The Arabidopsis WRR4A and WRR4B paralogous NLR proteins both confer recognition of multiple Albugo candida effectors.

The oomycete Albugo candida causes white blister rust, an important disease of Brassica crops. Distinct races of A. candida are defined by their capacity to infect different host plant species. Each A. candida race encodes secreted proteins with a CX2 CX5 G ('CCG') motif that are polymorphic and show presence/absence variation, and are therefore candidate effectors. The White Rust Resistance 4 (WRR4) locus in Arabidopsis thaliana accession Col-0 contains three genes that encode intracellular nucleotide-binding domain leucine-rich repeat immune receptors. The Col-0 alleles of WRR4A and WRR4B confer resistance to multiple A. candida races, although both WRR4A and WRR4B can be overcome by the Col-0-virulent race 4 isolate AcEx1. Comparison of CCG candidate effectors in avirulent and virulent races, and transient co-expression of CCG effectors from four A. candida races in Nicotiana sp. or A. thaliana, revealed CCG effectors that trigger WRR4A- or WRR4B-dependent hypersensitive responses. We found eight WRR4A-recognised CCGs and four WRR4B-recognised CCGs, the first recognised proteins from A. candida for which the cognate immune receptors in A. thaliana are known. This multiple recognition capacity potentially explains the broad-spectrum resistance to several A. candida races conferred by WRR4 paralogues. We further show that of five tested CCGs, three confer enhanced disease susceptibility when expressed in planta, consistent with A. candida CCG proteins being effectors.

Emerging microfluidics-enabled platforms for osteoarthritis management: from benchtop to bedside.

Osteoarthritis (OA) is a prevalent debilitating age-related joint degenerative disease. It is a leading cause of pain and functional disability in older adults. Unfortunately, there is no cure for OA once the damage is established. Therefore, it promotes an urgent need for early detection and intervention of OA. Theranostics, combining therapy and diagnosis, emerges as a promising approach for OA management. However, OA theranostics is still in its infancy. Three fundamental needs have to be firstly fulfilled: i) a reliable OA model for disease pathogenesis investigation and drug screening, ii) an effective and precise diagnostic platform, and iii) an advanced fabrication approach for drug delivery and therapy. Meanwhile, microfluidics emerges as a versatile technology to address each of the needs and eventually boost the development of OA theranostics. Therefore, this review focuses on the applications of microfluidics, from benchtop to bedside, for OA modelling and drug screening, early diagnosis, and clinical therapy. We first introduce the basic pathophysiology of OA and point out the major unfilled research gaps in current OA management including lack of disease modelling and drug screening platforms, early diagnostic modalities and disease-modifying drugs and delivery approaches. Accordingly, we then summarize the state-of-the-art microfluidics technology for OA management from in vitro modelling and diagnosis to therapy. Given the existing promising results, we further discuss the future development of microfluidic platforms towards clinical translation at the crossroad of engineering and biomedicine.

An Assessment of China's Digital Trade Development and Influencing Factors.

The pattern and scale of commerce worldwide have been greatly transformed by the Fourth Industrial Revolution and technological advancement; digital trade has become the primary form of trade in the digital economy. On the basis of information network infrastructure, information technology level, digital industrialization level, and industrial digitalization level, this study establishes a comprehensive assessment system that applies an entropy-TOPSIS model to evaluate digital trade development level in China. The results indicate that digital trade in China was steadily growing between 2010 and 2019. A principal component analysis is conducted to identify factors affecting the digital trade development level in China. The analysis results suggest that Internet development, population income, industrial structure, payment convenience level, fixed asset investment, online transaction scale, and economic development all have positive effects on the digital trade development level in China, with payment convenience level having the greatest influence. By contrast, state intervention and degree of dependence on foreign trade have a negative effect on digital trade development.

Show me your ID: NLR immune receptors with integrated domains in plants.

Nucleotide-binding and leucine-rich repeat receptors (NLRs) are intracellular plant immune receptors that recognize pathogen effectors secreted into the plant cell. Canonical NLRs typically contain three conserved domains including a central nucleotide binding (NB-ARC) domain, C-terminal leucine-rich repeats (LRRs) and an N-terminal domain. A subfamily of plant NLRs contain additional noncanonical domain(s) that have potentially evolved from the integration of the effector targets in the canonical NLR structure. These NLRs with extra domains are thus referred to as NLRs with integrated domains (NLR-IDs). Here, we first summarize our current understanding of NLR-ID activation upon effector binding, focusing on the NLR pairs Pik-1/Pik-2, RGA4/RGA5, and RRS1/RPS4. We speculate on their potential oligomerization into resistosomes as it was recently shown for certain canonical plant NLRs. Furthermore, we discuss how our growing understanding of the mode of action of NLR-ID continuously informs engineering approaches to design new resistance specificities in the context of rapidly evolving pathogens.

High-throughput and label-free isolation of senescent murine mesenchymal stem cells.

Under internal or external insults such as aging and oxidative stresses, cells are induced into a senescent state and stop cellular division permanently. As senescent cells (SnCs) accumulate, the regeneration capacity of biological tissue would be compromised, which has been found to be associated with a plethora of age-related disorders. Therefore, isolating SnCs becomes necessary. To address the lack of effective surface markers for SnCs isolation, a label-free microfluidic device was proposed in this paper, in which a spiral microchannel was deployed to isolate SnCs based on their size differences. We adopted a well-received cellular senescence model by exerting excessive oxidative stress to murine mesenchymal stem cells. This model was then validated through a series of SnCs characterizations including size measurement, p16INK4a expression level, senescence-associated beta-galactosidase, and doubling time. The senescence chip demonstrated an efficiency of 75% and viability over 85% at a flow rate of 5 ml/min. The average cell size from the inner outlet was 5 µm larger than that from the outer outlet. The isolated cells had a sixfold higher p16INK4a expression level. Overall, the chip had an area under curve of 0.719 in the receiver operating characteristic analysis, showing decent performance in sorting SnCs. By having the ability to perform size-based sorting at a high flow rate, such a microfluidic device can provide high-throughput and label-free isolation of SnCs. To further improve the isolation performance, the device can be modified to introduce additional physical biomarkers of SnCs such as stiffness. This device poses a good potential in purification for cytotherapy or estimation of biological age.

Synthesis and in vitro opioid receptor functional antagonism of analogues of the selective kappa opioid receptor antagonist (3R)-7-hydroxy-N-((1S)-1-{[(3R,4R)-4-(3-hydroxyphenyl)-3,4-dimethyl-1-piperidinyl]methyl}-2-methylpropyl)-1,2,3,4-tetrahydro-3-isoquinolinecarboxamide (JDTic).

In previous structure-activity relationship (SAR) studies, we identified (3 R)-7-hydroxy- N-((1 S)-1-{[(3 R,4 R)-4-(3-hydroxyphenyl)-3,4-dimethyl-1-piperidinyl]methyl}-2-methylpropyl)-1,2,3,4-tetrahydro-3-isoquinolinecarboxamide (JDTic, 1) as the first potent and selective kappa opioid receptor antagonist from the trans-3,4-dimethyl-4-(3-hydroxyphenyl)piperidine class of opioid antagonist. In the present study, we report the synthesis and in vitro opioid receptor functional antagonism of a number of analogues of 1 using a [ (35) S]GTPgammaS binding assay. The results from the studies better define the pharmacophore for this class of kappa opioid receptor antagonist and has identified new potent and selective kappa antagonist. (3 R)-7-Hydroxy- N-[(1 S,2 S)-1-{[(3 R,4 R)-4-(3-hydroxyphenyl)-3,4-dimethylpiperidin-1-yl]methyl}-2-methylbutyl]-1,2,3,4-tetrahydroisoquinoline-3-carboxamide ( 3) with a K e value of 0.03 nM at the kappa receptor and 100- and 793-fold selectivity relative to the mu and delta receptors was the most potent and selective kappa opioid receptor antagonist identified.

Enhanced active contraction of the transversus abdominis during walking / Aumento de la contracción activa del músculo transverso del abdomen durante la caminata / Aumento da contração ativa do músculo transverso do abdome durante a caminhada

ABSTRACT Introduction: We applied three-dimensional gait analysis to assess the effects of enhanced active contraction of the transversus abdominis (EACTA) during walking. We sought to evaluate the effect of EACTA during walking in order to improve walking quality. Methods: Thirty college students were recruited and trained to perform EACTA during walking. We examined gait parameters under different conditions, including EACTA and habitual ACTA (natural walking with mild contraction of the feedforward mechanism of ACTA, HACTA) during walking using three-dimensional gait analysis. We compared differences in gait parameters under the two walking conditions using SPSS 16.0 statistical software. Results: The following gait parameters were significantly lower under EACTA conditions than under HACTA conditions (P < 0.05): stance phase, 59.151% ± 1.903% vs. 59.825% ± 1.495%; stride time, 1.104 s ± 0.080 s vs. 1.134 s ± 0.073 s:; stance time, 0.656 s ± 0.057 s vs. 0.678 s ± 0.053 s; and swing time, 0.447 s ± 0.028 s vs. 0.454 s ± 0.031 s, respectively. Gait parameters single support phase and mean velocity were significantly higher for EACTA than for HACTA conditions (both P < 0.05). Conclusions: Overall, the results revealed that EACTA during walking can improve gait. This method is simple, and EACTA training during walking to improve gait quality in daily life could provide a positive basis for people to strengthen the transverse abdominal muscle. Level of evidence III; Retrospective comparative study .

RESUMEN Introducción: Aplicamos el análisis tridimensional de la marcha para evaluar los efectos del aumento de la contracción activa del músculo transverso del abdomen (EACTA) durante la caminata. Buscamos evaluar el efecto del EACTA durante la caminata para mejorar su calidad. Métodos: Treinta estudiantes universitarios fueron reclutados y entrenados para realizar el EACTA durante la caminata. Examinamos los parámetros de la marcha en diferentes condiciones, incluyendo EACTA y ACTA habitual (caminata natural con leve contracción del mecanismo de feedforward del ACTA, HACTA) durante la caminata usando análisis tridimensional de la marcha. Comparamos las diferencias en los parámetros de la marcha en las dos condiciones de caminata en el software estadístico SPSS 16.0. Resultados: Los siguientes parámetros de marcha fueron significativamente más bajos en la condición EACTA que en condiciones HACTA (P <0,05): fase de apoyo 59,151 ± 1,903% vs 59,825 ± 1,495%, tiempo de zancada 1,104 s ± 0,080 s vs 1,134 s ± 0,073 s, tiempo de apoyo 0,656 s ± 0,057 s vs 0,678 s ± 0,053 s y tiempo de balance 0,447 s ± 0,028 s vs 0,454 s ± 0,031 s, respectivamente. Los parámetros de la marcha, fase de apoyo simple y velocidad promedio fueron significativamente mayores en el EACTA que en las condiciones HACTA (ambos P <0,05). Conclusiones: En general, los resultados revelaron que el EACTA durante la caminata puede mejorar la marcha. Este método es simple, y el entrenamiento del EACTA durante la caminata para mejorar la calidad de la marcha en la vida diaria puede ser una base positiva para el fortalecimiento del músculo transverso del abdomen. Nivel de evidencia III; Estudio retrospectivo comparativo .

RESUMO Introdução: Aplicamos a análise tridimensional da marcha para avaliar os efeitos do aumento da contração ativa do músculo transverso do abdome (EACTA) durante a caminhada. Procuramos avaliar o efeito do EACTA durante a caminhada para melhorar sua qualidade. Métodos: Trinta estudantes universitários foram recrutados e treinados para realizar o EACTA durante a caminhada. Examinamos os parâmetros da marcha em diferentes condições, incluindo EACTA e ACTA habitual (caminhada natural com leve contração do mecanismo de feedforward do ACTA, HACTA) durante a caminhada usando análise tridimensional da marcha. Comparamos as diferenças nos parâmetros da marcha nas duas condições de caminhada no software estatístico SPSS 16.0. Resultados: Os seguintes parâmetros da marcha foram significativamente mais baixos na condição EACTA do que em condições HACTA (P < 0,05): fase de apoio 59,151 ± 1,903% vs. 59,825 ± 1,495%, tempo de passada 1,104 s ± 0,080 s vs. 1,134 s ± 0,073 s, tempo de apoio 0,656 s ± 0,057 s vs. 0,678 s ± 0,053 s e tempo de balanço 0,447 s ± 0,028 s vs. 0,454 s ± 0,031 s, respectivamente. Os parâmetros da marcha fase de apoio simples e velocidade média foram significativamente maiores no EACTA do que nas condições HACTA (ambos P < 0,05). Conclusões: No geral, os resultados revelaram que o EACTA durante a caminhada pode melhorar a marcha. Esse método é simples, e o treinamento do EACTA durante a caminhada para melhorar a qualidade da marcha na vida diária pode ser uma base positiva para o fortalecimento do músculo transverso do abdome. Nível de evidência III; Estudo retrospectivo comparativo .

Modulation of nuclear factor-κB-mediated pro-inflammatory response is associated with exogenous administration of bone marrow-derived mesenchymal stem cells for treatment of experimental colitis.

Mesenchymal stem cells (MSCs) inhibit the immune response in vitro and prevent the induction of disease in certain experimental models. As a result, MSC­mediated therapy is a rapidly growing field of research. However, the efficacy of MSCs in the treatment of inflammatory bowel disease (IBD) has remained to be determined. In the present study, rats with 2,4,6­trinitrobenzene sulfonic acid (TNBS)­induced colitis were injected with prepared MSCs (1x106) into the tail vein. Two weeks following intravenous MSC administration, the concentration of tumor necrosis factor­α (TNF­α) in the serum was measured by an ELISA. The protein expression of nuclear factor­κB (NF­κBp65) in the colonic mucosa was assessed by western blot analysis. mRNA expression of TNF­α and NF­κBp65 was determined by reverse­transcription quantitative polymerase chain reaction. MSCs were shown to exert an immunomodulatory effect on TNBS­induced colitis and may be of use in the treatment of IBD. In addition, modulation of the NF­κB­mediated pro­inflammatory response may contribute to the underlying mechanism by which MSCs ameliorate the clinical and histological changes associated with IBD.

Extracellular Vesicles Derived from Bone Marrow Mesenchymal Stem Cells Protect against Experimental Colitis via Attenuating Colon Inflammation, Oxidative Stress and Apoptosis.

The administration of bone mesenchymal stem cells (BMSCs) could reverse experimental colitis, and the predominant mechanism in tissue repair seems to be related to their paracrine activity. BMSCs derived extracellular vesicles (BMSC-EVs), including mcirovesicles and exosomes, containing diverse proteins, mRNAs and micro-RNAs, mediating various biological functions, might be a main paracrine mechanism for stem cell to injured cell communication. We aimed to investigate the potential alleviating effects of BMSC-EVs in 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis model. Intravenous injection of BMSC-EVs attenuated the severity of colitis as evidenced by decrease of disease activity index (DAI) and histological colonic damage. In inflammation response, the BMSC-EVs treatment significantly reduced both the mRNA and protein levels of nuclear factor kappaBp65 (NF-κBp65), tumor necrosis factor-alpha (TNF-α), induciblenitric oxidesynthase (iNOS) and cyclooxygenase-2 (COX-2) in injured colon. Additionally, the BMSC-EVs injection resulted in a markedly decrease in interleukin-1ß (IL-1ß) and an increase in interleukin-10 (IL-10) expression. Therapeutic effect of BMSC-EVs associated with suppression of oxidative perturbations was manifested by a decrease in the activity of myeloperoxidase (MPO) and Malondialdehyde (MDA), as well as an increase in superoxide dismutase (SOD) and glutathione (GSH). BMSC-EVs also suppressed the apoptosis via reducing the cleavage of caspase-3, caspase-8 and caspase-9 in colitis rats. Data obtained indicated that the beneficial effects of BMSC-EVs were due to the down regulation of pro-inflammatory cytokines levels, inhibition of NF-κBp65 signal transduction pathways, modulation of anti-oxidant/ oxidant balance, and moderation of the occurrence of apoptosis.

Hydroxyurea analogues as kinetic and mechanistic probes of the nitric oxide producing reactions of hydroxyurea and oxyhemoglobin.

Derivatives of N-hydroxyurea that contain an N-hydroxy group react with oxyhemoglobin to form methemoglobin and variable amounts of nitrite/nitrate. Compounds with an unsubstituted -NHOH group produce the most nitrite/nitrate, which provides evidence for nitric oxide formation. The rate of reaction of these N-hydroxyurea derivatives with oxyhemoglobin correlates well with that compound's oxidation potential. Aromatic N-hydroxyureas react 25-80-fold faster with oxyhemoglobin than with N-hydroxyurea, suggesting other N-hydroxyurea analogues may be superior nitric oxide donors. Electron paramagnetic resonance spectroscopy shows that the formation of a low-spin methemoglobin-hydroxyurea complex is critical for iron nitrosyl hemoglobin formation. These results show that iron nitrosyl hemoglobin formation from the reaction of hydroxyureas and hemoglobin requires an unsubstituted -NHOH group and that the nitrogen atom of the non-N-hydroxy group must contain at least a single hydrogen atom. These results should guide the development of new hydroxyurea-based nitric oxide donors and sickle cell disease therapies.

Over-expression of CXCR4 on mesenchymal stem cells protect against experimental colitis via immunomodulatory functions in impaired tissue.

Bone marrow-derived mesenchymal stem cells (BMSCs) are attractive candidates for tissue regeneration and immunoregulation in inflammatory bowel disease. However, their in vivo reparative capability is limited owing to barren efficiency of BMSCs to injury region. Stromal cell-derived factor (SDF-1) plays an important role in chemotaxis and stem cell homing through interaction with its specific receptor CXC chemokine receptor 4 (CXCR4). The present study was designed to investigate the role of SDF-1α/CXCR4 axis in the therapeutic effects of lentivirus-preconditioned BMSCs for 2,4,6-trinitrobenzene sulfonic acid (TNBS)-colitis rats. BMSCs were isolated from female Sprague-Dawley rats and identified by flow cytometry. Lentiviral transduction was applied to over-express CXCR4/GFP (Ad-CXCR4-BMSCs) or null/GFP (Ad-GFP-BMSCs). Efficacy of engraftment was determined by the presence of enhanced green fluorescent protein (GFP) positive cells. One week after intravenous administration, Ad-GFP-BMSCs failed to colonize in the inflamed colon and had no beneficial effect in TNBS-induced colitis. Instead, Ad-CXCR4-BMSCs signally ameliorated both clinical and microanatomical severity of colitis. Immunofluorescence and western blotting showed that Ad-CXCR4-BMSCs migrated toward inflamed colon was more efficient than Ad-GFP-BMSCs. The therapeutic effect of Ad-CXCR4-BMSCs was mediated by the suppression of pro-inflammatory cytokines and STAT3 phosphorylation in injured colon. Collectively, our data indicated that over-expression CXCR4 led to enhance in vivo mobilization and engraftment of BMSCs into inflamed colon where these cells can function as an anti-inflammatory and immunomodulatory component of the immune system in TNBS-induced colitis.

Publication patterns and citation analysis of APJTM during 2008 and June 2014.

OBJECTIVE: To draw on data about publication patterns and citation indicators of Asian Pacific Journal of Tropical Medicine (APJTM) during 2008 and June 2014 in order to know about the current state of the journal. METHODS: Data used in this study were collected based on publications in APJTM from 2008 to June, 2014. Information including publication issue, type of manuscript, country/region of Corresponding author, funded research paper, and international collaboration were aggregated and analyzed with Excel software. Citation indicators including total cites, average cites of each manuscript, h-index, and impact factors were primarily drawn from Web of Science™ database on June 15, 2014 and changes over the past six and half years were interpreted. The top 10 most cited papers in Web of Science™ database were also analyzed. RESULTS: Number of all submissions has arisen from less than 200 in 2008 to over 1 500 in 2013, manuscript acceptance rate has decreased to be less than 14.00% indicating its improvement in quality over this period of time. Out of the 1 115 publiations, 23.77% were fruits of funded projects or produced by funded co-authors, 87.08% of all publications in APJTM were submited by authors from 10 most contributed countries. During the studied period, each published manuscript in the journal has received an average of 1.05 cites, and at least 10 publications has been cited for more 10 times. CONCLUSION: Detailed analysis shows APJTM has made great progress over the past six and half years, but authors' originating countries are still disproportionate. Efforts should be made to improve its citation indicators.

Study on the interactions between transplanted bone marrow-derived mesenchymal stem cells and regulatory T cells for the treatment of experimental colitis.

Accumulating evidence has established the use of mesenchymal stem cells (MSCs) as candidate cells for immunosuppressive therapy. Experimental studies have suggested that MSCs exert their immunomodulatory effects through the induction of regulatory T cells (Tregs) in vitro and in vivo. However, the interactions between MSCs and Tregs in inflammatory bowel disease (IBD) and whether MSCs can be used for the treatment of IBD remains to be elucidated. In this study, we aimed to investigate whether MSCs can be used for the treatment of IBD through the induction of Tregs. MSCs were isolated and identified by flow cytometry. The MSCs were transduced with a replication-defective recombinant lentiviral vector carrying GFP in order to be able to trace the injected cells in vivo. Prepared MSCs (1x106) were injected into rats with 2,4,6-trinitrobenzene sulfonic acid (TNBS)­induced colitis via the tail vein; the control rats received phosphate-buffered saline (PBS) alone. Two weeks after the intravenous infusion, the frequency of CD4+CD25+Foxp3 cells in the peripheral blood was examined by flow cytometry. The colon was sectioned and analyzed for histopathological changes. Foxp3 mRNA expression was determined by real-time reverse-transcription polymerase chain reaction (qRT-PCR). In our study, the systemic infusion of MSCs significantly ameliorated the clinical and histopathologic severity of TNBS-induced colitis in contrast to the controls. There was an inverse regulation of mucosal and peripheral Foxp3 expression, suggesting that the MSCs redistributed the Tregs from the mucosa to the blood. Thus, MSCs exhibit immunomodulatory functions and may be used to ameliorate or treat IBD by redistributing regulatory T cells. Therefore, the interactions between transplanted bone marrow-derived MSCs and Tregs should be further investigated; MSCs have tremendous potential for use in the treatment of IBD.