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Sample records for flag-tagged micro dystrophin

  1. TNF-α-Induced microRNAs Control Dystrophin Expression in Becker Muscular Dystrophy.

    Science.gov (United States)

    Fiorillo, Alyson A; Heier, Christopher R; Novak, James S; Tully, Christopher B; Brown, Kristy J; Uaesoontrachoon, Kitipong; Vila, Maria C; Ngheim, Peter P; Bello, Luca; Kornegay, Joe N; Angelini, Corrado; Partridge, Terence A; Nagaraju, Kanneboyina; Hoffman, Eric P

    2015-09-08

    The amount and distribution of dystrophin protein in myofibers and muscle is highly variable in Becker muscular dystrophy and in exon-skipping trials for Duchenne muscular dystrophy. Here, we investigate a molecular basis for this variability. In muscle from Becker patients sharing the same exon 45-47 in-frame deletion, dystrophin levels negatively correlate with microRNAs predicted to target dystrophin. Seven microRNAs inhibit dystrophin expression in vitro, and three are validated in vivo (miR-146b/miR-374a/miR-31). microRNAs are expressed in dystrophic myofibers and increase with age and disease severity. In exon-skipping-treated mdx mice, microRNAs are significantly higher in muscles with low dystrophin rescue. TNF-α increases microRNA levels in vitro whereas NFκB inhibition blocks this in vitro and in vivo. Collectively, these data show that microRNAs contribute to variable dystrophin levels in muscular dystrophy. Our findings suggest a model where chronic inflammation in distinct microenvironments induces pathological microRNAs, initiating a self-sustaining feedback loop that exacerbates disease progression. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  2. TNF-α-Induced microRNAs Control Dystrophin Expression in Becker Muscular Dystrophy

    Directory of Open Access Journals (Sweden)

    Alyson A. Fiorillo

    2015-09-01

    Full Text Available The amount and distribution of dystrophin protein in myofibers and muscle is highly variable in Becker muscular dystrophy and in exon-skipping trials for Duchenne muscular dystrophy. Here, we investigate a molecular basis for this variability. In muscle from Becker patients sharing the same exon 45–47 in-frame deletion, dystrophin levels negatively correlate with microRNAs predicted to target dystrophin. Seven microRNAs inhibit dystrophin expression in vitro, and three are validated in vivo (miR-146b/miR-374a/miR-31. microRNAs are expressed in dystrophic myofibers and increase with age and disease severity. In exon-skipping-treated mdx mice, microRNAs are significantly higher in muscles with low dystrophin rescue. TNF-α increases microRNA levels in vitro whereas NFκB inhibition blocks this in vitro and in vivo. Collectively, these data show that microRNAs contribute to variable dystrophin levels in muscular dystrophy. Our findings suggest a model where chronic inflammation in distinct microenvironments induces pathological microRNAs, initiating a self-sustaining feedback loop that exacerbates disease progression.

  3. mRNA and microRNA transcriptomics analyses in a murine model of dystrophin loss and therapeutic restoration

    Directory of Open Access Journals (Sweden)

    Thomas C. Roberts

    2016-03-01

    Full Text Available Duchenne muscular dystrophy (DMD is a pediatric, X-linked, progressive muscle-wasting disorder caused by loss of function mutations affecting the gene encoding the dystrophin protein. While the primary genetic insult in DMD is well described, many details of the molecular and cellular pathologies that follow dystrophin loss are incompletely understood. To investigate gene expression in dystrophic muscle we have applied mRNA and microRNA (miRNA microarray technology to the mdx mouse model of DMD. This study was designed to generate a complete description of gene expression changes associated with dystrophic pathology and the response to an experimental therapy which restores dystrophin protein function. These datasets have enabled (1 the determination of gene expression changes associated with dystrophic pathology, (2 identification of differentially expressed genes that are restored towards wild-type levels after therapeutic dystrophin rescue, (3 investigation of the correlation between mRNA and protein expression (determined by parallel mass spectrometry proteomics analysis, and (4 prediction of pathology associated miRNA-target interactions. Here we describe in detail how the data were generated including the basic analysis as contained in the manuscript published in Human Molecular Genetics with PMID 26385637. The data have been deposited in the Gene Expression Omnibus (GEO with the accession number GSE64420.

  4. Bioenergetic Consequences of FLAG Tag Addition to the C-Terminus of Subunit 8 of Yeast Saccharomyces cerevisiae Mitochondrial ATP Synthase

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    I MADE ARTIKA

    2010-09-01

    Full Text Available The yeast mitochondrial F1F0-ATP synthase is a multisubunit complex that contains at least 17 different subunits. Subunit 8 of yeast mitochondrial ATP synthase is a hydrophobic protein of 48 amino acids encoded by the mitochondrial ATP8 gene. Subunit 8 has three distinct domains; an N-terminal domain, a central hydrophobic domain and a C-terminal domain. FLAG tag addition to subunit 8 protein potentially facilitate elucidation of its topology, structure, and function. It has been shown that following incorporation of FLAG tag to its C-terminus, subunit 8 still assemble into functional ATP synthase complex. In order to analyze bioenergetic consequences of the FLAG tag addition, a yeast strain expressing FLAG tagged-subunit 8 was subjected to cellular respiration assays. Results obtained showed that addition of FLAG tag to the C-terminus of subunit 8 does not impair its proper functioning. The FLAG tag system, therefore, can be employed to study subunit 8′s detailed structure, topology, and function.

  5. Hepatitis C virus expressing flag-tagged envelope protein 2 has unaltered infectivity and density, is specifically neutralized by flag antibodies and can be purified by affinity chromatography

    DEFF Research Database (Denmark)

    Prentø, Jannick Cornelius; Bukh, Jens

    2011-01-01

    to the original virus. Flag-tagged virus was susceptible to flag-specific antibody neutralization, and infected cells could be immuno-stained by anti-flag antibodies. Using affinity chromatography with anti-flag resin we repeatedly obtained ~30% recovery of infectious particles. The full viability and unaltered...

  6. A Translational Pathway Toward a Clinical Trial Using the Second-Generation AAV Micro-Dystrophin Vector

    Science.gov (United States)

    2016-09-01

    mune system a few weeks later. It is now clear that the gene delivery vehicle (AAV virus capsid), cargo (transgene), or the protein produced from the...Ideally, delivery of a full-length dystrophin cDNA will yield the production of a full- length dystrophin protein and the maximum pro- tection of...investigational new drug (IND) application can be filed for a gene therapy trial with systemic delivery of dystrophin? Dr. Duan: A number of IND

  7. A translational approach for limb vascular delivery of the micro-dystrophin gene without high volume or high pressure for treatment of Duchenne muscular dystrophy

    Directory of Open Access Journals (Sweden)

    Chicoine Louis G

    2007-09-01

    Full Text Available Abstract Background Duchenne muscular dystrophy (DMD is an X-linked recessive disorder with monogenic mutations setting the stage for successful gene therapy treatment. We have completed a study that directly deals with the following key issues that can be directly adapted to a gene therapy clinical trial using rAAV considering the following criteria: 1 A regional vascular delivery approach that will protect the patient from widespread dissemination of virus; 2 an approach to potentially facilitate safe passage of the virus for efficient skeletal muscle transduction; 3 the use of viral doses to accommodate current limitations imposed by vector production methods; 4 and at the same time, achieve a clinically meaningful outcome by transducing multiple muscles in the lower limb to prolong ambulation. Methods The capacity of AAV1, AAV6 or AAV8 to cross the vascular endothelial barrier carrying a micro-dystrophin cDNA was compared under identical conditions with delivery through a catheter placed in the femoral artery of the mdx mouse. Transduction efficiency was assessed by immuno-staining using an antibody (Manex1a that recognizes the N-terminus of micro-dystrophin. The degree of physiologic correction was assessed by measuring tetanic force and protection from eccentric contraction in the extensor digitorum longus muscle (EDL. The vascular delivery paradigm found successful in the mouse was carried to the non-human primate to test its potential translation to boys with DMD. Results Regional vascular delivery resulted in transduction by rAAV8.micro-dystrophin reaching 94.5 ± 0.9 (1 month, 91.3 ± 3.1 (2 months, and 89.6 ± 1.6% (3 months. rAAV6.micro-dystrophin treated animals demonstrated 87.7 ± 6.8 (1 month, 78.9 ± 7.4 (2 months, and 81.2 ± 6.2% (3 months transduction. In striking contrast, rAAV1 demonstrated very low transduction efficiency [0.9 ± 0.3 (1 month, 2.1 ± 0.8 (2 months, and 2.1 ± 0.7% (3 months] by vascular delivery. Micro-dystrophin

  8. FLAG-tagged CD19-specific CAR-T cells eliminate CD19-bearing solid tumor cells in vitro and in vivo.

    Science.gov (United States)

    Berahovich, Robert; Xu, Shirley; Zhou, Hua; Harto, Hizkia; Xu, Qumiao; Garcia, Andres; Liu, Fenyong; Golubovskaya, Vita M; Wu, Lijun

    2017-06-01

    Autologous T cells expressing chimeric antigen receptors (CARs) specific for CD19 have demonstrated remarkable efficacy as therapeutics for B cell malignancies. In the present study, we generated FLAG-tagged CD19-specific CAR-T cells (CD19-FLAG) and compared them to their non-tagged counterparts for their effects on solid and hematological cancer cells in vitro and in vivo . For solid tumors, we used HeLa cervical carcinoma cells engineered to overexpress CD19 (HeLa-CD19), and for hematological cancer we used Raji Burkitt's lymphoma cells, which endogenously express CD19. Like non-tagged CD19 CAR-T cells, CD19-FLAG CAR-T cells expanded in culture >100-fold and exhibited potent cytolytic activity against both HeLa-CD19 and Raji cells in vitro . CD19-FLAG CAR-T cells also secreted significantly more IFN-gamma and IL-2 than the control T cells. In vivo , CD19-FLAG CAR-T cells significantly blocked the growth of HeLa-CD19 solid tumors, increased tumor cleaved caspase-3 levels, and expanded systemically. CD19-FLAG CAR-T cells also significantly reduced Raji tumor burden and extended mouse survival. These results demonstrate the strong efficacy of FLAG-tagged CD19 CAR-T cells in solid and hematological cancer models.

  9. Utrophin Compensates dystrophin Loss during Mouse Spermatogenesis

    OpenAIRE

    Chen, Hung-Chih; Chin, Yu-Feng; Lundy, David J.; Liang, Chung-Tiang; Chi, Ya-Hui; Kuo, Paolin; Hsieh, Patrick C. H.

    2017-01-01

    Duchenne muscular dystrophy (DMD) is an X-linked genetic disorder resulting from mutations in the dystrophin gene. The mdx/utrn ?/? mouse, lacking in both dystrophin and its autosomal homologue utrophin, is commonly used to model the clinical symptoms of DMD. Interestingly, these mice are infertile but the mechanisms underlying this phenomenon remain unclear. Using dystrophin deficient mdx mouse and utrophin haplodeficient mdx/utrn +/? mouse models, we demonstrate the contribution of Dp427 (f...

  10. Dystrophin Immunity in Duchenne’s Muscular Dystrophy

    OpenAIRE

    Mendell, Jerry R.; Campbell, Katherine; Rodino-Klapac, Louise; Sahenk, Zarife; Shilling, Chris; Lewis, Sarah; Bowles, Dawn; Gray, Steven; Li, Chengwen; Galloway, Gloria; Malik, Vinod; Coley, Brian; Clark, K. Reed; Li, Juan; Xiao, Xiao

    2010-01-01

    We report on delivery of a functional dystrophin transgene to skeletal muscle in six patients with Duchenne’s muscular dystrophy. Dystrophin-specific T cells were detected after treatment, providing evidence of transgene expression even when the functional protein was not visualized in skeletal muscle. Circulating dystrophin-specific T cells were unexpectedly detected in two patients before vector treatment. Revertant dystrophin fibers, which expressed functional, truncated dystrophin from th...

  11. Functional substitution by TAT-utrophin in dystrophin-deficient mice.

    Directory of Open Access Journals (Sweden)

    Kevin J Sonnemann

    2009-05-01

    Full Text Available The loss of dystrophin compromises muscle cell membrane stability and causes Duchenne muscular dystrophy and/or various forms of cardiomyopathy. Increased expression of the dystrophin homolog utrophin by gene delivery or pharmacologic up-regulation has been demonstrated to restore membrane integrity and improve the phenotype in the dystrophin-deficient mdx mouse. However, the lack of a viable therapy in humans predicates the need to explore alternative methods to combat dystrophin deficiency. We investigated whether systemic administration of recombinant full-length utrophin (Utr or DeltaR4-21 "micro" utrophin (muUtr protein modified with the cell-penetrating TAT protein transduction domain could attenuate the phenotype of mdx mice.Recombinant TAT-Utr and TAT-muUtr proteins were expressed using the baculovirus system and purified using FLAG-affinity chromatography. Age-matched mdx mice received six twice-weekly intraperitoneal injections of either recombinant protein or PBS. Three days after the final injection, mice were analyzed for several phenotypic parameters of dystrophin deficiency. Injected TAT-muUtr transduced all tissues examined, integrated with members of the dystrophin complex, reduced serum levels of creatine kinase (11,290+/-920 U versus 5,950+/-1,120 U; PBS versus TAT, the prevalence of muscle degeneration/regeneration (54%+/-5% versus 37%+/-4% of centrally nucleated fibers; PBS versus TAT, the susceptibility to eccentric contraction-induced force drop (72%+/-5% versus 40%+/-8% drop; PBS versus TAT, and increased specific force production (9.7+/-1.1 N/cm(2 versus 12.8+/-0.9 N/cm(2; PBS versus TAT.These results are, to our knowledge, the first to establish the efficacy and feasibility of TAT-utrophin-based constructs as a novel direct protein-replacement therapy for the treatment of skeletal and cardiac muscle diseases caused by loss of dystrophin.

  12. Dystrophin analysis in carriers of Duchenne and Becker muscular dystrophy

    NARCIS (Netherlands)

    Hoogerwaard, Edo M.; Ginjaar, Ieke B.; Bakker, Egbert; de Visser, Marianne

    2005-01-01

    Associations between clinical phenotype (muscle weakness, dilated cardiomyopathy) and dystrophin abnormalities in muscle tissue among definite carriers of Duchenne (DMD) and Becker muscular dystrophy (BMD) were investigated. No associations between dystrophin abnormalities and clinical variables in

  13. Dystrophin Immunity in Duchenne’s Muscular Dystrophy

    Science.gov (United States)

    Mendell, Jerry R.; Campbell, Katherine; Rodino-Klapac, Louise; Sahenk, Zarife; Shilling, Chris; Lewis, Sarah; Bowles, Dawn; Gray, Steven; Li, Chengwen; Galloway, Gloria; Malik, Vinod; Coley, Brian; Clark, K. Reed; Li, Juan; Xiao, Xiao; Samulski, Jade; McPhee, Scott W.; Samulski, R. Jude; Walker, Christopher M.

    2010-01-01

    SUMMARY We report on delivery of a functional dystrophin transgene to skeletal muscle in six patients with Duchenne’s muscular dystrophy. Dystrophin-specific T cells were detected after treatment, providing evidence of transgene expression even when the functional protein was not visualized in skeletal muscle. Circulating dystrophin-specific T cells were unexpectedly detected in two patients before vector treatment. Revertant dystrophin fibers, which expressed functional, truncated dystrophin from the deleted endogenous gene after spontaneous in-frame splicing, contained epitopes targeted by the autoreactive T cells. The potential for T-cell immunity to self and nonself dystrophin epitopes should be considered in designing and monitoring experimental therapies for this disease. (Funded by the Muscular Dystrophy Association and others; ClinicalTrials.gov number, NCT00428935.) PMID:20925545

  14. Laryngeal Muscles Are Spared in the Dystrophin Deficient "mdx" Mouse

    Science.gov (United States)

    Thomas, Lisa B.; Joseph, Gayle L.; Adkins, Tracey D.; Andrade, Francisco H.; Stemple, Joseph C.

    2008-01-01

    Purpose: "Duchenne muscular dystrophy (DMD)" is caused by the loss of the cytoskeletal protein, dystrophin. The disease leads to severe and progressive skeletal muscle wasting. Interestingly, the disease spares some muscles. The purpose of the study was to determine the effects of dystrophin deficiency on 2 intrinsic laryngeal muscles, the…

  15. Evaluation of point mutations in dystrophin gene in Iranian ...

    Indian Academy of Sciences (India)

    5Department of Biology, Science and Research Branch, Islamic Azad ... Dystrophin protein is found ... Duchenne and Becker muscular dystrophy; neuromuscular disorder; point mutation. ..... modern diagnostic techniques to a large cohort.

  16. Marginal level dystrophin expression improves clinical outcome in a strain of dystrophin/utrophin double knockout mice.

    Directory of Open Access Journals (Sweden)

    Dejia Li

    2010-12-01

    Full Text Available Inactivation of all utrophin isoforms in dystrophin-deficient mdx mice results in a strain of utrophin knockout mdx (uko/mdx mice. Uko/mdx mice display severe clinical symptoms and die prematurely as in Duchenne muscular dystrophy (DMD patients. Here we tested the hypothesis that marginal level dystrophin expression may improve the clinical outcome of uko/mdx mice. It is well established that mdx3cv (3cv mice express a near-full length dystrophin protein at ∼5% of the normal level. We crossed utrophin-null mutation to the 3cv background. The resulting uko/3cv mice expressed the same level of dystrophin as 3cv mice but utrophin expression was completely eliminated. Surprisingly, uko/3cv mice showed a much milder phenotype. Compared to uko/mdx mice, uko/3cv mice had significantly higher body weight and stronger specific muscle force. Most importantly, uko/3cv outlived uko/mdx mice by several folds. Our results suggest that a threshold level dystrophin expression may provide vital clinical support in a severely affected DMD mouse model. This finding may hold clinical implications in developing novel DMD therapies.

  17. Proteomic analysis reveals new cardiac-specific dystrophin-associated proteins.

    Directory of Open Access Journals (Sweden)

    Eric K Johnson

    Full Text Available Mutations affecting the expression of dystrophin result in progressive loss of skeletal muscle function and cardiomyopathy leading to early mortality. Interestingly, clinical studies revealed no correlation in disease severity or age of onset between cardiac and skeletal muscles, suggesting that dystrophin may play overlapping yet different roles in these two striated muscles. Since dystrophin serves as a structural and signaling scaffold, functional differences likely arise from tissue-specific protein interactions. To test this, we optimized a proteomics-based approach to purify, identify and compare the interactome of dystrophin between cardiac and skeletal muscles from as little as 50 mg of starting material. We found selective tissue-specific differences in the protein associations of cardiac and skeletal muscle full length dystrophin to syntrophins and dystrobrevins that couple dystrophin to signaling pathways. Importantly, we identified novel cardiac-specific interactions of dystrophin with proteins known to regulate cardiac contraction and to be involved in cardiac disease. Our approach overcomes a major challenge in the muscular dystrophy field of rapidly and consistently identifying bona fide dystrophin-interacting proteins in tissues. In addition, our findings support the existence of cardiac-specific functions of dystrophin and may guide studies into early triggers of cardiac disease in Duchenne and Becker muscular dystrophies.

  18. Evolutionary study of vertebrate and invertebrate members of the dystrophin and utrophin gene family

    Energy Technology Data Exchange (ETDEWEB)

    Roberts, R.G.; Nicholson, L.; Bobrow, M. [Paediatric Research Unit, London (United Kingdom)] [and others

    1994-09-01

    Vertebrates express two members of the dystrophin gene family. The prototype, dystrophin, is expressed in muscle and neural tissue, and is defective in the human disorders Duchenne and Becker muscular dystrophy (DMD, BMD). The dystrophin homologue utrophin is more generally expressed but has not yet been associated with a genetic disorder. The function of neither protein is clear. A comparison of human utrophin with the known dystrophins (human, mouse, chicken, Torpedo) suggests that dystrophin and utrophin diverged before the vertebrate radiation. We have used reverse-transcript PCR (RT-PCR) directed by degenerate primers to characterize dystrophin and utrophin transcripts from a range of vertebrate and invertebrate animals. Our results suggest that the duplication leading to distinct dystrophin and utrophin genes occurred close to the point of divergence of urochordates from the cephalochordate-vertebrate lineage. This divergence may have occurred to fulfill a novel role which arose at this point, or may reflect a need for separate regulation of the neuromuscular and other functions of the ancient dystrophin. Our data include sequences of the first non-human utrophins to be characterized, and show these to be substantially more divergent than their cognate dystrophins. In addition, our results provide a large body of information regarding the tolerance of amino acid positions in the cysteine-rich and C-terminal domains to substitution. This will aid the interpretations of DMD and BMD missense mutations in these regions.

  19. Dystrophin in frameshift deletion patients with Becker Muscular Dystrophy

    Energy Technology Data Exchange (ETDEWEB)

    Gangopadhyay, S.B.; Ray, P.N.; Worton, R.G.; Sherratt, T.G.; Heckmatt, J.Z.; Dubowitz, V.; Strong, P.N.; Miller, G. (Penn State College of Medicine, Hershey, PA (United States)); Shokeir, M. (Univ. Hospital, Saskatchewan (Canada))

    1992-09-01

    In a previous study the authors identified 14 cases with Duchenne muscular dystrophy (DMD) or its milder variant, Becker muscular dystrophy (BMD), with a deletion of exons 3-7, a deletion that would be expected to shift the translational reading frame of the mRNA and give a severe phenotype. They have examined dystrophin and its mRNA from muscle biopsies of seven cases with either mild or intermediate phenotypes. In all cases they detected slightly lower-molecular-weight dystrophin in 12%-15% abundance relative to the normal. By sequencing amplified mRNA they have found that exon 2 is spliced to exon 8, a splice that produces a frameshifted mRNA, and have found no evidence for alternate splicing that might be involved in restoration of dystrophin mRNA reading frame in the patients with a mild phenotype. Other transcriptional and posttranscriptional mechanisms such as cryptic promoter, ribosomal frameshifting, and reinitiation are suggested that might play some role in restoring the reading frame. 34 refs., 5 figs. 1 tab.

  20. Spectrum of small mutations in the dystrophin coding region

    Energy Technology Data Exchange (ETDEWEB)

    Prior, T.W.; Bartolo, C.; Pearl, D.K. [Ohio State Univ., Columbus, OH (United States)] [and others

    1995-07-01

    Duchenne and Becker muscular dystrophies (DMD and BMD) are caused by defects in the dystrophin gene. About two-thirds of the affected patients have large deletions or duplications, which occur in the 5` and central portion of the gene. The nondeletion/duplication cases are most likely the result of smaller mutations that cannot be identified by current diagnostic screening strategies. We screened {approximately} 80% of the dystrophin coding sequence for small mutations in 158 patients without deletions or duplications and identified 29 mutations. The study indicates that many of the DMD and the majority of the BMD small mutations lie in noncoding regions of the gene. All of the mutations identified were unique to single patients, and most of the mutations resulted in protein truncation. We did not find a clustering of small mutations similar to the deletion distribution but found > 40% of the small mutations 3` of exon 55. The extent of protein truncation caused by the 3` mutations did not determine the phenotype, since even the exon 76 nonsense mutation resulted in the severe DMD phenotype. Our study confirms that the dystrophin gene is subject to a high rate of mutation in CpG sequences. As a consequence of not finding any hotspots or prevalent small mutations, we conclude that it is presently not possible to perform direct carrier and prenatal diagnostics for many families without deletions or duplications. 71 refs., 2 figs., 2 tabs.

  1. Increased susceptibility of dystrophin-deficient brain to mild hypoxia

    International Nuclear Information System (INIS)

    Wallis, T.; Rae, C.; Bubb, W.A.; Head, S.I.

    2002-01-01

    Full text: Duchenne muscular dystrophy is an X-linked disorder resulting from total absence of the 427 kDa protein dystrophin. Dystrophin is normally expressed in the brain mainly in a neuronal subpopulation: cortical pyramidal cells, hippocampal CA1 neurons and cerebellar Purkinje cells. One suggested role for dystrophin is in colocalising mitochondrial creatine kinase with ADP translocase and ATP synthase in mitochondria. Brain tissue slices in the murine model of Duchenne dystrophy, the mdx mouse, have been shown to be more sensitive to hypoxia than control. In this work, we used 13 C NMR to monitor the metabolic response of mdx cortical brain tissue slices to normoxia (95%O 2 /5% CO 2 ) and mild hypoxia (95%air/5% CO 2 ). Under normoxic conditions, mdx cortical slices displayed increased net flux through the Krebs cycle and glutamate/glutamine cycle, consistent with the proposed GABA A lesion which results in decreased inhibitory input. By contrast, mild hypoxia resulted in a significant increase in the total pool size of lactate and decreased net flux of 13 C from [3- 13 C]pyruvate into glutamate C4, GABA C2 and Ala C2, as well as decreased anaplerotic activity as measured by the ratio of Asp C2: Asp C3 label. Mild hypoxia has a significantly greater effect on brain oxidative metabolism in mdx mice, than in control

  2. Targeted Exon Skipping to Address “Leaky” Mutations in the Dystrophin Gene

    Directory of Open Access Journals (Sweden)

    Sue Fletcher

    2012-01-01

    Full Text Available Protein-truncating mutations in the dystrophin gene lead to the progressive muscle wasting disorder Duchenne muscular dystrophy, whereas in-frame deletions typically manifest as the milder allelic condition, Becker muscular dystrophy. Antisense oligomer-induced exon skipping can modify dystrophin gene expression so that a disease-associated dystrophin pre-mRNA is processed into a Becker muscular dystrophy-like mature transcript. Despite genomic deletions that may encompass hundreds of kilobases of the gene, some dystrophin mutations appear “leaky”, and low levels of high molecular weight, and presumably semi-functional, dystrophin are produced. A likely causative mechanism is endogenous exon skipping, and Duchenne individuals with higher baseline levels of dystrophin may respond more efficiently to the administration of splice-switching antisense oligomers. We optimized excision of exons 8 and 9 in normal human myoblasts, and evaluated several oligomers in cells from eight Duchenne muscular dystrophy patients with deletions in a known “leaky” region of the dystrophin gene. Inter-patient variation in response to antisense oligomer induced skipping in vitro appeared minimal. We describe oligomers targeting exon 8, that unequivocally increase dystrophin above baseline in vitro, and propose that patients with leaky mutations are ideally suited for participation in antisense oligomer mediated splice-switching clinical studies.

  3. The influence of low dystrophin levels on disease pathology in mouse models for Duchenne Muscular Dystrophy

    NARCIS (Netherlands)

    Putten, Maaike van

    2013-01-01

    Duchenne muscular dystrophy (DMD) is the most prevalent neuromuscular disorder, caused by mutations in the DMD gene that prevent synthesis of dystrophin. Fibers that lack dystrophin are sensitive to exercise-induced damage, resulting in progressive muscle wasting, loss of ambulation and premature

  4. Carrier detection of duchenne and becker muscular dystrophy using muscle dystrophin immunohistochemistry

    Directory of Open Access Journals (Sweden)

    Acary S. Bulle Oliveira

    1992-12-01

    Full Text Available To ascertain whether dystrophin immunohistochemistry could improve DMD/ BMD carrier detection, we analyzed 14 muscle biopsies from 13 DMD and one BMD probable and possible carriers. All women were also evaluated using conventional methods, including genetic analysis, clinical and neurological evaluation, serum CK levels, KMG, and muscle biopsy. In 6 cases, there was a mosaic of dystrophin-positive and dystrophin-deficient fibers that allowed to make the diagnosis of a carrier state. Comparing dystrophin immunohistochemistry to the traditional methods, it was noted that this method is less sensitive than serum CK measuremens, but is more sensitive than EMG and muscle biopsy. The use of dystrophin immunohistochemistry in addition to CK, EMG and muscle biopsy improved the accuracy of carrier detection. This method is also helpful to distinguish manifesting DMD carriers from patients with other neuromuscular diseases like limb-girdle muscular dystrophy and spinal muscular atrophy.

  5. Role of dystrophin in airway smooth muscle phenotype, contraction and lung function.

    Directory of Open Access Journals (Sweden)

    Pawan Sharma

    Full Text Available Dystrophin links the transmembrane dystrophin-glycoprotein complex to the actin cytoskeleton. We have shown that dystrophin-glycoprotein complex subunits are markers for airway smooth muscle phenotype maturation and together with caveolin-1, play an important role in calcium homeostasis. We tested if dystrophin affects phenotype maturation, tracheal contraction and lung physiology. We used dystrophin deficient Golden Retriever dogs (GRMD and mdx mice vs healthy control animals in our approach. We found significant reduction of contractile protein markers: smooth muscle myosin heavy chain (smMHC and calponin and reduced Ca2+ response to contractile agonist in dystrophin deficient cells. Immunocytochemistry revealed reduced stress fibers and number of smMHC positive cells in dystrophin-deficient cells, when compared to control. Immunoblot analysis of Akt1, GSK3β and mTOR phosphorylation further revealed that downstream PI3K signaling, which is essential for phenotype maturation, was suppressed in dystrophin deficient cell cultures. Tracheal rings from mdx mice showed significant reduction in the isometric contraction to methacholine (MCh when compared to genetic control BL10ScSnJ mice (wild-type. In vivo lung function studies using a small animal ventilator revealed a significant reduction in peak airway resistance induced by maximum concentrations of inhaled MCh in mdx mice, while there was no change in other lung function parameters. These data show that the lack of dystrophin is associated with a concomitant suppression of ASM cell phenotype maturation in vitro, ASM contraction ex vivo and lung function in vivo, indicating that a linkage between the DGC and the actin cytoskeleton via dystrophin is a determinant of the phenotype and functional properties of ASM.

  6. Dystrophin quantification and clinical correlations in Becker muscular dystrophy: implications for clinical trials.

    Science.gov (United States)

    Anthony, Karen; Cirak, Sebahattin; Torelli, Silvia; Tasca, Giorgio; Feng, Lucy; Arechavala-Gomeza, Virginia; Armaroli, Annarita; Guglieri, Michela; Straathof, Chiara S; Verschuuren, Jan J; Aartsma-Rus, Annemieke; Helderman-van den Enden, Paula; Bushby, Katherine; Straub, Volker; Sewry, Caroline; Ferlini, Alessandra; Ricci, Enzo; Morgan, Jennifer E; Muntoni, Francesco

    2011-12-01

    Duchenne muscular dystrophy is caused by mutations in the DMD gene that disrupt the open reading frame and prevent the full translation of its protein product, dystrophin. Restoration of the open reading frame and dystrophin production can be achieved by exon skipping using antisense oligonucleotides targeted to splicing elements. This approach aims to transform the Duchenne muscular dystrophy phenotype to that of the milder disorder, Becker muscular dystrophy, typically caused by in-frame dystrophin deletions that allow the production of an internally deleted but partially functional dystrophin. There is ongoing debate regarding the functional properties of the different internally deleted dystrophins produced by exon skipping for different mutations; more insight would be valuable to improve and better predict the outcome of exon skipping clinical trials. To this end, we have characterized the clinical phenotype of 17 patients with Becker muscular dystrophy harbouring in-frame deletions relevant to on-going or planned exon skipping clinical trials for Duchenne muscular dystrophy and correlated it to the levels of dystrophin, and dystrophin-associated protein expression. The cohort of 17 patients, selected exclusively on the basis of their genotype, included 4 asymptomatic, 12 mild and 1 severe patient. All patients had dystrophin levels of >40% of control and significantly higher dystrophin (P = 0.013), β-dystroglycan (P = 0.025) and neuronal nitric oxide synthase (P = 0.034) expression was observed in asymptomatic individuals versus symptomatic patients with Becker muscular dystrophy. Furthermore, grouping the patients by deletion, patients with Becker muscular dystrophy with deletions with an end-point of exon 51 (the skipping of which could rescue the largest group of Duchenne muscular dystrophy deletions) showed significantly higher dystrophin levels (P = 0.034) than those with deletions ending with exon 53. This is the first quantitative study on both

  7. Exon skipping and translation in patients with frameshift deletions in the dystrophin gene

    Energy Technology Data Exchange (ETDEWEB)

    Sherratt, T.G.; Dubowitz, V.; Sewry, C.A.; Strong, P.N. (Royal Postgraduate Medical School, London (United Kingdom)); Vulliamy, T. (Hammersmith Hospital, London (United Kingdom))

    1993-11-01

    Although many Duchenne muscular dystrophy patients have a deletion in the dystrophin gene which disrupts the translational reading frame, they express dystrophin in a small proportion of skeletal muscle fibers ([open quotes]revertant fibers[close quotes]). Antibody studies have shown, indirectly, that dystrophin synthesis in revertant fibers is facilitated by a frame-restoring mechanism; in the present study, the feasibility of mRNA splicing was investigated. Dystrophin transcripts were analyzed in skeletal muscle from individuals possessing revertant fibers and a frameshift deletion in the dystrophin gene. In each case a minor in-frame transcript was detected, in which exons adjacent to those deleted from the genome had been skipped. There appeared to be some correlation between the levels of in-frame transcripts and the predicted translation products. Low levels of alternatively spliced transcripts were also present in normal muscle. The results provide further evidence of exon skipping in the dystrophin gene and indicate that this may be involved in the synthesis of dystrophin by revertant fibers. 44 refs., 12 figs.

  8. Multiple Species Comparison of Cardiac Troponin T and Dystrophin: Unravelling the DNA behind Dilated Cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Jennifer England

    2017-07-01

    Full Text Available Animals have frequently been used as models for human disorders and mutations. Following advances in genetic testing and treatment options, and the decreasing cost of these technologies in the clinic, mutations in both companion and commercial animals are now being investigated. A recent review highlighted the genes associated with both human and non-human dilated cardiomyopathy. Cardiac troponin T and dystrophin were observed to be associated with both human and turkey (troponin T and canine (dystrophin dilated cardiomyopathies. This review gives an overview of the work carried out in cardiac troponin T and dystrophin to date in both human and animal dilated cardiomyopathy.

  9. Multiple Species Comparison of Cardiac Troponin T and Dystrophin: Unravelling the DNA behind Dilated Cardiomyopathy.

    Science.gov (United States)

    England, Jennifer; Loughna, Siobhan; Rutland, Catrin Sian

    2017-07-07

    Animals have frequently been used as models for human disorders and mutations. Following advances in genetic testing and treatment options, and the decreasing cost of these technologies in the clinic, mutations in both companion and commercial animals are now being investigated. A recent review highlighted the genes associated with both human and non-human dilated cardiomyopathy. Cardiac troponin T and dystrophin were observed to be associated with both human and turkey (troponin T) and canine (dystrophin) dilated cardiomyopathies. This review gives an overview of the work carried out in cardiac troponin T and dystrophin to date in both human and animal dilated cardiomyopathy.

  10. Structure of a WW domain-containing fragment of dystrophin complexed with {beta}-dystroglycan.

    Energy Technology Data Exchange (ETDEWEB)

    Huang, X.; Poy, F.; Zhang, R.; Joachimiak, A.; Sudol, M.; Eck, M. J.; Biosciences Division; Dana Farber Cancer Inst.; Harvard Medical School; Mount Sinai School of Medicine

    2000-08-01

    Dystrophin and {beta}-dystroglycan are components of the dystrophin--glycoprotein complex (DGC), a multimolecular assembly that spans the cell membrane and links the actin cytoskeleton to the extracellular basal lamina. Defects in the dystrophin gene are the cause of Duchenne and Becker muscular dystrophies. The C-terminal region of dystrophin binds the cytoplasmic tail of {beta}-dystroglycan, in part through the interaction of its WW domain with a proline-rich motif in the tail of {beta}-dystroglycan. Here we report the crystal structure of this portion of dystrophin in complex with the proline-rich binding site in {beta}-dystroglycan. The structure shows that the dystrophin WW domain is embedded in an adjacent helical region that contains two EF-hand-like domains. The {beta}-dystroglycan peptide binds a composite surface formed by the WW domain and one of these EF-hands. Additionally, the structure reveals striking similarities in the mechanisms of proline recognition employed by WW domains and SH3 domains.

  11. Muscular dystrophy in a family of Labrador Retrievers with no muscle dystrophin and a mild phenotype.

    Science.gov (United States)

    Vieira, Natassia M; Guo, Ling T; Estrela, Elicia; Kunkel, Louis M; Zatz, Mayana; Shelton, G Diane

    2015-05-01

    Animal models of dystrophin deficient muscular dystrophy, most notably canine X-linked muscular dystrophy, play an important role in developing new therapies for human Duchenne muscular dystrophy. Although the canine disease is a model of the human disease, the variable severity of clinical presentations in the canine may be problematic for pre-clinical trials, but also informative. Here we describe a family of Labrador Retrievers with three generations of male dogs having markedly increased serum creatine kinase activity, absence of membrane dystrophin, but with undetectable clinical signs of muscle weakness. Clinically normal young male Labrador Retriever puppies were evaluated prior to surgical neuter by screening laboratory blood work, including serum creatine kinase activity. Serum creatine kinase activities were markedly increased in the absence of clinical signs of muscle weakness. Evaluation of muscle biopsies confirmed a dystrophic phenotype with both degeneration and regeneration. Further evaluations by immunofluorescence and western blot analysis confirmed the absence of muscle dystrophin. Although dystrophin was not identified in the muscles, we did not find any detectable deletions or duplications in the dystrophin gene. Sequencing is now ongoing to search for point mutations. Our findings in this family of Labrador Retriever dogs lend support to the hypothesis that, in exceptional situations, muscle with no dystrophin may be functional. Unlocking the secrets that protect these dogs from a severe clinical myopathy is a great challenge which may have important implications for future treatment of human muscular dystrophies. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Computational study of the human dystrophin repeats: interaction properties and molecular dynamics.

    Directory of Open Access Journals (Sweden)

    Baptiste Legrand

    Full Text Available Dystrophin is a large protein involved in the rare genetic disease Duchenne muscular dystrophy (DMD. It functions as a mechanical linker between the cytoskeleton and the sarcolemma, and is able to resist shear stresses during muscle activity. In all, 75% of the dystrophin molecule consists of a large central rod domain made up of 24 repeat units that share high structural homology with spectrin-like repeats. However, in the absence of any high-resolution structure of these repeats, the molecular basis of dystrophin central domain's functions has not yet been deciphered. In this context, we have performed a computational study of the whole dystrophin central rod domain based on the rational homology modeling of successive and overlapping tandem repeats and the analysis of their surface properties. Each tandem repeat has very specific surface properties that make it unique. However, the repeats share enough electrostatic-surface similarities to be grouped into four separate clusters. Molecular dynamics simulations of four representative tandem repeats reveal specific flexibility or bending properties depending on the repeat sequence. We thus suggest that the dystrophin central rod domain is constituted of seven biologically relevant sub-domains. Our results provide evidence for the role of the dystrophin central rod domain as a scaffold platform with a wide range of surface features and biophysical properties allowing it to interact with its various known partners such as proteins and membrane lipids. This new integrative view is strongly supported by the previous experimental works that investigated the isolated domains and the observed heterogeneity of the severity of dystrophin related pathologies, especially Becker muscular dystrophy.

  13. Early dystrophin loss is coincident with the transition of compensated cardiac hypertrophy to heart failure.

    Directory of Open Access Journals (Sweden)

    Fernanda P Prado

    Full Text Available Hypertension causes cardiac hypertrophy, one of the most important risk factors for heart failure (HF. Despite the importance of cardiac hypertrophy as a risk factor for the development of HF, not all hypertrophied hearts will ultimately fail. Alterations of cytoskeletal and sarcolemma-associated proteins are considered markers cardiac remodeling during HF. Dystrophin provides mechanical stability to the plasma membrane through its interactions with the actin cytoskeleton and, indirectly, to extracellular matrix proteins. This study was undertaken to evaluate dystrophin and calpain-1 in the transition from compensated cardiac hypertrophy to HF. Wistar rats were subjected to abdominal aorta constriction and killed at 30, 60 and 90 days post surgery (dps. Cardiac function and blood pressure were evaluated. The hearts were collected and Western blotting and immunofluorescence performed for dystrophin, calpain-1, alpha-fodrin and calpastatin. Statistical analyses were performed and considered significant when p<0.05. After 90 dps, 70% of the animals showed hypertrophic hearts (HH and 30% hypertrophic+dilated hearts (HD. Systolic and diastolic functions were preserved at 30 and 60 dps, however, decreased in the HD group. Blood pressure, cardiomyocyte diameter and collagen content were increased at all time points. Dystrophin expression was lightly increased at 30 and 60 dps and HH group. HD group showed decreased expression of dystrophin and calpastatin and increased expression of calpain-1 and alpha-fodrin fragments. The first signals of dystrophin reduction were observed as early as 60 dps. In conclusion, some hearts present a distinct molecular pattern at an early stage of the disease; this pattern could provide an opportunity to identify these failure-prone hearts during the development of the cardiac disease. We showed that decreased expression of dystrophin and increased expression of calpains are coincident and could work as possible

  14. Novel dystrophin mutations revealed by analysis of dystrophin mRNA: alternative splicing suppresses the phenotypic effect of a nonsense mutation

    Czech Academy of Sciences Publication Activity Database

    Fajkusová, L.; Lukáš, Z.; Tvrdíková, M.; Kuhrová, V.; Hájek, J.; Fajkus, Jiří

    2001-01-01

    Roč. 11, č. 2 (2001), s. 133-138 ISSN 0960-8966 R&D Projects: GA MZd IZ3700; GA MZd NM19; GA MZd NA5227 Institutional research plan: CEZ:AV0Z5004920 Keywords : Duchenne muscular dystrophy * Becker muscular dystrophy * dystrophin mRNA Subject RIV: BO - Biophysics Impact factor: 2.547, year: 2001

  15. Mismatched single stranded antisense oligonucleotides can induce efficient dystrophin splice switching

    Directory of Open Access Journals (Sweden)

    Kole Ryszard

    2011-10-01

    Full Text Available Abstract Background Antisense oligomer induced exon skipping aims to reduce the severity of Duchenne muscular dystrophy by redirecting splicing during pre-RNA processing such that the causative mutation is by-passed and a shorter but partially functional Becker muscular dystrophy-like dystrophin isoform is produced. Normal exons are generally targeted to restore the dystrophin reading frame however, an appreciable subset of dystrophin mutations are intra-exonic and therefore have the potential to compromise oligomer efficiency, necessitating personalised oligomer design for some patients. Although antisense oligomers are easily personalised, it remains unclear whether all patient polymorphisms within antisense oligomer target sequences will require the costly process of producing and validating patient specific compounds. Methods Here we report preclinical testing of a panel of splice switching antisense oligomers, designed to excise exon 25 from the dystrophin transcript, in normal and dystrophic patient cells. These patient cells harbour a single base insertion in exon 25 that lies within the target sequence of an oligomer shown to be effective at removing exon 25. Results It was anticipated that such a mutation would compromise oligomer binding and efficiency. However, we show that, despite the mismatch an oligomer, designed and optimised to excise exon 25 from the normal dystrophin mRNA, removes the mutated exon 25 more efficiently than the mutation-specific oligomer. Conclusion This raises the possibility that mismatched AOs could still be therapeutically applicable in some cases, negating the necessity to produce patient-specific compounds.

  16. Functional rescue of dystrophin-deficient mdx mice by a chimeric peptide-PMO.

    Science.gov (United States)

    Yin, Haifang; Moulton, Hong M; Betts, Corinne; Merritt, Thomas; Seow, Yiqi; Ashraf, Shirin; Wang, Qingsong; Boutilier, Jordan; Wood, Matthew Ja

    2010-10-01

    Splice modulation using antisense oligonucleotides (AOs) has been shown to yield targeted exon exclusion to restore the open reading frame and generate truncated but partially functional dystrophin protein. This has been successfully demonstrated in dystrophin-deficient mdx mice and in Duchenne muscular dystrophy (DMD) patients. However, DMD is a systemic disease; successful therapeutic exploitation of this approach will therefore depend on effective systemic delivery of AOs to all affected tissues. We have previously shown the potential of a muscle-specific/arginine-rich chimeric peptide-phosphorodiamidate morpholino (PMO) conjugate, but its long-term activity, optimized dosing regimen, capacity for functional correction and safety profile remain to be established. Here, we report the results of this chimeric peptide-PMO conjugate in the mdx mouse using low doses (3 and 6 mg/kg) administered via a 6 biweekly systemic intravenous injection protocol. We show 100% dystrophin-positive fibers and near complete correction of the dystrophin transcript defect in all peripheral muscle groups, with restoration of 50% dystrophin protein over 12 weeks, leading to correction of the DMD pathological phenotype and restoration of muscle function in the absence of detectable toxicity or immune response. Chimeric muscle-specific/cell-penetrating peptides therefore represent highly promising agents for systemic delivery of splice-correcting PMO oligomers for DMD therapy.

  17. Deficiency in Cardiac Dystrophin Affects the Abundance of the α-/β-Dystroglycan Complex

    Directory of Open Access Journals (Sweden)

    James Lohan

    2005-01-01

    Full Text Available Although Duchenne muscular dystrophy is primarily categorised as a skeletal muscle disease, deficiency in the membrane cytoskeletal protein dystrophin also affects the heart. The central transsarcolemmal linker between the actin membrane cytoskeleton and the extracellular matrix is represented by the dystrophin-associated dystroglycans. Chemical cross-linking analysis revealed no significant differences in the dimeric status of the α-/β-dystroglycan subcomplex in the dystrophic mdx heart as compared to normal cardiac tissue. In analogy to skeletal muscle fibres, heart muscle also exhibited a greatly reduced abundance of both dystroglycans in dystrophin-deficient cells. Immunoblotting demonstrated that the degree of reduction in α-dystroglycan is more pronounced in matured mdx skeletal muscle as contrasted to the mdx heart. The fact that the deficiency in dystrophin triggers a similar pathobiochemical response in both types of muscle suggests that the cardiomyopathic complications observed in x-linked muscular dystrophy might be initiated by the loss of the dystrophin-associated surface glycoprotein complex.

  18. Decreased inward rectifier potassium current IK1 in dystrophin-deficient ventricular cardiomyocytes.

    Science.gov (United States)

    Rubi, Lena; Koenig, Xaver; Kubista, Helmut; Todt, Hannes; Hilber, Karlheinz

    2017-03-04

    Kir2.x channels in ventricular cardiomyocytes (most prominently Kir2.1) account for the inward rectifier potassium current I K1 , which controls the resting membrane potential and the final phase of action potential repolarization. Recently it was hypothesized that the dystrophin-associated protein complex (DAPC) is important in the regulation of Kir2.x channels. To test this hypothesis, we investigated potential I K1 abnormalities in dystrophin-deficient ventricular cardiomyocytes derived from the hearts of Duchenne muscular dystrophy mouse models. We found that I K1 was substantially diminished in dystrophin-deficient cardiomyocytes when compared to wild type myocytes. This finding represents the first functional evidence for a significant role of the DAPC in the regulation of Kir2.x channels.

  19. Heteroduplex analysis of the dystrophin gene: Application to point mutation and carrier detection

    Energy Technology Data Exchange (ETDEWEB)

    Prior, T.W.; Papp, A.C.; Snyder, P.J.; Sedra, M.S.; Western, L.M.; Bartolo, C.; Mendell, J.R. [Ohio State Univ., Columbus, OH (United States); Moxley, R.T. [Univ. of Rochester Medical Center, NY (United States)

    1994-03-01

    Approximately one-third of Duchenne muscular dystrophy patients have undefined mutations in the dystrophin gene. For carrier and prenatal studies in families without detectable mutations, the indirect restriction fragment length polymorphism linkage approach is used. Using a multiplex amplification and heteroduplex analysis of dystrophin exons, the authors identified nonsense mutations in two DMD patients. Although the nonsense mutations are predicted to severely truncate the dystrophin protein, both patients presented with mild clinical courses of the disease. As a result of identifying the mutation in the affected boys, direct carrier studies by heteroduplex analysis were extended to other relatives. The authors conclude that the technique is not only ideal for mutation detection but is also useful for diagnostic testing. 29 refs., 4 figs.

  20. Detection of new paternal dystrophin gene mutations in isolated cases of dystrophinopathy in females

    Energy Technology Data Exchange (ETDEWEB)

    Pegoraro, E.; Wessel, H.B.; Schwartz, L.; Hoffman, E.P. (Univ. of Pittsburgh, PA (United States)); Schimke, R.N. (Kansas Univ. Medical Center, Kansas City (United States)); Arahata, Kiichi; Hayashi, Yukiko (National Institute of Neurosciences, Tokyo (Japan)); Stern, H. (Children' s National Medical Center, Washington, DC (United States)); Marks, H. (A.I. duPont Institute, Wilmington (United States)); Glasberg, M.R. (Henry Ford Hospital, Detroit, MI (United States)) (and others)

    1994-06-01

    Duchenne muscular dystrophy is one of the most common lethal monogenic disorders and is caused by dystrophin deficiency. The disease is transmitted as an X-linked recessive trait; however, recent biochemical and clinical studies have shown that many girls and women with a primary myopathy have an underlying dystrophinopathy, despite a negative family history for Duchenne dystrophy. These isolated female dystrophinopathy patients carried ambiguous diagnoses with presumed autosomal recessive inheritance (limb-girdle muscular dystrophy) prior to biochemical detection of dystrophin abnormalities in their muscle biopsy. It has been assumed that these female dystrophinopathy patients are heterozygous carries who show preferential inactivation of the X chromosome harboring the normal dystrophin gene, although this has been shown for only a few X:autosome translocations and for two cases of discordant monozygotic twin female carriers. Here the authors study X-inactivation patterns of 13 female dystrophinopathy patients - 10 isolated cases and 3 cases with a positive family history for Duchenne dystrophy in males. They show that all cases have skewed X-inactivation patterns in peripheral blood DNA. Of the nine isolated cases informative in the assay, eight showed inheritance of the dystrophin gene mutation from the paternal germ line. Only a single case showed maternal inheritance. The 10-fold higher incidence of paternal transmission of dystrophin gene mutations in these cases is at 30-fold variance with Bayesian predictions and gene mutation rates. Thus, the results suggest some mechanistic interaction between new dystrophin gene mutations, paternal inheritance, and skewed X inactivation. The results provide both empirical risk data and a molecular diagnostic test method, which permit genetic counseling and prenatal diagnosis of this new category of patients. 58 refs., 7 figs., 2 tabs.

  1. Dystrophin Expressing Chimeric (DEC) Human Cells Provide a Potential Therapy for Duchenne Muscular Dystrophy.

    Science.gov (United States)

    Siemionow, Maria; Cwykiel, Joanna; Heydemann, Ahlke; Garcia, Jesus; Marchese, Enza; Siemionow, Krzysztof; Szilagyi, Erzsebet

    2018-06-01

    Duchenne Muscular Dystrophy (DMD) is a progressive and lethal disease caused by mutations of the dystrophin gene. Currently no cure exists. Stem cell therapies targeting DMD are challenged by limited engraftment and rejection despite the use of immunosuppression. There is an urgent need to introduce new stem cell-based therapies that exhibit low allogenic profiles and improved cell engraftment. In this proof-of-concept study, we develop and test a new human stem cell-based approach to increase engraftment, limit rejection, and restore dystrophin expression in the mdx/scid mouse model of DMD. We introduce two Dystrophin Expressing Chimeric (DEC) cell lines created by ex vivo fusion of human myoblasts (MB) derived from two normal donors (MB N1 /MB N2 ), and normal and DMD donors (MB N /MB DMD ). The efficacy of fusion was confirmed by flow cytometry and confocal microscopy based on donor cell fluorescent labeling (PKH26/PKH67). In vitro, DEC displayed phenotype and genotype of donor parent cells, expressed dystrophin, and maintained proliferation and myogenic differentiation. In vivo, local delivery of both DEC lines (0.5 × 10 6 ) restored dystrophin expression (17.27%±8.05-MB N1 /MB N2 and 23.79%±3.82-MB N /MB DMD ) which correlated with significant improvement of muscle force, contraction and tolerance to fatigue at 90 days after DEC transplant to the gastrocnemius muscles (GM) of dystrophin-deficient mdx/scid mice. This study establishes DEC as a potential therapy for DMD and other types of muscular dystrophies.

  2. Use of epitope libraries to identify exon-specific monoclonal antibodies for characterization of altered dystrophins in muscular dystrophy

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen thi Man; Morris, G.E. (North East Wales Inst., Clwyd (United Kingdom))

    1993-06-01

    The majority of mutations in Xp21-linked muscular dystrophy (MD) can be identified by PCR or Southern blotting, as deletions or duplications of groups of exons in the dystrophin gene, but it is not always possible to predict how much altered dystrophin, if any, will be produced. Use of exon-specific monoclonal antibodies (mAbs) on muscle biopsies from MD patients can, in principle, provide information on both the amount of altered dystrophin produced and, when dystrophin is present, the nature of the genetic deletion or point mutation. For this purpose, mAbs which recognize regions of dystrophin encoded by known exons and whose binding is unaffected by the absence of adjacent exons are required. To map mAbs to specific exons, random [open quotes]libraries[close quotes] of expressed dystrophin fragments were created by cloning DNAseI digestion fragments of a 4.3-kb dystrophin cDNA into a pTEX expression vector. The libraries were then used to locate the epitopes recognized by 48 mAbs to fragments of 25--60 amino acids within the 1,434-amino-acid dystrophin fragment used to produce the antibodies. This is sufficiently detailed to allow further refinement by using synthetic peptides and, in many cases, to identify the exon in the DMD (Duchenne MD) gene which encodes the epitope. To illustrate their use in dystrophin analysis, a Duchenne patient with a frameshift deletion of exons 42 and 43 makes a truncated dystrophin encoded by exons 1--41, and the authors now show that this can be detected in the sarcolemma by mAbs up to and including those specific for exon 41 epitopes but not by mAbs specific for exon 43 or later epitopes. 38 refs., 2 figs., 4 tabs.

  3. Dystrophin deficiency leads to disturbance of LAMP1-vesicle-associated protein secretion

    DEFF Research Database (Denmark)

    Duguez, S.; Duddy, W.; Johnston, H.

    2013-01-01

    Duchenne muscular dystrophy results from loss of the protein dystrophin, which links the intracellular cytoskeletal network with the extracellular matrix, but deficiency in this function does not fully explain the onset or progression of the disease. While some intracellular events involved...... in the degeneration of dystrophin-deficient muscle fibers have been well characterized, changes in their secretory profile are undescribed. To analyze the secretome profile of mdx myotubes independently of myonecrosis, we labeled the proteins of mdx and wild-type myotubes with stable isotope-labeled amino acids...

  4. The polyproline site in hinge 2 influences the functional capacity of truncated dystrophins.

    Directory of Open Access Journals (Sweden)

    Glen B Banks

    2010-05-01

    Full Text Available Mutations in dystrophin can lead to Duchenne muscular dystrophy or the more mild form of the disease, Becker muscular dystrophy. The hinge 3 region in the rod domain of dystrophin is particularly prone to deletion mutations. In-frame deletions of hinge 3 are predicted to lead to BMD, however the severity of disease can vary considerably. Here we performed extensive structure-function analyses of truncated dystrophins with modified hinges and spectrin-like repeats in mdx mice. We found that the polyproline site in hinge 2 profoundly influences the functional capacity of a microdystrophin(DeltaR4-R23/DeltaCT with a large deletion in the hinge 3 region. Inclusion of polyproline in microdystrophin(DeltaR4-R23/DeltaCT led to small myofibers (12% smaller than wild-type, Achilles myotendinous disruption, ringed fibers, and aberrant neuromuscular junctions in the mdx gastrocnemius muscles. Replacing hinge 2 of microdystrophin(DeltaR4-R23/DeltaCT with hinge 3 significantly improved the functional capacity to prevent muscle degeneration, increase muscle fiber area, and maintain the junctions. We conclude that the rigid alpha-helical structure of the polyproline site significantly impairs the functional capacity of truncated dystrophins to maintain appropriate connections between the cytoskeleton and extracellular matrix.

  5. Studying the role of dystrophin-associated proteins in influencing Becker muscular dystrophy disease severity.

    Science.gov (United States)

    van den Bergen, J C; Wokke, B H A; Hulsker, M A; Verschuuren, J J G M; Aartsma-Rus, A M

    2015-03-01

    Becker muscular dystrophy is characterized by a variable disease course. Many factors have been implicated to contribute to this diversity, among which the expression of several components of the dystrophin associated glycoprotein complex. Together with dystrophin, most of these proteins anchor the muscle fiber cytoskeleton to the extracellular matrix, thus protecting the muscle from contraction induced injury, while nNOS is primarily involved in inducing vasodilation during muscle contraction, enabling adequate muscle oxygenation. In the current study, we investigated the role of three components of the dystrophin associated glycoprotein complex (beta-dystroglycan, gamma-sarcoglycan and nNOS) and the dystrophin homologue utrophin on disease severity in Becker patients. Strength measurements, data about disease course and fresh muscle biopsies of the anterior tibial muscle were obtained from 24 Becker patients aged 19 to 66. The designation of Becker muscular dystrophy in this study was based on the mutation and not on the clinical severity. Contrary to previous studies, we were unable to find a relationship between expression of nNOS, beta-dystroglycan and gamma-sarcoglycan at the sarcolemma and disease severity, as measured by muscle strength in five muscle groups and age at reaching several disease milestones. Unexpectedly, we found an inverse correlation between utrophin expression at the sarcolemma and age at reaching disease milestones. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Analysis of Dystrophin Gene Deletions by Multiplex PCR in Moroccan Patients

    Directory of Open Access Journals (Sweden)

    Aziza Sbiti

    2002-01-01

    Full Text Available Duchenne and Becker muscular dystrophy (DMD and BMD are X-linked diseases resulting from a defect in the dystrophin gene located on Xp21. DMD is the most frequent neuromuscular disease in humans (1/3500 male newborn. Deletions in the dystrophin gene represent 65% of mutations in DMD/BMD patients. We have analyzed DNA from 72 Moroccan patients with DMD/BMD using the multiplex polymerase chain reaction (PCR to screen for exon deletions within the dystrophin gene, and to estimate the frequency of these abnormalities. We found dystrophin gene deletions in 37 cases. Therefore the frequency in Moroccan DMD/BMD patients is about 51.3%. All deletions were clustered in the two known hot-spots regions, and in 81% of cases deletions were detected in the region from exon 43 to exon 52. These findings are comparable to those reported in other studies. It is important to note that in our population, we can first search for deletions of DMD gene in the most frequently deleted exons determined by this study. This may facilitate the molecular diagnosis of DMD and BMD in our country.

  7. Targeted Exon Skipping to Correct Exon Duplications in the Dystrophin Gene

    Directory of Open Access Journals (Sweden)

    Kane L Greer

    2014-01-01

    Full Text Available Duchenne muscular dystrophy is a severe muscle-wasting disease caused by mutations in the dystrophin gene that ablate functional protein expression. Although exonic deletions are the most common Duchenne muscular dystrophy lesion, duplications account for 10–15% of reported disease-causing mutations, and exon 2 is the most commonly duplicated exon. Here, we describe the in vitro evaluation of phosphorodiamidate morpholino oligomers coupled to a cell-penetrating peptide and 2′-O-methyl phosphorothioate oligonucleotides, using three distinct strategies to reframe the dystrophin transcript in patient cells carrying an exon 2 duplication. Differences in exon-skipping efficiencies in vitro were observed between oligomer analogues of the same sequence, with the phosphorodiamidate morpholino oligomer coupled to a cell-penetrating peptide proving the most effective. Differences in exon 2 excision efficiency between normal and exon 2 duplication cells, were apparent, indicating that exon context influences oligomer-induced splice switching. Skipping of a single copy of exon 2 was induced in the cells carrying an exon 2 duplication, the simplest strategy to restore the reading frame and generate a normal dystrophin transcript. In contrast, multiexon skipping of exons 2–7 to generate a Becker muscular dystrophy-like dystrophin transcript was more challenging and could only be induced efficiently with the phosphorodiamidate morpholino oligomer chemistry.

  8. Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes

    Directory of Open Access Journals (Sweden)

    Susi Zatti

    2014-01-01

    Full Text Available Duchenne muscular dystrophy (DMD–associated cardiac diseases are emerging as a major cause of morbidity and mortality in DMD patients, and many therapies for treatment of skeletal muscle failed to improve cardiac function. The reprogramming of patients' somatic cells into pluripotent stem cells, combined with technologies for correcting the genetic defect, possesses great potential for the development of new treatments for genetic diseases. In this study, we obtained human cardiomyocytes from DMD patient–derived, induced pluripotent stem cells genetically corrected with a human artificial chromosome carrying the whole dystrophin genomic sequence. Stimulation by cytokines was combined with cell culturing on hydrogel with physiological stiffness, allowing an adhesion-dependent maturation and a proper dystrophin expression. The obtained cardiomyocytes showed remarkable sarcomeric organization of cardiac troponin T and α-actinin, expressed cardiac-specific markers, and displayed electrically induced calcium transients lasting less than 1 second. We demonstrated that the human artificial chromosome carrying the whole dystrophin genomic sequence is stably maintained throughout the cardiac differentiation process and that multiple promoters of the dystrophin gene are properly activated, driving expression of different isoforms. These dystrophic cardiomyocytes can be a valuable source for in vitro modeling of DMD-associated cardiac disease. Furthermore, the derivation of genetically corrected, patient-specific cardiomyocytes represents a step toward the development of innovative cell and gene therapy approaches for DMD.

  9. Comparative Analysis of Vertebrate Dystrophin Loci Indicate Intron Gigantism as a Common Feature

    Science.gov (United States)

    Pozzoli, Uberto; Elgar, Greg; Cagliani, Rachele; Riva, Laura; Comi, Giacomo P.; Bresolin, Nereo; Bardoni, Alessandra; Sironi, Manuela

    2003-01-01

    The human DMD gene is the largest known to date, spanning > 2000 kb on the X chromosome. The gene size is mainly accounted for by huge intronic regions. We sequenced 190 kb of Fugu rubripes (pufferfish) genomic DNA corresponding to the complete dystrophin gene (FrDMD) and provide the first report of gene structure and sequence comparison among dystrophin genomic sequences from different vertebrate organisms. Almost all intron positions and phases are conserved between FrDMD and its mammalian counterparts, and the predicted protein product of the Fugu gene displays 55% identity and 71% similarity to human dystrophin. In analogy to the human gene, FrDMD presents several-fold longer than average intronic regions. Analysis of intron sequences of the human and murine genes revealed that they are extremely conserved in size and that a similar fraction of total intron length is represented by repetitive elements; moreover, our data indicate that intron expansion through repeat accumulation in the two orthologs is the result of independent insertional events. The hypothesis that intron length might be functionally relevant to the DMD gene regulation is proposed and substantiated by the finding that dystrophin intron gigantism is common to the three vertebrate genes. [Supplemental material is available online at www.genome.org.] PMID:12727896

  10. Skeletal Muscle Differentiation on a Chip Shows Human Donor Mesoangioblasts' Efficiency in Restoring Dystrophin in a Duchenne Muscular Dystrophy Model.

    Science.gov (United States)

    Serena, Elena; Zatti, Susi; Zoso, Alice; Lo Verso, Francesca; Tedesco, F Saverio; Cossu, Giulio; Elvassore, Nicola

    2016-12-01

    : Restoration of the protein dystrophin on muscle membrane is the goal of many research lines aimed at curing Duchenne muscular dystrophy (DMD). Results of ongoing preclinical and clinical trials suggest that partial restoration of dystrophin might be sufficient to significantly reduce muscle damage. Different myogenic progenitors are candidates for cell therapy of muscular dystrophies, but only satellite cells and pericytes have already entered clinical experimentation. This study aimed to provide in vitro quantitative evidence of the ability of mesoangioblasts to restore dystrophin, in terms of protein accumulation and distribution, within myotubes derived from DMD patients, using a microengineered model. We designed an ad hoc experimental strategy to miniaturize on a chip the standard process of muscle regeneration independent of variables such as inflammation and fibrosis. It is based on the coculture, at different ratios, of human dystrophin-positive myogenic progenitors and dystrophin-negative myoblasts in a substrate with muscle-like physiological stiffness and cell micropatterns. Results showed that both healthy myoblasts and mesoangioblasts restored dystrophin expression in DMD myotubes. However, mesoangioblasts showed unexpected efficiency with respect to myoblasts in dystrophin production in terms of the amount of protein produced (40% vs. 15%) and length of the dystrophin membrane domain (210-240 µm vs. 40-70 µm). These results show that our microscaled in vitro model of human DMD skeletal muscle validated previous in vivo preclinical work and may be used to predict efficacy of new methods aimed at enhancing dystrophin accumulation and distribution before they are tested in vivo, reducing time, costs, and variability of clinical experimentation. This study aimed to provide in vitro quantitative evidence of the ability of human mesoangioblasts to restore dystrophin, in terms of protein accumulation and distribution, within myotubes derived from

  11. Ex vivo gene editing of the dystrophin gene in muscle stem cells mediated by peptide nucleic acid single stranded oligodeoxynucleotides induces stable expression of dystrophin in a mouse model for Duchenne muscular dystrophy.

    Science.gov (United States)

    Nik-Ahd, Farnoosh; Bertoni, Carmen

    2014-07-01

    Duchenne muscular dystrophy (DMD) is a fatal disease caused by mutations in the dystrophin gene, which result in the complete absence of dystrophin protein throughout the body. Gene correction strategies hold promise to treating DMD. Our laboratory has previously demonstrated the ability of peptide nucleic acid single-stranded oligodeoxynucleotides (PNA-ssODNs) to permanently correct single-point mutations at the genomic level. In this study, we show that PNA-ssODNs can target and correct muscle satellite cells (SCs), a population of stem cells capable of self-renewing and differentiating into muscle fibers. When transplanted into skeletal muscles, SCs transfected with correcting PNA-ssODNs were able to engraft and to restore dystrophin expression. The number of dystrophin-positive fibers was shown to significantly increase over time. Expression was confirmed to be the result of the activation of a subpopulation of SCs that had undergone repair as demonstrated by immunofluorescence analyses of engrafted muscles using antibodies specific to full-length dystrophin transcripts and by genomic DNA analysis of dystrophin-positive fibers. Furthermore, the increase in dystrophin expression detected over time resulted in a significant improvement in muscle morphology. The ability of transplanted cells to return into quiescence and to activate upon demand was confirmed in all engrafted muscles following injury. These results demonstrate the feasibility of using gene editing strategies to target and correct SCs and further establish the therapeutic potential of this approach to permanently restore dystrophin expression into muscle of DMD patients. © 2014 AlphaMed Press.

  12. A Translational Pathway Toward a Clinical Trial Using the Second-Generation AAV Micro Dystrophin Vector

    Science.gov (United States)

    2017-09-01

    American Society of Gene & Cell Therapy. Washington, DC May 10-13, 2017 4) Chady H. Hakim, Kasun Kodippili, Gregory Jenkins, Hsiao T. Yang...translational studies for developing an obese and diabetic ossabaw swine model of HFpEF. Regulation of Work Capacity in Cardiac Myocytes McDonald, PI 5...Fellowship (Olver, PI) 0% effort, Emter, Supervising PI American Heart Association 1/1/2016-12/31/2017 $50,000 total funding Major goals: Salary

  13. Myoblots: dystrophin quantification by in-cell western assay for a streamlined development of Duchenne muscular dystrophy (DMD) treatments.

    Science.gov (United States)

    Ruiz-Del-Yerro, E; Garcia-Jimenez, I; Mamchaoui, K; Arechavala-Gomeza, V

    2017-10-31

    New therapies for neuromuscular disorders are often mutation specific and require to be studied in patient's cell cultures. In Duchenne muscular dystrophy (DMD) dystrophin restoration drugs are being developed but as muscle cell cultures from DMD patients are scarce and do not grow or differentiate well, only a limited number of candidate drugs are tested. Moreover, dystrophin quantification by western blotting requires a large number of cultured cells; so fewer compounds are as thoroughly screened as is desirable. We aimed to develop a quantitative assessment tool using fewer cells to contribute in the study of dystrophin and to identify better drug candidates. An 'in-cell western' assay is a quantitative immunofluorescence assay performed in cell culture microplates that allows protein quantification directly in culture, allowing a higher number of experimental repeats and throughput. We have optimized the assay ('myoblot') to be applied to the study of differentiated myoblast cultures. After an exhaustive optimization of the technique to adapt it to the growth and differentiation rates of our cultures and the low intrinsic expression of our proteins of interests, our myoblot protocol allows the quantification of dystrophin and other muscle-associated proteins in muscle cell cultures. We are able to distinguish accurately between the different sets of patients based on their dystrophin expression and detect dystrophin restoration after treatment. We expect that this new tool to quantify muscle proteins in DMD and other muscle disorders will aid in their diagnosis and in the development of new therapies. © 2017 British Neuropathological Society.

  14. Assessment of the structural and functional impact of in-frame mutations of the DMD gene, using the tools included in the eDystrophin online database

    Directory of Open Access Journals (Sweden)

    Nicolas Aurélie

    2012-07-01

    Full Text Available Abstract Background Dystrophin is a large essential protein of skeletal and heart muscle. It is a filamentous scaffolding protein with numerous binding domains. Mutations in the DMD gene, which encodes dystrophin, mostly result in the deletion of one or several exons and cause Duchenne (DMD and Becker (BMD muscular dystrophies. The most common DMD mutations are frameshift mutations resulting in an absence of dystrophin from tissues. In-frame DMD mutations are less frequent and result in a protein with partial wild-type dystrophin function. The aim of this study was to highlight structural and functional modifications of dystrophin caused by in-frame mutations. Methods and results We developed a dedicated database for dystrophin, the eDystrophin database. It contains 209 different non frame-shifting mutations found in 945 patients from a French cohort and previous studies. Bioinformatics tools provide models of the three-dimensional structure of the protein at deletion sites, making it possible to determine whether the mutated protein retains the typical filamentous structure of dystrophin. An analysis of the structure of mutated dystrophin molecules showed that hybrid repeats were reconstituted at the deletion site in some cases. These hybrid repeats harbored the typical triple coiled-coil structure of native repeats, which may be correlated with better function in muscle cells. Conclusion This new database focuses on the dystrophin protein and its modification due to in-frame deletions in BMD patients. The observation of hybrid repeat reconstitution in some cases provides insight into phenotype-genotype correlations in dystrophin diseases and possible strategies for gene therapy. The eDystrophin database is freely available: http://edystrophin.genouest.org/.

  15. Duchenne muscular dystrophy diagnosed by dystrophin gene deletion test: A case report

    Directory of Open Access Journals (Sweden)

    Rathod Kishor G, Dawre Rahul M, Kamble Milind B,Tambe Saleem H

    2014-04-01

    Full Text Available Duchenne muscular dystrophy (DMD is an X-linked recessive disease affecting 1 in 3600—6000 live male births. A muscle biopsy is not necessary if a genetic diagnosis is secured first, particularly as some families might view the procedure as traumatic. DMD occurs as a result of mutations (mainly deletions in the dystrophin gene (DMD; locus Xp21.2. Mutations lead to an absence of or defect in the protein dystrophin, which results in progressive muscle degeneration leading to loss of independent ambulation. Ninety percent of out frame mutations result in DMD, while 90% of in-frame mutations result in BMD. Electron microscopy is not required to confirm DMD. Genetic testing is mandatory irrespective of biopsy results. But the muscle biopsy is not required if the diagnosis is secured first by genetic testing.

  16. Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle

    DEFF Research Database (Denmark)

    Hjortkjær, Camilla Brolin; Shiraishi, Takehiko; Hojman, Pernille

    2015-01-01

    for improvement of in vivo cellular availability, we have investigated the effect of electrotransfer upon intramuscular (i.m.) PNA administration in vivo. Antisense PNA targeting exon 23 of the murine dystrophin gene was administered by i.m. injection to the tibialis anterior (TA) muscle of normal NMRI......Peptide nucleic acid (PNA) is a synthetic DNA mimic that has shown potential for discovery of novel splice switching antisense drugs. However, in vivo cellular delivery has been a limiting factor for development, and only few successful studies have been reported. As a possible modality...... switching was detected at the RNA level up to 4 weeks after a single-dose treatment. In dystrophic muscles of the MDX mouse, electroporation increased the number of dystrophin-positive fibers about 2.5-fold at 2 weeks after a single PNA administration compared to injection only. In conclusion, we find...

  17. Dystroglycan and muscular dystrophies related to the dystrophin-glycoprotein complex.

    Science.gov (United States)

    Sciandra, Francesca; Bozzi, Manuela; Bianchi, Marzia; Pavoni, Ernesto; Giardina, Bruno; Brancaccio, Andrea

    2003-01-01

    Dystroglycan (DG) is an adhesion molecule composed of two subunits, alpha and beta, that are produced by the post-translational cleavage of a single precursor molecule. DG is a pivotal component of the dystrophin-glycoprotein complex (DGC), which connects the extracellular matrix to the cytoskeleton in skeletal muscle and many other tissues. Some muscular dystrophies are caused by mutations of DGC components, such as dystrophin, sarcoglycan or laminin-2, or also of DGC-associated molecules, such as caveolin-3. DG-null mice died during early embriogenesis and no neuromuscular diseases directly associated to genetic abnormalities of DG were identified so far. However, DG plays a crucial role for muscle integrity since its targeting at the sarcolemma is often perturbed in DGC-related neuromuscular disorders.

  18. Restoration of half the normal dystrophin sequence in a double-deletion Duchenne muscular dystrophy family

    Energy Technology Data Exchange (ETDEWEB)

    Hoop, R.C.; Schwartz, L.S.; Hoffman, E.P. [Univ. of Pittsburgh School of Medicine, Pittsburgh, PA (United States); Russo, L.S. [Univ. of Florida, Jacksonville, FL (United States); Riconda, D.L. [Orlando Regional Medical Center, Orlando, FL (United States)

    1994-02-01

    Two male cousins with Duchenne muscular dystrophy were found to have different maternal dystrophin gene haplotypes and different deletion mutations. One propositus showed two noncontiguous deletions-one in the 5{prime}, proximal deletional hotspot region, and the other in the 3{prime}, more distal deletional hotspot region. The second propositus showed only the 5{prime} deletion. Using multiple fluorescent exon dosage and fluorescent multiplex CA repeat linkage analyses, the authors show that the mother of each propositus carries both deletions on the same grandmaternal X chromosome. This paradox is explained by a single recombinational event between the 2 deleted regions of one of the carrier`s dystrophin genes, giving rise to a son with a partially {open_quotes}repaired{close_quotes} gene retaining only the 5{prime} deletion. 20 refs., 4 figs.

  19. Relatively low proportion of dystrophin gene deletions in Israeili Duchenne and Becker muscular dystrophy patients

    Energy Technology Data Exchange (ETDEWEB)

    Shomrat, R.; Gluck, E.; Legum, C.; Shiloh, Y. [Tel Aviv Univ. (Israel)

    1994-02-15

    Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations in the X-linked dystrophin gene. The most common mutations in western populations are deletions that are spread non-randomly throughout the gene. Molecular analysis of the dystrophin gene structure by hybridization of the full length cDNA to Southern blots and by PCR in 62 unrelated Israeli male DMD/BMD patients showed deletions in 23 (37%). This proportion is significantly lower than that found in European and North American populations (55-65%). Seventy-eight percent of the deletions were confined to exons 44-52, half of these exons 44-45, and the remaining 22% to exons 1 and 19. There was no correlation between the size of the deletion and the severity of the disease. All the deletions causing frameshift resulted in the DMD phenotypes. 43 refs., 1 fig., 1 tab.

  20. Context Dependent Effects of Chimeric Peptide Morpholino Conjugates Contribute to Dystrophin Exon-skipping Efficiency

    OpenAIRE

    HaiFang Yin; Prisca Boisguerin; Hong M Moulton; Corinne Betts; Yiqi Seow; Jordan Boutilier; Qingsong Wang; Anthony Walsh; Bernard Lebleu; Matthew JA Wood

    2013-01-01

    We have recently reported that cell-penetrating peptides (CPPs) and novel chimeric peptides containing CPP (referred as B peptide) and muscle-targeting peptide (referred as MSP) motifs significantly improve the systemic exon-skipping activity of morpholino phosphorodiamidate oligomers (PMOs) in dystrophin-deficient mdx mice. In the present study, the general mechanistic significance of the chimeric peptide configuration on the activity and tissue uptake of peptide conjugated PMOs in vivo was ...

  1. Ex vivo stretch reveals altered mechanical properties of isolated dystrophin-deficient hearts.

    Directory of Open Access Journals (Sweden)

    Matthew S Barnabei

    Full Text Available Duchenne muscular dystrophy (DMD is a progressive and fatal disease of muscle wasting caused by loss of the cytoskeletal protein dystrophin. In the heart, DMD results in progressive cardiomyopathy and dilation of the left ventricle through mechanisms that are not fully understood. Previous reports have shown that loss of dystrophin causes sarcolemmal instability and reduced mechanical compliance of isolated cardiac myocytes. To expand upon these findings, here we have subjected the left ventricles of dystrophin-deficient mdx hearts to mechanical stretch. Unexpectedly, isolated mdx hearts showed increased left ventricular (LV compliance compared to controls during stretch as LV volume was increased above normal end diastolic volume. During LV chamber distention, sarcomere lengths increased similarly in mdx and WT hearts despite greater excursions in volume of mdx hearts. This suggests that the mechanical properties of the intact heart cannot be modeled as a simple extrapolation of findings in single cardiac myocytes. To explain these findings, a model is proposed in which disruption of the dystrophin-glycoprotein complex perturbs cell-extracellular matrix contacts and promotes the apparent slippage of myocytes past each other during LV distension. In comparison, similar increases in LV compliance were obtained in isolated hearts from β-sarcoglycan-null and laminin-α(2 mutant mice, but not in dysferlin-null mice, suggesting that increased whole-organ compliance in mdx mice is a specific effect of disrupted cell-extracellular matrix contacts and not a general consequence of cardiomyopathy via membrane defect processes. Collectively, these findings suggest a novel and cell-death independent mechanism for the progressive pathological LV dilation that occurs in DMD.

  2. Intellectual Ability in the Duchenne Muscular Dystrophy and Dystrophin Gene Mutation Location

    Directory of Open Access Journals (Sweden)

    Rasic Milic V.

    2014-12-01

    Full Text Available Duchenne muscular dystrophy (DMD is the most common form of muscular dystrophy during childhood. Mutations in dystrophin (DMD gene are also recognized as a cause of cognitive impairment. We aimed to determine the association between intelligence level and mutation location in DMD genes in Serbian patients with DMD. Forty-one male patients with DMD, aged 3 to 16 years, were recruited at the Clinic for Neurology and Psychiatry for Children and Youth in Belgrade, Serbia. All patients had defined DMD gene deletions or duplications [multiplex ligation- dependent probe amplification (MLPA, polymerase chain reaction (PCR] and cognitive status assessment (Wechsler Intelligence Scale for Children, Brunet-Lezine scale, Vineland-Doll scale. In 37 patients with an estimated full scale intelligence quotient (FSIQ, six (16.22% had borderline intelligence (70dystrophin isoforms and when mutations in the 5’-untranslated region (5’UTR of Dp140 (exons 45-50 were assigned to affect only Dp427 and Dp260. Mutations affecting Dp140 and Dp71/Dp40 have been associated with more frequent and more severe cognitive impairment. Finally, the same classification of mutations explained the greater proportion of FSIQ variability associated with cumulative loss of dystrophin isoforms. In conclusion, cumulative loss of dystrophin isoforms increases the risk of intellectual impairment in DMD and characterizing the genotype can define necessity of early cognitive interventions in DMD patients.

  3. Multiple species comparison of cardiac troponin T and dystrophin: unravelling the DNA behind dilated cardiomyopathy

    OpenAIRE

    England, Jennifer; Loughna, Siobhan; Rutland, Catrin S.

    2017-01-01

    Animals have frequently been used as models for human disorders and mutations. Following advances in genetic testing and treatment options, and the decreasing cost of these technologies in the clinic, mutations in both companion and commercial animals are now being investigated. A recent review highlighted the genes associated with both human and non-human dilated cardiomyopathy. Cardiac troponin T and dystrophin were observed to be associated with both human and turkey (troponin T) and canin...

  4. Haplotypes in the Dystrophin DNA Segment Point to a Mosaic Origin of Modern Human Diversity

    OpenAIRE

    Ziętkiewicz, Ewa; Yotova, Vania; Gehl, Dominik; Wambach, Tina; Arrieta, Isabel; Batzer, Mark; Cole, David E.C.; Hechtman, Peter; Kaplan, Feige; Modiano, David; Moisan, Jean-Paul; Michalski, Roman; Labuda, Damian

    2003-01-01

    Although Africa has played a central role in human evolutionary history, certain studies have suggested that not all contemporary human genetic diversity is of recent African origin. We investigated 35 simple polymorphic sites and one Tn microsatellite in an 8-kb segment of the dystrophin gene. We found 86 haplotypes in 1,343 chromosomes from around the world. Although a classical out-of-Africa topology was observed in trees based on the variant frequencies, the tree of haplotype sequences re...

  5. Becker muscular dystrophy due to an intronic splicing mutation inducing a dual dystrophin transcript.

    Science.gov (United States)

    Todeschini, Alice; Gualandi, Francesca; Trabanelli, Cecilia; Armaroli, Annarita; Ravani, Anna; Fanin, Marina; Rota, Silvia; Bello, Luca; Ferlini, Alessandra; Pegoraro, Elena; Padovani, Alessandro; Filosto, Massimiliano

    2016-10-01

    We describe a 29-year-old patient who complained of left thigh muscle weakness since he was 23 and of moderate proximal weakness of both lower limbs with difficulty in climbing stairs and running since he was 27. Mild weakness of iliopsoas and quadriceps muscles and muscle atrophy of both the distal forearm and thigh were observed upon clinical examination. He harboured a novel c.1150-3C>G substitution in the DMD gene, affecting the intron 10 acceptor splice site and causing exon 11 skipping and an out-of-frame transcript. However, protein of normal molecular weight but in reduced amounts was observed on Western Blot analysis. Reverse transcription analysis on muscle RNA showed production, via alternative splicing, of a transcript missing exon 11 as well as a low abundant full-length transcript which is enough to avoid the severe Duchenne phenotype. Our study showed that a reduced amount of full length dystrophin leads to a mild form of Becker muscular dystrophy. These results confirm earlier findings that low amounts of dystrophin can be associated with a milder phenotype, which is promising for therapies aiming at dystrophin restoration. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Screening of Dystrophin Gene Deletions in Egyptian Patients with DMD/BMD Muscular Dystrophies

    Directory of Open Access Journals (Sweden)

    Laila K. Effat

    2000-01-01

    Full Text Available Duchenne muscular dystrophy (DMD and Becker muscular dystrophy (BMD are allelic disorders caused by mutations within the dystrophin gene. Our study has identified 100 Egyptian families collected from the Human Genetics Clinic, National Research Center, Cairo. All cases were subjected to complete clinical evaluation pedigree analysis, electromyography studies, estimation of serum creatine phosphokinase enzyme (CPK levels and DNA analysis. Multiplex PCR using 18 pairs of specific primers were used for screening of deletion mutations within the dystrophin gene. A frequency of 55% among the families. Sixty per cent of detected deletions involved multiple exons spanning the major or the minor hot spot of the dystrophin gene. The remainder 40% which mainly involved exon 45. Comparing these findings with frequencies of other countries it was found that our figures fall within the reported range of 40%– for deletions. The distribution of deletions in our study and other different studies was variable and specific ethnic differences do not apparently account for specific deletions. In addition this study concluded that employment of the 18 exon analysis is a cost effective and a highly accurate (97% to launch a nationwide program.

  7. Dual exon skipping in myostatin and dystrophin for Duchenne muscular dystrophy

    Directory of Open Access Journals (Sweden)

    van Ommen Gert Jan B

    2011-04-01

    Full Text Available Abstract Background Myostatin is a potent muscle growth inhibitor that belongs to the Transforming Growth Factor-β (TGF-β family. Mutations leading to non functional myostatin have been associated with hypermuscularity in several organisms. By contrast, Duchenne muscular dystrophy (DMD is characterized by a loss of muscle fibers and impaired regeneration. In this study, we aim to knockdown myostatin by means of exon skipping, a technique which has been successfully applied to reframe the genetic defect of dystrophin gene in DMD patients. Methods We targeted myostatin exon 2 using antisense oligonucleotides (AON in healthy and DMD-derived myotubes cultures. We assessed the exon skipping level, transcriptional expression of myostatin and its target genes, and combined myostatin and several dystrophin AONs. These AONs were also applied in the mdx mice models via intramuscular injections. Results Myostatin AON induced exon 2 skipping in cell cultures and to a lower extent in the mdx mice. It was accompanied by decrease in myostatin mRNA and enhanced MYOG and MYF5 expression. Furthermore, combination of myostatin and dystrophin AONs induced simultaneous skipping of both genes. Conclusions We conclude that two AONs can be used to target two different genes, MSTN and DMD, in a straightforward manner. Targeting multiple ligands of TGF-beta family will be more promising as adjuvant therapies for DMD.

  8. Gene therapies that restore dystrophin expression for the treatment of Duchenne muscular dystrophy

    Science.gov (United States)

    Robinson-Hamm, Jacqueline N.; Gersbach, Charles A.

    2016-01-01

    Duchenne muscular dystrophy is one of the most common inherited genetic diseases and is caused by mutations to the DMD gene that encodes the dystrophin protein. Recent advances in genome editing and gene therapy offer hope for the development of potential therapeutics. Truncated versions of the DMD gene can be delivered to the affected tissues with viral vectors and show promising results in a variety of animal models. Genome editing with the CRISPR/Cas9 system has recently been used to restore dystrophin expression by deleting one or more exons of the DMD gene in patient cells and in a mouse model that led to functional improvement of muscle strength. Exon skipping with oligonucleotides has been successful in several animal models and evaluated in multiple clinical trials. Next-generation oligonucleotide formulations offer significant promise to build on these results. All these approaches to restoring dystrophin expression are encouraging, but many hurdles remain. This review summarizes the current state of these technologies and summarizes considerations for their future development. PMID:27542949

  9. Single Cell Analysis of Dystrophin and SRY Gene by Using Whole Genome Amplification

    Institute of Scientific and Technical Information of China (English)

    徐晨明; 金帆; 黄荷凤; 陶冶; 叶英辉

    2001-01-01

    Objective To develop a reliable and sensitive method for detection of sex and multiloci of Duchenne muscular dystrophy (DMD) gene in single cell Materials & methods Whole genome of single cell were amplified by using 15-base random primers (primer extension preamplification, PEP), then a small aliquot of PEP product were analyzed by using locus-specific nest PCR amplification. The procedure was evaluated by detection dystrophin exons 8, 17, 19, 44, 45, 48 and human testis-determining gene (SRY)in single lymphocytes from known sources and single blastomeres from the couples with no family history of DMD.Results The amplification efficiency rate of six dystrophin exons from single lymphocytes and single blastomeres were 97. 2% (175/180) and 100% (60/60) respectively.Results of SRY showed that 100% (15/15) amplification in single male-derived lymphocytes and 0% (0/15) amplification in single female-derived lymphocytes. Conclusion The technique of single cell PEP-nest PCR for dystrophin exons 8, 17,19, 44, 45, 48 and SRY is highly specifc. PEP-nest PCR is suitable for Preimplantation genetic diagnosis (PGD) of DMD at single cell level.

  10. Interdependence of laminin-mediated clustering of lipid rafts and the dystrophin complex in astrocytes.

    Science.gov (United States)

    Noël, Geoffroy; Tham, Daniel Kai Long; Moukhles, Hakima

    2009-07-17

    Astrocyte endfeet surrounding blood vessels are active domains involved in water and potassium ion transport crucial to the maintenance of water and potassium ion homeostasis in brain. A growing body of evidence points to a role for dystroglycan and its interaction with perivascular laminin in the targeting of the dystrophin complex and the water-permeable channel, aquaporin 4 (AQP4), at astrocyte endfeet. However, the mechanisms underlying such compartmentalization remain poorly understood. In the present study we found that AQP4 resided in Triton X-100-insoluble fraction, whereas dystroglycan was recovered in the soluble fraction in astrocytes. Cholesterol depletion resulted in the translocation of a pool of AQP4 to the soluble fraction indicating that its distribution is indeed associated with cholesterol-rich membrane domains. Upon laminin treatment AQP4 and the dystrophin complex, including dystroglycan, reorganized into laminin-associated clusters enriched for the lipid raft markers GM1 and flotillin-1 but not caveolin-1. Reduced diffusion rates of GM1 in the laminin-induced clusters were indicative of the reorganization of raft components in these domains. In addition, both cholesterol depletion and dystroglycan silencing reduced the number and area of laminin-induced clusters of GM1, AQP4, and dystroglycan. These findings demonstrate the interdependence between laminin binding to dystroglycan and GM1-containing lipid raft reorganization and provide novel insight into the dystrophin complex regulation of AQP4 polarization in astrocytes.

  11. Immobilization and therapeutic passive stretching generate thickening and increase the expression of laminin and dystrophin in skeletal muscle

    International Nuclear Information System (INIS)

    Cação-Benedini, L.O.; Ribeiro, P.G.; Prado, C.M.; Chesca, D.L.; Mattiello-Sverzut, A.C.

    2014-01-01

    Extracellular matrix and costamere proteins transmit the concentric, isometric, and eccentric forces produced by active muscle contraction. The expression of these proteins after application of passive tension stimuli to muscle remains unknown. This study investigated the expression of laminin and dystrophin in the soleus muscle of rats immobilized with the right ankle in plantar flexion for 10 days and subsequent remobilization, either by isolated free movement in a cage or associated with passive stretching for up to 10 days. The intensity of the macrophage response was also evaluated. One hundred and twenty-eight female Wistar rats were divided into 8 groups: free for 10 days; immobilized for 10 days; immobilized/free for 1, 3, or 10 days; or immobilized/stretched/free for 1, 3, or 10 days. After the experimental procedures, muscle tissue was processed for immunofluorescence (dystrophin/laminin/CD68) and Western blot analysis (dystrophin/laminin). Immobilization increased the expression of dystrophin and laminin but did not alter the number of macrophages in the muscle. In the stretched muscle groups, there was an increase in dystrophin and the number of macrophages after 3 days compared with the other groups; dystrophin showed a discontinuous labeling pattern, and laminin was found in the intracellular space. The amount of laminin was increased in the muscles treated by immobilization followed by free movement for 10 days. In the initial stages of postimmobilization (1 and 3 days), an exacerbated macrophage response and an increase of dystrophin suggested that the therapeutic stretching technique induced additional stress in the muscle fibers and costameres

  12. Immobilization and therapeutic passive stretching generate thickening and increase the expression of laminin and dystrophin in skeletal muscle

    Energy Technology Data Exchange (ETDEWEB)

    Cação-Benedini, L.O.; Ribeiro, P.G. [Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, Medicina e Reabilitação do Aparelho Locomotor, Departamento de Biomecânica, Ribeirão Preto, SP, Brasil, Departamento de Biomecânica, Medicina e Reabilitação do Aparelho Locomotor, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Prado, C.M.; Chesca, D.L. [Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, Departamento de Patologia, Ribeirão Preto, SP, Brasil, Departamento de Patologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Mattiello-Sverzut, A.C. [Universidade de São Paulo, Faculdade de Medicina de Ribeirão Preto, Medicina e Reabilitação do Aparelho Locomotor, Departamento de Biomecânica, Ribeirão Preto, SP, Brasil, Departamento de Biomecânica, Medicina e Reabilitação do Aparelho Locomotor, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil)

    2014-05-09

    Extracellular matrix and costamere proteins transmit the concentric, isometric, and eccentric forces produced by active muscle contraction. The expression of these proteins after application of passive tension stimuli to muscle remains unknown. This study investigated the expression of laminin and dystrophin in the soleus muscle of rats immobilized with the right ankle in plantar flexion for 10 days and subsequent remobilization, either by isolated free movement in a cage or associated with passive stretching for up to 10 days. The intensity of the macrophage response was also evaluated. One hundred and twenty-eight female Wistar rats were divided into 8 groups: free for 10 days; immobilized for 10 days; immobilized/free for 1, 3, or 10 days; or immobilized/stretched/free for 1, 3, or 10 days. After the experimental procedures, muscle tissue was processed for immunofluorescence (dystrophin/laminin/CD68) and Western blot analysis (dystrophin/laminin). Immobilization increased the expression of dystrophin and laminin but did not alter the number of macrophages in the muscle. In the stretched muscle groups, there was an increase in dystrophin and the number of macrophages after 3 days compared with the other groups; dystrophin showed a discontinuous labeling pattern, and laminin was found in the intracellular space. The amount of laminin was increased in the muscles treated by immobilization followed by free movement for 10 days. In the initial stages of postimmobilization (1 and 3 days), an exacerbated macrophage response and an increase of dystrophin suggested that the therapeutic stretching technique induced additional stress in the muscle fibers and costameres.

  13. Alterations in the muscle force transfer apparatus in aged rats during unloading and reloading: Impact of microRNA-31.

    Science.gov (United States)

    Hughes, David C; Marcotte, George R; Baehr, Leslie M; West, Daniel W D; Marshall, Andrea G; Ebert, Scott M; Davidyan, Arik; Adams, Christopher M; Bodine, Sue C; Baar, Keith

    2018-05-03

    Force transfer is integral for maintaining skeletal muscle structure and function. One important component is dystrophin. There is limited understanding of how force transfer is impacted by age and loading. Here, we investigate the force transfer apparatus in muscles of adult and old rats exposed to periods of disuse and reloading. Our results demonstrate an increase in dystrophin protein during the reloading phase in the adult TA muscle that is delayed in old. The consequence of this delay is an increased susceptibility towards contraction-induced muscle injury. Central to the lack of dystrophin protein is an increase in miR-31, a microRNA that inhibits dystrophin translation. In vivo electroporation with a miR-31 sponge led to increased dystrophin protein and decreased contraction-induced muscle injury in old skeletal muscle. Overall, our results detail the importance of the force transfer apparatus and provide new mechanisms for contraction-induced injury in aging skeletal muscle. In healthy muscle, the dystrophin-associated glycoprotein (DGC) and integrin/focal adhesion complexes, intermediate filaments, and Z-line proteins transmit force from the contractile proteins to the extracellular matrix. How loading and age affect these proteins is poorly understood. The experiments reported here sought to determine the effect of aging on the force transfer apparatus following muscle unloading and reloading. Adult (9 months) and old (29 months) rats were subjected to 14 days hindlimb unloading (HU) and 1, 3, 7 and 14 days of reloading (REL). The DGC complex, intermediate filament and z-line protein and mRNA levels, as well as dystrophin-targeting miRNAs (miR-31, -146b and -374) were examined in the tibialis anterior (TA) and medial gastrocnemius (MG) muscles at both ages. There was a significant increase in dystrophin protein levels (2.79-fold) upon 3 days of reloading in the adult TA muscle that did not occur in the old rats (p ≤ 0.05), and the rise in

  14. Characterization of genetic deletions in Becker muscular dystrophy using monoclonal antibodies against a deletion-prone region of dystrophin

    Energy Technology Data Exchange (ETDEWEB)

    Thanh, L.T.; Man, Nguyen Thi; Morris, G.E. [Wales Institute, Clwyd (United Kingdom)] [and others

    1995-08-28

    We have produced a new panel of 20 monoclonal antibodies (mAbs) against a region of the dystrophin protein corresponding to a deletion-prone region of the Duchenne muscular dystrophy gene (exons 45-50). We show that immunohistochemistry or Western blotting with these {open_quotes}exon-specific{close_quotes} mAbs can provide a valuable addition to Southern blotting or PCR methods for the accurate identification of genetic deletions in Becker muscular dystrophy patients. The antibodies were mapped to the following exons: exon 45 (2 mAbs), exon 46 (6), exon 47 (1), exons 47/48 (4), exons 48-50 (6), and exon 50 (1). PCR amplification of single exons or groups of exons was used both to produce specific dystrophin immunogens and to map the mAbs obtained. PCR-mediated mutagenesis was also used to identify regions of dystrophin important for mAb binding. Because the mAbs can be used to characterize the dystrophin produced by individual muscle fibres, they will also be useful for studying {open_quotes}revertant{close_quotes} fibres in Duchenne muscle and for monitoring the results of myoblast therapy trials in MD patients with deletions in this region of the dystrophin gene. 27 refs., 7 figs., 3 tabs.

  15. Cognitive dysfunction in the dystrophin-deficient mouse model of Duchenne muscular dystrophy: A reappraisal from sensory to executive processes.

    Science.gov (United States)

    Chaussenot, Rémi; Edeline, Jean-Marc; Le Bec, Benoit; El Massioui, Nicole; Laroche, Serge; Vaillend, Cyrille

    2015-10-01

    Duchenne muscular dystrophy (DMD) is associated with language disabilities and deficits in learning and memory, leading to intellectual disability in a patient subpopulation. Recent studies suggest the presence of broader deficits affecting information processing, short-term memory and executive functions. While the absence of the full-length dystrophin (Dp427) is a common feature in all patients, variable mutation profiles may additionally alter distinct dystrophin-gene products encoded by separate promoters. However, the nature of the cognitive dysfunctions specifically associated with the loss of distinct brain dystrophins is unclear. Here we show that the loss of the full-length brain dystrophin in mdx mice does not modify the perception and sensorimotor gating of auditory inputs, as assessed using auditory brainstem recordings and prepulse inhibition of startle reflex. In contrast, both acquisition and long-term retention of cued and trace fear memories were impaired in mdx mice, suggesting alteration in a functional circuit including the amygdala. Spatial learning in the water maze revealed reduced path efficiency, suggesting qualitative alteration in mdx mice learning strategy. However, spatial working memory performance and cognitive flexibility challenged in various behavioral paradigms in water and radial-arm mazes were unimpaired. The full-length brain dystrophin therefore appears to play a role during acquisition of associative learning as well as in general processes involved in memory consolidation, but no overt involvement in working memory and/or executive functions could be demonstrated in spatial learning tasks. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. A case report: Becker muscular dystrophy presenting with epilepsy and dysgnosia induced by duplication mutation of Dystrophin gene.

    Science.gov (United States)

    Miao, Jing; Feng, Jia-Chun; Zhu, Dan; Yu, Xue-Fan

    2016-12-12

    Becker muscular dystrophy (BMD), a genetic disorder of X-linked recessive inheritance, typically presents with gradually progressive muscle weakness. The condition is caused by mutations of Dystrophin gene located at Xp21.2. Epilepsy is an infrequent manifestation of BMD, while cases of BMD with dysgnosia are extremely rare. We describe a 9-year-old boy with BMD, who presented with epilepsy and dysgnosia. Serum creatine kinase level was markedly elevated (3665 U/L). Wechsler intelligence tests showed a low intelligence quotient (IQ = 65). Electromyogram showed slight myogenic changes and skeletal muscle biopsy revealed muscular dystrophy. Immunohistochemical staining showed partial positivity of sarcolemma for dystrophin-N. Multiplex ligation-dependent probe amplification revealed a duplication mutation in exons 37-44 in the Dystrophin gene. The present case report helps to better understand the clinical and genetic features of BMD.

  17. Screening of point mutations by multiple SSCP analysis in the dystrophin gene

    Energy Technology Data Exchange (ETDEWEB)

    Lasa, A.; Baiget, M.; Gallano, P. [Hospital Sant Pau, Barcelona (Spain)

    1994-09-01

    Duchenne muscular dystrophy (DMD) is a lethal, X-linked neuromuscular disorder. The population frequency of DMD is one in approximately 3500 boys, of which one third is thought to be a new mutant. The DMD gene is the largest known to date, spanning over 2,3 Mb in band Xp21.2; 79 exons are transcribed into a 14 Kb mRNA coding for a protein of 427 kD which has been named dystrophin. It has been shown that about 65% of affected boys have a gene deletion with a wide variation in localization and size. The remaining affected individuals who have no detectable deletions or duplications would probably carry more subtle mutations that are difficult to detect. These mutations occur in several different exons and seem to be unique to single patients. Their identification represents a formidable goal because of the large size and complexity of the dystrophin gene. SSCP is a very efficient method for the detection of point mutations if the parameters that affect the separation of the strands are optimized for a particular DNA fragment. The multiple SSCP allows the simultaneous study of several exons, and implies the use of different conditions because no single set of conditions will be optimal for all fragments. Seventy-eight DMD patients with no deletion or duplication in the dystrophin gene were selected for the multiple SSCP analysis. Genomic DNA from these patients was amplified using the primers described for the diagnosis procedure (muscle promoter and exons 3, 8, 12, 16, 17, 19, 32, 45, 48 and 51). We have observed different mobility shifts in bands corresponding to exons 8, 12, 43 and 51. In exons 17 and 45, altered electrophoretic patterns were found in different samples identifying polymorphisms already described.

  18. Deletion of Dystrophin In-Frame Exon 5 Leads to a Severe Phenotype: Guidance for Exon Skipping Strategies.

    Directory of Open Access Journals (Sweden)

    Zhi Yon Charles Toh

    Full Text Available Duchenne and Becker muscular dystrophy severity depends upon the nature and location of the DMD gene lesion and generally correlates with the dystrophin open reading frame. However, there are striking exceptions where an in-frame genomic deletion leads to severe pathology or protein-truncating mutations (nonsense or frame-shifting indels manifest as mild disease. Exceptions to the dystrophin reading frame rule are usually resolved after molecular diagnosis on muscle RNA. We report a moderate/severe Becker muscular dystrophy patient with an in-frame genomic deletion of DMD exon 5. This mutation has been reported by others as resulting in Duchenne or Intermediate muscular dystrophy, and the loss of this in-frame exon in one patient led to multiple splicing events, including omission of exon 6, that disrupts the open reading frame and is consistent with a severe phenotype. The patient described has a deletion of dystrophin exon 5 that does not compromise recognition of exon 6, and although the deletion does not disrupt the reading frame, his clinical presentation is more severe than would be expected for classical Becker muscular dystrophy. We suggest that the dystrophin isoform lacking the actin-binding sequence encoded by exon 5 is compromised, reflected by the phenotype resulting from induction of this dystrophin isoform in mouse muscle in vivo. Hence, exon skipping to address DMD-causing mutations within DMD exon 5 may not yield an isoform that confers marked clinical benefit. Additional studies will be required to determine whether multi-exon skipping strategies could yield more functional dystrophin isoforms, since some BMD patients with larger in-frame deletions in this region have been reported with mild phenotypes.

  19. Creation of Dystrophin Expressing Chimeric Cells of Myoblast Origin as a Novel Stem Cell Based Therapy for Duchenne Muscular Dystrophy.

    Science.gov (United States)

    Siemionow, M; Cwykiel, J; Heydemann, A; Garcia-Martinez, J; Siemionow, K; Szilagyi, E

    2018-04-01

    Over the past decade different stem cell (SC) based approaches were tested to treat Duchenne Muscular Dystrophy (DMD), a lethal X-linked disorder caused by mutations in dystrophin gene. Despite research efforts, there is no curative therapy for DMD. Allogeneic SC therapies aim to restore dystrophin in the affected muscles; however, they are challenged by rejection and limited engraftment. Thus, there is a need to develop new more efficacious SC therapies. Chimeric Cells (CC), created via ex vivo fusion of donor and recipient cells, represent a promising therapeutic option for tissue regeneration and Vascularized Composite Allotransplantation (VCA) due to tolerogenic properties that eliminate the need for lifelong immunosuppression. This proof of concept study tested feasibility of myoblast fusion for Dystrophin Expressing. Chimeric Cell (DEC) therapy through in vitro characterization and in vivo assessment of engraftment, survival, and efficacy in the mdx mouse model of DMD. Murine DEC were created via ex vivo fusion of normal (snj) and dystrophin-deficient (mdx) myoblasts using polyethylene glycol. Efficacy of myoblast fusion was confirmed by flow cytometry and dystrophin immunostaining, while proliferative and myogenic differentiation capacity of DEC were assessed in vitro. Therapeutic effect after DEC transplant (0.5 × 10 6 ) into the gastrocnemius muscle (GM) of mdx mice was assessed by muscle functional tests. At 30 days post-transplant dystrophin expression in GM of injected mdx mice increased to 37.27 ± 12.1% and correlated with improvement of muscle strength and function. Our study confirmed feasibility and efficacy of DEC therapy and represents a novel SC based approach for treatment of muscular dystrophies.

  20. A sensitive, reproducible and objective immunofluorescence analysis method of dystrophin in individual fibers in samples from patients with duchenne muscular dystrophy.

    Directory of Open Access Journals (Sweden)

    Chantal Beekman

    Full Text Available Duchenne muscular dystrophy (DMD is characterized by the absence or reduced levels of dystrophin expression on the inner surface of the sarcolemmal membrane of muscle fibers. Clinical development of therapeutic approaches aiming to increase dystrophin levels requires sensitive and reproducible measurement of differences in dystrophin expression in muscle biopsies of treated patients with DMD. This, however, poses a technical challenge due to intra- and inter-donor variance in the occurrence of revertant fibers and low trace dystrophin expression throughout the biopsies. We have developed an immunofluorescence and semi-automated image analysis method that measures the sarcolemmal dystrophin intensity per individual fiber for the entire fiber population in a muscle biopsy. Cross-sections of muscle co-stained for dystrophin and spectrin have been imaged by confocal microscopy, and image analysis was performed using Definiens software. Dystrophin intensity has been measured in the sarcolemmal mask of spectrin for each individual muscle fiber and multiple membrane intensity parameters (mean, maximum, quantiles per fiber were calculated. A histogram can depict the distribution of dystrophin intensities for the fiber population in the biopsy. This method was tested by measuring dystrophin in DMD, Becker muscular dystrophy, and healthy muscle samples. Analysis of duplicate or quadruplicate sections of DMD biopsies on the same or multiple days, by different operators, or using different antibodies, was shown to be objective and reproducible (inter-assay precision, CV 2-17% and intra-assay precision, CV 2-10%. Moreover, the method was sufficiently sensitive to detect consistently small differences in dystrophin between two biopsies from a patient with DMD before and after treatment with an investigational compound.

  1. Becker muscular dystrophy severity is linked to the structure of dystrophin.

    Science.gov (United States)

    Nicolas, Aurélie; Raguénès-Nicol, Céline; Ben Yaou, Rabah; Ameziane-Le Hir, Sarah; Chéron, Angélique; Vié, Véronique; Claustres, Mireille; Leturcq, France; Delalande, Olivier; Hubert, Jean-François; Tuffery-Giraud, Sylvie; Giudice, Emmanuel; Le Rumeur, Elisabeth

    2015-03-01

    In-frame exon deletions of the Duchenne muscular dystrophy (DMD) gene produce internally truncated proteins that typically lead to Becker muscular dystrophy (BMD), a milder allelic disorder of DMD. We hypothesized that differences in the structure of mutant dystrophin may be responsible for the clinical heterogeneity observed in Becker patients and we studied four prevalent in-frame exon deletions, i.e. Δ45-47, Δ45-48, Δ45-49 and Δ45-51. Molecular homology modelling revealed that the proteins corresponding to deletions Δ45-48 and Δ45-51 displayed a similar structure (hybrid repeat) than the wild-type dystrophin, whereas deletions Δ45-47 and Δ45-49 lead to proteins with an unrelated structure (fractional repeat). All four proteins in vitro expressed in a fragment encoding repeats 16-21 were folded in α-helices and remained highly stable. Refolding dynamics were slowed and molecular surface hydrophobicity were higher in fractional repeat containing Δ45-47 and Δ45-49 deletions compared with hybrid repeat containing Δ45-48 and Δ45-51 deletions. By retrospectively collecting data for a series of French BMD patients, we showed that the age of dilated cardiomyopathy (DCM) onset was delayed by 11 and 14 years in Δ45-48 and Δ45-49 compared with Δ45-47 patients, respectively. A clear trend toward earlier wheelchair dependency (minimum of 11 years) was also observed in Δ45-47 and Δ45-49 patients compared with Δ45-48 patients. Muscle dystrophin levels were moderately reduced in most patients without clear correlation with the deletion type. Disease progression in BMD patients appears to be dependent on the deletion itself and associated with a specific structure of dystrophin at the deletion site. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  2. Nanopolymers improve delivery of exon skipping oligonucleotides and concomitant dystrophin expression in skeletal muscle of mdx mice

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    Sirsi Shashank R

    2008-04-01

    Full Text Available Abstract Background Exon skipping oligonucleotides (ESOs of 2'O-Methyl (2'OMe and morpholino chemistry have been shown to restore dystrophin expression in muscle fibers from the mdx mouse, and are currently being tested in phase I clinical trials for Duchenne Muscular Dystrophy (DMD. However, ESOs remain limited in their effectiveness because of an inadequate delivery profile. Synthetic cationic copolymers of poly(ethylene imine (PEI and poly(ethylene glycol (PEG are regarded as effective agents for enhanced delivery of nucleic acids in various applications. Results We examined whether PEG-PEI copolymers can facilitate ESO-mediated dystrophin expression after intramuscular injections into tibialis anterior (TA muscles of mdx mice. We utilized a set of PEG-PEI copolymers containing 2 kDa PEI and either 550 Da or 5 kDa PEG, both of which bind 2'OMe ESOs with high affinity and form stable nanoparticulates with a relatively low surface charge. Three weekly intramuscular injections of 5 μg of ESO complexed with PEI2K-PEG550 copolymers resulted in about 500 dystrophin-positive fibers and about 12% of normal levels of dystrophin expression at 3 weeks after the initial injection, which is significantly greater than for injections of ESO alone, which are known to be almost completely ineffective. In an effort to enhance biocompatibility and cellular uptake, the PEI2K-PEG550 and PEI2K-PEG5K copolymers were functionalized by covalent conjugation with nanogold (NG or adsorbtion of colloidal gold (CG, respectively. Surprisingly, using the same injection and dosing regimen, we found no significant difference in dystrophin expression by Western blot between the NG-PEI2K-PEG550, CG-PEI2K-PEG5K, and non-functionalized PEI2K-PEG550 copolymers. Dose-response experiments using the CG-PEI2K-PEG5K copolymer with total ESO ranging from 3–60 μg yielded a maximum of about 15% dystrophin expression. Further improvements in dystrophin expression up to 20% of normal

  3. Nonmechanical Roles of Dystrophin and Associated Proteins in Exercise, Neuromuscular Junctions, and Brains

    Directory of Open Access Journals (Sweden)

    Bailey Nichols

    2015-07-01

    Full Text Available Dystrophin-glycoprotein complex (DGC is an important structural unit in skeletal muscle that connects the cytoskeleton (f-actin of a muscle fiber to the extracellular matrix (ECM. Several muscular dystrophies, such as Duchenne muscular dystrophy, Becker muscular dystrophy, congenital muscular dystrophies (dystroglycanopathies, and limb-girdle muscular dystrophies (sarcoglycanopathies, are caused by mutations in the different DGC components. Although many early studies indicated DGC plays a crucial mechanical role in maintaining the structural integrity of skeletal muscle, recent studies identified novel roles of DGC. Beyond a mechanical role, these DGC members play important signaling roles and act as a scaffold for various signaling pathways. For example, neuronal nitric oxide synthase (nNOS, which is localized at the muscle membrane by DGC members (dystrophin and syntrophins, plays an important role in the regulation of the blood flow during exercise. DGC also plays important roles at the neuromuscular junction (NMJ and in the brain. In this review, we will focus on recently identified roles of DGC particularly in exercise and the brain.

  4. Fetal skeletal muscle progenitors have regenerative capacity after intramuscular engraftment in dystrophin deficient mice.

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    Hiroshi Sakai

    Full Text Available Muscle satellite cells (SCs are stem cells that reside in skeletal muscles and contribute to regeneration upon muscle injury. SCs arise from skeletal muscle progenitors expressing transcription factors Pax3 and/or Pax7 during embryogenesis in mice. However, it is unclear whether these fetal progenitors possess regenerative ability when transplanted in adult muscle. Here we address this question by investigating whether fetal skeletal muscle progenitors (FMPs isolated from Pax3(GFP/+ embryos have the capacity to regenerate muscle after engraftment into Dystrophin-deficient mice, a model of Duchenne muscular dystrophy. The capacity of FMPs to engraft and enter the myogenic program in regenerating muscle was compared with that of SCs derived from adult Pax3(GFP/+ mice. Transplanted FMPs contributed to the reconstitution of damaged myofibers in Dystrophin-deficient mice. However, despite FMPs and SCs having similar myogenic ability in culture, the regenerative ability of FMPs was less than that of SCs in vivo. FMPs that had activated MyoD engrafted more efficiently to regenerate myofibers than MyoD-negative FMPs. Transcriptome and surface marker analyses of these cells suggest the importance of myogenic priming for the efficient myogenic engraftment. Our findings suggest the regenerative capability of FMPs in the context of muscle repair and cell therapy for degenerative muscle disease.

  5. Characterization of dystrophin deficient rats: a new model for Duchenne muscular dystrophy.

    Science.gov (United States)

    Larcher, Thibaut; Lafoux, Aude; Tesson, Laurent; Remy, Séverine; Thepenier, Virginie; François, Virginie; Le Guiner, Caroline; Goubin, Helicia; Dutilleul, Maéva; Guigand, Lydie; Toumaniantz, Gilles; De Cian, Anne; Boix, Charlotte; Renaud, Jean-Baptiste; Cherel, Yan; Giovannangeli, Carine; Concordet, Jean-Paul; Anegon, Ignacio; Huchet, Corinne

    2014-01-01

    A few animal models of Duchenne muscular dystrophy (DMD) are available, large ones such as pigs or dogs being expensive and difficult to handle. Mdx (X-linked muscular dystrophy) mice only partially mimic the human disease, with limited chronic muscular lesions and muscle weakness. Their small size also imposes limitations on analyses. A rat model could represent a useful alternative since rats are small animals but 10 times bigger than mice and could better reflect the lesions and functional abnormalities observed in DMD patients. Two lines of Dmd mutated-rats (Dmdmdx) were generated using TALENs targeting exon 23. Muscles of animals of both lines showed undetectable levels of dystrophin by western blot and less than 5% of dystrophin positive fibers by immunohistochemistry. At 3 months, limb and diaphragm muscles from Dmdmdx rats displayed severe necrosis and regeneration. At 7 months, these muscles also showed severe fibrosis and some adipose tissue infiltration. Dmdmdx rats showed significant reduction in muscle strength and a decrease in spontaneous motor activity. Furthermore, heart morphology was indicative of dilated cardiomyopathy associated histologically with necrotic and fibrotic changes. Echocardiography showed significant concentric remodeling and alteration of diastolic function. In conclusion, Dmdmdx rats represent a new faithful small animal model of DMD.

  6. Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle

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    Camilla Brolin

    2015-01-01

    Full Text Available Peptide nucleic acid (PNA is a synthetic DNA mimic that has shown potential for discovery of novel splice switching antisense drugs. However, in vivo cellular delivery has been a limiting factor for development, and only few successful studies have been reported. As a possible modality for improvement of in vivo cellular availability, we have investigated the effect of electrotransfer upon intramuscular (i.m. PNA administration in vivo. Antisense PNA targeting exon 23 of the murine dystrophin gene was administered by i.m. injection to the tibialis anterior (TA muscle of normal NMRI and dystrophic mdx mice with or without electroporation. At low, single PNA doses (1.5, 3, or 10 µg/TA, electroporation augmented the antisense exon skipping induced by an unmodified PNA by twofold to fourfold in healthy mouse muscle with optimized electric parameters, measured after 7 days. The PNA splice switching was detected at the RNA level up to 4 weeks after a single-dose treatment. In dystrophic muscles of the MDX mouse, electroporation increased the number of dystrophin-positive fibers about 2.5-fold at 2 weeks after a single PNA administration compared to injection only. In conclusion, we find that electroporation can enhance PNA antisense effects in muscle tissue.

  7. Dystrophin-deficient cardiomyocytes derived from human urine: New biologic reagents for drug discovery

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    Xuan Guan

    2014-03-01

    Full Text Available The ability to extract somatic cells from a patient and reprogram them to pluripotency opens up new possibilities for personalized medicine. Induced pluripotent stem cells (iPSCs have been employed to generate beating cardiomyocytes from a patient's skin or blood cells. Here, iPSC methods were used to generate cardiomyocytes starting from the urine of a patient with Duchenne muscular dystrophy (DMD. Urine was chosen as a starting material because it contains adult stem cells called urine-derived stem cells (USCs. USCs express the canonical reprogramming factors c-myc and klf4, and possess high telomerase activity. Pluripotency of urine-derived iPSC clones was confirmed by immunocytochemistry, RT-PCR and teratoma formation. Urine-derived iPSC clones generated from healthy volunteers and a DMD patient were differentiated into beating cardiomyocytes using a series of small molecules in monolayer culture. Results indicate that cardiomyocytes retain the DMD patient's dystrophin mutation. Physiological assays suggest that dystrophin-deficient cardiomyocytes possess phenotypic differences from normal cardiomyocytes. These results demonstrate the feasibility of generating cardiomyocytes from a urine sample and that urine-derived cardiomyocytes retain characteristic features that might be further exploited for mechanistic studies and drug discovery.

  8. Adhalin, the 50 kD dystrophin associated protein, is not the locus for severe childhood autosomal recessive dystrophy (SCARMD)

    Energy Technology Data Exchange (ETDEWEB)

    McNally, E.M.; Selig, S.; Kunkel, L.M. [Children`s Hospital, Boston, MA (United States)

    1994-09-01

    Mutations in the carboxyl-terminus in dystrophin are normally sufficient to produce severely dystrophic muscle. This portion of dystrophin binds a complex of dystrophin-associated glycoproteins (DAGs). The genes encoding these DAGs are candidate genes for causing neuromuscular disease. Immunoreactivity for adhalin, the 50 kD DAG, is absent in muscle biopsies from patients with SCARMD, a form of dystrophy clinically similar Duchenne muscular dystrophy. Prior linkage analysis in SCARMD families revealed that the disease gene segregates with markers on chromosome 13. To determine the molecular role that adhalin may play in SCARMD, human cDNA and genomic sequences were isolated. Primers were designed based on predicted areas of conservation in rabbit adhalin and used in RT-PCR with human skeletal and cardiac muscle. RT-PCR products were confirmed by sequence as human adhalin and then used as probes for screening human cDNA and genomic libraries. Human and rabbit adhalin are 90% identical, and among the cDNAs, a novel splice form of adhalin was seen which may encode part of the 35 kD component of the dystrophin-glycoprotein complex. To our surprise, only human/rodent hybrids containing human chromosome 17 amplified adhalin sequences in a PCR analysis. FISH analysis with three overlapping genomic sequences confirmed the chromosome 17 location and further delineated the map position to 17q21. Therefore, adhalin is excluded as the gene causing SCARMD.

  9. Dual AAV Gene Therapy for Duchenne Muscular Dystrophy with a 7-kb Mini-Dystrophin Gene in the Canine Model.

    Science.gov (United States)

    Kodippili, Kasun; Hakim, Chady H; Pan, Xiufang; Yang, Hsiao T; Yue, Yongping; Zhang, Yadong; Shin, Jin-Hong; Yang, N Nora; Duan, Dongsheng

    2018-03-01

    Dual adeno-associated virus (AAV) technology was developed in 2000 to double the packaging capacity of the AAV vector. The proof of principle has been demonstrated in various mouse models. Yet, pivotal evidence is lacking in large animal models of human diseases. Here we report expression of a 7-kb canine ΔH2-R15 mini-dystrophin gene using a pair of dual AAV vectors in the canine model of Duchenne muscular dystrophy (DMD). The ΔH2-R15 minigene is by far the most potent synthetic dystrophin gene engineered for DMD gene therapy. We packaged minigene dual vectors in Y731F tyrosine-modified AAV-9 and delivered to the extensor carpi ulnaris muscle of a 12-month-old affected dog at the dose of 2 × 10 13 viral genome particles/vector/muscle. Widespread mini-dystrophin expression was observed 2 months after gene transfer. The missing dystrophin-associated glycoprotein complex was restored. Treatment also reduced muscle degeneration and fibrosis and improved myofiber size distribution. Importantly, dual AAV therapy greatly protected the muscle from eccentric contraction-induced force loss. Our data provide the first clear evidence that dual AAV therapy can be translated to a diseased large mammal. Further development of dual AAV technology may lead to effective therapies for DMD and many other diseases in human patients.

  10. Mouse models of two missense mutations in actin-binding domain 1 of dystrophin associated with Duchenne or Becker muscular dystrophy.

    Science.gov (United States)

    McCourt, Jackie L; Talsness, Dana M; Lindsay, Angus; Arpke, Robert W; Chatterton, Paul D; Nelson, D'anna M; Chamberlain, Christopher M; Olthoff, John T; Belanto, Joseph J; McCourt, Preston M; Kyba, Michael; Lowe, Dawn A; Ervasti, James M

    2018-02-01

    Missense mutations in the dystrophin protein can cause Duchenne muscular dystrophy (DMD) or Becker muscular dystrophy (BMD) through an undefined pathomechanism. In vitro studies suggest that missense mutations in the N-terminal actin-binding domain (ABD1) cause protein instability, and cultured myoblast studies reveal decreased expression levels that can be restored to wild-type with proteasome inhibitors. To further elucidate the pathophysiology of missense dystrophin in vivo, we generated two transgenic mdx mouse lines expressing L54R or L172H mutant dystrophin, which correspond to missense mutations identified in human patients with DMD or BMD, respectively. Our biochemical, histologic and physiologic analysis of the L54R and L172H mice show decreased levels of dystrophin which are proportional to the phenotypic severity. Proteasome inhibitors were ineffective in both the L54R and L172H mice, yet mice homozygous for the L172H transgene were able to express even higher levels of dystrophin which caused further improvements in muscle histology and physiology. Given that missense dystrophin is likely being degraded by the proteasome but whole body proteasome inhibition was not possible, we screened for ubiquitin-conjugating enzymes involved in targeting dystrophin to the proteasome. A myoblast cell line expressing L54R mutant dystrophin was screened with an siRNA library targeting E1, E2 and E3 ligases which identified Amn1, FBXO33, Zfand5 and Trim75. Our study establishes new mouse models of dystrophinopathy and identifies candidate E3 ligases that may specifically regulate dystrophin protein turnover in vivo. © The Author(s) 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  11. Modulation of splicing of the preceding intron by antisense oligonucleotide complementary to intra-exon sequence deleted in dystrophin Kobe

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    Takeshima, Y.; Matuso, M.; Sakamoto, H.; Nishio, H. [Kobe Univ. School of Medicine and Science (Japan)

    1994-09-01

    Molecular analysis of dystrophin Kobe showed that exon 19 of the dystrophin gene bearing a 52 bp deletion was skipped during splicing, although the known consensus sequences at the 5{prime} and 3{prime} splice site of exon 19 were maintained. These data suggest that the deleted sequence of exon 19 may function as a cis-acting factor for exact splicing for the upstream intron. To investigate this potential role, an in vitro splicing system using dystrophin precursors was established. A two-exon precursor containing exon 18, truncated intron 18, and exon 19 was accurately spliced. However, splicing of intron 18 was dramatically inhibited when wild exon 19 was replaced with mutated exon 19. Even though the length of exon 19 was restored to normal by replacing the deleted sequence with other sequence, splicing of intron 18 was not fully reactivated. Characteristically, splicing of intron 18 was inactivated more markedly when the replaced sequence contained less polypurine stretches. These data suggested that modification of the exon sequence would result in a splicing abnormality. Antisense 31 mer 2`-O-methyl ribonucleotide was targeted against 5{prime} end of deleted region of exon 19 to modulate splicing of the mRNA precursor. Splicing of intron 18 was inhibited in a dose- and time-dependent manner. This is the first in vitro evidence to show splicing of dystrophin pre-mRNA can be managed by antisense oligonucleotides. These experiments represent an approach in which antisense oligonucleotides are used to restore the function of a defective dystrophin gene in Duchenne muscular dystrophy by inducing skipping of certain exons during splicing.

  12. Phase 2a study of ataluren-mediated dystrophin production in patients with nonsense mutation Duchenne muscular dystrophy.

    Directory of Open Access Journals (Sweden)

    Richard S Finkel

    Full Text Available Approximately 13% of boys with Duchenne muscular dystrophy (DMD have a nonsense mutation in the dystrophin gene, resulting in a premature stop codon in the corresponding mRNA and failure to generate a functional protein. Ataluren (PTC124 enables ribosomal readthrough of premature stop codons, leading to production of full-length, functional proteins.This Phase 2a open-label, sequential dose-ranging trial recruited 38 boys with nonsense mutation DMD. The first cohort (n = 6 received ataluren three times per day at morning, midday, and evening doses of 4, 4, and 8 mg/kg; the second cohort (n = 20 was dosed at 10, 10, 20 mg/kg; and the third cohort (n = 12 was dosed at 20, 20, 40 mg/kg. Treatment duration was 28 days. Change in full-length dystrophin expression, as assessed by immunostaining in pre- and post-treatment muscle biopsy specimens, was the primary endpoint.Twenty three of 38 (61% subjects demonstrated increases in post-treatment dystrophin expression in a quantitative analysis assessing the ratio of dystrophin/spectrin. A qualitative analysis also showed positive changes in dystrophin expression. Expression was not associated with nonsense mutation type or exon location. Ataluren trough plasma concentrations active in the mdx mouse model were consistently achieved at the mid- and high- dose levels in participants. Ataluren was generally well tolerated.Ataluren showed activity and safety in this short-term study, supporting evaluation of ataluren 10, 10, 20 mg/kg and 20, 20, 40 mg/kg in a Phase 2b, double-blind, long-term study in nonsense mutation DMD.ClinicalTrials.gov NCT00264888.

  13. Matrix metalloproteinase-2 ablation in dystrophin-deficient mdx muscles reduces angiogenesis resulting in impaired growth of regenerated muscle fibers.

    Science.gov (United States)

    Miyazaki, Daigo; Nakamura, Akinori; Fukushima, Kazuhiro; Yoshida, Kunihiro; Takeda, Shin'ichi; Ikeda, Shu-ichi

    2011-05-01

    Matrix metalloproteases (MMPs) are a family of endopeptidases classified into subgroups based on substrate preference in normal physiological processes such as embryonic development and tissue remodeling, as well as in various disease processes via degradation of extracellular matrix components. Among the MMPs, MMP-9 and MMP-2 have been reported to be up-regulated in skeletal muscles in the lethal X-linked muscle disorder Duchenne muscular dystrophy (DMD), which is caused by loss of dystrophin. A recent study showed that deletion of the MMP9 gene in mdx, a mouse model for DMD, improved skeletal muscle pathology and function; however, the role of MMP-2 in the dystrophin-deficient muscle is not well known. In this study, we aimed at verifying the role of MMP-2 in the dystrophin-deficient muscle by using mdx mice with genetic ablation of MMP-2 (mdx/MMP-2(-/-)). We found impairment of regenerated muscle fiber growth with reduction of angiogenesis in mdx/MMP-2(-/-) mice at 3 months of age. Expression of vascular endothelial growth factor-A (VEGF-A), an important angiogenesis-related factor, decreased in mdx/MMP-2(-/-) mice at 3 months of age. MMP-2 had not a critical role in the degradation of dystrophin-glycoprotein complex (DGC) components such as β-dystroglycan and β-sarcoglycan in the regeneration process of the dystrophic muscle. Accordingly, MMP-2 may be essential for growth of regenerated muscle fibers through VEGF-associated angiogenesis in the dystrophin-deficient skeletal muscle.

  14. A BanI RFLP at a deletion hotspot in the human dystrophin gene

    Energy Technology Data Exchange (ETDEWEB)

    Read, A P; Mountford, R [St. Mary' s Hospital, Manchester (England)

    1990-01-25

    Cf56a is a 0.9 kb EcoRI fragment of dystrophin cDNA in pUC13. Cf56a is identical to Kunkel's cDNA probe 8. Constant bands of 14.4, 11.0, 8.1, 6.2 and 1.3 kb correspond to exons I, N, L, N and K respectively. The polymorphic band is exon J (exon 48, 1.2+3.9 kb HindIII bands). This exon is deleted in 25% of all Duchenne/Becker dystrophy boys. Therefore this RFLP is useful for determining carrier status of at-risk females by showing heterozygosity or apparent non-maternity.

  15. Sparks, signals and shock absorbers: how dystrophin loss causes muscular dystrophy.

    Science.gov (United States)

    Batchelor, Clare L; Winder, Steve J

    2006-04-01

    The dystrophin-glycoprotein complex (DGC) can be considered as a specialized adhesion complex, linking the extracellular matrix to the actin cytoskeleton, primarily in muscle cells. Mutations in several components of the DGC lead to its partial or total loss, resulting in various forms of muscular dystrophy. These typically manifest as progressive wasting diseases with loss of muscle integrity. Debate is ongoing about the precise function of the DGC: initially a strictly mechanical role was proposed but it has been suggested that there is aberrant calcium handling in muscular dystrophy and, more recently, changes in MAP kinase and GTPase signalling have been implicated in the aetiology of the disease. Here, we discuss new and interesting developments in these aspects of DGC function and attempt to rationalize the mechanical, calcium and signalling hypotheses to provide a unifying hypothesis of the underlying process of muscular dystrophy.

  16. A BanI RFLP at a deletion hotspot in the human dystrophin gene

    Energy Technology Data Exchange (ETDEWEB)

    Read, A.P.; Mountford, R. (St. Mary' s Hospital, Manchester (England))

    1990-01-25

    Cf56a is a 0.9 kb EcoRI fragment of dystrophin cDNA in pUC13. Cf56a is identical to Kunkel's cDNA probe 8. Constant bands of 14.4, 11.0, 8.1, 6.2 and 1.3 kb correspond to exons I, N, L, N and K respectively. The polymorphic band is exon J (exon 48, 1.2+3.9 kb HindIII bands). This exon is deleted in 25% of all Duchenne/Becker dystrophy boys. Therefore this RFLP is useful for determining carrier status of at-risk females by showing heterozygosity or apparent non-maternity.

  17. Neuronal differentiation modulates the dystrophin Dp71d binding to the nuclear matrix

    International Nuclear Information System (INIS)

    Rodriguez-Munoz, Rafael; Villarreal-Silva, Marcela; Gonzalez-Ramirez, Ricardo; Garcia-Sierra, Francisco; Mondragon, Monica; Mondragon, Ricardo; Cerna, Joel; Cisneros, Bulmaro

    2008-01-01

    The function of dystrophin Dp71 in neuronal cells remains unknown. To approach this issue, we have selected the PC12 neuronal cell line. These cells express both a Dp71f cytoplasmic variant and a Dp71d nuclear isoform. In this study, we demonstrated by electron and confocal microscopy analyses of in situ nuclear matrices and Western blotting evaluation of cell extracts that Dp71d associates with the nuclear matrix. Interestingly, this binding is modulated during NGF-induced neuronal differentiation of PC12 cells with a twofold increment in the differentiated cells, compared to control cells. Also, distribution of Dp71d along the periphery of the nuclear matrix observed in the undifferentiated cells is replaced by intense fluorescent foci localized in Center of the nucleoskeletal structure. In summary, we revealed that Dp71d is a dynamic component of nuclear matrix that might participate in the nuclear modeling occurring during neuronal differentiation

  18. Quantitative analysis of the dystrophin gene by real-time PCR

    Directory of Open Access Journals (Sweden)

    Maksimovic Nela

    2012-01-01

    Full Text Available Duchenne and Becker muscular dystrophy (DMD/BMD are severe X-linked neuromuscular disorders caused by mutations in the dystrophin gene. Our aim was to optimize a quantitative real-time PCR method based on SYBR® Green I chemistry for routine diagnostics of DMD/BMD deletion carriers. Twenty female relatives of DMD/BMD patients with previously detected partial gene deletions were studied. The relative quantity of the target exons was calculated by a comparative threshold cycle method (ΔΔCt. The carrier status of all subjects was successfully determined. The gene dosage ratio for non-carriers was 1.07±0.20, and for carriers 0.56±0.11. This assay proved to be simple, rapid, reliable and cost-effective.

  19. Context Dependent Effects of Chimeric Peptide Morpholino Conjugates Contribute to Dystrophin Exon-skipping Efficiency.

    Science.gov (United States)

    Yin, Haifang; Boisguerin, Prisca; Moulton, Hong M; Betts, Corinne; Seow, Yiqi; Boutilier, Jordan; Wang, Qingsong; Walsh, Anthony; Lebleu, Bernard; Wood, Matthew Ja

    2013-09-24

    We have recently reported that cell-penetrating peptides (CPPs) and novel chimeric peptides containing CPP (referred as B peptide) and muscle-targeting peptide (referred as MSP) motifs significantly improve the systemic exon-skipping activity of morpholino phosphorodiamidate oligomers (PMOs) in dystrophin-deficient mdx mice. In the present study, the general mechanistic significance of the chimeric peptide configuration on the activity and tissue uptake of peptide conjugated PMOs in vivo was investigated. Four additional chimeric peptide-PMO conjugates including newly identified peptide 9 (B-9-PMO and 9-B-PMO) and control peptide 3 (B-3-PMO and 3-B-PMO) were tested in mdx mice. Immunohistochemical staining, RT-PCR and western blot results indicated that B-9-PMO induced significantly higher level of exon skipping and dystrophin restoration than its counterpart (9-B-PMO), further corroborating the notion that the activity of chimeric peptide-PMO conjugates is dependent on relative position of the tissue-targeting peptide motif within the chimeric peptide with respect to PMOs. Subsequent mechanistic studies showed that enhanced cellular uptake of B-MSP-PMO into muscle cells leads to increased exon-skipping activity in comparison with MSP-B-PMO. Surprisingly, further evidence showed that the uptake of chimeric peptide-PMO conjugates of both orientations (B-MSP-PMO and MSP-B-PMO) was ATP- and temperature-dependent and also partially mediated by heparan sulfate proteoglycans (HSPG), indicating that endocytosis is likely the main uptake pathway for both chimeric peptide-PMO conjugates. Collectively, our data demonstrate that peptide orientation in chimeric peptides is an important parameter that determines cellular uptake and activity when conjugated directly to oligonucleotides. These observations provide insight into the design of improved cell targeting compounds for future therapeutics studies.Molecular Therapy-Nucleic Acids (2013) 2, e124; doi:10.1038/mtna.2013

  20. Context Dependent Effects of Chimeric Peptide Morpholino Conjugates Contribute to Dystrophin Exon-skipping Efficiency

    Directory of Open Access Journals (Sweden)

    HaiFang Yin

    2013-01-01

    Full Text Available We have recently reported that cell-penetrating peptides (CPPs and novel chimeric peptides containing CPP (referred as B peptide and muscle-targeting peptide (referred as MSP motifs significantly improve the systemic exon-skipping activity of morpholino phosphorodiamidate oligomers (PMOs in dystrophin-deficient mdx mice. In the present study, the general mechanistic significance of the chimeric peptide configuration on the activity and tissue uptake of peptide conjugated PMOs in vivo was investigated. Four additional chimeric peptide-PMO conjugates including newly identified peptide 9 (B-9-PMO and 9-B-PMO and control peptide 3 (B-3-PMO and 3-B-PMO were tested in mdx mice. Immunohistochemical staining, RT-PCR and western blot results indicated that B-9-PMO induced significantly higher level of exon skipping and dystrophin restoration than its counterpart (9-B-PMO, further corroborating the notion that the activity of chimeric peptide-PMO conjugates is dependent on relative position of the tissue-targeting peptide motif within the chimeric peptide with respect to PMOs. Subsequent mechanistic studies showed that enhanced cellular uptake of B-MSP-PMO into muscle cells leads to increased exon-skipping activity in comparison with MSP-B-PMO. Surprisingly, further evidence showed that the uptake of chimeric peptide-PMO conjugates of both orientations (B-MSP-PMO and MSP-B-PMO was ATP- and temperature-dependent and also partially mediated by heparan sulfate proteoglycans (HSPG, indicating that endocytosis is likely the main uptake pathway for both chimeric peptide-PMO conjugates. Collectively, our data demonstrate that peptide orientation in chimeric peptides is an important parameter that determines cellular uptake and activity when conjugated directly to oligonucleotides. These observations provide insight into the design of improved cell targeting compounds for future therapeutics studies.

  1. Somatodendritic and excitatory postsynaptic distribution of neuron-type dystrophin isoform, Dp40, in hippocampal neurons

    International Nuclear Information System (INIS)

    Fujimoto, Takahiro; Itoh, Kyoko; Yaoi, Takeshi; Fushiki, Shinji

    2014-01-01

    Highlights: • Identification of dystrophin (Dp) shortest isoform, Dp40, is a neuron-type Dp. • Dp40 expression is temporally and differentially regulated in comparison to Dp71. • Somatodendritic and nuclear localization of Dp40. • Dp40 is localized to excitatory postsynapses. • Dp40 might play roles in dendritic and synaptic functions. - Abstract: The Duchenne muscular dystrophy (DMD) gene produces multiple dystrophin (Dp) products due to the presence of several promoters. We previously reported the existence of a novel short isoform of Dp, Dp40, in adult mouse brain. However, the exact biochemical expression profile and cytological distribution of the Dp40 protein remain unknown. In this study, we generated a polyclonal antibody against the NH 2 -terminal region of the Dp40 and identified the expression profile of Dp40 in the mouse brain. Through an analysis using embryonic and postnatal mouse cerebrums, we found that Dp40 emerged from the early neonatal stages until adulthood, whereas Dp71, an another Dp short isoform, was highly detected in both prenatal and postnatal cerebrums. Intriguingly, relative expressions of Dp40 and Dp71 were prominent in cultured dissociated neurons and non-neuronal cells derived from mouse hippocampus, respectively. Furthermore, the immunocytological distribution of Dp40 was analyzed in dissociated cultured neurons, revealing that Dp40 is detected in the soma and its dendrites, but not in the axon. It is worthy to note that Dp40 is localized along the subplasmalemmal region of the dendritic shafts, as well as at excitatory postsynaptic sites. Thus, Dp40 was identified as a neuron-type Dp possibly involving dendritic and synaptic functions

  2. Somatodendritic and excitatory postsynaptic distribution of neuron-type dystrophin isoform, Dp40, in hippocampal neurons

    Energy Technology Data Exchange (ETDEWEB)

    Fujimoto, Takahiro; Itoh, Kyoko, E-mail: kxi14@koto.kpu-m.ac.jp; Yaoi, Takeshi; Fushiki, Shinji

    2014-09-12

    Highlights: • Identification of dystrophin (Dp) shortest isoform, Dp40, is a neuron-type Dp. • Dp40 expression is temporally and differentially regulated in comparison to Dp71. • Somatodendritic and nuclear localization of Dp40. • Dp40 is localized to excitatory postsynapses. • Dp40 might play roles in dendritic and synaptic functions. - Abstract: The Duchenne muscular dystrophy (DMD) gene produces multiple dystrophin (Dp) products due to the presence of several promoters. We previously reported the existence of a novel short isoform of Dp, Dp40, in adult mouse brain. However, the exact biochemical expression profile and cytological distribution of the Dp40 protein remain unknown. In this study, we generated a polyclonal antibody against the NH{sub 2}-terminal region of the Dp40 and identified the expression profile of Dp40 in the mouse brain. Through an analysis using embryonic and postnatal mouse cerebrums, we found that Dp40 emerged from the early neonatal stages until adulthood, whereas Dp71, an another Dp short isoform, was highly detected in both prenatal and postnatal cerebrums. Intriguingly, relative expressions of Dp40 and Dp71 were prominent in cultured dissociated neurons and non-neuronal cells derived from mouse hippocampus, respectively. Furthermore, the immunocytological distribution of Dp40 was analyzed in dissociated cultured neurons, revealing that Dp40 is detected in the soma and its dendrites, but not in the axon. It is worthy to note that Dp40 is localized along the subplasmalemmal region of the dendritic shafts, as well as at excitatory postsynaptic sites. Thus, Dp40 was identified as a neuron-type Dp possibly involving dendritic and synaptic functions.

  3. New Dystrophin/Dystroglycan interactors control neuron behavior in Drosophila eye

    Directory of Open Access Journals (Sweden)

    Rishko Valentyna M

    2011-09-01

    Full Text Available Abstract Background The Dystrophin Glycoprotein Complex (DGC is a large multi-component complex that is well known for its function in muscle tissue. When the main components of the DGC, Dystrophin (Dys and Dystroglycan (Dg are affected cognitive impairment and mental retardation in addition to muscle degeneration can occur. Previously we performed an array of genetic screens using a Drosophila model for muscular dystrophy in order to find novel DGC interactors aiming to elucidate the signaling role(s in which the complex is involved. Since the function of the DGC in the brain and nervous system has not been fully defined, we have here continued to analyze the DGC modifiers' function in the developing Drosophila brain and eye. Results Given that disruption of Dys and Dg leads to improper photoreceptor axon projections into the lamina and eye neuron elongation defects during development, we have determined the function of previously screened components and their genetic interaction with the DGC in this tissue. Our study first found that mutations in chif, CG34400, Nrk, Lis1, capt and Cam cause improper axon path-finding and loss of SP2353, Grh, Nrk, capt, CG34400, vimar, Lis1 and Cam cause shortened rhabdomere lengths. We determined that Nrk, mbl, capt and Cam genetically interact with Dys and/or Dg in these processes. It is notable that most of the neuronal DGC interacting components encountered are involved in regulation of actin dynamics. Conclusions Our data indicate possible DGC involvement in the process of cytoskeletal remodeling in neurons. The identification of new components that interact with the DGC not only helps to dissect the mechanism of axon guidance and eye neuron differentiation but also provides a great opportunity for understanding the signaling mechanisms by which the cell surface receptor Dg communicates via Dys with the actin cytoskeleton.

  4. The proton pump inhibitor lansoprazole improves the skeletal phenotype in dystrophin deficient mdx mice.

    Directory of Open Access Journals (Sweden)

    Arpana Sali

    Full Text Available In Duchenne muscular dystrophy (DMD, loss of the membrane stabilizing protein dystrophin results in myofiber damage. Microinjury to dystrophic myofibers also causes secondary imbalances in sarcolemmic ion permeability and resting membrane potential, which modifies excitation-contraction coupling and increases proinflammatory/apoptotic signaling cascades. Although glucocorticoids remain the standard of care for the treatment of DMD, there is a need to investigate the efficacy of other pharmacological agents targeting the involvement of imbalances in ion flux on dystrophic pathology.We designed a preclinical trial to investigate the effects of lansoprazole (LANZO administration, a proton pump inhibitor, on the dystrophic muscle phenotype in dystrophin deficient (mdx mice. Eight to ten week-old female mice were assigned to one of four treatment groups (n = 12 per group: (1 vehicle control; (2 5 mg/kg/day LANZO; (3 5 mg/kg/day prednisolone; and (4 combined treatment of 5 mg/kg/day prednisolone (PRED and 5 mg/kg/day LANZO. Treatment was administered orally 5 d/wk for 3 months. At the end of the study, behavioral (Digiscan and functional outcomes (grip strength and Rotarod were assessed prior to sacrifice. After sacrifice, body, tissue and organ masses, muscle histology, in vitro muscle force, and creatine kinase levels were measured. Mice in the combined treatment groups displayed significant reductions in the number of degenerating muscle fibers and number of inflammatory foci per muscle field relative to vehicle control. Additionally, mice in the combined treatment group displayed less of a decline in normalized forelimb and hindlimb grip strength and declines in in vitro EDL force after repeated eccentric contractions.Together our findings suggest that combined treatment of LANZO and prednisolone attenuates some components of dystrophic pathology in mdx mice. Our findings warrant future investigation of the clinical efficacy of LANZO and

  5. Use of capillary Western immunoassay (Wes) for quantification of dystrophin levels in skeletal muscle of healthy controls and individuals with Becker and Duchenne muscular dystrophy.

    Science.gov (United States)

    Beekman, Chantal; Janson, Anneke A; Baghat, Aabed; van Deutekom, Judith C; Datson, Nicole A

    2018-01-01

    Duchenne muscular dystrophy (DMD) is a neuromuscular disease characterized by progressive weakness of the skeletal and cardiac muscles. This X-linked disorder is caused by open reading frame disrupting mutations in the DMD gene, resulting in strong reduction or complete absence of dystrophin protein. In order to use dystrophin as a supportive or even surrogate biomarker in clinical studies on investigational drugs aiming at correcting the primary cause of the disease, the ability to reliably quantify dystrophin expression in muscle biopsies of DMD patients pre- and post-treatment is essential. Here we demonstrate the application of the ProteinSimple capillary immunoassay (Wes) method, a gel- and blot-free method requiring less sample, antibody and time to run than conventional Western blot assay. We optimized dystrophin quantification by Wes using 2 different antibodies and found it to be highly sensitive, reproducible and quantitative over a large dynamic range. Using a healthy control muscle sample as a reference and α-actinin as a protein loading/muscle content control, a panel of skeletal muscle samples consisting of 31 healthy controls, 25 Becker Muscle dystrophy (BMD) and 17 DMD samples was subjected to Wes analysis. In healthy controls dystrophin levels varied 3 to 5-fold between the highest and lowest muscle samples, with the reference sample representing the average of all 31 samples. In BMD muscle samples dystrophin levels ranged from 10% to 90%, with an average of 33% of the healthy muscle average, while for the DMD samples the average dystrophin level was 1.3%, ranging from 0.7% to 7% of the healthy muscle average. In conclusion, Wes is a suitable, efficient and reliable method for quantification of dystrophin expression as a biomarker in DMD clinical drug development.

  6. MicroRNA-486–dependent modulation of DOCK3/PTEN/AKT signaling pathways improves muscular dystrophy–associated symptoms

    Science.gov (United States)

    Alexander, Matthew S.; Casar, Juan Carlos; Motohashi, Norio; Vieira, Natássia M.; Eisenberg, Iris; Marshall, Jamie L.; Gasperini, Molly J.; Lek, Angela; Myers, Jennifer A.; Estrella, Elicia A.; Kang, Peter B.; Shapiro, Frederic; Rahimov, Fedik; Kawahara, Genri; Widrick, Jeffrey J.; Kunkel, Louis M.

    2014-01-01

    Duchenne muscular dystrophy (DMD) is caused by mutations in the gene encoding dystrophin, which results in dysfunctional signaling pathways within muscle. Previously, we identified microRNA-486 (miR-486) as a muscle-enriched microRNA that is markedly reduced in the muscles of dystrophin-deficient mice (Dmdmdx-5Cv mice) and in DMD patient muscles. Here, we determined that muscle-specific transgenic overexpression of miR-486 in muscle of Dmdmdx-5Cv mice results in reduced serum creatine kinase levels, improved sarcolemmal integrity, fewer centralized myonuclei, increased myofiber size, and improved muscle physiology and performance. Additionally, we identified dedicator of cytokinesis 3 (DOCK3) as a miR-486 target in skeletal muscle and determined that DOCK3 expression is induced in dystrophic muscles. DOCK3 overexpression in human myotubes modulated PTEN/AKT signaling, which regulates muscle hypertrophy and growth, and induced apoptosis. Furthermore, several components of the PTEN/AKT pathway were markedly modulated by miR-486 in dystrophin-deficient muscle. Skeletal muscle–specific miR-486 overexpression in Dmdmdx-5Cv animals decreased levels of DOCK3, reduced PTEN expression, and subsequently increased levels of phosphorylated AKT, which resulted in an overall beneficial effect. Together, these studies demonstrate that stable overexpression of miR-486 ameliorates the disease progression of dystrophin-deficient skeletal muscle. PMID:24789910

  7. Possible influences on the expression of X chromosome-linked dystrophin abnormalities by heterozygosity for autosomal recessive Fukuyama congenital muscular dystrophy

    Energy Technology Data Exchange (ETDEWEB)

    Beggs, A.H.; Neumann, P.E.; Anderson, M.S.; Kunkel, L.M. (Harvard Medical School, Boston, MA (United States)); Arahata, Kiichi; Arikawa, Eri; Nonaka, Ikuya (National Inst. of Neuroscience, Tokyo (Japan))

    1992-01-15

    Abnormalities of dystrophin, a cytoskeletal protein of muscle and nerve, are generally considered specific for Duchenne and Becker muscular dystrophy. However, several patients have recently been identified with dystrophin deficiency who, before dystrophin testing, were considered to have Fukuyama congenital muscular dystrophy (FCMD) on the basis of clinical findings. Epidemiologic data suggest that only 1/3,500 males with autosomal recessive FCMD should have abnormal dystrophin. To explain the observation of 3/23 FCMD males with abnormal dystrophin, the authors propose that dystrophin and the FCMD gene product interact and that the earlier onset and greater severity of these patients' phenotype (relative to Duchenne muscular dystrophy) are due to their being heterozygous for the FCMD mutation in addition to being hemizygous for Duchenne muscular dystrophy, a genotype that is predicted to occur in 1/175,000 Japanese males. This model may help explain the genetic basis for some of the clinical and pathological variability seen among patients with FCMD, and it has potential implications for understanding the inheritance of other autosomal recessive disorders in general. For example, sex ratios for rare autosomal recessive disorders caused by mutations in proteins that interact with X chromosome-linked gene products may display predictable deviation from 1:1.

  8. Deletion of exon 26 of the dystrophin gene is associated with a mild Becker muscular dystrophy phenotype

    DEFF Research Database (Denmark)

    Witting, Nanna; Duno, Morten; Vissing, John

    2011-01-01

    With the possible introduction of exon skipping therapy in Duchenne muscular dystrophy, it has become increasingly important to know the role of each exon of the dystrophin gene to protein expression, and thus the phenotype. In this report, we present two related men with an unusually mild BMD...... calf hypertrophy was noted. Creatine kinase was normal or raised maximally to 500 U/l. The muscle biopsy was myopathic with increased fiber size variation and many internal nuclei, but no dystrophy. No comorbidity was found. In both cases, western blot showed a reduced dystrophin band. Genetic...... skipping therapy for Duchenne muscular dystrophy. This report also shows that BMD may present with a normal CK....

  9. [Clinical features of patients with Becker muscular dystrophy and deletions of the rod domain of dystrophin gene].

    Science.gov (United States)

    Wang, Yanyun; Zhu, Yuling; Yang, Juan; Li, Yaqin; Sun, Jiangwen; Zhan, Yixin; Zhang, Cheng

    2018-02-10

    OBJECTIVE To explore the clinical features of patients carrying deletions of the rod domain of the dystrophin gene. METHODS Clinical data of 12 Chinese patients with Becker muscular dystrophy (BMD) and such deletions was reviewed. RESULTS Most patients complained of muscle weakness of lower limbs. Two patients had muscle cramps, one had increased creatine kinase (CK) level, and one had dilated cardiomyopathy. CONCLUSION Compared with DMD, the clinical features of BMD are much more variable, particularly for those carrying deletions of the rod domain of the dystrophin gene. Muscular weakness may not be the sole complaint of BMD. The diagnosis of BMD cannot be excluded by moderately elevated CK. For male patients with dilated cardiomyopathy, the possibility of BMD should be considered.

  10. Tissue distribution of the dystrophin-related gene product and expression in the mdx and dy mouse

    Energy Technology Data Exchange (ETDEWEB)

    Love, D.R.; Marsden, R.F.; Bloomfield, J.F.; Davies, K.E. (John Radcliffe Hospital, Oxford (England)); Morris, G.E.; Ellis, J.M. (North East Wales Inst., Deeside, Wales (England)); Fairbrother, U.; Edwards, Y.H. (Univ. College London (England)); Slater, C.P. (Newcastle General Hospital, Newcastle-upon-Tyne (England)); Parry, D.J. (Univ. of Ottawa, Ontario (Canada))

    1991-04-15

    The authors have previously reported a dystrophin-related locus (DMDL for Duchenne muscular dystrophy-like) on human chromosome 6 that maps close to the dy mutation on mouse chromosome 10. Here they show that this gene is expressed in a wide range of tissues at varying levels. The transcript is particularly abundant in several human fetal tissues, including heart, placenta, and intestine. Studies with antisera raised against a DMDL fusion protein identify a 400,000 M{sub r} protein in all mouse tissues tested, including those of mdx and dy mice. Unlike the dystrophin gene, the DMDL gene transcript is not differentially spliced at the 3{prime} end in either fetal muscle or brain.

  11. Voluntary wheel running in dystrophin-deficient (mdx) mice: Relationships between exercise parameters and exacerbation of the dystrophic phenotype

    OpenAIRE

    Smythe, Gayle M; White, Jason D

    2012-01-01

    Voluntary wheel running can potentially be used to exacerbate the disease phenotype in dystrophin-deficient mdx mice. While it has been established that voluntary wheel running is highly variable between individuals, the key parameters of wheel running that impact the most on muscle pathology have not been examined in detail. We conducted a 2-week test of voluntary wheel running by mdx mice and the impact of wheel running on disease pathology. There was significant individual variation in the...

  12. Identification of small molecule and genetic modulators of AON-induced dystrophin exon skipping by high-throughput screening.

    Directory of Open Access Journals (Sweden)

    Debra A O'Leary

    Full Text Available One therapeutic approach to Duchenne Muscular Dystrophy (DMD recently entering clinical trials aims to convert DMD phenotypes to that of a milder disease variant, Becker Muscular Dystrophy (BMD, by employing antisense oligonucleotides (AONs targeting splice sites, to induce exon skipping and restore partial dystrophin function. In order to search for small molecule and genetic modulators of AON-dependent and independent exon skipping, we screened approximately 10,000 known small molecule drugs, >17,000 cDNA clones, and >2,000 kinase- targeted siRNAs against a 5.6 kb luciferase minigene construct, encompassing exon 71 to exon 73 of human dystrophin. As a result, we identified several enhancers of exon skipping, acting on both the reporter construct as well as endogenous dystrophin in mdx cells. Multiple mechanisms of action were identified, including histone deacetylase inhibition, tubulin modulation and pre-mRNA processing. Among others, the nucleolar protein NOL8 and staufen RNA binding protein homolog 2 (Stau2 were found to induce endogenous exon skipping in mdx cells in an AON-dependent fashion. An unexpected but recurrent theme observed in our screening efforts was the apparent link between the inhibition of cell cycle progression and the induction of exon skipping.

  13. Optimization of Peptide Nucleic Acid Antisense Oligonucleotides for Local and Systemic Dystrophin Splice Correction in the mdx Mouse

    Science.gov (United States)

    Yin, HaiFang; Betts, Corinne; Saleh, Amer F; Ivanova, Gabriela D; Lee, Hyunil; Seow, Yiqi; Kim, Dalsoo; Gait, Michael J; Wood, Matthew JA

    2010-01-01

    Antisense oligonucleotides (AOs) have the capacity to alter the processing of pre-mRNA transcripts in order to correct the function of aberrant disease-related genes. Duchenne muscular dystrophy (DMD) is a fatal X-linked muscle degenerative disease that arises from mutations in the DMD gene leading to an absence of dystrophin protein. AOs have been shown to restore the expression of functional dystrophin via splice correction by intramuscular and systemic delivery in animal models of DMD and in DMD patients via intramuscular administration. Major challenges in developing this splice correction therapy are to optimize AO chemistry and to develop more effective systemic AO delivery. Peptide nucleic acid (PNA) AOs are an alternative AO chemistry with favorable in vivo biochemical properties and splice correcting abilities. Here, we show long-term splice correction of the DMD gene in mdx mice following intramuscular PNA delivery and effective splice correction in aged mdx mice. Further, we report detailed optimization of systemic PNA delivery dose regimens and PNA AO lengths to yield splice correction, with 25-mer PNA AOs providing the greatest splice correcting efficacy, restoring dystrophin protein in multiple peripheral muscle groups. PNA AOs therefore provide an attractive candidate AO chemistry for DMD exon skipping therapy. PMID:20068555

  14. Dystrophin-deficient dogs with reduced myostatin have unequal muscle growth and greater joint contractures.

    Science.gov (United States)

    Kornegay, Joe N; Bogan, Daniel J; Bogan, Janet R; Dow, Jennifer L; Wang, Jiahui; Fan, Zheng; Liu, Naili; Warsing, Leigh C; Grange, Robert W; Ahn, Mihye; Balog-Alvarez, Cynthia J; Cotten, Steven W; Willis, Monte S; Brinkmeyer-Langford, Candice; Zhu, Hongtu; Palandra, Joe; Morris, Carl A; Styner, Martin A; Wagner, Kathryn R

    2016-01-01

    Myostatin (Mstn) is a negative regulator of muscle growth whose inhibition promotes muscle growth and regeneration. Dystrophin-deficient mdx mice in which myostatin is knocked out or inhibited postnatally have a less severe phenotype with greater total mass and strength and less fibrosis and fatty replacement of muscles than mdx mice with wild-type myostatin expression. Dogs with golden retriever muscular dystrophy (GRMD) have previously been noted to have increased muscle mass and reduced fibrosis after systemic postnatal myostatin inhibition. Based partly on these results, myostatin inhibitors are in development for use in human muscular dystrophies. However, persisting concerns regarding the effects of long-term and profound myostatin inhibition will not be easily or imminently answered in clinical trials. To address these concerns, we developed a canine (GRippet) model by crossbreeding dystrophin-deficient GRMD dogs with Mstn-heterozygous (Mstn (+/-)) whippets. A total of four GRippets (dystrophic and Mstn (+/-)), three GRMD (dystrophic and Mstn wild-type) dogs, and three non-dystrophic controls from two litters were evaluated. Myostatin messenger ribonucleic acid (mRNA) and protein levels were downregulated in both GRMD and GRippet dogs. GRippets had more severe postural changes and larger (more restricted) maximal joint flexion angles, apparently due to further exaggeration of disproportionate effects on muscle size. Flexors such as the cranial sartorius were more hypertrophied on magnetic resonance imaging (MRI) in the GRippets, while extensors, including the quadriceps femoris, underwent greater atrophy. Myostatin protein levels negatively correlated with relative cranial sartorius muscle cross-sectional area on MRI, supporting a role in disproportionate muscle size. Activin receptor type IIB (ActRIIB) expression was higher in dystrophic versus control dogs, consistent with physiologic feedback between myostatin and ActRIIB. However, there was no

  15. Aberrant location of inhibitory synaptic marker proteins in the hippocampus of dystrophin-deficient mice: implications for cognitive impairment in duchenne muscular dystrophy.

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    Elżbieta Krasowska

    Full Text Available Duchenne muscular dystrophy (DMD is a neuromuscular disease that arises from mutations in the dystrophin-encoding gene. Apart from muscle pathology, cognitive impairment, primarily of developmental origin, is also a significant component of the disorder. Convergent lines of evidence point to an important role for dystrophin in regulating the molecular machinery of central synapses. The clustering of neurotransmitter receptors at inhibitory synapses, thus impacting on synaptic transmission, is of particular significance. However, less is known about the role of dystrophin in influencing the precise expression patterns of proteins located within the pre- and postsynaptic elements of inhibitory synapses. To this end, we exploited molecular markers of inhibitory synapses, interneurons and dystrophin-deficient mouse models to explore the role of dystrophin in determining the stereotypical patterning of inhibitory connectivity within the cellular networks of the hippocampus CA1 region. In tissue from wild-type (WT mice, immunoreactivity of neuroligin2 (NL2, an adhesion molecule expressed exclusively in postsynaptic elements of inhibitory synapses, and the vesicular GABA transporter (VGAT, a marker of GABAergic presynaptic elements, were predictably enriched in strata pyramidale and lacunosum moleculare. In acute contrast, NL2 and VGAT immunoreactivity was relatively evenly distributed across all CA1 layers in dystrophin-deficient mice. Similar changes were evident with the cannabinoid receptor 1, vesicular glutamate transporter 3, parvalbumin, somatostatin and the GABAA receptor alpha1 subunit. The data show that in the absence of dystrophin, there is a rearrangement of the molecular machinery, which underlies the precise spatio-temporal pattern of GABAergic synaptic transmission within the CA1 sub-field of the hippocampus.

  16. Serum cholinesterases are differentially regulated in normal and dystrophin-deficient mutant mice

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    Andrea R. Durrant

    2012-06-01

    Full Text Available The cholinesterases, acetylcholinesterase and butyrylcholinesterase (pseudocholinesterase, are abundant in the nervous system and in other tissues. The role of acetylcholinesterase in terminating transmitter action in the peripheral and central nervous system is well understood. However, both knowledge of the function(s of the cholinesterases in serum, and of their metabolic and endocrine regulation under normal and pathological conditions, is limited. This study investigates acetylcholinesterase and butyrylcholinesterase in sera of dystrophin-deficient mdx mutant mice, an animal model for the human Duchenne muscular dystrophy and in control healthy mice. The data show systematic and differential variations in the concentrations of both enzymes in the sera, and specific changes dictated by alteration of hormonal balance in both healthy and dystrophic mice. While acetylcholinesterase in mdx-sera is elevated, butyrylcholinesterase is markedly diminished, resulting in an overall cholinesterase decrease compared to sera of healthy controls. The androgen testosterone (T is a negative modulator of butyrylcholinesterase, but not of acetylcholinesterase, in male mouse sera. T-removal elevated both butyrylcholinesterase activity and the butyrylcholinesterase/acetylcholinesterase ratio in mdx male sera to values resembling those in healthy control male mice. Mechanisms of regulation of the circulating cholinesterases and their impairment in the dystrophic mice are suggested, and clinical implications for diagnosis and treatment are considered.

  17. Metabolic remodeling agents show beneficial effects in the dystrophin-deficient mdx mouse model

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    Jahnke Vanessa E

    2012-08-01

    Full Text Available Abstract Background Duchenne muscular dystrophy is a genetic disease involving a severe muscle wasting that is characterized by cycles of muscle degeneration/regeneration and culminates in early death in affected boys. Mitochondria are presumed to be involved in the regulation of myoblast proliferation/differentiation; enhancing mitochondrial activity with exercise mimetics (AMPK and PPAR-delta agonists increases muscle function and inhibits muscle wasting in healthy mice. We therefore asked whether metabolic remodeling agents that increase mitochondrial activity would improve muscle function in mdx mice. Methods Twelve-week-old mdx mice were treated with two different metabolic remodeling agents (GW501516 and AICAR, separately or in combination, for 4 weeks. Extensive systematic behavioral, functional, histological, biochemical, and molecular tests were conducted to assess the drug(s' effects. Results We found a gain in body and muscle weight in all treated mice. Histologic examination showed a decrease in muscle inflammation and in the number of fibers with central nuclei and an increase in fibers with peripheral nuclei, with significantly fewer activated satellite cells and regenerating fibers. Together with an inhibition of FoXO1 signaling, these results indicated that the treatments reduced ongoing muscle damage. Conclusions The three treatments produced significant improvements in disease phenotype, including an increase in overall behavioral activity and significant gains in forelimb and hind limb strength. Our findings suggest that triggering mitochondrial activity with exercise mimetics improves muscle function in dystrophin-deficient mdx mice.

  18. Recombinase-mediated reprogramming and dystrophin gene addition in mdx mouse induced pluripotent stem cells.

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    Chunli Zhao

    Full Text Available A cell therapy strategy utilizing genetically-corrected induced pluripotent stem cells (iPSC may be an attractive approach for genetic disorders such as muscular dystrophies. Methods for genetic engineering of iPSC that emphasize precision and minimize random integration would be beneficial. We demonstrate here an approach in the mdx mouse model of Duchenne muscular dystrophy that focuses on the use of site-specific recombinases to achieve genetic engineering. We employed non-viral, plasmid-mediated methods to reprogram mdx fibroblasts, using phiC31 integrase to insert a single copy of the reprogramming genes at a safe location in the genome. We next used Bxb1 integrase to add the therapeutic full-length dystrophin cDNA to the iPSC in a site-specific manner. Unwanted DNA sequences, including the reprogramming genes, were then precisely deleted with Cre resolvase. Pluripotency of the iPSC was analyzed before and after gene addition, and ability of the genetically corrected iPSC to differentiate into myogenic precursors was evaluated by morphology, immunohistochemistry, qRT-PCR, FACS analysis, and intramuscular engraftment. These data demonstrate a non-viral, reprogramming-plus-gene addition genetic engineering strategy utilizing site-specific recombinases that can be applied easily to mouse cells. This work introduces a significant level of precision in the genetic engineering of iPSC that can be built upon in future studies.

  19. RT-PCR analysis of dystrophin mRNA in DND/BMD patients

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    Ciafaloni, E.; Silva, H.A.R. de; Roses, A.D. [Duke Univ. Medical Center, Durham, NC (United States)

    1994-09-01

    Duchenne and Becker muscular dystrophies (DMD, BMD) are X-linked recessive disorders caused by mutations in the dystrophin (dys) gene. The majority of these mutations are intragenic deletions of duplications routinely detected by Southern biots and multiplex PCR. The remainder are very likely, smaller mutations, mostly point-mutations. Detection of these mutations is very difficult due to the size and complexity of the dys gene. We applied RT-PCR to analyse the entire dys mRNA of three DMD patients with no detectable genomic defect. In two unrelated patients, a duplication of the 62 bp exon 2 was identified. This causes a frameshift sufficient to explain the DMD phenotype. In the third patient, who had congenital DMD and severe mental retardation, a complex pattern of aberrant splicing at the 3-prime exons 67-79 was observed. Sural nerve biopsy in this patient showed the complete absence of Dp116. PCR-SSCP studies are presently in progress to identify the mutations responsible for the aberrant splicing patterns.

  20. Somatodendritic and excitatory postsynaptic distribution of neuron-type dystrophin isoform, Dp40, in hippocampal neurons.

    Science.gov (United States)

    Fujimoto, Takahiro; Itoh, Kyoko; Yaoi, Takeshi; Fushiki, Shinji

    2014-09-12

    The Duchenne muscular dystrophy (DMD) gene produces multiple dystrophin (Dp) products due to the presence of several promoters. We previously reported the existence of a novel short isoform of Dp, Dp40, in adult mouse brain. However, the exact biochemical expression profile and cytological distribution of the Dp40 protein remain unknown. In this study, we generated a polyclonal antibody against the NH2-terminal region of the Dp40 and identified the expression profile of Dp40 in the mouse brain. Through an analysis using embryonic and postnatal mouse cerebrums, we found that Dp40 emerged from the early neonatal stages until adulthood, whereas Dp71, an another Dp short isoform, was highly detected in both prenatal and postnatal cerebrums. Intriguingly, relative expressions of Dp40 and Dp71 were prominent in cultured dissociated neurons and non-neuronal cells derived from mouse hippocampus, respectively. Furthermore, the immunocytological distribution of Dp40 was analyzed in dissociated cultured neurons, revealing that Dp40 is detected in the soma and its dendrites, but not in the axon. It is worthy to note that Dp40 is localized along the subplasmalemmal region of the dendritic shafts, as well as at excitatory postsynaptic sites. Thus, Dp40 was identified as a neuron-type Dp possibly involving dendritic and synaptic functions. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Sequence characterisation of deletion breakpoints in the dystrophin gene by PCR

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    Abbs, S.; Sandhu, S.; Bobrow, M. [Guy`s Hospital, London (United Kingdom)

    1994-09-01

    Partial deletions of the dystrophin gene account for 65% of cases of Duchenne muscular dystrophy. A high proportion of these structural changes are generated by new mutational events, and lie predominantly within two `hotspot` regions, yet the underlying reasons for this are not known. We are characterizing and sequencing the regions surrounding deletion breakpoints in order to: (i) investigate the mechanisms of deletion mutation, and (ii) enable the design of PCR assays to specifically amplify mutant and normal sequences, allowing us to search for the presence of somatic mosaicism in appropriate family members. Using this approach we have been able to demonstrate the presence of somatic mosaicism in a maternal grandfather of a DMD-affected male, deleted for exons 49-50. Three deletions, namely of exons 48-49, 49-50, and 50, have been characterized using a PCR approach that avoids any cloning procedures. Breakpoints were initially localized to within regions of a few kilobases using Southern blot restriction analyses with exon-specific probes and PCR amplification of exonic and intronic loci. Sequencing was performed directly on PCR products: (i) mutant sequences were obtained from long-range or inverse-PCR across the deletion junction fragments, and (ii) normal sequences were obtained from the products of standard PCR, vectorette PCR, or inverse-PCR performed on YACs. Further characterization of intronic sequences will allow us to amplify and sequence across other deletion breakpoints and increase our knowledge of the mechanisms of mutation in the dystophin gene.

  2. Haplotypes in the dystrophin DNA segment point to a mosaic origin of modern human diversity.

    Science.gov (United States)

    Zietkiewicz, Ewa; Yotova, Vania; Gehl, Dominik; Wambach, Tina; Arrieta, Isabel; Batzer, Mark; Cole, David E C; Hechtman, Peter; Kaplan, Feige; Modiano, David; Moisan, Jean-Paul; Michalski, Roman; Labuda, Damian

    2003-11-01

    Although Africa has played a central role in human evolutionary history, certain studies have suggested that not all contemporary human genetic diversity is of recent African origin. We investigated 35 simple polymorphic sites and one T(n) microsatellite in an 8-kb segment of the dystrophin gene. We found 86 haplotypes in 1,343 chromosomes from around the world. Although a classical out-of-Africa topology was observed in trees based on the variant frequencies, the tree of haplotype sequences reveals three lineages accounting for present-day diversity. The proportion of new recombinants and the diversity of the T(n) microsatellite were used to estimate the age of haplotype lineages and the time of colonization events. The lineage that underwent the great expansion originated in Africa prior to the Upper Paleolithic (27,000-56,000 years ago). A second group, of structurally distinct haplotypes that occupy a central position on the tree, has never left Africa. The third lineage is represented by the haplotype that lies closest to the root, is virtually absent in Africa, and appears older than the recent out-of-Africa expansion. We propose that this lineage could have left Africa before the expansion (as early as 160,000 years ago) and admixed, outside of Africa, with the expanding lineage. Contemporary human diversity, although dominated by the recently expanded African lineage, thus represents a mosaic of different contributions.

  3. Simultaneous Pathoproteomic Evaluation of the Dystrophin-Glycoprotein Complex and Secondary Changes in the mdx-4cv Mouse Model of Duchenne Muscular Dystrophy

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    Sandra Murphy

    2015-06-01

    Full Text Available In skeletal muscle, the dystrophin-glycoprotein complex forms a membrane-associated assembly of relatively low abundance, making its detailed proteomic characterization in normal versus dystrophic tissues technically challenging. To overcome this analytical problem, we have enriched the muscle membrane fraction by a minimal differential centrifugation step followed by the comprehensive label-free mass spectrometric analysis of microsomal membrane preparations. This organelle proteomic approach successfully identified dystrophin and its binding partners in normal versus dystrophic hind limb muscles. The introduction of a simple pre-fractionation step enabled the simultaneous proteomic comparison of the reduction in the dystrophin-glycoprotein complex and secondary changes in the mdx-4cv mouse model of dystrophinopathy in a single analytical run. The proteomic screening of the microsomal fraction from dystrophic hind limb muscle identified the full-length dystrophin isoform Dp427 as the most drastically reduced protein in dystrophinopathy, demonstrating the remarkable analytical power of comparative muscle proteomics. Secondary pathoproteomic expression patterns were established for 281 proteins, including dystrophin-associated proteins and components involved in metabolism, signalling, contraction, ion-regulation, protein folding, the extracellular matrix and the cytoskeleton. Key findings were verified by immunoblotting. Increased levels of the sarcolemmal Na+/K+-ATPase in dystrophic leg muscles were also confirmed by immunofluorescence microscopy. Thus, the reduction of sample complexity in organelle-focused proteomics can be advantageous for the profiling of supramolecular protein complexes in highly intricate systems, such as skeletal muscle tissue.

  4. Consecutive analysis of mutation spectrum in the dystrophin gene of 507 Korean boys with Duchenne/Becker muscular dystrophy in a single center.

    Science.gov (United States)

    Cho, Anna; Seong, Moon-Woo; Lim, Byung Chan; Lee, Hwa Jeen; Byeon, Jung Hye; Kim, Seung Soo; Kim, Soo Yeon; Choi, Sun Ah; Wong, Ai-Lynn; Lee, Jeongho; Kim, Jon Soo; Ryu, Hye Won; Lee, Jin Sook; Kim, Hunmin; Hwang, Hee; Choi, Ji Eun; Kim, Ki Joong; Hwang, Young Seung; Hong, Ki Ho; Park, Seungman; Cho, Sung Im; Lee, Seung Jun; Park, Hyunwoong; Seo, Soo Hyun; Park, Sung Sup; Chae, Jong Hee

    2017-05-01

    Duchenne and Becker muscular dystrophies (DMD and BMD) are allelic X-linked recessive muscle diseases caused by mutations in the large and complex dystrophin gene. We analyzed the dystrophin gene in 507 Korean DMD/BMD patients by multiple ligation-dependent probe amplification and direct sequencing. Overall, 117 different deletions, 48 duplications, and 90 pathogenic sequence variations, including 30 novel variations, were identified. Deletions and duplications accounted for 65.4% and 13.3% of Korean dystrophinopathy, respectively, suggesting that the incidence of large rearrangements in dystrophin is similar among different ethnic groups. We also detected sequence variations in >100 probands. The small variations were dispersed across the whole gene, and 12.3% were nonsense mutations. Precise genetic characterization in patients with DMD/BMD is timely and important for implementing nationwide registration systems and future molecular therapeutic trials in Korea and globally. Muscle Nerve 55: 727-734, 2017. © 2016 Wiley Periodicals, Inc.

  5. Micro Engineering

    DEFF Research Database (Denmark)

    Alting, Leo; Kimura, F.; Hansen, Hans Nørgaard

    2003-01-01

    The paper addresses the questions of how micro products are designed and how they are manufactured. Definitions of micro products and micro engineering are discussed and the presentation is aimed at describing typical issues, possibilities and tools regarding design of micro products. The implica......The paper addresses the questions of how micro products are designed and how they are manufactured. Definitions of micro products and micro engineering are discussed and the presentation is aimed at describing typical issues, possibilities and tools regarding design of micro products...

  6. MLPA based detection of mutations in the dystrophin gene of 180 Polish families with Duchenne/Becker muscular dystrophy.

    Science.gov (United States)

    Zimowski, Janusz G; Massalska, Diana; Holding, Mariola; Jadczak, Sylwia; Fidziańska, Elżbieta; Lusakowska, Anna; Kostera-Pruszczyk, Anna; Kamińska, Anna; Zaremba, Jacek

    2014-01-01

    Duchenne/Becker muscular dystrophy (DMD/BMD) is a recessive, X-linked disorder caused by a mutation in the dystrophin gene. Deletions account for approximately 60-65% of mutations, duplications for 5-10%. The remaining cases are mainly point mutations. According to Monaco theory clinical form of the disease depends on maintaining or disrupting the reading frame. The purpose of the study was to determine frequency and location of deletions and duplications in the dystrophin gene, to determine the compliance between maintaining/disrupting the reading frame and clinical form of the disease and to check the effectiveness of MLPA (multiplex ligation-dependent probe amplification) in the detection of these mutations in hemizygous patients and heterozygous female carriers. The material is composed of combined results of molecular diagnosis carried out in years 2009-2012 in 180 unrelated patients referred with the diagnosis of DMD/BMD tested by use of MLPA. We identified 110 deletions, 22 duplication (in one patient two different duplications were detected) and 2 point mutations. Deletions involved mainly exons 45-54 and 3-21, whereas most duplications involved exons 3-18. The compliance with Monaco theory was 95% for deletions and 76% for duplications. Most of mutations in the dystrophin gene were localized in the hot spots - different for deletions and duplications. MLPA enabled their quick identification, exact localization and determination whether or not they maintained or disrupted the reading frame. MLPA was also effective in detection of deletions and duplications in female carriers. Copyright © 2014 Polish Neurological Society. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  7. Dystrophin is required for the normal function of the cardio-protective K(ATP channel in cardiomyocytes.

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    Laura Graciotti

    Full Text Available Duchenne and Becker muscular dystrophy patients often develop a cardiomyopathy for which the pathogenesis is still unknown. We have employed the murine animal model of Duchenne muscular dystrophy (mdx, which develops a cardiomyopathy that includes some characteristics of the human disease, to study the molecular basis of this pathology. Here we show that the mdx mouse heart has defects consistent with alteration in compounds that regulate energy homeostasis including a marked decrease in creatine-phosphate (PC. In addition, the mdx heart is more susceptible to anoxia than controls. Since the cardio-protective ATP sensitive potassium channel (K(ATP complex and PC have been shown to interact we investigated whether deficits in PC levels correlate with other molecular events including K(ATP ion channel complex presence, its functionality and interaction with dystrophin. We found that this channel complex is present in the dystrophic cardiac cell membrane but its ability to sense a drop in the intracellular ATP concentration and consequently open is compromised by the absence of dystrophin. We further demonstrate that the creatine kinase muscle isoform (CKm is displaced from the plasma membrane of the mdx cardiac cells. Considering that CKm is a determinant of K(ATP channel complex function we hypothesize that dystrophin acts as a scaffolding protein organizing the K(ATP channel complex and the enzymes necessary for its correct functioning. Therefore, the lack of proper functioning of the cardio-protective K(ATP system in the mdx cardiomyocytes may be part of the mechanism contributing to development of cardiac disease in dystrophic patients.

  8. Diseased muscles that lack dystrophin or laminin-α2 have altered compositions and proliferation of mononuclear cell populations

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    Miller Jeffrey

    2005-04-01

    Full Text Available Abstract Background Multiple types of mononucleate cells reside among the multinucleate myofibers in skeletal muscles and these mononucleate cells function in muscle maintenance and repair. How neuromuscular disease might affect different types of muscle mononucleate cells had not been determined. In this study, therefore, we examined how two neuromuscular diseases, dystrophin-deficiency and laminin-α2-deficiency, altered the proliferation and composition of different subsets of muscle-derived mononucleate cells. Methods We used fluorescence-activated cell sorting combined with bromodeoxyuridine labeling to examine proliferation rates and compositions of mononuclear cells in diseased and healthy mouse skeletal muscle. We prepared mononucleate cells from muscles of mdx (dystrophin-deficient or Lama2-/- (laminin-α2-deficient mice and compared them to cells from healthy control muscles. We enumerated subsets of resident muscle cells based on Sca-1 and CD45 expression patterns and determined the proliferation of each cell subset in vivo by BrdU incorporation. Results We found that the proliferation and composition of the mononucleate cells in dystrophin-deficient and laminin-α2-deficient diseased muscles are different than in healthy muscle. The mdx and Lama2-/- muscles showed similar significant increases in CD45+ cells compared to healthy muscle. Changes in proliferation, however, differed between the two diseases with proliferation increased in mdx and decreased in Lama2-/- muscles compared to healthy muscles. In particular, the most abundant Sca-1-/CD45- subset, which contains muscle precursor cells, had increased proliferation in mdx muscle but decreased proliferation in Lama2-/- muscles. Conclusion The similar increases in CD45+ cells, but opposite changes in proliferation of muscle precursor cells, may underlie aspects of the distinct pathologies in the two diseases.

  9. Astrogliosis and impaired aquaporin-4 and dystrophin systems in idiopathic normal pressure hydrocephalus.

    Science.gov (United States)

    Eide, P K; Hansson, H-A

    2017-06-19

    Idiopathic normal pressure hydrocephalus (iNPH) is one subtype of dementia that may improve following drainage of cerebrospinal fluid (CSF). This prospective observational study explored whether expression of the water channel aquaporin-4 (AQP4) and the anchoring molecule dystrophin 71 (Dp71) are altered at astrocytic perivascular endfeet and in adjacent neuropil of iNPH patient. Observations were related to measurements of pulsatile and static intracranial pressure (ICP). The study included iNPH patients undergoing overnight monitoring of the pulsatile/static ICP in whom a biopsy was taken from the frontal cerebral cortex during placement of the ICP sensor. Reference (Ref) biopsies were sampled from 13 patients who underwent brain surgery for epilepsy, tumours or cerebral aneurysms. The brain tissue specimens were examined by light microscopy, immunohistochemistry, densitometry and morphometry. iNPH patients responding to surgery (n = 44) had elevated pulsatile ICP, indicative of impaired intracranial compliance. As compared to the Ref patients, the cortical biopsies of iNPH patients revealed prominent astrogliosis and reduced expression of AQP4 and Dp71 immunoreactivities in the astrocytic perivascular endfeet and in parts of the adjacent neuropil. There was a significant correlation between degree of astrogliosis and reduction of AQP4 and Dp71 at astrocytic perivascular endfeet. Idiopathic normal pressure hydrocephalus patients responding to CSF diversion present with abnormal pulsatile ICP, indicative of impaired intracranial compliance. A main histopathological finding was astrogliosis and reduction of AQP4 and of Dp71 in astrocytic perivascular endfeet. We propose that the altered AQP4 and Dp71 complex contributes to the subischaemia prevalent in the brain tissue of iNPH. © 2017 British Neuropathological Society.

  10. Sensitivity and Frequencies of Dystrophin Gene Mutations in Thai DMD/BMD Patients As Detected by Multiplex PCR

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    Thanyachai Sura

    2008-01-01

    Full Text Available Background: Duchenne muscular dystrophy (DMD, a lethal X-linked disease affecting 1 in 3500 male births, and its more benign variant, Becker muscular dystrophy (BMD, are caused by mutations in the dystrophin gene. Because of its large size, analysing the whole gene is impractical. Methods have been developed to detect the commonest mutations i.e. the deletions of the exons. Although these tests are highly specific, their sensitivity is inherently limited by the prevalence of deletions, which differs among different populations.

  11. GRAF1 deficiency blunts sarcolemmal injury repair and exacerbates cardiac and skeletal muscle pathology in dystrophin-deficient mice.

    Science.gov (United States)

    Lenhart, Kaitlin C; O'Neill, Thomas J; Cheng, Zhaokang; Dee, Rachel; Demonbreun, Alexis R; Li, Jianbin; Xiao, Xiao; McNally, Elizabeth M; Mack, Christopher P; Taylor, Joan M

    2015-01-01

    The plasma membranes of striated muscle cells are particularly susceptible to rupture as they endure significant mechanical stress and strain during muscle contraction, and studies have shown that defects in membrane repair can contribute to the progression of muscular dystrophy. The synaptotagmin-related protein, dysferlin, has been implicated in mediating rapid membrane repair through its ability to direct intracellular vesicles to sites of membrane injury. However, further work is required to identify the precise molecular mechanisms that govern dysferlin targeting and membrane repair. We previously showed that the bin-amphiphysin-Rvs (BAR)-pleckstrin homology (PH) domain containing Rho-GAP GTPase regulator associated with focal adhesion kinase-1 (GRAF1) was dynamically recruited to the tips of fusing myoblasts wherein it promoted membrane merging by facilitating ferlin-dependent capturing of intracellular vesicles. Because acute membrane repair responses involve similar vesicle trafficking complexes/events and because our prior studies in GRAF1-deficient tadpoles revealed a putative role for GRAF1 in maintaining muscle membrane integrity, we postulated that GRAF1 might also play an important role in facilitating dysferlin-dependent plasma membrane repair. We used an in vitro laser-injury model to test whether GRAF1 was necessary for efficient muscle membrane repair. We also generated dystrophin/GRAF1 doubledeficient mice by breeding mdx mice with GRAF1 hypomorphic mice. Evans blue dye uptake and extensive morphometric analyses were used to assess sarcolemmal integrity and related pathologies in cardiac and skeletal muscles isolated from these mice. Herein, we show that GRAF1 is dynamically recruited to damaged skeletal and cardiac muscle plasma membranes and that GRAF1-depleted muscle cells have reduced membrane healing abilities. Moreover, we show that dystrophin depletion exacerbated muscle damage in GRAF1-deficient mice and that mice with dystrophin/GRAF1

  12. Exonization of an Intronic LINE-1 Element Causing Becker Muscular Dystrophy as a Novel Mutational Mechanism in Dystrophin Gene.

    Science.gov (United States)

    Gonçalves, Ana; Oliveira, Jorge; Coelho, Teresa; Taipa, Ricardo; Melo-Pires, Manuel; Sousa, Mário; Santos, Rosário

    2017-10-03

    A broad mutational spectrum in the dystrophin ( DMD ) gene, from large deletions/duplications to point mutations, causes Duchenne/Becker muscular dystrophy (D/BMD). Comprehensive genotyping is particularly relevant considering the mutation-centered therapies for dystrophinopathies. We report the genetic characterization of a patient with disease onset at age 13 years, elevated creatine kinase levels and reduced dystrophin labeling, where multiplex-ligation probe amplification (MLPA) and genomic sequencing failed to detect pathogenic variants. Bioinformatic, transcriptomic (real time PCR, RT-PCR), and genomic approaches (Southern blot, long-range PCR, and single molecule real-time sequencing) were used to characterize the mutation. An aberrant transcript was identified, containing a 103-nucleotide insertion between exons 51 and 52, with no similarity with the DMD gene. This corresponded to the partial exonization of a long interspersed nuclear element (LINE-1), disrupting the open reading frame. Further characterization identified a complete LINE-1 (~6 kb with typical hallmarks) deeply inserted in intron 51. Haplotyping and segregation analysis demonstrated that the mutation had a de novo origin. Besides underscoring the importance of mRNA studies in genetically unsolved cases, this is the first report of a disease-causing fully intronic LINE-1 element in DMD , adding to the diversity of mutational events that give rise to D/BMD.

  13. Exonization of an Intronic LINE-1 Element Causing Becker Muscular Dystrophy as a Novel Mutational Mechanism in Dystrophin Gene

    Science.gov (United States)

    Gonçalves, Ana; Coelho, Teresa; Melo-Pires, Manuel; Sousa, Mário

    2017-01-01

    A broad mutational spectrum in the dystrophin (DMD) gene, from large deletions/duplications to point mutations, causes Duchenne/Becker muscular dystrophy (D/BMD). Comprehensive genotyping is particularly relevant considering the mutation-centered therapies for dystrophinopathies. We report the genetic characterization of a patient with disease onset at age 13 years, elevated creatine kinase levels and reduced dystrophin labeling, where multiplex-ligation probe amplification (MLPA) and genomic sequencing failed to detect pathogenic variants. Bioinformatic, transcriptomic (real time PCR, RT-PCR), and genomic approaches (Southern blot, long-range PCR, and single molecule real-time sequencing) were used to characterize the mutation. An aberrant transcript was identified, containing a 103-nucleotide insertion between exons 51 and 52, with no similarity with the DMD gene. This corresponded to the partial exonization of a long interspersed nuclear element (LINE-1), disrupting the open reading frame. Further characterization identified a complete LINE-1 (~6 kb with typical hallmarks) deeply inserted in intron 51. Haplotyping and segregation analysis demonstrated that the mutation had a de novo origin. Besides underscoring the importance of mRNA studies in genetically unsolved cases, this is the first report of a disease-causing fully intronic LINE-1 element in DMD, adding to the diversity of mutational events that give rise to D/BMD. PMID:28972564

  14. 100-fold but not 50-fold dystrophin overexpression aggravates electrocardiographic defects in the mdx model of Duchenne muscular dystrophy

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    Yongping Yue

    2016-01-01

    Full Text Available Dystrophin gene replacement holds the promise of treating Duchenne muscular dystrophy. Supraphysiological expression is a concern for all gene therapy studies. In the case of Duchenne muscular dystrophy, Chamberlain and colleagues found that 50-fold overexpression did not cause deleterious side effect in skeletal muscle. To determine whether excessive dystrophin expression in the heart is safe, we studied two lines of transgenic mdx mice that selectively expressed a therapeutic minidystrophin gene in the heart at 50-fold and 100-fold of the normal levels. In the line with 50-fold overexpression, minidystrophin showed sarcolemmal localization and electrocardiogram abnormalities were corrected. However, in the line with 100-fold overexpression, we not only detected sarcolemmal minidystrophin expression but also observed accumulation of minidystrophin vesicles in the sarcoplasm. Excessive minidystrophin expression did not correct tachycardia, a characteristic feature of Duchenne muscular dystrophy. Importantly, several electrocardiogram parameters (QT interval, QRS duration and the cardiomyopathy index became worse than that of mdx mice. Our data suggests that the mouse heart can tolerate 50-fold minidystrophin overexpression, but 100-fold overexpression leads to cardiac toxicity.

  15. Eosinophilia of dystrophin-deficient muscle is promoted by perforin-mediated cytotoxicity by T cell effectors

    Science.gov (United States)

    Cai, B.; Spencer, M. J.; Nakamura, G.; Tseng-Ong, L.; Tidball, J. G.

    2000-01-01

    Previous investigations have shown that cytotoxic T lymphocytes (CTLs) contribute to muscle pathology in the dystrophin-null mutant mouse (mdx) model of Duchenne muscular dystrophy through perforin-dependent and perforin-independent mechanisms. We have assessed whether the CTL-mediated pathology includes the promotion of eosinophilia in dystrophic muscle, and thereby provides a secondary mechanism through which CTLs contribute to muscular dystrophy. Quantitative immunohistochemistry confirmed that eosinophilia is a component of the mdx dystrophy. In addition, electron microscopic observations show that eosinophils traverse the basement membrane of mdx muscle fibers and display sites of close apposition of eosinophil and muscle membranes. The close membrane apposition is characterized by impingement of eosinophilic rods of major basic protein into the muscle cell membrane. Transfer of mdx splenocytes and mdx muscle extracts to irradiated C57 mice by intraperitoneal injection resulted in muscle eosinophilia in the recipient mice. Double-mutant mice lacking dystrophin and perforin showed less eosinophilia than was displayed by mdx mice that expressed perforin. Finally, administration of prednisolone, which has been shown previously to reduce the concentration of CTLs in dystrophic muscle, produced a significant reduction in eosinophilia. These findings indicate that eosinophilia is a component of the mdx pathology that is promoted by perforin-dependent cytotoxicity of effector T cells. However, some eosinophilia of mdx muscle is independent of perforin-mediated processes.

  16. Short (16-mer locked nucleic acid splice-switching oligonucleotides restore dystrophin production in Duchenne Muscular Dystrophy myotubes.

    Directory of Open Access Journals (Sweden)

    Vanessa Borges Pires

    Full Text Available Splice-switching antisense oligonucleotides (SSOs offer great potential for RNA-targeting therapies, and two SSO drugs have been recently approved for treating Duchenne Muscular Dystrophy (DMD and Spinal Muscular Atrophy (SMA. Despite promising results, new developments are still needed for more efficient chemistries and delivery systems. Locked nucleic acid (LNA is a chemically modified nucleic acid that presents several attractive properties, such as high melting temperature when bound to RNA, potent biological activity, high stability and low toxicity in vivo. Here, we designed a series of LNA-based SSOs complementary to two sequences of the human dystrophin exon 51 that are most evolutionary conserved and evaluated their ability to induce exon skipping upon transfection into myoblasts derived from a DMD patient. We show that 16-mers with 60% of LNA modification efficiently induce exon skipping and restore synthesis of a truncated dystrophin isoform that localizes to the plasma membrane of patient-derived myotubes differentiated in culture. In sum, this study underscores the value of short LNA-modified SSOs for therapeutic applications.

  17. Nanopatterned muscle cell patches for enhanced myogenesis and dystrophin expression in a mouse model of muscular dystrophy.

    Science.gov (United States)

    Yang, Hee Seok; Ieronimakis, Nicholas; Tsui, Jonathan H; Kim, Hong Nam; Suh, Kahp-Yang; Reyes, Morayma; Kim, Deok-Ho

    2014-02-01

    Skeletal muscle is a highly organized tissue in which the extracellular matrix (ECM) is composed of highly-aligned cables of collagen with nanoscale feature sizes, and provides structural and functional support to muscle fibers. As such, the transplantation of disorganized tissues or the direct injection of cells into muscles for regenerative therapy often results in suboptimal functional improvement due to a failure to integrate with native tissue properly. Here, we present a simple method in which biodegradable, biomimetic substrates with precisely controlled nanotopography were fabricated using solvent-assisted capillary force lithography (CFL) and were able to induce the proper development and differentiation of primary mononucleated cells to form mature muscle patches. Cells cultured on these nanopatterned substrates were highly-aligned and elongated, and formed more mature myotubes as evidenced by up-regulated expression of the myogenic regulatory factors Myf5, MyoD and myogenin (MyoG). When transplanted into mdx mice models for Duchenne muscular dystrophy (DMD), the proposed muscle patches led to the formation of a significantly greater number of dystrophin-positive muscle fibers, indicating that dystrophin replacement and myogenesis is achievable in vivo with this approach. These results demonstrate the feasibility of utilizing biomimetic substrates not only as platforms for studying the influences of the ECM on skeletal muscle function and maturation, but also to create transplantable muscle cell patches for the treatment of chronic and acute muscle diseases or injuries. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Immobilization of Dystrophin and Laminin α2-Chain Deficient Zebrafish Larvae In Vivo Prevents the Development of Muscular Dystrophy.

    Directory of Open Access Journals (Sweden)

    Mei Li

    Full Text Available Muscular dystrophies are often caused by genetic alterations in the dystrophin-dystroglycan complex or its extracellular ligands. These structures are associated with the cell membrane and provide mechanical links between the cytoskeleton and the matrix. Mechanical stress is considered a pathological mechanism and muscle immobilization has been shown to be beneficial in some mouse models of muscular dystrophy. The zebrafish enables novel and less complex models to examine the effects of extended immobilization or muscle relaxation in vivo in different dystrophy models. We have examined effects of immobilization in larvae from two zebrafish strains with muscular dystrophy, the Sapje dystrophin-deficient and the Candyfloss laminin α2-chain-deficient strains. Larvae (4 days post fertilization, dpf of both mutants have significantly lower active force in vitro, alterations in the muscle structure with gaps between muscle fibers and altered birefringence patterns compared to their normal siblings. Complete immobilization (18 hrs to 4 dpf was achieved using a small molecular inhibitor of actin-myosin interaction (BTS, 50 μM. This treatment resulted in a significantly weaker active contraction at 4 dpf in both mutated larvae and normal siblings, most likely reflecting a general effect of immobilization on myofibrillogenesis. The immobilization also significantly reduced the structural damage in the mutated strains, showing that muscle activity is an important pathological mechanism. Following one-day washout of BTS, muscle tension partly recovered in the Candyfloss siblings and caused structural damage in these mutants, indicating activity-induced muscle recovery and damage, respectively.

  19. Gentamicin treatment in exercised mdx mice: Identification of dystrophin-sensitive pathways and evaluation of efficacy in work-loaded dystrophic muscle.

    Science.gov (United States)

    De Luca, Annamaria; Nico, Beatrice; Rolland, Jean-François; Cozzoli, Anna; Burdi, Rosa; Mangieri, Domenica; Giannuzzi, Viviana; Liantonio, Antonella; Cippone, Valentina; De Bellis, Michela; Nicchia, Grazia Paola; Camerino, Giulia Maria; Frigeri, Antonio; Svelto, Maria; Camerino, Diana Conte

    2008-11-01

    Aminoglycosides force read through of premature stop codon mutations and introduce new mutation-specific gene-corrective strategies in Duchenne muscular dystrophy. A chronic treatment with gentamicin (32 mg/kg/daily i.p., 8-12 weeks) was performed in exercised mdx mice with the dual aim to clarify the dependence on dystrophin of the functional, biochemical and histological alterations present in dystrophic muscle and to verify the long term efficiency of small molecule gene-corrective strategies in work-loaded dystrophic muscle. The treatment counteracted the exercise-induced impairment of in vivo forelimb strength after 6-8 weeks. We observed an increase in dystrophin expression level in all the fibers, although lower than that observed in normal fibers, and found a concomitant recovery of aquaporin-4 at sarcolemma. A significant reduction in centronucleated fibers, in the area of necrosis and in the percentage of nuclear factor-kB-positive nuclei was observed in gastrocnemious muscle of treated animals. Plasma creatine kinase was reduced by 70%. Ex vivo, gentamicin restored membrane ionic conductance in mdx diaphragm and limb muscle fibers. No effects were observed on the altered calcium homeostasis and sarcolemmal calcium permeability, detected by electrophysiological and microspectrofluorimetric approaches. Thus, the maintenance of a partial level of dystrophin is sufficient to reinforce sarcolemmal stability, reducing leakiness, inflammation and fiber damage, while correction of altered calcium homeostasis needs greater expression of dystrophin or direct interventions on the channels involved.

  20. Protein truncation test: analysis of two novel point mutations at the carboxy-terminus of the human dystrophin gene associated with mental retardation.

    Science.gov (United States)

    Tuffery, S; Lenk, U; Roberts, R G; Coubes, C; Demaille, J; Claustres, M

    1995-01-01

    Approximately one-third of the mutations responsible for Duchenne muscular dytrophy (DMD) do not involve gross rearrangements of the dystrophin gene. Methods for intensive mutation screening have recently been applied to this immense gene, which resulted in the identification of a number of point mutations in DMD patients, mostly translation-terminating mutations. A number of data raised the possibility that the C-terminal region of dystrophin might be involved in some cases of mental retardation associated with DMD. Using single-strand conformation analysis of products amplified by polymerase chain reaction (PCR-SSCA) to screen the terminal domains of the dystrophin gene (exons 60-79) of 20 unrelated patients with DMD or BMD, we detected two novel point mutations in two mentally retarded DMD patients: a 1-bp deletion in exon 70 (10334delC) and a 5' splice donor site alteration in intron 69 (10294 + 1G-->T). Both mutations should result in a premature translation termination of dystrophin. The possible effects on the reading frame were analyzed by the study of reverse transcripts amplified from peripheral blood lymphocytes mRNA and by the protein truncation test.

  1. Concurrent Label-Free Mass Spectrometric Analysis of Dystrophin Isoform Dp427 and the Myofibrosis Marker Collagen in Crude Extracts from mdx-4cv Skeletal Muscles

    Science.gov (United States)

    Murphy, Sandra; Zweyer, Margit; Mundegar, Rustam R.; Henry, Michael; Meleady, Paula; Swandulla, Dieter; Ohlendieck, Kay

    2015-01-01

    The full-length dystrophin protein isoform of 427 kDa (Dp427), the absence of which represents the principal abnormality in X-linked muscular dystrophy, is difficult to identify and characterize by routine proteomic screening approaches of crude tissue extracts. This is probably related to its large molecular size, its close association with the sarcolemmal membrane, and its existence within a heterogeneous glycoprotein complex. Here, we used a careful extraction procedure to isolate the total protein repertoire from normal versus dystrophic mdx-4cv skeletal muscles, in conjunction with label-free mass spectrometry, and successfully identified Dp427 by proteomic means. In contrast to a considerable number of previous comparative studies of the total skeletal muscle proteome, using whole tissue proteomics we show here for the first time that the reduced expression of this membrane cytoskeletal protein is the most significant alteration in dystrophinopathy. This agrees with the pathobiochemical concept that the almost complete absence of dystrophin is the main defect in Duchenne muscular dystrophy and that the mdx-4cv mouse model of dystrophinopathy exhibits only very few revertant fibers. Significant increases in collagens and associated fibrotic marker proteins, such as fibronectin, biglycan, asporin, decorin, prolargin, mimecan, and lumican were identified in dystrophin-deficient muscles. The up-regulation of collagen in mdx-4cv muscles was confirmed by immunofluorescence microscopy and immunoblotting. Thus, this is the first mass spectrometric study of crude tissue extracts that puts the proteomic identification of dystrophin in its proper pathophysiological context. PMID:28248273

  2. Somatic mosaicism of a point mutation in the dystrophin gene in a patient presenting with an asymmetrical muscle weakness and contractures

    NARCIS (Netherlands)

    Helderman-van den Enden, A. T. J. M.; Ginjaar, H. B.; Kneppers, A. L. J.; Bakker, E.; Breuning, M. H.; de Visser, M.

    2003-01-01

    We describe a patient with somatic mosaicism of a point mutation in the dystrophin gene causing benign muscular dystrophy with an unusual asymmetrical distribution of muscle weakness and contractures. To our knowledge this is the first patient with asymmetrical weakness and contractures in an

  3. Fetal microchimeric cells in a fetus-treats-its-mother paradigm do not contribute to dystrophin production in serially parous mdx females.

    Science.gov (United States)

    Seppanen, Elke Jane; Hodgson, Samantha Susan; Khosrotehrani, Kiarash; Bou-Gharios, George; Fisk, Nicholas M

    2012-10-10

    Throughout every pregnancy, genetically distinct fetal microchimeric stem/progenitor cells (FMCs) engraft in the mother, persist long after delivery, and may home to damaged maternal tissues. Phenotypically normal fetal lymphoid progenitors have been described to develop in immunodeficient mothers in a fetus-treats-its-mother paradigm. Since stem cells contribute to muscle repair, we assessed this paradigm in the mdx mouse model of Duchenne muscular dystrophy. mdx females were bred serially to either ROSAeGFP males or mdx males to obtain postpartum microchimeras that received either wild-type FMCs or dystrophin-deficient FMCs through serial gestations. To enhance regeneration, notexin was injected into the tibialis anterior of postpartum mice. FMCs were detected by qPCR at a higher frequency in injected compared to noninjected side muscle (P=0.02). However, the number of dystrophin-positive fibers was similar in mothers delivering wild-type compared to mdx pups. In addition, there was no correlation between FMC detection and percentage dystrophin, and no GFP+ve FMCs were identified that expressed dystrophin. In 10/11 animals, GFP+ve FMCs were detected by immunohistochemistry, of which 60% expressed CD45 with 96% outside the basal lamina defining myofiber contours. Finally we confirmed lack of FMC contribution to statellite cells in postpartum mdx females mated with Myf5-LacZ males. We conclude that the FMC contribution to regenerating muscles is insufficient to have a functional impact.

  4. More deletions in the 5{prime} region than in the central region of the dystrophin gene were identified among Filipino Duchenne and Becker muscular dystrophy patients

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-11-06

    This report describes mutations in the dystrophin gene and the frequency of these mutations in Filipino pedigrees with Duchenne and Becker muscular dystrophy (DMD/BMD). The findings suggest the presence of genetic variability among DMD/BMD patients in different populations. 13 refs., 1 tab.

  5. Identification of a novel first exon in the human dystrophin gene and of a new promoter located more than 500 kb upstream of the nearest known promoter

    Energy Technology Data Exchange (ETDEWEB)

    Yanagawa, H.; Nishio, H.; Takeshima, Y. [Kobe Univ. School of Medicine (Japan)] [and others

    1994-09-01

    The dystrophin gene, which is muted in patients with Duchenne and Becker muscular dystrophies, is the largest known human gene. Five alternative promoters have been characterized until now. Here we show that a novel dystrophin isoform with a different first exon can be produced through transcription initiation at a previously-unidentified alternative promoter. The case study presented is that of patient with Duchenne muscular dystrophy who had a deletion extending from 5{prime} end of the dystrophin gene to exon 2, including all promoters previously mapped in the 5{prime} part of the gene. Transcripts from lymphoblastoid cells were found to contain sequences corresponding to exon 3, indicating the presence of new promoter upstream of this exon. The nucleotide sequence of amplified cDNA corresponding to the 5{prime} end of the new transcript indicated that the 5{prime} end of exon 3 was extended by 9 codons, only the last (most 3{prime}) of which codes for methionine. The genomic nucleotide sequence upstream from the new exon, as determined using inverse polymerase chain reaction, revealed the presence of sequences similar to a TATA box, an octamer motif and an MEF-2 element. The identified promoter/exon did not map to intron 2, as might have been expected, but to a position more than 500 kb upstream of the most 5{prime} of the previously-identified promoters, thereby adding 500 kb to the dystrophin gene. The sequence of part of the new promoter region is very similar to that of certain medium reiteration frequency repetitive sequences. These findings may help us understand the molecular evolution of the dystrophin gene.

  6. Long-term rescue of dystrophin expression and improvement in muscle pathology and function in dystrophic mdx mice by peptide-conjugated morpholino.

    Science.gov (United States)

    Wu, Bo; Lu, Peijuan; Cloer, Caryn; Shaban, Mona; Grewal, Snimar; Milazi, Stephanie; Shah, Sapana N; Moulton, Hong M; Lu, Qi Long

    2012-08-01

    Exon skipping is capable of correcting frameshift and nonsense mutations in Duchenne muscular dystrophy. Phase 2 clinical trials in the United Kingdom and the Netherlands have reported induction of dystrophin expression in muscle of Duchenne muscular dystrophy patients by systemic administration of both phosphorodiamidate morpholino oligomers (PMO) and 2'-O-methyl phosphorothioate. Peptide-conjugated phosphorodiamidate morpholino offers significantly higher efficiency than phosphorodiamidate morpholino, with the ability to induce near-normal levels of dystrophin, and restores function in both skeletal and cardiac muscle. We examined 1-year systemic efficacy of peptide-conjugated phosphorodiamidate morpholino targeting exon 23 in dystrophic mdx mice. The LD(50) of peptide-conjugated phosphorodiamidate morpholino was determined to be approximately 85 mg/kg. The half-life of dystrophin expression was approximately 2 months in skeletal muscle, but shorter in cardiac muscle. Biweekly injection of 6 mg/kg peptide-conjugated phosphorodiamidate morpholino produced >20% dystrophin expression in all skeletal muscles and ≤5% in cardiac muscle, with improvement in muscle function and pathology and reduction in levels of serum creatine kinase. Monthly injections of 30 mg/kg peptide-conjugated phosphorodiamidate morpholino restored dystrophin to >50% normal levels in skeletal muscle, and 15% in cardiac muscle. This was associated with greatly reduced serum creatine kinase levels, near-normal histology, and functional improvement of skeletal muscle. Our results demonstrate for the first time that regular 1-year administration of peptide-conjugated phosphorodiamidate morpholino can be safely applied to achieve significant therapeutic effects in an animal model. Copyright © 2012 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

  7. Micro Vision

    OpenAIRE

    Ohba, Kohtaro; Ohara, Kenichi

    2007-01-01

    In the field of the micro vision, there are few researches compared with macro environment. However, applying to the study result for macro computer vision technique, you can measure and observe the micro environment. Moreover, based on the effects of micro environment, it is possible to discovery the new theories and new techniques.

  8. Evaluation of point mutations in dystrophin gene in Iranian Duchenne and Becker muscular dystrophy patients: introducing three novel variants.

    Science.gov (United States)

    Haghshenas, Maryam; Akbari, Mohammad Taghi; Karizi, Shohreh Zare; Deilamani, Faravareh Khordadpoor; Nafissi, Shahriar; Salehi, Zivar

    2016-06-01

    Duchenne and Becker muscular dystrophies (DMD and BMD) are X-linked neuromuscular diseases characterized by progressive muscular weakness and degeneration of skeletal muscles. Approximately two-thirds of the patients have large deletions or duplications in the dystrophin gene and the remaining one-third have point mutations. This study was performed to evaluate point mutations in Iranian DMD/BMD male patients. A total of 29 DNA samples from patients who did not show any large deletion/duplication mutations following multiplex polymerase chain reaction (PCR) and multiplex ligation-dependent probe amplification (MLPA) screening were sequenced for detection of point mutations in exons 50-79. Also exon 44 was sequenced in one sample in which a false positive deletion was detected by MLPA method. Cycle sequencing revealed four nonsense, one frameshift and two splice site mutations as well as two missense variants.

  9. Deletion of exon 26 of the dystrophin gene is associated with a mild Becker muscular dystrophy phenotype

    DEFF Research Database (Denmark)

    Witting, Nanna; Duno, Morten; Vissing, John

    2011-01-01

    With the possible introduction of exon skipping therapy in Duchenne muscular dystrophy, it has become increasingly important to know the role of each exon of the dystrophin gene to protein expression, and thus the phenotype. In this report, we present two related men with an unusually mild BMD...... skipping therapy for Duchenne muscular dystrophy. This report also shows that BMD may present with a normal CK....... associated with an exon 26 deletion. The proband, a 23-year-old man, had slightly delayed motor milestones, walking 1 1/2 years old. He had no complaints of muscle weakness, but had muscle pain. Clinical examination revealed no muscle wasting or loss of power, but his CK was 1500-7000 U/l. Muscle biopsy...

  10. Dystrophin Hot-Spot Mutants Leading to Becker Muscular Dystrophy Insert More Deeply into Membrane Models than the Native Protein.

    Science.gov (United States)

    Ameziane-Le Hir, Sarah; Paboeuf, Gilles; Tascon, Christophe; Hubert, Jean-François; Le Rumeur, Elisabeth; Vié, Véronique; Raguénès-Nicol, Céline

    2016-07-26

    Dystrophin (DYS) is a membrane skeleton protein whose mutations lead to lethal Duchenne muscular dystrophy or to the milder Becker muscular dystrophy (BMD). One third of BMD "in-frame" exon deletions are located in the region that codes for spectrin-like repeats R16 to R21. We focused on four prevalent mutated proteins deleted in this area (called RΔ45-47, RΔ45-48, RΔ45-49, and RΔ45-51 according to the deleted exon numbers), analyzing protein/membrane interactions. Two of the mutants, RΔ45-48 and RΔ45-51, led to mild pathologies and displayed a similar triple coiled-coil structure as the full-length DYS R16-21, whereas the two others, RΔ45-47 and RΔ45-49, induced more severe pathologies and showed "fractional" structures unrelated to the normal one. To explore lipid packing, small unilamellar liposomes (SUVs) and planar monolayers were used at various initial surface pressures. The dissociation constants determined by microscale thermophoresis (MST) were much higher for the full-length DYS R161-21 than for the mutants; thus the wild type protein has weaker SUV binding. Comparing surface pressures after protein adsorption and analysis of atomic force microscopy images of mixed protein/lipid monolayers revealed that the mutants insert more into the lipid monolayer than the wild type does. In fact, in both models every deletion mutant showed more interactions with membranes than the full-length protein did. This means that mutations in the R16-21 part of dystrophin disturb the protein's molecular behavior as it relates to membranes, regardless of whether the accompanying pathology is mild or severe.

  11. Defects in mitochondrial ATP synthesis in dystrophin-deficient mdx skeletal muscles may be caused by complex I insufficiency.

    Directory of Open Access Journals (Sweden)

    Emma Rybalka

    Full Text Available Duchenne Muscular Dystrophy is a chronic, progressive and ultimately fatal skeletal muscle wasting disease characterised by sarcolemmal fragility and intracellular Ca2+ dysregulation secondary to the absence of dystrophin. Mounting literature also suggests that the dysfunction of key energy systems within the muscle may contribute to pathological muscle wasting by reducing ATP availability to Ca2+ regulation and fibre regeneration. No study to date has biochemically quantified and contrasted mitochondrial ATP production capacity by dystrophic mitochondria isolated from their pathophysiological environment such to determine whether mitochondria are indeed capable of meeting this heightened cellular ATP demand, or examined the effects of an increasing extramitochondrial Ca2+ environment. Using isolated mitochondria from the diaphragm and tibialis anterior of 12 week-old dystrophin-deficient mdx and healthy control mice (C57BL10/ScSn we have demonstrated severely depressed Complex I-mediated mitochondrial ATP production rate in mdx mitochondria that occurs irrespective of the macronutrient-derivative substrate combination fed into the Kreb's cycle, and, which is partially, but significantly, ameliorated by inhibition of Complex I with rotenone and stimulation of Complex II-mediated ATP-production with succinate. There was no difference in the MAPR response of mdx mitochondria to increasing extramitochondrial Ca2+ load in comparison to controls, and 400 nM extramitochondrial Ca2+ was generally shown to be inhibitory to MAPR in both groups. Our data suggests that DMD pathology is exacerbated by a Complex I deficiency, which may contribute in part to the severe reductions in ATP production previously observed in dystrophic skeletal muscle.

  12. Sparing of the dystrophin-deficient cranial sartorius muscle is associated with classical and novel hypertrophy pathways in GRMD dogs.

    Science.gov (United States)

    Nghiem, Peter P; Hoffman, Eric P; Mittal, Priya; Brown, Kristy J; Schatzberg, Scott J; Ghimbovschi, Svetlana; Wang, Zuyi; Kornegay, Joe N

    2013-11-01

    Both Duchenne and golden retriever muscular dystrophy (GRMD) are caused by dystrophin deficiency. The Duchenne muscular dystrophy sartorius muscle and orthologous GRMD cranial sartorius (CS) are relatively spared/hypertrophied. We completed hierarchical clustering studies to define molecular mechanisms contributing to this differential involvement and their role in the GRMD phenotype. GRMD dogs with larger CS muscles had more severe deficits, suggesting that selective hypertrophy could be detrimental. Serial biopsies from the hypertrophied CS and other atrophied muscles were studied in a subset of these dogs. Myostatin showed an age-dependent decrease and an inverse correlation with the degree of GRMD CS hypertrophy. Regulators of myostatin at the protein (AKT1) and miRNA (miR-539 and miR-208b targeting myostatin mRNA) levels were altered in GRMD CS, consistent with down-regulation of myostatin signaling, CS hypertrophy, and functional rescue of this muscle. mRNA and proteomic profiling was used to identify additional candidate genes associated with CS hypertrophy. The top-ranked network included α-dystroglycan and like-acetylglucosaminyltransferase. Proteomics demonstrated increases in myotrophin and spectrin that could promote hypertrophy and cytoskeletal stability, respectively. Our results suggest that multiple pathways, including decreased myostatin and up-regulated miRNAs, α-dystroglycan/like-acetylglucosaminyltransferase, spectrin, and myotrophin, contribute to hypertrophy and functional sparing of the CS. These data also underscore the muscle-specific responses to dystrophin deficiency and the potential deleterious effects of differential muscle involvement. Copyright © 2013 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

  13. Clinical and genetic characterisation of dystrophin-deficient muscular dystrophy in a family of Miniature Poodle dogs.

    Directory of Open Access Journals (Sweden)

    Lluís Sánchez

    Full Text Available Four full-sibling intact male Miniature Poodles were evaluated at 4-19 months of age. One was clinically normal and three were affected. All affected dogs were reluctant to exercise and had generalised muscle atrophy, a stiff gait and a markedly elevated serum creatine kinase activity. Two affected dogs also showed poor development, learning difficulties and episodes of abnormal behaviour. In these two dogs, investigations into forebrain structural and metabolic diseases were unremarkable; electromyography demonstrated fibrillation potentials and complex repetitive discharges in the infraspinatus, supraspinatus and epaxial muscles. Histopathological, immunohistochemical and immunoblotting analyses of muscle biopsies were consistent with dystrophin-deficient muscular dystrophy. DNA samples were obtained from all four full-sibling male Poodles, a healthy female littermate and the dam, which was clinically normal. Whole genome sequencing of one affected dog revealed a >5 Mb deletion on the X chromosome, encompassing the entire DMD gene. The exact deletion breakpoints could not be experimentally ascertained, but we confirmed that this region was deleted in all affected males, but not in the unaffected dogs. Quantitative polymerase chain reaction confirmed all three affected males were hemizygous for the mutant X chromosome, while the wildtype chromosome was observed in the unaffected male littermate. The female littermate and the dam were both heterozygous for the mutant chromosome. Forty-four Miniature Poodles from the general population were screened for the mutation and were homozygous for the wildtype chromosome. The finding represents a naturally-occurring mutation causing dystrophin-deficient muscular dystrophy in the dog.

  14. Motor physical therapy affects muscle collagen type I and decreases gait speed in dystrophin-deficient dogs.

    Directory of Open Access Journals (Sweden)

    Thaís P Gaiad

    Full Text Available Golden Retriever Muscular Dystrophy (GRMD is a dystrophin-deficient canine model genetically homologous to Duchenne Muscular Dystrophy (DMD in humans. Muscular fibrosis secondary to cycles of degeneration/regeneration of dystrophic muscle tissue and muscular weakness leads to biomechanical adaptation that impairs the quality of gait. Physical therapy (PT is one of the supportive therapies available for DMD, however, motor PT approaches have controversial recommendations and there is no consensus regarding the type and intensity of physical therapy. In this study we investigated the effect of physical therapy on gait biomechanics and muscular collagen deposition types I and III in dystrophin-deficient dogs. Two dystrophic dogs (treated dogs-TD underwent a PT protocol of active walking exercise, 3×/week, 40 minutes/day, 12 weeks. Two dystrophic control dogs (CD maintained their routine of activities of daily living. At t0 (pre and t1 (post-physical therapy, collagen type I and III were assessed by immunohistochemistry and gait biomechanics were analyzed. Angular displacement of shoulder, elbow, carpal, hip, stifle and tarsal joint and vertical (Fy, mediolateral (Fz and craniocaudal (Fx ground reaction forces (GRF were assessed. Wilcoxon test was used to verify the difference of biomechanical variables between t0 and t1, considering p<.05. Type I collagen of endomysium suffered the influence of PT, as well as gait speed that had decreased from t0 to t1 (p<.000. The PT protocol employed accelerates morphological alterations on dystrophic muscle and promotes a slower velocity of gait. Control dogs which maintained their routine of activities of daily living seem to have found a better balance between movement and preservation of motor function.

  15. Motor Physical Therapy Affects Muscle Collagen Type I and Decreases Gait Speed in Dystrophin-Deficient Dogs

    Science.gov (United States)

    Gaiad, Thaís P.; Araujo, Karla P. C.; Serrão, Júlio C.; Miglino, Maria A.; Ambrósio, Carlos Eduardo

    2014-01-01

    Golden Retriever Muscular Dystrophy (GRMD) is a dystrophin-deficient canine model genetically homologous to Duchenne Muscular Dystrophy (DMD) in humans. Muscular fibrosis secondary to cycles of degeneration/regeneration of dystrophic muscle tissue and muscular weakness leads to biomechanical adaptation that impairs the quality of gait. Physical therapy (PT) is one of the supportive therapies available for DMD, however, motor PT approaches have controversial recommendations and there is no consensus regarding the type and intensity of physical therapy. In this study we investigated the effect of physical therapy on gait biomechanics and muscular collagen deposition types I and III in dystrophin-deficient dogs. Two dystrophic dogs (treated dogs-TD) underwent a PT protocol of active walking exercise, 3×/week, 40 minutes/day, 12 weeks. Two dystrophic control dogs (CD) maintained their routine of activities of daily living. At t0 (pre) and t1 (post-physical therapy), collagen type I and III were assessed by immunohistochemistry and gait biomechanics were analyzed. Angular displacement of shoulder, elbow, carpal, hip, stifle and tarsal joint and vertical (Fy), mediolateral (Fz) and craniocaudal (Fx) ground reaction forces (GRF) were assessed. Wilcoxon test was used to verify the difference of biomechanical variables between t0 and t1, considering p<.05. Type I collagen of endomysium suffered the influence of PT, as well as gait speed that had decreased from t0 to t1 (p<.000). The PT protocol employed accelerates morphological alterations on dystrophic muscle and promotes a slower velocity of gait. Control dogs which maintained their routine of activities of daily living seem to have found a better balance between movement and preservation of motor function. PMID:24713872

  16. Evaluation of skeletal and cardiac muscle function after chronic administration of thymosin beta-4 in the dystrophin deficient mouse.

    Directory of Open Access Journals (Sweden)

    Christopher F Spurney

    2010-01-01

    Full Text Available Thymosin beta-4 (Tbeta4 is a ubiquitous protein with many properties relating to cell proliferation and differentiation that promotes wound healing and modulates inflammatory mediators. We studied the effects of chronic administration of Tbeta4 on the skeletal and cardiac muscle of dystrophin deficient mdx mice, the mouse model of Duchenne muscular dystrophy. Female wild type (C57BL10/ScSnJ and mdx mice, 8-10 weeks old, were treated with 150 microg of Tbeta4 twice a week for 6 months. To promote muscle pathology, mice were exercised for 30 minutes twice a week. Skeletal and cardiac muscle function were assessed via grip strength and high frequency echocardiography. Localization of Tbeta4 and amount of fibrosis were quantified using immunohistochemistry and Gomori's tri-chrome staining, respectively. Mdx mice treated with Tbeta4 showed a significant increase in skeletal muscle regenerating fibers compared to untreated mdx mice. Tbeta4 stained exclusively in the regenerating fibers of mdx mice. Although untreated mdx mice had significantly decreased skeletal muscle strength compared to untreated wild type, there were no significant improvements in mdx mice after treatment. Systolic cardiac function, measured as percent shortening fraction, was decreased in untreated mdx mice compared to untreated wild type and there was no significant difference after treatment in mdx mice. Skeletal and cardiac muscle fibrosis were also significantly increased in untreated mdx mice compared to wild type, but there was no significant improvement in treated mdx mice. In exercised dystrophin deficient mice, chronic administration of Tbeta4 increased the number of regenerating fibers in skeletal muscle and could have a potential role in treatment of skeletal muscle disease in Duchenne muscular dystrophy.

  17. Distal mdx muscle groups exhibiting up-regulation of utrophin and rescue of dystrophin-associated glycoproteins exemplify a protected phenotype in muscular dystrophy

    Science.gov (United States)

    Dowling, Paul; Culligan, Kevin; Ohlendieck, Kay

    2002-02-01

    Unique unaffected skeletal muscle fibres, unlike necrotic torso and limb muscles, may pave the way for a more detailed understanding of the molecular pathogenesis of inherited neuromuscular disorders and help to develop new treatment strategies for muscular dystrophies. The sparing of extraocular muscle in Duchenne muscular dystrophy is mostly attributed to the special protective properties of extremely fast-twitching small-diameter fibres, but here we show that distal muscles also represent a particular phenotype that is more resistant to necrosis. Immunoblot analysis of membranes isolated from the well established dystrophic animal model mdx shows that, in contrast to dystrophic limb muscles, the toe musculature exhibits an up-regulation of the autosomal dystrophin homologue utrophin and a concomitant rescue of dystrophin-associated glycoproteins. Thus distal mdx muscle groups provide a cellular system that naturally avoids myofibre degeneration which might be useful in the search for naturally occurring compensatory mechanisms in inherited skeletal muscle diseases.

  18. Correlation of Utrophin Levels with the Dystrophin Protein Complex and Muscle Fibre Regeneration in Duchenne and Becker Muscular Dystrophy Muscle Biopsies.

    Science.gov (United States)

    Janghra, Narinder; Morgan, Jennifer E; Sewry, Caroline A; Wilson, Francis X; Davies, Kay E; Muntoni, Francesco; Tinsley, Jonathon

    2016-01-01

    Duchenne muscular dystrophy is a severe and currently incurable progressive neuromuscular condition, caused by mutations in the DMD gene that result in the inability to produce dystrophin. Lack of dystrophin leads to loss of muscle fibres and a reduction in muscle mass and function. There is evidence from dystrophin-deficient mouse models that increasing levels of utrophin at the muscle fibre sarcolemma by genetic or pharmacological means significantly reduces the muscular dystrophy pathology. In order to determine the efficacy of utrophin modulators in clinical trials, it is necessary to accurately measure utrophin levels and other biomarkers on a fibre by fibre basis within a biopsy section. Our aim was to develop robust and reproducible staining and imaging protocols to quantify sarcolemmal utrophin levels, sarcolemmal dystrophin complex members and numbers of regenerating fibres within a biopsy section. We quantified sarcolemmal utrophin in mature and regenerating fibres and the percentage of regenerating muscle fibres, in muscle biopsies from Duchenne, the milder Becker muscular dystrophy and controls. Fluorescent immunostaining followed by image analysis was performed to quantify utrophin intensity and β-dystrogylcan and ɣ -sarcoglycan intensity at the sarcolemma. Antibodies to fetal and developmental myosins were used to identify regenerating muscle fibres allowing the accurate calculation of percentage regeneration fibres in the biopsy. Our results indicate that muscle biopsies from Becker muscular dystrophy patients have fewer numbers of regenerating fibres and reduced utrophin intensity compared to muscle biopsies from Duchenne muscular dystrophy patients. Of particular interest, we show for the first time that the percentage of regenerating muscle fibres within the muscle biopsy correlate with the clinical severity of Becker and Duchenne muscular dystrophy patients from whom the biopsy was taken. The ongoing development of these tools to quantify

  19. Correlation of Utrophin Levels with the Dystrophin Protein Complex and Muscle Fibre Regeneration in Duchenne and Becker Muscular Dystrophy Muscle Biopsies.

    Directory of Open Access Journals (Sweden)

    Narinder Janghra

    Full Text Available Duchenne muscular dystrophy is a severe and currently incurable progressive neuromuscular condition, caused by mutations in the DMD gene that result in the inability to produce dystrophin. Lack of dystrophin leads to loss of muscle fibres and a reduction in muscle mass and function. There is evidence from dystrophin-deficient mouse models that increasing levels of utrophin at the muscle fibre sarcolemma by genetic or pharmacological means significantly reduces the muscular dystrophy pathology. In order to determine the efficacy of utrophin modulators in clinical trials, it is necessary to accurately measure utrophin levels and other biomarkers on a fibre by fibre basis within a biopsy section. Our aim was to develop robust and reproducible staining and imaging protocols to quantify sarcolemmal utrophin levels, sarcolemmal dystrophin complex members and numbers of regenerating fibres within a biopsy section. We quantified sarcolemmal utrophin in mature and regenerating fibres and the percentage of regenerating muscle fibres, in muscle biopsies from Duchenne, the milder Becker muscular dystrophy and controls. Fluorescent immunostaining followed by image analysis was performed to quantify utrophin intensity and β-dystrogylcan and ɣ -sarcoglycan intensity at the sarcolemma. Antibodies to fetal and developmental myosins were used to identify regenerating muscle fibres allowing the accurate calculation of percentage regeneration fibres in the biopsy. Our results indicate that muscle biopsies from Becker muscular dystrophy patients have fewer numbers of regenerating fibres and reduced utrophin intensity compared to muscle biopsies from Duchenne muscular dystrophy patients. Of particular interest, we show for the first time that the percentage of regenerating muscle fibres within the muscle biopsy correlate with the clinical severity of Becker and Duchenne muscular dystrophy patients from whom the biopsy was taken. The ongoing development of these

  20. A duchenne muscular dystrophy gene hot spot mutation in dystrophin-deficient cavalier king charles spaniels is amenable to exon 51 skipping.

    Directory of Open Access Journals (Sweden)

    Gemma L Walmsley

    2010-01-01

    Full Text Available Duchenne muscular dystrophy (DMD, which afflicts 1 in 3500 boys, is one of the most common genetic disorders of children. This fatal degenerative condition is caused by an absence or deficiency of dystrophin in striated muscle. Most affected patients have inherited or spontaneous deletions in the dystrophin gene that disrupt the reading frame resulting in unstable truncated products. For these patients, restoration of the reading frame via antisense oligonucleotide-mediated exon skipping is a promising therapeutic approach. The major DMD deletion "hot spot" is found between exons 45 and 53, and skipping exon 51 in particular is predicted to ameliorate the dystrophic phenotype in the greatest number of patients. Currently the mdx mouse is the most widely used animal model of DMD, although its mild phenotype limits its suitability in clinical trials. The Golden Retriever muscular dystrophy (GRMD model has a severe phenotype, but due to its large size, is expensive to use. Both these models have mutations in regions of the dystrophin gene distant from the commonly mutated DMD "hot spot".Here we describe the severe phenotype, histopathological findings, and molecular analysis of Cavalier King Charles Spaniels with dystrophin-deficient muscular dystrophy (CKCS-MD. The dogs harbour a missense mutation in the 5' donor splice site of exon 50 that results in deletion of exon 50 in mRNA transcripts and a predicted premature truncation of the translated protein. Antisense oligonucleotide-mediated skipping of exon 51 in cultured myoblasts from an affected dog restored the reading frame and protein expression.Given the small size of the breed, the amiable temperament and the nature of the mutation, we propose that CKCS-MD is a valuable new model for clinical trials of antisense oligonucleotide-induced exon skipping and other therapeutic approaches for DMD.

  1. Effective myotube formation in human adipose tissue-derived stem cells expressing dystrophin and myosin heavy chain by cellular fusion with mouse C2C12 myoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Eom, Young Woo [Cell Therapy and Tissue Engineering Center, Wonju College of Medicine, Yonsei Univ., Wonju (Korea, Republic of); Biomedical Research Institute, Lifeliver Co., Ltd., Suwon (Korea, Republic of); Lee, Jong Eun; Yang, Mal Sook; Jang, In Keun; Kim, Hyo Eun; Lee, Doo Hoon; Kim, Young Jin [Biomedical Research Institute, Lifeliver Co., Ltd., Suwon (Korea, Republic of); Park, Won Jin [Dr. Park' s Aesthetic Clinic, Seoul (Korea, Republic of); Kong, Jee Hyun; Shim, Kwang Yong [Department of Hematology-Oncology, Wonju College of Medicine, Yonsei Univ., Wonju (Korea, Republic of); Lee, Jong In, E-mail: oncochem@yonsei.ac.kr [Department of Hematology-Oncology, Wonju College of Medicine, Yonsei Univ., Wonju (Korea, Republic of); Kim, Hyun Soo, E-mail: khsmd@unitel.co.kr [Department of Hematology-Oncology, Wonju College of Medicine, Yonsei Univ., Wonju (Korea, Republic of)

    2011-04-29

    Highlights: {yields} hASCs were differentiated into skeletal muscle cells by treatment with 5-azacytidine, FGF-2, and the supernatant of cultured hASCs. {yields} Dystrophin and MyHC were expressed in late differentiation step by treatment with the supernatant of cultured hASCs. {yields} hASCs expressing dystrophin and MyHC contributed to myotube formation during co-culture with mouse myoblast C2C12 cells. -- Abstract: Stem cell therapy for muscular dystrophies requires stem cells that are able to participate in the formation of new muscle fibers. However, the differentiation steps that are the most critical for this process are not clear. We investigated the myogenic phases of human adipose tissue-derived stem cells (hASCs) step by step and the capability of myotube formation according to the differentiation phase by cellular fusion with mouse myoblast C2C12 cells. In hASCs treated with 5-azacytidine and fibroblast growth factor-2 (FGF-2) for 1 day, the early differentiation step to express MyoD and myogenin was induced by FGF-2 treatment for 6 days. Dystrophin and myosin heavy chain (MyHC) expression was induced by hASC conditioned medium in the late differentiation step. Myotubes were observed only in hASCs undergoing the late differentiation step by cellular fusion with C2C12 cells. In contrast, hASCs that were normal or in the early stage were not involved in myotube formation. Our results indicate that stem cells expressing dystrophin and MyHC are more suitable for myotube formation by co-culture with myoblasts than normal or early differentiated stem cells expressing MyoD and myogenin.

  2. Effective myotube formation in human adipose tissue-derived stem cells expressing dystrophin and myosin heavy chain by cellular fusion with mouse C2C12 myoblasts

    International Nuclear Information System (INIS)

    Eom, Young Woo; Lee, Jong Eun; Yang, Mal Sook; Jang, In Keun; Kim, Hyo Eun; Lee, Doo Hoon; Kim, Young Jin; Park, Won Jin; Kong, Jee Hyun; Shim, Kwang Yong; Lee, Jong In; Kim, Hyun Soo

    2011-01-01

    Highlights: → hASCs were differentiated into skeletal muscle cells by treatment with 5-azacytidine, FGF-2, and the supernatant of cultured hASCs. → Dystrophin and MyHC were expressed in late differentiation step by treatment with the supernatant of cultured hASCs. → hASCs expressing dystrophin and MyHC contributed to myotube formation during co-culture with mouse myoblast C2C12 cells. -- Abstract: Stem cell therapy for muscular dystrophies requires stem cells that are able to participate in the formation of new muscle fibers. However, the differentiation steps that are the most critical for this process are not clear. We investigated the myogenic phases of human adipose tissue-derived stem cells (hASCs) step by step and the capability of myotube formation according to the differentiation phase by cellular fusion with mouse myoblast C2C12 cells. In hASCs treated with 5-azacytidine and fibroblast growth factor-2 (FGF-2) for 1 day, the early differentiation step to express MyoD and myogenin was induced by FGF-2 treatment for 6 days. Dystrophin and myosin heavy chain (MyHC) expression was induced by hASC conditioned medium in the late differentiation step. Myotubes were observed only in hASCs undergoing the late differentiation step by cellular fusion with C2C12 cells. In contrast, hASCs that were normal or in the early stage were not involved in myotube formation. Our results indicate that stem cells expressing dystrophin and MyHC are more suitable for myotube formation by co-culture with myoblasts than normal or early differentiated stem cells expressing MyoD and myogenin.

  3. Precise correction of the dystrophin gene in duchenne muscular dystrophy patient induced pluripotent stem cells by TALEN and CRISPR-Cas9.

    Science.gov (United States)

    Li, Hongmei Lisa; Fujimoto, Naoko; Sasakawa, Noriko; Shirai, Saya; Ohkame, Tokiko; Sakuma, Tetsushi; Tanaka, Michihiro; Amano, Naoki; Watanabe, Akira; Sakurai, Hidetoshi; Yamamoto, Takashi; Yamanaka, Shinya; Hotta, Akitsu

    2015-01-13

    Duchenne muscular dystrophy (DMD) is a severe muscle-degenerative disease caused by a mutation in the dystrophin gene. Genetic correction of patient-derived induced pluripotent stem cells (iPSCs) by TALENs or CRISPR-Cas9 holds promise for DMD gene therapy; however, the safety of such nuclease treatment must be determined. Using a unique k-mer database, we systematically identified a unique target region that reduces off-target sites. To restore the dystrophin protein, we performed three correction methods (exon skipping, frameshifting, and exon knockin) in DMD-patient-derived iPSCs, and found that exon knockin was the most effective approach. We further investigated the genomic integrity by karyotyping, copy number variation array, and exome sequencing to identify clones with a minimal mutation load. Finally, we differentiated the corrected iPSCs toward skeletal muscle cells and successfully detected the expression of full-length dystrophin protein. These results provide an important framework for developing iPSC-based gene therapy for genetic disorders using programmable nucleases. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  4. Long-Term Efficacy of Systemic Multiexon Skipping Targeting Dystrophin Exons 45–55 With a Cocktail of Vivo-Morpholinos in Mdx52 Mice

    Directory of Open Access Journals (Sweden)

    Yusuke Echigoya

    2015-01-01

    Full Text Available Antisense-mediated exon skipping, which can restore the reading frame, is a most promising therapeutic approach for Duchenne muscular dystrophy. Remaining challenges include the limited applicability to patients and unclear function of truncated dystrophin proteins. Multiexon skipping targeting exons 45–55 at the mutation hotspot of the dystrophin gene could overcome both of these challenges. Previously, we described the feasibility of exons 45–55 skipping with a cocktail of Vivo-Morpholinos in vivo; however, the long-term efficacy and safety of Vivo-Morpholinos remains to be determined. In this study, we examined the efficacy and toxicity of exons 45–55 skipping by intravenous injections of 6 mg/kg 10-Vivo-Morpholino cocktail (0.6 mg/kg each vPMO every 2 weeks for 18 weeks to dystrophic exon-52 knockout (mdx52 mice. Systemic skipping of the entire exons 45–55 region was induced, and the Western blot analysis exhibited the restoration of 5–27% of normal levels of dystrophin protein in skeletal muscles, accompanied by improvements in histopathology and muscle strength. No obvious immune response and renal and hepatic toxicity were detected at the end-point of the treatment. We demonstrate our new regimen with the 10-Vivo-Morpholino cocktail is effective and safe for long-term repeated systemic administration in the dystrophic mouse model.

  5. Voluntary wheel running in dystrophin-deficient (mdx) mice: Relationships between exercise parameters and exacerbation of the dystrophic phenotype.

    Science.gov (United States)

    Smythe, Gayle M; White, Jason D

    2011-12-18

    Voluntary wheel running can potentially be used to exacerbate the disease phenotype in dystrophin-deficient mdx mice. While it has been established that voluntary wheel running is highly variable between individuals, the key parameters of wheel running that impact the most on muscle pathology have not been examined in detail. We conducted a 2-week test of voluntary wheel running by mdx mice and the impact of wheel running on disease pathology. There was significant individual variation in the average daily distance (ranging from 0.003 ± 0.005 km to 4.48 ± 0.96 km), culminating in a wide range (0.040 km to 67.24 km) of total cumulative distances run by individuals. There was also variation in the number and length of run/rest cycles per night, and the average running rate. Correlation analyses demonstrated that in the quadriceps muscle, a low number of high distance run/rest cycles was the most consistent indicator for increased tissue damage. The amount of rest time between running bouts was a key factor associated with gastrocnemius damage. These data emphasize the need for detailed analysis of individual running performance, consideration of the length of wheel exposure time, and the selection of appropriate muscle groups for analysis, when applying the use of voluntary wheel running to disease exacerbation and/or pre-clinical testing of the efficacy of therapeutic agents in the mdx mouse.

  6. Metabolic dysfunction and altered mitochondrial dynamics in the utrophin-dystrophin deficient mouse model of duchenne muscular dystrophy.

    Directory of Open Access Journals (Sweden)

    Meghna Pant

    Full Text Available The utrophin-dystrophin deficient (DKO mouse model has been widely used to understand the progression of Duchenne muscular dystrophy (DMD. However, it is unclear as to what extent muscle pathology affects metabolism. Therefore, the present study was focused on understanding energy expenditure in the whole animal and in isolated extensor digitorum longus (EDL muscle and to determine changes in metabolic enzymes. Our results show that the 8 week-old DKO mice consume higher oxygen relative to activity levels. Interestingly the EDL muscle from DKO mouse consumes higher oxygen per unit integral force, generates less force and performs better in the presence of pyruvate thus mimicking a slow twitch muscle. We also found that the expression of hexokinase 1 and pyruvate kinase M2 was upregulated several fold suggesting increased glycolytic flux. Additionally, there is a dramatic increase in dynamin-related protein 1 (Drp 1 and mitofusin 2 protein levels suggesting increased mitochondrial fission and fusion, a feature associated with increased energy demand and altered mitochondrial dynamics. Collectively our studies point out that the dystrophic disease has caused significant changes in muscle metabolism. To meet the increased energetic demand, upregulation of metabolic enzymes and regulators of mitochondrial fusion and fission is observed in the dystrophic muscle. A better understanding of the metabolic demands and the accompanied alterations in the dystrophic muscle can help us design improved intervention therapies along with existing drug treatments for the DMD patients.

  7. A transcriptome-based assessment of the astrocytic dystrophin-associated complex in the developing human brain.

    Science.gov (United States)

    Simon, Matthew J; Murchison, Charles; Iliff, Jeffrey J

    2018-02-01

    Astrocytes play a critical role in regulating the interface between the cerebral vasculature and the central nervous system. Contributing to this is the astrocytic endfoot domain, a specialized structure that ensheathes the entirety of the vasculature and mediates signaling between endothelial cells, pericytes, and neurons. The astrocytic endfoot has been implicated as a critical element of the glymphatic pathway, and changes in protein expression profiles in this cellular domain are linked to Alzheimer's disease pathology. Despite this, basic physiological properties of this structure remain poorly understood including the developmental timing of its formation, and the protein components that localize there to mediate its functions. Here we use human transcriptome data from male and female subjects across several developmental stages and brain regions to characterize the gene expression profile of the dystrophin-associated complex (DAC), a known structural component of the astrocytic endfoot that supports perivascular localization of the astroglial water channel aquaporin-4. Transcriptomic profiling is also used to define genes exhibiting parallel expression profiles to DAC elements, generating a pool of candidate genes that encode gene products that may contribute to the physiological function of the perivascular astrocytic endfoot domain. We found that several genes encoding transporter proteins are transcriptionally associated with DAC genes. © 2017 Wiley Periodicals, Inc.

  8. The Role of Nanobiotechnology in the Study of Dystrophin and B-Dystroglycan in Membrane Stability of Aging Skeletal Muscles

    Science.gov (United States)

    Vaseashta, Ashok

    2005-03-01

    Duchene muscular dystrophy (DMD) is one of nine types of muscular dystrophy, a group of genetic degenerative diseases, primarily affecting voluntary muscles, caused by absence of dystrophin. New experiments on mice with DMD has shown that gene therapy can reverse some symptoms of the disease. The ultimate goal of gene therapy for muscle diseases is improvement of strength and function, which will require treatment in multiple muscles simultaneously. A major limitation to gene therapy until now has been that no one had found a method by which a new gene could be delivered to all the muscles of an adult animal. Recent utilization of nanotechnology to life sciences has shown exciting promises in a wide range of disciplines, showing advances in the ability to manipulate, fabricate and alter tiny subjects at the nanometer scale. In the present investigation, we have employed such techniques to study single motors such as myosin and kinesin, as well elastic proteins viz. titin and nebulin, muscle filaments, cytoskeletal filaments, and receptors in cellular membranes and cellular organelles viz. myofibril, ribosome, and chromatin. Application of AFM to images and measures the elastic properties of single monomeric and oligomeric protein, genetically engineered titin, and nebulin molecules will be presented.

  9. A novel point mutation (G[sup [minus]1] to T) in a 5[prime] splice donor site of intron 13 of the dystrophin gene results in exon skipping and is responsible for Becker Muscular Dystrophy

    Energy Technology Data Exchange (ETDEWEB)

    Hagiwara, Yoko; Nishio, Hisahide; Kitoh, Yoshihiko; Takeshima, Yasuhiro; Narita, Naoko; Wada, Hiroko; Yokoyama, Mitsuhiro; Nakamura, Hajime; Matsuo, Masafumi (Kobe Univ. School of Medicine (Japan))

    1994-01-01

    The mutations in one-third of Duchenne and Becker muscular dystrophy patients remain unknown, as they do not involve gross rearrangements of the dystrophin gene. The authors now report a defect in the splicing of precursor mRNA (pre-mRNA), resulting from a maternally inherited mutation of the dystrophin gene in a patient with Becker muscular dystrophy. This defect results from a G-to-T transversion at the terminal nucleotide of exon 13, within the 5[prime] splice site of intron 13, and causes complete skipping of exon 13 during processing of dystrophin pre-mRNA. The predicted polypeptide encoded by the aberrant mRNA is a truncated dystrophin lacking 40 amino acids from the amino-proximal end of the rod domain. This is the first report of an intraexon point mutation that completely inactivates a 5[prime] splice donor site in dystrophin pre-mRNA. Analysis of the genomic context of the G[sup [minus]1]-to-T mutation at the 5[prime] splice site supports the exon-definition model of pre-mRNA splicing and contributes to the understanding of splice-site selection. 48 refs., 5 figs.

  10. Delivery of AAV2/9-microdystrophin genes incorporating helix 1 of the coiled-coil motif in the C-terminal domain of dystrophin improves muscle pathology and restores the level of α1-syntrophin and α-dystrobrevin in skeletal muscles of mdx mice.

    Science.gov (United States)

    Koo, Taeyoung; Malerba, Alberto; Athanasopoulos, Takis; Trollet, Capucine; Boldrin, Luisa; Ferry, Arnaud; Popplewell, Linda; Foster, Helen; Foster, Keith; Dickson, George

    2011-11-01

    Duchenne muscular dystrophy is a severe X-linked inherited muscle wasting disorder caused by mutations in the dystrophin gene. Adeno-associated virus (AAV) vectors have been extensively used to deliver genes efficiently for dystrophin expression in skeletal muscles. To overcome limited packaging capacity of AAV vectors (pathology of dystrophic mdx mice. However, the CT domain of dystrophin is thought to recruit part of the dystrophin-associated protein complex, which acts as a mediator of signaling between extracellular matrix and cytoskeleton in muscle fibers. In this study, we extended the ΔR4-23/ΔCT microdystrophin by incorporating helix 1 of the coiled-coil motif in the CT domain of dystrophin (MD2), which contains the α1-syntrophin and α-dystrobrevin binding sites. Intramuscular injection of AAV2/9 expressing CT domain-extended microdystrophin showed efficient dystrophin expression in tibialis anterior muscles of mdx mice. The presence of the CT domain of dystrophin in MD2 increased the recruitment of α1-syntrophin and α-dystrobrevin at the sarcolemma and significantly improved the muscle resistance to lengthening contraction-induced muscle damage in the mdx mice compared with MD1. These results suggest that the incorporation of helix 1 of the coiled-coil motif in the CT domain of dystrophin to the microdystrophins will substantially improve their efficiency in restoring muscle function in patients with Duchenne muscular dystrophy.

  11. Micro Manufacturing

    DEFF Research Database (Denmark)

    Hansen, Hans Nørgaard

    2003-01-01

    Manufacturing deals with systems that include products, processes, materials and production systems. These systems have functional requirements, constraints, design parameters and process variables. They must be decomposed in a systematic manner to achieve the best possible system performance....... If a micro manufacturing system isn’t designed rationally and correctly, it will be high-cost, unreliable, and not robust. For micro products and systems it is a continuously increasing challenge to create the operational basis for an industrial production. As the products through product development...... processes are made applicable to a large number of customers, the pressure in regard to developing production technologies that make it possible to produce the products at a reasonable price and in large numbers is growing. The micro/nano manufacturing programme at the Department of Manufacturing...

  12. Evaluation of multiplex ligation-dependent probe amplification analysis versus multiplex polymerase chain reaction assays in the detection of dystrophin gene rearrangements in an Iranian population subset

    Directory of Open Access Journals (Sweden)

    Nayereh Nouri

    2014-01-01

    Full Text Available Background: The Duchenne muscular dystrophy (DMD gene is located in the short arm of the X chromosome (Xp21. It spans 2.4 Mb of the human genomic DNA and is composed of 79 exons. Mutations in the Dystrophin gene result in DMD and Becker muscular dystrophy. In this study, the efficiency of multiplex ligation-dependent probe amplification (MLPA over multiplex polymerase chain reaction (PCR assays in an Iranian population was investigated. Materials and Methods: Multiplex PCR assays and MLPA analysis were carried out in 74 patients affected with DMD. Results: Multiplex PCR detected deletions in 51% of the patients with DMD. MLPA analysis could determine all the deletions detected by the multiplex PCR. Additionally, MLPA was able to identify one more deletion and duplication in patients without detectable mutations by multiplex PCR. Moreover, MLPA precisely determined the exact size of the deletions. Conclusion: Although MLPA analysis is more sensitive for detection of deletions and duplications in the dystrophin gene, multiplex PCR might be used for the initial analysis of the boys affected with DMD in the Iranian population as it was able to detect 95% of the rearrangements in patients with DMD.

  13. Cloning of human basic A1, a distinct 59-kDa dystrophin-associated protein encoded on chromosome 8q23-24

    Energy Technology Data Exchange (ETDEWEB)

    Ahn, A.H. [Harvard Medical School, Boston, MA (United States); Yoshida, Mikiharu; Hagiwara, Yasuko; Ozawa, Eijiro [National Institute of Neuroscience, Ogawa Higashi, Kodaira (Japan); Anderson, M.S.; Feener, C.A.; Selig, S. [Howard Hughes Medical Institute at Children`s Hospital, Boston, MA (United States); Kunkel, L.M. [Harvard Medical School, Boston, MA (United States)]|[Howard Hughes Medical Institute at Children`s Hosptial, Boston, MA (United States)

    1994-05-10

    Duchenne and Becker muscular dystrophies are caused by defects of dystrophin, which forms a part of the membrane cytoskeleton of specialized cells such as muscle. It has been previously shown that the dystrophin-associated protein A1 (59-kDa DAP) is actually a heterogeneous group of phosphorylated proteins consisting of an acidic ({alpha}-A1) and a distinct basic ({beta}-A1) component. Partial peptide sequence of the A1 complex purified from rabbit muscle permitted the design of oligonucleotide probes that were used to isolate a cDNA for one human isoform of A1. This cDNA encodes a basic A1 isoform that is distinct from the recently described syntrophins in Torpedo and mouse and is expressed in many tissues with at least five distinct mRNA species of 5.9, 4.8, 4.3, 3.1, and 1.5 kb. A comparison of the human cDNA sequence with the GenBank expressed sequence tag (EST) data base has identified a relative from human skeletal muscle, EST25263, which is probably a human homologue of the published mouse syntrophin 2. The authors have mapped the human basic component of A1 and EST25263 genes to chromosomes 8q23-24 and 16, respectively.

  14. Micro Programming

    OpenAIRE

    Spanjersberg , Herman

    2012-01-01

    International audience; In the 1970s a need arose to perform special arithmetic operations on minicomputers much more quickly than had been possible in the past. This paper tells the story of why micro programming was needed for special arithmetic operations on mini computers in the 1970s and how it was implemented. The paper tells how the laboratory in which the first experiment took place had a PDP-9 minicomputer from Digital Equipment Corporation and how the author, with several colleagues...

  15. Dual Myostatin and Dystrophin Exon Skipping by Morpholino Nucleic Acid Oligomers Conjugated to a Cell-penetrating Peptide Is a Promising Therapeutic Strategy for the Treatment of Duchenne Muscular Dystrophy

    Directory of Open Access Journals (Sweden)

    Alberto Malerba

    2012-01-01

    Full Text Available The knockdown of myostatin, a negative regulator of skeletal muscle mass may have important implications in disease conditions accompanied by muscle mass loss like cancer, HIV/AIDS, sarcopenia, muscle atrophy, and Duchenne muscular dystrophy (DMD. In DMD patients, where major muscle loss has occurred due to a lack of dystrophin, the therapeutic restoration of dystrophin expression alone in older patients may not be sufficient to restore the functionality of the muscles. We recently demonstrated that phosphorodiamidate morpholino oligomers (PMOs can be used to re-direct myostatin splicing and promote the expression of an out-of-frame transcript so reducing the amount of the synthesized myostatin protein. Furthermore, the systemic administration of the same PMO conjugated to an octaguanidine moiety (Vivo-PMO led to a significant increase in the mass of soleus muscle of treated mice. Here, we have further optimized the use of Vivo-PMO in normal mice and also tested the efficacy of the same PMO conjugated to an arginine-rich cell-penetrating peptide (B-PMO. Similar experiments conducted in mdx dystrophic mice showed that B-PMO targeting myostatin is able to significantly increase the tibialis anterior (TA muscle weight and when coadministered with a B-PMO targeting the dystrophin exon 23, it does not have a detrimental interaction. This study confirms that myostatin knockdown by exon skipping is a potential therapeutic strategy to counteract muscle wasting conditions and dual myostatin and dystrophin skipping has potential as a therapy for DMD.

  16. Persistent Dystrophin Protein Restoration 90 Days after a Course of Intraperitoneally Administered Naked 2′OMePS AON and ZM2 NP-AON Complexes in mdx Mice

    Directory of Open Access Journals (Sweden)

    Elena Bassi

    2012-01-01

    Full Text Available In Duchenne muscular dystrophy, the exon-skipping approach has obtained proof of concept in animal models, myogenic cell cultures, and following local and systemic administration in Duchenne patients. Indeed, we have previously demonstrated that low doses (7.5 mg/Kg/week of 2′-O-methyl-phosphorothioate antisense oligoribonucleotides (AONs adsorbed onto ZM2 nanoparticles provoke widespread dystrophin restoration 7 days after intraperitoneal treatment in mdx mice. In this study, we went on to test whether this dystrophin restoration was still measurable 90 days from the end of the same treatment. Interestingly, we found that both western blot and immunohistochemical analysis (up to 7% positive fibres were still able to detect dystrophin protein in the skeletal muscles of ZM2-AON-treated mice at this time, and the level of exon-23 skipping could still be assessed by RT real-time PCR (up to 10% of skipping percentage. In contrast, the protein was undetectable by western blot analysis in the skeletal muscles of mdx mice treated with an identical dose of naked AON, and the percentage of dystrophin-positive fibres and exon-23 skipping were reminiscent of those of untreated mdx mice. Our data therefore demonstrate the long-term residual efficacy of this systemic low-dose treatment and confirm the protective effect nanoparticles exert on AON molecules.

  17. Immunohistochemical alterations of dystrophin in congenital muscular dystrophy Alterações imuno-hístoquímicas da distrofina na distrofia muscular congênita

    Directory of Open Access Journals (Sweden)

    Lineu Cesar Werneck

    1995-09-01

    Full Text Available The dystrophin distribution in the plasma muscle membrane using immunohystochemistry was studied in 22 children with congenital muscular dystrophy. The dystrophin was detected by immunofluorescence in muscle biopsy through a polyclonal antibody. All the cases had patchy interruptions of the fluorescence in the plasma membrane. A large patchy interruption of the sarcolemma was found in 17 cases, small interruption in 12, and a combination of large and small patchy discontinuity in 7. Small gaps around the fiber like a rosary were found in 15 cases. The frequency of these abnormalities ranged cases from: all fibers in 5 cases, frequent in 8, occasional in 5, and rare in 4. Five cases had total absence of immunofluorescence. These results suggest that the dystrophin expression is abnormal in this group of children and that this type of abnormalities can not be differentiated from early Becker muscular dystrophy nor childhood autosomal recessive muscular dystrophy through immunohystochemistry alone.Foi estudada a distribuição da distrofina na membrana plasmática das fibras musculares em 22 crianças com distrofia muscular congênita, através de técnicas de imuno-histoquímica. A distrofina foi identificada nas biópsias musculares processadas a fresco, por técnicas de imunofluorescência utilizando anticorpos policlonais. Todos os casos tinham interrupções da imunofluorescência na membrana plasmática. Em 17 elas eram grandes, em 12 eram pequenas e em 7 eram de ambos os tipos. Fibras com interrupções pequenas e constantes, como um rosário, foram vistas em 15 casos. Essas anormalidades estavam presentes em todas as fibras em 5 casos, eram frequentes em 8, ocasionais em 5 e raras em 4. Cinco casos mostraram fibras sem distrofina. Esses dados sugerem que a expressão da distrofina é anormal nesse grupo de crianças. Essas anormalidades podem também ser encontradas em casos precoces de distrofia muscular de Becker e distrofia autoss

  18. Life-threatening Arrhythmias in a Becker Muscular Dystrophy Family due to the Duplication of Exons 3-4 of the Dystrophin Gene.

    Science.gov (United States)

    Ishizaki, Masatoshi; Fujimoto, Akiko; Ueyama, Hidetsugu; Nishida, Yasuto; Imamura, Shigehiro; Uchino, Makoto; Ando, Yukio

    2015-01-01

    We herein present a report of three patients with Becker muscular dystrophy in the same family who developed complete atrioventricular block or ventricular tachycardia with severe cardiomyopathy. Our cases became unable to walk in their teens, and were introduced to mechanical ventilation due to respiratory muscle weakness in their twenties and thirties. In all three cases, a medical device such as a permanent cardiac pacemaker or an implantable cardiac defibrillator was considered to be necessary. The duplication of exons 3-4 in the dystrophin gene was detected in two of the patients. In patients with Becker muscular dystrophy, complete atrioventricular block or ventricular tachycardia within a family has rarely been reported. Thus attention should be paid to the possibility of severe arrhythmias in the severe phenotype of Becker muscular dystrophy.

  19. Linkage disequilibria among (CA){sub n} polymorphisms in the human dystrophin gene and their implications in carrier detection and prenatal diagnosis in Duchenne and Becker musclar dystrophies

    Energy Technology Data Exchange (ETDEWEB)

    Chakraborty, R.; Zhong, Y.; Andrade, M. de [Univ. of Texas Graduate School of Biomedical Sciences, Houston, TX (United States)] [and others

    1994-06-01

    Four short tandem repeat loci, characterized by length polymorphisms of (CA){sub n} repeats, have been detected within introns 44, 45, 49, and 50 of the human dystrophin gene. The predicted heterozygosites for these loci range from 72 to 93%, and observed allele numbers range from 6 to 19 in 57 normal chromosomes, revealing their high degree of polymorphism. Evidence for significant disequilibria between the loci within introns 49 and 50 is found. These data appear to be consistent with observations of recombination frequencies between these markers and the length of the intron 44 in relation to the entire region. In addition, these four loci are collectively found to be 100% informative in carrier detection/prenatal diagnosis of Becker and Duchenne muscular dystrophies (B/DMD), whereas scoring the (CA){sub n} markers within introns 45 and 49 alone gives a 99.6% success rate. 13 refs., 4 tabs.

  20. Compensation for dystrophin-deficiency: ADAM12 overexpression in skeletal muscle results in increased alpha 7 integrin, utrophin and associated glycoproteins

    DEFF Research Database (Denmark)

    Moghadaszadeh, Behzad; Albrechtsen, Reidar; Guo, Ling T

    2003-01-01

    Mouse models for genetic diseases are among the most powerful tools available for developing and testing new treatment strategies. ADAM12 is a disintegrin and metalloprotease, previously demonstrated to significantly alleviate the pathology of mdx mice, a model for Duchenne muscular dystrophy...... in humans. More specifically ADAM12 appeared to prevent muscle cell necrosis in the mdx mice as evidenced by morphological analysis and by the reduced levels of serum creatine kinase. In the present study we demonstrated that ADAM12 may compensate for the dystrophin deficiency in mdx mice by increasing...... the expression and redistribution of several components of the muscle cell-adhesion complexes. First, we analyzed transgenic mice that overexpress ADAM12 and found mild myopathic changes and accelerated regeneration following acute injury. We then analyzed changes in gene-expression profiles in mdx/ADAM12...

  1. MicroED data collection and processing

    Energy Technology Data Exchange (ETDEWEB)

    Hattne, Johan; Reyes, Francis E.; Nannenga, Brent L.; Shi, Dan; Cruz, M. Jason de la [Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147 (United States); Leslie, Andrew G. W. [Medical Research Council Laboratory of Molecular Biology, Cambridge (United Kingdom); Gonen, Tamir, E-mail: gonent@janelia.hhmi.org [Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147 (United States)

    2015-07-01

    The collection and processing of MicroED data are presented. MicroED, a method at the intersection of X-ray crystallography and electron cryo-microscopy, has rapidly progressed by exploiting advances in both fields and has already been successfully employed to determine the atomic structures of several proteins from sub-micron-sized, three-dimensional crystals. A major limiting factor in X-ray crystallography is the requirement for large and well ordered crystals. By permitting electron diffraction patterns to be collected from much smaller crystals, or even single well ordered domains of large crystals composed of several small mosaic blocks, MicroED has the potential to overcome the limiting size requirement and enable structural studies on difficult-to-crystallize samples. This communication details the steps for sample preparation, data collection and reduction necessary to obtain refined, high-resolution, three-dimensional models by MicroED, and presents some of its unique challenges.

  2. Neuronal nitric oxide synthase-rescue of dystrophin/utrophin double knockout mice does not require nNOS localization to the cell membrane.

    Directory of Open Access Journals (Sweden)

    Michelle Wehling-Henricks

    Full Text Available Survival of dystrophin/utrophin double-knockout (dko mice was increased by muscle-specific expression of a neuronal nitric oxide synthase (nNOS transgene. Dko mice expressing the transgene (nNOS TG+/dko experienced delayed onset of mortality and increased life-span. The nNOS TG+/dko mice demonstrated a significant decrease in the concentration of CD163+, M2c macrophages that can express arginase and promote fibrosis. The decrease in M2c macrophages was associated with a significant reduction in fibrosis of heart, diaphragm and hindlimb muscles of nNOS TG+/dko mice. The nNOS transgene had no effect on the concentration of cytolytic, CD68+, M1 macrophages. Accordingly, we did not observe any change in the extent of muscle fiber lysis in the nNOS TG+/dko mice. These findings show that nNOS/NO (nitric oxide-mediated decreases in M2c macrophages lead to a reduction in the muscle fibrosis that is associated with increased mortality in mice lacking dystrophin and utrophin. Interestingly, the dramatic and beneficial effects of the nNOS transgene were not attributable to localization of nNOS protein at the cell membrane. We did not detect any nNOS protein at the sarcolemma in nNOS TG+/dko muscles. This important observation shows that sarcolemmal localization is not necessary for nNOS to have beneficial effects in dystrophic tissue and the presence of nNOS in the cytosol of dystrophic muscle fibers can ameliorate the pathology and most importantly, significantly increase life-span.

  3. Improvement of cardiac contractile function by peptide-based inhibition of NF-κB in the utrophin/dystrophin-deficient murine model of muscular dystrophy

    Directory of Open Access Journals (Sweden)

    Guttridge Denis C

    2011-05-01

    Full Text Available Abstract Background Duchenne muscular dystrophy (DMD is an inherited and progressive disease causing striated muscle deterioration. Patients in their twenties generally die from either respiratory or cardiac failure. In order to improve the lifespan and quality of life of DMD patients, it is important to prevent or reverse the progressive loss of contractile function of the heart. Recent studies by our labs have shown that the peptide NBD (Nemo Binding Domain, targeted at blunting Nuclear Factor κB (NF-κB signaling, reduces inflammation, enhances myofiber regeneration, and improves contractile deficits in the diaphragm in dystrophin-deficient mdx mice. Methods To assess whether cardiac function in addition to diaphragm function can be improved, we investigated physiological and histological parameters of cardiac muscle in mice deficient for both dystrophin and its homolog utrophin (double knockout = dko mice treated with NBD peptide. These dko mice show classic pathophysiological hallmarks of heart failure, including myocyte degeneration, an impaired force-frequency response and a severely blunted β-adrenergic response. Cardiac contractile function at baseline and frequencies and pre-loads throughout the in vivo range as well as β-adrenergic reserve was measured in isolated cardiac muscle preparations. In addition, we studied histopathological and inflammatory markers in these mice. Results At baseline conditions, active force development in cardiac muscles from NBD treated dko mice was more than double that of vehicle-treated dko mice. NBD treatment also significantly improved frequency-dependent behavior of the muscles. The increase in force in NBD-treated dko muscles to β-adrenergic stimulation was robustly restored compared to vehicle-treated mice. However, histological features, including collagen content and inflammatory markers were not significantly different between NBD-treated and vehicle-treated dko mice. Conclusions We conclude

  4. Deletion of Galgt2 (B4Galnt2) reduces muscle growth in response to acute injury and increases muscle inflammation and pathology in dystrophin-deficient mice.

    Science.gov (United States)

    Xu, Rui; Singhal, Neha; Serinagaoglu, Yelda; Chandrasekharan, Kumaran; Joshi, Mandar; Bauer, John A; Janssen, Paulus M L; Martin, Paul T

    2015-10-01

    Transgenic overexpression of Galgt2 (official name B4Galnt2) in skeletal muscle stimulates the glycosylation of α dystroglycan (αDG) and the up-regulation of laminin α2 and dystrophin surrogates known to inhibit muscle pathology in mouse models of congenital muscular dystrophy 1A and Duchenne muscular dystrophy. Skeletal muscle Galgt2 gene expression is also normally increased in the mdx mouse model of Duchenne muscular dystrophy compared with the wild-type mice. To assess whether this increased endogenous Galgt2 expression could affect disease, we quantified muscular dystrophy measures in mdx mice deleted for Galgt2 (Galgt2(-/-)mdx). Galgt2(-/-) mdx mice had increased heart and skeletal muscle pathology and inflammation, and also worsened cardiac function, relative to age-matched mdx mice. Deletion of Galgt2 in wild-type mice also slowed skeletal muscle growth in response to acute muscle injury. In each instance where Galgt2 expression was elevated (developing muscle, regenerating muscle, and dystrophic muscle), Galgt2-dependent glycosylation of αDG was also increased. Overexpression of Galgt2 failed to inhibit skeletal muscle pathology in dystroglycan-deficient muscles, in contrast to previous studies in dystrophin-deficient mdx muscles. This study demonstrates that Galgt2 gene expression and glycosylation of αDG are dynamically regulated in muscle and that endogenous Galgt2 gene expression can ameliorate the extent of muscle pathology, inflammation, and dysfunction in mdx mice. Copyright © 2015 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

  5. Micro club

    CERN Multimedia

    Micro club

    2014-01-01

    Opération NEMO   Pour finir en beauté les activités spéciales que le CMC a réalisé pendant cette année 2014, pour commémorer le 60ème anniversaire du CERN, et le 30ème du Micro Club, l’ Opération NEMO aura cette année un caractère très particulier. Nous allons proposer 6 fabricants de premier ordre qui offriront chacun deux ou trois produits à des prix exceptionnels. L’opération débute le lundi 17 novembre 2014. Elle se poursuivra  jusqu’au samedi 6 décembre inclus. Les délais de livraison seront de deux à trois semaines, selon les fabricants. Donc les commandes faites la dernière semaine, du 1 au 6 décembre, risquent d’arriver qu'au début du mois de janvier 2015. Liste de fabricants part...

  6. Micro club

    CERN Multimedia

    Micro Club

    2014-01-01

    Jeudi 18 septembre 2014 à 18h30 au Bât. 567 R-029 Le CERN MICRO CLUB organise un Atelier sur la sécurité informatique. La Cyber-sécurité : Ce qui se passe vraiment, comment ne pas en être victime ! Orateur : Sebastian Lopienski Adjoint au Computer Security Officer du Département IT. Sujet : Cet exposé vous présentera les modes de sécurité actuels et les problèmes touchants les applications logicielles des ordinateurs, les réseaux ainsi que leurs utilisateurs. Cela inclus des informations sur les nouveaux types de vulnérabilité, les vecteurs d'attaque récents et une vue d'ensemble sur le monde de la cyber-sécurité en 2014. Biographie : Sebastian Lopienski travaille au CERN depuis 2001. Il est actuellement adjoint au Computer Security Officer et s'occupe de la protection de...

  7. Generation of induced pluripotent stem cells from a Becker muscular dystrophy patient carrying a deletion of exons 45-55 of the dystrophin gene (CCMi002BMD-A-9 ∆45-55

    Directory of Open Access Journals (Sweden)

    Aoife Gowran

    2018-04-01

    Full Text Available Becker muscular dystrophy (BMD is a dystrophinopathy caused by mutations in the dystrophin gene on chromosome Xp21. BMD mutations result in truncated semi-functional dystrophin isoforms. Consequently, less severe clinical symptoms become apparent later in life compared to Duchenne muscular dystrophy. Dermal fibroblasts from a BMD patient were electroporated with episomal plasmids containing reprogramming factors to create the induced pluripotent stem cell line: CCMi002BMD-A-9 that showed pluripotent markers, were karyotypically normal and capable of trilineage differentiation. MLPA analyses performed on DNA extracted from CCMi002BMD-A-9 showed an in-frame deletion of exons 45 to 55 (CCMi002BMD-A-9 Δ45-55.

  8. Generation of induced pluripotent stem cells from a Becker muscular dystrophy patient carrying a deletion of exons 45-55 of the dystrophin gene (CCMi002BMD-A-9 ∆45-55).

    Science.gov (United States)

    Gowran, Aoife; Spaltro, Gabriella; Casalnuovo, Federica; Vigorelli, Vera; Spinelli, Pietro; Castiglioni, Elisa; Rovina, Davide; Paganini, Stefania; Di Segni, Marina; Gervasini, Cristina; Nigro, Patrizia; Pompilio, Giulio

    2018-04-01

    Becker muscular dystrophy (BMD) is a dystrophinopathy caused by mutations in the dystrophin gene on chromosome Xp21. BMD mutations result in truncated semi-functional dystrophin isoforms. Consequently, less severe clinical symptoms become apparent later in life compared to Duchenne muscular dystrophy. Dermal fibroblasts from a BMD patient were electroporated with episomal plasmids containing reprogramming factors to create the induced pluripotent stem cell line: CCMi002BMD-A-9 that showed pluripotent markers, were karyotypically normal and capable of trilineage differentiation. MLPA analyses performed on DNA extracted from CCMi002BMD-A-9 showed an in-frame deletion of exons 45 to 55 (CCMi002BMD-A-9 Δ45-55). Copyright © 2018 The Author(s). Published by Elsevier B.V. All rights reserved.

  9. Missense mutation Lys18Asn in dystrophin that triggers X-linked dilated cardiomyopathy decreases protein stability, increases protein unfolding, and perturbs protein structure, but does not affect protein function.

    Directory of Open Access Journals (Sweden)

    Surinder M Singh

    Full Text Available Genetic mutations in a vital muscle protein dystrophin trigger X-linked dilated cardiomyopathy (XLDCM. However, disease mechanisms at the fundamental protein level are not understood. Such molecular knowledge is essential for developing therapies for XLDCM. Our main objective is to understand the effect of disease-causing mutations on the structure and function of dystrophin. This study is on a missense mutation K18N. The K18N mutation occurs in the N-terminal actin binding domain (N-ABD. We created and expressed the wild-type (WT N-ABD and its K18N mutant, and purified to homogeneity. Reversible folding experiments demonstrated that both mutant and WT did not aggregate upon refolding. Mutation did not affect the protein's overall secondary structure, as indicated by no changes in circular dichroism of the protein. However, the mutant is thermodynamically less stable than the WT (denaturant melts, and unfolds faster than the WT (stopped-flow kinetics. Despite having global secondary structure similar to that of the WT, mutant showed significant local structural changes at many amino acids when compared with the WT (heteronuclear NMR experiments. These structural changes indicate that the effect of mutation is propagated over long distances in the protein structure. Contrary to these structural and stability changes, the mutant had no significant effect on the actin-binding function as evident from co-sedimentation and depolymerization assays. These results summarize that the K18N mutation decreases thermodynamic stability, accelerates unfolding, perturbs protein structure, but does not affect the function. Therefore, K18N is a stability defect rather than a functional defect. Decrease in stability and increase in unfolding decrease the net population of dystrophin molecules available for function, which might trigger XLDCM. Consistently, XLDCM patients have decreased levels of dystrophin in cardiac muscle.

  10. Characterization of a novel Dp71 dystrophin-associated protein complex (DAPC) present in the nucleus of HeLa cells: Members of the nuclear DAPC associate with the nuclear matrix

    International Nuclear Information System (INIS)

    Fuentes-Mera, Lizeth; Rodriguez-Munoz, Rafael; Gonzalez-Ramirez, Ricardo; Garcia-Sierra, Francisco; Gonzalez, Everardo; Mornet, Dominique; Cisneros, Bulmaro

    2006-01-01

    Dystrophin is an essential component in the assembly and maintenance of the dystrophin-associated protein complex (DAPC), which includes members of the dystroglycan, syntrophin, sarcoglycan and dystrobrevin protein families. Distinctive complexes have been described in the cell membrane of different tissues and cultured cells. In this work, we report the identification and characterization of a novel DAPC present in the nuclei of HeLa cells, which contains dystrophin Dp71 as a key component. Using confocal microscopy and cell fractionation analyses, we found the presence of Dp71, β-sarcoglycan, β-dystroglycan, α- and β-syntrophin, α1- and β-dystrobrevin and nNOS in the nuclei of HeLa cells. Furthermore, we demonstrated by co-immunoprecipitation experiments that most of these proteins form a complex in the nuclear compartment. Next, we analyze the possible association of the nuclear DAPC with the nuclear matrix. We found the presence of Dp71, β-dystroglycan, nNOS, β-sarcoglycan, α/β syntrophin, α1-dystrobrevin and β-dystrobrevin in the nuclear matrix protein fractions and in situ nuclear matrix preparations from HeLa cells. Moreover, we found that Dp71, β-dystroglycan and β-dystrobrevin co-immunoprecipitated with the nuclear matrix proteins lamin B1 and actin. The association of members of the nuclear DAPC with the nuclear matrix indicates that they may work as scaffolding proteins involved in nuclear architecture

  11. Clinical and molecular characterization of a cohort of patients with novel nucleotide alterations of the Dystrophin gene detected by direct sequencing

    Directory of Open Access Journals (Sweden)

    Corti Stefania

    2011-03-01

    Full Text Available Abstract Background Duchenne and Becker Muscular dystrophies (DMD/BMD are allelic disorders caused by mutations in the dystrophin gene, which encodes a sarcolemmal protein responsible for muscle integrity. Deletions and duplications account for approximately 75% of mutations in DMD and 85% in BMD. The implementation of techniques allowing complete gene sequencing has focused attention on small point mutations and other mechanisms underlying complex rearrangements. Methods We selected 47 patients (41 families; 35 DMD, 6 BMD without deletions and duplications in DMD gene (excluded by multiplex ligation-dependent probe amplification and multiplex polymerase chain reaction analysis. This cohort was investigated by systematic direct sequence analysis to study sequence variation. We focused our attention on rare mutational events which were further studied through transcript analysis. Results We identified 40 different nucleotide alterations in DMD gene and their clinical correlates; altogether, 16 mutations were novel. DMD probands carried 9 microinsertions/microdeletions, 19 nonsense mutations, and 7 splice-site mutations. BMD patients carried 2 nonsense mutations, 2 splice-site mutations, 1 missense substitution, and 1 single base insertion. The most frequent stop codon was TGA (n = 10 patients, followed by TAG (n = 7 and TAA (n = 4. We also analyzed the molecular mechanisms of five rare mutational events. They are two frame-shifting mutations in the DMD gene 3'end in BMD and three novel splicing defects: IVS42: c.6118-3C>A, which causes a leaky splice-site; c.9560A>G, which determines a cryptic splice-site activation and c.9564-426 T>G, which creates pseudoexon retention within IVS65. Conclusion The analysis of our patients' sample, carrying point mutations or complex rearrangements in DMD gene, contributes to the knowledge on phenotypic correlations in dystrophinopatic patients and can provide a better understanding of pre-mRNA maturation defects

  12. Black bear parathyroid hormone has greater anabolic effects on trabecular bone in dystrophin-deficient mice than in wild type mice.

    Science.gov (United States)

    Gray, Sarah K; McGee-Lawrence, Meghan E; Sanders, Jennifer L; Condon, Keith W; Tsai, Chung-Jui; Donahue, Seth W

    2012-09-01

    Duchenne muscular dystrophy (DMD) is an X-linked neuromuscular disease that has deleterious consequences in muscle and bone, leading to decreased mobility, progressive osteoporosis, and premature death. Patients with DMD experience a higher-than-average fracture rate, particularly in the proximal and distal femur and proximal tibia. The dystrophin-deficient mdx mouse is a model of DMD that demonstrates muscle degeneration and fibrosis and osteoporosis. Parathyroid hormone, an effective anabolic agent for post-menopausal and glucocorticoid-induced osteoporosis, has not been explored for DMD. Black bear parathyroid hormone (bbPTH) has been implicated in the maintenance of bone properties during extended periods of disuse (hibernation). We cloned bbPTH and found 9 amino acid residue differences from human PTH. Apoptosis was mitigated and cAMP was activated by bbPTH in osteoblast cultures. We administered 28nmol/kg of bbPTH 1-84 to 4-week old male mdx and wild type mice via daily (5×/week) subcutaneous injection for 6 weeks. Vehicle-treated mdx mice had 44% lower trabecular bone volume fraction than wild type mice. No changes were found in femoral cortical bone geometry or mechanical properties with bbPTH treatment in wild type mice, and only medio-lateral moment of inertia changed with bbPTH treatment in mdx femurs. However, μCT analyses of the trabecular regions of the distal femur and proximal tibia showed marked increases in bone volume fraction with bbPTH treatment, with a greater anabolic response (7-fold increase) in mdx mice than wild type mice (2-fold increase). Trabecular number increased in mdx long bone, but not wild type bone. Additionally, greater osteoblast area and decreased osteoclast area were observed with bbPTH treatment in mdx mice. The heightened response to PTH in mdx bone compared to wild type suggests a link between dystrophin deficiency, altered calcium signaling, and bone. These findings support further investigation of PTH as an anabolic

  13. Dramatic elevation in urinary amino terminal titin fragment excretion quantified by immunoassay in Duchenne muscular dystrophy patients and in dystrophin deficient rodents.

    Science.gov (United States)

    Robertson, Alan S; Majchrzak, Mark J; Smith, Courtney M; Gagnon, Robert C; Devidze, Nino; Banks, Glen B; Little, Sean C; Nabbie, Fizal; Bounous, Denise I; DiPiero, Janet; Jacobsen, Leslie K; Bristow, Linda J; Ahlijanian, Michael K; Stimpson, Stephen A

    2017-07-01

    Enzyme-linked and electrochemiluminescence immunoassays were developed for quantification of amino (N-) terminal fragments of the skeletal muscle protein titin (N-ter titin) and qualified for use in detection of urinary N-ter titin excretion. Urine from normal subjects contained a small but measurable level of N-ter titin (1.0 ± 0.4 ng/ml). A 365-fold increase (365.4 ± 65.0, P = 0.0001) in urinary N-ter titin excretion was seen in Duchene muscular dystrophy (DMD) patients. Urinary N-ter titin was also evaluated in dystrophin deficient rodent models. Mdx mice exhibited low urinary N-ter titin levels at 2 weeks of age followed by a robust and sustained elevation starting at 3 weeks of age, coincident with the development of systemic skeletal muscle damage in this model; fold elevation could not be determined because urinary N-ter titin was not detected in age-matched wild type mice. Levels of serum creatine kinase and serum skeletal muscle troponin I (TnI) were also low at 2 weeks, elevated at later time points and were significantly correlated with urinary N-ter titin excretion in mdx mice. Corticosteroid treatment of mdx mice resulted in improved exercise performance and lowering of both urinary N-ter titin and serum skeletal muscle TnI concentrations. Low urinary N-ter titin levels were detected in wild type rats (3.0 ± 0.6 ng/ml), while Dmd mdx rats exhibited a 556-fold increase (1652.5 ± 405.7 ng/ml, P = 0.002) (both at 5 months of age). These results suggest that urinary N-ter titin is present at low basal concentrations in normal urine and increases dramatically coincident with muscle damage produced by dystrophin deficiency. Urinary N-ter titin has potential as a facile, non-invasive and translational biomarker for DMD. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  14. Micro robot bible

    International Nuclear Information System (INIS)

    Yoon, Jin Yeong

    2000-08-01

    This book deals with micro robot, which tells of summary of robots like entertainment robots and definition of robots, introduction of micro mouse about history, composition and rules, summary of micro controller with its history, appearance and composition, introduction of stepping motor about types, structure, basic characteristics, and driving ways, summary of sensor section, power, understanding of 80C196KC micro controller, basic driving program searching a maze algorithm, smooth turn and making of tracer line.

  15. Micro intelligence robot

    International Nuclear Information System (INIS)

    Jeon, Yon Ho

    1991-07-01

    This book gives descriptions of micro robot about conception of robots and micro robot, match rules of conference of micro robots, search methods of mazes, and future and prospect of robots. It also explains making and design of 8 beat robot like making technique, software, sensor board circuit, and stepping motor catalog, speedy 3, Mr. Black and Mr. White, making and design of 16 beat robot, such as micro robot artist, Jerry 2 and magic art of shortening distances algorithm of robot simulation.

  16. Micro robot bible

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Jin Yeong

    2000-08-15

    This book deals with micro robot, which tells of summary of robots like entertainment robots and definition of robots, introduction of micro mouse about history, composition and rules, summary of micro controller with its history, appearance and composition, introduction of stepping motor about types, structure, basic characteristics, and driving ways, summary of sensor section, power, understanding of 80C196KC micro controller, basic driving program searching a maze algorithm, smooth turn and making of tracer line.

  17. Early-progressive dilated cardiomyopathy in a family with Becker muscular dystrophy related to a novel frameshift mutation in the dystrophin gene exon 27.

    Science.gov (United States)

    Tsuda, Takeshi; Fitzgerald, Kristi; Scavena, Mena; Gidding, Samuel; Cox, Mary O; Marks, Harold; Flanigan, Kevin M; Moore, Steven A

    2015-03-01

    We report a family in which two male siblings with Becker muscular dystrophy (BMD) developed severe dilated cardiomyopathy (DCM) and progressive heart failure (HF) at age 11 years; one died at age 14 years while awaiting heart transplant and the other underwent left ventricular assist device implantation at the same age. Genetic analysis of one sibling showed a novel frameshift mutation in exon 27 of Duchenne muscular dystrophy (DMD) gene (c.3779_3785delCTTTGGAinsGG), in which seven base pairs are deleted and two are inserted. Although this predicts an amino-acid substitution and premature termination (p.Thr1260Argfs*8), muscle biopsy dystrophin immunostaining instead indicates that the mutation is more likely to alter splicing. Despite relatively preserved skeletal muscular performance, both the siblings developed progressive HF secondary to early-onset DCM. In addition, their 7-year-old nephew with delayed gross motor development, mild proximal muscle weakness and markedly elevated serum creatine kinase level (>13 000 IU l(-1)) at 16 months was recently demonstrated to have the familial DMD mutation. Here, we report a novel genotype of BMD with early-onset DCM and progressive lethal HF during early adolescence.

  18. A rare subclinical or mild type of Becker muscular dystrophy caused by a single exon 48 deletion of the dystrophin gene.

    Science.gov (United States)

    Zimowski, Janusz G; Pilch, Jacek; Pawelec, Magdalena; Purzycka, Joanna K; Kubalska, Jolanta; Ziora-Jakutowicz, Karolina; Dudzińska, Magdalena; Zaremba, Jacek

    2017-08-01

    In the material of 227 families with Becker muscular dystrophy (BMD), we found nine non-consanguineous families with 17 male individuals carrying a rare mutation-a single exon 48 deletion of the dystrophin gene-who were affected with a very mild or subclinical form of BMD. They were usually detected thanks to accidental findings of elevated serum creatine phosphokinase (sCPK). A thorough clinical analysis of the carriers, both children (12) and adults (5), revealed in some of them muscle hypotonia (10/17) and/or very mild muscle weakness (9/17), as well as decreased tendon reflexes (6/17). Adults, apart from very mild muscle weakness and calf hypertrophy in some, had no significant abnormalities on neurological assessments and had good exercise tolerance. Parents of the children carriers of the exon 48 deletion are usually unaware of their children being affected, and possibly at risk of developing life-threatening cardiomyopathy. The same concerns the adult male carriers. Therefore, the authors postulate undertaking preventive measures such as cascade screening of the relatives of the probands. Newborn screening programmes of Duchenne muscular dystrophy (DMD)/BMD based on sCPK marked increase may be considered.

  19. Pregnancy after preimplantation diagnosis for a deletion in the dystrophin gene by polymerase chain reaction in embryos obtained after intracytoplasmic sperm injection

    Energy Technology Data Exchange (ETDEWEB)

    Lissens, W.; Liu, J.; Van Broeckhoven, C. [University Hospital, Brussels (Belgium)] [and others

    1994-09-01

    Duchenne muscular dystrophy (DMD) is one of the most common X-linked recessive diseases. In order to be able to perform a DMD-specific preimplantation diagnosis (PID) in a female carrier of a deletion of exons 3 to 18 in the dystrophin gene, we have developed a PCR assay to detect the deletion based on sequences of exon 17. The efficiency of this PCR was evaluated on 50 single blastomeres from 12 normal control embryos and on 41 blastomeres for 9 male and 3 female embryos from the female DMD carrier, obtained after a first preimplantation diagnosis by sexing. The exon 17 region was amplified with 100% efficiency, except in all 21 blastomeres from 6 male embryos from the carrier where no PCR signals were observed. The negative results in these blastomeres were interpreted as being found only in male embryos carrying the deletion. Intracytoplasmic sperm injection was carried out on the carrier`s metaphase II oocytes retrieved after ovarian stimulation. Embryos were analyzed for the presence of exon 17 and 2 male embryos were found to be deleted, while 4 embryos showed normal amplification signals. Three of the latter embryos were replaced, resulting in a singleton pregnancy. Amniotic cell analysis showed a normal female karyotype and DNA analysis indicated a non-carrier.

  20. Micro rapid prototyping system for micro components

    International Nuclear Information System (INIS)

    Li Xiaochun; Choi Hongseok; Yang Yong

    2002-01-01

    Similarities between silicon-based micro-electro-mechanical systems (MEMS) and Shape Deposition Manufacturing (SDM) processes are obvious: both integrate additive and subtractive processes and use part and sacrificial materials to obtain functional structures. These MEMS techniques are two-dimensional (2-D) processes for a limited number of materials while SDM enables the building of parts that have traditionally been impossible to fabricate because of their complex shapes or of their variety in materials. This work presents initial results on the development of a micro rapid prototyping system that adapts SDM methodology to micro-fabrication. This system is designed to incorporate microdeposition and laser micromachining. In the hope of obtaining a precise microdeposition, an ultrasonic-based micro powder-feeding mechanism was developed in order to form thin patterns of dry powders that can be cladded or sintered onto a substrate by a micro-sized laser beam. Furthermore, experimental results on laser micromachining using a laser beam with a wavelength of 355 nm are also presented. After further improvement, the developed micro manufacturing system could take computer-aided design (CAD) output to reproduce 3-D heterogeneous micro-components from a wide selection of materials

  1. Micro-turbines

    International Nuclear Information System (INIS)

    Tashevski, Done

    2003-01-01

    In this paper a principle of micro-turbines operation, type of micro-turbines and their characteristics is presented. It is shown their usage in cogeneration and three generation application with the characteristics, the influence of more factors on micro-turbines operation as well as the possibility for application in Macedonia. The paper is result of the author's participation in the training program 'Micro-turbine technology' in Florida, USA. The characteristics of different types micro-turbines by several world producers are shown, with accent on US micro-turbines producers (Capstone, Elliott). By using the gathered Author's knowledge, contacts and the previous knowledge, conclusions and recommendations for implementation of micro-turbines in Macedonia are given. (Author)

  2. A comparative study of N-glycolylneuraminic acid (Neu5Gc and cytotoxic T cell (CT carbohydrate expression in normal and dystrophin-deficient dog and human skeletal muscle.

    Directory of Open Access Journals (Sweden)

    Paul T Martin

    Full Text Available The expression of N-glycolylneuraminic acid (Neu5Gc and the cytotoxic T cell (CT carbohydrate can impact the severity of muscular dystrophy arising from the loss of dystrophin in mdx mice. Here, we describe the expression of these two glycans in skeletal muscles of dogs and humans with or without dystrophin-deficiency. Neu5Gc expression was highly reduced (>95% in muscle from normal golden retriever crosses (GR, n = 3 and from golden retriever with muscular dystrophy (GRMD, n = 5 dogs at multiple ages (3, 6 and 13 months when compared to mouse muscle, however, overall sialic acid expression in GR and GRMD muscles remained high at all ages. Neu5Gc was expressed on only a minority of GRMD satellite cells, CD8⁺ T lymphocytes and macrophages. Human muscle from normal (no evident disease, n = 3, Becker (BMD, n = 3 and Duchenne (DMD, n = 3 muscular dystrophy individuals had absent to very low Neu5Gc staining, but some punctate intracellular muscle staining was present in BMD and DMD muscles. The CT carbohydrate was localized to the neuromuscular junction in GR muscle, while GRMD muscles had increased expression on a subset of myofibers and macrophages. In humans, the CT carbohydrate was ectopically expressed on the sarcolemmal membrane of some BMD muscles, but not normal human or DMD muscles. These data are consistent with the notion that altered Neu5Gc and CT carbohydrate expression may modify disease severity resulting from dystrophin deficiency in dogs and humans.

  3. Micro-propulsion and micro-combustion; Micropropulsion microcombustion

    Energy Technology Data Exchange (ETDEWEB)

    Ribaud, Y.; Dessornes, O.

    2002-10-01

    The AAAF (french space and aeronautic association) organized at Paris a presentation on the micro-propulsion. The first part was devoted to the thermal micro-machines for micro drones, the second part to the micro-combustion applied to micro-turbines. (A.L.B.)

  4. Micro-Organ Device

    Science.gov (United States)

    Gonda, Steve R. (Inventor); Chang, Robert C. (Inventor); Starly, Binil (Inventor); Culbertson, Christopher (Inventor); Holtorf, Heidi L. (Inventor); Sun, Wei (Inventor); Leslie, Julia (Inventor)

    2013-01-01

    A method for fabricating a micro-organ device comprises providing a microscale support having one or more microfluidic channels and one or more micro-chambers for housing a micro-organ and printing a micro-organ on the microscale support using a cell suspension in a syringe controlled by a computer-aided tissue engineering system, wherein the cell suspension comprises cells suspended in a solution containing a material that functions as a three-dimensional scaffold. The printing is performed with the computer-aided tissue engineering system according to a particular pattern. The micro-organ device comprises at least one micro-chamber each housing a micro-organ; and at least one microfluidic channel connected to the micro-chamber, wherein the micro-organ comprises cells arranged in a configuration that includes microscale spacing between portions of the cells to facilitate diffusion exchange between the cells and a medium supplied from the at least one microfluidic channel.

  5. 'Micro-8' micro-computer system

    International Nuclear Information System (INIS)

    Yagi, Hideyuki; Nakahara, Yoshinori; Yamada, Takayuki; Takeuchi, Norio; Koyama, Kinji

    1978-08-01

    The micro-computer Micro-8 system has been developed to organize a data exchange network between various instruments and a computer group including a large computer system. Used for packet exchangers and terminal controllers, the system consists of ten kinds of standard boards including a CPU board with INTEL-8080 one-chip-processor. CPU architecture, BUS architecture, interrupt control, and standard-boards function are explained in circuit block diagrams. Operations of the basic I/O device, digital I/O board and communication adapter are described with definitions of the interrupt ramp status, I/O command, I/O mask, data register, etc. In the appendixes are circuit drawings, INTEL-8080 micro-processor specifications, BUS connections, I/O address mappings, jumper connections of address selection, and interface connections. (author)

  6. Search for Bs0 --> micro+ micro- and B0 --> micro+ micro- decays with 2 fb-1 of pp collisions.

    Science.gov (United States)

    Aaltonen, T; Adelman, J; Akimoto, T; Albrow, M G; Alvarez González, B; Amerio, S; Amidei, D; Anastassov, A; Annovi, A; Antos, J; Aoki, M; Apollinari, G; Apresyan, A; Arisawa, T; Artikov, A; Ashmanskas, W; Attal, A; Aurisano, A; Azfar, F; Azzi-Bacchetta, P; Azzurri, P; Bacchetta, N; Badgett, W; Barbaro-Galtieri, A; Barnes, V E; Barnett, B A; Baroiant, S; Bartsch, V; Bauer, G; Beauchemin, P-H; Bedeschi, F; Bednar, P; Behari, S; Bellettini, G; Bellinger, J; Belloni, A; Benjamin, D; Beretvas, A; Beringer, J; Berry, T; Bhatti, A; Binkley, M; Bisello, D; Bizjak, I; Blair, R E; Blocker, C; Blumenfeld, B; Bocci, A; Bodek, A; Boisvert, V; Bolla, G; Bolshov, A; Bortoletto, D; Boudreau, J; Boveia, A; Brau, B; Bridgeman, A; Brigliadori, L; Bromberg, C; Brubaker, E; Budagov, J; Budd, H S; Budd, S; Burkett, K; Busetto, G; Bussey, P; Buzatu, A; Byrum, K L; Cabrera, S; Campanelli, M; Campbell, M; Canelli, F; Canepa, A; Carlsmith, D; Carosi, R; Carrillo, S; Carron, S; Casal, B; Casarsa, M; Castro, A; Catastini, P; Cauz, D; Cavalli-Sforza, M; Cerri, A; Cerrito, L; Chang, S H; Chen, Y C; Chertok, M; Chiarelli, G; Chlachidze, G; Chlebana, F; Cho, K; Chokheli, D; Chou, J P; Choudalakis, G; Chuang, S H; Chung, K; Chung, W H; Chung, Y S; Ciobanu, C I; Ciocci, M A; Clark, A; Clark, D; Compostella, G; Convery, M E; Conway, J; Cooper, B; Copic, K; Cordelli, M; Cortiana, G; Crescioli, F; Cuenca Almenar, C; Cuevas, J; Culbertson, R; Cully, J C; Dagenhart, D; Datta, M; Davies, T; de Barbaro, P; De Cecco, S; Deisher, A; De Lentdecker, G; De Lorenzo, G; Dell'orso, M; Demortier, L; Deng, J; Deninno, M; De Pedis, D; Derwent, P F; Di Giovanni, G P; Dionisi, C; Di Ruzza, B; Dittmann, J R; D'Onofrio, M; Donati, S; Dong, P; Donini, J; Dorigo, T; Dube, S; Efron, J; Erbacher, R; Errede, D; Errede, S; Eusebi, R; Fang, H C; Farrington, S; Fedorko, W T; Feild, R G; Feindt, M; Fernandez, J P; Ferrazza, C; Field, R; Flanagan, G; Forrest, R; Forrester, S; Franklin, M; Freeman, J C; Furic, I; Gallinaro, M; Galyardt, J; Garberson, F; Garcia, J E; Garfinkel, A F; Genser, K; Gerberich, H; Gerdes, D; Giagu, S; Giakoumopolou, V; Giannetti, P; Gibson, K; Gimmell, J L; Ginsburg, C M; Giokaris, N; Giordani, M; Giromini, P; Giunta, M; Glagolev, V; Glenzinski, D; Gold, M; Goldschmidt, N; Golossanov, A; Gomez, G; Gomez-Ceballos, G; Goncharov, M; González, O; Gorelov, I; Goshaw, A T; Goulianos, K; Gresele, A; Grinstein, S; Grosso-Pilcher, C; Grundler, U; Guimaraes da Costa, J; Gunay-Unalan, Z; Haber, C; Hahn, K; Hahn, S R; Halkiadakis, E; Hamilton, A; Han, B-Y; Han, J Y; Handler, R; Happacher, F; Hara, K; Hare, D; Hare, M; Harper, S; Harr, R F; Harris, R M; Hartz, M; Hatakeyama, K; Hauser, J; Hays, C; Heck, M; Heijboer, A; Heinemann, B; Heinrich, J; Henderson, C; Herndon, M; Heuser, J; Hewamanage, S; Hidas, D; Hill, C S; Hirschbuehl, D; Hocker, A; Hou, S; Houlden, M; Hsu, S-C; Huffman, B T; Hughes, R E; Husemann, U; Huston, J; Incandela, J; Introzzi, G; Iori, M; Ivanov, A; Iyutin, B; James, E; Jayatilaka, B; Jeans, D; Jeon, E J; Jindariani, S; Johnson, W; Jones, M; Joo, K K; Jun, S Y; Jung, J E; Junk, T R; Kamon, T; Kar, D; Karchin, P E; Kato, Y; Kephart, R; Kerzel, U; Khotilovich, V; Kilminster, B; Kim, D H; Kim, H S; Kim, J E; Kim, M J; Kim, S B; Kim, S H; Kim, Y K; Kimura, N; Kirsch, L; Klimenko, S; Klute, M; Knuteson, B; Ko, B R; Koay, S A; Kondo, K; Kong, D J; Konigsberg, J; Korytov, A; Kotwal, A V; Kraus, J; Kreps, M; Kroll, J; Krumnack, N; Kruse, M; Krutelyov, V; Kubo, T; Kuhlmann, S E; Kuhr, T; Kulkarni, N P; Kusakabe, Y; Kwang, S; Laasanen, A T; Lai, S; Lami, S; Lammel, S; Lancaster, M; Lander, R L; Lannon, K; Lath, A; Latino, G; Lazzizzera, I; Lecompte, T; Lee, J; Lee, J; Lee, Y J; Lee, S W; Lefèvre, R; Leonardo, N; Leone, S; Levy, S; Lewis, J D; Lin, C; Lin, C S; Linacre, J; Lindgren, M; Lipeles, E; Lister, A; Litvintsev, D O; Liu, T; Lockyer, N S; Loginov, A; Loreti, M; Lovas, L; Lu, R-S; Lucchesi, D; Lueck, J; Luci, C; Lujan, P; Lukens, P; Lungu, G; Lyons, L; Lys, J; Lysak, R; Lytken, E; Mack, P; Macqueen, D; Madrak, R; Maeshima, K; Makhoul, K; Maki, T; Maksimovic, P; Malde, S; Malik, S; Manca, G; Manousakis, A; Margaroli, F; Marino, C; Marino, C P; Martin, A; Martin, M; Martin, V; Martínez, M; Martínez-Ballarín, R; Maruyama, T; Mastrandrea, P; Masubuchi, T; Mattson, M E; Mazzanti, P; McFarland, K S; McIntyre, P; McNulty, R; Mehta, A; Mehtala, P; Menzemer, S; Menzione, A; Merkel, P; Mesropian, C; Messina, A; Miao, T; Miladinovic, N; Miles, J; Miller, R; Mills, C; Milnik, M; Mitra, A; Mitselmakher, G; Miyake, H; Moed, S; Moggi, N; Moon, C S; Moore, R; Morello, M; Movilla Fernandez, P; Mülmenstädt, J; Mukherjee, A; Muller, Th; Mumford, R; Murat, P; Mussini, M; Nachtman, J; Nagai, Y; Nagano, A; Naganoma, J; Nakamura, K; Nakano, I; Napier, A; Necula, V; Neu, C; Neubauer, M S; Nielsen, J; Nodulman, L; Norman, M; Norniella, O; Nurse, E; Oh, S H; Oh, Y D; Oksuzian, I; Okusawa, T; Oldeman, R; Orava, R; Osterberg, K; Pagan Griso, S; Pagliarone, C; Palencia, E; Papadimitriou, V; Papaikonomou, A; Paramonov, A A; Parks, B; Pashapour, S; Patrick, J; Pauletta, G; Paulini, M; Paus, C; Pellett, D E; Penzo, A; Phillips, T J; Piacentino, G; Piedra, J; Pinera, L; Pitts, K; Plager, C; Pondrom, L; Portell, X; Poukhov, O; Pounder, N; Prakoshyn, F; Pronko, A; Proudfoot, J; Ptohos, F; Punzi, G; Pursley, J; Rademacker, J; Rahaman, A; Ramakrishnan, V; Ranjan, N; Redondo, I; Reisert, B; Rekovic, V; Renton, P; Rescigno, M; Richter, S; Rimondi, F; Ristori, L; Robson, A; Rodrigo, T; Rogers, E; Rolli, S; Roser, R; Rossi, M; Rossin, R; Roy, P; Ruiz, A; Russ, J; Rusu, V; Saarikko, H; Safonov, A; Sakumoto, W K; Salamanna, G; Saltó, O; Santi, L; Sarkar, S; Sartori, L; Sato, K; Savoy-Navarro, A; Scheidle, T; Schlabach, P; Schmidt, E E; Schmidt, M A; Schmidt, M P; Schmitt, M; Schwarz, T; Scodellaro, L; Scott, A L; Scribano, A; Scuri, F; Sedov, A; Seidel, S; Seiya, Y; Semenov, A; Sexton-Kennedy, L; Sfyria, A; Shalhout, S Z; Shapiro, M D; Shears, T; Shepard, P F; Sherman, D; Shimojima, M; Shochet, M; Shon, Y; Shreyber, I; Sidoti, A; Sinervo, P; Sisakyan, A; Slaughter, A J; Slaunwhite, J; Sliwa, K; Smith, J R; Snider, F D; Snihur, R; Soderberg, M; Soha, A; Somalwar, S; Sorin, V; Spalding, J; Spinella, F; Spreitzer, T; Squillacioti, P; Stanitzki, M; St Denis, R; Stelzer, B; Stelzer-Chilton, O; Stentz, D; Strologas, J; Stuart, D; Suh, J S; Sukhanov, A; Sun, H; Suslov, I; Suzuki, T; Taffard, A; Takashima, R; Takeuchi, Y; Tanaka, R; Tecchio, M; Teng, P K; Terashi, K; Thom, J; Thompson, A S; Thompson, G A; Thomson, E; Tipton, P; Tiwari, V; Tkaczyk, S; Toback, D; Tokar, S; Tollefson, K; Tomura, T; Tonelli, D; Torre, S; Torretta, D; Tourneur, S; Trischuk, W; Tu, Y; Turini, N; Ukegawa, F; Uozumi, S; Vallecorsa, S; van Remortel, N; Varganov, A; Vataga, E; Vázquez, F; Velev, G; Vellidis, C; Veszpremi, V; Vidal, M; Vidal, R; Vila, I; Vilar, R; Vine, T; Vogel, M; Volobouev, I; Volpi, G; Würthwein, F; Wagner, P; Wagner, R G; Wagner, R L; Wagner-Kuhr, J; Wagner, W; Wakisaka, T; Wallny, R; Wang, S M; Warburton, A; Waters, D; Weinberger, M; Wester, W C; Whitehouse, B; Whiteson, D; Wicklund, A B; Wicklund, E; Williams, G; Williams, H H; Wilson, P; Winer, B L; Wittich, P; Wolbers, S; Wolfe, C; Wright, T; Wu, X; Wynne, S M; Yagil, A; Yamamoto, K; Yamaoka, J; Yamashita, T; Yang, C; Yang, U K; Yang, Y C; Yao, W M; Yeh, G P; Yoh, J; Yorita, K; Yoshida, T; Yu, G B; Yu, I; Yu, S S; Yun, J C; Zanello, L; Zanetti, A; Zaw, I; Zhang, X; Zheng, Y; Zucchelli, S

    2008-03-14

    We have performed a search for B(s)(0) --> micro(+) micro(-) and B(0) --> micro(+) micro(-) decays in pp collisions at square root s = 1.96 TeV using 2 fb(-1) of integrated luminosity collected by the CDF II detector at the Fermilab Tevatron Collider. The observed number of B(s)(0) and B0 candidates is consistent with background expectations. The resulting upper limits on the branching fractions are B(B(s)0) --> micro(+) micro(-)) micro(+) micro(-))<1.8 x 10(-8) at 95% C.L.

  7. Treatment with the anti-IL-6 receptor antibody attenuates muscular dystrophy via promoting skeletal muscle regeneration in dystrophin-/utrophin-deficient mice.

    Science.gov (United States)

    Wada, Eiji; Tanihata, Jun; Iwamura, Akira; Takeda, Shin'ichi; Hayashi, Yukiko K; Matsuda, Ryoichi

    2017-10-27

    Chronic increases in the levels of the inflammatory cytokine interleukin-6 (IL-6) in serum and skeletal muscle are thought to contribute to the progression of muscular dystrophy. Dystrophin/utrophin double-knockout (dKO) mice develop a more severe and progressive muscular dystrophy than the mdx mice, the most common murine model of Duchenne muscular dystrophy (DMD). In particular, dKO mice have smaller body sizes and muscle diameters, and develop progressive kyphosis and fibrosis in skeletal and cardiac muscles. As mdx mice and DMD patients, we found that IL-6 levels in the skeletal muscle were significantly increased in dKO mice. Thus, in this study, we aimed to analyze the effects of IL-6 receptor (IL-6R) blockade on the muscle pathology of dKO mice. Male dKO mice were administered an initial injection (200 mg/kg intraperitoneally (i.p.)) of either the anti-IL-6R antibody MR16-1 or an isotype-matched control rat IgG at the age of 14 days, and were then given weekly injections (25 mg/kg i.p.) until 90 days of age. Treatment of dKO mice with the MR16-1 antibody successfully inhibited the IL-6 pathway in the skeletal muscle and resulted in a significant reduction in the expression levels of phosphorylated signal transducer and activator of transcription 3 in the skeletal muscle. Pathologically, a significant increase in the area of embryonic myosin heavy chain-positive myofibers and muscle diameter, and reduced fibrosis in the quadriceps muscle were observed. These results demonstrated the therapeutic effects of IL-6R blockade on promoting muscle regeneration. Consistently, serum creatine kinase levels were decreased. Despite these improvements observed in the limb muscles, degeneration of the diaphragm and cardiac muscles was not ameliorated by the treatment of mice with the MR16-1 antibody. As no adverse effects of treatment with the MR16-1 antibody were observed, our results indicate that the anti-IL-6R antibody is a potential therapy for muscular dystrophy

  8. Micro Elector Mechanical Systems

    International Nuclear Information System (INIS)

    Yun, Jun Bo; Jo, Il Ju; Choi, Yoon Seok

    1996-09-01

    This book consists of seven chapters, which are the flow of the age from macro world to micro world, what is MEMS, semiconductor, micro machining and MEMS, where do MEMS goes to?, How to make MEMS, MEMS in the future and knowing about MEMS more than. This book is written to explain in ease and fun. It deals with MEMS in IT, BT, NT, ST, micro robot technology, basic process for making MEMS such as Bulk micromachining, surface micromachining LGA technology, DARPA and organization in domestic and overseas and academy and journal related MEMS.

  9. Micro-educational reproduction

    DEFF Research Database (Denmark)

    Andrade, Stefan Bastholm; Thomsen, Jens Peter

    2017-01-01

    This study analyzes the persistence of educational inequality in advanced industrialized societies with expanding and differentiated education systems. Using Denmark as a case, we investigate changes in immobility patterns for cohorts born 1960–1981 and develop a new micro-educational classificat...... forms of reproduction. In addition, the micro-educational approach far better explains the immobility of sons than it explains that of daughters, revealing important gender differences in the immobility patterns for sons and daughters......., in particular for sons. We also find great variation in immobility for specific micro-educations within the university level. Studies of educational immobility would therefore benefit from paying attention to micro-educational classifications, because they capture patterns of multidimensional, disaggregated...

  10. Micro metal forming

    CERN Document Server

    2013-01-01

    Micro Metal Forming, i. e. forming of parts and features with dimensions below 1 mm, is a young area of research in the wide field of metal forming technologies, expanding the limits for applying metal forming towards micro technology. The essential challenges arise from the reduced geometrical size and the increased lot size. In order to enable potential users to apply micro metal forming in production, information about the following topics are given: tribological behavior: friction between tool and work piece as well as tool wear mechanical behavior: strength and formability of the work piece material, durability of the work pieces size effects: basic description of effects occurring due to the fact, that the quantitative relation between different features changes with decreasing size process windows and limits for forming processes tool making methods numerical modeling of processes and process chains quality assurance and metrology All topics are discussed with respect to the questions relevant to micro...

  11. Micro-mixer/combustor

    KAUST Repository

    Badra, Jihad Ahmad; Masri, Assaad Rachid

    2014-01-01

    A micro-mixer/combustor to mix fuel and oxidant streams into combustible mixtures where flames resulting from combustion of the mixture can be sustained inside its combustion chamber is provided. The present design is particularly suitable

  12. Micro-surgical endodontics.

    Science.gov (United States)

    Eliyas, S; Vere, J; Ali, Z; Harris, I

    2014-02-01

    Non-surgical endodontic retreatment is the treatment of choice for endodontically treated teeth with recurrent or residual disease in the majority of cases. In some cases, surgical endodontic treatment is indicated. Successful micro-surgical endodontic treatment depends on the accuracy of diagnosis, appropriate case selection, the quality of the surgical skills, and the application of the most appropriate haemostatic agents and biomaterials. This article describes the armamentarium and technical procedures involved in performing micro-surgical endodontics to a high standard.

  13. Micro Calorimeter for Batteries

    Energy Technology Data Exchange (ETDEWEB)

    Santhanagopalan, Shriram [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

    2017-08-01

    As battery technology forges ahead and consumer demand for safer, more affordable, high-performance batteries grows, the National Renewable Energy Laboratory (NREL) has added a patented Micro Calorimeter to its existing family of R&D 100 Award-winning Isothermal Battery Calorimeters (IBCs). The Micro Calorimeter examines the thermal signature of battery chemistries early on in the design cycle using popular coin cell and small pouch cell designs, which are simple to fabricate and study.

  14. Urban micro-grids

    International Nuclear Information System (INIS)

    Faure, Maeva; Salmon, Martin; El Fadili, Safae; Payen, Luc; Kerlero, Guillaume; Banner, Arnaud; Ehinger, Andreas; Illouz, Sebastien; Picot, Roland; Jolivet, Veronique; Michon Savarit, Jeanne; Strang, Karl Axel

    2017-02-01

    ENEA Consulting published the results of a study on urban micro-grids conducted in partnership with the Group ADP, the Group Caisse des Depots, ENEDIS, Omexom, Total and the Tuck Foundation. This study offers a vision of the definition of an urban micro-grid, the value brought by a micro-grid in different contexts based on real case studies, and the upcoming challenges that micro-grid stakeholders will face (regulation, business models, technology). The electric production and distribution system, as the backbone of an increasingly urbanized and energy dependent society, is urged to shift towards a more resilient, efficient and environment-friendly infrastructure. Decentralisation of electricity production into densely populated areas is a promising opportunity to achieve this transition. A micro-grid enhances local production through clustering electricity producers and consumers within a delimited electricity network; it has the ability to disconnect from the main grid for a limited period of time, offering an energy security service to its customers during grid outages for example. However: The islanding capability is an inherent feature of the micro-grid concept that leads to a significant premium on electricity cost, especially in a system highly reliant on intermittent electricity production. In this case, a smart grid, with local energy production and no islanding capability, can be customized to meet relevant sustainability and cost savings goals at lower costs For industrials, urban micro-grids can be economically profitable in presence of high share of reliable energy production and thermal energy demand micro-grids face strong regulatory challenges that should be overcome for further development Whether islanding is or is not implemented into the system, end-user demand for a greener, more local, cheaper and more reliable energy, as well as additional services to the grid, are strong drivers for local production and consumption. In some specific cases

  15. Barbed micro-spikes for micro-scale biopsy

    Science.gov (United States)

    Byun, Sangwon; Lim, Jung-Min; Paik, Seung-Joon; Lee, Ahra; Koo, Kyo-in; Park, Sunkil; Park, Jaehong; Choi, Byoung-Doo; Seo, Jong Mo; Kim, Kyung-ah; Chung, Hum; Song, Si Young; Jeon, Doyoung; Cho, Dongil

    2005-06-01

    Single-crystal silicon planar micro-spikes with protruding barbs are developed for micro-scale biopsy and the feasibility of using the micro-spike as a micro-scale biopsy tool is evaluated for the first time. The fabrication process utilizes a deep silicon etch to define the micro-spike outline, resulting in protruding barbs of various shapes. Shanks of the fabricated micro-spikes are 3 mm long, 100 µm thick and 250 µm wide. Barbs protruding from micro-spike shanks facilitate the biopsy procedure by tearing off and retaining samples from target tissues. Micro-spikes with barbs successfully extracted tissue samples from the small intestines of the anesthetized pig, whereas micro-spikes without barbs failed to obtain a biopsy sample. Parylene coating can be applied to improve the biocompatibility of the micro-spike without deteriorating the biopsy function of the micro-spike. In addition, to show that the biopsy with the micro-spike can be applied to tissue analysis, samples obtained by micro-spikes were examined using immunofluorescent staining. Nuclei and F-actin of cells which are extracted by the micro-spike from a transwell were clearly visualized by immunofluorescent staining.

  16. Tunable micro-optics

    CERN Document Server

    Duppé, Claudia

    2015-01-01

    Presenting state-of-the-art research into the dynamic field of tunable micro-optics, this is the first book to provide a comprehensive survey covering a varied range of topics including novel materials, actuation concepts and new imaging systems in optics. Internationally renowned researchers present a diverse range of chapters on cutting-edge materials, devices and subsystems, including soft matter, artificial muscles, tunable lenses and apertures, photonic crystals, and complete tunable imagers. Special contributions also provide in-depth treatment of micro-optical characterisation, scanners, and the use of natural eye models as inspiration for new concepts in advanced optics. With applications extending from medical diagnosis to fibre telecommunications, Tunable Micro-optics equips readers with a solid understanding of the broader technical context through its interdisciplinary approach to the realisation of new types of optical systems. This is an essential resource for engineers in industry and academia,...

  17. A novel micro wiggler

    International Nuclear Information System (INIS)

    Liu Qingxiang; Xu Yong

    1995-01-01

    A novel structure of the micro-wiggler is presented. The authors developed a simplified theoretical model of the micro-wiggler. According to the model, an analytic formula of the magnetic field in two dimensions is got. A calculated program (PWMW-I) is developed from the formula. PWMW-I can calculate the field on the axis and the off-axis for the number of periods N, and the entrance or the exit of the micro-wiggler. Three model with different period (10 mm, 5 mm and 3 mm) is designed on the program. The 5T peak field for the period of 3 mm at the gap of 1 mm is got

  18. Micro Mobility Marketing

    DEFF Research Database (Denmark)

    Hosbond, Jens Henrik; Skov, Mikael B.

    2008-01-01

    , in our case a medium-sized retail supermarket. Two prototypes based on push and pull marketing strategies are implemented and evaluated. Taking outset in a synthesis of central issues in contemporary research on mobile marketing, we discuss their role in micro mobility marketing to point to similarities......Mobile marketing refers to marketing of services or goods using mobile technology and mobile marketing holds potentially great economical opportunities. Traditionally, mobile marketing has been viewed as mobility in the large taking place virtually anywhere, anytime. Further, research shows...... considerable number of studies on push-based SMS mobile marketing campaigns. This paper explores a related yet different form of mobile marketing namely micro mobility marketing. Micro mobility marketing denotes mobility in the small, meaning that promotion of goods takes place within a circumscribed location...

  19. Methods and systems for micro machines

    Energy Technology Data Exchange (ETDEWEB)

    Stalford, Harold L.

    2018-03-06

    A micro machine may be in or less than the micrometer domain. The micro machine may include a micro actuator and a micro shaft coupled to the micro actuator. The micro shaft is operable to be driven by the micro actuator. A tool is coupled to the micro shaft and is operable to perform work in response to at least motion of the micro shaft.

  20. Micro energy harvesting

    CERN Document Server

    Briand, Danick; Roundy, Shad

    2015-01-01

    With its inclusion of the fundamentals, systems and applications, this reference provides readers with the basics of micro energy conversion along with expert knowledge on system electronics and real-life microdevices. The authors address different aspects of energy harvesting at the micro scale with a focus on miniaturized and microfabricated devices. Along the way they provide an overview of the field by compiling knowledge on the design, materials development, device realization and aspects of system integration, covering emerging technologies, as well as applications in power management, e

  1. Micro/Nano manufacturing

    DEFF Research Database (Denmark)

    Tosello, Guido

    2017-01-01

    Micro- and nano-scale manufacturing has been the subject of an increasing amount of interest and research effort worldwide in both academia and industry over the past 10 years.Traditional (MEMS) manufacturing, but also precision manufacturing technologies have been developed to cover micro......-scale dimensions and accuracies. Furthermore, these fundamentally different technology ecosystems are currently combined in order to exploit strengths of both platforms. One example is the use of lithography-based technologies to establish nanostructures that are subsequently transferred to 3D geometries via...

  2. Micro-RNAs

    DEFF Research Database (Denmark)

    Taipaleenmäki, H.; Hokland, L. B.; Chen, Li

    2012-01-01

    Osteoblast differentiation and bone formation (osteogenesis) are regulated by transcriptional and post-transcriptional mechanisms. Recently, a novel class of regulatory factors termed microRNAs has been identified as playing an important role in the regulation of many aspects of osteoblast biology...... including proliferation, differentiation, metabolism and apoptosis. Also, preliminary data from animal disease models suggest that targeting miRNAs in bone can be a novel approach to increase bone mass. This review highlights the current knowledge of microRNA biology and their role in bone formation...

  3. Lectures in Micro Meteorology

    DEFF Research Database (Denmark)

    Larsen, Søren Ejling

    This report contains the notes from my lectures on Micro scale meteorology at the Geophysics Department of the Niels Bohr Institute of Copenhagen University. In the period 1993-2012, I was responsible for this course at the University. At the start of the course, I decided that the text books...... available in meteorology at that time did not include enough of the special flavor of micro meteorology that characterized the work of the meteorology group at Risø (presently of the Institute of wind energy of the Danish Technical University). This work was focused on Boundary layer flows and turbulence...

  4. Micro-manufacturing: design and manufacturing of micro-products

    National Research Council Canada - National Science Library

    Koç, Muammer; Özel, Tuğrul

    2011-01-01

    .... After addressing the fundamentals and non-metallic-based micro-manufacturing processes in the semiconductor industry, it goes on to address specific metallic-based micro-manufacturing processes...

  5. Micro-Avionics Multi-Purpose Platform (MicroAMPP)

    Data.gov (United States)

    National Aeronautics and Space Administration — The Micro-Avionics Multi-Purpose Platform (MicroAMPP) is a common avionics architecture supporting microsatellites, launch vehicles, and upper-stage carrier...

  6. Structure of catalase determined by MicroED

    Science.gov (United States)

    Nannenga, Brent L; Shi, Dan; Hattne, Johan; Reyes, Francis E; Gonen, Tamir

    2014-01-01

    MicroED is a recently developed method that uses electron diffraction for structure determination from very small three-dimensional crystals of biological material. Previously we used a series of still diffraction patterns to determine the structure of lysozyme at 2.9 Å resolution with MicroED (Shi et al., 2013). Here we present the structure of bovine liver catalase determined from a single crystal at 3.2 Å resolution by MicroED. The data were collected by continuous rotation of the sample under constant exposure and were processed and refined using standard programs for X-ray crystallography. The ability of MicroED to determine the structure of bovine liver catalase, a protein that has long resisted atomic analysis by traditional electron crystallography, demonstrates the potential of this method for structure determination. DOI: http://dx.doi.org/10.7554/eLife.03600.001 PMID:25303172

  7. Cold Gas Micro Propulsion

    NARCIS (Netherlands)

    Louwerse, M.C.

    2009-01-01

    This thesis describes the development of a micro propulsion system. The trend of miniaturization of satellites requires small sized propulsion systems. For particular missions it is important to maintain an accurate distance between multiple satellites. Satellites drift apart due to differences in

  8. Tolerances in micro manufacturing

    DEFF Research Database (Denmark)

    Hansen, Hans Nørgaard; Zhang, Yang; Islam, Aminul

    This paper describes a method for analysis of tolerances in micro manufacturing. It proposes a mapping oftolerances to dimensions and compares this with current available international standards. The analysisdocuments that tolerances are not scaled down as the absolute dimension. In practice...

  9. Micro-Scale Thermoacoustics

    Science.gov (United States)

    Offner, Avshalom; Ramon, Guy Z.

    2016-11-01

    Thermoacoustic phenomena - conversion of heat to acoustic oscillations - may be harnessed for construction of reliable, practically maintenance-free engines and heat pumps. Specifically, miniaturization of thermoacoustic devices holds great promise for cooling of micro-electronic components. However, as devices size is pushed down to micro-meter scale it is expected that non-negligible slip effects will exist at the solid-fluid interface. Accordingly, new theoretical models for thermoacoustic engines and heat pumps were derived, accounting for a slip boundary condition. These models are essential for the design process of micro-scale thermoacoustic devices that will operate under ultrasonic frequencies. Stability curves for engines - representing the onset of self-sustained oscillations - were calculated with both no-slip and slip boundary conditions, revealing improvement in the performance of engines with slip at the resonance frequency range applicable for micro-scale devices. Maximum achievable temperature differences curves for thermoacoustic heat pumps were calculated, revealing the negative effect of slip on the ability to pump heat up a temperature gradient. The authors acknowledge the support from the Nancy and Stephen Grand Technion Energy Program (GTEP).

  10. Fertilizer micro-dosing

    International Development Research Centre (IDRC) Digital Library (Canada)

    Localized application of small quantities of fertilizer (micro-dosing), combined with improved planting pits for rainwater harvesting, has generated greater profits and food security for women farmers in the Sahel. • Women are 25% more likely to use combined applications, and have expanded areas of food crops (cowpea,.

  11. Micro- and Nanoengineering

    NARCIS (Netherlands)

    Schroen, C.G.P.H.

    2015-01-01

    There are two overall themes, micro- and nanotechnology, which are capable of changing the future of food considerably. In microtechnology, production of foods and food ingredients is investigated at small scale; the results are thus that larger scale production is considered through operating many

  12. MicroRNA pharmacogenomics

    DEFF Research Database (Denmark)

    Rukov, Jakob Lewin; Shomron, Noam

    2011-01-01

    polymorphisms, copy number variations or differences in gene expression levels of drug metabolizing or transporting genes and drug targets. In this review paper, we focus instead on microRNAs (miRNAs): small noncoding RNAs, prevalent in metazoans, that negatively regulate gene expression in many cellular...

  13. Programming the BBC micro

    CERN Document Server

    Ferguson, John D; Macari, Louie; Williams, Peter H

    1983-01-01

    Programming the BBC Micro is a 12-chapter book that begins with a description of the BBC microcomputer, its peripheral, and faults. Subsequent chapters focus on practice in programming, program development, graphics, words, numbers, sound, bits, bytes, and assembly language. The interfacing, file handling, and detailed description of BBC microcomputer are also shown.

  14. Pyramid solar micro-grid

    Science.gov (United States)

    Huang, Bin-Juine; Hsu, Po-Chien; Wang, Yi-Hung; Tang, Tzu-Chiao; Wang, Jia-Wei; Dong, Xin-Hong; Hsu, Hsin-Yi; Li, Kang; Lee, Kung-Yen

    2018-03-01

    A novel pyramid solar micro-grid is proposed in the present study. All the members within the micro-grid can mutually share excess solar PV power each other through a binary-connection hierarchy. The test results of a 2+2 pyramid solar micro-grid consisting of 4 individual solar PV systems for self-consumption are reported.

  15. Micro Information Systems

    DEFF Research Database (Denmark)

    Ulslev Pedersen, Rasmus; Kühn Pedersen, Mogens

    2014-01-01

    such as medical and manufacturing. These new sensor applications have implications for information systems (IS) and, the authors visualize this new class of information systems as fractals growing from an established class of systems; namely that of information systems (IS). The identified applications...... and implications are used as an empirical basis for creating a model for these small new information systems. Such sensor systems are called embedded systems in the technical sciences, and the authors want to couple it with general IS. They call the merger of these two important research areas (IS and embedded...... systems) for micro information systems (micro-IS). It is intended as a new research field within IS research. An initial framework model is established, which seeks to capture both the possibilities and constraints of this new paradigm, while looking simultaneously at the fundamental IS and ICT aspects...

  16. CERN MicroClub

    CERN Multimedia

    CERN MicroClub

    2016-01-01

    Le CERN Micro Club (en partenariat avec Google Education et EU Code Week) organise un évènement éducatif exceptionnel autour de trois kits scientifiques basés sur le mini-ordinateur Raspberry Pi : Le Bras Robotique "Poppy Ergo Jr", conçu par l'équipe-projet Flowers (Centre de recherche Inria Bordeaux Sud-Ouest, ENSTA Paris Tech). Le kit de détection de rayons cosmiques "Muon Hunter", conçu en partenariat entre Mr Mihaly Vadai et les membres du CERN Micro Club. La voiture radio-commandée programmable Wifi "GianoPi", conçue en partenariat avec le campus "La Chataigneraie", pour l'Ecole Internationale de Genève.   Le vendredi 7 octobre (de 18h à 20h) : Une conférence gratuite et ouverte à tous (limitée à 100 personnes), pendant laquelle v...

  17. Micro dynamics in mediation

    OpenAIRE

    Boserup, Hans

    2014-01-01

    The author has identified a number of styles in mediation, which lead to different processes and different outcomes. Through discourse and conversation analysis he examines the micro dynamics in three of these, the postmodern styles: systemic, transformative and narrative mediation. The differences between the three mediation ideologies and practice is illustrated through role play scripts enacted in each style. Mediator and providers of mediation and trainers in mediation are encouraged to a...

  18. Rectenna session: Micro aspects

    Science.gov (United States)

    Gutmann, R. J.

    1980-01-01

    Two micro aspects of rectenna design are discussed: evaluation of the degradation in net rectenna RF to DC conversion efficiency due to power density variations across the rectenna (power combining analysis) and design of Yagi-Uda receiving elements to reduce rectenna cost by decreasing the number of conversion circuits (directional receiving elements). The first of these involves resolving a fundamental question of efficiency potential with a rectenna, while the second involves a design modification with a large potential cost saving.

  19. Flexible micro flow sensor for micro aerial vehicles

    Science.gov (United States)

    Zhu, Rong; Que, Ruiyi; Liu, Peng

    2017-12-01

    This article summarizes our studies on micro flow sensors fabricated on a flexible polyimide circuit board by a low-cost hybrid process of thin-film deposition and circuit printing. The micro flow sensor has merits of flexibility, structural simplicity, easy integrability with circuits, and good sensing performance. The sensor, which adheres to an object surface, can detect the surface flow around the object. In our study, we install the fabricated micro flow sensors on micro aerial vehicles (MAVs) to detect the surface flow variation around the aircraft wing and deduce the aerodynamic parameters of the MAVs in flight. Wind tunnel experiments using the sensors integrated with the MAVs are also conducted.

  20. Remote micro hydro

    Energy Technology Data Exchange (ETDEWEB)

    1985-03-01

    The micro-hydro project, built on a small tributary of Cowley Creek, near Whitehorse, Yukon, is an important step in the development of alternative energy sources and in conserving expensive diesel fuel. In addition to demonstrating the technical aspects of harnessing water power, the project paved the way for easier regulatory procedures. The power will be generated by a 9 meter head and a 6 inch crossflow turbine. The 36 V DC power will be stored in three 12 V batteries and converted to ac on demand by a 3,800 watt inverter. The system will produce 1.6 kW or 14,016 kWh per year with a firm flow of 1.26 cfs. This is sufficient to supply electricity for household needs and a wood working shop. The project is expected to cost about $18,000 and is more economical than tying into the present grid system, or continuing to use a gasoline generator. An environmental study determined that any impact of the project on the stream would be negligible. It is expected that no other water users will be affected by the project. This pilot project in micro-hydro applications will serve as a good indicator of the viability of this form of alternate energy in the Yukon. The calculations comparing the micro-hydro and grid system indicate that the mico-hydro system is a viable source of inflation-proof power. Higher heads and larger flow resulting in ac generation in excess of 10 kW would yield much better returns than this project. 3 tabs.

  1. micro strip gas chamber

    CERN Multimedia

    1998-01-01

    About 16 000 Micro Strip Gas Chambers like this one will be used in the CMS tracking detector. They will measure the tracks of charged particles to a hundredth of a millimetre precision in the region near the collision point where the density of particles is very high. Each chamber is filled with a gas mixture of argon and dimethyl ether. Charged particles passing through ionise the gas, knocking out electrons which are collected on the aluminium strips visible under the microscope. Such detectors are being used in radiography. They give higher resolution imaging and reduce the required dose of radiation.

  2. Review of Micro Magnetic Generator

    OpenAIRE

    Lin DU; Gengchen SHI; Jingjing ZHAO

    2014-01-01

    This paper discusses the research progress of micro magnetic generator systems. Micro magnetic generator systems convert energy from the environment to electric energy with advantages as high reliability, high power density, long life time and can be applied to extreme environment. This paper summarizes methods for improving generator performance of micro magnetic generator, including rotational magnetic generator, vibrational magnetic generator and hybrid magnetic generator, analyzes and com...

  3. Micro manufacturing techniques and applications

    CERN Document Server

    Du, Ruxu; Li, Zifu

    2013-01-01

    Micro/meso-scale manufacturing has been developed in research fields of machining, forming, materials and others, but its potential to industries are yet to be fully realized. The theme of the current volume was to build a bridge joining academic research and industrial needs in micro manufacturing. Among the 12 papers selected for publication are three keynote addresses onmicro and desktop factories for micro/meso-scale manufacturing applicationsand future visions, tissue cutting mechanics and applications for needlecore biopsy and guidance, and micro-texturing onto amorphous carbonmaterials

  4. Nozzle fabrication for Micro Propulsion of a Micro-Satellite

    NARCIS (Netherlands)

    Louwerse, M.C.; Jansen, Henricus V.; Groenendijk, M.N.W.; Elwenspoek, Michael Curt

    2008-01-01

    To enable formation flying of micro satellites, small sized propulsion systems are required. Our research focuses on the miniaturization of a feeding and thruster system by means of micro system technology (MST). Three fabrication methods have been investigated to make a conical converging-diverging

  5. A micro-coupling for micro mechanical systems

    Science.gov (United States)

    Li, Wei; Zhou, Zhixiong; Zhang, Bi; Xiao, Yunya

    2016-05-01

    The error motions of micro mechanical systems, such as micro-spindles, increase with the increasing of the rotational speed, which not only decreases the rotational accuracy, but also promotes instability and limits the maximum operational speed. One effective way to deal with it is to use micro-flexible couplings between the drive and driven shafts so as to reduce error motions of the driven shaft. But the conventional couplings, such as diaphragm couplings, elastomeric couplings, bellows couplings, and grooved couplings, etc, cannot be directly used because of their large and complicated structures. This study presents a novel micro-coupling that consists of a flexible coupling and a shape memory alloy (SMA)-based clamp for micro mechanical systems. It is monolithic and can be directly machined from a shaft. The study performs design optimization and provides manufacturing considerations, including thermo-mechanical training of the SMA ring for the desired Two-Way-Shape-Memory effect (TWSMe). A prototype micro-coupling and a prototype micro-spindle using the proposed coupling are fabricated and tested. The testing results show that the prototype micro-coupling can bear a torque of above 5 N • mm and an axial force of 8.5 N and be fitted with an SMA ring for clamping action at room temperature (15 °C) and unclamping action below-5 °C. At the same time, the prototype micro-coupling can work at a rotational speed of above 200 kr/min with the application to a high-speed precision micro-spindle. Moreover, the radial runout error of the artifact, as a substitute for the micro-tool, is less than 3 μm while that of turbine shaft is above 7 μm. It can be concluded that the micro-coupling successfully accommodates misalignment errors of the prototype micro-spindle. This research proposes a micro-coupling which is featured with an SMA ring, and it is designed to clamp two shafts, and has smooth transmission, simple assembly, compact structure, zero-maintenance and

  6. Applying a foil queue micro-electrode in micro-EDM to fabricate a 3D micro-structure

    Science.gov (United States)

    Xu, Bin; Guo, Kang; Wu, Xiao-yu; Lei, Jian-guo; Liang, Xiong; Guo, Deng-ji; Ma, Jiang; Cheng, Rong

    2018-05-01

    Applying a 3D micro-electrode in a micro electrical discharge machining (micro-EDM) can fabricate a 3D micro-structure with an up and down reciprocating method. However, this processing method has some shortcomings, such as a low success rate and a complex process for fabrication of 3D micro-electrodes. By focusing on these shortcomings, this paper proposed a novel 3D micro-EDM process based on the foil queue micro-electrode. Firstly, a 3D micro-electrode was discretized into several foil micro-electrodes and these foil micro-electrodes constituted a foil queue micro-electrode. Then, based on the planned process path, foil micro-electrodes were applied in micro-EDM sequentially and the micro-EDM results of each foil micro-electrode were able to superimpose the 3D micro-structure. However, the step effect will occur on the 3D micro-structure surface, which has an adverse effect on the 3D micro-structure. To tackle this problem, this paper proposes to reduce this adverse effect by rounded corner wear at the end of the foil micro-electrode and studies the impact of machining parameters on rounded corner wear and the step effect on the micro-structure surface. Finally, using a wire cutting voltage of 80 V, a current of 0.5 A and a pulse width modulation ratio of 1:4, the foil queue micro-electrode was fabricated by wire electrical discharge machining. Also, using a pulse width of 100 ns, a pulse interval of 200 ns, a voltage of 100 V and workpiece material of 304# stainless steel, the foil queue micro-electrode was applied in micro-EDM for processing of a 3D micro-structure with hemispherical features, which verified the feasibility of this process.

  7. New micro-organism

    Energy Technology Data Exchange (ETDEWEB)

    Takakuwa, Masayoshi; Hashimoto, Gotaro

    1987-09-12

    Invention relates with a new organism for the coal liquefying desulfurization. This micro-organism conducts a good sporulation on a culture medium which contains a coal as an only carbon source. It belongs to Penicillium and named Penicillium MT-6001 registered at Fermentation Research Institute No. 8463. Coal powder is thrown into a reaction vessel which accommodated a culture solution of this bacteria, and the surface of the solution is covered with liquid paraffin; coal powder is treated of liquefaction for about 5 hours while maintaining the anaerobic condition and slowly agitating to form a transparent solution layer on the surface of the reactor together with liquid paraffin. Liquefied product shows an analysis pattern similar to naphthenic petroleum containing a lipid with polar radical. (2 figs)

  8. MicroProteins

    DEFF Research Database (Denmark)

    Eguen, Teinai Ebimienere; Straub, Daniel; Graeff, Moritz

    2015-01-01

    MicroProteins (miPs) are short, usually single-domain proteins that, in analogy to miRNAs, heterodimerize with their targets and exert a dominant-negative effect. Recent bioinformatic attempts to identify miPs have resulted in a list of potential miPs, many of which lack the defining...... characteristics of a miP. In this opinion article, we clearly state the characteristics of a miP as evidenced by known proteins that fit the definition; we explain why modulatory proteins misrepresented as miPs do not qualify as true miPs. We also discuss the evolutionary history of miPs, and how the miP concept...

  9. Intronic microRNAs

    International Nuclear Information System (INIS)

    Ying, S.-Y.; Lin, S.-L.

    2005-01-01

    MicroRNAs (miRNAs), small single-stranded regulatory RNAs capable of interfering with intracellular mRNAs that contain partial complementarity, are useful for the design of new therapies against cancer polymorphism and viral mutation. MiRNA was originally discovered in the intergenic regions of the Caenorhabditis elegans genome as native RNA fragments that modulate a wide range of genetic regulatory pathways during animal development. However, neither RNA promoter nor polymerase responsible for miRNA biogenesis was determined. Recent findings of intron-derived miRNA in C. elegans, mouse, and human have inevitably led to an alternative pathway for miRNA biogenesis, which relies on the coupled interaction of Pol-II-mediated pre-mRNA transcription and intron excision, occurring in certain nuclear regions proximal to genomic perichromatin fibrils

  10. Micro thrust and heat generator

    Science.gov (United States)

    Garcia, E.J.

    1998-11-17

    A micro thrust and heat generator have a means for providing a combustion fuel source to an ignition chamber of the micro thrust and heat generator. The fuel is ignited by a ignition means within the micro thrust and heat generator`s ignition chamber where it burns and creates a pressure. A nozzle formed from the combustion chamber extends outward from the combustion chamber and tappers down to a narrow diameter and then opens into a wider diameter where the nozzle then terminates outside of said combustion chamber. The pressure created within the combustion chamber accelerates as it leaves the chamber through the nozzle resulting in pressure and heat escaping from the nozzle to the atmosphere outside the micro thrust and heat generator. The micro thrust and heat generator can be microfabricated from a variety of materials, e.g., of polysilicon, on one wafer using surface micromachining batch fabrication techniques or high aspect ratio micromachining techniques (LIGA). 30 figs.

  11. Sustainable Micro-Manufacturing of Micro-Components via Micro Electrical Discharge Machining

    Directory of Open Access Journals (Sweden)

    Valeria Marrocco

    2011-12-01

    Full Text Available Micro-manufacturing emerged in the last years as a new engineering area with the potential of increasing peoples’ quality of life through the production of innovative micro-devices to be used, for example, in the biomedical, micro-electronics or telecommunication sectors. The possibility to decrease the energy consumption makes the micro-manufacturing extremely appealing in terms of environmental protection. However, despite this common belief that the micro-scale implies a higher sustainability compared to traditional manufacturing processes, recent research shows that some factors can make micro-manufacturing processes not as sustainable as expected. In particular, the use of rare raw materials and the need of higher purity of processes, to preserve product quality and manufacturing equipment, can be a source for additional environmental burden and process costs. Consequently, research is needed to optimize micro-manufacturing processes in order to guarantee the minimum consumption of raw materials, consumables and energy. In this paper, the experimental results obtained by the micro-electrical discharge machining (micro-EDM of micro-channels made on Ni–Cr–Mo steel is reported. The aim of such investigation is to shed a light on the relation and dependence between the material removal process, identified in the evaluation of material removal rate (MRR and tool wear ratio (TWR, and some of the most important technological parameters (i.e., open voltage, discharge current, pulse width and frequency, in order to experimentally quantify the material waste produced and optimize the technological process in order to decrease it.

  12. Review of Micro Magnetic Generator

    Directory of Open Access Journals (Sweden)

    Lin DU

    2014-08-01

    Full Text Available This paper discusses the research progress of micro magnetic generator systems. Micro magnetic generator systems convert energy from the environment to electric energy with advantages as high reliability, high power density, long life time and can be applied to extreme environment. This paper summarizes methods for improving generator performance of micro magnetic generator, including rotational magnetic generator, vibrational magnetic generator and hybrid magnetic generator, analyzes and compares their design and performance, and concludes key technologies and ongoing challenges for further progress. The paper is instructive and meaningful to for research work of related field.

  13. Wear of micro end mills

    DEFF Research Database (Denmark)

    Bissacco, Giuliano; Hansen, Hans Nørgaard; De Chiffre, Leonardo

    2005-01-01

    This paper addresses the important issue of wear on micro end mills considering relevant metrological tools for its characterization and quantification. Investigation of wear on micro end mills is particularly difficult and no data are available in the literature. Small worn volumes cause large...... part. For this investigation 200 microns end mills are considered. Visual inspection of the micro tools requires high magnification and depth of focus. 3D reconstruction based on scanning electron microscope (SEM) images and stereo-pair technique is foreseen as a possible method for quantification...

  14. MicroRNA and cancer

    DEFF Research Database (Denmark)

    Jansson, Martin D; Lund, Anders H

    2012-01-01

    biological phenomena and pathologies. The best characterized non-coding RNA family consists in humans of about 1400 microRNAs for which abundant evidence have demonstrated fundamental importance in normal development, differentiation, growth control and in human diseases such as cancer. In this review, we...... summarize the current knowledge and concepts concerning the involvement of microRNAs in cancer, which have emerged from the study of cell culture and animal model systems, including the regulation of key cancer-related pathways, such as cell cycle control and the DNA damage response. Importantly, micro...

  15. Micro-mixer/combustor

    KAUST Repository

    Badra, Jihad Ahmad

    2014-09-18

    A micro-mixer/combustor to mix fuel and oxidant streams into combustible mixtures where flames resulting from combustion of the mixture can be sustained inside its combustion chamber is provided. The present design is particularly suitable for diffusion flames. In various aspects the present design mixes the fuel and oxidant streams prior to entering a combustion chamber. The combustion chamber is designed to prevent excess pressure to build up within the combustion chamber, which build up can cause instabilities in the flame. A restriction in the inlet to the combustion chamber from the mixing chamber forces the incoming streams to converge while introducing minor pressure drop. In one or more aspects, heat from combustion products exhausted from the combustion chamber may be used to provide heat to at least one of fuel passing through the fuel inlet channel, oxidant passing through the oxidant inlet channel, the mixing chamber, or the combustion chamber. In one or more aspects, an ignition strip may be positioned in the combustion chamber to sustain a flame without preheating.

  16. Micro transport phenomena during boiling

    CERN Document Server

    Peng, Xiaofeng

    2011-01-01

    "Micro Transport Phenomena During Boiling" reviews the new achievements and contributions in recent investigations at microscale. It presents some original research results and discusses topics at the frontier of thermal and fluid sciences.

  17. Micro Mercury Ion Clock (MMIC)

    Data.gov (United States)

    National Aeronautics and Space Administration — Demonstrate micro clock based on trapped Hg ions with more than 10x size reduction and power; Fractional frequency stability at parts per 1014 level, adequate for...

  18. Micro-gen metering solutions

    Energy Technology Data Exchange (ETDEWEB)

    Elland, J.; Dickson, J.; Cranfield, P.

    2003-07-01

    This report summarises the results of a project to investigate the regulation of domestic electricity metering work and identify the most economic options for micro-generator installers to undertake work on electricity meters. A micro-generation unit is defined as an energy conversion system converting non-electrical energy into electrical energy and can include technologies such as photovoltaic systems, small-scale wind turbines, micro-hydroelectric systems, and combined heat and power systems. Details of six tasks are given and cover examination of the existing framework and legal documentation for metering work, the existing technical requirements for meter operators, meter operator personnel accreditation, appraisal of options for meter changes and for micro-generation installation, document change procedures, industry consultation, and a review of the costs implications of the options.

  19. Micro creep mechanisms of tungsten

    International Nuclear Information System (INIS)

    Levoy, R.; Hugon, I.; Burlet, H.; Baillin, X.; Guetaz, L.

    2000-01-01

    Due to its high melting point (3410 deg C), tungsten offers good mechanical properties at elevated temperatures for several applications in non-oxidizing environment. The creep behavior of tungsten is well known between 1200 and 2500 deg C and 10 -3 to 10 -1 strain. However, in some applications when dimensional stability of components is required, these strains are excessive and it is necessary to know the creep behavior of the material for micro-strains (between 10 -4 and 10 -6 ). Methods and devices used to measure creep micro-strains are presented, and creep equations (Norton and Chaboche laws) were developed for wrought, annealed and recrystallized tungsten. The main results obtained on tungsten under low stresses are: stress exponent 1, symmetry of micro-strains in creep-tension and creep-compression, inverse creep (threshold stress), etc. TEM, SEM and EBSD studies allow interpretation of the micro-creep mechanism of tungsten under low stresses and low temperature (∼0.3 K) like the Harper-Dorn creep. In Harper-Dorn creep, micro-strains are associated with the density and the distribution of dislocations existing in the crystals before creep. At 975 deg C, the initial dislocation structure moves differently whether or not a stress is applied. To improve the micro-creep behavior of tungsten, a heat treatment is proposed to create the optimum dislocation structure. (authors)

  20. Micro watt thermocurrent generator

    International Nuclear Information System (INIS)

    Bustard, T.; Goslee, D.; Barr, H.

    1976-01-01

    This nuclear thermocurrent generator to feed a cardic pacemaker should have higher life expectancy and reliability than was previously achieved. For this purpose a gettering arrangement is connected to be heat conducting immediately adjacent to the nuclear fuel arrangement in an evacuated casing. The gettering arrangement can be operated to activate at as high a temperature as possible, from 121 0 C to preferably about 204 0 C, so that a high vacuum is maintained. The current generating thermal column works at a temperature difference of 55.6 0 C. As the cold end of the column is connected to the outer casing, and should be held to a mean body temperature of 37.8 0 C, the hot side of the thermal column may only be heated to 93.4 0 C. The temperature jump from 121 0 or 204 0 to 93.4 0 is produced by a thermal resistance inserted between the hot side of the thermal column and the fuel arrangement. It may consist of a spacer made of stainless steel or by a gap, while in this first arrangement the nuclear heat generator is situated between the gettering arrangement and the thermal column, another arrangement shows the gettering arrangement enclosed in the fuel arrangement and thermal column. Here the heat flows in one direction only, the required temperature gradient is produced by suitable construction of the heat contacts between the 3 elements. Detailed constructional and manufacturing data are given for both models. Plutonium oxide is welded into a double casing as heat generator, in example the casing is made of nickel alloy. 1/10 gram of plutonium supplies a thermal energy of 50m watts, which produces a thermal current of 300 to 400 micro watts at 0.3V. (RW) [de

  1. Micro-droplet formation via 3D printed micro channel

    Science.gov (United States)

    Jian, Zhen; Zhang, Jiaming; Li, Erqiang; Thoroddsen, Sigurdur T.

    2016-11-01

    Low cost, fast-designed and fast-fabricated 3D micro channel was used to create micro-droplets. Capillary with an outer diameter of 1.5 mm and an inner diameter of 150 μm was inserted into a 3D printed cylindrical channel with a diameter of 2 mm . Flow rate of the two inlets, insert depth, liquid (density, viscosity and surface tension) and solid (roughness, contact angle) properties all play a role in the droplet formation. Different regimes - dripping, jetting, unstable state - were observed in the micro-channel on varying these parameters. With certain parameter combinations, successive formation of micro-droplets with equal size was observed and its size can be much smaller than the smallest channel size. Based on our experimental results, the droplet formation via 3D printed micro T-junction was investigated through direct numerical simulations with a code called Gerris. Reynolds numbers Re = ρUL / μ and Weber numbers We = ρU2 L / σ of the two liquids were introduced to measure the liquid effect. The parameter regime where different physical dynamics occur was studied and the regime transition was observed with certain threshold values. Qualitative and quantitative analysis were performed as well between simulations and experiments.

  2. Micro Machining Enhances Precision Fabrication

    Science.gov (United States)

    2007-01-01

    Advanced thermal systems developed for the Space Station Freedom project are now in use on the International Space Station. These thermal systems employ evaporative ammonia as their coolant, and though they employ the same series of chemical reactions as terrestrial refrigerators, the space-bound coolers are significantly smaller. Two Small Business Innovation Research (SBIR) contracts between Creare Inc. of Hanover, NH and Johnson Space Center developed an ammonia evaporator for thermal management systems aboard Freedom. The principal investigator for Creare Inc., formed Mikros Technologies Inc. to commercialize the work. Mikros Technologies then developed an advanced form of micro-electrical discharge machining (micro-EDM) to make tiny holes in the ammonia evaporator. Mikros Technologies has had great success applying this method to the fabrication of micro-nozzle array systems for industrial ink jet printing systems. The company is currently the world leader in fabrication of stainless steel micro-nozzles for this market, and in 2001 the company was awarded two SBIR research contracts from Goddard Space Flight Center to advance micro-fabrication and high-performance thermal management technologies.

  3. Cavitational micro-particles: plasma formation mechanisms

    International Nuclear Information System (INIS)

    Bica, Ioan

    2005-01-01

    Cavitational micro-particles are a class to which the micro-spheres, the micro-tubes and the octopus-shaped micro-particles belong. The cavitational micro-particles (micro-spheres, micro-tubes and octopus-shaped micro-particles) at an environmental pressure. The micro-spheres, the micro-tubes and the ligaments of the octopus-shaped micro-particles are produced in the argon plasma and are formed of vapors with low values of the molar concentration in comparison with the molar density of the gas and vapor mixture, the first one on the unstable and the last two on the stable movement of the vapors. The ligaments of the octopus-shaped micro-particles are open at the top for well-chosen values of the sub-cooling of the vapor and gas cylinders. The nitrogen in the air favors the formation of pores in the wall of the micro-spheres. In this paper we present the cavitational micro-particles, their production in the plasma and some mechanisms for their formation in the plasma. (author)

  4. MicroPRIS user's guide

    International Nuclear Information System (INIS)

    1991-01-01

    MicroPRIS is a new service of the IAEA Power Reactor Information System (PRIS) for the Member States of IAEA. MicroPRIS makes the IAEA database on nuclear power plants and their operating experience available to Member States on computer diskettes in a form readily accessible by standard commercially available personal computer packages. The aim of this publication is to provide the users of the PC version of PRIS data with description of the subset of the full PRIS database contained in MicroPRIS (release 1990), description of files and file structures, field descriptions and definitions, extraction and selection guide and with the method of calculation of a number of important performance indicators used by the IAEA

  5. Dimensional micro and nano metrology

    DEFF Research Database (Denmark)

    Hansen, Hans Nørgaard; da Costa Carneiro, Kim; Haitjema, Han

    2006-01-01

    The need for dimensional micro and nano metrology is evident, and as critical dimensions are scaled down and geometrical complexity of objects is increased, the available technologies appear not sufficient. Major research and development efforts have to be undertaken in order to answer these chal......The need for dimensional micro and nano metrology is evident, and as critical dimensions are scaled down and geometrical complexity of objects is increased, the available technologies appear not sufficient. Major research and development efforts have to be undertaken in order to answer...... these challenges. The developments have to include new measuring principles and instrumentation, tolerancing rules and procedures as well as traceability and calibration. The current paper describes issues and challenges in dimensional micro and nano metrology by reviewing typical measurement tasks and available...

  6. Micro-Mechanical Temperature Sensors

    DEFF Research Database (Denmark)

    Larsen, Tom

    Temperature is the most frequently measured physical quantity in the world. The field of thermometry is therefore constantly evolving towards better temperature sensors and better temperature measurements. The aim of this Ph.D. project was to improve an existing type of micro-mechanical temperature...... sensor or to develop a new one. Two types of micro-mechanical temperature sensors have been studied: Bilayer cantilevers and string-like beam resonators. Both sensor types utilize thermally generated stress. Bilayer cantilevers are frequently used as temperature sensors at the micro-scale, and the goal....... The reduced sensitivity was due to initial bending of the cantilevers and poor adhesion between the two cantilever materials. No further attempts were made to improve the sensitivity of bilayer cantilevers. The concept of using string-like resonators as temperature sensors has, for the first time, been...

  7. Micro-machined calorimetric biosensors

    Science.gov (United States)

    Doktycz, Mitchel J.; Britton, Jr., Charles L.; Smith, Stephen F.; Oden, Patrick I.; Bryan, William L.; Moore, James A.; Thundat, Thomas G.; Warmack, Robert J.

    2002-01-01

    A method and apparatus are provided for detecting and monitoring micro-volumetric enthalpic changes caused by molecular reactions. Micro-machining techniques are used to create very small thermally isolated masses incorporating temperature-sensitive circuitry. The thermally isolated masses are provided with a molecular layer or coating, and the temperature-sensitive circuitry provides an indication when the molecules of the coating are involved in an enthalpic reaction. The thermally isolated masses may be provided singly or in arrays and, in the latter case, the molecular coatings may differ to provide qualitative and/or quantitative assays of a substance.

  8. Automated Micro Hall Effect measurements

    DEFF Research Database (Denmark)

    Petersen, Dirch Hjorth; Henrichsen, Henrik Hartmann; Lin, Rong

    2014-01-01

    With increasing complexity of processes and variety of materials used for semiconductor devices, stringent control of the electronic properties is becoming ever more relevant. Collinear micro four-point probe (M4PP) based measurement systems have become high-end metrology methods for characteriza......With increasing complexity of processes and variety of materials used for semiconductor devices, stringent control of the electronic properties is becoming ever more relevant. Collinear micro four-point probe (M4PP) based measurement systems have become high-end metrology methods...

  9. Improvement of micro endmill geometry for micro hard milling application

    NARCIS (Netherlands)

    Li, P.; Oosterling, J.A.J.; Hoogstrate, A.M.; Langen, H.H.

    2008-01-01

    One of the applications of the micromilling technology is to machine micro features on moulds by direct machining of hardened tool steels. However at this moment, this process is not industrial applicable because of the encountered problems, such as the big tool deflection, severe tool wear, and

  10. On-chip micro-power: three-dimensional structures for micro-batteries and micro-supercapacitors

    Science.gov (United States)

    Beidaghi, Majid; Wang, Chunlei

    2010-04-01

    With the miniaturization of portable electronic devices, there is a demand for micro-power source which can be integrated on the semiconductor chips. Various micro-batteries have been developed in recent years to generate or store the energy that is needed by microsystems. Micro-supercapacitors are also developed recently to couple with microbatteries and energy harvesting microsystems and provide the peak power. Increasing the capacity per footprint area of micro-batteries and micro-supercapacitors is a great challenge. One promising route is the manufacturing of three dimensional (3D) structures for these micro-devices. In this paper, the recent advances in fabrication of 3D structure for micro-batteries and micro-supercapacitors are briefly reviewed.

  11. Micro-machined resonator oscillator

    Science.gov (United States)

    Koehler, Dale R.; Sniegowski, Jeffry J.; Bivens, Hugh M.; Wessendorf, Kurt O.

    1994-01-01

    A micro-miniature resonator-oscillator is disclosed. Due to the miniaturization of the resonator-oscillator, oscillation frequencies of one MHz and higher are utilized. A thickness-mode quartz resonator housed in a micro-machined silicon package and operated as a "telemetered sensor beacon" that is, a digital, self-powered, remote, parameter measuring-transmitter in the FM-band. The resonator design uses trapped energy principles and temperature dependence methodology through crystal orientation control, with operation in the 20-100 MHz range. High volume batch-processing manufacturing is utilized, with package and resonator assembly at the wafer level. Unique design features include squeeze-film damping for robust vibration and shock performance, capacitive coupling through micro-machined diaphragms allowing resonator excitation at the package exterior, circuit integration and extremely small (0.1 in. square) dimensioning. A family of micro-miniature sensor beacons is also disclosed with widespread applications as bio-medical sensors, vehicle status monitors and high-volume animal identification and health sensors. The sensor family allows measurement of temperatures, chemicals, acceleration and pressure. A microphone and clock realization is also available.

  12. Hybrid cycles for micro generation

    International Nuclear Information System (INIS)

    Campanari, S.

    2000-01-01

    This paper deals with the main features of two emerging technologies in the field of small-scale power generation, micro turbines and Solid Oxide Fuel Cells, discussing the extremely high potential of their combination into hybrid cycles and their possible role for distributed cogeneration [it

  13. Micro-Encapsulation of Probiotics

    Science.gov (United States)

    Meiners, Jean-Antoine

    Micro-encapsulation is defined as the technology for packaging with the help of protective membranes particles of finely ground solids, droplets of liquids or gaseous materials in small capsules that release their contents at controlled rates over prolonged periods of time under the influences of specific conditions (Boh, 2007). The material encapsulating the core is referred to as coating or shell.

  14. Micro RNAs in animal development.

    NARCIS (Netherlands)

    Plasterk, R.H.A.

    2006-01-01

    Micro RNAs (miRNAs) are approximately 22 nucleotide single-stranded noncoding RNA molecules that bind to target messenger RNAs (mRNAs) and silence their expression. This Essay explores the importance of miRNAs in animal development and their possible roles in disease and evolution.

  15. Micro Coriolis Gas Density Sensor

    NARCIS (Netherlands)

    Sparreboom, Wouter; Ratering, Gijs; Kruijswijk, Wim; van der Wouden, E.J.; Groenesteijn, Jarno; Lötters, Joost Conrad

    2017-01-01

    In this paper we report on gas density measurements using a micro Coriolis sensor. The technology to fabricate the sensor is based on surface channel technology. The measurement tube is freely suspended and has a wall thickness of only 1 micron. This renders the sensor extremely sensitive to changes

  16. Contamination Study of Micro Pulsed Plasma Thruster

    National Research Council Canada - National Science Library

    Kesenek, Ceylan

    2008-01-01

    .... Micro-Pulsed Plasma Thrusters (PPTs) are highly reliable and simple micro propulsion systems that will offer attitude control, station keeping, constellation flying, and drag compensation for such satellites...

  17. Micro-Rockets for the Classroom.

    Science.gov (United States)

    Huebner, Jay S.; Fletcher, Alice S.; Cato, Julia A.; Barrett, Jennifer A.

    1999-01-01

    Compares micro-rockets to commercial models and water rockets. Finds that micro-rockets are more advantageous because they are constructed with inexpensive and readily available materials and can be safely launched indoors. (CCM)

  18. Equivalent Simplification Method of Micro-Grid

    OpenAIRE

    Cai Changchun; Cao Xiangqin

    2013-01-01

    The paper concentrates on the equivalent simplification method for the micro-grid system connection into distributed network. The equivalent simplification method proposed for interaction study between micro-grid and distributed network. Micro-grid network, composite load, gas turbine synchronous generation, wind generation are equivalent simplification and parallel connect into the point of common coupling. A micro-grid system is built and three phase and single phase grounded faults are per...

  19. Development of micro pattern cutting simulation software

    International Nuclear Information System (INIS)

    Lee, Jong Min; Song, Seok Gyun; Choi, Jeong Ju; Novandy, Bondhan; Kim, Su Jin; Lee, Dong Yoon; Nam, Sung Ho; Je, Tae Jin

    2008-01-01

    The micro pattern machining on the surface of wide mold is not easy to be simulated by conventional software. In this paper, a software is developed for micro pattern cutting simulation. The 3d geometry of v-groove, rectangular groove, pyramid and pillar patterns are visualized by c++ and OpenGL library. The micro cutting force is also simulated for each pattern

  20. The MicroObservatory Net

    Science.gov (United States)

    Brecher, K.; Sadler, P.

    1994-12-01

    A group of scientists, engineers and educators based at the Harvard-Smithsonian Center for Astrophysics (CfA) has developed a prototype of a small, inexpensive and fully integrated automated astronomical telescope and image processing system. The project team is now building five second generation instruments. The MicroObservatory has been designed to be used for classroom instruction by teachers as well as for original scientific research projects by students. Probably in no other area of frontier science is it possible for a broad spectrum of students (not just the gifted) to have access to state-of-the-art technologies that would allow for original research. The MicroObservatory combines the imaging power of a cooled CCD, with a self contained and weatherized reflecting optical telescope and mount. A microcomputer points the telescope and processes the captured images. The MicroObservatory has also been designed to be used as a valuable new capture and display device for real time astronomical imaging in planetariums and science museums. When the new instruments are completed in the next few months, they will be tried with high school students and teachers, as well as with museum groups. We are now planning to make the MicroObservatories available to students, teachers and other individual users over the Internet. We plan to allow the telescope to be controlled in real time or in batch mode, from a Macintosh or PC compatible computer. In the real-time mode, we hope to give individual access to all of the telescope control functions without the need for an "on-site" operator. Users would sign up for a specific period of time. In the batch mode, users would submit jobs for the telescope. After the MicroObservatory completed a specific job, the images would be e-mailed back to the user. At present, we are interested in gaining answers to the following questions: (1) What are the best approaches to scheduling real-time observations? (2) What criteria should be used

  1. Isolation of microRNA targets using biotinylated synthetic microRNAs

    DEFF Research Database (Denmark)

    Ørom, Ulf Andersson; Lund, Anders H

    2007-01-01

    MicroRNAs are small regulatory RNAs found in multicellular organisms where they post-transcriptionally regulate gene expression. In animals, microRNAs bind mRNAs via incomplete base pairings making the identification of microRNA targets inherently difficult. Here, we present a detailed method...... for experimental identification of microRNA targets based on affinity purification of tagged microRNAs associated with their targets. Udgivelsesdato: 2007-Oct...

  2. Integration of micro milling highspeed spindle on a microEDM-milling machine set-up

    DEFF Research Database (Denmark)

    De Grave, Arnaud; Hansen, Hans Nørgaard; Andolfatto, Loic

    2009-01-01

    In order to cope with repositioning errors and to combine the fast removal rate of micro milling with the precision and small feature size achievable with micro EDM milling, a hybrid micro-milling and micro-EDM milling centre was built and tested. The aim was to build an affordable set-up, easy...... by micro milling. Examples of test parts are shown and used as an experimental validation....

  3. Voltammetry at micro-mesh electrodes

    Directory of Open Access Journals (Sweden)

    Wadhawan Jay D.

    2003-01-01

    Full Text Available The voltammetry at three micro-mesh electrodes is explored. It is found that at sufficiently short experimental durations, the micro-mesh working electrode first behaves as an ensemble of microband electrodes, then follows the behaviour anticipated for an array of diffusion-independent micro-ring electrodes of the same perimeter as individual grid-squares within the mesh. During prolonged electrolysis, the micro-mesh electrode follows that behaviour anticipated theoretically for a cubically-packed partially-blocked electrode. Application of the micro-mesh electrode for the electrochemical determination of carbon dioxide in DMSO electrolyte solutions is further illustrated.

  4. Micro processors for plant protection

    International Nuclear Information System (INIS)

    McAffer, N.T.C.

    1976-01-01

    Micro computers can be used satisfactorily in general protection duties with economic advantages over hardwired systems. The reliability of such protection functions can be enhanced by keeping the task performed by each protection micro processor simple and by avoiding such a task being dependent on others in any substantial way. This implies that vital work done for any task is kept within it and that any communications from it to outside or to it from outside are restricted to those for controlling data transfer. Also that the amount of this data should be the minimum consistent with satisfactory task execution. Technology is changing rapidly and devices may become obsolete and be supplanted by new ones before their theoretical reliability can be confirmed or otherwise by field service. This emphasises the need for users to pool device performance data so that effective reliability judgements can be made within the lifetime of the devices. (orig.) [de

  5. Epigenetic microRNA Regulation

    DEFF Research Database (Denmark)

    Wiklund, Erik Digman

    2011-01-01

    MicroRNAs (miRNAs) are small non-coding RNAs (ncRNAs) that negatively regulate gene expression post-transcriptionally by binding to complementary sequences in the 3’UTR of target mRNAs in the cytoplasm. However, recent evidence suggests that certain miRNAs are enriched in the nucleus, and their t......MicroRNAs (miRNAs) are small non-coding RNAs (ncRNAs) that negatively regulate gene expression post-transcriptionally by binding to complementary sequences in the 3’UTR of target mRNAs in the cytoplasm. However, recent evidence suggests that certain miRNAs are enriched in the nucleus...

  6. Combinatorial microRNA target predictions

    DEFF Research Database (Denmark)

    Krek, Azra; Grün, Dominic; Poy, Matthew N.

    2005-01-01

    MicroRNAs are small noncoding RNAs that recognize and bind to partially complementary sites in the 3' untranslated regions of target genes in animals and, by unknown mechanisms, regulate protein production of the target transcript1, 2, 3. Different combinations of microRNAs are expressed...... in different cell types and may coordinately regulate cell-specific target genes. Here, we present PicTar, a computational method for identifying common targets of microRNAs. Statistical tests using genome-wide alignments of eight vertebrate genomes, PicTar's ability to specifically recover published micro......RNA targets, and experimental validation of seven predicted targets suggest that PicTar has an excellent success rate in predicting targets for single microRNAs and for combinations of microRNAs. We find that vertebrate microRNAs target, on average, roughly 200 transcripts each. Furthermore, our results...

  7. Micro Grid: A Smart Technology

    OpenAIRE

    Naveenkumar, M; Ratnakar, N

    2012-01-01

    Distributed Generation (DG) is an approach that employs small-scale technologies to produce electricity close to the end users of power. Todays DG technologies often consist of renewable generators, and offer a number of potential benefits. This paper presents a design of micro grid as part of Smart grid technologies with renewable energy resources like solar, wind and Diesel generator. The design of the microgrid with integration of Renewable energy sources are done in PSCAD/EMTDC.This paper...

  8. Introduction to micro- and nanooptics

    CERN Document Server

    Jahns, Jürgen

    2012-01-01

    This first textbook on both micro- and nanooptics introduces readers to the technological development, physical background and key areas.The opening chapters on the physics of light are complemented by chapters on refractive and diffractive optical elements. The internationally renowned authors present different methods of lithographic and nonlithographic fabrication of microoptics and introduce the characterization and testing of microoptics.The second part of the book is dedicated to optical microsystems and MEMS, optical waveguide structures and optical nanostructures, including pho

  9. The useful micro-organism

    International Nuclear Information System (INIS)

    1970-01-01

    Can man survive civilization? Academician Ivan Malek, Director of the Institute of Microbiology in Prague, a member of the Agency's Scientific Advisory Committee and for many years an adviser to the Food and Agriculture Organization, the World Health Organization and UNESCO, believes he can, But he also considers that if man is to survive he must study and use all the resources at his disposal - including the micro-organisms of the planet earth. (author)

  10. The Micro Trench Gas Counter

    International Nuclear Information System (INIS)

    Schmitz, J.

    1991-07-01

    A novel design is presented for a gas avalanche chamber with micro-strip gas readout. While existing gaseous microstrip detectors (Micro-strip Gas Counters, Knife edge chambers) have a minimum anode pitch of the order of 100 μm, the pitch of the discussed Micro Trench Gas Counter goes down to 30-50 μm. This leads to a better position resolution and two track separation, and a higher radiation resistivity. Its efficiency and signal speed are expected to be the same as the Microstrip Gas Counter. The energy resolution of the device is expected to be equal to or better than 10 percent for the 55 Fe peak. Since the anode strip dimensions are larger than those in a MSGC, the device may be not as sensitive to discharges and mechanical damage. In this report production of the device is briefly described, and predictions on its operation are made based on electric field calculations and experience with the Microstrip Gas Counter. The authors restrict themselves to the application in High Energy Physics. (author). 10 refs.; 9 figs

  11. Application of flexible micro temperature sensor in oxidative steam reforming by a methanol micro reformer.

    Science.gov (United States)

    Lee, Chi-Yuan; Lee, Shuo-Jen; Shen, Chia-Chieh; Yeh, Chuin-Tih; Chang, Chi-Chung; Lo, Yi-Man

    2011-01-01

    Advances in fuel cell applications reflect the ability of reformers to produce hydrogen. This work presents a flexible micro temperature sensor that is fabricated based on micro-electro-mechanical systems (MEMS) technology and integrated into a flat micro methanol reformer to observe the conditions inside that reformer. The micro temperature sensor has higher accuracy and sensitivity than a conventionally adopted thermocouple. Despite various micro temperature sensor applications, integrated micro reformers are still relatively new. This work proposes a novel method for integrating micro methanol reformers and micro temperature sensors, subsequently increasing the methanol conversion rate and the hydrogen production rate by varying the fuel supply rate and the water/methanol ratio. Importantly, the proposed micro temperature sensor adequately controls the interior temperature during oxidative steam reforming of methanol (OSRM), with the relevant parameters optimized as well.

  12. Application of Flexible Micro Temperature Sensor in Oxidative Steam Reforming by a Methanol Micro Reformer

    Directory of Open Access Journals (Sweden)

    Yi-Man Lo

    2011-02-01

    Full Text Available Advances in fuel cell applications reflect the ability of reformers to produce hydrogen. This work presents a flexible micro temperature sensor that is fabricated based on micro-electro-mechanical systems (MEMS technology and integrated into a flat micro methanol reformer to observe the conditions inside that reformer. The micro temperature sensor has higher accuracy and sensitivity than a conventionally adopted thermocouple. Despite various micro temperature sensor applications, integrated micro reformers are still relatively new. This work proposes a novel method for integrating micro methanol reformers and micro temperature sensors, subsequently increasing the methanol conversion rate and the hydrogen production rate by varying the fuel supply rate and the water/methanol ratio. Importantly, the proposed micro temperature sensor adequately controls the interior temperature during oxidative steam reforming of methanol (OSRM, with the relevant parameters optimized as well.

  13. Micro tooling technologies for polymer micro replication: direct, indirect and hybrid process chains

    DEFF Research Database (Denmark)

    Tosello, Guido; Hansen, Hans Nørgaard

    2009-01-01

    The increasing employment of micro products, of products containing micro parts and of products with micro-structured surfaces calls for mass fabrication technologies based on replication processes. In many cases, a suitable solution is given by the use of polymer micro products, whose production...... and performance of the corresponding micro mould. Traditional methods of micro tooling, such as various machining processes (e.g. micro milling, micro electrical discharge machining) have already reached their limitations with decreasing dimensions of mould inserts and cavities. To this respect, tooling process...... chains based on combination of micro manufacturing processes (defined as hybrid tooling) have been established in order to obtain further features miniaturization and increased accuracy. In this paper, examples and performance of different hybrid tooling approaches as well as challenges, opportunities...

  14. Thermal performance of a micro-combustor for micro-gas turbine system

    International Nuclear Information System (INIS)

    Cao, H.L.; Xu, J.L.

    2007-01-01

    Premixed combustion of hydrogen gas and air was performed in a stainless steel based micro-annular combustor for a micro-gas turbine system. Micro-scale combustion has proved to be stable in the micro-combustor with a gap of 2 mm. The operating range of the micro-combustor was measured, and the maximum excess air ratio is up to 4.5. The distribution of the outer wall temperature and the temperature of exhaust gas of the micro-combustor with excess air ratio were obtained, and the wall temperature of the micro-combustor reaches its maximum value at the excess air ratio of 0.9 instead of 1 (stoichiometric ratio). The heat loss of the micro-combustor to the environment was calculated and even exceeds 70% of the total thermal power computed from the consumed hydrogen mass flow rate. Moreover, radiant heat transfer covers a large fraction of the total heat loss. Measures used to reduce the heat loss were proposed to improve the thermal performance of the micro-combustor. The optimal operating status of the micro-combustor and micro-gas turbine is analyzed and proposed by analyzing the relationship of the temperature of the exhaust gas of the micro-combustor with thermal power and excess air ratio. The investigation of the thermal performance of the micro-combustor is helpful to design an improved micro-combustor

  15. Micro-PNT and Comprehensive PNT

    Directory of Open Access Journals (Sweden)

    YANG Yuanxi

    2017-10-01

    Full Text Available Comprehensive or integrated positioning, navigation and timing is an obvious developing trend following the global navigation satellite system.This paper summarizes the current status of micro-PNT and its developing requirements. The related key technologies are described and the relationship between comprehensive PNT and micro-PNT is analyzed. It is stressed that the comprehensive PNT needs massive infrastructure construction and investment, however, the micro-PNT aims at the integrated applications of high-tech micro sensors. It is different from the current opinions appeared in the literatures, micro-PNT should include multi GNSS integration and micro components of navigation and timing in order to make the PNT outputs refer to a unified coordinate datum and time scale. Micro-PNT focuses on the personalized micro terminal applications. Except for the miniaturization of each PNT component, micro-PNT aims at the deep integration of the micro sensors, adaptive data fusion and self calibration of each component.

  16. Bottoming micro-Rankine cycles for micro-gas turbines

    International Nuclear Information System (INIS)

    Invernizzi, Costante; Iora, Paolo; Silva, Paolo

    2007-01-01

    This paper investigates the possibility of enhancing the performances of micro-gas turbines through the addition of a bottoming organic Rankine cycle which recovers the thermal power of the exhaust gases typically available in the range of 250-300 o C. The ORC cycles are particularly suitable for the recovery of heat from sources at variable temperatures, and for the generation of medium to small electric power. With reference to a micro-gas turbine with a size of about 100 kWe, a combined configuration could increase the net electric power by about 1/3, yielding an increase of the electrical efficiency of up to 40%. A specific analysis of the characteristics of different classes of working fluids is carried out in order to define a procedure to select the most appropriate fluid, capable of satisfying both environmental (ozone depletion potential, global warming potential) and technical (flammability, toxicity, fluid critical temperature and molecular complexity) concerns. Afterwards, a thermodynamic analysis is performed to ascertain the most favourable cycle thermodynamic conditions, from the point of view of heat recovery. Furthermore, a preliminary design of the ORC turbine (number of stages, outer diameter and rotational speed) is carried out

  17. Non-Photolithographic Manufacturing Processes for Micro-Channels Functioned by Micro-Contact-Printed SAMs

    Science.gov (United States)

    Saigusa, Hiroki; Suga, Yasuo; Miki, Norihisa

    In this paper we propose non-photolithographic fabrication processes of micro-fluid channels with patterned SAMs (Self-Assembled-Monolayers). SAMs with a thiol group are micro-contact printed on a patterned Au/Ti layer, which is vapor-deposited through a shadow mask. Ti is an adhesion layer. Subsequently, the micro-channels are formed by bonding surface-activated PDMS onto the silicon substrate via a silanol group, producing a SAMs-functioned bottom wall of the micro-channel. No photolithographic processes are necessary and thus, the proposed processes are very simple, quick and low cost. The micro-reactors can have various functions associated with the micro-contact-printed SAMs. We demonstrate successful manufacturing of micro-reactors with two types of SAMs. The micro-reactor with patterned AUT (11-amino-1-undecanethiol) successfully trapped nano-particles with a carboxylic acid group, indicating that micro-contact-printed SAMs remain active after the manufacturing processes of the micro-reactor. AUT -functioned micro-channels are applicable to bioassay and to immobilize proteins for DNA arrays. ODT (1-octadecanethiol) makes surfaces hydrophobic with the methyl terminal group. When water was introduced into the micro-reactor with ODT-patterned surfaces, water droplets remained only in the hydrophilic areas where ODT was not patterned. ODT -functioned micro-channels are applicable to fluid handling.

  18. Macro-Micro Interlocked Simulator

    International Nuclear Information System (INIS)

    Sato, Tetsuya

    2005-01-01

    Simulation Science is now standing on a turning point. After the appearance of the Earth Simulator, HEC is struggling with several severe difficulties due to the physical limit of LSI technologies and the so-called latency problem. In this paper I would like to propose one clever way to overcome these difficulties from the simulation algorithm viewpoint. Nature and artificial products are usually organized with several nearly autonomously working internal systems (organizations, or layers). The Earth Simulator has gifted us with a really useful scientific tool that can deal with the entire evolution of one internal system with a sufficient soundness. In order to make a leap jump of Simulation Science, therefore, it is desired to design an innovative simulator that enables us to deal with simultaneously and as consistently as possible a real system that evolves cooperatively with several internal autonomous systems. Three years experience of the Earth Simulator Project has stimulated to come up with one innovative simulation algorithm to get rid of the technological barrier standing in front of us, which I would like to call 'Macro-Micro Interlocked Algorithm', or 'Macro-Micro Multiplying Algorithm', and present a couple of such examples to validate the proposed algorithm. The first example is an aurora-arc formation as a result of the mutual interaction between the macroscopic magnetosphere-ionosphere system and the microscopic field-aligned electron and ion system. The second example is the local heavy rain fall resulting from the interaction between the global climate evolution and the microscopic raindrop growth process. Based on this innovative feasible algorithm, I came up with a Macro-Micro Multiplying Simulator

  19. Wafer integrated micro-scale concentrating photovoltaics

    Science.gov (United States)

    Gu, Tian; Li, Duanhui; Li, Lan; Jared, Bradley; Keeler, Gordon; Miller, Bill; Sweatt, William; Paap, Scott; Saavedra, Michael; Das, Ujjwal; Hegedus, Steve; Tauke-Pedretti, Anna; Hu, Juejun

    2017-09-01

    Recent development of a novel micro-scale PV/CPV technology is presented. The Wafer Integrated Micro-scale PV approach (WPV) seamlessly integrates multijunction micro-cells with a multi-functional silicon platform that provides optical micro-concentration, hybrid photovoltaic, and mechanical micro-assembly. The wafer-embedded micro-concentrating elements is shown to considerably improve the concentration-acceptance-angle product, potentially leading to dramatically reduced module materials and fabrication costs, sufficient angular tolerance for low-cost trackers, and an ultra-compact optical architecture, which makes the WPV module compatible with commercial flat panel infrastructures. The PV/CPV hybrid architecture further allows the collection of both direct and diffuse sunlight, thus extending the geographic and market domains for cost-effective PV system deployment. The WPV approach can potentially benefits from both the high performance of multijunction cells and the low cost of flat plate Si PV systems.

  20. MicroRNA function in Drosophila melanogaster.

    Science.gov (United States)

    Carthew, Richard W; Agbu, Pamela; Giri, Ritika

    2017-05-01

    Over the last decade, microRNAs have emerged as critical regulators in the expression and function of animal genomes. This review article discusses the relationship between microRNA-mediated regulation and the biology of the fruit fly Drosophila melanogaster. We focus on the roles that microRNAs play in tissue growth, germ cell development, hormone action, and the development and activity of the central nervous system. We also discuss the ways in which microRNAs affect robustness. Many gene regulatory networks are robust; they are relatively insensitive to the precise values of reaction constants and concentrations of molecules acting within the networks. MicroRNAs involved in robustness appear to be nonessential under uniform conditions used in conventional laboratory experiments. However, the robust functions of microRNAs can be revealed when environmental or genetic variation otherwise has an impact on developmental outcomes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Micro plate fission chamber development

    International Nuclear Information System (INIS)

    Wang Mei; Wen Zhongwei; Lin Jufang; Jiang Li; Liu Rong; Wang Dalun

    2014-01-01

    To conduct the measurement of neutron flux and the fission rate distribution at several position in assemblies, the micro plate fission chamber was designed and fabricated. Since the requirement of smaller volume and less structure material was taken into consideration, it is convinient, commercial and practical to use fission chamber to measure neutron flux in specific condition. In this paper, the structure of fission chamber and process of fabrication were introduced and performance test result was presented. The detection efficiency is 91.7%. (authors)

  2. Northern micro-grid project

    Energy Technology Data Exchange (ETDEWEB)

    Curtis, David; Singh, Bob

    2010-09-15

    The electrical distribution system for the Kasabonika Lake First Nation in northern Ontario (Canada) consumed 1.2 million liters of diesel fuel in 2008, amounting to 3,434 tones of CO2 emissions. The Northern Micro-Grid Project, supported by seven partners, involves integrating renewable generation & storage into the Kasabonika Lake distribution system. Through R&D and demonstration, the objectives are to reduce the amount of diesel consumed, support the distribution system exclusively on renewable resources during light loads, engage and impart knowledge/training to better position the community for future opportunities. The paper will discuss challenges, opportunities and future plans associated with the project.

  3. Micro-optomechanical trampoline resonators

    Science.gov (United States)

    Pepper, Brian; Kleckner, Dustin; Sonin, Petro; Jeffrey, Evan; Bouwmeester, Dirk

    2011-03-01

    Recently, micro-optomechanical devices have been proposed for implementation of experiments ranging from non-demolition measurements of phonon number to creation of macroscopic quantum superpositions. All have strenuous requirements on optical finesse, mechanical quality factor, and temperature. We present a set of devices composed of dielectric mirrors on Si 3 N4 trampoline resonators. We describe the fabrication process and present data on finesse and quality factor. The authors gratefully acknowledge support from NSF PHY-0804177 and Marie Curie EXT-CT-2006-042580.

  4. Micro-transactions for concurrent data structures

    DEFF Research Database (Denmark)

    Meawad, Fadi; Iyer, Karthik; Schoeberl, Martin

    2013-01-01

    implementation of transactional memory that we call micro-transactions. In particular, we argue that hardware support for micro-transactions allows us to efficiently implement certain data structures. Those data structures are difficult to realize with the atomic operations provided by stock hardware and provide......, atomic instructions, and micro-transactions. Our results suggest that transactional memory is an interesting alternative to traditional concurrency control mechanisms....

  5. Fabrication of Micro Components by Electrochemical Deposition

    DEFF Research Database (Denmark)

    Tang, Peter Torben

    The main issue of this thesis is the combination of electrochemical deposition of metals and micro machining. Processes for electroplating and electroless plating of nickel and nickel alloys have been developed and optimised for compatibility with microelectronics and silicon based micromechanics...... of electrochemical machining and traditional machining is compared to micro machining techniques as performed in the field of microelectronics. Various practical solutions and equipment for electrochemical deposition of micro components are demonstrated, as well as the use and experience obtained utilising...

  6. Challenging the sustainability of micro products development

    DEFF Research Database (Denmark)

    De Grave, Arnaud; Olsen, Stig Irving

    2006-01-01

    Environmental aspects are one of the biggest concerns regarding the future of manufacturing and product development sustainability. Furthermore, micro products and micro technologies are often seen as the next big thing in terms of possible mass market trend and boom. Many questions are raised...... and the intermediate parts which can be in-process created. Possible future trends for micro products development scheme involving environmental concerns are given....

  7. Development of 3d micro-nano hybrid patterns using anodized aluminum and micro-indentation

    Energy Technology Data Exchange (ETDEWEB)

    Shin, Hong Gue; Kwon, Jong Tae [Division of Mechanical Engineering and Mechatronics, Kangwon National University, 1 Kangwondaehakgil, Chunchon, Gangwon-do, 200-701 (Korea, Republic of); Seo, Young Ho [Division of Mechanical Engineering and Mechatronics, Kangwon National University, 1 Kangwondaehakgil, Chunchon, Gangwon-do, 200-701 (Korea, Republic of)], E-mail: mems@kangwon.ac.kr; Kim, Byeong Hee [Division of Mechanical Engineering and Mechatronics, Kangwon National University, 1 Kangwondaehakgil, Chunchon, Gangwon-do, 200-701 (Korea, Republic of)

    2008-07-31

    We developed a simple and cost-effective method of fabricating 3D micro-nano hybrid patterns in which micro-indentation is applied on the anodized aluminum substrate. Nano-patterns were formed first on the aluminum substrate, and then micro-patterns were fabricated by deforming the nano-patterned aluminum substrate. Hemispherical nano-patterns with a 150 nm-diameter on an aluminum substrate were fabricated by anodizing and alumina removing process. Then, micro-pyramid patterns with a side-length of 50 {mu}m were formed on the nano-patterns using micro-indentation. To verify 3D micro-nano hybrid patterns, we replicated 3D micro-nano hybrid patterns by a hot-embossing process. 3D micro-nano hybrid patterns may be used in nano-photonic devices and nano-biochips applications.

  8. Development of 3d micro-nano hybrid patterns using anodized aluminum and micro-indentation

    International Nuclear Information System (INIS)

    Shin, Hong Gue; Kwon, Jong Tae; Seo, Young Ho; Kim, Byeong Hee

    2008-01-01

    We developed a simple and cost-effective method of fabricating 3D micro-nano hybrid patterns in which micro-indentation is applied on the anodized aluminum substrate. Nano-patterns were formed first on the aluminum substrate, and then micro-patterns were fabricated by deforming the nano-patterned aluminum substrate. Hemispherical nano-patterns with a 150 nm-diameter on an aluminum substrate were fabricated by anodizing and alumina removing process. Then, micro-pyramid patterns with a side-length of 50 μm were formed on the nano-patterns using micro-indentation. To verify 3D micro-nano hybrid patterns, we replicated 3D micro-nano hybrid patterns by a hot-embossing process. 3D micro-nano hybrid patterns may be used in nano-photonic devices and nano-biochips applications

  9. Photolithography and Micro-Fabrication/ Packaging Laboratories

    Data.gov (United States)

    Federal Laboratory Consortium — The Photolithography and Micro-Fabrication/Packaging laboratories provide research level semiconductor processing equipment and facilities that do not require a full...

  10. Levitating Micro-Actuators: A Review

    Directory of Open Access Journals (Sweden)

    Kirill V. Poletkin

    2018-04-01

    Full Text Available Through remote forces, levitating micro-actuators completely eliminate mechanical attachment between the stationary and moving parts of a micro-actuator, thus providing a fundamental solution to overcoming the domination of friction over inertial forces at the micro-scale. Eliminating the usual mechanical constraints promises micro-actuators with increased operational capabilities and low dissipation energy. Further reduction of friction and hence dissipation by means of vacuum leads to dramatic increases of performance when compared to mechanically tethered counterparts. In order to efficiently employ the benefits provided by levitation, micro-actuators are classified according to their physical principles as well as by their combinations. Different operating principles, structures, materials and fabrication methods are considered. A detailed analysis of the significant achievements in the technology of micro-optics, micro-magnets and micro-coil fabrication, along with the development of new magnetic materials during recent decades, which has driven the creation of new application domains for levitating micro-actuators is performed.

  11. Micro-Lid For Sealing Sample Reservoirs of micro-Extraction Systems

    Data.gov (United States)

    National Aeronautics and Space Administration — We propose to develop a proof-of-concept micro-Lid (µLid) to tightly seal a micro-sampler or micro-extraction system. Fabrication of µLid would be conducted in the...

  12. [Researches on biomechanics of micro-implant-bone interface and optimum design of micro implant's neck].

    Science.gov (United States)

    Deng, Feng; Zhang, Lei; Zhang, Yi; Song, Jin-lin; Fan, Yuboa

    2007-07-01

    To compare and analyze the stress distribution at the micro-implant-bone interface based on the different micro-implant-bone conditioned under orthodontic load, and to optimize the design of micro implant's neck. An adult skull with all tooth was scanned by spiral CT, and the data were imported into computer for three-dimensional reconstruction with software Mimics 9.0. The three dimensional finite element models of three micro-implant-bone interfaces(initial stability, full osseointegration and fibrous integration) were analyzed by finite element analysis software ABAQUS6.5. The primary stress distributions of different micro-implant-bone conditions were evaluated when 2N force was loaded. Then the diameter less than 1.5 mm of the micro implant's neck was added with 0.2 mm, to compare the stress distribution of the modified micro-implant-bone interface with traditional type. The stress mostly concentrated on the neck of micro implant and the full osseointegration interface in all models showed the lowest strain level. Compared with the traditional type, the increasing diameter neck of the micro implant obviously decreased the stress level in all the three conditions. The micro-implant-bone interface and the diameter of micro implant's neck both are the important influence factors to the stress distribution of micro implant.

  13. Integrated Micro Product and Technology Development

    DEFF Research Database (Denmark)

    Hansen, Hans Nørgaard

    2003-01-01

    The paper addresses the issues of integrated micro product and technology development. The implications of the decisions in the design phase on the subsequent manufacturing processes are considered vital. A coherent process chain is a necessary prerequisite for the realisation of the industrial...... potential of micro technology....

  14. Can micro-volunteering help in Africa?

    CSIR Research Space (South Africa)

    Butgereit, L

    2013-05-01

    Full Text Available is convenient to the micro-volunteer, and in small pieces of time (bitesized). This paper looks at a micro-volunteering project where participants can volunteer for five to ten minutes at a time using a smart phone and assist pupils with their mathematics....

  15. Micro-incubator for bacterial biosensing applications

    CSIR Research Space (South Africa)

    Clasen, E

    2016-09-01

    Full Text Available The presence of Escherichia coli (E. coli) is a commonly used indicator micro-organism to determine whether water is safe for human consumption. This paper discusses the design of a micro-incubator that can be applied to concentrate bacteria prior...

  16. A novel differential frequency micro-gyroscope

    KAUST Repository

    Nayfeh, A. H.; Abdel-Rahman, E. M.; Ghommem, M.

    2013-01-01

    We present a frequency-domain method to measure angular speeds using electrostatic micro-electro-mechanical system actuators. Towards this end, we study a single-axis gyroscope made of a micro-cantilever and a proof-mass coupled to two fixed

  17. Wafer-scale micro-optics fabrication

    Science.gov (United States)

    Voelkel, Reinhard

    2012-07-01

    Micro-optics is an indispensable key enabling technology for many products and applications today. Probably the most prestigious examples are the diffractive light shaping elements used in high-end DUV lithography steppers. Highly-efficient refractive and diffractive micro-optical elements are used for precise beam and pupil shaping. Micro-optics had a major impact on the reduction of aberrations and diffraction effects in projection lithography, allowing a resolution enhancement from 250 nm to 45 nm within the past decade. Micro-optics also plays a decisive role in medical devices (endoscopes, ophthalmology), in all laser-based devices and fiber communication networks, bringing high-speed internet to our homes. Even our modern smart phones contain a variety of micro-optical elements. For example, LED flash light shaping elements, the secondary camera, ambient light and proximity sensors. Wherever light is involved, micro-optics offers the chance to further miniaturize a device, to improve its performance, or to reduce manufacturing and packaging costs. Wafer-scale micro-optics fabrication is based on technology established by the semiconductor industry. Thousands of components are fabricated in parallel on a wafer. This review paper recapitulates major steps and inventions in wafer-scale micro-optics technology. The state-of-the-art of fabrication, testing and packaging technology is summarized.

  18. Addressing Youth Employment Through Micro- and Small ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    ... Employment Through Micro- and Small-Enterprise Development in Ethiopia. Youth unemployment has emerged as a key challenge facing developing and ... to youth to start small businesses and to youth-led micro- and small-enterprises. ... the barriers and challenges young Ethiopian men and women face in the labour ...

  19. A thermally self-sustained micro-power plant with integrated micro-solid oxide fuel cells, micro-reformer and functional micro-fluidic carrier

    Science.gov (United States)

    Scherrer, Barbara; Evans, Anna; Santis-Alvarez, Alejandro J.; Jiang, Bo; Martynczuk, Julia; Galinski, Henning; Nabavi, Majid; Prestat, Michel; Tölke, René; Bieberle-Hütter, Anja; Poulikakos, Dimos; Muralt, Paul; Niedermann, Philippe; Dommann, Alex; Maeder, Thomas; Heeb, Peter; Straessle, Valentin; Muller, Claude; Gauckler, Ludwig J.

    2014-07-01

    Low temperature micro-solid oxide fuel cell (micro-SOFC) systems are an attractive alternative power source for small-size portable electronic devices due to their high energy efficiency and density. Here, we report on a thermally self-sustainable reformer-micro-SOFC assembly. The device consists of a micro-reformer bonded to a silicon chip containing 30 micro-SOFC membranes and a functional glass carrier with gas channels and screen-printed heaters for start-up. Thermal independence of the device from the externally powered heater is achieved by exothermic reforming reactions above 470 °C. The reforming reaction and the fuel gas flow rate of the n-butane/air gas mixture controls the operation temperature and gas composition on the micro-SOFC membrane. In the temperature range between 505 °C and 570 °C, the gas composition after the micro-reformer consists of 12 vol.% to 28 vol.% H2. An open-circuit voltage of 1.0 V and maximum power density of 47 mW cm-2 at 565 °C is achieved with the on-chip produced hydrogen at the micro-SOFC membranes.

  20. Classification of assembly techniques for micro products

    DEFF Research Database (Denmark)

    Hansen, Hans Nørgaard; Tosello, Guido; Gegeckaite, Asta

    2005-01-01

    of components and level of integration are made. This paper describes a systematic characterization of micro assembly methods. This methodology offers the opportunity of a cross comparison among different techniques to gain a choosing principle of the favourable micro assembly technology in a specific case...

  1. Micromachining of buried micro channels in silicon

    NARCIS (Netherlands)

    de Boer, Meint J.; Tjerkstra, R.W.; Berenschot, Johan W.; Jansen, Henricus V.; Burger, G.J.; Burger, G.J.; Gardeniers, Johannes G.E.; Elwenspoek, Michael Curt; van den Berg, Albert

    A new method for the fabrication of micro structures for fluidic applications, such as channels, cavities, and connector holes in the bulk of silicon wafers, called buried channel technology (BCT), is presented in this paper. The micro structures are constructed by trench etching, coating of the

  2. Investigation on the micro injection molding process of an overmolded multi-material micro component

    DEFF Research Database (Denmark)

    Baruffi, Federico; Calaon, Matteo; Tosello, Guido

    and difficult assembly steps, being the plastic molded directly on a metal substrate. In this scenario, an investigation on the fully automated micro overmolding manufacturing technology of a three-material micro component for acoustic applications has been carried out. Preliminary experiments allowed......Micro injection molding (μIM) is one of the few technologies capable of meeting the increasing demand of complex shaped micro plastic parts. This process, combined with the overmolding technique, allows a fast and cost-efficient production of multi-material micro components, saving numerous...

  3. MicroRNAs and Presbycusis.

    Science.gov (United States)

    Hu, Weiming; Wu, Junwu; Jiang, Wenjing; Tang, Jianguo

    2018-02-01

    Presbycusis (age-related hearing loss) is the most universal sensory degenerative disease in elderly people caused by the degeneration of cochlear cells. Non-coding microRNAs (miRNAs) play a fundamental role in gene regulation in almost every multicellular organism, and control the aging processes. It has been identified that various miRNAs are up- or down-regulated during mammalian aging processes in tissue-specific manners. Most miRNAs bind to specific sites on their target messenger-RNAs (mRNAs) and decrease their expression. Germline mutation may lead to dysregulation of potential miRNAs expression, causing progressive hair cell degeneration and age-related hearing loss. Therapeutic innovations could emerge from a better understanding of diverse function of miRNAs in presbycusis. This review summarizes the relationship between miRNAs and presbycusis, and presents novel miRNAs-targeted strategies against presbycusis.

  4. An electromagnetic micro-undulator

    International Nuclear Information System (INIS)

    Nassiri, A.; Turner, L.R.

    1997-01-01

    Microfabrication technology using the LIGA (a German acronym for Lithography, Electroforming, and Molding) process offers an attractive alternative for fabricating precision devices with micron-sized features. One such device is a mm-sized micro-undulator with potential applications in a table-top synchrotron light source for medical and other industrial uses. The undulator consists of a silver conductor embedded in poles and substrate of nickel-iron. Electromagnetic modeling of the undulator is done using the eddy current computer code ELEKTRA. Computations predict a field pattern of appropriate strength and quality if the current can be prevented from being shunted from silver by the nickel-iron poles either through insulation or through slotted poles. The design of the undulator along with the computational results are discussed

  5. Small proteins in cyanobacteria provide a paradigm for the functional analysis of the bacterial micro-proteome.

    Science.gov (United States)

    Baumgartner, Desiree; Kopf, Matthias; Klähn, Stephan; Steglich, Claudia; Hess, Wolfgang R

    2016-11-28

    Despite their versatile functions in multimeric protein complexes, in the modification of enzymatic activities, intercellular communication or regulatory processes, proteins shorter than 80 amino acids (μ-proteins) are a systematically underestimated class of gene products in bacteria. Photosynthetic cyanobacteria provide a paradigm for small protein functions due to extensive work on the photosynthetic apparatus that led to the functional characterization of 19 small proteins of less than 50 amino acids. In analogy, previously unstudied small ORFs with similar degrees of conservation might encode small proteins of high relevance also in other functional contexts. Here we used comparative transcriptomic information available for two model cyanobacteria, Synechocystis sp. PCC 6803 and Synechocystis sp. PCC 6714 for the prediction of small ORFs. We found 293 transcriptional units containing candidate small ORFs ≤80 codons in Synechocystis sp. PCC 6803, also including the known mRNAs encoding small proteins of the photosynthetic apparatus. From these transcriptional units, 146 are shared between the two strains, 42 are shared with the higher plant Arabidopsis thaliana and 25 with E. coli. To verify the existence of the respective μ-proteins in vivo, we selected five genes as examples to which a FLAG tag sequence was added and re-introduced them into Synechocystis sp. PCC 6803. These were the previously annotated gene ssr1169, two newly defined genes norf1 and norf4, as well as nsiR6 (nitrogen stress-induced RNA 6) and hliR1(high light-inducible RNA 1) , which originally were considered non-coding. Upon activation of expression via the Cu 2+. responsive petE promoter or from the native promoters, all five proteins were detected in Western blot experiments. The distribution and conservation of these five genes as well as their regulation of expression and the physico-chemical properties of the encoded proteins underline the likely great bandwidth of small protein

  6. MicroComputed Tomography: Methodology and Applications

    International Nuclear Information System (INIS)

    Stock, Stuart R.

    2009-01-01

    Due to the availability of commercial laboratory systems and the emergence of user facilities at synchrotron radiation sources, studies of microcomputed tomography or microCT have increased exponentially. MicroComputed Technology provides a complete introduction to the technology, describing how to use it effectively and understand its results. The first part of the book focuses on methodology, covering experimental methods, data analysis, and visualization approaches. The second part addresses various microCT applications, including porous solids, microstructural evolution, soft tissue studies, multimode studies, and indirect analyses. The author presents a sufficient amount of fundamental material so that those new to the field can develop a relative understanding of how to design their own microCT studies. One of the first full-length references dedicated to microCT, this book provides an accessible introduction to field, supplemented with application examples and color images.

  7. Biologically Inspired Micro-Flight Research

    Science.gov (United States)

    Raney, David L.; Waszak, Martin R.

    2003-01-01

    Natural fliers demonstrate a diverse array of flight capabilities, many of which are poorly understood. NASA has established a research project to explore and exploit flight technologies inspired by biological systems. One part of this project focuses on dynamic modeling and control of micro aerial vehicles that incorporate flexible wing structures inspired by natural fliers such as insects, hummingbirds and bats. With a vast number of potential civil and military applications, micro aerial vehicles represent an emerging sector of the aerospace market. This paper describes an ongoing research activity in which mechanization and control concepts for biologically inspired micro aerial vehicles are being explored. Research activities focusing on a flexible fixed- wing micro aerial vehicle design and a flapping-based micro aerial vehicle concept are presented.

  8. MicroRNA involvement in glioblastoma pathogenesis

    International Nuclear Information System (INIS)

    Novakova, Jana; Slaby, Ondrej; Vyzula, Rostislav; Michalek, Jaroslav

    2009-01-01

    MicroRNAs are endogenously expressed regulatory noncoding RNAs. Altered expression levels of several microRNAs have been observed in glioblastomas. Functions and direct mRNA targets for these microRNAs have been relatively well studied over the last years. According to these data, it is now evident, that impairment of microRNA regulatory network is one of the key mechanisms in glioblastoma pathogenesis. MicroRNA deregulation is involved in processes such as cell proliferation, apoptosis, cell cycle regulation, invasion, glioma stem cell behavior, and angiogenesis. In this review, we summarize the current knowledge of miRNA functions in glioblastoma with an emphasis on its significance in glioblastoma oncogenic signaling and its potential to serve as a disease biomarker and a novel therapeutic target in oncology.

  9. Precision moulding of polymer micro components

    DEFF Research Database (Denmark)

    Tosello, Guido

    2008-01-01

    The present research work contains a study concerning polymer micro components manufacturing by means of the micro injection moulding (µIM) process. The overall process chain was considered and investigated during the project, including part design and simulation, tooling, process analysis, part...... optimization, quality control, multi-material solutions. A series of experimental investigations were carried out on the influence of the main µIM process factors on the polymer melt flow within micro cavities. These investigations were conducted on a conventional injection moulding machine adapted...... to the production of micro polymer components, as well as on a micro injection moulding machine. A new approach based on coordinate optical measurement of flow markers was developed during the project for the characterization of the melt flow. In-line pressure measurements were also performed to characterize...

  10. Micro-Scale Regenerative Heat Exchanger

    Science.gov (United States)

    Moran, Matthew E.; Stelter, Stephan; Stelter, Manfred

    2004-01-01

    A micro-scale regenerative heat exchanger has been designed, optimized and fabricated for use in a micro-Stirling device. Novel design and fabrication techniques enabled the minimization of axial heat conduction losses and pressure drop, while maximizing thermal regenerative performance. The fabricated prototype is comprised of ten separate assembled layers of alternating metal-dielectric composite. Each layer is offset to minimize conduction losses and maximize heat transfer by boundary layer disruption. A grating pattern of 100 micron square non-contiguous flow passages were formed with a nominal 20 micron wall thickness, and an overall assembled ten-layer thickness of 900 microns. Application of the micro heat exchanger is envisioned in the areas of micro-refrigerators/coolers, micropower devices, and micro-fluidic devices.

  11. Laser beam micro-milling of micro-channels in aerospace alloys

    CERN Document Server

    Ahmed, Naveed; Al-Ahmari, Abdulrahman

    2017-01-01

    This volume is greatly helpful to micro-machining and laser engineers as it offers obliging guidelines about the micro-channel fabrications through Nd:YAG laser beam micro-milling. The book also demonstrates how the laser beam micro-milling behaves when operating under wet conditions (under water), and explores what are the pros and cons of this hybrid technique. From the predictive mathematical models, the readers can easily estimate the resulting micro-channel size against the desired laser parametric combinations. The book considers micro-channels in three highly important research materials commonly used in aerospace industry: titanium alloy Ti-6Al-4V, nickel alloy Inconel 718 and aluminum alloy AA 2024. Therefore, the book is highly practicable in the fields of micro-channel heat exchangers, micro-channel aerospace turbine blades, micro-channel heat pipes, micro-coolers and micro-channel pulsating heat plates. These are frequently used in various industries such as aerospace, automotive, biomedical and m...

  12. Three-dimensional micro assembly of a hinged nickel micro device by magnetic lifting and micro resistance welding

    International Nuclear Information System (INIS)

    Chang, Chun-Wei; Hsu, Wensyang

    2009-01-01

    The three-dimensional micro assembly of hinged nickel micro devices by magnetic lifting and micro resistance welding is proposed here. By an electroplating-based surface machining process, the released nickel structure with the hinge mechanism can be fabricated. Lifting of the released micro structure to different tilted angles is accomplished by controlling the positions of a magnet beneath the device. An in situ electro-thermal actuator is used here to provide the pressing force in micro resistance welding for immobilizing the tilted structure. The proposed technique is shown to immobilize micro devices at controlled angles ranging from 14° to 90° with respect to the substrate. Design parameters such as the electro-thermal actuator and welding beam width are also investigated. It is found that there is a trade-off in beam width design between large contact pressure and low thermal deformation. Different dominated effects from resistivity enhancement and contact area enlargement during the welding process are also observed in the dynamic resistance curves. Finally, a lifted and immobilized electro-thermal bent-beam actuator is shown to displace upward about 27.7 µm with 0.56 W power input to demonstrate the capability of electrical transmission at welded joints by the proposed 3D micro assembly technique

  13. Manufacturing and application of micro computer for control

    International Nuclear Information System (INIS)

    Park, Seung Man; Heo, Gyeong; Yun, Jun Young

    1990-05-01

    This book deals with machine code and assembly program for micro computer. It composed of 20 chapters, which are micro computer system, practice of a storage cell, manufacturing 1 of micro computer, manufacturing 2 of micro computer, manufacturing of micro computer AID-80A, making of machine language, interface like Z80-PIO and 8255A(PPI), counter and timer interface, exercise of basic command, arithmetic operation, arrangement operation, an indicator control, music playing, detection of input of PIO. control of LED of PIO, PIO mode, CTC control by micro computer, SIO control by micro computer and application by micro computer.

  14. Micro-incubator for bacterial biosensing applications

    Science.gov (United States)

    Clasen, Estine; Land, Kevin; Joubert, Trudi-Heleen

    2016-02-01

    The presence of Escherichia coli (E. coli ) is a commonly used indicator micro-organism to determine whether water is safe for human consumption.1 This paper discusses the design of a micro-incubator that can be applied to concentrate bacteria prior to environmental water quality screening tests. High sensitivity and rapid test time is essential and there is a great need for these tests to be implemented on-site without the use of a laboratory infrastructure. In the light of these requirements, a mobile micro-incubator was designed, manufactured and characterised. A polydimethylsiloxane (PDMS) receptacle has been designed to house the 1-5 ml cell culture sample.2 A nano-silver printed electronics micro-heater has been designed to incubate the bacterial sample, with an array of temperature sensors implemented to accurately measure the sample temperature at various locations in the cell culture well. The micro-incubator limits the incubation temperature range to 37+/-3 °C in order to ensure near optimal growth of the bacteria at all times.3 The incubation time is adjustable between 30 minutes and 9 hours with a maximum rise time of 15 minutes to reach the set-point temperature. The surface area of the printed nano silver heating element is 500 mm2. Electrical and COMSOL Multiphysics simulations are included in order to give insight on micro-incubator temperature control. The design and characterization of this micro-incubator allows for further research in biosensing applications.

  15. Updating the Micro-Tom TILLING platform.

    Science.gov (United States)

    Okabe, Yoshihiro; Ariizumi, Tohru; Ezura, Hiroshi

    2013-03-01

    The dwarf tomato variety Micro-Tom is regarded as a model system for functional genomics studies in tomato. Various tomato genomic tools in the genetic background of Micro-Tom have been established, such as mutant collections, genome information and a metabolomic database. Recent advances in tomato genome sequencing have brought about a significant need for reverse genetics tools that are accessible to the larger community, because a great number of gene sequences have become available from public databases. To meet the requests from the tomato research community, we have developed the Micro-Tom Targeting-Induced Local Lesions IN Genomes (TILLING) platform, which is comprised of more than 5000 EMS-mutagenized lines. The platform serves as a reverse genetics tool for efficiently identifying mutant alleles in parallel with the development of Micro-Tom mutant collections. The combination of Micro-Tom mutant libraries and the TILLING approach enables researchers to accelerate the isolation of desirable mutants for unraveling gene function or breeding. To upgrade the genomic tool of Micro-Tom, the development of a new mutagenized population is underway. In this paper, the current status of the Micro-Tom TILLING platform and its future prospects are described.

  16. Fabrication of micro metallic valve and pump

    Science.gov (United States)

    Yang, Ming; Kabasawa, Yasunari; Ito, Kuniyoshi

    2010-03-01

    Fabrication of micro devices by using micro metal forming was proposed by the authors. We developed a desktop servo-press machine with precise tooling system. Precise press forming processes including micro forging and micro joining has been carried out in a progressive die. In this study, micro metallic valve and pump were fabricated by using the precise press forming. The components are made of sheet metals, and assembled in to a unit in the progressive die. A micro check-valve with a diameter of 3mm and a length of 3.2mm was fabricated, and the property of flow resistance was evaluated. The results show that the check valve has high property of leakage proof. Since the valve is a unit parts with dimensions of several millimeters, it has advantage to be adapted to various pump design. Here, two kinds of micro pumps with the check-valves were fabricated. One is diaphragm pump actuated by vibration of the diaphragm, and another is tube-shaped pump actuated by resonation. The flow quantities of the pumps were evaluated and the results show that both of the pumps have high pumping performance.

  17. Aluminum Templates of Different Sizes with Micro-, Nano- and Micro/Nano-Structures for Cell Culture

    Directory of Open Access Journals (Sweden)

    Ming-Liang Yen

    2017-10-01

    Full Text Available This study investigates the results of cell cultures on aluminum (Al templates with flat-structures, micro-structures, nano-structures and micro/nano-structures. An Al template with flat-structure was obtained by electrolytic polishing; an Al template with micro-structure was obtained by micro-powder blasting; an Al template with nano-structure was obtained by aluminum anodization; and an Al template with micro/nano-structure was obtained by micro-powder blasting and then anodization. Osteoblast-like cells were cultured on aluminum templates with various structures. The microculture tetrazolium test assay was utilized to assess the adhesion, elongation, and proliferation behaviors of cultured osteoblast-like cells on aluminum templates with flat-structures, micro-structures, nano-structures, and micro/nano-structures. The results showed that the surface characterization of micro/nano-structure of aluminum templates had superhydrophilic property, and these also revealed that an aluminum template with micro/nano-structure could provide the most suitable growth situation for cell culture.

  18. Formability of Micro-Tubes in Hydroforming

    International Nuclear Information System (INIS)

    Hartl, Christoph; Anyasodor, Gerald; Lungershausen, Joern

    2011-01-01

    Micro-hydroforming is a down-scaled metal forming process, based on the expansion of micro-tubes by internal pressurization within a die cavity. The objective of micro-hydroforming is to provide a technology for the economic mass production of complex shaped hollow micro-components. Influence of size effects in metal forming processes increases with scaling down of metal parts. Investigations into the change in formability of micro-tubes due to metal part scaling down constituted an important subject within the conducted fundamental research work. Experimental results are presented, concerning the analysis of the formability of micro-tubes made from stainless steel AISI 304 with an outer diameter of 800 μm and a wall thickness of 40 μm. An average ratio of tube wall thickness to grain size of 1.54 of up to 2.56 was analyzed. Miniaturised mechanical standard methods as well as bulge tests with internal hydrostatic pressurization of the tubular specimens were applied to analyze the influence of size-dependent effects. A test device was developed for the bulge experiments which enabled the pressurization of micro-tubes with internal pressures up to 4000 bar. To determine the attainable maximum achievable expansion ratio the tubes were pressurized in the bulge tests with increasing internal pressure until instability due to necking and subsequent bursting occurred. Comparisons with corresponding tests of macro-tubes, made from the here investigated material, showed a change in formability of micro-tubes which was attributed to the scaling down of the hydroforming process. In addition, a restricted applicability of existing theoretical correlations for the determination of the maximum pressure at bursting was observed for down-scaled micro-hydroforming.

  19. Progress in micro-pattern gas detectors

    International Nuclear Information System (INIS)

    Bellazzini, Ronaldo

    2001-01-01

    Micro-Pattern Gas Detectors are position-sensitive proportional counters whose sense electrodes are constructed using micro-electronics , thin-film or advanced PCB techniques.The feature size attainable using these methods is of the order of a few microns and the detectors demonstrate excellent spatial resolution and fast charge collection. I will review recent progress on Micro patterned Gas Detectors for tracking and other cross-disciplinary applications.I will focus on the design principles,performance capability and limitations. A short list of interesting applications will be discussed

  20. Micro Nano Replication Processes and Applications

    CERN Document Server

    Kang, Shinill

    2012-01-01

    This book is an introduction to the fundamentals and processes for micro and nano molding for plastic components. In addition to the basics, the book covers applications details and examples. The book helps both students and professionals to understand and work with the growing tools of molding and uses for micro and nano-sized plastic parts.Provides a comprehensive presentation on fundamentals and practices of manufacturing for micro / nano sized plastics partsCovers a relatively new but fast-growing field that is impacting any industry using plastic parts in their products (electronics, tele

  1. Neutrino Interactions in MicroBooNE

    OpenAIRE

    Del Tutto, Marco

    2017-01-01

    MicroBooNE is a liquid-argon-based neutrino experiment, which began collecting data in Fermilab's Booster neutrino beam in October 2015. Physics goals of the experiment include probing the source of the anomalous excess of electron-like events in MiniBooNE. In addition to this, MicroBooNE is carrying out an extensive cross section physics program that will help to probe current theories on neutrino-nucleon interactions and nuclear effects. These proceedings summarise the status of MicroBooNE'...

  2. MR of pituitary micro-adenomas

    International Nuclear Information System (INIS)

    Le Marec, E.; Ait Ameur, A.; David, H.; Pharaboz, C.

    1997-01-01

    Most of the time, rationales to look for pituitary micro-adenomas are based on endocrinal disorder. MRI is often helpful to confirm diagnosis. It gives information about micro-adenomas size and localisation. If conventional sequence are inadequate, a dynamic sequence has then to be performed after Gadolinium injection. Any disorder observed from the pituitary gland must be correlated with the clinical observation and results from biochemistry analysis. False positive happens quite open because of gland morphological variation, incidentalomas and partial volumes. MRI offers the possibility to follow-up treated micro-adenomas evolution especially to detect recurrence. (author)

  3. A Micro-Grid Battery Storage Management

    DEFF Research Database (Denmark)

    Mahat, Pukar; Escribano Jiménez, Jorge; Moldes, Eloy Rodríguez

    2013-01-01

    An increase in number of distributed generation (DG) units in power system allows the possibility of setting-up and operating micro-grids. In addition to a number of technical advantages, micro-grid operation can also reduce running costs by optimally scheduling the generation and/or storage...... systems under its administration. This paper presents an optimized scheduling of a micro-grid battery storage system that takes into account the next-day forecasted load and generation profiles and spot electricity prices. Simulation results show that the battery system can be scheduled close to optimal...

  4. MICRO AUTO GASIFICATION SYSTEM: EMISSIONS ...

    Science.gov (United States)

    A compact, CONEX-housed waste to energy unit, Micro Auto Gasification System (MAGS), was characterized for air emissions from burning of military waste types. The MAGS unit is a dual chamber gasifier with a secondary diesel-fired combustor. Eight tests were conducted with multiple waste types in a 7-day period at the Kilauea Military Camp in Hawai’i. The emissions characterized were chosen based on regulatory emissions limits as well as their ability to cause adverse health effects on humans: particulate matter (PM), mercury, heavy metals, volatile organic compounds (VOCs), polyaromatic hydrocarbons (PAHs), and polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs). Three military waste feedstock compositions reflecting the variety of wastes to be encountered in theatre were investigated: standard waste (SW), standard waste with increased plastic content (HP), standard waste without SW food components but added first strike ration (FSR) food and packaging material (termed FSR). A fourth waste was collected from the Kilauea dumpster that served the dining facility and room lodging (KMC). Limited scrubber water and solid ash residue samples were collected to obtain a preliminary characterization of these effluents/residues.Gasifying SW, HP, and KMC resulted in similar PCDD/PCDF stack concentrations, 0.26-0.27 ng TEQ/m3 at 7% O2, while FSR waste generated a notably higher stack concentration of 0.68 ng TEQ/m3 at 7% O2. The PM emission

  5. An on-line monitoring system for a micro electrical discharge machining (micro-EDM) process

    International Nuclear Information System (INIS)

    Liao, Y S; Chang, T Y; Chuang, T J

    2008-01-01

    A pulse-type discriminating system to monitor the process of micro electrical discharge machining (micro-EDM) is developed and implemented. The specific features are extracted and the pulses from a RC-type power source are classified into normal, effective arc, transient short circuit and complex types. An approach to discriminate the pulse type according to three durations measured at three pre-determined voltage levels of a pulse is proposed. The developed system is verified by using simulated signals. Discrimination of the pulse trains in actual machining processes shows that the pulses are mainly the normal type for micro wire-EDM and micro-EDM milling. The pulse-type distribution varies during the micro-EDM drilling process. The percentage of complex-type pulse increases monotonically with the drilling depth. It starts to drop when the gap condition is seriously deteriorated. Accordingly, an on-line monitoring strategy for the micro-EDM drilling process is proposed

  6. Modeling High Pressure Micro Hollow Cathode Discharges

    National Research Council Canada - National Science Library

    Boeuf, Jean-Pierre; Pitchford, Leanne

    2004-01-01

    This report results from a contract tasking CPAT as follows: The Grantee will perform theoretical modeling of point, surface, and volume high-pressure plasmas created using Micro Hollow Cathode Discharge sources...

  7. The development of micro-gyroscope technology

    International Nuclear Information System (INIS)

    Liu, Kai; Zhang, Weiping; Chen, Wenyuan; Li, Kai; Dai, Fuyan; Cui, Feng; Wu, Xiaosheng; Ma, Gaoyin; Xiao, Qijun

    2009-01-01

    This review reports an overview and development of micro-gyroscope. The review first presents different types of micro-gyroscopes. Micro-gyroscopes in this review are categorized into Coriolis gyroscope, levitated rotor gyroscope, Sagnac gyroscope, nuclear magnetic resonance (NMR) gyroscope according to the working principle. Different principles, structures, materials, fabrications and control technologies of micro-gyroscopes are analyzed. This review compares different classes of gyroscopes in the aspects such as fabrication method, detection axis, materials, size and so on. Finally, the review evaluates the key technologies on how to improve the precision and anti-jamming ability and to extend the available applications of the gyroscopes in the market and patents as well. (topical review)

  8. Micro Resistojet for Small Satellites, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — Micro-resistojets offer an excellent combination of simplicity, performance and wet system mass for small satellites (<100 kg, <50 watts) requiring mN level...

  9. MicroCar 2003. Abstracts of papers

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2003-07-01

    Mechatronics for automotive applications is an important trend, combining mechanics, electronics and information technology. Micro- and nanomechatronics, particularly innovative research disciplines, will help create, in combination with advanced solutions from microsystem technologies, e.g. the electronic packaging, a range of entirely new developments in automobiles. Under the umbrella of the Automotive Suppliers Fair Z2003, it is happening for the very first time now that a momentous scientific conference, such as the MicroCar 2003, brings together representatives from car manufacturers and the electronics industry, as well as a large number of experts from technical colleges, universities and research institutes to discuss the new potentials and possibilities presented by the use of micro and nanomaterials in Leipzig. With the presentation of 50 papers, speakers will inform about their latest research results as well as current trends in micro and nanotechnologies for automotives. This issue is publishing the abstracts of this scientific event.

  10. Micro- and nanodevices integrated with biomolecular probes.

    Science.gov (United States)

    Alapan, Yunus; Icoz, Kutay; Gurkan, Umut A

    2015-12-01

    Understanding how biomolecules, proteins and cells interact with their surroundings and other biological entities has become the fundamental design criterion for most biomedical micro- and nanodevices. Advances in biology, medicine, and nanofabrication technologies complement each other and allow us to engineer new tools based on biomolecules utilized as probes. Engineered micro/nanosystems and biomolecules in nature have remarkably robust compatibility in terms of function, size, and physical properties. This article presents the state of the art in micro- and nanoscale devices designed and fabricated with biomolecular probes as their vital constituents. General design and fabrication concepts are presented and three major platform technologies are highlighted: microcantilevers, micro/nanopillars, and microfluidics. Overview of each technology, typical fabrication details, and application areas are presented by emphasizing significant achievements, current challenges, and future opportunities. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Micro Resistojet for Small Satellites, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — Micro-resistojets offer the best combination of simplicity, performance, wet system mass and power consumption for small satellites (<100kg, <50Watts)...

  12. Micro benchtop optics by bulk silicon micromachining

    Science.gov (United States)

    Lee, Abraham P.; Pocha, Michael D.; McConaghy, Charles F.; Deri, Robert J.

    2000-01-01

    Micromachining of bulk silicon utilizing the parallel etching characteristics of bulk silicon and integrating the parallel etch planes of silicon with silicon wafer bonding and impurity doping, enables the fabrication of on-chip optics with in situ aligned etched grooves for optical fibers, micro-lenses, photodiodes, and laser diodes. Other optical components that can be microfabricated and integrated include semi-transparent beam splitters, micro-optical scanners, pinholes, optical gratings, micro-optical filters, etc. Micromachining of bulk silicon utilizing the parallel etching characteristics thereof can be utilized to develop miniaturization of bio-instrumentation such as wavelength monitoring by fluorescence spectrometers, and other miniaturized optical systems such as Fabry-Perot interferometry for filtering of wavelengths, tunable cavity lasers, micro-holography modules, and wavelength splitters for optical communication systems.

  13. Hollow Micro-/Nanostructures: Synthesis and Applications

    KAUST Repository

    Lou, Xiong Wen (David); Archer, Lynden A.; Yang, Zichao

    2008-01-01

    for Portland cement, to produce concrete with enhanced strength and durability. This review is devoted to the progress made in the last decade in synthesis and applications of hollow micro-nanostructures. We present a comprehensive overview of synthetic

  14. Aerial photogrammetry procedure optimized for micro uav

    Directory of Open Access Journals (Sweden)

    T. Anai

    2014-06-01

    Full Text Available This paper proposes the automatic aerial photogrammetry procedure optimized for Micro UAV that has ability of autonomous flight. The most important goal of our proposed method is the reducing the processing cost for fully automatic reconstruction of DSM from a large amount of image obtained from Micro UAV. For this goal, we have developed automatic corresponding point generation procedure using feature point tracking algorithm considering position and attitude information, which obtained from onboard GPS-IMU integrated on Micro UAV. In addition, we have developed the automatic exterior orientation and registration procedure from the automatic generated corresponding points on each image and position and attitude information from Micro UAV. Moreover, in order to reconstruct precise DSM, we have developed the area base matching process which considering edge information. In this paper, we describe processing flow of our automatic aerial photogrammetry. Moreover, the accuracy assessment is also described. Furthermore, some application of automatic reconstruction of DSM will be desired.

  15. Micro Learning: A Modernized Education System

    Directory of Open Access Journals (Sweden)

    Omer Jomah

    2016-03-01

    Full Text Available Learning is an understanding of how the human brain is wired to learning rather than to an approach or a system. It is one of the best and most frequent approaches for the 21st century learners. Micro learning is more interesting due to its way of teaching and learning the content in a small, very specific burst. Here the learners decide what and when to learn. Content, time, curriculum, form, process, mediality, and learning type are the dimensions of micro learning. Our paper will discuss about micro learning and about the micro-content management system. The study will reflect the views of different users, and will analyze the collected data. Finally, it will be concluded with its pros and cons. 

  16. A CAMAC and FASTBUS engineering test environment supported by a MicroVAX/MicroVMS system

    International Nuclear Information System (INIS)

    Logg, C.A.

    1987-10-01

    A flexible, multiuser engineering test environment has been established for the engineers in SLAC's Electronic Instrumentation Engineering group. The system hardware includes a standard MicroVAX II and MicroVAX I with multiple CAMAC, FASTBUS, and GPIB instrumentation buses. The system software components include MicroVMS licenses with DECNET/SLACNET, FORTRAN, PASCAL, FORTH, and a versatile graphical display package. In addition, there are several software utilities available to facilitate FASTBUS and CAMAC prototype hardware debugging. 16 refs., 7 figs

  17. Micro Loudspeaker Behaviour versus 6½" Driver, Micro Loudspeaker Parameter Drift

    DEFF Research Database (Denmark)

    Pedersen, Bo Rohde

    2010-01-01

    This study tested micro loudspeaker behavior from the perspective of loudspeaker parameter drift. The main difference between traditional transducers and micro loudspeakers, apart from their size, is their suspension construction. The suspension generally is a loudspeaker's most unstable parameter......, and the study investigated temperature drift and signal dependency. There is investigated three different micro loudspeakers and compared their behavior to that of a typical bass mid-range loudspeaker unit. There is measured all linear loudspeaker parameters at different temperatures....

  18. Challenges in high accuracy surface replication for micro optics and micro fluidics manufacture

    DEFF Research Database (Denmark)

    Tosello, Guido; Hansen, Hans Nørgaard; Calaon, Matteo

    2014-01-01

    Patterning the surface of polymer components with microstructured geometries is employed in optical and microfluidic applications. Mass fabrication of polymer micro structured products is enabled by replication technologies such as injection moulding. Micro structured tools are also produced...... by replication technologies such as nickel electroplating. All replication steps are enabled by a high precision master and high reproduction fidelity to ensure that the functionalities associated with the design are transferred to the final component. Engineered surface micro structures can be either...

  19. The micro hydraulic power, a sure value

    International Nuclear Information System (INIS)

    Anon.

    2003-01-01

    The micro hydroelectricity is a proven technology which has now reached maturity. Ideal for electrification of remote sites, it also serves a complement to national electric production. A recent study carried out by the ESHA estimates the potential which still available in terms of micro hydraulic power plants at 5939 MW. The small hydro power capacity (>10 MW) installed in European union and the available potential of small hydro power are presented. (A.L.B.)

  20. The micro-step motor controller

    International Nuclear Information System (INIS)

    Hong, Kwang Pyo; Lee, Chang Hee; Moon, Myung Kook; Choi, Bung Hun; Choi, Young Hyun; Cheon, Jong Gu

    2004-11-01

    The developed micro-step motor controller can handle 4 axes stepping motor drivers simultaneously and provide high power bipolar driving mechanism with constant current mode. It can be easily controlled by manual key functions and the motor driving status is displayed by the front panel VFD. Due to the development of several kinds of communication and driving protocol, PC can operate even several micro-step motor controllers at once by multi-drop connection

  1. Concordian Economics: Beyond Micro and Macroeconomics

    OpenAIRE

    Gorga, Carmine

    2017-01-01

    In Concordian economics there is no distinction between micro and macro economics, because the economic process is the same for the individual person, the city, the nation, or the world, What changes is the scale, but not the structure of the process. When micro and macro economics are seen as one, it makes no sense to add monetary wealth to real wealth. It becomes then evident that monetary wealth is not wealth; monetary wealth is a legal representation of real wealth.

  2. Micro-battery Development using beta radioisotope

    International Nuclear Information System (INIS)

    Jung, H. K.; Cheong, Y. M.; Lee, N. H.; Choi, Y. S.; Joo, Y. S.; Lee, J. S.; Jeon, B. H.

    2007-06-01

    Nuclear battery which use the beta radiation sources emitting the low penetration radiation energy from radioisotope can be applied as the long term (more than 10 years) micro power source in MEMS and nano components. This report describes the basic concept and principles of nuclear micro-battery and its fabrication in space and military field. In particular direct conversion method is described by investigating the electron-hole generation and recombination in p-n junction of silicon betavoltaics with beta radiation

  3. Geometry and surface damage in micro electrical discharge machining of micro-holes

    Science.gov (United States)

    Ekmekci, Bülent; Sayar, Atakan; Tecelli Öpöz, Tahsin; Erden, Abdulkadir

    2009-10-01

    Geometry and subsurface damage of blind micro-holes produced by micro electrical discharge machining (micro-EDM) is investigated experimentally to explore the relational dependence with respect to pulse energy. For this purpose, micro-holes are machined with various pulse energies on plastic mold steel samples using a tungsten carbide tool electrode and a hydrocarbon-based dielectric liquid. Variations in the micro-hole geometry, micro-hole depth and over-cut in micro-hole diameter are measured. Then, unconventional etching agents are applied on the cross sections to examine micro structural alterations within the substrate. It is observed that the heat-damaged segment is composed of three distinctive layers, which have relatively high thicknesses and vary noticeably with respect to the drilling depth. Crack formation is identified on some sections of the micro-holes even by utilizing low pulse energies during machining. It is concluded that the cracking mechanism is different from cracks encountered on the surfaces when machining is performed by using the conventional EDM process. Moreover, an electrically conductive bridge between work material and debris particles is possible at the end tip during machining which leads to electric discharges between the piled segments of debris particles and the tool electrode during discharging.

  4. Geometry and surface damage in micro electrical discharge machining of micro-holes

    International Nuclear Information System (INIS)

    Ekmekci, Bülent; Sayar, Atakan; Öpöz, Tahsin Tecelli; Erden, Abdulkadir

    2009-01-01

    Geometry and subsurface damage of blind micro-holes produced by micro electrical discharge machining (micro-EDM) is investigated experimentally to explore the relational dependence with respect to pulse energy. For this purpose, micro-holes are machined with various pulse energies on plastic mold steel samples using a tungsten carbide tool electrode and a hydrocarbon-based dielectric liquid. Variations in the micro-hole geometry, micro-hole depth and over-cut in micro-hole diameter are measured. Then, unconventional etching agents are applied on the cross sections to examine micro structural alterations within the substrate. It is observed that the heat-damaged segment is composed of three distinctive layers, which have relatively high thicknesses and vary noticeably with respect to the drilling depth. Crack formation is identified on some sections of the micro-holes even by utilizing low pulse energies during machining. It is concluded that the cracking mechanism is different from cracks encountered on the surfaces when machining is performed by using the conventional EDM process. Moreover, an electrically conductive bridge between work material and debris particles is possible at the end tip during machining which leads to electric discharges between the piled segments of debris particles and the tool electrode during discharging

  5. Modulation of microRNA activity by semi-microRNAs (smiRNAs

    Directory of Open Access Journals (Sweden)

    Isabelle ePlante

    2012-06-01

    Full Text Available The ribonuclease Dicer plays a central role in the microRNA pathway by catalyzing the formation of 19 to 24-nucleotide (nt long microRNAs. Subsequently incorporated into Ago2 effector complexes, microRNAs are known to regulate messenger RNA (mRNA translation. Whether shorter RNA species derived from microRNAs exist and play a role in mRNA regulation remains unknown. Here, we report the serendipitous discovery of a 12-nt long RNA species corresponding to the 5’ region of the microRNA let-7, and tentatively termed semi-microRNA, or smiRNA. Using a smiRNA derived from the precursor of miR-223 as a model, we show that 12-nt long smiRNA species are devoid of any direct mRNA regulatory activity, as assessed in a reporter gene activity assay in transfected cultured human cells. However, smiR-223 was found to modulate the ability of the microRNA from which it derives to mediate translational repression or cleavage of reporter mRNAs. Our findings suggest that smiRNAs may be generated along the microRNA pathway and participate to the control of gene expression by regulating the activity of the related full-length mature microRNA in vivo.

  6. Laser 3D micro-manufacturing

    International Nuclear Information System (INIS)

    Piqué, Alberto; Auyeung, Raymond C Y; Kim, Heungsoo; Charipar, Nicholas A; Mathews, Scott A

    2016-01-01

    Laser-based materials processing techniques are gaining widespread use in micro-manufacturing applications. The use of laser microfabrication techniques enables the processing of micro- and nanostructures from a wide range of materials and geometries without the need for masking and etching steps commonly associated with photolithography. This review aims to describe the broad applications space covered by laser-based micro- and nanoprocessing techniques and the benefits offered by the use of lasers in micro-manufacturing processes. Given their non-lithographic nature, these processes are also referred to as laser direct-write and constitute some of the earliest demonstrations of 3D printing or additive manufacturing at the microscale. As this review will show, the use of lasers enables precise control of the various types of processing steps—from subtractive to additive—over a wide range of scales with an extensive materials palette. Overall, laser-based direct-write techniques offer multiple modes of operation including the removal (via ablative processes) and addition (via photopolymerization or printing) of most classes of materials using the same equipment in many cases. The versatility provided by these multi-function, multi-material and multi-scale laser micro-manufacturing processes cannot be matched by photolithography nor with other direct-write microfabrication techniques and offer unique opportunities for current and future 3D micro-manufacturing applications. (topical review)

  7. Micro Credit and Gender: A Critical Assessment

    Directory of Open Access Journals (Sweden)

    Özlem BALKIZ

    2015-12-01

    Full Text Available Micro credit programs, which are based on lending money on interest and encouraging savings, have been first been used in Southern countries and are now being implemented worldwide. Mainly aimed at the rural poor, particularly poor women, micro credit programs seek to ensure sustainable economic development in line with the requirements of global capitalism and to include women in the productive activities of the market. Micro credit has been made institutionalized based on three main paradigms, namely financial sustainability, poverty alleviation and women’s empowerment. In micro credit programs, where the emphasis on women’s empowerment is strong, the lack of a social gender perspective is striking. In fact, women may face patriarchal pressure and restrictions at the start in access to loans, loan usage models, participation to the productive activities in the market and during loan repayment. Thus the allegation that by way of micro credit, women will be empowered in terms of economic, social and political means in the family and society becomes questionable. This article, by problematizing women’s relationship with micro credit, will discuss social gender relationships which prevent them from making use of these programs as they wish and from achieving the results they intend

  8. MicroRNAs in right ventricular remodelling.

    Science.gov (United States)

    Batkai, Sandor; Bär, Christian; Thum, Thomas

    2017-10-01

    Right ventricular (RV) remodelling is a lesser understood process of the chronic, progressive transformation of the RV structure leading to reduced functional capacity and subsequent failure. Besides conditions concerning whole hearts, some pathology selectively affects the RV, leading to a distinct RV-specific clinical phenotype. MicroRNAs have been identified as key regulators of biological processes that drive the progression of chronic diseases. The role of microRNAs in diseases affecting the left ventricle has been studied for many years, however there is still limited information on microRNAs specific to diseases in the right ventricle. Here, we review recently described details on the expression, regulation, and function of microRNAs in the pathological remodelling of the right heart. Recently identified strategies using microRNAs as pharmacological targets or biomarkers will be highlighted. Increasing knowledge of pathogenic microRNAs will finally help improve our understanding of underlying distinct mechanisms and help utilize novel targets or biomarkers to develop treatments for patients suffering from right heart diseases. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2017. For permissions, please email: journals.permissions@oup.com.

  9. Micro-Doppler classification of riders and riderless horses

    Science.gov (United States)

    Tahmoush, David

    2014-05-01

    Micro-range Micro-Doppler can be used to isolate particular parts of the radar signature, and in this case we demonstrate the differences in the signature between a walking horse versus a walking horse with a rider. Using micro-range micro-Doppler, we can distinguish the radar returns from the rider as separate from the radar returns of the horse.

  10. Advanced Micro Turbine System (AMTS) -C200 Micro Turbine -Ultra-Low Emissions Micro Turbine

    Energy Technology Data Exchange (ETDEWEB)

    Capstone Turbine Corporation

    2007-12-31

    In September 2000 Capstone Turbine Corporation commenced work on a US Department of Energy contract to develop and improve advanced microturbines for power generation with high electrical efficiency and reduced pollutants. The Advanced MicroTurbine System (AMTS) program focused on: (1) The development and implementation of technology for a 200 kWe scale high efficiency microturbine system (2) The development and implementation of a 65 kWe microturbine which meets California Air Resources Board (CARB) emissions standards effective in 2007. Both of these objectives were achieved in the course of the AMTS program. At its conclusion prototype C200 Microturbines had been designed, assembled and successfully completed field demonstration. C65 Microturbines operating on natural, digester and landfill gas were also developed and successfully tested to demonstrate compliance with CARB 2007 Fossil Fuel Emissions Standards for NOx, CO and VOC emissions. The C65 Microturbine subsequently received approval from CARB under Executive Order DG-018 and was approved for sale in California. The United Technologies Research Center worked in parallel to successfully execute a RD&D program to demonstrate the viability of a low emissions AMS which integrated a high-performing microturbine with Organic Rankine Cycle systems. These results are documented in AMS Final Report DOE/CH/11060-1 dated March 26, 2007.

  11. C. elegans microRNAs.

    Science.gov (United States)

    Vella, Monica C; Slack, Frank J

    2005-09-21

    MicroRNAs (miRNAs) are small, non-coding regulatory RNAs found in many phyla that control such diverse events as development, metabolism, cell fate and cell death. They have also been implicated in human cancers. The C. elegans genome encodes hundreds of miRNAs, including the founding members of the miRNA family lin-4 and let-7. Despite the abundance of C. elegans miRNAs, few miRNA targets are known and little is known about the mechanism by which they function. However, C. elegans research continues to push the boundaries of discovery in this area. lin-4 and let-7 are the best understood miRNAs. They control the timing of adult cell fate determination in hypodermal cells by binding to partially complementary sites in the mRNA of key developmental regulators to repress protein expression. For example, lin-4 is predicted to bind to seven sites in the lin-14 3' untranslated region (UTR) to repress LIN-14, while let-7 is predicted to bind two let-7 complementary sites in the lin-41 3' UTR to down-regulate LIN-41. Two other miRNAs, lsy-6 and mir-273, control left-right asymmetry in neural development, and also target key developmental regulators for repression. Approximately one third of the C. elegans miRNAs are differentially expressed during development indicating a major role for miRNAs in C. elegans development. Given the remarkable conservation of developmental mechanism across phylogeny, many of the principles of miRNAs discovered in C. elegans are likely to be applicable to higher animals.

  12. STUDY & ANALYSIS OF MICRO NEEDLE MATERIAL BY ANSYS

    OpenAIRE

    Santosh Kumar Singh*, Prabhat Sinha, N.N. Singh, Nagendra Kumar

    2017-01-01

    In this research the concept of design and analysis, silicon and stainless steel based on hollow micro-needles for transdermal drug delivery(TDD) have been evaluated by Using ANSYS & computational fluid dynamic (CFD), structural. Micro fluidic analysis has performed to ensure the micro-needles design suitability for Drug delivery. The effect of axial and transverse load on single and micro-needle array has investigated with the mechanical properties of micro-needle. The analysis predicte...

  13. Selective wetting-induced micro-electrode patterning for flexible micro-supercapacitors.

    Science.gov (United States)

    Kim, Sung-Kon; Koo, Hyung-Jun; Lee, Aeri; Braun, Paul V

    2014-08-13

    Selective wetting-induced micro-electrode patterning is used to fabricate flexible micro-supercapacitors (mSCs). The resulting mSCs exhibit high performance, mechanical stability, stable cycle life, and hold great promise for facile integration into flexible devices requiring on-chip energy storage. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Elementary particles as micro-universes or micro-black holes

    International Nuclear Information System (INIS)

    Rodrigues Junior, W.A.

    1985-01-01

    The idea that elementary particles can be presented as micro-universes and/or micro-black holes (Lorentzian manifolds) is presented and the fundamental mathematical problem associated with the simplest world manifold that 'contains' both the macrocosm and the microcosmes is discussed. (Author) [pt

  15. Micro Engineering: Experiments conducted on the use of polymeric materials in micro injection moulding

    DEFF Research Database (Denmark)

    Griffiths, Christian; Tosello, Guido; Nestler, Joerg

    2008-01-01

    To advance micro injection moulding it is necessary to study systematically the factors affecting process and tooling reliability. This paper reviews the main findings of Cardiff Universities 4M and SEMOFS research in this field. In particular, the factors affecting the manufacturability of micro...

  16. Concept of subsurface micro-sensing; Chika joho no micro sensing

    Energy Technology Data Exchange (ETDEWEB)

    Niitsuma, H [Tohoku University, Sendai (Japan). Faculty of Engineering

    1997-05-27

    This paper describes concept of subsurface micro-sensing. It is intended to achieve an epoch-making development of subsurface engineerings by developing such technologies as micro measurement of well interior, micro measurement while drilling (MWD), and micro intelligent logging. These technologies are supported by development of micro sensors and micro drilling techniques using micro machine technologies. Micronizing the subsurface sensors makes mass production of sensors with equivalent performance possible, and the production cost can be reduced largely. The sensors can be embedded or used disposably, resulting in increased mobility in measurement and higher performance. Installing multiple number of sensors makes high-accuracy measurement possible, such as array measurement. The sensors can be linked easily with photo-electronics components, realizing remote measurement at low price and high accuracy. Control in micro-drilling and MWD also become possible. Such advantages may also be expected as installing the sensors on the outer side of wells in use and monitoring subsurface information during production. Expectation on them is large as a new paradigm of underground exploration and measurement. 1 fig.

  17. Fabrication of Biochips with Micro Fluidic Channels by Micro End-milling and Powder Blasting

    Directory of Open Access Journals (Sweden)

    Dong Sam Park

    2008-02-01

    Full Text Available For microfabrications of biochips with micro fluidic channels, a large number of microfabrication techniques based on silicon or glass-based Micro-Electro-Mechanical System (MEMS technologies were proposed in the last decade. In recent years, for low cost and mass production, polymer-based microfabrication techniques by microinjection molding and micro hot embossing have been proposed. These techniques, which require a proper photoresist, mask, UV light exposure, developing, and electroplating as a preprocess, are considered to have some problems. In this study, we propose a new microfabrication technology which consists of micro end-milling and powder blasting. This technique could be directly applied to fabricate the metal mold without any preprocesses. The metal mold with micro-channels is machined by micro end-milling, and then, burrs generated in the end-milling process are removed by powder blasting. From the experimental results, micro end-milling combined with powder blasting could be applied effectively for fabrication of the injection mold of biochips with micro fluidic channels.

  18. On the performance of micro injection moulding process simulations of TPE micro rings

    DEFF Research Database (Denmark)

    Baruffi, Federico; Calaon, Matteo; Tosello, Guido

    , a case study based on the micro injection moulding process of thermoplastic elastomer (TPE) micro rings (volume: 1.5 mm3, mass: 2.2 mg) for sensors application is treated. Injection moulding process simulations using Autodesk Moldflow Insight 2016® were applied with the aim of accomplishing two main...

  19. Investigation on the Acoustic Absorption of Flexible Micro-Perforated Panel with Ultra-Micro Perforations

    Science.gov (United States)

    Li, Guoxin; Tang, Xiaoning; Zhang, Xiaoxiao; Qian, Y. J.; Kong, Deyi

    2017-11-01

    Flexible micro-perforated panel has unique advantages in noise reduction due to its good flexibility compared with traditional rigid micro-perforated panel. In this paper, flexible micro-perforated panel was prepared by computer numerical control (CNC) milling machine. Three kinds of plastics including polyvinylchloride (PVC), polyethylene terephthalate (PET), and polyimide (PI) were taken as the matrix materials to prepare flexible micro-perforated panel. It has been found that flexible micro-perforated panel made of PET possessing good porosity and proper density, elastic modulus and poisson ratio exhibited the best acoustic absorption properties. The effects of various structural parameters including perforation diameter, perforation ratio, thickness and air gap have also been investigated, which would be helpful to the optimization of acoustic absorption properties.

  20. Micro injection moulding process validation for high precision manufacture of thermoplastic elastomer micro suspension rings

    DEFF Research Database (Denmark)

    Calaon, M.; Tosello, G.; Elsborg Hansen, R.

    Micro injection moulding (μIM) is one of the most suitable micro manufacturing processes for flexible mass-production of multi-material functional micro components. The technology was employed in this research used to produce thermoplastic elastomer (TPE) micro suspension rings identified...... main μIM process parameters (melt temperature, injection speed, packing pressure) using the Design of Experiment statistical technique. Measurements results demonstrated the importance of calibrating mould´s master geometries to ensure correct part production and effective quality conformance...... on the frequency in order to improve the signal quality and assure acoustic reproduction fidelity. Production quality of the TPE rings drastically influence the product functionality. In the present study, a procedure for μIM TPE micro rings production optimization has been established. The procedure entail using...

  1. Micro-Expression Recognition Using Color Spaces.

    Science.gov (United States)

    Wang, Su-Jing; Yan, Wen-Jing; Li, Xiaobai; Zhao, Guoying; Zhou, Chun-Guang; Fu, Xiaolan; Yang, Minghao; Tao, Jianhua

    2015-12-01

    Micro-expressions are brief involuntary facial expressions that reveal genuine emotions and, thus, help detect lies. Because of their many promising applications, they have attracted the attention of researchers from various fields. Recent research reveals that two perceptual color spaces (CIELab and CIELuv) provide useful information for expression recognition. This paper is an extended version of our International Conference on Pattern Recognition paper, in which we propose a novel color space model, tensor independent color space (TICS), to help recognize micro-expressions. In this paper, we further show that CIELab and CIELuv are also helpful in recognizing micro-expressions, and we indicate why these three color spaces achieve better performance. A micro-expression color video clip is treated as a fourth-order tensor, i.e., a four-dimension array. The first two dimensions are the spatial information, the third is the temporal information, and the fourth is the color information. We transform the fourth dimension from RGB into TICS, in which the color components are as independent as possible. The combination of dynamic texture and independent color components achieves a higher accuracy than does that of RGB. In addition, we define a set of regions of interests (ROIs) based on the facial action coding system and calculated the dynamic texture histograms for each ROI. Experiments are conducted on two micro-expression databases, CASME and CASME 2, and the results show that the performances for TICS, CIELab, and CIELuv are better than those for RGB or gray.

  2. Micro-optics for microfluidic analytical applications.

    Science.gov (United States)

    Yang, Hui; Gijs, Martin A M

    2018-02-19

    This critical review summarizes the developments in the integration of micro-optical elements with microfluidic platforms for facilitating detection and automation of bio-analytical applications. Micro-optical elements, made by a variety of microfabrication techniques, advantageously contribute to the performance of an analytical system, especially when the latter has microfluidic features. Indeed the easy integration of optical control and detection modules with microfluidic technology helps to bridge the gap between the macroscopic world and chip-based analysis, paving the way for automated and high-throughput applications. In our review, we start the discussion with an introduction of microfluidic systems and micro-optical components, as well as aspects of their integration. We continue with a detailed description of different microfluidic and micro-optics technologies and their applications, with an emphasis on the realization of optical waveguides and microlenses. The review continues with specific sections highlighting the advantages of integrated micro-optical components in microfluidic systems for tackling a variety of analytical problems, like cytometry, nucleic acid and protein detection, cell biology, and chemical analysis applications.

  3. Behavior of Cell on Vibrating Micro Ridges

    Directory of Open Access Journals (Sweden)

    Haruka Hino

    2015-06-01

    Full Text Available The effect of micro ridges on cells cultured at a vibrating scaffold has been studied in vitro. Several parallel lines of micro ridges have been made on a disk of transparent polydimethylsiloxane for a scaffold. To apply the vibration on the cultured cells, a piezoelectric element was attached on the outside surface of the bottom of the scaffold. The piezoelectric element was vibrated by the sinusoidal alternating voltage (Vp-p < 16 V at 1.0 MHz generated by a function generator. Four kinds of cells were used in the test: L929 (fibroblast connective tissue of C3H mouse, Hepa1-6 (mouse hepatoma, C2C12 (mouse myoblast, 3T3-L1 (mouse fat precursor cells. The cells were seeded on the micro pattern at the density of 2000 cells/cm2 in the medium containing 10% FBS (fetal bovine serum and 1% penicillin/ streptomycin. After the adhesion of cells in several hours, the cells are exposed to the ultrasonic vibration for several hours. The cells were observed with a phase contrast microscope. The experimental results show that the cells adhere, deform and migrate on the scaffold with micro patterns regardless of the ultrasonic vibration. The effects of the vibration and the micro pattern depend on the kind of cells.

  4. Role of microRNAs in sepsis.

    Science.gov (United States)

    Kingsley, S Manoj Kumar; Bhat, B Vishnu

    2017-07-01

    MicroRNAs have been found to be of high significance in the regulation of various genes and processes in the body. Sepsis is a serious clinical problem which arises due to the excessive host inflammatory response to infection. The non-specific clinical features and delayed diagnosis of sepsis has been a matter of concern for long time. MicroRNAs could enable better diagnosis of sepsis and help in the identification of the various stages of sepsis. Improved diagnosis may enable quicker and more effective treatment measures. The initial acute and transient phase of sepsis involves excessive secretion of pro-inflammatory cytokines which causes severe damage. MicroRNAs negatively regulate the toll-like receptor signaling pathway and regulate the production of inflammatory cytokines during sepsis. Likewise, microRNAs have shown to regulate the vascular barrier and endothelial function in sepsis. They are also involved in the regulation of the apoptosis, immunosuppression, and organ dysfunction in later stages of sepsis. Their importance at various levels of the pathophysiology of sepsis has been discussed along with the challenges and future perspectives. MicroRNAs could be key players in the diagnosis and staging of sepsis. Their regulation at various stages of sepsis suggests that they may have an important role in altering the outcome associated with sepsis.

  5. Heat and power from MicroGen

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    1999-10-01

    This paper reports on the design of a domestic gas-fired cogeneration system developed to replace the central heating boiler. Technical details of the MicroGen demonstration unit are given, and the use of a Linear Free Piston Stirling Engine as the prime mover, and the results of modelling studies of energy demand indicating cost savings compared to conventional boilers are discussed. The enhancement of the benefits of micro-cogeneration through use of thermal and power storage and energy demand management, and the impact of micro-cogeneration on energy use in the home are considered. The UK and European Commission's targets for increased cogeneration capacity are noted.

  6. Micro CHP: implications for energy companies

    Energy Technology Data Exchange (ETDEWEB)

    Harrison, Jeremy [EA Technology (United Kingdom); Kolin, Simon; Hestevik, Svein [Sigma Elektroteknisk A/S (Norway)

    2000-08-01

    This article explains how micro combined heat and power (CHP) technology may help UK energy businesses to maintain their customer base in the current climate of liberalisation and competition in the energy market The need for energy companies to adopt new technologies and adapt to changes in the current aggressive environment, the impact of privatisation, and the switching of energy suppliers by customers are discussed. Three potential routes to success for energy companies are identified, namely, price reductions, branding and affinity marketing, and added value services. Details are given of the implementation of schemes to encourage energy efficiency, the impact of the emissions targets set at Kyoto, the advantages of micro CHP generation, business opportunities for CHP, business threats from existing energy companies and others entering the field, and the commercial viability of micro CHP.

  7. Replication of micro and nano surface geometries

    DEFF Research Database (Denmark)

    Hansen, Hans Nørgaard; Hocken, R.J.; Tosello, Guido

    2011-01-01

    The paper describes the state-of-the-art in replication of surface texture and topography at micro and nano scale. The description includes replication of surfaces in polymers, metals and glass. Three different main technological areas enabled by surface replication processes are presented......: manufacture of net-shape micro/nano surfaces, tooling (i.e. master making), and surface quality control (metrology, inspection). Replication processes and methods as well as the metrology of surfaces to determine the degree of replication are presented and classified. Examples from various application areas...... are given including replication for surface texture measurements, surface roughness standards, manufacture of micro and nano structured functional surfaces, replicated surfaces for optical applications (e.g. optical gratings), and process chains based on combinations of repeated surface replication steps....

  8. Micro Products - Product Development and Design

    DEFF Research Database (Denmark)

    Hansen, Hans Nørgaard

    2003-01-01

    Innovation within the field of micro and nano technology is to a great extent characterized by cross-disciplinary skills. The traditional disciplines like e.g. physics, biology, medicine and engineering are united in a common development process that can only take place in the presence of multi......-disciplinary competences. One example is sensors for chemical analysis of fluids, where chemistry, biology and flow mechanics all influence the design of the product and thereby the industrial fabrication of the product [1]. On the technological side the development has moved very fast, primarily driven by the need...... of the electronics industry to create still smaller chips with still larger capacity. Therefore the manufacturing technologies connected with micro/nano products in silicon are relatively highly developed compared to the technologies used for manufacturing micro products in metals, polymers and ceramics. For all...

  9. MicroRNAs, epigenetics and disease

    DEFF Research Database (Denmark)

    Silahtaroglu, Asli; Stenvang, Jan

    2010-01-01

    Epigenetics is defined as the heritable chances that affect gene expression without changing the DNA sequence. Epigenetic regulation of gene expression can be through different mechanisms such as DNA methylation, histone modifications and nucleosome positioning. MicroRNAs are short RNA molecules...... which do not code for a protein but have a role in post-transcriptional silencing of multiple target genes by binding to their 3' UTRs (untranslated regions). Both epigenetic mechanisms, such as DNA methylation and histone modifications, and the microRNAs are crucial for normal differentiation...... diseases. In the present chapter we will mainly focus on microRNAs and methylation and their implications in human disease, mainly in cancer....

  10. Laser based micro forming and assembly.

    Energy Technology Data Exchange (ETDEWEB)

    MacCallum, Danny O' Neill; Wong, Chung-Nin Channy; Knorovsky, Gerald Albert; Steyskal, Michele D.; Lehecka, Tom (Pennsylvania State University, Freeport, PA); Scherzinger, William Mark; Palmer, Jeremy Andrew

    2006-11-01

    It has been shown that thermal energy imparted to a metallic substrate by laser heating induces a transient temperature gradient through the thickness of the sample. In favorable conditions of laser fluence and absorptivity, the resulting inhomogeneous thermal strain leads to a measurable permanent deflection. This project established parameters for laser micro forming of thin materials that are relevant to MESA generation weapon system components and confirmed methods for producing micrometer displacements with repeatable bend direction and magnitude. Precise micro forming vectors were realized through computational finite element analysis (FEA) of laser-induced transient heating that indicated the optimal combination of laser heat input relative to the material being heated and its thermal mass. Precise laser micro forming was demonstrated in two practical manufacturing operations of importance to the DOE complex: micrometer gap adjustments of precious metal alloy contacts and forming of meso scale cones.

  11. The Aarhus Ion Micro-Trap Project

    DEFF Research Database (Denmark)

    Miroshnychenko, Yevhen; Nielsen, Otto; Poulsen, Gregers

    As part of our involvement in the EU MICROTRAP project, we have designed, manufactured and assembled a micro-scale ion trap with integrated optical fibers. These prealigned fibers will allow delivering cooling laser light to single ions. Therefore, such a trap will not require any direct optical...... and installed in an ultra high vacuum chamber, which includes an ablation oven for all-optical loading of the trap [2]. The next steps on the project are to demonstrate the operation of the micro-trap and the cooling of ions using fiber delivered light. [1] D. Grant, Development of Micro-Scale Ion traps, Master...... Thesis (2008). [2] R.J. Hendricks, D.M. Grant, P.F. Herskind, A. Dantan and M. Drewsen, An all-optical ion-loading technique for scalable microtrap architectures, Applied Physics B, 88, 507 (2007)....

  12. Optical nano and micro actuator technology

    CERN Document Server

    Knopf, George K

    2012-01-01

    In Optical Nano and Micro Actuator Technology, leading engineers, material scientists, chemists, physicists, laser scientists, and manufacturing specialists offer an in-depth, wide-ranging look at the fundamental and unique characteristics of light-driven optical actuators. They discuss how light can initiate physical movement and control a variety of mechanisms that perform mechanical work at the micro- and nanoscale. The book begins with the scientific background necessary for understanding light-driven systems, discussing the nature of light and the interaction between light and NEMS/MEMS d

  13. Micro- and nanotechnology in cardiovascular tissue engineering

    International Nuclear Information System (INIS)

    Zhang Boyang; Xiao Yun; Hsieh, Anne; Thavandiran, Nimalan; Radisic, Milica

    2011-01-01

    While in nature the formation of complex tissues is gradually shaped by the long journey of development, in tissue engineering constructing complex tissues relies heavily on our ability to directly manipulate and control the micro-cellular environment in vitro. Not surprisingly, advancements in both microfabrication and nanofabrication have powered the field of tissue engineering in many aspects. Focusing on cardiac tissue engineering, this paper highlights the applications of fabrication techniques in various aspects of tissue engineering research: (1) cell responses to micro- and nanopatterned topographical cues, (2) cell responses to patterned biochemical cues, (3) controlled 3D scaffolds, (4) patterned tissue vascularization and (5) electromechanical regulation of tissue assembly and function.

  14. Rectenna session: Micro aspects. [energy conversion

    Science.gov (United States)

    Gutmann, R. J.

    1980-01-01

    Two micro aspects of the rectenna design are addressed: evaluation of the degradation in net rectenna RF to DC conversion efficiency due to power density variations across the rectenna (power combining analysis) and design of Yagi-Uda receiving elements to reduce rectenna cost by decreasing the number of conversion circuits (directional receiving elements). The first of these micro aspects involves resolving a fundamental question of efficiency potential with a rectenna, while the second involves a design modification with a large potential cost saving.

  15. Injection moulding for macro and micro products

    DEFF Research Database (Denmark)

    Islam, Mohammad Aminul

    used for macro products but with the ages it is going deep into the micro areas having machine and process improvements. Extensive research work on injection moulding is going on all over the world. New ideas are flowing into the machines, materials and processes. The technology has made significant......The purpose of the literature survey is to investigate the injection moulding technology in the macro and micro areas from the basic to the state-of-the-art recent technology. Injection moulding is a versatile production process for the manufacturing of plastic parts and the process is extensively...

  16. Micro- and nanoscale phenomena in tribology

    CERN Document Server

    Chung, Yip-Wah

    2011-01-01

    Drawn from presentations at a recent National Science Foundation Summer Institute on Nanomechanics, Nanomaterials, and Micro/Nanomanufacturing, Micro- and Nanoscale Phenomena in Tribology explores the convergence of the multiple science and engineering disciplines involved in tribology and the connection from the macro to nano world. Written by specialists from computation, materials science, mechanical engineering, surface physics, and chemistry, each chapter provides up-to-date coverage of both basic and advanced topics and includes extensive references for further study.After discussing the

  17. MicroRNAs in Prostate Cancer

    Science.gov (United States)

    2008-11-01

    lymphoma. Genes Chromosom. Cancer 39:167–69 131. O’Connell RM, Taganov KD, Boldin MP, Cheng G, Baltimore D. 2007. MicroRNA-155 is induced during the...carcinoma. J. Virol. 81:1033–36 155. Xi Y, Nakajima G, Gavin E, Morris CG, Kudo K, et al. 2007. Systematic analysis of microRNA expression of RNA extracted ...diversity. miRNAs were extracted from the unique sequences by searching against miRNA database (miRbase release 10.0; http://microrna.sanger.ac.uk

  18. Demoulding force in micro-injection moulding

    DEFF Research Database (Denmark)

    Griffiths, C.A.; Dimov, S.S.; Scholz, S.

    2012-01-01

    The paper reports an experimental study that investigates part demoulding behavior in micro injection moulding (MIM) with a focus on the effects of pressure (P) and temperature (T) on the demoulding forces. Demoulding of a microfluidics part is conducted and the four processing parameters of melt...... temperature (Tb), mould temperature (Tm), holding pressure (Ph) and injection speed (Vi) are analysed. The result using different combinations of process parameters were used to identify the best processing conditions in regards to demoulding forces when moulding micro parts....

  19. Micro-Scale Avionics Thermal Management

    Science.gov (United States)

    Moran, Matthew E.

    2001-01-01

    Trends in the thermal management of avionics and commercial ground-based microelectronics are converging, and facing the same dilemma: a shortfall in technology to meet near-term maximum junction temperature and package power projections. Micro-scale devices hold the key to significant advances in thermal management, particularly micro-refrigerators/coolers that can drive cooling temperatures below ambient. A microelectromechanical system (MEMS) Stirling cooler is currently under development at the NASA Glenn Research Center to meet this challenge with predicted efficiencies that are an order of magnitude better than current and future thermoelectric coolers.

  20. Towards the first generation micro bulk forming system

    DEFF Research Database (Denmark)

    Arentoft, Mogens; Eriksen, Rasmus Solmer; Hansen, Hans Nørgaard

    2011-01-01

    . This work describes a number of prototype system units, which collectively form a desktop sized micro forming production system. The system includes a billet preparation module, an integrated transfer system, a temperature controlled forming tool, including process simulation, and a dedicated micro forming......The industrial demand for micro mechanical components has surged in the later years with the constant introduction of more integrated products. The micro bulk forming process holds a promising pledge of delivering high quality micro mechanical components at low cost and high production rates...... press. The system is demonstrated on an advanced micro forming case where a dental component is formed in medical grade Titanium....

  1. Modeling and simulation for micro DC motor based on simulink

    Science.gov (United States)

    Shen, Hanxin; Lei, Qiao; Chen, Wenxiang

    2017-09-01

    The micro DC motor has a large market demand but there is a lack of theoretical research for it. Through detailed analysis of the commutation process of micro DC motor commutator, based on micro DC motor electromagnetic torque equation and mechanical torque equation, with the help of Simulink toolkit, a triangle connection micro DC motor simulation model is established. By using the model, a sample micro DC motor are simulated, and an experimental measurements has been carried on the sample micro DC motor. It is found that the simulation results are consistent with theoretical analysis and experimental results.

  2. Gas Mixtures for Welding with Micro-Jet Cooling

    Directory of Open Access Journals (Sweden)

    Węgrzyn T.

    2015-04-01

    Full Text Available Welding with micro-jet cooling after was tested only for MIG and MAG processes. For micro-jet gases was tested only argon, helium and nitrogen. A paper presents a piece of information about gas mixtures for micro-jet cooling after in welding. There are put down information about gas mixtures that could be chosen both for MAG welding and for micro-jet process. There were given main information about influence of various micro-jet gas mixtures on metallographic structure of steel welds. Mechanical properties of weld was presented in terms of various gas mixtures selection for micro-jet cooling.

  3. System program for MICRO-CAMAC terminal system

    International Nuclear Information System (INIS)

    Sasajima, Yoji; Yamada, Takayuki; Yagi, Hideyuki; Ishiguro, Misako

    1979-08-01

    A JAERI on-line network system was developed and exists for on-line data processing of nuclear instrumentation. As terminal systems for the network system, the one with a Micro -8 micro-computer is used. By modifying the control program for Micro-8 terminal system, a system program has been developed for a MICRO-CAMAC terminal system, which is controlled by a micro-computer framed within the CAMAC Crate Controller. In this report are described software specifications of the MICRO -CAMAC terminal system and its operation method. (author)

  4. Fabrication of a Flexible Micro CO Sensor for Micro Reformer Applications

    Directory of Open Access Journals (Sweden)

    Yi-Man Lo

    2010-11-01

    Full Text Available Integration of a reformer and a proton exchange membrane fuel cell (PEMFC is problematic due to the presence in the gas from the reforming process of a slight amount of carbon monoxide. Carbon monoxide poisons the catalyst of the proton exchange membrane fuel cell subsequently degrading the fuel cell performance, and necessitating the sublimation of the reaction gas before supplying to fuel cells. Based on the use of micro-electro-mechanical systems (MEMS technology to manufacture flexible micro CO sensors, this study elucidates the relation between a micro CO sensor and different SnO2 thin film thicknesses. Experimental results indicate that the sensitivity increases at temperatures ranging from 100–300 °C. Additionally, the best sensitivity is obtained at a specific temperature. For instance, the best sensitivity of SnO2 thin film thickness of 100 nm at 300 °C is 59.3%. Moreover, a flexible micro CO sensor is embedded into a micro reformer to determine the CO concentration in each part of a micro reformer in the future, demonstrating the inner reaction of a micro reformer in depth and immediate detection.

  5. A novel integrated multifunction micro-sensor for three-dimensional micro-force measurements.

    Science.gov (United States)

    Wang, Weizhong; Zhao, Yulong; Qin, Yafei

    2012-01-01

    An integrated multifunction micro-sensor for three-dimensional micro-force precision measurement under different pressure and temperature conditions is introduced in this paper. The integrated sensor consists of three kinds of sensors: a three-dimensional micro-force sensor, an absolute pressure sensor and a temperature sensor. The integrated multifunction micro-sensor is fabricated on silicon wafers by micromachining technology. Different doping doses of boron ion, placement and structure of resistors are tested for the force sensor, pressure sensor and temperature sensor to minimize the cross interference and optimize the properties. A glass optical fiber, with a ladder structure and sharp tip etched by buffer oxide etch solution, is glued on the micro-force sensor chip as the tactile probe. Experimental results show that the minimum force that can be detected by the force sensor is 300 nN; the lateral sensitivity of the force sensor is 0.4582 mV/μN; the probe length is linearly proportional to sensitivity of the micro-force sensor in lateral; the sensitivity of the pressure sensor is 0.11 mv/KPa; the sensitivity of the temperature sensor is 5.836 × 10(-3) KΩ/°C. Thus it is a cost-effective method to fabricate integrated multifunction micro-sensors with different measurement ranges that could be used in many fields.

  6. A Novel Integrated Multifunction Micro-Sensor for Three-Dimensional Micro-Force Measurements

    Directory of Open Access Journals (Sweden)

    Yafei Qin

    2012-03-01

    Full Text Available An integrated multifunction micro-sensor for three-dimensional micro-force precision measurement under different pressure and temperature conditions is introduced in this paper. The integrated sensor consists of three kinds of sensors: a three-dimensional micro-force sensor, an absolute pressure sensor and a temperature sensor. The integrated multifunction micro-sensor is fabricated on silicon wafers by micromachining technology. Different doping doses of boron ion, placement and structure of resistors are tested for the force sensor, pressure sensor and temperature sensor to minimize the cross interference and optimize the properties. A glass optical fiber, with a ladder structure and sharp tip etched by buffer oxide etch solution, is glued on the micro-force sensor chip as the tactile probe. Experimental results show that the minimum force that can be detected by the force sensor is 300 nN; the lateral sensitivity of the force sensor is 0.4582 mV/μN; the probe length is linearly proportional to sensitivity of the micro-force sensor in lateral; the sensitivity of the pressure sensor is 0.11 mv/KPa; the sensitivity of the temperature sensor is 5.836 × 10−3 KΩ/°C. Thus it is a cost-effective method to fabricate integrated multifunction micro-sensors with different measurement ranges that could be used in many fields.

  7. High accuracy and precision micro injection moulding of thermoplastic elastomers micro ring production

    DEFF Research Database (Denmark)

    Calaon, Matteo; Tosello, Guido; Elsborg, René

    2016-01-01

    The mass-replication nature of the process calls for fast monitoring of process parameters and product geometrical characteristics. In this direction, the present study addresses the possibility to develop a micro manufacturing platform for micro assembly injection moulding with real-time process....../product monitoring and metrology. The study represent a new concept yet to be developed with great potential for high precision mass-manufacturing of highly functional 3D multi-material (i.e. including metal/soft polymer) micro components. The activities related to HINMICO project objectives proves the importance...

  8. Micro-masters of glioblastoma biology and therapy: increasingly recognized roles for microRNAs.

    Science.gov (United States)

    Floyd, Desiree; Purow, Benjamin

    2014-05-01

    MicroRNAs are small noncoding RNAs encoded in eukaryotic genomes that have been found to play critical roles in most biological processes, including cancer. This is true for glioblastoma, the most common and lethal primary brain tumor, for which microRNAs have been shown to strongly influence cell viability, stem cell characteristics, invasiveness, angiogenesis, metabolism, and immune evasion. Developing microRNAs as prognostic markers or as therapeutic agents is showing increasing promise and has potential to reach the clinic in the next several years. This succinct review summarizes current progress and future directions in this exciting and steadily expanding field.

  9. Morphology of Major Stone Types, As Shown by Micro Computed Tomography (micro CT)

    International Nuclear Information System (INIS)

    Jackson, Molly E.; Beuschel, Christian A.; McAteer, James A.; Williams, James C.

    2008-01-01

    Micro CT offers the possibility of providing a non-destructive method of stone analysis that allows visualization of 100% of the stone's volume. For the present study, micro CT analysis was completed on stones of known composition with isotropic voxel sizes of either 7 or 9.1 μm. Each mineral type was distinctive, either by x-ray attenuation values or by morphology. Minor components, such as the presence of apatite in oxalate stones, were easily seen. The analysis of stones by micro CT opens up the possibility of exploring the stone as an encapsulated history of the patient's disease, showing changes in mineral deposition with time.

  10. Design and fabrication of self-powered micro-harvesters rotating and vibrated micro-power systems

    CERN Document Server

    Pan, C T; Lin, Liwei; Chen, Ying-Chung

    2013-01-01

    Presents the latest methods for designing and fabricating self-powered micro-generators and energy harvester systems Design and Fabrication of Self-Powered Micro-Harvesters introduces the latest trends of self-powered generators and energy harvester systems, including the design, analysis and fabrication of micro power systems. Presented in four distinct parts, the authors explore the design and fabrication of: vibration-induced electromagnetic micro-generators; rotary electromagnetic micro-generators; flexible piezo-micro-generator with various widths; and PVDF electrospunpiezo-energy with

  11. Micro-thermal analysis of polyester coatings

    NARCIS (Netherlands)

    Fischer, H.R.

    2010-01-01

    The application and suitability of micro-thermal analysis to detect changes in the chemical and physical properties of coating due to ageing and especially photo-degradation is demonstrated using a model polyester coating based on neopentyl glycol isophthalic acid. The changes in chemical structure

  12. Hollow micro string based calorimeter device

    DEFF Research Database (Denmark)

    2014-01-01

    positions so as to form a free released double clamped string in-between said two longitudinally distanced positions said micro-channel string comprising a microfluidic channel having a closed cross section and extending in the longitudinal direction of the hollow string, acoustical means adapted...

  13. Micro Econometric Modelling of Household Energy Use

    DEFF Research Database (Denmark)

    Leth-Petersen, Søren

    2002-01-01

    Presents a micro econometric analysis of household electricity and natural gas demand for Danish households observed in 1996. Dependence between demand for gas and demand for electricity; Separability of demand for gas from demand for electricity; Relation between energy consumption and the age...

  14. MicroRNA mimicry blocks pulmonary fibrosis

    NARCIS (Netherlands)

    Montgomery, Rusty L; Yu, Guoying; Latimer, Paul A; Stack, Christianna; Robinson, Kathryn; Dalby, Christina M; Kaminski, Naftali; van Rooij, Eva

    2014-01-01

    Over the last decade, great enthusiasm has evolved for microRNA (miRNA) therapeutics. Part of the excitement stems from the fact that a miRNA often regulates numerous related mRNAs. As such, modulation of a single miRNA allows for parallel regulation of multiple genes involved in a particular

  15. Micro-Macro Paradoxes of Entrepreneurship

    DEFF Research Database (Denmark)

    Søgaard, Villy

    2008-01-01

    Artiklen tager afsæt i det såkaldte micro-macro paradox fra Aids-Efficiency litteraturen og argumenterer for, at en tilsvarende problemstilling bør inddrages i vurderingen af f.eks. de beskæftigelsesmæssige konsekvenser af entrepreneuriel virksomhed. Den påviser også i en gennemgang af litteratur...

  16. Thermal property testing technique on micro specimen

    International Nuclear Information System (INIS)

    Baba, Tetsuya; Kishimoto, Isao; Taketoshi, Naoyuki

    2000-01-01

    This study aims at establishment of further development on some testing techniques on the nuclear advanced basic research accumulated by the National Research Laboratory of Metrology for ten years. For this purpose, a technology to test heat diffusion ratio and specific heat capacity of less than 3 mm in diameter and 1 mm in thickness of micro specimen and technology to test heat diffusion ratio at micro area of less than 1 mm in area along cross section of less than 10 mm in diameter of column specimen were developed to contribute to common basic technology supporting the nuclear power field. As a result, as an element technology to test heat diffusion ratio and specific heat capacity of the micro specimen, a specimen holding technique stably to hold a micro specimen with 3 mm in diameter could be developed. And, for testing the specific heat capacity by using the laser flush differential calorimetry, a technique to hold two specimen of 5 mm in diameter at their proximities was also developed. In addition, by promoting development of thermal property data base capable of storing thermal property data obtained in this study and with excellent workability in this 1998 fiscal year a data in/out-put program with graphical user interface could be prepared. (G.K.)

  17. Recent advances in micro-vibration isolation

    Science.gov (United States)

    Liu, Chunchuan; Jing, Xingjian; Daley, Steve; Li, Fengming

    2015-05-01

    Micro-vibration caused by disturbance sources onboard spacecraft can severely degrade the working environment of sensitive payloads. Some notable vibration control methods have been developed particularly for the suppression or isolation of micro-vibration over recent decades. Usually, passive isolation techniques are deployed in aerospace engineering. Active isolators, however, are often proposed to deal with the low frequency vibration that is common in spacecraft. Active/passive hybrid isolation has also been effectively used in some spacecraft structures for a number of years. In semi-active isolation systems, the inherent structural performance can be adjusted to deal with variation in the aerospace environment. This latter approach is potentially one of the most practical isolation techniques for micro-vibration isolation tasks. Some emerging advanced vibration isolation methods that exploit the benefits of nonlinearity have also been reported in the literature. This represents an interesting and highly promising approach for solving some challenging problems in the area. This paper serves as a state-of-the-art review of the vibration isolation theory and/or methods which were developed, mainly over the last decade, specifically for or potentially could be used for, micro-vibration control.

  18. Power Talk in DC Micro Grids

    DEFF Research Database (Denmark)

    Angjelichinoski, Marko; Stefanovic, Cedomir; Popovski, Petar

    2015-01-01

    Power talk is a novel concept for communication among units in a Micro Grid (MG), where information is sent by using power electronics as modems and the common bus of the MG as a communication medium. The technique is implemented by modifying the droop control parameters from the primary control...

  19. Micro and Small Enterprises Incubator - Phase III

    International Development Research Centre (IDRC) Digital Library (Canada)

    The goals of the Mozambique Information and Communication Technology Micro and Small Enterprises Incubator (MICTI Incubator) are twofold: to identify sustainable opportunities for technology-based businesses in priority development areas; and to test the assumption that technology-based businesses can mentor the ...

  20. Introducing Micro-finance in Sweden

    DEFF Research Database (Denmark)

    Barinaga, Ester

    2013-01-01

    The case describes the first year of efforts to introduce microfinance as a tool to work with vulnerable groups in Sweden, more particularly ex-convicts, former drug-addicts and longterm unemployed women of immigrant background. The teaching objective is to discuss whether micro-finance can be seen...

  1. OSH management in small and micro enterprises

    NARCIS (Netherlands)

    Zwetsloot, G.I.J.M.

    2014-01-01

    Small and medium-sized enterprises (SMEs) are widely acknowledged as the backbone of the European economy. According to EUROSTAT statistics [1], [2], 29.6% of the EU employees work in micro enterprises (<10 employees), while 20.6 % are employed in small firms (<50 employees). Indeed, half of the

  2. Micro-channel plates and vacuum detectors

    Energy Technology Data Exchange (ETDEWEB)

    Gys, T., E-mail: Thierry.Gys@cern.ch

    2015-07-01

    A micro-channel plate is an array of miniature electron multipliers that are each acting as a continuous dynode chain. The compact channel structure results in high spatial and time resolutions and robustness to magnetic fields. Micro-channel plates have been originally developed for night vision applications and integrated as an amplification element in image intensifiers. These devices show single-photon sensitivity with very low noise and have been used as such for scintillating fiber tracker readout in high-energy physics experiments. Given their very short transit time spread, micro-channel plate photomultiplier tubes are also being used in time-of-flight and particle identification detectors. The present paper will cover the history of the micro-channel plate development, basic features, and some of their applications. Emphasis will be put on various new manufacturing processes that have been developed over the last few years, and that result in a significant improvement in terms of efficiency, noise, and lifetime performance.

  3. The Micro-Category Account of Analogy

    Science.gov (United States)

    Green, Adam E.; Fugelsang, Jonathan A.; Kraemer, David J. M.; Dunbar, Kevin N.

    2008-01-01

    Here, we investigate how activation of mental representations of categories during analogical reasoning influences subsequent cognitive processing. Specifically, we present and test the central predictions of the "Micro-Category" account of analogy. This account emphasizes the role of categories in aligning terms for analogical mapping. In a…

  4. Mozambique Information and Communication Technology : Micro ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    The goals of the Mozambique Information and Communication Technology Micro and Small Enterprises Incubator (MICTI Incubator) are twofold: to identify sustainable opportunities for technology-based businesses in priority development areas; and to test the assumption that technology-based businesses can mentor the ...

  5. MicroRNAs in skin tissue engineering.

    Science.gov (United States)

    Miller, Kyle J; Brown, David A; Ibrahim, Mohamed M; Ramchal, Talisha D; Levinson, Howard

    2015-07-01

    35.2 million annual cases in the U.S. require clinical intervention for major skin loss. To meet this demand, the field of skin tissue engineering has grown rapidly over the past 40 years. Traditionally, skin tissue engineering relies on the "cell-scaffold-signal" approach, whereby isolated cells are formulated into a three-dimensional substrate matrix, or scaffold, and exposed to the proper molecular, physical, and/or electrical signals to encourage growth and differentiation. However, clinically available bioengineered skin equivalents (BSEs) suffer from a number of drawbacks, including time required to generate autologous BSEs, poor allogeneic BSE survival, and physical limitations such as mass transfer issues. Additionally, different types of skin wounds require different BSE designs. MicroRNA has recently emerged as a new and exciting field of RNA interference that can overcome the barriers of BSE design. MicroRNA can regulate cellular behavior, change the bioactive milieu of the skin, and be delivered to skin tissue in a number of ways. While it is still in its infancy, the use of microRNAs in skin tissue engineering offers the opportunity to both enhance and expand a field for which there is still a vast unmet clinical need. Here we give a review of skin tissue engineering, focusing on the important cellular processes, bioactive mediators, and scaffolds. We further discuss potential microRNA targets for each individual component, and we conclude with possible future applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Meissner-levitated micro-systems

    Energy Technology Data Exchange (ETDEWEB)

    Coombs, T A; Samad, I; Hong, Z; Eves, D; Rastogi, A [Cambridge University Engineering Department, Trumpington Street, Cambridge, CB2 PZ (United Kingdom)

    2006-06-01

    Advanced silicon processing techniques developed for the Very Large Scale Integration (VLSI) industry have been exploited in recent years to enable the production of micro-fabricated moving mechanical systems known as Micro Electro Mechanical Systems (MEMS). These devices offer advantages in terms of cost, scalability and robustness over their preceding equivalents. Cambridge University have worked for many years on the investigation of high temperature superconductors (HTS) in flywheel energy storage applications. This experience is now being used to research into superconducting Micro-Bearings for MEMS, whereby circular permanent magnet arrays are levitated and spun above a superconductor to produce bearings suitable for motors and other micron scale devices. The novelty in the device lies in the fact that the rotor is levitated into position by Meissner flux exclusion, whilst stability is provided by flux pinned within the body of the superconductor. This work includes: the investigation of the properties of various magnetic materials, their fabrication processes and their suitability for MEMS; finite element analysis to analyse the interaction between the magnetic materials and YBCO to determine the stiffness and height of levitation. Finally a micro-motor with the above principles is currently being fabricated within the group.

  7. 3D Programmable Micro Self Assembly

    National Research Council Canada - National Science Library

    Bohringer, Karl F; Parviz, Babak A; Klavins, Eric

    2005-01-01

    .... We have developed a "self assembly tool box" consisting of a range of methods for micro-scale self-assembly in 2D and 3D We have shown physical demonstrations of simple 3D self-assemblies which lead...

  8. MEMS Micro-Valve for Space Applications

    Science.gov (United States)

    Chakraborty, I.; Tang, W. C.; Bame, D. P.; Tang, T. K.

    1998-01-01

    We report on the development of a Micro-ElectroMechanical Systems (MEMS) valve that is designed to meet the rigorous performance requirements for a variety of space applications, such as micropropulsion, in-situ chemical analysis of other planets, or micro-fluidics experiments in micro-gravity. These systems often require very small yet reliable silicon valves with extremely low leak rates and long shelf lives. Also, they must survive the perils of space travel, which include unstoppable radiation, monumental shock and vibration forces, as well as extreme variations in temperature. Currently, no commercial MEMS valve meets these requirements. We at JPL are developing a piezoelectric MEMS valve that attempts to address the unique problem of space. We begin with proven configurations that may seem familiar. However, we have implemented some major design innovations that should produce a superior valve. The JPL micro-valve is expected to have an extremely low leak rate, limited susceptibility to particulates, vibration or radiation, as well as a wide operational temperature range.

  9. Micro-PIXE for single cell analysis

    International Nuclear Information System (INIS)

    Ortega, Richard

    2012-01-01

    The knowledge of the intracellular distribution of biological relevant metals is important to understand their mechanisms of action in cells, either for physiological, toxicological or pathological processes. However, the direct detection of trace metals in single cells is a challenging task that requires sophisticated analytical developments. The combination of micro-PIXE with RBS and STIM (Scanning Transmission Ion Microscopy) allows the quantitative determination of trace metal content within sub-cellular compartments. The application of STIM analysis provides high spatial resolution imaging (< 200 nm) and excellent mass sensitivity (< 0.1 ng). Application of the STIM-PIXE-RBS methodology is absolutely needed when organic mass loss appears during PIXE-RBS irradiation. This combination of STIM-PIXE-RBS provides fully quantitative determination of trace element content, expressed in μg/g, which is a quite unique capability for micro-PIXE compared to other micro-analytical methods such as the electron and synchrotron x-ray fluorescence. Examples of micro-PIXE studies for sub-cellular imaging of trace elements in various fields of interest will be presented: in patho-physiology of trace elements involved in neurodegenerative diseases such as Parkinson's disease, and in toxicology of metals such as cobalt. (author)

  10. MICRO AUTO GASIFICATION SYSTEM: EMISSIONS CHARACTERIZATION

    Science.gov (United States)

    A compact, CONEX-housed waste to energy unit, Micro Auto Gasification System (MAGS), was characterized for air emissions from burning of military waste types. The MAGS unit is a dual chamber gasifier with a secondary diesel-fired combustor. Eight tests were conducted with multipl...

  11. Characterization of cellulose nanofibrillation by micro grinding

    Science.gov (United States)

    Sandeep S. Nair; J.Y. Zhu; Yulin Deng; Arthur J. Ragauskas

    2014-01-01

    A fundamental understanding of the morphological development of cellulose fibers during fibrillation using micro grinder is very essential to develop effective strategies for process improvement and to reduce energy consumption. We demonstrated some simple measures for characterizing cellulose fibers fibrillated at different fibrillation times through the grinder. The...

  12. Targeting of microRNAs for therapeutics

    DEFF Research Database (Denmark)

    Stenvang, Jan; Lindow, Morten; Kauppinen, Sakari

    2008-01-01

    miRNAs (microRNAs) comprise a class of small endogenous non-coding RNAs that post-transcriptionally repress gene expression by base-pairing with their target mRNAs. Recent evidence has shown that miRNAs play important roles in a wide variety of human diseases, such as viral infections, cancer...

  13. Mechanics over micro and nano scales

    CERN Document Server

    Chakraborty, Suman

    2011-01-01

    Discusses the fundaments of mechanics over micro and nano scales in a level accessible to multi-disciplinary researchers, with a balance of mathematical details and physical principles Covers life sciences and chemistry for use in emerging applications related to mechanics over small scales Demonstrates the explicit interconnection between various scale issues and the mechanics of miniaturized systems

  14. MicroRNAs, Regulatory Networks, and Comorbidities

    DEFF Research Database (Denmark)

    Russo, Francesco; Belling, Kirstine; Jensen, Anders Boeck

    2017-01-01

    MicroRNAs (miRNAs) are small noncoding RNAs involved in the posttranscriptional regulation of messenger RNAs (mRNAs). Each miRNA targets a specific set of mRNAs. Upon binding the miRNA inhibits mRNA translation or facilitate mRNA degradation. miRNAs are frequently deregulated in several pathologies...

  15. Microfluidics microFACS for Life Detection

    Science.gov (United States)

    Platt, Donald W.; Hoover, Richard B.

    2010-01-01

    A prototype micro-scale Fluorescent Activated Cell Sorter (microFACS) for life detection has been built and is undergoing testing. A functional miniature microfluidics instrument with the ability to remotely distinguish live or dead bacterial cells from abiotic particulates in ice or permafrost of icy bodies of the solar system would be of fundamental value to NASA. The use of molecular probes to obtain the bio-signature of living or dead cells could answer the most fundamental question of Astrobiology: Does life exist beyond Earth? The live-dead fluorescent stains to be used in the microFACS instrument function only with biological cell walls. The detection of the cell membranes of living or dead bacteria (unlike PAH's and many other Biomarkers) would provide convincing evidence of present or past life. This miniature device rapidly examine large numbers of particulates from a polar ice or permafrost sample and distinguish living from dead bacteria cells and biological cells from mineral grains and abiotic particulates and sort the cells and particulates based on a staining system. Any sample found to exhibit fluorescence consistent with living cells could then be used in conjunction with a chiral labeled release experiment or video microscopy system to seek addition evidence for cellular metabolism or motility. Results of preliminary testing and calibration of the microFACS prototype instrument system with pure cultures and enrichment assemblages of microbial extremophiles will be reported.

  16. The Fail-Safe Micro Research Paper.

    Science.gov (United States)

    Saunders, Mary Anne

    A key element in a research paper writing assignment modified for students of English as a second language to assure their success is teacher control over most of the process. A chronological plan for action for the micro research project includes these steps: creating an awareness of current events and controversial issues, practicing necessary…

  17. AFM plough YBCO micro bridges: substrate effects

    CSIR Research Space (South Africa)

    Elkaseh, A

    2009-04-01

    Full Text Available AFM nanolithography was used as a novel cutting technique to define micro-size YBCO superconducting constrictions. Researchers studied the substrate effects on MgO and STO substrates and showed that the observed Shapiro steps from the bridges on STO...

  18. SU-8 micro Coriolis mass flow sensor

    NARCIS (Netherlands)

    Monge, Rosa; Groenesteijn, Jarno; Alveringh, Dennis; Wiegerink, Remco J.; Lötters, Joost Conrad; Fernandez, Luis J.

    2017-01-01

    Abstract This work presents the modelling, design, fabrication and test of the first micro Coriolis mass flow sensor fully fabricated in SU-8 by photolithography processes. The sensor consists of a channel with rectangular cross-section with inner opening of 100 μm × 100 μm and is actuated at

  19. Summary of measurements with MicroVent

    DEFF Research Database (Denmark)

    Dreau, Jerome Le; Heiselberg, Per Kvols; Jensen, Rasmus Lund

    This summary presents the main results when MicroVent is used in the cooling case, without heat recovery. Experiments have thus been performed with relatively low inlet air temperature (below 15°C). Different solutions have been compared to decrease the risk of draught in the occupied zone: ‐ usi...

  20. Special Application Thermoelectric Micro Isotope Power Sources

    International Nuclear Information System (INIS)

    Heshmatpour, Ben; Lieberman, Al; Khayat, Mo; Leanna, Andrew; Dobry, Ted

    2008-01-01

    Promising design concepts for milliwatt (mW) size micro isotope power sources (MIPS) are being sought for use in various space and terrestrial applications, including a multitude of future NASA scientific missions and a range of military applications. To date, the radioisotope power sources (RPS) used on various space and terrestrial programs have provided power levels ranging from one-half to several hundred watts. In recent years, the increased use of smaller spacecraft and planned new scientific space missions by NASA, special terrestrial and military applications suggest the need for lower power, including mW level, radioisotope power sources. These power sources have the potential to enable such applications as long-lived meteorological or seismological stations distributed across planetary surfaces, surface probes, deep space micro-spacecraft and sub-satellites, terrestrial sensors, transmitters, and micro-electromechanical systems. The power requirements are in the range of 1 mW to several hundred mW. The primary technical requirements for space applications are long life, high reliability, high specific power, and high power density, and those for some special military uses are very high power density, specific power, reliability, low radiological induced degradation, and very low radiation leakage. Thermoelectric conversion is of particular interest because of its technological maturity and proven reliability. This paper summarizes the thermoelectric, thermal, and radioisotope heat source designs and presents the corresponding performance for a number of mW size thermoelectric micro isotope power sources

  1. Microfluidic production of polymeric micro- and nanoparticles

    NARCIS (Netherlands)

    Serra, C.; Kahn, I.U.; Cortese, B.; Croon, de M.H.J.M.; Hessel, V.; Ono, T.; Anton, N.; Vandamme, Th.

    2013-01-01

    Polymeric micro- and nanoparticles have attracted a wide attention of researchers in various areas such as drug delivery, sensing, imaging, cosmetics, diagnostics, and biotechnology. However, processes with conventional equipment do not always allow a precise control of their morphology, size, size

  2. ACCESS TO MICRO CREDIT AND ECONOMIC EMPOWERMENT ...

    African Journals Online (AJOL)

    Prof

    market women have a low socio-economic status due to financial and ... market women have little or no access to micro credit schemes largely .... industry contributes to its poor performance in servicing the needs of the poor especially .... single; 11.1% are either divorced or separated; 33.3% are widows whereas a larger.

  3. Micro and nanoplatforms for biological cell analysis

    DEFF Research Database (Denmark)

    Svendsen, Winnie Edith; Castillo, Jaime; Moresco, Jacob Lange

    2010-01-01

    studies mimicking the in vivo situation is presented and an example of surface modification for cellular growth is described. Then novel electronic sensor platforms are discussed and an example of a nanosensor with electronic readout is given utilizing both micro- and nanotechnology. Finally an example...

  4. Nonlinear dynamics of biomimetic micro air vehicles

    Energy Technology Data Exchange (ETDEWEB)

    Hou, Y; Kong, J [College of Mechanical Automation, Wuhan University of Science and Technology, Wuhan, 430081 (China)], E-mail: fly_houyu@163.com.cn

    2008-02-15

    Flapping-wing micro air vehicles (FMAV) are new conceptual air vehicles that mimic the flying modes of birds and insects. They surpass the research fields of traditional airplane design and aerodynamics on application technologies, and initiate the applications of MEMS technologies on aviation fields. This paper studies a micro flapping mechanism that based upon insect thorax and actuated by electrostatic force. Because there are strong nonlinear coupling between the two physical domains, electrical and mechanical, the static and dynamic characteristics of this system are very complicated. Firstly, the nonlinear dynamic model of the electromechanical coupling system is set up according to the physical model of the flapping mechanism. The dynamic response of the system in constant voltage is studied by numerical method. Then the effect of damping and initial condition on dynamic characteristics of the system is analyzed in phase space. In addition, the dynamic responses of the system in sine voltage excitation are discussed. The results of research are helpful to the design, fabrication and application of the micro flapping mechanism of FMAV, and also to other micro electromechanical system that actuated by electrostatic force.

  5. Micro-powder injection moulding of tungsten

    International Nuclear Information System (INIS)

    Zeep, B.

    2007-12-01

    For He-cooled Divertors as integral components of future fusion power plants, about 300000 complex shaped tungsten components are to be fabricated. Tungsten is the favoured material because of its excellent properties (high melting point, high hardness, high sputtering resistance, high thermal conductivity). However, the material's properties cause major problems for large scale production of complex shaped components. Due to the resistance of tungsten to mechanical machining, new fabrication technologies have to be developed. Powder injection moulding as a well established shaping technology for a large scale production of complex or even micro structured parts might be a suitable method to produce tungsten components for fusion applications but is not yet commercially available. The present thesis is dealing with the development of a powder injection moulding process for micro structured tungsten components. To develop a suitable feedstock, the powder particle properties, the binder formulation and the solid load were optimised. To meet the requirements for a replication of micro patterned cavities, a special target was to define the smallest powder particle size applicable for micro-powder injection moulding. To investigate the injection moulding performance of the developed feedstocks, experiments were successfully carried out applying diverse cavities with structural details in micro dimension. For debinding of the green bodies, a combination of solvent debinding and thermal debinding has been adopted for injection moulded tungsten components. To develop a suitable debinding strategy, a variation of the solvent debinding time, the heating rate and the binder formulation was performed. For investigating the thermal consolidation behaviour of tungsten components, sinter experiments were carried out applying tungsten powders suitable for micro-powder injection moulding. First mechanical tests of the sintered samples showed promising material properties such as a

  6. Thermal engineering and micro-technology; Thermique et microtechnologie

    Energy Technology Data Exchange (ETDEWEB)

    Kandlikar, S. [Rochester Inst. of Tech., NY (United States); Luo, L. [Institut National Polytechnique, 54 - Nancy (France); Gruss, A. [CEA Grenoble, GRETH, 38 (France); Wautelet, M. [Mons Univ. (Belgium); Gidon, S. [CEA Grenoble, Lab. d' Electronique et de Technologie de l' Informatique (LETI), 38 (France); Gillot, C. [Ecole Nationale Superieure d' Ingenieurs Electriciens de Grenoble, 38 - Saint Martin d' Heres (France)]|[CEA Grenoble, Lab. Electronique et de Technologie de l' Informatique (LETI), 38 (France); Therme, J.; Marvillet, Ch.; Vidil, R. [CEA Grenoble, 38 (France); Dutartre, D. [ST Microelectronique, France (France); Lefebvre, Ph. [SNECMA, 75 - Paris (France); Lallemand, M. [Institut National des Sciences Appliquees (INSA), 69 - Villeurbanne (France); Colin, S. [Institut National des Sciences Appliquees (INSA), 31 - Toulouse (France); Joulin, K. [Ecole Nationale Superieure de Mecanique et d' Aerotechnique (ENSMA), 86 - Poitiers (France); Gad el Hak, M. [Virginia Univ., Charlottesville, VA (United States)

    2003-07-01

    This document gathers the abstracts and transparencies of 5 invited conferences of this congress of the SFT about heat transfers and micro-technologies: Flow boiling in microchannels: non-dimensional groups and heat transfer mechanisms (S. Kandlikar); Intensification and multi-scale process units (L. Luo and A. Gruss); Macro-, micro- and nano-systems: different physics? (M. Wautelet); micro-heat pipes (M. Lallemand); liquid and gas flows inside micro-ducts (S. Colin). The abstracts of the following presentations are also included: Electro-thermal writing of nano-scale memory points in a phase change material (S. Gidon); micro-technologies for cooling in micro-electronics (C. Gillot); the Minatec project (J. Therme); importance and trends of thermal engineering in micro-electronics (D. Dutartre); Radiant heat transfers at short length scales (K. Joulain); Momentum and heat transfer in micro-electromechanical systems (M. Gad-el-Hak). (J.S.)

  7. Nonequilibrium fluctuations in micro-MHD effects on electrodeposition

    International Nuclear Information System (INIS)

    Aogaki, Ryoichi; Morimoto, Ryoichi; Asanuma, Miki

    2010-01-01

    In copper electrodeposition under a magnetic field parallel to electrode surface, different roles of two kinds of nonequilibrium fluctuations for micro-magnetohydrodynamic (MHD) effects are discussed; symmetrical fluctuations are accompanied by the suppression of three dimensional (3D) nucleation by micro-MHD flows (the 1st micro-MHD effect), whereas asymmetrical fluctuations controlling 2D nucleation yield secondary nodules by larger micro-MHD flows (the 2nd micro-MHD effect). Though the 3D nucleation with symmetrical fluctuations is always suppressed by the micro-MHD flows, due to the change in the rate-determining step from electron transfer to mass transfer, the 2D nucleation with asymmetrical fluctuations newly turns unstable, generating larger micro-MHD flows. As a result, round semi-spherical deposits, i.e., secondary nodules are yielded. Using computer simulation, the mechanism of the 2nd micro-MHD effect is validated.

  8. influence of some variable parameters on horizontal elliptic micro

    African Journals Online (AJOL)

    temidayo

    2013-07-02

    Jul 2, 2013 ... 1,3,4DEPARTMENT OF MECHANICAL ENGINEERING, FACULTY OF ENGINEERING AND ... Keywords: Ellipse, Micro-channels, internal fins, Heat transfer, Fluid flow ...... reaction in micro scale segmented gas-liquid.

  9. Micro-Combined Heat and Power Device Test Facility

    Data.gov (United States)

    Federal Laboratory Consortium — NIST has developed a test facility for micro-combined heat and power (micro-CHP) devices to measure their performance over a range of different operating strategies...

  10. MicroRNAs as regulatory elements in psoriasis

    Directory of Open Access Journals (Sweden)

    Liu Yuan

    2016-01-01

    Full Text Available Psoriasis is a chronic, autoimmune, and complex genetic disorder that affects 23% of the European population. The symptoms of Psoriatic skin are inflammation, raised and scaly lesions. microRNA, which is short, nonprotein-coding, regulatory RNAs, plays critical roles in psoriasis. microRNA participates in nearly all biological processes, such as cell differentiation, development and metabolism. Recent researches reveal that multitudinous novel microRNAs have been identified in skin. Some of these substantial novel microRNAs play as a class of posttranscriptional gene regulator in skin disease, such as psoriasis. In order to insight into microRNAs biological functions and verify microRNAs biomarker, we review diverse references about characterization, profiling and subtype of microRNAs. Here we will share our opinions about how and which microRNAs are as regulatory in psoriasis.

  11. ICTs and Urban Micro Enterprises : Maximizing Opportunities for ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    ICTs and Urban Micro Enterprises : Maximizing Opportunities for Economic Development ... the use of ICTs in micro enterprises and their role in reducing poverty. ... in its approach to technological connectivity but bottom-up in relation to.

  12. The micro turbine: the MIT example; La micro turbine: l'exemple du MIT

    Energy Technology Data Exchange (ETDEWEB)

    Ribaud, Y. [Office National d' Etudes et de Recherches Aerospatiales (ONERA-DEFA), 92 - Chatillon (France)

    2001-10-01

    The micro turbine study began a few years ago at the MIT, with the participation of specialists from different fields. The purpose is the development of a MEMS (micro electro mechanical systems) based, 1 cm in diameter, micro gas turbine. Potential applications are devoted to micro drone propulsion, electric power generation for portable power sources in order to replace heavy Lithium batteries, satellite motorization, the surface distributed power for boundary suction on plane wings. The manufacturing constraints at such small scales lead to 2-D extruded shapes. The physical constraints stem from viscous effects and from limitations given by 2-D geometry. The time scales are generally shorter than for conventional machines. Otherwise the material properties are better at such length scales. Transposition from conventional turbomachinery laws is no more applicable and new design methods must be established. The present paper highlights the project progress and the technology breakthroughs. (author)

  13. Doppler speedometer for micro-organisms

    International Nuclear Information System (INIS)

    Penkov, F.; Tuleushev, A.; Lisitsyn, V.; Kim, S.; Tuleushev, Yu.

    1996-01-01

    Objective of Investigations: Development and creation of the Doppler speedometer for micro-organisms which allows to evaluate, in a real temporal scale, variations in the state of water suspension of micro-organisms under the effect of chemical, physical and other external actions. Statement of the Problem The main problem is absence of reliable, accessible for users and simple, in view of application, Doppler speedometers for micro-organisms. Nevertheless, correlation Doppler spectrometry in the regime of heterodyning the supporting and cell-scattered laser radiation is welt known. The main idea is that the correlation function of photo-current pulses bears an information on the averages over the assembly of cell velocities. For solving the biological problems, construction of auto-correlation function in the real-time regime with the delay time values comprising, function in the real-time regime with the delay time values comprising, nearly, 100 me (10 khz) or higher is needed. Computers of high class manage this problem using but the program software. Due to this, one can simplify applications of the proposed techniques provided he creates the Doppler speedometer for micro-organism on a base of the P entium . Expected Result Manufactured operable mock-up of the Doppler speedometer for micro-organisms in a form of the auxiliary computer block which allows to receive an information, in the real time scale, on the results of external effects of various nature on the cell assembly in transparent medium with a small volume of the studied cell suspension

  14. Primary expatiation on micro material evidence and authentication

    International Nuclear Information System (INIS)

    Yang Mingtai; Wang Wen; Wu Lunqiang; Dai Changsong

    2012-01-01

    The micro material evidence is the impersonal and concrete material evidences, and the quantity and volume is small, but it plays an important role in judicature litigation. In the paper, the basic character, type and form mechanism of the micro material evidence have been analyzed and discussed, and the gist of the micro material evidence has been summarized. It must play a helpful role in the micro material evidence to correlative workers for cognizance and application. (authors)

  15. Brushless DC micro-motor with external rotor

    International Nuclear Information System (INIS)

    Rizzo, M.; Turowski, J.

    1992-01-01

    The increasing use of high-tech electronic components has led researchers to try new solutions in the field of micro-scale electrical machinery. One such solution, described in this paper, consists of the substitution of a conventional mechanical commutator with an electronic type so as to allow the conversion of a electromagnetic micro-motor into a brushless version using permanent magnets. The comparison of the two micro-motor alternatives evidences the clear superiority of the brushless micro-motor

  16. Thermal conductivity of microPCMs-filled epoxy matrix composites

    OpenAIRE

    Su, J.F.; Wang, X.Y; Huang, Z.; Zhao, Y.H.; Yuan, X.Y.

    2011-01-01

    Microencapsulated phase change materials (microPCMs) have been widely applied in solid matrix as thermal-storage or temperature-controlling functional composites. The thermal conductivity of these microPCMs/matrix composites is an important property need to be considered. In this study, a series of microPCMs have been fabricated using the in situ polymerization with various core/shell ratio and average diameter; the thermal conductivity of microPCMs/epoxy composites were investigated in detai...

  17. Replacement power supply with micro-hydropower. Case micro central - Pipinta

    International Nuclear Information System (INIS)

    Gomez, Jorge I; Hincapie, Luis A; Woodcock, Edgar; Arregoces Alvaro

    2006-01-01

    This paper describes the Micro-Hydro Electric Pipinta through its main components and their parameters. An evaluation of the actual power plant costs and a proposed redesign are also given with prices referenced to year 2004. A comparison between the electricity price supplied by the grid and the cost for the electricity in this Micro-Hydro Power Plant let conclude that projects of this type are available in rural areas of Colombia reached by the grid.

  18. The Model of Optimization of Micro Energy; HOMER: El Modelo de Optimizacin de Micro energa

    Energy Technology Data Exchange (ETDEWEB)

    2004-05-01

    HOMER, the model of optimization of micro energy, helps to disear systems out of the network and interconnected to the network. You can use HOMER to carry out the analysis to explore an extensive rank of questions of diseo. HOMER, el modelo de optimizacin de micro energa, le ayuda a disear sistemas fuera de la red e interconectados a la red. Usted puede usar HOMER para llevar a cabo el anlisis para explorar un amplio rango de preguntas de diseo.

  19. Determination of elemental abundances in impact materials by micro-PIXE and micro-SRXRF methods

    International Nuclear Information System (INIS)

    Uzonyi, I.; Szabo, Gy.; Kiss, A.Z.; Szoeoer, Gy.; Rozsa, P.

    2004-01-01

    The most famous and well-preserved meteorite crater in the world is the Barringer Meteor Crater (Arizona, USA). The meteorite is supposed to be a fragment of a small asteroid of our solar system. During the impact event the matter of the projectile mixed with that of the target rocks forming breccias, slag and spherules. For the non-destructive characterization of the impact materials a combined micro-PIXE and micro-SRXRF technique was applied. (N.T.)

  20. DIANA-microT web server: elucidating microRNA functions through target prediction.

    Science.gov (United States)

    Maragkakis, M; Reczko, M; Simossis, V A; Alexiou, P; Papadopoulos, G L; Dalamagas, T; Giannopoulos, G; Goumas, G; Koukis, E; Kourtis, K; Vergoulis, T; Koziris, N; Sellis, T; Tsanakas, P; Hatzigeorgiou, A G

    2009-07-01

    Computational microRNA (miRNA) target prediction is one of the key means for deciphering the role of miRNAs in development and disease. Here, we present the DIANA-microT web server as the user interface to the DIANA-microT 3.0 miRNA target prediction algorithm. The web server provides extensive information for predicted miRNA:target gene interactions with a user-friendly interface, providing extensive connectivity to online biological resources. Target gene and miRNA functions may be elucidated through automated bibliographic searches and functional information is accessible through Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The web server offers links to nomenclature, sequence and protein databases, and users are facilitated by being able to search for targeted genes using different nomenclatures or functional features, such as the genes possible involvement in biological pathways. The target prediction algorithm supports parameters calculated individually for each miRNA:target gene interaction and provides a signal-to-noise ratio and a precision score that helps in the evaluation of the significance of the predicted results. Using a set of miRNA targets recently identified through the pSILAC method, the performance of several computational target prediction programs was assessed. DIANA-microT 3.0 achieved there with 66% the highest ratio of correctly predicted targets over all predicted targets. The DIANA-microT web server is freely available at www.microrna.gr/microT.

  1. MicroRNA-target binding structures mimic microRNA duplex structures in humans.

    Directory of Open Access Journals (Sweden)

    Xi Chen

    Full Text Available Traditionally, researchers match a microRNA guide strand to mRNA sequences using sequence comparisons to predict its potential target genes. However, many of the predictions can be false positives due to limitations in sequence comparison alone. In this work, we consider the association of two related RNA structures that share a common guide strand: the microRNA duplex and the microRNA-target binding structure. We have analyzed thousands of such structure pairs and found many of them share high structural similarity. Therefore, we conclude that when predicting microRNA target genes, considering just the microRNA guide strand matches to gene sequences may not be sufficient--the microRNA duplex structure formed by the guide strand and its companion passenger strand must also be considered. We have developed software to translate RNA binding structure into encoded representations, and we have also created novel automatic comparison methods utilizing such encoded representations to determine RNA structure similarity. Our software and methods can be utilized in the other RNA secondary structure comparisons as well.

  2. Radar micro-doppler signatures processing and applications

    CERN Document Server

    Chen, Victor C; Miceli, William J

    2014-01-01

    Radar Micro-Doppler Signatures: Processing and applications concentrates on the processing and application of radar micro-Doppler signatures in real world situations, providing readers with a good working knowledge on a variety of applications of radar micro-Doppler signatures.

  3. Methods for the measurement of micro material evidence

    International Nuclear Information System (INIS)

    Yang Mingtai; Wang Wen; Wu Lunqiang; Dai Changsong

    2012-01-01

    The micro material evidence has been used in judicature litigation successfully, that must base on impersonal authentication with the advanced and proper methods. The basic principle, main trait, applicable range, analysis minimum and sensitive limit of twelve type instruments used in micro material evidence analysis have been related. Those could supply optional proper methods for micro material evidence. (authors)

  4. Micro digital sun sensor: system in a package

    NARCIS (Netherlands)

    Boom, C.W. de; Leijtens, J.A.P.; Duivenbode, L.M.H. van; Heiden, N. van der

    2004-01-01

    A novel micro Digital Sun Sensor (μDSS) is under development in the frame of a micro systems technology (MST) development program (Microned) from the Dutch Ministry of Economic Affairs. Use of available micro system technologies in combination with the implementation of a dedicated solarcell for

  5. Informal Micro-Enterprises and Solid Waste Collection: The Case ...

    African Journals Online (AJOL)

    The research used both Primary and secondary data sources. One hundred sixty micro-enterprise units were included in the survey. These account for about 35% of the total micro-enterprises available in the city. Stratified random sampling was employed based on the number and type of micro-enterprises available in each ...

  6. MicroRNAs: role and therapeutic targets in viral hepatitis

    NARCIS (Netherlands)

    van der Ree, Meike H.; de Bruijne, Joep; Kootstra, Neeltje A.; Jansen, Peter Lm; Reesink, Hendrik W.

    2014-01-01

    MicroRNAs regulate gene expression by binding to the 3'-untranslated region (UTR) of target messenger RNAs (mRNAs). The importance of microRNAs has been shown for several liver diseases, for example, viral hepatitis. MicroRNA-122 is highly abundant in the liver and is involved in the regulation of

  7. Radar Micro-Doppler classification of Mini-UAVs

    NARCIS (Netherlands)

    Harmanny, R.L.A.; Prémel-Cabic, G.; Wit, J.J.M.

    2015-01-01

    The radar micro-Doppler signature of a target depends on its micro-motion, i.e., the motion of parts of a target relative to the motion of the target as a whole. These micro-motions are very characteristic considering different target classes, e.g., the slow pendulum-like motion of a bird’s wings

  8. Micro-processus et macro-structures

    Directory of Open Access Journals (Sweden)

    Aaron Victor Cicourel

    2008-10-01

    Full Text Available Des approches sociologiques traditionnelles ont défini des macro-structures sociales comme un niveau particulier de la réalité sociale, à distinguer des micro-épisodes de l’action sociale. Cela les a conduits à concevoir ces macro-structures et à mener des recherches sur elles de manière plus ou moins indépendante des pratiques observables de la vie quotidienne. Cicourel soutient que les faits (macro-sociaux ne sont pas simplement donnés, mais émergent de pratiques routinières de la vie de tous les jours. Le macro, au sens de descriptions résumées, hors contexte, normalisées et typifiées, est un produit typique des procédures interactives et organisationnelles qui transforment les micro-événements en structures macro-sociales. Ainsi une précondition pour l’intégration des phénomènes micro- et macro-sociaux dans notre théorie et dans notre méthodologie renvoie à l’identification des processus contribuant à la création de macro-structures par des inférences routinières, des interprétations et des procédure de résumé. Le texte montre aussi que les différences entre approches micro-sociologiques apparaissent parallèles à celles existant entre approches micro et macro. On se centrant sur de petits fragments d’interactions conversationnelles, certains travaux micro-sociologiques tendent à ignorer ce qui informe ces interactions conversationnelles pour les participants eux-mêmes. Les comptes rendus décontextualisés produits par de telles méthodes ressemblent à la décontextualisation résultant des procédures macro-sociologiques d’agrégation. Contre cela, Cicourel défend la constitution de bases de données comparatives n’incluant pas seulement le contexte des interactions de face à face, mais étudiant aussi les phénomènes sociaux de manière systématique à travers différents contextes.Micro-processes and macro-structures. Notes on articulation between different levels of analysis

  9. Micro-CHP Systems for Residential Applications

    Energy Technology Data Exchange (ETDEWEB)

    Timothy DeValve; Benoit Olsommer

    2007-09-30

    Integrated micro-CHP (Cooling, Heating and Power) system solutions represent an opportunity to address all of the following requirements at once: conservation of scarce energy resources, moderation of pollutant release into our environment, and assured comfort for home-owners. The objective of this effort was to establish strategies for development, demonstration, and sustainable commercialization of cost-effective integrated CHP systems for residential applications. A unified approach to market and opportunity identification, technology assessment, specific system designs, adaptation to modular product platform component conceptual designs was employed. UTRC's recommendation to U.S. Department of Energy is to go ahead with the execution of the proposed product development and commercialization strategy plan under Phase II of this effort. Recent indicators show the emergence of micro-CHP. More than 12,000 micro-CHP systems have been sold worldwide so far, around 7,500 in 2004. Market projections predict a world-wide market growth over 35% per year. In 2004 the installations were mainly in Europe (73.5%) and in Japan (26.4%). The market in North-America is almost non-existent (0.1%). High energy consumption, high energy expenditure, large spark-spread (i.e., difference between electricity and fuel costs), big square footage, and high income are the key conditions for market acceptance. Today, these conditions are best found in the states of New York, Pennsylvania, New Jersey, Wisconsin, Illinois, Indiana, Michigan, Ohio, New England states. A multiple stage development plan is proposed to address risk mitigation. These stages include concept development and supplier engagement, component development, system integration, system demonstration, and field trials. A two stage commercialization strategy is suggested based on two product versions. The first version--a heat and power system named Micro-Cogen, provides the heat and essential electrical power to the

  10. Power and hydrogen production from ammonia in a micro-thermophotovoltaic device integrated with a micro-reformer

    International Nuclear Information System (INIS)

    Um, Dong Hyun; Kim, Tae Young; Kwon, Oh Chae

    2014-01-01

    Power and hydrogen (H 2 ) production by burning and reforming ammonia (NH 3 ) in a micro-TPV (microscale-thermophotovoltaic) device integrated with a micro-reformer is studied experimentally. A heat-recirculating micro-emitter with the cyclone and helical adapters that enhance the residence time of fed fuel-air mixtures and uniform burning burns H 2 -added NH 3 -air mixtures. A micro-reformer that converts NH 3 to H 2 using ruthenium as a catalyst surrounds the micro-emitter as a heat source. The micro-reformer is surrounded by a chamber, the inner and outer walls of which have installations of gallium antimonide photovoltaic cells and cooling fins. For the micro-reformer-integrated micro-TPV device the maximum overall efficiency of 8.1% with electrical power of 4.5 W and the maximum NH 3 conversion rate of 96.0% with the H 2 production rate of 22.6 W (based on lower heating value) are obtained, indicating that the overall efficiency is remarkably enhanced compared with 2.0% when the micro-TPV device operates alone. This supports the potential of improving the overall efficiency of a micro-TPV device through integrating it with a micro-reformer. Also, the feasibility of using NH 3 as a carbon-free fuel for both burning and reforming in practical micro power and H 2 generation devices has been demonstrated. - Highlights: • Performance of micro-TPV device integrated with micro-reformer is evaluated. • Feasibility of using NH 3 –H 2 blends in integrated system has been demonstrated. • Integration with micro-reformer improves performance of micro-TPV device. • Maximum overall efficiency of 8.1% is found compared with 2.0% without integration

  11. Study for increasing micro-drill reliability by vibrating drilling

    International Nuclear Information System (INIS)

    Yang Zhaojun; Li Wei; Chen Yanhong; Wang Lijiang

    1998-01-01

    A study for increasing micro-drill reliability by vibrating drilling is described. Under the experimental conditions of this study it is observed, from reliability testing and the fitting of a life-distribution function, that the lives of micro-drills under ordinary drilling follow the log-normal distribution and the lives of micro-drills under vibrating drilling follow the Weibull distribution. Calculations for reliability analysis show that vibrating drilling can increase the lives of micro-drills and correspondingly reduce the scatter of drill lives. Therefore, vibrating drilling increases the reliability of micro-drills

  12. Gas Mixtures for Welding with Micro-Jet Cooling

    OpenAIRE

    Węgrzyn T.

    2015-01-01

    Welding with micro-jet cooling after was tested only for MIG and MAG processes. For micro-jet gases was tested only argon, helium and nitrogen. A paper presents a piece of information about gas mixtures for micro-jet cooling after in welding. There are put down information about gas mixtures that could be chosen both for MAG welding and for micro-jet process. There were given main information about influence of various micro-jet gas mixtures on metallographic structure of steel welds. Mechani...

  13. MicroRNA expression profiling of the porcine developing brain

    DEFF Research Database (Denmark)

    Podolska, Agnieszka; Kaczkowski, Bogumil; Busk, Peter Kamp

    2011-01-01

    MicroRNAs are small, non-coding RNA molecules that regulate gene expression at the post-transcriptional level and play an important role in the control of developmental and physiological processes. In particular, the developing brain contains an impressive diversity of microRNAs. Most micro...... and the growth curve when compared to humans. Considering these similarities, studies examining microRNA expression during porcine brain development could potentially be used to predict the expression profile and role of microRNAs in the human brain....

  14. Sequential micro and ultrafiltration of distillery wastewater

    Directory of Open Access Journals (Sweden)

    Vasić Vesna M.

    2015-01-01

    Full Text Available Water reuse and recycling, wastewater treatment, drinking water production and environmental protection are the key challenges for the future of our planet. Membrane separation technologies for the removal of all suspended solids and a fraction of dissolved solids from wastewaters, are becoming more and more promising. Also, these processes are playing a major role in wastewater purification systems because of their high potential for recovery of water from many industrial wastewaters. The aim of this work was to evaluate the application of micro and ultrafiltration for distillery wastewater purification in order to produce water suitable for reuse in the bioethanol industry. The results of the analyses of the permeate obtained after micro and ultrafiltration showed that the content of pollutants in distillery wastewater was significantly reduced. The removal efficiency for chemical oxygen demand, dry matter and total nitrogen was 90%, 99.2% and 99.9%, respectively. Suspended solids were completely removed from the stillage.

  15. Verification of Tolerance Chains in Micro Manufacturing

    DEFF Research Database (Denmark)

    Gasparin, Stefania

    .1 – 200 μm). Finally, an optical component is investigated with the purpose of suggesting a quality control approach for micro-manufacturing process through a control of the product. It is a useful method to adopt when the aim is to detect and quantify inconsistency or incompatibilities during a process...... on dimensional and geometrical metrology. If the measurements uncertainty is large compared to the tolerance interval, a small conformance zone is left for process variation. Therefore particular attention has to be paid to the instrument capabilities in order to reduce the measurement uncertainty. Different...... chain. In this way the process parameters can be adjusted in order to fulfil the requirements of the final micro-product....

  16. Vision-guidance of micro- and nanorobots

    Science.gov (United States)

    Wortmann, T.; Dahmen, C.; Fatikow, S.

    2012-01-01

    This paper presents a selection of image processing methods and algorithms, which are needed to enable the reliable automation of robotic tasks at the micro and nanoscale. Application examples are automatic assembly of new nanoscale electronic elements or automatic testing of material properties. Due to the very small object dimensions targeted here, the scanning electron microscope is the appropriate image sensor. The methods described in this paper can be categorized into procedures of object recognition and object tracking. Object recognition deals with the problem of finding and labeling nanoscale objects in an image scene, whereas tracking is the process of continuously following the movement of a specific object. Both methods carried out subsequently enable fully automated robotic tasks at the micro- and nanoscale. A selection of algorithms is demonstrated and found suitable.

  17. Micro-system inertial sensing technology overview.

    Energy Technology Data Exchange (ETDEWEB)

    Allen, James Joe

    2009-02-01

    The purpose of this report is to provide an overview of Micro-System technology as it applies to inertial sensing. Transduction methods are reviewed with capacitance and piezoresistive being the most often used in COTS Micro-electro-mechanical system (MEMS) inertial sensors. Optical transduction is the most recent transduction method having significant impact on improving sensor resolution. A few other methods are motioned which are in a R&D status to hopefully allow MEMS inertial sensors to become viable as a navigation grade sensor. The accelerometer, gyroscope and gravity gradiometer are the type of inertial sensors which are reviewed in this report. Their method of operation and a sampling of COTS sensors and grade are reviewed as well.

  18. Energy Conversion at Micro and Nanoscale

    International Nuclear Information System (INIS)

    Gammaitoni, Luca

    2014-01-01

    Energy management is considered a task of strategic importance in contemporary society. It is a common fact that the most successful economies of the planet are the economies that can transform and use large quantities of energy. In this talk we will discuss the role of energy with specific attention to the processes that happens at micro and nanoscale. The description of energy conversion processes at these scales requires approaches that go way beyond the standard equilibrium termodynamics of macroscopic systems. In this talk we will address from a fundamental point of view the physics of the dissipation of energy and will focus our attention to the energy transformation processes that take place in the modern micro and nano information and communication devices

  19. MIDN: A spacecraft Micro-dosimeter mission

    International Nuclear Information System (INIS)

    Pisacane, V. L.; Ziegler, J. F.; Nelson, M. E.; Caylor, M.; Flake, D.; Heyen, L.; Youngborg, E.; Rosenfeld, A. B.; Cucinotta, F.; Zaider, M.; Dicello, J. F.

    2006-01-01

    MIDN (Micro-dosimetry instrument) is a payload on the MidSTAR-I spacecraft (Midshipman Space Technology Applications Research) under development at the United States Naval Academy. MIDN is a solid-state system being designed and constructed to measure Micro-dosimetric spectra to determine radiation quality factors for space environments. Radiation is a critical threat to the health of astronauts and to the success of missions in low-Earth orbit and space exploration. The system will consist of three separate sensors, one external to the spacecraft, one internal and one embedded in polyethylene. Design goals are mass <3 kg and power <2 W. The MidSTAR-I mission in 2006 will provide an opportunity to evaluate a preliminary version of this system. Its low power and mass makes it useful for the International Space Station and manned and unmanned interplanetary missions as a real-time system to assess and alert astronauts to enhanced radiation environments. (authors)

  20. Micro Z - in nuclear power generation

    International Nuclear Information System (INIS)

    Anon.

    1984-01-01

    Micro-Z microprocessor boiler control systems have been adopted by the French State Electricity utility for their next group of 1300MW units. It replaces the analogue system used in earlier units. Micro-Z is a fully distributed digital control and regulation system and in contrast to earlier digital systems with centralised data processing, processing is carried out in localised sub-units such as regulators and actuators. In the system adopted by EdF for the 1300MW units, digital communication with centralised and common control stations and supervisory computer were not required. The system is made up of control cards, interface modules, separate stations and configuration and control hardware and software. There are no fundamental difference in general system design between analogue and digital systems, the differences are found in digitization of signals and concentration of processing at board level. (U.K.)

  1. Lignin from Micro- to Nanosize: Applications

    Directory of Open Access Journals (Sweden)

    Stefan Beisl

    2017-11-01

    Full Text Available Micro- and nanosize lignin has recently gained interest due to improved properties compared to standard lignin available today. As the second most abundant biopolymer after cellulose, lignin is readily available but used for rather low-value applications. This review focuses on the application of micro- and nanostructured lignin in final products or processes that all show potential for high added value. The fields of application are ranging from improvement of mechanical properties of polymer nanocomposites, bactericidal and antioxidant properties and impregnations to hollow lignin drug carriers for hydrophobic and hydrophilic substances. Also, a carbonization of lignin nanostructures can lead to high-value applications such as use in supercapacitors for energy storage. The properties of the final product depend on the surface properties of the nanomaterial and, therefore, on factors like the lignin source, extraction method, and production/precipitation methods, as discussed in this review.

  2. Mini and Micro Propulsion for Medical Swimmers

    Directory of Open Access Journals (Sweden)

    JianFeng

    2014-02-01

    Full Text Available Mini and micro robots, which can swim in an underwater environment, have drawn widespread research interests because of their potential applicability to the medical or biological fields, including delivery and transportation of bio-materials and drugs, bio-sensing, and bio-surgery. This paper reviews the recent ideas and developments of these types of self-propelling devices, ranging from the millimeter scale down to the micro and even the nano scale. Specifically, this review article makes an emphasis on various propulsion principles, including methods of utilizing smart actuators, external magnetic/electric/acoustic fields, bacteria, chemical reactions, etc. In addition, we compare the propelling speed range, directional control schemes, and advantages of the above principles.

  3. Biological stoichiometry in tumor micro-environments.

    Directory of Open Access Journals (Sweden)

    Irina Kareva

    Full Text Available Tumors can be viewed as evolving ecological systems, in which heterogeneous populations of cancer cells compete with each other and somatic cells for space and nutrients within the ecosystem of the human body. According to the growth rate hypothesis (GRH, increased phosphorus availability in an ecosystem, such as the tumor micro-environment, may promote selection within the tumor for a more proliferative and thus potentially more malignant phenotype. The applicability of the GRH to tumor growth is evaluated using a mathematical model, which suggests that limiting phosphorus availability might promote intercellular competition within a tumor, and thereby delay disease progression. It is also shown that a tumor can respond differently to changes in its micro-environment depending on the initial distribution of clones within the tumor, regardless of its initial size. This suggests that composition of the tumor as a whole needs to be evaluated in order to maximize the efficacy of therapy.

  4. MICRO-CHP System for Residential Applications

    Energy Technology Data Exchange (ETDEWEB)

    Joseph Gerstmann

    2009-01-31

    This is the final report of progress under Phase I of a project to develop and commercialize a micro-CHP system for residential applications that provides electrical power, heating, and cooling for the home. This is the first phase of a three-phase effort in which the residential micro-CHP system will be designed (Phase I), developed and tested in the laboratory (Phase II); and further developed and field tested (Phase III). The project team consists of Advanced Mechanical Technology, Inc. (AMTI), responsible for system design and integration; Marathon Engine Systems, Inc. (MES), responsible for design of the engine-generator subsystem; AO Smith, responsible for design of the thermal storage and water heating subsystems; Trane, a business of American Standard Companies, responsible for design of the HVAC subsystem; and AirXchange, Inc., responsible for design of the mechanical ventilation and dehumidification subsystem.

  5. Micro-separation toward systems biology.

    Science.gov (United States)

    Liu, Bi-Feng; Xu, Bo; Zhang, Guisen; Du, Wei; Luo, Qingming

    2006-02-17

    Current biology is experiencing transformation in logic or philosophy that forces us to reevaluate the concept of cell, tissue or entire organism as a collection of individual components. Systems biology that aims at understanding biological system at the systems level is an emerging research area, which involves interdisciplinary collaborations of life sciences, computational and mathematical sciences, systems engineering, and analytical technology, etc. For analytical chemistry, developing innovative methods to meet the requirement of systems biology represents new challenges as also opportunities and responsibility. In this review, systems biology-oriented micro-separation technologies are introduced for comprehensive profiling of genome, proteome and metabolome, characterization of biomolecules interaction and single cell analysis such as capillary electrophoresis, ultra-thin layer gel electrophoresis, micro-column liquid chromatography, and their multidimensional combinations, parallel integrations, microfabricated formats, and nano technology involvement. Future challenges and directions are also suggested.

  6. Cavity Pressure Behaviour in Micro Injection Moulding

    DEFF Research Database (Denmark)

    Griffiths, C.A.; Dimov, S.S.; Scholz, S.

    2010-01-01

    as well as with the filling of the cavity by the polymer melt. In this paper, two parameters derived from cavity pressure over time (i.e. pressure work). The influence of four µIM parameters (melt temperature, mould temperature, injection speed, aand packing pressure) on the two pressure-related outputs...... has been investigated by moulding a micro fluidic component on three different polymers (PP, ABS, PC) using the design of experiment approach. Similar trends such as the effects of a higher injection speed in decreasing the pressure work and of a lower temperature in decreasing pressure rate have been......Process monitoring of micro injection moulding (µIM) is of crusial importance to analyse the effect of different parameter settings on the process and to assess its quality. Quality factors related to cavity pressure can provide useful information directly connected with the dyanmics of the process...

  7. Micro-generation network connection (renewables)

    Energy Technology Data Exchange (ETDEWEB)

    Thornycroft, J.; Russell, T.; Curran, J.

    2003-07-01

    The drive to reduce emissions of carbon dioxide will result in an increase in the number of small generation units seeking connection to the electric power distribution network. The objectives of this study were to consider connection issues relating to micro-generation from renewables and their integration into the UK distribution network. The document is divided into two sections. The first section describes the present system which includes input from micro-generation, the technical impacts and the financial considerations. The second part discusses technical, financial and governance options for the future. A summary of preferred options and recommendations is given. The study was carried out by the Halcrow Group Ltd under contract to the DTI.

  8. Forming of Polymeric Tubular Micro-components

    DEFF Research Database (Denmark)

    Qin, Yi; Zhao, Jie; Anyasodor, Gerald

    2015-01-01

    platform for the production of functional polymeric tubular micro-components. The chapter gives background on the current market and process development trends, followed by description of materials, process configuration, tool design and machine development for each processing technology as well......This chapter is intended to provide an overview of three nontraditional shaping technologies for the forming of polymeric micro-tubes, which are hot embossing, blow molding, and cross rolling, as well as realization of a process chain and the integration of a modular machine-based manufacturing...... as strategy for integration of the technologies and equipment into a common platform. Finally, potential applications of the technologies and facilities developed are highlighted....

  9. Deformations in micro extrusion of metals

    Directory of Open Access Journals (Sweden)

    J. Piwnik

    2010-07-01

    Full Text Available Production technologies of small dimensions metallic elements are known for a long time. They are produced by machining methods:turning, milling, polishing. Recently, methods for manufacturing small details by forming are developed – microforming. This process ischaracterized by the high dimensions accuracy and the surface smoothness of received items and the high production rate. When a forming process is scaled down to micro dimensions, the microstructure of the workpiece, the surface topology of the workpiece and that of the tooling remain unchanged. Size effect is appearing. This paper analyses specifications of a metal extrusion in micro scale. To determine the impact of the tool surface roughness on deformation process the numerical model of roughness as triangle wave were developed. In paper the influence of the wave presence on the material flow is described. Impact of the forming conditions on extrusion forces there is also characterized.

  10. Payment mechanisms for micro-generation

    Energy Technology Data Exchange (ETDEWEB)

    Choudhury, W.; Andrews, S.

    2002-07-01

    The Department of Trade and Industry commissioned a study into payment options for the increasing number of micro-generators (in domestic dwellings and where the generating capacity is not more than 5kW i.e. essentially micro-CHP and photovoltaics) supplying power for the national distribution network. It is shown that small generators will impact on all aspects of industry and connection will need to be simplified. The network will call for a more actively managed regime. Metering will need to be more sophisticated and agreement reached on how costs should be allocated when demand is low and surplus electricity is exported to the system. Three options were considered to be viable.

  11. Machine learning for micro-tomography

    Science.gov (United States)

    Parkinson, Dilworth Y.; Pelt, Daniël. M.; Perciano, Talita; Ushizima, Daniela; Krishnan, Harinarayan; Barnard, Harold S.; MacDowell, Alastair A.; Sethian, James

    2017-09-01

    Machine learning has revolutionized a number of fields, but many micro-tomography users have never used it for their work. The micro-tomography beamline at the Advanced Light Source (ALS), in collaboration with the Center for Applied Mathematics for Energy Research Applications (CAMERA) at Lawrence Berkeley National Laboratory, has now deployed a series of tools to automate data processing for ALS users using machine learning. This includes new reconstruction algorithms, feature extraction tools, and image classification and recommen- dation systems for scientific image. Some of these tools are either in automated pipelines that operate on data as it is collected or as stand-alone software. Others are deployed on computing resources at Berkeley Lab-from workstations to supercomputers-and made accessible to users through either scripting or easy-to-use graphical interfaces. This paper presents a progress report on this work.

  12. Micro-inverter solar panel mounting

    Science.gov (United States)

    Morris, John; Gilchrist, Phillip Charles

    2016-02-02

    Processes, systems, devices, and articles of manufacture are provided. Each may include adapting micro-inverters initially configured for frame-mounting to mounting on a frameless solar panel. This securement may include using an adaptive clamp or several adaptive clamps secured to a micro-inverter or its components, and using compressive forces applied directly to the solar panel to secure the adaptive clamp and the components to the solar panel. The clamps can also include compressive spacers and safeties for managing the compressive forces exerted on the solar panels. Friction zones may also be used for managing slipping between the clamp and the solar panel during or after installation. Adjustments to the clamps may be carried out through various means and by changing the physical size of the clamps themselves.

  13. High-speed micro-electro-discharge machining.

    Energy Technology Data Exchange (ETDEWEB)

    Chandrasekar, Srinivasan Dr. (.School of Industrial Engineering, West Lafayette, IN); Moylan, Shawn P. (School of Industrial Engineering, West Lafayette, IN); Benavides, Gilbert Lawrence

    2005-09-01

    When two electrodes are in close proximity in a dielectric liquid, application of a voltage pulse can produce a spark discharge between them, resulting in a small amount of material removal from both electrodes. Pulsed application of the voltage at discharge energies in the range of micro-Joules results in the continuous material removal process known as micro-electro-discharge machining (micro-EDM). Spark erosion by micro-EDM provides significant opportunities for producing small features and micro-components such as nozzle holes, slots, shafts and gears in virtually any conductive material. If the speed and precision of micro-EDM processes can be significantly enhanced, then they have the potential to be used for a wide variety of micro-machining applications including fabrication of microelectromechanical system (MEMS) components. Toward this end, a better understanding of the impacts the various machining parameters have on material removal has been established through a single discharge study of micro-EDM and a parametric study of small hole making by micro-EDM. The main avenues for improving the speed and efficiency of the micro-EDM process are in the areas of more controlled pulse generation in the power supply and more controlled positioning of the tool electrode during the machining process. Further investigation of the micro-EDM process in three dimensions leads to important design rules, specifically the smallest feature size attainable by the process.

  14. microRNAs in mycobacterial disease: friend or foe?

    Directory of Open Access Journals (Sweden)

    Manali D Mehta

    2014-07-01

    Full Text Available As the role of microRNA in all aspects of biology continues to be unraveled, the interplay between microRNAs and human disease is becoming clearer. It should come of no surprise that microRNAs play a major part in the outcome of infectious diseases, since early work has implicated microRNAs as regulators of the immune response. Here, we provide a review on how microRNAs influence the course of mycobacterial infections, which cause two of humanity’s most ancient infectious diseases: tuberculosis and leprosy. Evidence derived from profiling and functional experiments suggests that regulation of specific microRNAs during infection can either enhance the immune response or facilitate pathogen immune evasion. Now, it remains to be seen if the manipulation of host cell microRNA profiles can be an opportunity for therapeutic intervention for these difficult-to-treat diseases.

  15. Experimental and numerical study of micro deep drawing

    Directory of Open Access Journals (Sweden)

    Luo Liang

    2015-01-01

    Full Text Available Micro forming is a key technology for an industrial miniaturisation trend, and micro deep drawing (MDD is a typical micro forming method. It has great advantages comparing to other micro manufacturing methods, such as net forming ability, mass production potential, high product quality and complex 3D metal products fabrication capacity. Meanwhile, it is facing difficulties, for example the so-called size effects, once scaled down to micro scale. To investigate and to solve the problems in MDD, a combined micro blanking-drawing machine is employed and an explicit-implicit micro deep drawing model with a voronoi blank model is developed. Through heat treatment different grain sizes can be obtained, which affect material's properties and, consequently, the drawing process parameters, as well as produced cups' quality. Further, a voronoi model can provide detailed material information in simulation, and numerical simulation results are in accordance with experimental results.

  16. Dynamics of micro-bubble sonication inside a phantom vessel

    KAUST Repository

    Qamar, Adnan; Samtaney, Ravi; Bull, Joseph L.

    2013-01-01

    A model for sonicated micro-bubble oscillations inside a phantom vessel is proposed. The model is not a variant of conventional Rayleigh-Plesset equation and is obtained from reduced Navier-Stokes equations. The model relates the micro-bubble oscillation dynamics with geometric and acoustic parameters in a consistent manner. It predicts micro-bubble oscillation dynamics as well as micro-bubble fragmentation when compared to the experimental data. For large micro-bubble radius to vessel diameter ratios, predictions are damped, suggesting breakdown of inherent modeling assumptions for these cases. Micro-bubble response with acoustic parameters is consistent with experiments and provides physical insight to the micro-bubble oscillation dynamics.

  17. Dynamics of micro-bubble sonication inside a phantom vessel

    KAUST Repository

    Qamar, Adnan

    2013-01-10

    A model for sonicated micro-bubble oscillations inside a phantom vessel is proposed. The model is not a variant of conventional Rayleigh-Plesset equation and is obtained from reduced Navier-Stokes equations. The model relates the micro-bubble oscillation dynamics with geometric and acoustic parameters in a consistent manner. It predicts micro-bubble oscillation dynamics as well as micro-bubble fragmentation when compared to the experimental data. For large micro-bubble radius to vessel diameter ratios, predictions are damped, suggesting breakdown of inherent modeling assumptions for these cases. Micro-bubble response with acoustic parameters is consistent with experiments and provides physical insight to the micro-bubble oscillation dynamics.

  18. Fabrication of high aspect ratio micro electrode by using EDM

    International Nuclear Information System (INIS)

    Elsiti, Nagwa Mejid; Noordin, M.Y.; Alkali, Adam Umar

    2016-01-01

    The electrical discharge machining (EDM) process inherits characteristics that make it a promising micro-machining technique. Micro electrical discharge machining (micro- EDM) is a derived form of EDM, which is commonly used to manufacture micro and miniature parts and components by using the conventional electrical discharge machining fundamentals. Moving block electro discharge grinding (Moving BEDG) is one of the processes that can be used to fabricate micro-electrode. In this study, a conventional die sinker EDM machine was used to fabricate the micro-electrode. Modifications are made to the moving BEDG, which include changing the direction of movements and control gap in one electrode. Consequently current was controlled due to the use of roughing, semi-finishing and finishing parameters. Finally, a high aspect ratio micro-electrode with a diameter of 110.49μm and length of 6000μm was fabricated. (paper)

  19. Surface Micro Topography Replication in Injection Moulding

    DEFF Research Database (Denmark)

    Arlø, Uffe Rolf; Hansen, Hans Nørgaard; Kjær, Erik Michael

    2005-01-01

    The surface micro topography of injection moulded plastic parts can be important for aesthetical and technical reasons. The quality of replication of mould surface topography onto the plastic surface depends among other factors on the process conditions. A study of this relationship has been...... carried out with rough EDM (electrical discharge machining) mould surfaces, a PS grade, and by applying established three-dimensional topography parameters. Significant quantitative relationships between process parameters and topography parameters were established. It further appeared that replication...

  20. Computer Controlled Chemical Micro-Reactor

    International Nuclear Information System (INIS)

    Mechtilde, Schaefer; Eduard, Stach; Adreas, Foitzik

    2006-01-01

    Chemical reactions or chemical equilibria can be influenced and controlled by several parameters. The ratio of two liquid ingredients, the so called reactants or educts, plays an important role in determining the end product and its yield. The reactants must be weighed and accordingly mixed with the conventional batch mode. If the reaction is done in a microreactor or in several parallel working micro-reactors, units for allotting the educts in appropriate quantities are required. In this report we present a novel micro-reactor that allows the constant monitoring of the chemical reaction via Raman spectroscopy. Such monitoring enables an appropriate feedback on the steering parameters for the PC controlled micro-pumps for the appropriate educt flow rate of both liquids to get optimised ratios of ingredients at an optimised total flow rate. The micro-reactors are the core pieces of the design and are easily removable and can therefore be changed at any time to adapt the requirements of the chemical reaction. One type of reactor consists of a stainless steel base containing small scale milled channels covered with anodically bonded Pyrex glass. Another type of reactor has a base of anisotropically etched silicon, and is also covered with anodically bonded Pyrex glass. The glass window allows visual observation of the initial phase interface of the two educts in the reaction channels by optical microscopy and does not affect, in contrast to infrared spectroscopy, the Raman spectroscopic signal for detection of the reaction kinetics. On the basis of a test reaction, we present non-invasive and spatially highly resolved in-situ reaction analysis using Raman spectroscopy measured along the reaction channel at different locations