Skeletal muscle secretome in Duchenne muscular dystrophy: a pivotal anti-inflammatory role of adiponectin

Background

Persistent inflammation exacerbates the progression of Duchenne muscular dystrophy (DMD). The hormone, adiponectin (ApN), which is decreased in the metabolic syndrome, exhibits anti-inflammatory properties on skeletal muscle and alleviates the dystrophic phenotype of mdx mice. Here, we investigate whether ApN retains its anti-inflammatory action in myotubes obtained from DMD patients. We unravel the underlying mechanisms by studying the secretome and the early events of ApN.

Methods

Primary cultures of myotubes from DMD and control patients were treated or not by ApN after an inflammatory challenge. Myokines secreted in medium were identified by cytokine antibody-arrays and ELISAs. The early events of ApN signaling were assessed by abrogating selected genes.

Results

ApN retained its anti-inflammatory properties in both dystrophic and control myotubes. Profiling of secretory products revealed that ApN downregulated the secretion of two pro-inflammatory factors (TNFα and IL-17A), one soluble receptor (sTNFRII), and one chemokine (CCL28) in DMD myotubes, while upregulating IL-6 that exerts some anti-inflammatory effects. These changes were explained by pretranslational mechanisms. Earlier events of the ApN cascade involved AdipoR1, the main receptor for muscle, and the AMPK-SIRT1-PGC-1α axis leading, besides alteration of the myokine profile, to the upregulation of utrophin A (a dystrophin analog).

Conclusion

ApN retains its beneficial properties in dystrophic muscles by activating the AdipoR1-AMPK-SIRT1-PGC-1α pathway, thereby inducing a shift in the secretion of downstream myokines toward a less inflammatory profile while upregulating utrophin. ApN, the early events of the cascade and downstream myokines may be therapeutic targets for the management of DMD.

     

Eukaryotic DNA damage checkpoint activation in response to double-strand breaks

Double-strand breaks (DSBs) are the most detrimental form of DNA damage. Failure to repair these cytotoxic lesions can result in genome rearrangements conducive to the development of many diseases, including cancer. The DNA damage response (DDR) ensures the rapid detection and repair of DSBs in order to maintain genome integrity. Central to the DDR are the DNA damage checkpoints. When activated by DNA damage, these sophisticated surveillance mechanisms induce transient cell cycle arrests, allowing sufficient time for DNA repair. Since the term “checkpoint” was coined over 20 years ago, our understanding of the molecular mechanisms governing the DNA damage checkpoint has advanced significantly. These pathways are highly conserved from yeast to humans. Thus, significant findings in yeast may be extrapolated to vertebrates, greatly facilitating the molecular dissection of these complex regulatory networks. This review focuses on the cellular response to DSBs in Saccharomyces cerevisiae, providing a comprehensive overview of how these signalling pathways function to orchestrate the cellular response to DNA damage and preserve genome stability in eukaryotic cells.     

Reflection-in-Action: Modeling a specific organization through the Viable Systems Model

The purpose of this project is to explore a way for systems practitioners to learn the skills necessary to employ a particular systems model in practice. Reflection-in-Action is one way to embark on a double-loop learning cycle and, thus, develop the tacit skills and inarticulate knowledge needed to employ a particular systems model. Through Reflection-in-Action, a Viable Systems Model (VSM) of the San Francisco Zoo was constructed. This project involved three phases of research; for each, a VSM using the data available was constructed. During each phase, the questioning of current assumptions and operating norms led to changes in the direction of the research. This led to the construction of significantly different VSMs for each phase.     

A population of human lymphocytes staining for esterases

Summary A population of lymphocytes is found to stain positively for esterases. The positively staining lymphocytes are more predominant among T than B lymphocytes and are significantly increased on stimulation with PHA. Treatment with cholinestrase inhibitors reduces their number significantly.This work was supported by a grant from the Medical Research Council of Canada.     

Deep resequencing of GWAS loci identifies independent rare variants associated with inflammatory bowel disease

More than 1,000 susceptibility loci have been identified through genome-wide association studies (GWAS) of common variants; however, the specific genes and full allelic spectrum of causal variants underlying these findings have not yet been defined. Here we used pooled next-generation sequencing to study 56 genes from regions associated with Crohn's disease in 350 cases and 350 controls. Through follow-up genotyping of 70 rare and low-frequency protein-altering variants in nine independent case-control series (16,054 Crohn's disease cases, 12,153 ulcerative colitis cases and 17,575 healthy controls), we identified four additional independent risk factors in NOD2, two additional protective variants in IL23R, a highly significant association with a protective splice variant in CARD9 (P < 1 × 10(-16), odds ratio ≈ 0.29) and additional associations with coding variants in IL18RAP, CUL2, C1orf106, PTPN22 and MUC19. We extend the results of successful GWAS by identifying new, rare and probably functional variants that could aid functional experiments and predictive models.     

Resculpting the binding pocket of APC superfamily LeuT-fold amino acid transporters

Amino acid transporters are essential components of prokaryote and eukaryote cells, possess distinct physiological functions, and differ markedly in substrate specificity. Amino acid transporters can be both drug targets and drug transporters (bioavailability, targeting) with many monogenic disorders resulting from dysfunctional membrane transport. The largest collection of amino acid transporters (including the mammalian SLC6, SLC7, SLC32, SLC36, and SLC38 families), across all kingdoms of life, is within the Amino acid-Polyamine-organoCation (APC) superfamily. The LeuT-fold is a paradigm structure for APC superfamily amino acid transporters and carriers of sugars, neurotransmitters, electrolytes, osmolytes, vitamins, micronutrients, signalling molecules, and organic and fatty acids. Each transporter is specific for a unique sub-set of solutes, specificity being determined by how well a substrate fits into each binding pocket. However, the molecular basis of substrate selectivity remains, by and large, elusive. Using an integrated computational and experimental approach, we demonstrate that a single position within the LeuT-fold can play a crucial role in determining substrate specificity in mammalian and arthropod amino acid transporters within the APC superfamily. Systematic mutation of the amino acid residue occupying the equivalent position to LeuT V104 titrates binding pocket space resulting in dramatic changes in substrate selectivity in exemplar APC amino acid transporters including PAT2 (SLC36A2) and SNAT5 (SLC38A5). Our work demonstrates how a single residue/site within an archetypal structural motif can alter substrate affinity and selectivity within this important superfamily of diverse membrane transporters.     

IDEA: A Collaborative Organizational Design Process Integrating Innovation, Design, Engagement, and Action

This paper describes an innovative and successful 1-year organization change process. It captures a design-based inquiry that simultaneously applies creative, purposeful, and systemic thinking to a complex set of issues. Three significant findings result from this research. First, this paper discusses how the change process created the necessary and sufficient conditions allowing for the creation of an innovative organizational design that embeds both optimization and innovation. Second, Design Thinking was used to develop a 2-day participative design process we have called IDEA, an acronym for integrating innovation, design, engagement, and action. We believe that the IDEA organizational design process is replicable. Third, it describes an emergent and co-created change process. This paper concludes by raising questions for future transformative organizational design efforts.     

热输入对深海X70粗晶区组织和韧性的影响

为解决深海X70管线钢在实际焊接中粗晶区(CGHAZ)的脆化问题,在不同热循环工艺下对X70管线钢进行了热模拟研究。采用Gleeble-3800热模拟机模拟X70管线钢CGHAZ,研究CGHAZ在10～60 kJ/cm不同热输入(HI)条件下组织和韧性的变化规律,并通过光学显微镜(OM)、扫描电镜(SEM)和夏比冲击试验等手段表征CGHAZ的组织和韧性。结果表明,不同热输入下试验钢的组织主要由粒状贝氏体(GB)、贝氏体铁素体(BF)和马-奥组元(M-A组元)组成;当HI不断增大时,BF比例减少,GB比例增加,M-A组元粗化,冲击吸收能先升高再降低;当HI为20 kJ/cm时,BF和GB可获得优异组合,断口为韧性断裂,冲击吸收能达到173.8 J;当HI大于20 kJ/cm时,断口解离断裂,冲击吸收能下降明显,最低为18.8 J。因此,较低的热输入可提高CGHAZ的韧性,使X70管线钢具有高强度、高韧性和良好的焊接性。研究结果可为优化焊接工艺提供理论依据。