axis of each joint mechanism is unique and stable. These features make it outperform the similar mechanisms in stability and dynamic performance. Different forms of eccentric slider-crank mechanisms are applied in the three joint mechanisms, respectively, such that they can be optimized independently. The optimization problems for the hip and knee joint mechanisms, characterized as strongly nonlinear, are developed respectively. Then, a particle swarm optimization algorithm is used to obtain the optimal solutions, which are subsequently validated by comprehensive comparisons. Moreover, the kinematics necessary for motion control and trajectory tracking are investigated, which denote the relationships between the displacements and velocities of the joint mechanisms, lead screws and the end effector. Finally, this paper illustrates the feasibility of the application of the leg orthosis to actual rehabilitation exercises by a simulation example.

Fungal polyketide synthases (PKSs) and their related gene clusters are highly diversified at both inter- and intra-specific levels. The most well characterized PKS enzymes include those responsible for the biosynthesis of polyketide pigments such as melanins. The genome of the insect pathogenic fungus Metarhizium robertsii contains 20 type I PKSs but none has been functionally characterized. In this study, two PKS genes (designated as MrPks1 and MrPKs2) showing homologies to those counterparts for the biosynthesis of heptaketide pigments and dihydroxynaphthalene (DHN)-melanins, respectively, were deleted in two different strains of M. robertsii. The results indicated that disruption of MrPks1 but not MrPks2 impaired fungal culture pigmentation and cell wall structure. In addition to the negative effect of the DHN-melanin pathway inhibitor, it was postulated that DHN-melanin would not be produced by M. robertsii. Various assays revealed that the stress resistance abilities against ultraviolet radiation, heat shock and oxidants, as well as virulence against insects were not impaired in ΔMrPks1 and ΔMrPks2 isolates when compared with the wild-type strain. Thus, the non-melanin pigment(s) produced by the fungus do not contribute to cell damage protection and pathogenicity in M. robertsii. Physiological differences were evident in the two examined wild-type strains. The results from this study advance the understanding of functional divergence of fungal PKSs.