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Sample records for dystrophins carrying spectrin-like

  1. A Two-amino Acid Mutation Encountered in Duchenne Muscular Dystrophy Decreases Stability of the Rod Domain 23 (R23) Spectrin-like Repeat of Dystrophin.

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    Legardinier, Sébastien; Legrand, Baptiste; Raguénès-Nicol, Céline; Bondon, Arnaud; Hardy, Serge; Tascon, Christophe; Le Rumeur, Elisabeth; Hubert, Jean-François

    2009-03-27

    Lack of functional dystrophin causes severe Duchenne muscular dystrophy. The subsarcolemmal location of dystrophin, as well as its association with both cytoskeleton and membrane, suggests a role in the mechanical regulation of muscular membrane stress. In particular, phenotype rescue in a Duchenne muscular dystrophy mice model has shown that some parts of the central rod domain of dystrophin, constituted by 24 spectrin-like repeats, are essential. In this study, we made use of rare missense pathogenic mutations in the dystrophin gene and analyzed the biochemical properties of the isolated repeat 23 bearing single or double mutations E2910V and N2912D found in muscle dystrophy with severity grading. No dramatic effect on secondary and tertiary structure of the repeat was found in mutants compared with wild type as revealed by circular dichroism and NMR. Thermal and chemical unfolding data from circular dichroism and tryptophan fluorescence show significant decrease of stability for the mutants, and stopped-flow spectroscopy shows decreased refolding rates. The most deleterious single mutation is the N2912D replacement, although we observe additive effects of the two mutations on repeat stability. Based on three-dimensional structures built by homology molecular modeling, we discuss the modifications of the mutation-induced repeat stability. We conclude that the main forces involved in repeat stability are electrostatic inter-helix interactions that are disrupted following mutations. This study represents the first analysis at the protein level of the consequences of missense mutations in the human dystrophin rod domain. Our results suggest that it may participate in mechanical weakening of dystrophin-deficient muscle.

  2. Disruption of Spectrin-Like Cytoskeleton in Differentiating Keratinocytes by PKCδ Activation Is Associated with Phosphorylated Adducin

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    Zhao, Kong-Nan; Masci, Paul P.; Lavin, Martin F.

    2011-01-01

    Spectrin is a central component of the cytoskeletal protein network in a variety of erythroid and non-erythroid cells. In keratinocytes, this protein has been shown to be pericytoplasmic and plasma membrane associated, but its characteristics and function have not been established in these cells. Here we demonstrate that spectrin increases dramatically in amount and is assembled into the cytoskeleton during differentiation in mouse and human keratinocytes. The spectrin-like cytoskeleton was predominantly organized in the granular and cornified layers of the epidermis and disrupted by actin filament inhibitors, but not by anti-mitotic drugs. When the cytoskeleton was disrupted PKCδ was activated by phosphorylation on Thr505. Specific inhibition of PKCδ(Thr505) activation with rottlerin prevented disruption of the spectrin-like cytoskeleton and the associated morphological changes that accompany differentiation. Rottlerin also inhibited specific phosphorylation of the PKCδ substrate adducin, a cytoskeletal protein. Furthermore, knock-down of endogenous adducin affected not only expression of adducin, but also spectrin and PKCδ, and severely disrupted organization of the spectrin-like cytoskeleton and cytoskeletal distribution of both adducin and PKCδ. These results demonstrate that organization of a spectrin-like cytoskeleton is associated with keratinocytes differentiation, and disruption of this cytoskeleton is mediated by either PKCδ(Thr505) phosphorylation associated with phosphorylated adducin or due to reduction of endogenous adducin, which normally connects and stabilizes the spectrin-actin complex. PMID:22163289

  3. Disruption of spectrin-like cytoskeleton in differentiating keratinocytes by PKCδ activation is associated with phosphorylated adducin.

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    Kong-Nan Zhao

    Full Text Available Spectrin is a central component of the cytoskeletal protein network in a variety of erythroid and non-erythroid cells. In keratinocytes, this protein has been shown to be pericytoplasmic and plasma membrane associated, but its characteristics and function have not been established in these cells. Here we demonstrate that spectrin increases dramatically in amount and is assembled into the cytoskeleton during differentiation in mouse and human keratinocytes. The spectrin-like cytoskeleton was predominantly organized in the granular and cornified layers of the epidermis and disrupted by actin filament inhibitors, but not by anti-mitotic drugs. When the cytoskeleton was disrupted PKCδ was activated by phosphorylation on Thr505. Specific inhibition of PKCδ(Thr505 activation with rottlerin prevented disruption of the spectrin-like cytoskeleton and the associated morphological changes that accompany differentiation. Rottlerin also inhibited specific phosphorylation of the PKCδ substrate adducin, a cytoskeletal protein. Furthermore, knock-down of endogenous adducin affected not only expression of adducin, but also spectrin and PKCδ, and severely disrupted organization of the spectrin-like cytoskeleton and cytoskeletal distribution of both adducin and PKCδ. These results demonstrate that organization of a spectrin-like cytoskeleton is associated with keratinocytes differentiation, and disruption of this cytoskeleton is mediated by either PKCδ(Thr505 phosphorylation associated with phosphorylated adducin or due to reduction of endogenous adducin, which normally connects and stabilizes the spectrin-actin complex.

  4. Identification and function analysis of spectrin-like protein in pollen tubes of lily (Lilium davidii Duch)

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xueqin; YUAN Ming; WANG Xuechen

    2004-01-01

    The elongation of pollen tube is an importantprocess of sexual reproduction in higher plant. Cytoskeletonplays a major regulatory role in the elongation of pollentubes. But whether membrane skeleton is involved in thepollen tube elongation is not clear. In this study, immuno-chemical detection of spectrin-like protein has been carriedout in pollen tubes. By use of 2-dimensional electrophore-sis(2DE) and western blotting, two spectrin-like proteins arefound, one is 150 kD, and the other is 105 kD, with pI being4.54 and 4.39, respectively. 150 kD spectrin-like protein islocated in plasma membrane of pollen tube and 105 kDspectrin-like protein is located in cytoplasm, probably func-tioning as a subunit to form a dimmer (210 kD) in vivo. Theelongation of pollen tubes is inhibited after spectrin antibodywas injected into a growing pollen tube. These results suggestthat spectrin-like proteins exist in pollen tube and play animportant regulating role in the elongation process of pollentubes from lily.

  5. Chimeric snRNA molecules carrying antisense sequences against the splice junctions of exon 51 of the dystrophin pre-mRNA induce exon skipping and restoration of a dystrophin synthesis in Δ48-50 DMD cells

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    De Angelis, Fernanda Gabriella; Sthandier, Olga; Berarducci, Barbara; Toso, Silvia; Galluzzi, Giuliana; Ricci, Enzo; Cossu, Giulio; Bozzoni, Irene

    2002-01-01

    Deletions and point mutations in the dystrophin gene cause either the severe progressive myopathy Duchenne muscular dystrophy (DMD) or the milder Becker muscular dystrophy, depending on whether the translational reading frame is lost or maintained. Because internal in-frame deletions in the protein produce only mild myopathic symptoms, it should be possible, by preventing the inclusion of specific mutated exon(s) in the mature dystrophin mRNA, to restore a partially corrected phenotype. Such control has been previously accomplished by the use of synthetic oligonucleotides; nevertheless, a significant drawback to this approach is caused by the fact that oligonucleotides would require periodic administrations. To circumvent this problem, we have produced several constructs able to express in vivo, in a stable fashion, large amounts of chimeric RNAs containing antisense sequences. In this paper we show that antisense molecules against exon 51 splice junctions are able to direct skipping of this exon in the human DMD deletion 48–50 and to rescue dystrophin synthesis. We also show that the highest skipping activity was found when antisense constructs against the 5′ and 3′ splice sites are coexpressed in the same cell. PMID:12077324

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

    Science.gov (United States)

    Legrand, Baptiste; Giudice, Emmanuel; Nicolas, Aurélie; Delalande, Olivier; Le Rumeur, Elisabeth

    2011-01-01

    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.

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

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    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.

  8. Characterization of a 65 kDa NIF in the nuclear matrix of the monocot Allium cepa that interacts with nuclear spectrin-like proteins.

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    Pérez-Munive, Clara; Blumenthal, Sonal S D; de la Espina, Susana Moreno Díaz

    2012-01-01

    Plant cells have a well organized nucleus and nuclear matrix, but lack orthologues of the main structural components of the metazoan nuclear matrix. Although data is limited, most plant nuclear structural proteins are coiled-coil proteins, such as the NIFs (nuclear intermediate filaments) in Pisum sativum that cross-react with anti-intermediate filament and anti-lamin antibodies, form filaments 6-12 nm in diameter in vitro, and may play the role of lamins. We have investigated the conservation and features of NIFs in a monocot species, Allium cepa, and compared them with onion lamin-like proteins. Polyclonal antisera against the pea 65 kDa NIF were used in 1D and 2D Western blots, ICM (imunofluorescence confocal microscopy) and IEM (immunoelectron microscopy). Their presence in the nuclear matrix was analysed by differential extraction of nuclei, and their association with structural spectrin-like proteins by co-immunoprecipitation and co-localization in ICM. NIF is a conserved structural component of the nucleus and its matrix in monocots with Mr and pI values similar to those of pea 65 kDa NIF, which localized to the nuclear envelope, perichromatin domains and foci, and to the nuclear matrix, interacting directly with structural nuclear spectrin-like proteins. Its similarities with some of the proteins described as onion lamin-like proteins suggest that they are highly related or perhaps the same proteins.

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

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    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.

  10. 黄瓜韧皮部的类血影蛋白%Spectrin-like Protein in the Phloem of Cucumis sativus

    Institute of Scientific and Technical Information of China (English)

    邢立静; 花宝光; 娄成后

    2002-01-01

    Spectrin-like protein has been found in a variety of plant cells. In this study, electron microscopic observation of immuno-gold labelled preparations from the leaf petiole of cucumber (Cucumis sativus L.) shows that it also exists in the sieve element-companion cell (SE-CC) complex, being widely distributed in P-protein filaments and sieve element reticulum (SER), in the cytoplasm and mitochondrial membrane of companion cell (CC) and in the branched plasmodesmata between sieve element (SE) and CC as well. The results suggest that this protein could be synthesized in CC and transferred to SE through plasmodesmata. Western blotting showed that spectrin-like protein existed in the protein of phloem exudate of cucumber, and its molecular weight was about 260 kD.%以黄瓜(Cucumis sativus L.)叶柄为实验材料,应用胶体金免疫电镜技术证明类血影蛋白存在于韧皮部的筛管-伴胞复合体中,广泛分布于筛分子中的韧皮蛋白纤丝以及筛分子网络结构上,并且分布在伴胞的细胞质和线粒体膜以及筛分子与伴胞之间的分支状胞间连丝上,表明该蛋白可能由伴胞合成并经由二者之间的胞间连丝运输到筛分子中.用免疫印迹技术证明,黄瓜韧皮部汁液蛋白中存在类血影蛋白,其分子量约为260 kD,与动物细胞中血影蛋白的分子量接近.

  11. Carry

    DEFF Research Database (Denmark)

    Koijen, Ralph S.J.; Moskowitz, Tobias J.; Heje Pedersen, Lasse;

    that include global equities, global bonds, currencies, commodities, US Treasuries, credit, and equity index options. This predictability underlies the strong returns to "carry trades" that go long high-carry and short low-carry securities, applied almost exclusively to currencies, but shown here...

  12. Efficient Restoration of the Dystrophin Gene Reading Frame and Protein Structure in DMD Myoblasts Using the CinDel Method

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    Jean-Paul Iyombe-Engembe

    2016-01-01

    Full Text Available The CRISPR/Cas9 system is a great revolution in biology. This technology allows the modification of genes in vitro and in vivo in a wide variety of living organisms. In most Duchenne muscular dystrophy (DMD patients, expression of dystrophin (DYS protein is disrupted because exon deletions result in a frame shift. We present here the CRISPR-induced deletion (CinDel, a new promising genome-editing technology to correct the DMD gene. This strategy is based on the use of two gRNAs targeting specifically exons that precede and follow the patient deletion in the DMD gene. This pair of gRNAs induced a precise large additional deletion leading to fusion of the targeted exons. Using an adequate pair of gRNAs, the deletion of parts of these exons and the intron separating them restored the DMD reading frame in 62% of the hybrid exons in vitro in DMD myoblasts and in vivo in electroporated hDMD/mdx mice. Moreover, adequate pairs of gRNAs also restored the normal spectrin-like repeat of the dystrophin rod domain; such restoration is not obtained by exon skipping or deletion of complete exons. The expression of an internally deleted DYS protein was detected following the formation of myotubes by the unselected, treated DMD myoblasts. Given that CinDel induces permanent reparation of the DMD gene, this treatment would not have to be repeated as it is the case for exon skipping induced by oligonucleotides.

  13. Dystrophin-Deficient Cardiomyopathy.

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    Kamdar, Forum; Garry, Daniel J

    2016-05-31

    Dystrophinopathies are a group of distinct neuromuscular diseases that result from mutations in the structural cytoskeletal Dystrophin gene. Dystrophinopathies include Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD), X-linked dilated cardiomyopathy, as well as DMD and BMD female carriers. The primary presenting symptom in most dystrophinopathies is skeletal muscle weakness. However, cardiac muscle is also a subtype of striated muscle and is similarly affected in many of the muscular dystrophies. Cardiomyopathies associated with dystrophinopathies are an increasingly recognized manifestation of these neuromuscular disorders and contribute significantly to their morbidity and mortality. Recent studies suggest that these patient populations would benefit from cardiovascular therapies, annual cardiovascular imaging studies, and close follow-up with cardiovascular specialists. Moreover, patients with DMD and BMD who develop end-stage heart failure may benefit from the use of advanced therapies. This review focuses on the pathophysiology, cardiac involvement, and treatment of cardiomyopathy in the dystrophic patient. Copyright © 2016 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.

  14. Electrotransfer of the full-length dog dystrophin into mouse and dystrophic dog muscles.

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    Pichavant, Christophe; Chapdelaine, Pierre; Cerri, Daniel G; Bizario, Joao C S; Tremblay, Jacques P

    2010-11-01

    Duchenne muscular dystrophy (DMD) is an X-linked genetic disease characterized by the absence of dystrophin (427 kDa). An approach to eventually restore this protein in patients with DMD is to introduce into their muscles a plasmid encoding dystrophin cDNA. Because the phenotype of the dystrophic dog is closer to the human phenotype than is the mdx mouse phenotype, we have studied the electrotransfer of a plasmid carrying the full-length dog dystrophin (FLDYS(dog)) in dystrophic dog muscle. To achieve this nonviral delivery, the FLDYS(dog) cDNA was cloned in two plasmids containing either a cytomegalovirus or a muscle creatine kinase promoter. In both cases, our results showed that the electrotransfer of these large plasmids (∼17 kb) into mouse muscle allowed FLDYS(dog) expression in the treated muscle. The electrotransfer of pCMV.FLDYS(dog) in a dystrophic dog muscle also led to the expression of dystrophin. In conclusion, introduction of the full-length dog dystrophin cDNA by electrotransfer into dystrophic dog muscle is a potential approach to restore dystrophin in patients with DMD. However, the electrotransfer procedure should be improved before applying it to humans.

  15. DMD transcript imbalance determines dystrophin levels.

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    Spitali, Pietro; van den Bergen, Janneke C; Verhaart, Ingrid E C; Wokke, Beatrijs; Janson, Anneke A M; van den Eijnde, Rani; den Dunnen, Johan T; Laros, Jeroen F J; Verschuuren, Jan J G M; 't Hoen, Peter A C; Aartsma-Rus, Annemieke

    2013-12-01

    Duchenne and Becker muscular dystrophies are caused by out-of-frame and in-frame mutations, respectively, in the dystrophin encoding DMD gene. Molecular therapies targeting the precursor-mRNA are in clinical trials and show promising results. These approaches will depend on the stability and expression levels of dystrophin mRNA in skeletal muscles and heart. We report that the DMD gene is more highly expressed in heart than in skeletal muscles, in mice and humans. The transcript mutated in the mdx mouse model shows a 5' to 3' imbalance compared with that of its wild-type counterpart and reading frame restoration via antisense-mediated exon skipping does not correct this event. We also report significant transcript instability in 22 patients with Becker dystrophy, clarifying the fact that transcript imbalance is not caused by premature nonsense mutations. Finally, we demonstrate that transcript stability, rather than transcriptional rate, is an important determinant of dystrophin protein levels in patients with Becker dystrophy. We suggest that the availability of the complete transcript is a key factor to determine protein abundance and thus will influence the outcome of mRNA-targeting therapies.

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

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    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…

  17. Dystrophin insufficiency causes selective muscle histopathology and loss of dystrophin-glycoprotein complex assembly in pig skeletal muscle

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    Duchenne muscular dystrophy (DMD) is caused by a dystrophin deficiency while Becker muscular dystrophy (BMD) is caused by a dystrophin insufficiency or expression of a partially functional protein product. Both of these dystrophinopathies are most commonly studied using the mdx mouse and a golden r...

  18. Expression of dystrophin-glycoprotein complex at the skeletal muscle sarcolemma in Duchenne muscular dystrophy

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    Lei ZHAO

    2015-07-01

    Full Text Available Background  Eccentric exercise or high tension exercise could cause damage to skeletal muscle structure, resulting in deficiency of dystrophin and secondary loss of dystrophin-glycoprotein complex (DGC from the sarcolemma, which indicated that down-regulation of dystrophin was one of the key points of skeletal muscle injury from eccentric exercise. Duchenne muscular dystrophy (DMD is caused by mutations of DMD gene, resulting in the absence of dystrophin, which means that skeletal muscles of DMD patients after birth are in the natural state of actual path of force transmission which carried high tension from eccentric exercise. This paper investigated systematically whether expression of DGC is associated with progressive muscle weakness in natural history of DMD, and analyzed the expression of DGC at the sarcolemma of 197 confirmed DMD cases (9 days-12 years old.  Methods  The expression of α- and β-dystroglycan (DG, α-, β-, γ- and δ-sarcoglycan (SG and syntrophin at the sarcolemma of DMD patients was analyzed by immunofluorescent staining.  Results  The study showed that there was no relationship between lack of proteins and progressive muscle weakness with increasing age, although expression of α- and β-DG, α-, β-, γ- and δ-SG and syntrophin at the sarcolemma at different stages of 197 DMD patients (9 days-12 years old had different degrees of deficiency.  Conclusions  Deficiency of DGC may occur before birth and DMD patients were recommended to avoid further damage to skeletal muscles from eccentric exercise and high-resistance movement in activities of daily life and rehabilitation training. DOI: 10.3969/j.issn.1672-6731.2015.06.006

  19. A marginal level of dystrophin partially ameliorates hindlimb muscle passive mechanical properties in dystrophin-null mice.

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    Hakim, Chady H; Duan, Dongsheng

    2012-12-01

    The goal of this study was to determine whether a minimal level of dystrophin expression improves the passive mechanical properties of skeletal muscle in the murine Duchenne muscular dystrophy model. We compared the elastic and viscous properties of the extensor digitorum longus muscle (EDL) in mdx3cv and mdx4cv mice at 6, 14, and 20 months of age. Both strains are on the C57Bl/6 background, and both lose the full-length dystrophin protein. Interestingly, mdx3cv mice express a near full-length dystrophin at ≈ 5% of the normal level. We found that the stress-strain profile and the stress relaxation rate of the EDL in mdx3cv mice were partially preserved in all age groups compared with age-matched mdx4cv mice. Our results suggest that a low level of dystrophin expression may treat muscle stiffness in Duchenne muscular dystrophy. Copyright © 2012 Wiley Periodicals, Inc.

  20. Becker muscular dystrophy in Indian patients: Analysis of dystrophin gene deletion patterns

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    Dastur Rashna

    2008-01-01

    Full Text Available Background: Becker muscular dystrophy (BMD is caused by mutations in the dystrophin gene with variable phenotypes. Becker muscular dystrophy patients have low levels of nearly full-length dystrophin and carry in-frame mutations, which allow partial functioning of the protein. Aim: To study the deletion patterns of BMD and to correlate the same with reading frame rule and different phenotypes. Setting: A tertiary care teaching hospital. Design: This is a prospective hospital-based study. Materials and Methods: Thirty-two exons spanning different "hot spot" regions using Multiplex PCR techniques were studied in 347 patients. Two hundred and twenty-two showed deletions in one or more of the 32 exons. Out of these, 46 diagnosed as BMD patients were analyzed. Results: Forty-six BMD patients showed deletions in both regions of the dystrophin gene. Out of these 89.1% (41/46 were in-frame deletions. Deletions starting with Exon 45 were found in 76.1% (35/46 of the cases. Mutations in the majority of cases i.e. 39/46 (84.8% were seen in 3′ downstream region (Exon 45-55, distal rod domain. Few, i.e. 5/46 (10.8% showed deletions in 5′ upstream region (Exons 3-20, N-terminus and proximal rod domain of the gene, while in 2/46 (4.4% large mutations (>40 bp spanning both regions (Exons 3-55 were detected. Conclusion: This significant gene deletion analysis has been carried out for BMD patients particularly from Western India using 32 exons.

  1. Accurate Quantitation of Dystrophin Protein in Human Skeletal Muscle Using Mass Spectrometry

    OpenAIRE

    Brown, Kristy J; Marathi, Ramya; Fiorillo, Alyson A; Ciccimaro, Eugene F.; Sharma, Seema; Rowlands, David S.; Rayavarapu, Sree; Nagaraju, Kanneboyina; Eric P. Hoffman; Hathout, Yetrib

    2012-01-01

    Quantitation of human dystrophin protein in muscle biopsies is a clinically relevant endpoint for both diagnosis and response to dystrophin-replacement therapies for dystrophinopathies. A robust and accurate assay would enable the use of dystrophin as a surrogate biomarker, particularly in exploratory Phase 2 trials. Currently available methods to quantitate dystrophin rely on immunoblot or immunohistochemistry methods that are not considered robust. Here we present a mass spectrometry based ...

  2. Expression of dystrophin-glycoprotein complex at the skeletal muscle sarcolemma in Duchenne muscular dystrophy

    OpenAIRE

    Zhao, Lei; Chao-ping HU; Wang, Yi; Shui-zhen ZHOU; Shi, Yi-Yun; Xi-hua LI

    2015-01-01

    Background  Eccentric exercise or high tension exercise could cause damage to skeletal muscle structure, resulting in deficiency of dystrophin and secondary loss of dystrophin-glycoprotein complex (DGC) from the sarcolemma, which indicated that down-regulation of dystrophin was one of the key points of skeletal muscle injury from eccentric exercise. Duchenne muscular dystrophy (DMD) is caused by mutations of DMD gene, resulting in the absence of dystrophin, which means that skeletal muscles o...

  3. Dystrophin and utrophin influence fiber type composition and post-synaptic membrane structure.

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    Rafael, J A; Townsend, E R; Squire, S E; Potter, A C; Chamberlain, J S; Davies, K E

    2000-05-22

    The X-linked muscle wasting disease Duchenne muscular dystrophy is caused by the lack of dystrophin in muscle. Protein structure predictions, patient mutations, in vitro binding studies and transgenic and knockout mice suggest that dystrophin plays a mechanical role in skeletal muscle, linking the subsarcolemmal cytoskeleton with the extracellular matrix through its direct interaction with the dystrophin-associated protein complex (DAPC). Although a signaling role for dystrophin has been postulated, definitive data have been lacking. To identify potential non-mechanical roles of dystrophin, we tested the ability of various truncated dystrophin transgenes to prevent any of the skeletal muscle abnormalities associated with the double knockout mouse deficient for both dystrophin and the dystrophin-related protein utrophin. We show that restoration of the DAPC with Dp71 does not prevent the structural abnormalities of the post-synaptic membrane or the abnormal oxidative properties of utrophin/dystrophin-deficient muscle. In marked contrast, a dystrophin protein lacking the cysteine-rich domain, which is unable to prevent dystrophy in the mdx mouse, is able to ameliorate these abnormalities in utrophin/dystrophin-deficient mice. These experiments provide the first direct evidence that in addition to a mechanical role and relocalization of the DAPC, dystrophin and utrophin are able to alter both structural and biochemical properties of skeletal muscle. In addition, these mice provide unique insights into skeletal muscle fiber type composition.

  4. The roles of dystrophin and dystrobrevin : in synaptic signaling in drosophila

    NARCIS (Netherlands)

    Potikanond, Saranyapin

    2012-01-01

    Duchenne muscular dystrophy (DMD) is a disease, characterized by progressive muscle wasting, caused by the lack of Dystrophin. A subset of DMD patients also have cognitive deficits likely due to the absence of Dystrophin from brain synapses where it is usually localized. Dystrophin and a number of o

  5. Transcriptomic analysis of dystrophin RNAi knockdown reveals a central role for dystrophin in muscle differentiation and contractile apparatus organization

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    Graham Ian R

    2010-06-01

    Full Text Available Abstract Background Duchenne muscular dystrophy (DMD is a fatal muscle wasting disorder caused by mutations in the dystrophin gene. DMD has a complex and as yet incompletely defined molecular pathophysiology hindering development of effective ameliorative approaches. Transcriptomic studies so far conducted on dystrophic cells and tissues suffer from non-specific changes and background noise due to heterogeneous comparisons and secondary pathologies. A study design in which a perfectly matched control cell population is used as reference for transcriptomic studies will give a much more specific insight into the effects of dystrophin deficiency and DMD pathophysiology. Results Using RNA interference (RNAi to knock down dystrophin in myotubes from C57BL10 mice, we created a homogenous model to study the transcriptome of dystrophin-deficient myotubes. We noted significant differences in the global gene expression pattern between these myotubes and their matched control cultures. In particular, categorical analyses of the dysregulated genes demonstrated significant enrichment of molecules associated with the components of muscle cell contractile unit, ion channels, metabolic pathways and kinases. Additionally, some of the dysregulated genes could potentially explain conditions and endophenotypes associated with dystrophin deficiency, such as dysregulation of calcium homeostasis (Pvalb and Casq1, or cardiomyopathy (Obscurin, Tcap. In addition to be validated by qPCR, our data gains another level of validity by affirmatively reproducing several independent studies conducted previously at genes and/or protein levels in vivo and in vitro. Conclusion Our results suggest that in striated muscles, dystrophin is involved in orchestrating proper development and organization of myofibers as contractile units, depicting a novel pathophysiology for DMD where the absence of dystrophin results in maldeveloped myofibers prone to physical stress and damage

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

    Directory of Open Access Journals (Sweden)

    Dejia Li

    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.

  7. Disodium cromoglycate protects dystrophin-deficient muscle fibers from leakiness.

    Science.gov (United States)

    Marques, Maria Julia; Ventura Machado, Rafael; Minatel, Elaine; Santo Neto, Humberto

    2008-01-01

    In dystrophin-deficient fibers of mdx mice and in Duchenne dystrophy, the lack of dystrophin leads to sarcolemma breakdown and muscle degeneration. We verified that cromolyn, a mast-cell stabilizer agent, stabilized dystrophic muscle fibers using Evans blue dye as a marker of sarcolemma leakiness. Mdx mice (n=8; 14 days of age) received daily intraperitoneal injections of cromolyn (50 mg/kg body weight) for 15 days. Untreated mdx mice (n=8) were injected with saline. Cryostat cross-sections of the sternomastoid, tibialis anterior, and diaphragm muscles were stained with hematoxylin and eosin. Cromolyn dramatically reduced Evans blue dye-positive fibers in all muscles (P<0.05; Student's t-test) and led to a significant increase in the percentage of fibers with peripheral nuclei. This study supports the protective effects of cromolyn in dystrophic muscles and further indicates its action against muscle fiber leakiness in muscles that are differently affected by the lack of dystrophin.

  8. Dystrophin, utrophin and {beta}-dystroglycan expression in skeletal muscle from patients with Becker muscular dystrophy

    Energy Technology Data Exchange (ETDEWEB)

    Kawajiri, Masakazu; Mitsui, Takao; Kawai, Hisaomi [Univ. of Tokushima (Japan)] [and others

    1996-08-01

    The precise localization and semiquantitative correlation of dystrophin, utrophin and {beta}-dystroglycan expression on the sarcolemma of skeletal muscle cells obtained from patients with Becker muscular dystrophy (BMD) was studied using three types of double immunofluorescence. Staining intensity was measured using a confocal laser microscope. Each of these proteins was identified at the same locus on the sarcolemma. The staining intensities of dystrophin and utrophin were approximately reciprocal at sarcolemmal sites where dystrophin expression was obviously observed. The staining intensity of {beta}-dystroglycan was strong in areas where dystrophin staining was also strong and utrophin expression was weak. Quantitative analysis revealed that the staining intensity of {beta}-dystroglycan minus that of dystrophin approximated the staining intensity of utrophin, indicating that the sum of dystrophin and utrophin expression corresponds to that of {beta}-dystroglycan. These results suggest that utrophin may compensate for dystrophin deficiency found in BMD by binding to {beta}-dystroglycan. 35 refs., 3 figs., 1 tab.

  9. The dystrophin gene and cognitive function in the general population

    NARCIS (Netherlands)

    D. Vojinovic (Dina); H.H.H. Adams (Hieab); S. van der Lee (Sven); C.A. Ibrahim-Verbaas (Carla); R.W.W. Brouwer (Rutger); M.C.G.N. van den hout (Mirjam); E. Oole (Edwin); J. van Rooij (Jeroen); A.G. Uitterlinden (André); A. Hofman (Albert); W.F.J. van IJcken (Wilfred); A. Aartsma-Rus (Annemieke); G.-J.B. Van Ommen (Gert-Jan B.); M.A. Ikram (Arfan); C.M. van Duijn (Cornelia M.); N. Amin (Najaf)

    2015-01-01

    textabstractThe aim of our study is to investigate whether single-nucleotide dystrophin gene (DMD) variants associate with variability in cognitive functions in healthy populations. The study included 1240 participants from the Erasmus Rucphen family (ERF) study and 1464 individuals from the Rotterd

  10. Role of dystrophins and utrophins in platelet adhesion process.

    Science.gov (United States)

    Cerecedo, Doris; Mondragón, Ricardo; Cisneros, Bulmaro; Martínez-Pérez, Francisco; Martínez-Rojas, Dalila; Rendón, Alvaro

    2006-07-01

    Platelets are crucial at the site of vascular injury, adhering to the sub-endothelial matrix through receptors on their surface, leading to cell activation and aggregation to form a haemostatic plug. Platelets display focal adhesions as well as stress fibres to contract and facilitate expulsion of growth and pro-coagulant factors contained in the granules and to constrict the clot. The interaction of F-actin with different actin-binding proteins determines the properties and composition of the focal adhesions. Recently, we demonstrated the presence of dystrophin-associated protein complex corresponding to short dystrophin isoforms (Dp71d and Dp71) and the uthophin gene family (Up400 and Up71), which promote shape change, adhesion, aggregation, and granule centralisation. To elucidate participation of both complexes during the platelet adhesion process, their potential association with integrin beta-1 fraction and the focal adhesion system (alpha-actinin, vinculin and talin) was evaluated by immunofluorescence and immunoprecipitation assays. It was shown that the short dystrophin-associated protein complex participated in stress fibre assembly and in centralisation of cytoplasmic granules, while the utrophin-associated protein complex assembled and regulated focal adhesions. The simultaneous presence of dystrophin and utrophin complexes indicates complementary structural and signalling mechanisms to the actin network, improving the platelet haemostatic role.

  11. Muscle dysfunction and structural defects of dystrophin-null sapje mutant zebrafish larvae are rescued by ataluren treatment.

    Science.gov (United States)

    Li, Mei; Andersson-Lendahl, Monika; Sejersen, Thomas; Arner, Anders

    2014-04-01

    Sapje zebrafish carry a mutation in the dystrophin gene, which results in a premature stop codon, and a severe muscle phenotype. They display several of the structural characteristics of Duchenne muscular dystrophy (DMD). Ataluren (PTC124) is proposed to cause readthrough of premature stop codons and has been introduced as a potential treatment of genetic disorders. Clinical trials in DMD have shown promise, although with complex dose dependency. We have established physiology techniques, enabling high resolution of contractile function in skeletal muscle of zebrafish larvae. We aimed to provide a mechanical analysis of sapje larval muscle and examine effects of ataluren. Homozygous 5 d postfertilization (dpf) sapje larvae exhibited structural defects with 50% decrease in active tension. Ataluren (0.1-1 μM, 3-5 dpf) improved contractile function (~60% improvement of force at 0.5 μM) and dystrophin expression. Controls were not affected. Higher doses (5 μM, 35 μM) impaired contractile function, an effect also observed in controls, suggesting unspecific negative effects at high concentrations. In summary, Sapje larvae exhibit impaired contractile performance and provide a relevant DMD model for functional studies. Ataluren significantly improves skeletal muscle function in the sapje larvae, most likely reflecting an observed increase in dystrophin expression. The bell-shaped dose dependence in sapje resembles that previously reported in clinical DMD studies.

  12. Dystrophins, Utrophins, and Associated Scaffolding Complexes: Role in Mammalian Brain and Implications for Therapeutic Strategies

    Directory of Open Access Journals (Sweden)

    Caroline Perronnet

    2010-01-01

    Full Text Available Two decades of molecular, cellular, and functional studies considerably increased our understanding of dystrophins function and unveiled the complex etiology of the cognitive deficits in Duchenne muscular dystrophy (DMD, which involves altered expression of several dystrophin-gene products in brain. Dystrophins are normally part of critical cytoskeleton-associated membrane-bound molecular scaffolds involved in the clustering of receptors, ion channels, and signaling proteins that contribute to synapse physiology and blood-brain barrier function. The utrophin gene also drives brain expression of several paralogs proteins, which cellular expression and biological roles remain to be elucidated. Here we review the structural and functional properties of dystrophins and utrophins in brain, the consequences of dystrophins loss-of-function as revealed by numerous studies in mouse models of DMD, and we discuss future challenges and putative therapeutic strategies that may compensate for the cognitive impairment in DMD based on experimental manipulation of dystrophins and/or utrophins brain expression.

  13. Carrying Capacity

    DEFF Research Database (Denmark)

    Schroll, Henning; Andersen, Jan; Kjærgård, Bente

    2012-01-01

    A spatial planning act was introduced inIndonesia 1992 and renewed in 2008. It emphasised the planning role of decentralised authorities. The spatial planning act covers both spatial and environmental issues. It defines the concept of carrying capacity and includes definitions of supportive...... carrying capacity (SCC) and assimilative carrying capacity (ACC). The act mandates that the latter two aspects must be taken into consideration in the local spatial plans. The present study aimed at developing a background for a national guideline for carrying capacity in Indonesian provinces and districts...... standard or governmental political objective exists. In most cases it was possible to select a set of indicators, including thresholds that are workable in a carrying capacity planning at the local administrative levels. Not all relevant sectors at the decentralized level were included. Indicators of SCC...

  14. Dystrophin Distribution and Expression in Human and Experimental Temporal Lobe Epilepsy

    Science.gov (United States)

    Hendriksen, Ruben G. F.; Schipper, Sandra; Hoogland, Govert; Schijns, Olaf E. M. G.; Dings, Jim T. A.; Aalbers, Marlien W.; Vles, Johan S. H.

    2016-01-01

    Objective: Dystrophin is part of a protein complex that connects the cytoskeleton to the extracellular matrix. In addition to its role in muscle tissue, it functions as an anchoring protein within the central nervous system such as in hippocampus and cerebellum. Its presence in the latter regions is illustrated by the cognitive problems seen in Duchenne Muscular Dystrophy (DMD). Since epilepsy is also supposed to constitute a comorbidity of DMD, it is hypothesized that dystrophin plays a role in neuronal excitability. Here, we aimed to study brain dystrophin distribution and expression in both, human and experimental temporal lobe epilepsy (TLE). Method: Regional and cellular dystrophin distribution was evaluated in both human and rat hippocampi and in rat cerebellar tissue by immunofluorescent colocalization with neuronal (NeuN and calbindin) and glial (GFAP) markers. In addition, hippocampal dystrophin levels were estimated by Western blot analysis in biopsies from TLE patients, post-mortem controls, amygdala kindled (AK)-, and control rats. Results: Dystrophin was expressed in all hippocampal pyramidal subfields and in the molecular-, Purkinje-, and granular cell layer of the cerebellum. In these regions it colocalized with GFAP, suggesting expression in astrocytes such as Bergmann glia (BG) and velate protoplasmic astrocytes. In rat hippocampus and cerebellum there were neither differences in dystrophin positive cell types, nor in the regional dystrophin distribution between AK and control animals. Quantitatively, hippocampal full-length dystrophin (Dp427) levels were about 60% higher in human TLE patients than in post-mortem controls (p < 0.05), whereas the level of the shorter Dp71 isoform did not differ. In contrast, AK animals showed similar dystrophin levels as controls. Conclusion: Dystrophin is ubiquitously expressed by astrocytes in the human and rat hippocampus and in the rat cerebellum. Hippocampal full-length dystrophin (Dp427) levels are upregulated

  15. Carrying Capacity

    DEFF Research Database (Denmark)

    Schroll, Henning; Andersen, Jan; Kjærgård, Bente

    2012-01-01

    A spatial planning act was introduced inIndonesia 1992 and renewed in 2008. It emphasised the planning role of decentralised authorities. The spatial planning act covers both spatial and environmental issues. It defines the concept of carrying capacity and includes definitions of supportive...... and ACC may increase the political focus on resources and environmental issues and may help to move local authorities towards a more holistic spatial planning approach. A carrying capacity approach could be an inspiration for local spatial planning in developing countries. A spatial planning act...... was introduced inIndonesia 1992 and renewed in 2008. It emphasised the planning role of decentralised authorities. The spatial planning act covers both spatial and environmental issues. It defines the concept of carrying capacity and includes definitions of supportive carrying capacity (SCC) and assimilative...

  16. Detection of an exon 53 polymorphism in the dystrophin gene.

    Science.gov (United States)

    Prior, T W; Papp, A C; Snyder, P J; Sedra, M S

    1993-10-01

    We utilized a heteroduplex method to screen for small mutations in Duchenne muscular dystrophy patients who did not have deletions or duplications. A dystrophin exon 53 heteroduplex band was identified in 14.4% of the affected patients. Direct sequencing of the amplified product from DNA producing the heteroduplex revealed the presence of a polymorphism in the coding region. The codon for asparagine was converted from AAT to AAC.

  17. 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.

  18. 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.

  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.

    Science.gov (United States)

    Prior, T W; Bartolo, C; Pearl, D K; Papp, A C; Snyder, P J; Sedra, M S; Burghes, A H; Mendell, J R

    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.

  1. Autologous skeletal muscle derived cells expressing a novel functional dystrophin provide a potential therapy for Duchenne Muscular Dystrophy.

    Science.gov (United States)

    Meng, Jinhong; Counsell, John R; Reza, Mojgan; Laval, Steven H; Danos, Olivier; Thrasher, Adrian; Lochmüller, Hanns; Muntoni, Francesco; Morgan, Jennifer E

    2016-01-27

    Autologous stem cells that have been genetically modified to express dystrophin are a possible means of treating Duchenne Muscular Dystrophy (DMD). To maximize the therapeutic effect, dystrophin construct needs to contain as many functional motifs as possible, within the packaging capacity of the viral vector. Existing dystrophin constructs used for transduction of muscle stem cells do not contain the nNOS binding site, an important functional motif within the dystrophin gene. In this proof-of-concept study, using stem cells derived from skeletal muscle of a DMD patient (mdcs) transplanted into an immunodeficient mouse model of DMD, we report that two novel dystrophin constructs, C1 (ΔR3-R13) and C2 (ΔH2-R23), can be lentivirally transduced into mdcs and produce dystrophin. These dystrophin proteins were functional in vivo, as members of the dystrophin glycoprotein complex were restored in muscle fibres containing donor-derived dystrophin. In muscle fibres derived from cells that had been transduced with construct C1, the largest dystrophin construct packaged into a lentiviral system, nNOS was restored. The combination of autologous stem cells and a lentivirus expressing a novel dystrophin construct which optimally restores proteins of the dystrophin glycoprotein complex may have therapeutic application for all DMD patients, regardless of their dystrophin mutation.

  2. 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 de

  3. 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.

  4. Overexpression of dystrophin in transgenic mdx mice eliminates dystrophic symptoms without toxicity.

    Science.gov (United States)

    Cox, G A; Cole, N M; Matsumura, K; Phelps, S F; Hauschka, S D; Campbell, K P; Faulkner, J A; Chamberlain, J S

    1993-08-19

    Duchenne and Becker muscular dystrophy (DMD and BMD) are X-linked recessive diseases caused by defective expression of dystrophin. The mdx mouse, an animal model for DMD, has a mutation that eliminates expression of the 427K muscle and brain isoforms of dystrophin. Although these animals do not display overt muscle weakness or impaired movement, the diaphragm muscle of the mdx mouse is severely affected and shows progressive myofibre degeneration and fibrosis which closely resembles the human disease. Here we explore the feasibility of gene therapy for DMD by examining the potential of a full-length dystrophin transgene to correct dystrophic symptoms in mdx mice. We find that expression of dystrophin in muscles of transgenic mdx mice eliminates the morphological and immunohistological symptoms of muscular dystrophy. In addition, overexpression of dystrophin prevents the development of the abnormal mechanical properties associated with dystrophic muscle without causing deleterious side effects. Our results provide functional evidence for the feasibility of gene therapy for DMD.

  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. Novel Nuclear Protein Complexes of Dystrophin 71 Isoforms in Rat Cultured Hippocampal GABAergic and Glutamatergic Neurons.

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    Rafael Rodríguez-Muñoz

    Full Text Available The precise functional role of the dystrophin 71 in neurons is still elusive. Previously, we reported that dystrophin 71d and dystrophin 71f are present in nuclei from cultured neurons. In the present work, we performed a detailed analysis of the intranuclear distribution of dystrophin 71 isoforms (Dp71d and Dp71f, during the temporal course of 7-day postnatal rats hippocampal neurons culture for 1h, 2, 4, 10, 15 and 21 days in vitro (DIV. By immunofluorescence assays, we detected the highest level of nuclear expression of both dystrophin Dp71 isoforms at 10 DIV, during the temporal course of primary culture. Dp71d and Dp71f were detected mainly in bipolar GABAergic (≥60% and multipolar Glutamatergic (≤40% neurons, respectively. We also characterized the existence of two nuclear dystrophin-associated protein complexes (DAPC: dystrophin 71d or dystrophin 71f bound to β-dystroglycan, α1-, β-, α2-dystrobrevins, α-syntrophin, and syntrophin-associated protein nNOS (Dp71d-DAPC or Dp71f-DAPC, respectively, in the hippocampal neurons. Furthermore, both complexes were localized in interchromatin granule cluster structures (nuclear speckles of neuronal nucleoskeleton preparations. The present study evinces that each Dp71's complexes differ slightly in dystrobrevins composition. The results demonstrated that Dp71d-DAPC was mainly localized in bipolar GABAergic and Dp71f-DAPC in multipolar Glutamatergic hippocampal neurons. Taken together, our results show that dystrophin 71d, dystrophin 71f and DAP integrate protein complexes, and both complexes were associated to nuclear speckles structures.

  7. Novel Nuclear Protein Complexes of Dystrophin 71 Isoforms in Rat Cultured Hippocampal GABAergic and Glutamatergic Neurons.

    Science.gov (United States)

    Rodríguez-Muñoz, Rafael; Cárdenas-Aguayo, María Del Carmen; Alemán, Víctor; Osorio, Beatriz; Chávez-González, Oscar; Rendon, Alvaro; Martínez-Rojas, Dalila; Meraz-Ríos, Marco Antonio

    2015-01-01

    The precise functional role of the dystrophin 71 in neurons is still elusive. Previously, we reported that dystrophin 71d and dystrophin 71f are present in nuclei from cultured neurons. In the present work, we performed a detailed analysis of the intranuclear distribution of dystrophin 71 isoforms (Dp71d and Dp71f), during the temporal course of 7-day postnatal rats hippocampal neurons culture for 1h, 2, 4, 10, 15 and 21 days in vitro (DIV). By immunofluorescence assays, we detected the highest level of nuclear expression of both dystrophin Dp71 isoforms at 10 DIV, during the temporal course of primary culture. Dp71d and Dp71f were detected mainly in bipolar GABAergic (≥60%) and multipolar Glutamatergic (≤40%) neurons, respectively. We also characterized the existence of two nuclear dystrophin-associated protein complexes (DAPC): dystrophin 71d or dystrophin 71f bound to β-dystroglycan, α1-, β-, α2-dystrobrevins, α-syntrophin, and syntrophin-associated protein nNOS (Dp71d-DAPC or Dp71f-DAPC, respectively), in the hippocampal neurons. Furthermore, both complexes were localized in interchromatin granule cluster structures (nuclear speckles) of neuronal nucleoskeleton preparations. The present study evinces that each Dp71's complexes differ slightly in dystrobrevins composition. The results demonstrated that Dp71d-DAPC was mainly localized in bipolar GABAergic and Dp71f-DAPC in multipolar Glutamatergic hippocampal neurons. Taken together, our results show that dystrophin 71d, dystrophin 71f and DAP integrate protein complexes, and both complexes were associated to nuclear speckles structures.

  8. Proteomic Profiling of the Dystrophin-Deficient MDX Heart Reveals Drastically Altered Levels of Key Metabolic and Contractile Proteins

    Directory of Open Access Journals (Sweden)

    Caroline Lewis

    2010-01-01

    Full Text Available Although Duchenne muscular dystrophy is primarily classified as a neuromuscular disease, cardiac complications play an important role in the course of this X-linked inherited disorder. The pathobiochemical steps causing a progressive decline in the dystrophic heart are not well understood. We therefore carried out a fluorescence difference in-gel electrophoretic analysis of 9-month-old dystrophin-deficient versus age-matched normal heart, using the established MDX mouse model of muscular dystrophy-related cardiomyopathy. Out of 2,509 detectable protein spots, 79 2D-spots showed a drastic differential expression pattern, with the concentration of 3 proteins being increased, including nucleoside diphosphate kinase and lamin-A/C, and of 26 protein species being decreased, including ATP synthase, fatty acid binding-protein, isocitrate dehydrogenase, NADH dehydrogenase, porin, peroxiredoxin, adenylate kinase, tropomyosin, actin, and myosin light chains. Hence, the lack of cardiac dystrophin appears to trigger a generally perturbed protein expression pattern in the MDX heart, affecting especially energy metabolism and contractile proteins.

  9. A defect in dystrophin causes a novel porcine stress syndrome

    Directory of Open Access Journals (Sweden)

    Nonneman Dan J

    2012-06-01

    Full Text Available Abstract Background Losses of slaughter-weight pigs due to transport stress are both welfare and economic concerns to pork producers. Historically, the HAL-1843 mutation in ryanodine receptor 1 was considered responsible for most of the losses; however, DNA testing has effectively eliminated this mutation from commercial herds. We identified two sibling barrows in the USMARC swine herd that died from apparent symptoms of a stress syndrome after transport at 12 weeks of age. The symptoms included open-mouth breathing, skin discoloration, vocalization and loss of mobility. Results We repeated the original mating along with sire-daughter matings to produce additional offspring. At 8 weeks of age, heart rate and electrocardiographs (ECG were monitored during isoflurane anesthesia challenge (3% for 3 min. Four males from the original sire-dam mating and two males from a sire-daughter mating died after one minute of anesthesia. Animals from additional litters were identified as having a stress response, sometimes resulting in death, during regular processing and weighing. Affected animals had elevated plasma creatine phosphokinase (CPK levels before and immediately after isoflurane challenge and cardiac arrhythmias. A pedigree containing 250 pigs, including 49 affected animals, was genotyped with the Illumina PorcineSNP60 Beadchip and only one chromosomal region, SSCX at 25.1-27.7 Mb over the dystrophin gene (DMD, was significantly associated with the syndrome. An arginine to tryptophan (R1958W polymorphism in exon 41 of DMD was the most significant marker associated with stress susceptibility. Immunoblots of affected heart and skeletal muscle showed a dramatic reduction of dystrophin protein and histopathology of affected hearts indicated muscle fiber degeneration. Conclusions A novel stress syndrome was characterized in pigs and the causative genetic factor most likely resides within DMD that results in less dystrophin protein and cardiac

  10. Platelet adhesion: structural and functional diversity of short dystrophin and utrophins in the formation of dystrophin-associated-protein complexes related to actin dynamics.

    Science.gov (United States)

    Cerecedo, Doris; Martínez-Rojas, Dalila; Chávez, Oscar; Martínez-Pérez, Francisco; García-Sierra, Francisco; Rendon, Alvaro; Mornet, Dominique; Mondragón, Ricardo

    2005-12-01

    Platelets are dynamic cell fragments that modify their shape during activation. Utrophin and dystrophins are minor actin-binding proteins present in muscle and non-muscle cytoskeleton. In the present study, we characterised the pattern of Dp71 isoforms and utrophin gene products by immunoblot in human platelets. Two new dystrophin isoforms were found, Dp71f and Dp71 d, as well as the Up71 isoform and the dystrophin-associated proteins, alpha and beta -dystrobrevins. Distribution of Dp71d/Dp71delta110m, Up400/Up71 and dystrophin-associated proteins in relation to the actin cytoskeleton was evaluated by confocal microscopy in both resting and platelets adhered on glass. Formation of two dystrophin-associated protein complexes (Dp71d/Dp71delta110m approximately DAPC and Up400/Up71 approximately DAPC) was demonstrated by co-immunoprecipitation and their distribution in relation to the actin cytoskeleton was characterised during platelet adhesion. The Dp71d/Dp71delta100m approximately DAPC is maintained mainly at the granulomere and is associated with dynamic structures during activation by adhesion to thrombin-coated surfaces. Participation of both Dp71d/Dp71delta110m approximately DAPC and Up400/Up71 approximately DAPC in the biological roles of the platelets is discussed.

  11. Analysis of dystrophin gene deletions by multiplex PCR in eastern India

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    Basak Jayasri

    2006-01-01

    Full Text Available The most common genetic neuromuscular disease of childhood, Duchenne and Becker muscular dystrophy (DMD/BMD is caused by deletion, duplication or point mutation of the dystrophin gene located at Xp 21.2. In the present study DNA from seventy unrelated patients clinically diagnosed as having DMD/BMD referred from different parts of West Bengal, a few other states and Bangladesh are analyzed using the multiplex polymerase chain reaction (m-PCR to screen for exon deletions and its distribution within the dystrophin gene. Out of seventy patients forty six (63% showed large intragenic deletion in the dystrophin gene. About 79% of these deletions are located in the hot spot region i.e., between exon 42 to 53. This is the first report of frequency and distribution of deletion in dystrophin gene in eastern Indian DMD/BMD population.

  12. Dystrophin deficiency exacerbates skeletal muscle pathology in dysferlin-null mice

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    Han Renzhi

    2011-12-01

    Full Text Available Abstract Background Mutations in the genes coding for either dystrophin or dysferlin cause distinct forms of muscular dystrophy. Dystrophin links the cytoskeleton to the sarcolemma through direct interaction with β-dystroglycan. This link extends to the extracellular matrix by β-dystroglycan's interaction with α-dystroglycan, which binds extracellular matrix proteins, including laminin α2, agrin and perlecan, that possess laminin globular domains. The absence of dystrophin disrupts this link, leading to compromised muscle sarcolemmal integrity. Dysferlin, on the other hand, plays an important role in the Ca2+-dependent membrane repair of damaged sarcolemma in skeletal muscle. Because dysferlin and dystrophin play different roles in maintaining muscle cell integrity, we hypothesized that disrupting sarcolemmal integrity with dystrophin deficiency would exacerbate the pathology in dysferlin-null mice and allow further characterization of the role of dysferlin in skeletal muscle. Methods To test our hypothesis, we generated dystrophin/dysferlin double-knockout (DKO mice by breeding mdx mice with dysferlin-null mice and analyzed the effects of a combined deficiency of dysferlin and dystrophin on muscle pathology and sarcolemmal integrity. Results The DKO mice exhibited more severe muscle pathology than either mdx mice or dysferlin-null mice, and, importantly, the onset of the muscle pathology occurred much earlier than it did in dysferlin-deficient mice. The DKO mice showed muscle pathology of various skeletal muscles, including the mandible muscles, as well as a greater number of regenerating muscle fibers, higher serum creatine kinase levels and elevated Evans blue dye uptake into skeletal muscles. Lengthening contractions caused similar force deficits, regardless of dysferlin expression. However, the rate of force recovery within 45 minutes following lengthening contractions was hampered in DKO muscles compared to mdx muscles or dysferlin

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

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    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.

  14. Dystrophin hydrophobic regions in the pathogenesis of Duchenne and Becker muscular dystrophies

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    Yingyin Liang

    2015-05-01

    Full Text Available The aim of our study was to determine the role of dystrophin hydrophobic regions in the pathogenesis of Duchenne (DMD and Becker (BMD muscular dystrophies, by the Kyte-Doolittle scale mean hydrophobicity profile and 3D molecular models. A total of 1038 cases diagnosed with DMD or BMD with the in-frame mutation were collected in our hospital and the Leiden DMD information database in the period 2002-2013. Correlation between clinical types and genotypes were determined on the basis of these two sources. In addition, the Kyte-Doolittle scale mean hydrophobicity of dystrophin was analyzed using BioEdit software and the models of the hydrophobic domains of dystrophin were constructed. The presence of four hydrophobic regions is confirmed. They include the calponin homology CH2 domain on the actin-binding domain (ABD, spectrin-type repeat 16, hinge III and the EF Hand domain. The severe symptoms of DMD usually develop as a result of the mutational disruption in the hydrophobic regions I, II and IV of dystrophin – those that bind associated proteins of the dystrophin-glycoprotein complex (DGC. On the other hand, when the hydrophobic region III is deleted, the connection of the ordered repeat domains of the central rod domain remains intact, resulting in the less severe clinical presentation. We conclude that mutational changes in the structure of hydrophobic regions of dystrophin play an important role in the pathogenesis of DMD.

  15. HEK293 cells express dystrophin Dp71 with nucleus-specific localization of Dp71ab.

    Science.gov (United States)

    Nishida, Atsushi; Yasuno, Sato; Takeuchi, Atsuko; Awano, Hiroyuki; Lee, Tomoko; Niba, Emma Tabe Eko; Fujimoto, Takahiro; Itoh, Kyoko; Takeshima, Yasuhiro; Nishio, Hisahide; Matsuo, Masafumi

    2016-09-01

    The dystrophin gene consists of 79 exons and encodes tissue-specific isoforms. Mutations in the dystrophin gene cause Duchenne muscular dystrophy, of which a substantial proportion of cases are complicated by non-progressive mental retardation. Abnormalities of Dp71, an isoform transcribed from a promoter in intron 62, are a suspected cause of mental retardation. However, the roles of Dp71 in human brain have not been fully elucidated. Here, we characterized dystrophin in human HEK293 cells with the neuronal lineage. Reverse transcription-PCR amplification of the full-length dystrophin transcript revealed the absence of fragments covering the 5' part of the dystrophin cDNA. In contrast, fragments covering exons 64-79 were present. The Dp71 promoter-specific exon G1 was shown spliced to exon 63. We demonstrated that the Dp71 transcript comprised two subisoforms: one lacking exon 78 (Dp71b) and the other lacking both exons 71 and 78 (Dp71ab). Western blotting of cell lysates using an antibody against the dystrophin C-terminal region revealed two bands, corresponding to Dp71b and Dp71ab. Immunohistochemical examination with the dystrophin antibody revealed scattered punctate signals in the cytoplasm and the nucleus. Western blotting revealed one band corresponding to Dp71b in the cytoplasm and two bands corresponding to Dp71b and Dp71ab in the nucleus, with Dp71b being predominant. These results indicated that Dp71ab is a nucleus-specific subisoform. We concluded that Dp71, comprising Dp71b and Dp71ab, was expressed exclusively in HEK293 cells and that Dp71ab was specifically localized to the nucleus. Our findings suggest that Dp71ab in the nucleus contributes to the diverse functions of HEK293 cells.

  16. Loss of dystrophin is associated with increased myocardial stiffness in a model of left ventricular hypertrophy.

    Science.gov (United States)

    Donato, Martín; Buchholz, Bruno; Morales, Celina; Valdez, Laura; Zaobornyj, Tamara; Baratta, Sergio; Paez, Diamela T; Matoso, Mirian; Vaccarino, Guillermo; Chejtman, Demian; Agüero, Oscar; Telayna, Juan; Navia, José; Hita, Alejandro; Boveris, Alberto; Gelpi, Ricardo J

    2017-08-01

    Transition from compensated to decompensated left ventricular hypertrophy (LVH) is accompanied by functional and structural changes. Here, the aim was to evaluate dystrophin expression in murine models and human subjects with LVH by transverse aortic constriction (TAC) and aortic stenosis (AS), respectively. We determined whether doxycycline (Doxy) prevented dystrophin expression and myocardial stiffness in mice. Additionally, ventricular function recovery was evaluated in patients 1 year after surgery. Mice were subjected to TAC and monitored for 3 weeks. A second group received Doxy treatment after TAC. Patients with AS were stratified by normal left ventricular end-diastolic wall stress (LVEDWS) and high LVEDWS, and groups were compared. In mice, LVH decreased inotropism and increased myocardial stiffness associated with a dystrophin breakdown and a decreased mitochondrial O2 uptake (MitoMVO2). These alterations were attenuated by Doxy. Patients with high LVEDWS showed similar results to those observed in mice. A correlation between dystrophin and myocardial stiffness was observed in both mice and humans. Systolic function at 1 year post-surgery was only recovered in the normal-LVEDWS group. In summary, mice and humans present diastolic dysfunction associated with dystrophin degradation. The recovery of ventricular function was observed only in patients with normal LVEDWS and without dystrophin degradation. In mice, Doxy improved MitoMVO2. Based on our results it is concluded that the LVH with high LVEDWS is associated to a degradation of dystrophin and increase of myocardial stiffness. At least in a murine model these alterations were attenuated after the administration of a matrix metalloprotease inhibitor.

  17. Preservation of Muscle Force in Mdx3cv Mice Correlates with Low-Level Expression of a Near Full-Length Dystrophin Protein

    OpenAIRE

    2008-01-01

    The complete absence of dystrophin causes Duchenne muscular dystrophy. Its restoration by greater than 20% is needed to reduce muscle pathology and improve muscle force. Dystrophin levels lower than 20% are considered therapeutically irrelevant but are associated with a less severe phenotype in certain Becker muscular dystrophy patients. To understand the role of low-level dystrophin expression, we compared muscle force and pathology in mdx3cv and mdx4cv mice. Dystrophin was eliminated in mdx...

  18. 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.

  19. Monoclonal antibodies against the muscle-specific N-terminus of dystrophin: Characterization of dystrophin in a muscular dystrophy patient with a frameshift deletion of Exons 3-7

    Energy Technology Data Exchange (ETDEWEB)

    Thanh, L. T.; Man, N. thi; Morris, G.E. (North East Wales Institute, Clwyd (United Kingdom)); Love, D.R.; Davies, K.E. (Institute of Molecular Medicine, John Radcliffe Hospital, Oxford (United Kingdom)); Helliwell, T.R. (Liverpool Univ. (United Kingdom))

    1993-07-01

    The first three exons of the human muscle dystrophin gene were expressed as a [beta]-galactosidase fusion protein. 1-his protein was then used to prepare two monoclonal antibodies (mAbs) which react with native dystrophin on frozen muscle sections and with denatured dystrophin on western blots but which do not cross-react with the distrophin-related protein, utrophin. Both mAbs recognized dystrophin in muscular dystrophy (MD) patients with deletions of exon 3, and further mapping with 11 overlapping synthetic peptides showed that they both recognize an epitope encoded by the muscle-specific exon 1. Neither mAb recognizes the brain dystrophin isoform, confirming the prediction from mRNA data that this has a different N-terminus. One Becker MD patient with a frameshift deletion of exons 3-7 is shown to produce dystrophin which reacts with the N-terminal mAbs, as well as with mAbs which bind on the C-terminal side of the deletion. The data suggest that transcription begins at the normal muscle dystrophin promoter and that the normal reading frame is restored after the deletion. A number of mechanisms have been proposed for restoration of the reading frame after deletion of exons 3-7, but those which predict dystrophin with an abnormal N-terminus do not appear to be major mechanisms in this patient. 27 refs., 6 figs.

  20. 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.

  1. Is the human dystrophin gene's intron structure related to its intron instability?

    Institute of Scientific and Technical Information of China (English)

    盛文利; 陈江瑛; 朱良付; 刘焯霖

    2003-01-01

    Objective To study the human dystrophin gene molecular deletion mechanism, we analyzed breakpoint regions within junction fragments of deletion-type patients and investigated whether the dystrophin gene's intron structure might be related to intron instability.Methods Junction fragments corresponding to exon 46 and 51 deletions were cloned. The breakpoint regions were sequenced, and the features of introns with available Genebank sequences were analyzed.Results An analysis of junction fragment sequences corresponding to exon 46 and 51 deletions showed that all 5' and 3' breakpoints are located within repeat sequences. No small insertions, small deletions, or point mutations are located near the breakpoint junctions. By analyzing the secondary structure of the junction fragments, we demonstrated that all junction fragment breakpoints are located in non-matching regions of single-stranded hairpin loops. A high concentration of repetitive elements is found to be a key feature of many dystrophin introns. In total, 34.8% of the overall dystrophin intron sequences is composed of repeat sequences.Conclusion Repeat elements in many dystrophin gene introns are the key to their structural bases and reflect intron instability. As a result of the primary DNA sequences, single-stranded hairpin loops form, increasing the instability of the gene, and forming the base for breaks in the DNA. The formation of the single-stranded hairpins can result in reattachment of two different breakpoints, producing a deletion.

  2. Dystrophin expression in muscle stem cells regulates their polarity and asymmetric division.

    Science.gov (United States)

    Dumont, Nicolas A; Wang, Yu Xin; von Maltzahn, Julia; Pasut, Alessandra; Bentzinger, C Florian; Brun, Caroline E; Rudnicki, Michael A

    2015-12-01

    Dystrophin is expressed in differentiated myofibers, in which it is required for sarcolemmal integrity, and loss-of-function mutations in the gene that encodes it result in Duchenne muscular dystrophy (DMD), a disease characterized by progressive and severe skeletal muscle degeneration. Here we found that dystrophin is also highly expressed in activated muscle stem cells (also known as satellite cells), in which it associates with the serine-threonine kinase Mark2 (also known as Par1b), an important regulator of cell polarity. In the absence of dystrophin, expression of Mark2 protein is downregulated, resulting in the inability to localize the cell polarity regulator Pard3 to the opposite side of the cell. Consequently, the number of asymmetric divisions is strikingly reduced in dystrophin-deficient satellite cells, which also display a loss of polarity, abnormal division patterns (including centrosome amplification), impaired mitotic spindle orientation and prolonged cell divisions. Altogether, these intrinsic defects strongly reduce the generation of myogenic progenitors that are needed for proper muscle regeneration. Therefore, we conclude that dystrophin has an essential role in the regulation of satellite cell polarity and asymmetric division. Our findings indicate that muscle wasting in DMD not only is caused by myofiber fragility, but also is exacerbated by impaired regeneration owing to intrinsic satellite cell dysfunction.

  3. Parental source effect of inherited mutations in the dystrophin gene of mice and men

    Energy Technology Data Exchange (ETDEWEB)

    Kress, W.; Grimm, T.; Mueller, C.R. [Institute of Human Genetics, Wuerburg (Germany); Bittner, R. [Institute of Anatomy, Wein (Australia)

    1994-09-01

    Skewed X-inactivation has been suspected the genetic cause for some manifesting female carriers of BMD and DMD. To test whether a parental source effect on the protein expression of the dystrophin gene exists, we have set up backcrosses of mdx mice to wild type strains, enabling us to study the effect of the well-defined origin of the mutation on the dystrophin expression. In skeletal muscle sections the immunohistological staining patterns of dystrophin antibodies were showing a significant difference in the proportion of dystrophin positive versus negative fibers, suggesting a lower expression of paternally inherited mdx mutations. These data are in concordance with the pyruvate kinase (PK) levels in the serum: PK levels were much higher when the mutation was of maternal origin as compared to PK levels in paternally derived mutations. In order to test this {open_quotes}paternal source effect{close_quotes} in humans, we checked obligatory carriers of Becker muscular dystrophy (BMD) for the origin of their mutations. Creatin kinase (CK) levels in 21 carriers with maternally derived mutations were compared to CK values from 8 heterozygotes with mutations of paternal origin: CK (mat) = 140.3 IU/1 versus CK (pat) = 48.6 IU/I. The difference is statistically significant at the 5% level. These observations suggest either a differential X-inactivation or an imprinting of the dystrophin gene in mice and men.

  4. 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.

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

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    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.

  6. 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 IK1, 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 IK1 abnormalities in dystrophin-deficient ventricular cardiomyocytes derived from the hearts of Duchenne muscular dystrophy mouse models. We found that IK1 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.

  7. The sarcoglycan-sarcospan complex localization in mouse retina is independent from dystrophins

    Science.gov (United States)

    Fort, Patrice; Estrada, Francisco-Javier; Bordais, Agnès; Mornet, Dominique; Sahel, José-Alain; Picaud, Serge; Vargas, Haydeé Rosas; Coral-Vázquez, Ramón M.; Rendon, Alvaro

    2005-01-01

    The sarcoglycan–sarcospan (SG–SSPN) complex is part of the dystrophin-glycoprotein complex that has been extensively characterized in muscle. To establish the framework for functional studies of sarcoglycans in retina here, we quantified sarcoglycans mRNA levels with real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and performed immunohistochemistry to determine their cellular and subcellular distribution. We showed that the β-, δ-, γ-, ε-sarcoglycans and sarcospan are expressed in mouse retina. They are localized predominantly in the outer and the inner limiting membranes, probably in the Müller cells and also in the ganglion cells axons where the expression of dystrophins have never been reported. We also investigated the status of the sarcoglycans in the retina of mdx3cv mutant mice for all Duchene Muscular Dystrophy (DMD) gene products. The absence of dystrophin did not produce any change in the sarcoglycan–sarcospan components expression and distribution. PMID:15993965

  8. Heteroduplex analysis of the dystrophin gene: application to point mutation and carrier detection.

    Science.gov (United States)

    Prior, T W; Papp, A C; Snyder, P J; Sedra, M S; Western, L M; Bartolo, C; Moxley, R T; Mendell, J R

    1994-03-01

    Approximately one-third of the 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, we 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. We conclude that the technique is not only ideal for mutation detection but is also useful for diagnostic testing.

  9. 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.

  10. Dystrophin expression in a Duchenne muscular dystrophy patient with a frame shift deletion.

    Science.gov (United States)

    Prior, T W; Bartolo, C; Papp, A C; Snyder, P J; Sedra, M S; Burghes, A H; Kissel, J T; Luquette, M H; Tsao, C Y; Mendell, J R

    1997-02-01

    The exon 45 deletion is a common dystrophin gene deletion. Although this is an out-of-frame deletion, which should not allow for protein synthesis, it has been observed in mildly affected patients. We describe a patient with an exon 45 deletion who produced protein, but still had a severe Duchenne muscular dystrophy phenotype. RT-PCR analysis and cDNA sequencing from the muscle biopsy sample revealed that the exon 45 deletion induced exon skipping of exon 44, which resulted in an in-frame deletion and the production of dystrophin. A conformational change in dystrophin induced by the deletion is proposed as being responsible for the severe phenotype in the patient. We feel that the variable clinical phenotype observed in patients with the exon 45 deletion is not due to exon splicing but may be the result of other environmental or genetic factors, or both.

  11. Regional genomic instability predisposes to complex dystrophin gene rearrangements.

    Science.gov (United States)

    Oshima, Junko; Magner, Daniel B; Lee, Jennifer A; Breman, Amy M; Schmitt, Eric S; White, Lisa D; Crowe, Carol A; Merrill, Michelle; Jayakar, Parul; Rajadhyaksha, Aparna; Eng, Christine M; del Gaudio, Daniela

    2009-09-01

    Mutations in the dystrophin gene (DMD) cause Duchenne and Becker muscular dystrophies and the majority of cases are due to DMD gene rearrangements. Despite the high incidence of these aberrations, little is known about their causative molecular mechanism(s). We examined 792 DMD/BMD clinical samples by oligonucleotide array-CGH and report on the junction sequence analysis of 15 unique deletion cases and three complex intragenic rearrangements to elucidate potential underlying mechanism(s). Furthermore, we present three cases with intergenic rearrangements involving DMD and neighboring loci. The cases with intragenic rearrangements include an inversion with flanking deleted sequences; a duplicated segment inserted in direct orientation into a deleted region; and a splicing mutation adjacent to a deletion. Bioinformatic analysis demonstrated that 7 of 12 breakpoints combined among 3 complex cases aligned with repetitive sequences, as compared to 4 of 30 breakpoints for the 15 deletion cases. Moreover, the inversion/deletion case may involve a stem-loop structure that has contributed to the initiation of this rearrangement. For the duplication/deletion and splicing mutation/deletion cases, the presence of the first mutation, either a duplication or point mutation, may have elicited the deletion events in an attempt to correct preexisting mutations. While NHEJ is one potential mechanism for these complex rearrangements, the highly complex junction sequence of the inversion/deletion case suggests the involvement of a replication-based mechanism. Our results support the notion that regional genomic instability, aided by the presence of repetitive elements, a stem-loop structure, and possibly preexisting mutations, may elicit complex rearrangements of the DMD gene.

  12. Exon exchange approach to repair Duchenne dystrophin transcripts.

    Directory of Open Access Journals (Sweden)

    Stéphanie Lorain

    Full Text Available BACKGROUND: Trans-splicing strategies for mRNA repair involve engineered transcripts designed to anneal target mRNAs in order to interfere with their natural splicing, giving rise to mRNA chimeras where endogenous mutated exons have been replaced by exogenous replacement sequences. A number of trans-splicing molecules have already been proposed for replacing either the 5' or the 3' part of transcripts to be repaired. Here, we show the feasibility of RNA surgery by using a double trans-splicing approach allowing the specific substitution of a given mutated exon. METHODOLOGY/PRINCIPAL FINDINGS: As a target we used a minigene encoding a fragment of the mdx dystrophin gene enclosing the mutated exon (exon 23. This minigene was cotransfected with a variety of exon exchange constructions, differing in their annealing domains. We obtained accurate and efficient replacement of exon 23 in the mRNA target. Adding up a downstream intronic splice enhancer DISE in the exon exchange molecule enhanced drastically its efficiency up to 25-45% of repair depending on the construction in use. CONCLUSIONS/SIGNIFICANCE: These results demonstrate the possibility to fix up mutated exons, refurbish deleted exons and introduce protein motifs, while keeping natural untranslated sequences, which are essential for mRNA stability and translation regulation. Conversely to the well-known exon skipping, exon exchange has the advantage to be compatible with almost any type of mutations and more generally to a wide range of genetic conditions. In particular, it allows addressing disorders caused by dominant mutations.

  13. 2′-O-Methyl RNA/Ethylene-Bridged Nucleic Acid Chimera Antisense Oligonucleotides to Induce Dystrophin Exon 45 Skipping

    Directory of Open Access Journals (Sweden)

    Tomoko Lee

    2017-02-01

    Full Text Available Duchenne muscular dystrophy (DMD is a fatal muscle-wasting disease characterized by dystrophin deficiency from mutations in the dystrophin gene. Antisense oligonucleotide (AO-mediated exon skipping targets restoration of the dystrophin reading frame to allow production of an internally deleted dystrophin protein with functional benefit for DMD patients who have out-of-frame deletions. After accelerated US approval of eteplirsen (Exondys 51, which targets dystrophin exon 51 for skipping, efforts are now focused on targeting other exons. For improved clinical benefits, this strategy requires more studies of the delivery method and modification of nucleic acids. We studied a nucleotide with a 2′-O,4′-C-ethylene-bridged nucleic acid (ENA, which shows high nuclease resistance and high affinity for complementary RNA strands. Here, we describe the process of developing a 2′-O-methyl RNA(2′-OMeRNA/ENA chimera AO to induce dystrophin exon 45 skipping. One 18-mer 2′-OMeRNA/ENA chimera (AO85 had the most potent activity for inducing exon 45 skipping in cultured myotubes. AO85 was administered to mdx mice without significant side effects. AO85 transfection into cultured myotubes from 13 DMD patients induced exon 45 skipping in all samples at different levels and dystrophin expression in 11 patients. These results suggest the possible efficacy of AO-mediated exon skipping changes in individual patients and highlight the 2′-OMeRNA/ENA chimera AO as a potential fundamental treatment for DMD.

  14. 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...... of new potential therapeutic targets....

  15. Proteomic Profiling of the Dystrophin-Deficient mdx Phenocopy of Dystrophinopathy-Associated Cardiomyopathy

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    Ashling Holland

    2014-01-01

    Full Text Available Cardiorespiratory complications are frequent symptoms of Duchenne muscular dystrophy, a neuromuscular disorder caused by primary abnormalities in the dystrophin gene. Loss of cardiac dystrophin initially leads to changes in dystrophin-associated glycoproteins and subsequently triggers secondarily sarcolemmal disintegration, fibre necrosis, fibrosis, fatty tissue replacement, and interstitial inflammation. This results in progressive cardiac disease, which is the cause of death in a considerable number of patients afflicted with X-linked muscular dystrophy. In order to better define the molecular pathogenesis of this type of cardiomyopathy, several studies have applied mass spectrometry-based proteomics to determine proteome-wide alterations in dystrophinopathy-associated cardiomyopathy. Proteomic studies included both gel-based and label-free mass spectrometric surveys of dystrophin-deficient heart muscle from the established mdx animal model of dystrophinopathy. Comparative cardiac proteomics revealed novel changes in proteins associated with mitochondrial energy metabolism, glycolysis, signaling, iron binding, antibody response, fibre contraction, basal lamina stabilisation, and cytoskeletal organisation. This review summarizes the importance of studying cardiomyopathy within the field of muscular dystrophy research, outlines key features of the mdx heart and its suitability as a model system for studying cardiac pathogenesis, and discusses the impact of recent proteomic findings for exploring molecular and cellular aspects of cardiac abnormalities in inherited muscular dystrophies.

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

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    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.

  17. Dystrophin and the two related genetic diseases, Duchenne and Becker muscular dystrophies

    Directory of Open Access Journals (Sweden)

    Elisabeth Le Rumeur

    2015-07-01

    Full Text Available Mutations of the dystrophin DMD gene, essentially deletions of one or several exons, are the cause of two devastating and to date incurable diseases, Duchenne (DMD and Becker (BMD muscular dystrophies. Depending upon the preservation or not of the reading frame, dystrophin is completely absent in DMD, or present in either a mutated or a truncated form in BMD. DMD is a severe disease which leads to a premature death of the patients. Therapy approaches are evolving with the aim to transform the severe DMD in the BMD form of the disease by restoring the expression of a mutated or truncated dystrophin. These therapies are based on the assumption that BMD is a mild disease. However, this is not completely true as BMD patients are more or less severely affected and no molecular basis of this heterogeneity of the BMD form of the disease is yet understood. The aim of this review is to report for the correlation between dystrophin structures in BMD deletions in view of this heterogeneity and to emphasize that examining BMD patients in details is highly relevant to anticipate for DMD therapy effects.

  18. 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.

  19. 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

  20. 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

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    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.

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

    National Research Council Canada - National Science Library

    Finkel, Richard S; Flanigan, Kevin M; Wong, Brenda; Bönnemann, Carsten; Sampson, Jacinda; Sweeney, H Lee; Reha, Allen; Northcutt, Valerie J; Elfring, Gary; Barth, Jay; Peltz, Stuart W

    2013-01-01

    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...

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

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    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/.

  3. 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.

  4. A missense mutation in the dystrophin gene in a Duchenne muscular dystrophy patient.

    Science.gov (United States)

    Prior, T W; Papp, A C; Snyder, P J; Burghes, A H; Bartolo, C; Sedra, M S; Western, L M; Mendell, J R

    1993-08-01

    About two thirds of Duchenne muscular dystrophy (DMD) patients have either gene deletions or duplications. The other DMD cases are most likely the result of point mutations that cannot be easily identified by current strategies. Utilizing a heteroduplex technique and direct sequencing of amplified products, we screened our nondeletion/duplication DMD population for point mutations. We now describe what we believe to be the first dystrophin missense mutation in a DMD patient. The mutation results in the substitution of an evolutionarily conserved leucine to arginine in the actin-binding domain. The patient makes a dystrophin protein which is properly localized and is present at a higher level than is observed in DMD patients. This suggests that an intact actin-binding domain is necessary for protein stability and essential for function.

  5. Antisense oligonucleotide induced exon skipping and the dystrophin gene transcript: cocktails and chemistries

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    Fletcher Sue

    2007-07-01

    Full Text Available Abstract Background Antisense oligonucleotides (AOs can interfere with exon recognition and intron removal during pre-mRNA processing, and induce excision of a targeted exon from the mature gene transcript. AOs have been used in vitro and in vivo to redirect dystrophin pre-mRNA processing in human and animal cells. Targeted exon skipping of selected exons in the dystrophin gene transcript can remove nonsense or frame-shifting mutations that would otherwise have lead to Duchenne Muscular Dystrophy, the most common childhood form of muscle wasting. Results Although many dystrophin exons can be excised using a single AO, several exons require two motifs to be masked for efficient or specific exon skipping. Some AOs were inactive when applied individually, yet pronounced exon excision was induced in transfected cells when the AOs were used in select combinations, clearly indicating synergistic rather than cumulative effects on splicing. The necessity for AO cocktails to induce efficient exon removal was observed with 2 different chemistries, 2'-O-methyl modified bases on a phosphorothioate backbone and phosphorodiamidate morpholino oligomers. Similarly, other trends in exon skipping, as a consequence of 2'-O-methyl AO action, such as removal of additional flanking exons or variations in exon skipping efficiency with overlapping AOs, were also seen when the corresponding sequences were prepared as phosphorodiamidate morpholino oligomers. Conclusion The combination of 2 AOs, directed at appropriate motifs in target exons was found to induce very efficient targeted exon skipping during processing of the dystrophin pre-mRNA. This combinatorial effect is clearly synergistic and is not influenced by the chemistry of the AOs used to induce exon excision. A hierarchy in exon skipping efficiency, observed with overlapping AOs composed of 2'-O-methyl modified bases, was also observed when these same sequences were evaluated as phosphorodiamidate morpholino

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

    Science.gov (United States)

    Barnabei, Matthew S; Metzger, Joseph M

    2012-01-01

    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.

  7. 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.

  8. The Dystrophin-Glycoprotein Complex in the Prevention of Muscle Damage

    OpenAIRE

    2011-01-01

    Muscular dystrophies are genetically diverse but share common phenotypic features of muscle weakness, degeneration, and progressive decline in muscle function. Previous work has focused on understanding how disruptions in the dystrophin-glycoprotein complex result in muscular dystrophy, supporting a hypothesis that the muscle sarcolemma is fragile and susceptible to contraction-induced injury in multiple forms of dystrophy. Although benign in healthy muscle, contractions in dystrophic muscle ...

  9. 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.

  10. 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

    , and suggested that significant changes in mdx/ADAM12 muscle might occur post-transcriptionally. Indeed, by immunostaining and immunoblotting we found an approximately 2-fold increase in expression, and distinct extrasynaptic localization, of alpha 7B integrin and utrophin, the functional homolog of dystrophin....... The expression of the dystrophin-associated glycoproteins was also increased. In conclusion, these results demonstrate a novel way to alleviate dystrophin deficiency in mice, and may stimulate the development of new approaches to compensate for dystrophin deficiency in animals and humans....

  11. In silico analyses of dystrophin Dp40 cellular distribution, nuclear export signals and structure modeling

    Directory of Open Access Journals (Sweden)

    Alejandro Martínez-Herrera

    2015-09-01

    Full Text Available Dystrophin Dp40 is the shortest protein encoded by the DMD (Duchenne muscular dystrophy gene. This protein is unique since it lacks the C-terminal end of dystrophins. In this data article, we describe the subcellular localization, nuclear export signals and the three-dimensional structure modeling of putative Dp40 proteins using bioinformatics tools. The Dp40 wild type protein was predicted as a cytoplasmic protein while the Dp40n4 was predicted to be nuclear. Changes L93P and L170P are involved in the nuclear localization of Dp40n4 protein. A close analysis of Dp40 protein scored that amino acids 93LEQEHNNLV101 and 168LLLHDSIQI176 could function as NES sequences and the scores are lost in Dp40n4. In addition, the changes L93/170P modify the tertiary structure of putative Dp40 mutants. The analysis showed that changes of residues 93 and 170 from leucine to proline allow the nuclear localization of Dp40 proteins. The data described here are related to the research article entitled “EF-hand domains are involved in the differential cellular distribution of dystrophin Dp40” (J. Aragón et al. Neurosci. Lett. 600 (2015 115–120 [1].

  12. 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.

  13. 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.

  14. Functional disruption of the dystrophin gene in rhesus monkey using CRISPR/Cas9.

    Science.gov (United States)

    Chen, Yongchang; Zheng, Yinghui; Kang, Yu; Yang, Weili; Niu, Yuyu; Guo, Xiangyu; Tu, Zhuchi; Si, Chenyang; Wang, Hong; Xing, Ruxiao; Pu, Xiuqiong; Yang, Shang-Hsun; Li, Shihua; Ji, Weizhi; Li, Xiao-Jiang

    2015-07-01

    CRISPR/Cas9 has been used to genetically modify genomes in a variety of species, including non-human primates. Unfortunately, this new technology does cause mosaic mutations, and we do not yet know whether such mutations can functionally disrupt the targeted gene or cause the pathology seen in human disease. Addressing these issues is necessary if we are to generate large animal models of human diseases using CRISPR/Cas9. Here we used CRISPR/Cas9 to target the monkey dystrophin gene to create mutations that lead to Duchenne muscular dystrophy (DMD), a recessive X-linked form of muscular dystrophy. Examination of the relative targeting rate revealed that Crispr/Cas9 targeting could lead to mosaic mutations in up to 87% of the dystrophin alleles in monkey muscle. Moreover, CRISPR/Cas9 induced mutations in both male and female monkeys, with the markedly depleted dystrophin and muscle degeneration seen in early DMD. Our findings indicate that CRISPR/Cas9 can efficiently generate monkey models of human diseases, regardless of inheritance patterns. The presence of degenerated muscle cells in newborn Cas9-targeted monkeys suggests that therapeutic interventions at the early disease stage may be effective at alleviating the myopathy.

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

    Science.gov (United States)

    Robinson-Hamm, Jacqueline N; Gersbach, Charles A

    2016-09-01

    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.

  16. 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.

  17. 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.

  18. 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.

  19. A Japanese boy with myalgia and cramps has a novel in-frame deletion of the dystrophin gene.

    Science.gov (United States)

    Ishigaki, C; Patria, S Y; Nishio, H; Yabe, M; Matsuo, M

    1996-05-01

    We report a Japanese Becker muscular dystrophy (BMD) patient with occasional myalgia and cramps during normal activity that developed at the age of 28 months. His family history was negative for neuromuscular diseases. Muscle biopsy analyses, including dystrophin immunostaining, disclosed no clinically relevant findings. The diagnosis of BMD was initially made at the age of 10 years, when indications of persistent high serum levels of CK prompted us to screen deletions in the dystrophin gene by amplification of 19 deletion-prone exons from the genomic DNA by the polymerase chain reaction (PCR). Among the exons examined, exons 13 and 17 were deleted. To clarify the size of the deletion, the dystrophin transcript was analyzed by reverse transcription PCR. The determined nucleotide sequence of the amplified product encompassing exons 10 to 20 disclosed that the entire segment corresponding to exons 13 to 18 (810 bp) was absent, a deletion that would be expected to cause the production of a dystrophin protein lacking 270 amino acids from the rod domain. This result indicates that occasional myalgia and cramps could be early clinical manifestations of mild BMD, especially in patients who have a deletion in the rod domain, and that deletion screening of the dystrophin gene might be the only reliable method to diagnose such cases.

  20. Becker Muscular Dystrophy (BMD) caused by duplication of exons 3-6 of the dystrophin gene presenting as dilated cardiomyopathy

    Energy Technology Data Exchange (ETDEWEB)

    Tsai, A.C.; Allingham-Hawkins, D.J.; Becker, L. [Univ. of Toronto, Ontario (Canada)] [and others

    1994-09-01

    X-linked dilated cardiomyopathy (XLCM) is a progressive myocardial disease presenting with congestive heart failure in teenage males without clinical signs of skeletal myopathy. Tight linkage of XLCM to the DMD locus has been demonstrated; it has been suggested that, at least in some families, XLCM is a {open_quotes}dystrophinopathy.{close_quotes} We report a 14-year-old boy who presented with acute heart failure due to dilated cardiomyopathy. He had no history of muscle weakness, but physical examination revealed pseudohypertrophy of the calf muscles. He subsequently received a heart transplantation. Family history was negative. Serum CK level at the time of diagnosis was 10,416. Myocardial biopsy showed no evidence of carditis. Dystrophin staining of cardiac and skeletal muscle with anti-sera to COOH and NH{sub 2}termini showed a patchy distribution of positivity suggestive of Becker muscular dystrophy. Analysis of 18 of the 79 dystrophin exons detected a duplication that included exons 3-6. The proband`s mother has an elevated serum CK and was confirmed to be a carrier of the same duplication. A mutation in the muscle promotor region of the dystrophin gene has been implicated in the etiology of SLCM. However, Towbin et al. (1991) argued that other 5{prime} mutations in the dystrophin gene could cause selective cardiomyopathy. The findings in our patient support the latter hypothesis. This suggests that there are multiple regions in the dystrophin gene which, when disrupted, can cause isolated dilated cardiomyopathy.

  1. 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.

  2. Analysis of Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    孙淑珍

    2004-01-01

    Chapter Ⅰ Introduction  Sitting in the rocking chair,Carrie dreams her future.This is the deep impression the novel"Sister Carrie"gives us,which is written by Theodore Dreiser(1871-1945),the great American realism writer.  ……

  3. Dystrophin/α1-syntrophin scaffold regulated PLC/PKC-dependent store-operated calcium entry in myotubes.

    Science.gov (United States)

    Sabourin, Jessica; Harisseh, Rania; Harnois, Thomas; Magaud, Christophe; Bourmeyster, Nicolas; Déliot, Nadine; Constantin, Bruno

    2012-12-01

    In skeletal muscles from patient suffering of Duchenne Muscular Dystrophy and from mdx mice, the absence of the cytoskeleton protein dystrophin has been shown to be essential for maintaining a normal calcium influx. We showed that a TRPC store-dependent cation influx is increased by loss of dystrophin or a scaffolding protein α1-syntrophin, however the mechanisms of this calcium mishandling are incompletely understood. First of all, we confirmed that TRPC1 but also STIM1 and Orai1 are supporting the store-operated cation entry which is enhanced in dystrophin-deficient myotubes. Next, we demonstrated that inhibition of PLC or PKC in dystrophin-deficient myotubes restores elevated cation entry to normal levels similarly to enforced minidystrophin expression. In addition, silencing α1-syntrophin also increased cation influx in a PLC/PKC dependent pathway. We also showed that α1-syntrophin and PLCβ are part of a same protein complex reinforcing the idea of their inter-relation in calcium influx regulation. This elevated cation entry was decreased to normal levels by chelating intracellular free calcium with BAPTA-AM. Double treatments with BAPTA-AM and PLC or PKC inhibitors suggested that the elevation of cation influx by PLC/PKC pathway is dependent on cytosolic calcium. All these results demonstrate an involvement in dystrophin-deficient myotubes of a specific calcium/PKC/PLC pathway in elevation of store-operated cation influx supported by the STIM1/Orai1/TRPC1 proteins, which is normally regulated by the α1-syntrophin/dystrophin scaffold.

  4. 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.

  5. 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.

  6. 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

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

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    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.

  9. Current understanding of dystrophin-related muscular dystrophy and therapeutic challenges ahead

    Institute of Scientific and Technical Information of China (English)

    ZHOU Guang-qian; XIE Hui-qi; ZHANG Su-zhen; YANG Zhi-ming

    2006-01-01

    Objective To review the recent research progress in dystrophin-related muscular dystrophy includes X-linked hereditary Duchenne and Becker muscular dystrophies (DMD and BMD).Data sources Information included in this article was identified by searches of PUBMED and other online resources using the key terms DMD, dystrophin, mutations, animal models, pathophysiology, gene expression, stem cells, gene therapy, cell therapy, and pharmacological.Study selection Mainly original milestone articles and timely reviews written by major pioneer investigators of the field were selected.Results The key issues related to the genetic basis and pathophysiological factors of the diseases were critically addressed. The availabilities and advantages of various animal models for the diseases were described. Major molecular and cellular therapeutic approaches were also discussed, many of which have indeed exhibited some success in pre-clinical studies but at the same time encountered a number of technical hurdles, including the efficient and systemic delivery of a functional gene and myogenic precursor/stem cells to repair genetic defects.Conclusions Further understanding of pathophysiological mechanisms at molecular levels and regenerative properites of myogenic precursor/stem cells will promote the development of multiple therapeutic strategies. The combined use of multiple strategies may represent the major challenge as well as the greatest hope for the therapy of these diseases in coming years.

  10. 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.

  11. Membrane Sealant Poloxamer P188 Protects Against Isoproterenol Induced Cardiomyopathy in Dystrophin Deficient Mice

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    Sali Arpana

    2011-05-01

    Full Text Available Abstract Background Cardiomyopathy in Duchenne muscular dystrophy (DMD is an increasing cause of death in patients. The absence of dystrophin leads to loss of membrane integrity, cell death and fibrosis in cardiac muscle. Treatment of cardiomyocyte membrane instability could help prevent cardiomyopathy. Methods Three month old female mdx mice were exposed to the β1 receptor agonist isoproterenol subcutaneously and treated with the non-ionic tri-block copolymer Poloxamer P188 (P188 (460 mg/kg/dose i.p. daily. Cardiac function was assessed using high frequency echocardiography. Tissue was evaluated with Evans Blue Dye (EBD and picrosirius red staining. Results BL10 control mice tolerated 30 mg/kg/day of isoproterenol for 4 weeks while death occurred in mdx mice at 30, 15, 10, 5 and 1 mg/kg/day within 24 hours. Mdx mice tolerated a low dose of 0.5 mg/kg/day. Isoproterenol exposed mdx mice showed significantly increased heart rates (p Conclusions This model suggests that chronic intermittent intraperitoneal P188 treatment can prevent isoproterenol induced cardiomyopathy in dystrophin deficient mdx mice.

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

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    Thibaut Larcher

    Full Text Available 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.

  13. 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.

  14. Sister Carrie in China

    Institute of Scientific and Technical Information of China (English)

    殷希

    2015-01-01

    Sister Carrie has received many Chinese scholar's attention, and it has quantity relevance researches. Therefore, it is valuable to study why it is popular in China and it's education meaning for Chinese people. In addition, to analysis the domestic re-searches and find it's exist problems can help us make a new breakthrough from the study.

  15. Characterization of 65 epitope-specific dystrophin monoclonal antibodies in canine and murine models of duchenne muscular dystrophy by immunostaining and western blot.

    Science.gov (United States)

    Kodippili, Kasun; Vince, Lauren; Shin, Jin-Hong; Yue, Yongping; Morris, Glenn E; McIntosh, Mark A; Duan, Dongsheng

    2014-01-01

    Epitope-specific monoclonal antibodies can provide unique insights for studying cellular proteins. Dystrophin is one of the largest cytoskeleton proteins encoded by 79 exons. The absence of dystrophin results in Duchenne muscular dystrophy (DMD). Over the last two decades, dozens of exon-specific human dystrophin monoclonal antibodies have been developed and successfully used for DMD diagnosis. Unfortunately, the majority of these antibodies have not been thoroughly characterized in dystrophin-deficient dogs, an outstanding large animal model for translational research. To fill the gap, we performed a comprehensive study on 65 dystrophin monoclonal antibodies in normal and dystrophic dogs (heart and skeletal muscle) by immunofluorescence staining and western blot. For comparison, we also included striated muscles from normal BL10 and dystrophin-null mdx mice. Our analysis revealed distinctive species, tissue and assay-dependent recognition patterns of different antibodies. Importantly, we identified 15 antibodies that can consistently detect full-length canine dystrophin in both immunostaining and western blot. Our results will serve as an important reference for studying DMD in the canine model.

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

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    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.

  17. A case of Becker muscular dystrophy resulting from the skipping of four contiguous exons (71-74) of the dystrophin gene during mRNA maturation.

    Science.gov (United States)

    Patria, S Y; Alimsardjono, H; Nishio, H; Takeshima, Y; Nakamura, H; Matsuo, M

    1996-07-01

    The mutations in one-third of both Duchenne and Becker muscular dystrophy patients remain unknown because they do not involve gross rearrangements of the dystrophin gene. Here we report the first example of multiple exon skipping during the splicing of dystrophin mRNA precursor encoded by an apparently normal dystrophin gene. A 9-year-old Japanese boy exhibiting excessive fatigue and high serum creatine kinase activity was examined for dystrophinopathy. An immunohistochemical study of muscle tissue biopsy disclosed faint and discontinuous staining of the N-terminal and rod domains of dystrophin but no staining at all of the C-terminal domain of dystrophin. The dystrophin transcript from muscle tissue was analyzed by the reverse transcriptase polymerase chain reaction. An amplified product encompassing exons 67-79 of dystrophin cDNA was found to be smaller than that of the wild-type product. Sequence analysis of this fragment showed that the 3' end of exon 70 was directly connected to the 5' end of exon 75 and, thus, that exons 71-74 were completely absent. As a result, a truncated dystrophin protein lacking 110 amino acids from the C-terminal domain should result from translation of this truncated mRNA, and the patient was diagnosed as having Becker muscular dystrophy at the molecular level. Genomic DNA was analyzed to identify the cause of the disappearance of these exons. Every exon-encompassing region could be amplified from genomic DNA, indicating that the dystrophin gene is intact. Furthermore, sequencing of these amplified products did not disclose any particular nucleotide change that could be responsible for the multiple exon skipping observed. Considering that exons 71-74 are spliced out alternatively in some tissue-specific isoforms, to suppose that the alternative splicing machinery is present in the muscle tissue of the index case and that it is activated by an undetermined mechanism is reasonable. These results illustrate a novel genetic anomaly that

  18. 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

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

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    Richard S Finkel

    Full Text Available BACKGROUND: 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. METHODS: 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. FINDINGS: 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. INTERPRETATION: 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. TRIAL REGISTRATION: ClinicalTrials.gov NCT00264888.

  20. Bortezomib (PS-341 treatment decreases inflammation and partially rescues the expression of the dystrophin-glycoprotein complex in GRMD dogs.

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    Karla P C Araujo

    Full Text Available Golden retriever muscular dystrophy (GRMD is a genetic myopathy corresponding to Duchenne muscular dystrophy (DMD in humans. Muscle atrophy is known to be associated with degradation of the dystrophin-glycoprotein complex (DGC via the ubiquitin-proteasome pathway. In the present study, we investigated the effect of bortezomib treatment on the muscle fibers of GRMD dogs. Five GRMD dogs were examined; two were treated (TD- Treated dogs with the proteasome inhibitor bortezomib, and three were control dogs (CD. Dogs were treated with bortezomib using the same treatment regimen used for multiple myeloma. Pharmacodynamics were evaluated by measuring the inhibition of 20S proteasome activity in whole blood after treatment and comparing it to that in CD. We performed immunohistochemical studies on muscle biopsy specimens to evaluate the rescue of dystrophin and dystrophin-associated proteins in the muscles of GRMD dogs treated with bortezomib. Skeletal tissue from TD had lower levels of connective tissue deposition and inflammatory cell infiltration than CD as determined by histology, collagen morphometry and ultrastructural analysis. The CD showed higher expression of phospho-NFκB and TGF-β1, suggesting a more pronounced activation of anti-apoptotic factors and inflammatory molecules and greater connective tissue deposition, respectively. Immunohistochemical analysis demonstrated that dystrophin was not present in the sarcoplasmic membrane of either group. However, bortezomib-TD showed higher expression of α- and β-dystroglycan, indicating an improved disease histopathology phenotype. Significant inhibition of 20S proteasome activity was observed 1 hour after bortezomib administration in the last cycle when the dose was higher. Proteasome inhibitors may thus improve the appearance of GRMD muscle fibers, lessen connective tissue deposition and reduce the infiltration of inflammatory cells. In addition, proteasome inhibitors may rescue some

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

    Science.gov (United States)

    Finkel, Richard S; Flanigan, Kevin M; Wong, Brenda; Bönnemann, Carsten; Sampson, Jacinda; Sweeney, H Lee; Reha, Allen; Northcutt, Valerie J; Elfring, Gary; Barth, Jay; Peltz, Stuart W

    2013-01-01

    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.

  2. 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.

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

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    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.

  4. 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

    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....

  5. The Dystrophin-Glycoprotein Complex in the Prevention of Muscle Damage

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    Jessica D. Gumerson

    2011-01-01

    Full Text Available Muscular dystrophies are genetically diverse but share common phenotypic features of muscle weakness, degeneration, and progressive decline in muscle function. Previous work has focused on understanding how disruptions in the dystrophin-glycoprotein complex result in muscular dystrophy, supporting a hypothesis that the muscle sarcolemma is fragile and susceptible to contraction-induced injury in multiple forms of dystrophy. Although benign in healthy muscle, contractions in dystrophic muscle may contribute to a higher degree of muscle damage which eventually overwhelms muscle regeneration capacity. While increased susceptibility of muscle to mechanical injury is thought to be an important contributor to disease pathology, it is becoming clear that not all DGC-associated diseases share this supposed hallmark feature. This paper outlines experimental support for a function of the DGC in preventing muscle damage and examines the evidence that supports novel functions for this complex in muscle that when impaired, may contribute to the pathogenesis of muscular dystrophy.

  6. Proteasome inhibitor (MG132 rescues Nav1.5 protein content and the cardiac sodium current in dystrophin-deficient mdx5cv mice

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    Jean-Sebastien eRougier

    2013-03-01

    Full Text Available The cardiac voltage-gated sodium channel, Nav1.5, plays a central role in cardiac excitability and impulse propagation and associates with the dystrophin multiprotein complex (DMC at the lateral membrane of cardiomyocytes. It was previously shown that Nav1.5 protein content and the sodium current (INa were both decreased in cardiomyocytes of dystrophin-deficient mdx5cv mice. In this study, wild-type (WT and mdx5cv mice were treated for 7 days with the proteasome inhibitor MG132 (10 µg/Kg/24 h using implanted osmotic mini pumps. MG132 rescued both the total amount of Nav1.5 protein and INa but, unlike in previous studies, de novo expression of dystrophin was not observed in skeletal or cardiac muscle. This study suggests that the reduced expression of Nav1.5 in dystrophin-deficient cells is dependent on proteasomal degradation.

  7. Early cardiac failure in a child with Becker muscular dystrophy is due to an abnormally low amount of dystrophin transcript lacking exon 13.

    Science.gov (United States)

    Ishigaki, C; Patria, S Y; Nishio, H; Yoshioka, A; Matsuo, M

    1997-12-01

    Two Japanese brothers with Becker muscular dystrophy were shown by polymerase chain reaction (PCR) and cDNA sequence analysis to produce a dystrophin gene transcript lacking a single exon: that is, number 13. Despite having the same deletion mutation, the brothers showed clearly different clinical phenotypes: the younger brother developed cardiac failure at the age of nine, while the elder brother was asymptomatic. As alternative splicing was not responsible for this clinical difference, the amount of dystrophin transcript was examined by using reverse transcription semi-nested and parallel PCR. The results showed that the amount of the dystrophin transcript in the younger brother was 20% of that of the elder brother. This finding suggested that lesser amount of dystrophin transcript in the younger brother was responsible for the early onset of cardiac failure. This would represent a novel molecular mechanism for dystrophinopathy.

  8. 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.

  9. 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.

  10. 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

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

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    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.

  12. 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 BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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. CONCLUSIONS/SIGNIFICANCE: Together our findings suggest that combined treatment of LANZO and prednisolone attenuates some components of dystrophic pathology in mdx mice. Our findings

  13. "Christian carrying goomies".

    Science.gov (United States)

    1994-01-01

    Dr. Passingan Usurup tells critics of his pragmatic approach on condom promotion that he is a Christian carrying condoms for Christ. He is head of the University of Papua New Guinea Medical Center and is credited with developing an AIDS/HIV policy for the Papua New Guinea Defence Force. The condoms were named Goomy and promoted at launching in 1992 in a blue packet under the slogan "The bond that guards." Goomy was chosen as the name because it is pidgin for rubber, chewing gum, and anything associated with rubber. Blue packets were chosen over the calls of most soldiers for a camouflage design because of its universal appeal as the color of the sea and sky and because it was the preference of women in the airlines. Once firmly ensconced in his role at the University, Usurup plans to develop a policy for students and staff and help to conduct AIDS prevention and education activities on campus. He will encourage students to test for HIV rather than highlighting the gloom and doom of infection and disease.

  14. 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.

  15. Identification of two point mutations and a one base deletion in exon 19 of the dystrophin gene by heteroduplex formation.

    Science.gov (United States)

    Prior, T W; Papp, A C; Snyder, P J; Burghes, A H; Sedra, M S; Western, L M; Bartello, C; Mendell, J R

    1993-03-01

    Two thirds of the Duchenne muscular dystrophy population have either gene deletions or duplications. The nondeletion/duplication cases are most likely the result of point mutations or small deletions and duplications that cannot be easily identified by current strategies. The major obstacle in identifying small mutations is due to the large size of the dystrophin gene. We selectively screened 5 DMD exons containing CpG dinucleotides in 110 DMD patients without detectable deletions or duplications. Nonsenses mutations are frequently due to a C- to -T transition within a CG dinucleotide pair. To screen for the nonsense mutations, we used the heteroduplex method. Utilizing this approach, we identified 2 different nonsense mutations and a single base deletion all occurring in exon 19. This is the first report of a clustering of small mutations in the dystrophin gene.

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

    OpenAIRE

    2015-01-01

    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 exten...

  17. Muscle-specific CRISPR/Cas9 dystrophin gene editing ameliorates pathophysiology in a mouse model for Duchenne muscular dystrophy.

    Science.gov (United States)

    Bengtsson, Niclas E; Hall, John K; Odom, Guy L; Phelps, Michael P; Andrus, Colin R; Hawkins, R David; Hauschka, Stephen D; Chamberlain, Joel R; Chamberlain, Jeffrey S

    2017-02-14

    Gene replacement therapies utilizing adeno-associated viral (AAV) vectors hold great promise for treating Duchenne muscular dystrophy (DMD). A related approach uses AAV vectors to edit specific regions of the DMD gene using CRISPR/Cas9. Here we develop multiple approaches for editing the mutation in dystrophic mdx(4cv) mice using single and dual AAV vector delivery of a muscle-specific Cas9 cassette together with single-guide RNA cassettes and, in one approach, a dystrophin homology region to fully correct the mutation. Muscle-restricted Cas9 expression enables direct editing of the mutation, multi-exon deletion or complete gene correction via homologous recombination in myogenic cells. Treated muscles express dystrophin in up to 70% of the myogenic area and increased force generation following intramuscular delivery. Furthermore, systemic administration of the vectors results in widespread expression of dystrophin in both skeletal and cardiac muscles. Our results demonstrate that AAV-mediated muscle-specific gene editing has significant potential for therapy of neuromuscular disorders.

  18. Localization and quantitation of the chromosome 6-encoded dystrophin-related protein in normal and pathological human muscle.

    Science.gov (United States)

    Karpati, G; Carpenter, S; Morris, G E; Davies, K E; Guerin, C; Holland, P

    1993-03-01

    A dystrophin-related protein (DRP) encoded by a gene on chromosome 6 was studied in 14 normal and 79 pathological human skeletal muscle biopsies, as well as in cultured myotubes by light microscopic immunocytochemistry and quantitative immunoblots. In normal muscle immunoreactive DRP was present at the postjunctional surface membrane, at the surface of satellite cells, in the walls of blood vessels, in Schwann cells and in perineurium of intramuscular nerves. All of this produced a weak signal on immunoblots. In Duchenne/Becker dystrophy (DMD/BMD) and in polymyositis (PM) or dermatomyositis (DM) DRP was present throughout the extrajunctional surface membrane of extra- and intrafusal muscle fibers, particularly regenerating ones. This produced a 15-17-fold increase of DRP over normal in DMD/BMD and 4-10-fold increase over normal in PM and DM on immunoblots. In other pathological muscles, DRP localization pattern and quantity was about the same as in normals. Dystrophin-related protein was present in about the same amounts and distribution in normal and DMD cultured myoblasts and myotubes. The molecular stimulus for the marked upregulation of DRP in DMD/BMD and in the inflammatory myopathies is not known. In DMD/BMD the diffuse sarcolemmal DRP may partially compensate for dystrophin deficiency.

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

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    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.

  20. An alpha-catulin homologue controls neuromuscular function through localization of the dystrophin complex and BK channels in Caenorhabditis elegans.

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    Linu S Abraham

    2010-08-01

    Full Text Available The large conductance, voltage- and calcium-dependent potassium (BK channel serves as a major negative feedback regulator of calcium-mediated physiological processes and has been implicated in muscle dysfunction and neurological disorders. In addition to membrane depolarization, activation of the BK channel requires a rise in cytosolic calcium. Localization of the BK channel near calcium channels is therefore critical for its function. In a genetic screen designed to isolate novel regulators of the Caenorhabditis elegans BK channel, SLO-1, we identified ctn-1, which encodes an α-catulin homologue with homology to the cytoskeletal proteins α-catenin and vinculin. ctn-1 Mutants resemble slo-1 loss-of-function mutants, as well as mutants with a compromised dystrophin complex. We determined that CTN-1 uses two distinct mechanisms to localize SLO-1 in muscles and neurons. In muscles, CTN-1 utilizes the dystrophin complex to localize SLO-1 channels near L-type calcium channels. In neurons, CTN-1 is involved in localizing SLO-1 to a specific domain independent of the dystrophin complex. Our results demonstrate that CTN-1 ensures the localization of SLO-1 within calcium nanodomains, thereby playing a crucial role in muscles and neurons.

  1. 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.

  2. 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

  3. Longitudinal ambulatory measurements of gait abnormality in dystrophin-deficient dogs

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    Voit Thomas

    2011-04-01

    Full Text Available Abstract Background This study aimed to measure the gait abnormalities in GRMD (Golden retriever muscular dystrophy dogs during growth and disease progression using an ambulatory gait analyzer (3D-accelerometers as a possible tool to assess the effects of a therapeutic intervention. Methods Six healthy and twelve GRMD dogs were evaluated twice monthly, from the age of two to nine months. The evolution of each gait variable previously shown to be modified in control and dystrophin-deficient adults was assessed using two-ways variance analysis (age, clinical status with repeated measurements. A principal component analysis (PCA was applied to perfect multivariate data interpretation. Results Speed, stride length, total power and force significantly already decreased (p Conclusion The gait variables measured by the accelerometers were sensitive to early detect and follow the gait disorders and mirrored the heterogeneity of clinical presentations, giving sense to monitor gait in GRMD dogs during progression of the disease and pre-clinical therapeutic trials.

  4. Genetic modifier screens reveal new components that interact with the Drosophila dystroglycan-dystrophin complex.

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    Mariya M Kucherenko

    Full Text Available The Dystroglycan-Dystrophin (Dg-Dys complex has a capacity to transmit information from the extracellular matrix to the cytoskeleton inside the cell. It is proposed that this interaction is under tight regulation; however the signaling/regulatory components of Dg-Dys complex remain elusive. Understanding the regulation of the complex is critical since defects in this complex cause muscular dystrophy in humans. To reveal new regulators of the Dg-Dys complex, we used a model organism Drosophila melanogaster and performed genetic interaction screens to identify modifiers of Dg and Dys mutants in Drosophila wing veins. These mutant screens revealed that the Dg-Dys complex interacts with genes involved in muscle function and components of Notch, TGF-beta and EGFR signaling pathways. In addition, components of pathways that are required for cellular and/or axonal migration through cytoskeletal regulation, such as Semaphorin-Plexin, Frazzled-Netrin and Slit-Robo pathways show interactions with Dys and/or Dg. These data suggest that the Dg-Dys complex and the other pathways regulating extracellular information transfer to the cytoskeletal dynamics are more intercalated than previously thought.

  5. 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.

  6. 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.

  7. 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.

  8. On fast carry select adders

    Science.gov (United States)

    Shamanna, M.; Whitaker, S.

    1992-01-01

    This paper presents an architecture for a high-speed carry select adder with very long bit lengths utilizing a conflict-free bypass scheme. The proposed scheme has almost half the number of transistors and is faster than a conventional carry select adder. A comparative study is also made between the proposed adder and a Manchester carry chain adder which shows that the proposed scheme has the same transistor count, without suffering any performance degradation, compared to the Manchester carry chain adder.

  9. On fast carry select adders

    Science.gov (United States)

    Shamanna, M.; Whitaker, S.

    This paper presents an architecture for a high-speed carry select adder with very long bit lengths utilizing a conflict-free bypass scheme. The proposed scheme has almost half the number of transistors and is faster than a conventional carry select adder. A comparative study is also made between the proposed adder and a Manchester carry chain adder which shows that the proposed scheme has the same transistor count, without suffering any performance degradation, compared to the Manchester carry chain adder.

  10. Nestin expression in end-stage disease in dystrophin-deficient heart: implications for regeneration from endogenous cardiac stem cells.

    Science.gov (United States)

    Berry, Suzanne E; Andruszkiewicz, Peter; Chun, Ju Lan; Hong, Jun

    2013-11-01

    Nestin(+) cardiac stem cells differentiate into striated cells following myocardial infarct. Transplantation of exogenous stem cells into myocardium of a murine model for Duchenne muscular dystrophy (DMD) increased proliferation of endogenous nestin(+) stem cells and resulted in the appearance of nestin(+) striated cells. This correlated with, and may be responsible for, prevention of dilated cardiomyopathy. We examined nestin(+) stem cells in the myocardium of dystrophin/utrophin-deficient (mdx/utrn(-/-)) mice, a model for DMD. We found that 92% of nestin(+) interstitial cells expressed Flk-1, a marker present on cardiac progenitor cells that differentiate into the cardiac lineage, and that a subset expressed Sca-1, present on adult cardiac cells that become cardiomyocytes. Nestin(+) interstitial cells maintained expression of Flk-1 but lost Sca-1 expression with age and were present in lower numbers in dystrophin-deficient heart than in wild-type heart. Unexpectedly, large clusters of nestin(+) striated cells ranging in size from 20 to 250 cells and extending up to 500 μm were present in mdx/utrn(-/-) heart near the end stage of disease. These cells were also present in dystrophin-deficient mdx/utrn(+/-) and mdx heart but not wild-type heart. Nestin(+) striated cells expressed cardiac troponin I, desmin, and Connexin 43 and correlated with proinflammatory CD68(+) macrophages. Elongated nestin(+) interstitial cells with striations were observed that did not express Flk-1 or the late cardiac marker cardiac troponin I but strongly expressed the early cardiac marker desmin. Nestin was also detected in endothelial and smooth muscle cells. These data indicate that new cardiomyocytes form in dystrophic heart, and nestin(+) interstitial cells may generate them in addition to other cells of the cardiac lineage.

  11. Ultrastructural changes in the interstitial cells of Cajal and gastric dysrhythmias in mice lacking full-length dystrophin (mdx mice).

    Science.gov (United States)

    Vannucchi, Maria-Giuliana; Zizzo, Maria-Grazia; Zardo, Claudio; Pieri, Laura; Serio, Rosa; Mulè, Flavia; Faussone-Pellegrini, Maria-Simonetta

    2004-05-01

    At least two populations of c-kit positive interstitial cells of Cajal (ICC) lie in the gastric wall, one located at the myenteric plexus level has a pace-making function and the other located intramuscularly is intermediary in the neurotransmission and regenerates the slow waves. Both of these ICC sub-types express full-length dystrophin. Mdx mice, an animal model lacking in full-length dystrophin and used to study Duchenne muscular dystrophy (DMD), show gastric dismotilities. The aim of the present study was to verify in mdx mice whether: (i) gastric ICC undergo morphological changes, through immunohistochemical and ultrastructural analyses; and (ii) there are alterations in the electrical activity, using intracellular recording technique. In control mice, ICC sub-types showed heterogeneous ultrastructural features, either intramuscularly or at the myenteric plexus level. In mdx mice, all of the ICC sub-types underwent important changes: coated vesicles were significantly more numerous and caveolae significantly fewer than in control; moreover, cytoskeleton and smooth endoplasmic reticulum were reduced and mitochondria enlarged. c-Kit-positivity and integrity of the ICC networks were maintained. In the circular muscle of normal mice slow waves, which consisted of initial and secondary components, occurred with a regular frequency. In mdx mice, slow waves occurred in a highly dysrhythmic fashion and they lacked a secondary component. We conclude that the lack of the full-length dystrophin is associated with ultrastructural modifications of gastric ICC, most of which can be interpreted as signs of new membrane formation and altered Ca(2+) handling, and with defective generation and regeneration of slow wave activity.

  12. 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.

  13. Distribution of dystrophin- and utrophin-associated protein complexes (DAPC/UAPC) in human hematopoietic stem/progenitor cells.

    Science.gov (United States)

    Teniente-De Alba, Carmen; Martínez-Vieyra, Ivette; Vivanco-Calixto, Raúl; Galván, Iván J; Cisneros, Bulmaro; Cerecedo, Doris

    2011-10-01

    Hematopoietic stem cells (HSC) are defined by their cardinal properties, such as sustained proliferation, multilineage differentiation, and self-renewal, which give rise to a hierarchy of progenitor populations with more restricted potential lineage, ultimately leading to the production of all types of mature blood cells. HSC are anchored by cell adhesion molecules to their specific microenvironment, thus regulating their cell cycle, while cell migration is essentially required for seeding the HSC of the fetal bone marrow (BM) during development as well as in adult BM homeostasis. The dystrophin-associated protein complex (DAPC) is a large group of membrane-associated proteins linking the cytoskeleton to the extracellular matrix and exhibiting scaffolding, adhesion, and signaling roles in muscle and non-muscle cells including mature blood cells. Because adhesion and migration are mechanisms that influence the fate of the HSC, we explored the presence and the feasible role of DAPC. In this study, we characterized the pattern expression by immunoblot technique and, by confocal microscopy analysis, the cellular distribution of dystrophin and utrophin gene products, and the dystrophin-associated proteins (α-, β-dystroglycan, α-syntrophin, α-dystrobrevin) in relation to actin filaments in freshly isolated CD34+ cells from umbilical cord blood. Immunoprecipitation assays demonstrated the presence of Dp71d/Dp71Δ110m ∼DAPC and Up400/Up140∼DAPC. The subcellular distribution of the two DAPC in actin-based structures suggests their dynamic participation in adhesion and cell migration. In addition, the particular protein pattern expression found in hematopoietic stem/progenitor cells might be indicative of their feasible participation during differentiation.

  14. Disruption of action potential and calcium signaling properties in malformed myofibers from dystrophin-deficient mice.

    Science.gov (United States)

    Hernández-Ochoa, Erick O; Pratt, Stephen J P; Garcia-Pelagio, Karla P; Schneider, Martin F; Lovering, Richard M

    2015-04-01

    Duchenne muscular dystrophy (DMD), the most common and severe muscular dystrophy, is caused by the absence of dystrophin. Muscle weakness and fragility (i.e., increased susceptibility to damage) are presumably due to structural instability of the myofiber cytoskeleton, but recent studies suggest that the increased presence of malformed/branched myofibers in dystrophic muscle may also play a role. We have previously studied myofiber morphology in healthy wild-type (WT) and dystrophic (MDX) skeletal muscle. Here, we examined myofiber excitability using high-speed confocal microscopy and the voltage-sensitive indicator di-8-butyl-amino-naphthyl-ethylene-pyridinium-propyl-sulfonate (di-8-ANEPPS) to assess the action potential (AP) properties. We also examined AP-induced Ca(2+) transients using high-speed confocal microscopy with rhod-2, and assessed sarcolemma fragility using elastimetry. AP recordings showed an increased width and time to peak in malformed MDX myofibers compared to normal myofibers from both WT and MDX, but no significant change in AP amplitude. Malformed MDX myofibers also exhibited reduced AP-induced Ca(2+) transients, with a further Ca(2+) transient reduction in the branches of malformed MDX myofibers. Mechanical studies indicated an increased sarcolemma deformability and instability in malformed MDX myofibers. The data suggest that malformed myofibers are functionally different from myofibers with normal morphology. The differences seen in AP properties and Ca(2+) signals suggest changes in excitability and remodeling of the global Ca(2+) signal, both of which could underlie reported weakness in dystrophic muscle. The biomechanical changes in the sarcolemma support the notion that malformed myofibers are more susceptible to damage. The high prevalence of malformed myofibers in dystrophic muscle may contribute to the progressive strength loss and fragility seen in dystrophic muscles. © 2015 The Authors. Physiological Reports published by Wiley

  15. Differential expression of myosin heavy chain isoforms in the masticatory muscles of dystrophin-deficient mice.

    Science.gov (United States)

    Spassov, Alexander; Gredes, Tomasz; Gedrange, Tomasz; Lucke, Silke; Morgenstern, Sven; Pavlovic, Dragan; Kunert-Keil, Christiane

    2011-12-01

    The dystrophin-deficient mouse (mdx) is a homologue animal model of Duchenne muscular dystrophy (DMD) and is characterized by slowly progressive muscle weakness accompanied by changes in myosin heavy chain (MyHC) composition. It is likely that the masticatory muscles undergo similar changes. The aim of this study was to examine the masticatory muscles (masseter, temporal, tongue, and soleus) of 100-day-old mdx and control mice (n = 8-10), and the fibre type distribution (by immunohistochemistry) as well as the expression of the corresponding MyHC messenger RNA (mRNA) (protein and mRNA expression, using Western blot or quantitative real-time polymerase chain reaction (RT-PCR)). Immunohistochemistry and western blot analysis revealed that the masticatory muscles in the control and mdx mice consisted mainly of type 2 fibres, whereas soleus muscle consisted of both type 1 and 2 fibres. In the masseter muscle, the mRNA in mdx mice was not different from that found in the controls. However, the mRNA content of the MyHC-2b isoform in mdx mice was lower in comparison with the controls in the temporal muscle [11.9 versus 36.9 per cent; P muscle (65.7 versus 73.8 per cent; P muscle was lower than in the controls (25.9 versus 30.8 per cent; P muscles of mdx mice may lead to changed fibre type composition. The different MyHC gene expression in mdx mice masticatory muscles may be seen as an adaptive mechanism to muscular dystrophy.

  16. Screening the dystrophin gene suggests a high rate of polymorphism in general but no exonic deletions in schizophrenics

    Energy Technology Data Exchange (ETDEWEB)

    Lindor, N.M.; Sobell, J.L.; Thibodeau, S.N. [Mayo Clinic/Foundation, Rochester, MN (United States)] [and others

    1994-03-15

    The dystrophin gene, located at chromosome Xp21, was evaluated as a candidate gene in chronic schizophrenia in response to the report of a large family in which schizophrenia cosegregated with Becker muscular dystrophy. Genomic DNA from 94 men with chronic schizophrenia was evaluated by Southern blot analysis using cDNA probes that span exons 1-59. No exonic deletions were identified. An unexpectedly high rate of polymorphism was calculated in this study and two novel polymorphisms were found, demonstrating the usefulness of the candidate gene approach even when results of the original study are negative. 41 refs., 1 fig., 4 tabs.

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

    Directory of Open Access Journals (Sweden)

    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.

  18. An Analysis of English Carrie

    Institute of Scientific and Technical Information of China (English)

    孙淑珍

    2004-01-01

    @@ Chapter Ⅰ Introduction Sitting in the rocking chair,Carrie dreams her future.This is the deep impression the novel"Sister Carrie"gives us,which is written by Theodore Dreiser(1871-1945),the great American realism writer.

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

    Science.gov (United States)

    Li, Mei; Arner, Anders

    2015-01-01

    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.

  20. 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.

  1. Deletion analysis of the dystrophin gene in Duchenne and Becker muscular dystrophy patients: Use in carrier diagnosis

    Directory of Open Access Journals (Sweden)

    Kumari D

    2003-04-01

    Full Text Available The dystrophin gene was analyzed in 8 Duchenne muscular dystrophy (DMD and 10 Becker muscular dystrophy (BMD unrelated families (22 subjects: 18 index cases and 4 sibs for the presence of deletions by multiplex polymerase chain reaction (mPCR; 27 exons and Southern hybridization using 8 cDMD probes. Deletions were identified in 5 DMD and 7 BMD patients (6 index cases and 1 sib. The concordance between the clinical phenotype and 'reading frame hypothesis' was observed in 11/12 patients (92%. The female relatives of DMD/BMD patients with identifiable deletions were examined by quantitative mPCR. Carriers were identified in 7 families. We also describe a variation in the HindIII pattern with cDNA probe 8 and 11-14. Molecular characterization of the dystrophin gene in this study has been helpful in advising the patients concerning the inheritance of the condition, and carrier diagnosis of female relatives, and should also prove useful for prenatal diagnosis.

  2. 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.

  3. 免疫组织化学dystrophin染色诊断Duchenne型肌营养不良症的研究%Diagnosis of Duchenne muscular dystrophy through dystrophin expression detection by immunohistochemistry

    Institute of Scientific and Technical Information of China (English)

    刘鹏; 沈定国

    2008-01-01

    目的 探讨Duchenne型肌营养不良症(DMD)肌萎缩蛋白(dystrophin)表达规律和临床意义.方法 收集我院7例DMD患者作为试验组,7例非DMD患者为对照组.使用抗dystrophin杆状结构域单抗、免疫组织化学染色,观察肌膜dystrophin表达.结果 7例DMD患者肌细胞膜dystrophin阴性,7例非DMD患者dystrophin染色阳性.结论 证实DMD患者肌细胞膜dystrophin表达阴性,揭示dystrophin缺失是其发病机制,可以作为确诊DMD手段,对临床诊断DMD有实际意义.%Objective To study dystrophin expression in Duchenne muscular dystrophy (DMD) and non-DMD patients. Methods With immunohistochemistry method, using monoclonal antibody of dystrophin, expression of dystrophin was analyzed in 7 DMD patients (experimental group)and 7 non-DMD patients (control group). Results In 7 non-DMD patients, uniform and continuous dystrophin expression was found along the sarolemma, while not in 7 DMD patients. Conclusions Dystrophin expression in myocyte membrane is negative in DMD patients, which indicates that dystrophin loss may be involved in the pathogenesis of DMD. It can be used as a "gold standard" in diagnosing DMD.

  4. 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.

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

    Directory of Open Access Journals (Sweden)

    Sandra Murphy

    2015-09-01

    Full Text Available 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.

  6. 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.

  7. 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.

  8. Absence of Dystrophin Disrupts Skeletal Muscle Signaling: Roles of Ca2+, Reactive Oxygen Species, and Nitric Oxide in the Development of Muscular Dystrophy.

    Science.gov (United States)

    Allen, David G; Whitehead, Nicholas P; Froehner, Stanley C

    2016-01-01

    Dystrophin is a long rod-shaped protein that connects the subsarcolemmal cytoskeleton to a complex of proteins in the surface membrane (dystrophin protein complex, DPC), with further connections via laminin to other extracellular matrix proteins. Initially considered a structural complex that protected the sarcolemma from mechanical damage, the DPC is now known to serve as a scaffold for numerous signaling proteins. Absence or reduced expression of dystrophin or many of the DPC components cause the muscular dystrophies, a group of inherited diseases in which repeated bouts of muscle damage lead to atrophy and fibrosis, and eventually muscle degeneration. The normal function of dystrophin is poorly defined. In its absence a complex series of changes occur with multiple muscle proteins showing reduced or increased expression or being modified in various ways. In this review, we will consider the various proteins whose expression and function is changed in muscular dystrophies, focusing on Ca(2+)-permeable channels, nitric oxide synthase, NADPH oxidase, and caveolins. Excessive Ca(2+) entry, increased membrane permeability, disordered caveolar function, and increased levels of reactive oxygen species are early changes in the disease, and the hypotheses for these phenomena will be critically considered. The aim of the review is to define the early damage pathways in muscular dystrophy which might be appropriate targets for therapy designed to minimize the muscle degeneration and slow the progression of the disease.

  9. Carrying Capacity:An Overview

    Institute of Scientific and Technical Information of China (English)

    Chen Shaofeng

    2004-01-01

    The concept of carrying capacity is derived from ecology, with widespread contentions of its theoretical connotations and applications in the international academic community, especially the impact of human activities on the environment.Disputes on carrying capacity have been occurring not only among biologists and ecologists, but also among mainstream economists. Based on their efforts,the author makes an attempt to describe its origin,connotations, problems, measurement, and at the same time note the latest international progress in this field.

  10. 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.

  11. Distribution of components of basal lamina and dystrophin-dystroglycan complex in the rat pineal gland: differences from the brain tissue and between the subdivisions of the gland.

    Science.gov (United States)

    Bagyura, Zsolt; Pócsai, Károly; Kálmán, Mihály

    2010-01-01

    The pineal gland is an evagination of the brain tissue, a circumventricular neuroendocrine organ. Our immunohistochemical study investigates basal lamina components (laminin, agrin, perlecan, fibronectin), their receptor, the dystrophin-dystroglycan complex (beta-dystroglycan, dystrophin utrophin), aquaporins (-4,-9) and cellular markers (S100, neurofilament, GFAP, glutamine synthetase) in the adult rat corpus pineale. The aim was to compare the immunohistochemical features of the cerebral and pineal vessels and their environment, and to compare their features in the distal and proximal subdivisions of the so-called 'superficial pineal gland'. In contrast to the cerebral vessels, pineal vessels proved to be immunonegative to alpha1-dystrobrevin, but immunoreactive to laminin. An inner, dense, and an outer, loose layer of laminin as two basal laminae were present. The gap between them contained agrin and perlecan. Basal lamina components enmeshed the pinealocytes, too. Components of dystrophin-dystroglycan complex were also distributed along the vessels. Dystrophin, utrophin and agrin gave a 'patchy' distribution rather than a continuous one. The vessels were interconnected by wing-like structures, composed of basal lamina-components: a delicate network forming nests for cells. Cells immunostained with glutamine synthetase, S100-protein or neurofilament protein contacted the vessels, as well as GFAP- or aquaporin-immunostained astrocytes. Within the body a smaller, proximal, GFAP-and aquaporin-containing subdivision, and a larger, distal, GFAP-and aquaporin-free subdivision could be distinguished. The vascular localization of agrin and utrophin, as well as dystrophin, delineated vessels unequally, preferring the proximal or distal end of the body, respectively.

  12. Increased constitutive nitric oxide production by whole body periodic acceleration ameliorates alterations in cardiomyocytes associated with utrophin/dystrophin deficiency.

    Science.gov (United States)

    Lopez, Jose R; Kolster, Juan; Zhang, Rui; Adams, Jose

    2017-07-01

    Duchenne Muscular Dystrophy (DMD) cardiomyopathy is a progressive lethal disease caused by the lack of the dystrophin protein in the heart. The most widely used animal model of DMD is the dystrophin-deficient mdx mouse; however, these mice exhibit a mild dystrophic phenotype with heart failure only late in life. In contrast, mice deficient for both dystrophin and utrophin (mdx/utrn(-/-), or dKO) can be used to model severe DMD cardiomyopathy where pathophysiological indicators of heart failure are detectable by 8-10weeks of age. Nitric oxide (NO) is an important signaling molecule involved in vital functions of regulating rhythm, contractility, and microcirculation of the heart, and constitutive NO production affects the function of proteins involved in excitation-contraction coupling. In this study, we explored the efficacy of enhancing NO production as a therapeutic strategy for treating DMD cardiomyopathy using the dKO mouse model of DMD. Specifically, NO production was induced via whole body periodic acceleration (pGz), a novel non-pharmacologic intervention which enhances NO synthase (NOS) activity through sinusoidal motion of the body in a headward-footward direction, introducing pulsatile shear stress to the vascular endothelium and cardiomyocyte plasma membrane. Male dKO mice were randomized at 8weeks of age to receive daily pGz (480cpm, Gz±3.0m/s(2), 1h/d) for 4weeks or no treatment, and a separate age-matched group of WT animals (pGz-treated and untreated) served as non-diseased controls. At the conclusion of the protocol, cardiomyocytes from untreated dKO animals had, respectively, 4.3-fold and 3.5-fold higher diastolic resting concentration of Ca(2+) ([Ca(2+)]d) and Na(+) ([Na(+)]d) compared to WT, while pGz treatment significantly reduced these levels. For dKO cardiomyocytes, pGz treatment also improved the depressed contractile function, decreased oxidative stress, blunted the elevation in calpain activity, and mitigated the abnormal increase in [Ca

  13. Naturalistic Elements in Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    刘艳晖

    2007-01-01

    @@ Theodore Dreiser is considered to be a controversial writer.His first novel.Sister Carrie makes a new way of presenting re-ality.This paper discusses the naturalistic elements from the de-tailed description of the environment in that society.

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

    Indian Academy of Sciences (India)

    MARYAM HAGHSHENAS; MOHAMMAD TAGHI AKBARI; SHOHREH ZARE KARIZI; FARAVAREH KHORDADPOOR DEILAMANI; SHAHRIAR NAFISSI; ZIVAR SALEHI

    2016-06-01

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

  15. 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.

  16. 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

    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...... 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...... showed dystrophic changes. He had comorbidity with dystonia, slight mental retardation, low stature and neuropathy. The brother of the proband's mother came to medical attention when he was 43 years old. He complained about muscle pain. On examination, a MRC grade 4+ hip extention palsy and a discrete...

  17. 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

    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...... 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...... showed dystrophic changes. He had comorbidity with dystonia, slight mental retardation, low stature and neuropathy. The brother of the proband's mother came to medical attention when he was 43 years old. He complained about muscle pain. On examination, a MRC grade 4+ hip extention palsy and a discrete...

  18. 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.

  19. Ocular and neurodevelopmental features of Duchenne muscular dystrophy: a signature of dystrophin function in the central nervous system.

    Science.gov (United States)

    Ricotti, Valeria; Jägle, Herbert; Theodorou, Maria; Moore, Anthony T; Muntoni, Francesco; Thompson, Dorothy A

    2016-04-01

    Multiple isoforms of dystrophin (Dp427, Dp260, Dp140, Dp71) are expressed differentially in the central nervous system (CNS) including the retinal layers. Disruption of these protein products is responsible for cognitive dysfunction, electroretinogram (ERG) abnormalities and behavioural disorders in Duchenne muscular dystrophy (DMD). We studied the ocular characteristics and neuropsychiatric profile of 16 DMD boys. The ISCEV standard, full-field flash ERGs were assessed. Intellectual ability and behavioural disturbances were measured. All genotypes were associated with mildly abnormal photopic ERG a:b-wave amplitude ratios. In addition, we identified the following genotype/phenotype correlations: boys with mutations upstream of exon 30 (ie, isolated Dp427 altered expression) showed normal scotopic a:b ratios, abnormal photopic oscillatory potential OP2 and normal scotopic OP2. Conversely, all boys with DMD mutations downstream of exon 30 showed profoundly 'negative' scotopic ERGs (a:b ratios >1). In these patients, the involvement of Dp260 isoform resulted in the absence of slow rod pathway signalling in15 Hz scotopic flicker ERGs. These boys had abnormal scotopic OP2 and normal photopic OP2. Finally, children with mutations also affecting Dp71 were associated with more pronounced electronegative ERGs. When correlating ERGs to neurodevelopmental outcome, we found a positive correlation between negative scotopic ERGs and neurodevelopmental disturbances, and the most severe findings were in boys with Dp71 disruption. These findings suggest a strong association between DMD mutations affecting different DMD isoforms with characteristically abnormal scotopic ERGs and severe neurodevelopmental problems. The role of the ERG as a potential biomarker for dystrophin function in the CNS and response to novel genetic therapies warrants further exploration.

  20. 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

    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.

  1. Early manifestation of alteration in cardiac function in dystrophin deficient mdx mouse using 3D CMR tagging

    Directory of Open Access Journals (Sweden)

    Zhong Jia

    2009-10-01

    Full Text Available Abstract Background Duchenne muscular dystrophy (DMD is caused by the absence of the cytoskeletal protein, dystrophin. In DMD patients, dilated cardiomyopathy leading to heart failure may occur during adolescence. However, early cardiac dysfunction is frequently undetected due to physical inactivity and generalized debilitation. The objective of this study is to determine the time course of cardiac functional alterations in mdx mouse, a mouse model of DMD, by evaluating regional ventricular function with CMR tagging. Methods In vivo myocardial function was evaluated by 3D CMR tagging in mdx mice at early (2 months, middle (7 months and late (10 months stages of disease development. Global cardiac function, regional myocardial wall strains, and ventricular torsion were quantified. Myocardial lesions were assessed with Masson's trichrome staining. Results Global contractile indexes were similar between mdx and C57BL/6 mice in each age group. Histology analysis showed that young mdx mice were free of myocardial lesions. Interstitial fibrosis was present in 7 month mdx mice, with further development into patches or transmural lesions at 10 months of age. As a result, 10 month mdx mice showed significantly reduced regional strain and torsion. However, young mdx mice showed an unexpected increase in regional strain and torsion, while 7 month mdx mice displayed similar regional ventricular function as the controls. Conclusion Despite normal global ventricular function, CMR tagging detected a biphasic change in myocardial wall strain and torsion, with an initial increase at young age followed by progressive decrease at older ages. These results suggest that CMR tagging can provide more sensitive measures of functional alterations than global functional indexes in dystrophin-related cardiomyopathies.

  2. 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.

  3. 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.

  4. Properties of Carry Value Transformation

    Directory of Open Access Journals (Sweden)

    Suryakanta Pal

    2012-01-01

    Full Text Available Carry Value Transformation (CVT is a model of discrete deterministic dynamical system. In the present study, it has been proved that (1 the sum of any two nonnegative integers is the same as the sum of their CVT and XOR values. (2 the number of iterations leading to either CVT=0 or XOR=0 does not exceed the maximum of the lengths of the two addenda expressed as binary strings. A similar process of addition of modified Carry Value Transformation (MCVT and XOR requires a maximum of two iterations for MCVT to be zero. (3 an equivalence relation is shown to exist on Z×Z which divides the CV table into disjoint equivalence classes.

  5. 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

    Full Text Available BACKGROUND: 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". METHODOLOGY/PRINCIPAL FINDINGS: 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. CONCLUSIONS/SIGNIFICANCE: 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.

  6. 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

  7. Deficiency of syntrophin, dystroglycan, and merosin in a female infant with a congenital muscular dystrophy phenotype lacking cysteine-rich and C-terminal domains of dystrophin.

    Science.gov (United States)

    Tachi, N; Ohya, K; Chiba, S; Matsuo, M; Patria, S Y; Matsumura, K

    1997-08-01

    Primary deficiency of merosin is the cause of the classic form of congenital muscular dystrophy (CMD) accompanied by brain white matter abnormalities. We report a female infant with dystrophinopathy who was deficient in merosin in skeletal muscle. The patient had a phenotype of typical CMD and white matter abnormalities on brain MRI. Merosin was greatly reduced in the biopsied skeletal muscle. However, the expression of dystroglycan and syntrophin was also greatly reduced, and the immunoreactivity for the antibodies against the cysteine-rich/C-terminal domains of dystrophin was absent in the sarcolemma. Reverse transcriptase polymerase chain reaction analysis of the dystrophin gene revealed a complete lack of exons 71 through 74. In skeletal muscle, only the mutant gene was expressed. These results suggest that the patient is a symptomatic Duchenne muscular dystrophy carrier with skewed X-inactivation. This patient illustrates for the first time that a dystrophin abnormality can cause a secondary deficiency of merosin in dystrophinopathy. The reduction of merosin may account for the clinical phenotype of CMD and correlate with the white matter abnormalities in our patient.

  8. Up-regulation of miR-31 in human atrial fibrillation begets the arrhythmia by depleting dystrophin and neuronal nitric oxide synthase

    Science.gov (United States)

    Carnicer, Ricardo; Recalde, Alice; Muszkiewicz, Anna; Jayaram, Raja; Carena, Maria Cristina; Wijesurendra, Rohan; Stefanini, Matilde; Surdo, Nicoletta C.; Lomas, Oliver; Ratnatunga, Chandana; Sayeed, Rana; Krasopoulos, George; Rajakumar, Timothy; Bueno-Orovio, Alfonso; Verheule, Sander; Fulga, Tudor A.; Rodriguez, Blanca; Schotten, Ulrich

    2016-01-01

    Atrial fibrillation (AF) is a growing public health burden, and its treatment remains a challenge. AF leads to electrical remodeling of the atria, which in turn promotes AF maintenance and resistance to treatment. Although remodeling has long been a therapeutic target in AF, its causes remain poorly understood. We show that atrial-specific up-regulation of microRNA-31 (miR-31) in goat and human AF depletes neuronal nitric oxide synthase (nNOS) by accelerating mRNA decay and alters nNOS subcellular localization by repressing dystrophin translation. By shortening action potential duration and abolishing rate-dependent adaptation of the action potential duration, miR-31 overexpression and/or disruption of nNOS signaling recapitulates features of AF-induced remodeling and significantly increases AF inducibility in mice in vivo. By contrast, silencing miR-31 in atrial myocytes from patients with AF restores dystrophin and nNOS and normalizes action potential duration and its rate dependency. These findings identify atrial-specific up-regulation of miR-31 in human AF as a key mechanism causing atrial dystrophin and nNOS depletion, which in turn contributes to the atrial phenotype begetting this arrhythmia. miR-31 may therefore represent a potential therapeutic target in AF. PMID:27225184

  9. Precise Correction of the Dystrophin Gene in Duchenne Muscular Dystrophy Patient Induced Pluripotent Stem Cells by TALEN and CRISPR-Cas9

    Directory of Open Access Journals (Sweden)

    Hongmei Lisa Li

    2015-01-01

    Full Text Available 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.

  10. Multiplex ligation-dependent probe amplification for rapid detection of deletions and duplications in the dystrophin gene

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective:Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are X-linked disorders caused by mutations in the dystrophin gene. The majority of recognized mutations are copy number changes of individual exons. The objective of the present study was to assess the multiplex ligation-dependent probe amplification (MLPA) effects of detection of gene mutations. Methods: Samples of 20 control males and 80 males and their mothers referred to our diagnostic facility on the clinical suspicion of DMD or BMD were tested by MLPA and multiplex PCR. Results: The mean DQs for all peak of 20 control male samples was 1.02 (range from 0.83 to 1.21) by MLPA. Deletions or duplications were identified in 6 out of 31 families that had been previously tested as negative by multiplex PCR. One case of complex rearrangement involving a duplication of two regions: dupEX3-9 and dupEX 17-41 were found by MLPA. Conclusions: MLPA is a highly sensitive method and rapid alternative to multiplex PCR for detection of DMD and BMD.

  11. Absence of Dystrophin Related Protein-2 disrupts Cajal bands in a patient with Charcot-Marie-Tooth disease.

    Science.gov (United States)

    Brennan, Kathryn M; Bai, Yunhong; Pisciotta, Chiara; Wang, Suola; Feely, Shawna M E; Hoegger, Mark; Gutmann, Laurie; Moore, Steven A; Gonzalez, Michael; Sherman, Diane L; Brophy, Peter J; Züchner, Stephan; Shy, Michael E

    2015-10-01

    Using exome sequencing in an individual with Charcot-Marie-Tooth disease (CMT) we have identified a mutation in the X-linked dystrophin-related protein 2 (DRP2) gene. A 60-year-old gentleman presented to our clinic and underwent clinical, electrophysiological and skin biopsy studies. The patient had clinical features of a length dependent sensorimotor neuropathy with an age of onset of 50 years. Neurophysiology revealed prolonged latencies with intermediate conduction velocities but no conduction block or temporal dispersion. A panel of 23 disease causing genes was sequenced and ultimately was uninformative. Whole exome sequencing revealed a stop mutation in DRP2, c.805C>T (Q269*). DRP2 interacts with periaxin and dystroglycan to form the periaxin-DRP2-dystroglycan complex which plays a role in the maintenance of the well-characterized Cajal bands of myelinating Schwann cells. Skin biopsies from our patient revealed a lack of DRP2 in myelinated dermal nerves by immunofluorescence. Furthermore electron microscopy failed to identify Cajal bands in the patient's dermal myelinated axons in keeping with ultrastructural pathology seen in the Drp2 knockout mouse. Both the electrophysiologic and dermal nerve twig pathology support the interpretation that this patient's DRP2 mutation causes characteristic morphological abnormalities recapitulating the Drp2 knockout model and potentially represents a novel genetic cause of CMT. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. 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.

  13. 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.

  14. Infections That Pets Carry (For Parents)

    Science.gov (United States)

    ... Your 1- to 2-Year-Old Infections That Pets Carry KidsHealth > For Parents > Infections That Pets Carry ... how to protect your family from infections. How Pets Spread Infections Like people, all animals carry germs . ...

  15. A Brief Analysis of Sister Carrie's Character

    Science.gov (United States)

    Yu, Hanying

    2010-01-01

    Carrie is always dreaming while the rocking chair is rocking again and again, this is the deep impression on us after we read "Sister Carrie" which is the first novel of Theodore Dreiser. In this novel the protagonist Sister Carrie is a controversial person. This paper tries to analyze the character of Sister Carrie in order to find out…

  16. Early right ventricular fibrosis and reduction in biventricular cardiac reserve in the dystrophin-deficient mdx heart.

    Science.gov (United States)

    Meyers, Tatyana A; Townsend, DeWayne

    2015-02-15

    Duchenne muscular dystrophy (DMD) is a progressive disease of striated muscle deterioration. Respiratory and cardiac muscle dysfunction are particularly clinically relevant because they result in the leading causes of death in DMD patients. Despite the clinical and physiological significance of these systems, little has been done to understand the cardiorespiratory interaction in DMD. We show here that prior to the onset of global cardiac dysfunction, dystrophin-deficient mdx mice have increased cardiac fibrosis with the right ventricle being particularly affected. Using a novel biventricular cardiac catheterization technique coupled with cardiac stress testing, we demonstrate that both the right and left ventricles have significant reductions in both systolic and diastolic function in response to dobutamine. Unstimulated cardiac function is relatively normal except for a significant reduction in the ventricular pressure transient duration compared with controls. These biventricular analyses also reveal the absence of a dobutamine-induced increase in isovolumic relaxation in the right ventricle of control hearts. Simultaneous assessment of biventricular pressure demonstrates a dobutamine-dependent enhancement of coupling between the ventricles in control mice, which is absent in mdx mice. Furthermore, studies probing the passive-extension properties of the left ventricle demonstrate that the mdx heart is significantly more compliant compared with age-matched C57BL/10 hearts, which have an age-dependent stiffening that is completely absent from dystrophic hearts. These new results indicate that right ventricular fibrosis is an early indicator of the development of dystrophic cardiomyopathy, suggesting a mechanism by which respiratory insufficiency may accelerate the development of heart failure in DMD.

  17. Dystrophin Dp71f associates with the beta1-integrin adhesion complex to modulate PC12 cell adhesion.

    Science.gov (United States)

    Cerna, Joel; Cerecedo, Doris; Ortega, Arturo; García-Sierra, Francisco; Centeno, Federico; Garrido, Efrain; Mornet, Dominique; Cisneros, Bulmaro

    2006-10-01

    Dystrophin Dp71 is the main product of the Duchenne muscular dystrophy gene in the brain; however, its function is unknown. To study the role of Dp71 in neuronal cells, we previously generated by antisense treatment PC12 neuronal cell clones with decreased Dp71 expression (antisense-Dp71 cells). PC12 cells express two different splicing isoforms of Dp71, a cytoplasmic variant called Dp71f and a nuclear isoform called Dp71d. We previously reported that antisense-Dp71 cells display deficient adhesion to substrate and reduced immunostaining of beta1-integrin in the cell area contacting the substrate. In this study, we isolated additional antisense-Dp71 clones to analyze in detail the potential involvement of Dp71f isoform with the beta1-integrin adhesion system of PC12 cells. Immunofluorescence analyses as well as immunoprecipitation assays demonstrated that the PC12 cell beta1-integrin adhesion complex is composed of beta1-integrin, talin, paxillin, alpha-actinin, FAK and actin. In addition, our results showed that Dp71f associates with most of the beta1-integrin complex components (beta1-integrin, FAK, alpha-actinin, talin and actin). In the antisense-Dp71 cells, the deficiency of Dp71 provokes a significant reduction of the beta1-integrin adhesion complex and, consequently, the deficient adhesion of these cells to laminin. In vitro binding experiments confirmed the interaction of Dp71f with FAK and beta1-integrin. Our data indicate that Dp71f is a structural component of the beta1-integrin adhesion complex of PC12 cells that modulates PC12 cell adhesion by conferring proper complex assembly and/or maintenance.

  18. 25 CFR 167.6 - Carrying capacities.

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Carrying capacities. 167.6 Section 167.6 Indians BUREAU... Carrying capacities. (a) The Commissioner of Indian Affairs on June 26, 1943, promulgated the authorized carrying capacity for each land management district of the Navajo Reservation. (b) Recommended...

  19. 7 CFR 1437.402 - Carrying capacity.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 10 2010-01-01 2010-01-01 false Carrying capacity. 1437.402 Section 1437.402... Determining Coverage of Forage Intended for Animal Consumption § 1437.402 Carrying capacity. (a) CCC will establish a carrying capacity for all grazed forage present in the county for purposes of administering...

  20. A FEMINIST READING OF SISTER CARRIE

    Institute of Scientific and Technical Information of China (English)

    高陈科

    2011-01-01

    In the history of American literature, Sister Carrie has always been a controversial character. The critics regard Carrie either as a "fallen woman" or as a "new women". This thesis aims to offer a feminist reading of the image of Sister Carrie in the con

  1. Local Dynamic Stability Associated with Load Carrying

    Directory of Open Access Journals (Sweden)

    Jian Liu

    2013-03-01

    Conclusion: Current study confirmed the sensitivity of local dynamic stability measure in load carrying situation. It was concluded that load carrying tasks were associated with declined local dynamic stability, which may result in increased risk of fall accident. This finding has implications in preventing fall accidents associated with occupational load carrying.

  2. 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.

    2016-01-01

    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. PMID:26435413

  3. 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.

  4. Sister Carrie:A Material Pursuer

    Institute of Scientific and Technical Information of China (English)

    马春花

    2015-01-01

    Sister Carrie dramatized by Dreiser is totally a material pursuer. She is selfish and accumulates money in a crazy way. What she does inevitably centers on materials. Living with Drouet and later Hurstwood, Carrie gets what she wants and enjoys the luxurious life in an easy way. However, with the satisfaction of some of her desires, Carrie ’s desires grow and expand. With enough food and clothes, she needs luxury. Hurstwood’s failure in business leads Carrie to the stage and finally she makes a suc⁃cess and becomes a famous actress in Broadway. She gets more money, but her desires grow even higher. Nothing can satisfy her. In this essay, the author tries to analyze Carrie according to Freud’s and Guo Weilu’s theories and prove that Carrie is totally a material pursuer.

  5. 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 (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.

  6. Measuring Social Carrying Capacity: An Exploratory Study

    OpenAIRE

    López-Bonilla, Jesús Manuel; López-Bonilla, Luis Miguel

    2007-01-01

    The tourist carrying capacity commands a growing interest given that it is closely linked with sustainable tourist development. The justification of the utility of this concept is given by means of a simple and efficient methodological proposal, by analysing the social carrying capacity. To this end, an empirical application is carried out in the Western Andalusia. In some of the cases analysed, the satisfaction of the tourist is found to decline when the levels of the tourist use are higher ...

  7. ENVIRONMENTAL CARRYING CAPACITY BASED ON SPATIAL PLANNING

    OpenAIRE

    Luthfi Muta'ali

    2013-01-01

    The aims of this research were to examine environmental carrying capacity analyzed based on aspects of spatial planning and eco-region. The result showed that Kulonprogo Regency has low value of environmental carrying capacity and can only support as much as 79.81% of its total population. Analysis of variance showed significant difference of environmental carrying capacity of protected and cultivated area. The main factor among 12 variables determining the degree of environmen...

  8. Bivalve carrying capacity in coastal ecosystems

    NARCIS (Netherlands)

    Dame, R.F.; Prins, T.C.

    1998-01-01

    carrying capacity of suspension feeding bivalves in 11 coastal and estuarine ecosystems is examined. Bivalve carrying capacity is defined in terms of water mass residence time, primary production time and bivalve clearance time. Turnover times for the 11 ecosystems are compared both two and three di

  9. Comments on the image of Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    张楠

    2016-01-01

    Thedore Oreiser was one of America's greatest writers and one of his famous masterpieces is Sister Carrie. the heroin of the novel was a country girl who struggled for success and finally became a movie star. Analysis on the image of Carrie is of practical significance to the country girls swarming into the city nowdays in our country.

  10. Carrie Chapman Catt and Woman Suffrage.

    Science.gov (United States)

    Hardesty, Carolyn, Ed.

    1989-01-01

    Most of the material for this issue of the "Goldfinch," which explores the life of Carrie Chapman Catt, came from the archives of the State Historical Society of Iowa. Carrie Chapman Catt (1859-1947) was an Iowan who advocated woman suffrage and spent 26 years actively working for that cause. The issue contains a biography of Catt, and…

  11. 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

  12. Parallelization of Reversible Ripple-carry Adders

    DEFF Research Database (Denmark)

    Thomsen, Michael Kirkedal; Axelsen, Holger Bock

    2009-01-01

    The design of fast arithmetic logic circuits is an important research topic for reversible and quantum computing. A special challenge in this setting is the computation of standard arithmetical functions without the generation of \\emph{garbage}. Here, we present a novel parallelization scheme...... wherein $m$ parallel $k$-bit reversible ripple-carry adders are combined to form a reversible $mk$-bit \\emph{ripple-block carry adder} with logic depth $\\mathcal{O}(m+k)$ for a \\emph{minimal} logic depth $\\mathcal{O}(\\sqrt{mk})$, thus improving on the $mk$-bit ripple-carry adder logic depth $\\mathcal......{O}(m\\cdot k)$. The underlying mechanisms of the parallelization scheme are formally proven correct. We also show designs for garbage-less reversible comparison circuits. We compare the circuit costs of the resulting ripple-block carry adder with known optimized reversible ripple-carry adders in measures...

  13. The Concept of Carrying Capacity in Tourism

    Directory of Open Access Journals (Sweden)

    Josef Zelenka

    2014-05-01

    Full Text Available Carrying capacity is often pragmatically, theoretically as well as purely intuitively considered as a concept in the context of tourism sustainability. The carrying capacity application has the greatest potential in protected areas, in frequently visited cultural and natural attractions, and in relation to sustaining of the lifestyle of the local community and tourism destination potential in general. Despite its importance, partial applications, determination of basic theoretical principles, and specifying connection to the other theoretical concepts in tourism (particularly destination life cycle, LAC concept, visitors management, there still is a rightful opinion of some authors suggesting that there is no consistent theory of tourism carrying capacity. This theory would be the base for sophisticated practical carrying capacity applications. This paper is therefore focused on introduction of the theoretical concept of carrying capacity, which can be discussed and possibly further elaborated.

  14. Sister Carrie, an Adherent of Desires

    Institute of Scientific and Technical Information of China (English)

    裴水妹

    2007-01-01

    Sister Carrie is one of the most controversial characters in American literature.Thought as a "fallen woman" firstly,she was defined as a "new woman" by some critics later. However, by digging into the motivaton behind the whole process of Carrie's "success", the relationship between Carrie and her creator (the author), the social conditions of then American, it can be found that Carrie has never been free-standing on her thought and she has never found her real-sdf even after becoming a famous actress. In a society dominated by mass consumerism Carrie is only an adherent of her own desires. She also is a representative of all those country girls flooded into cities, a symbol and a sacrifice of the urbanization of America in a time countryside was overcome by cities.

  15. Gun Carrying by High School Students in Boston, MA: Does Overestimation of Peer Gun Carrying Matter?

    Science.gov (United States)

    Hemenway, David; Vriniotis, Mary; Johnson, Renee M.; Miller, Matthew; Azrael, Deborah

    2011-01-01

    This paper investigates: (1) whether high school students overestimate gun carrying by their peers, and (2) whether those students who overestimate peer gun carrying are more likely to carry firearms. Data come from a randomly sampled survey conducted in 2008 of over 1700 high school students in Boston, MA. Over 5% of students reported carrying a…

  16. Gun Carrying by High School Students in Boston, MA: Does Overestimation of Peer Gun Carrying Matter?

    Science.gov (United States)

    Hemenway, David; Vriniotis, Mary; Johnson, Renee M.; Miller, Matthew; Azrael, Deborah

    2011-01-01

    This paper investigates: (1) whether high school students overestimate gun carrying by their peers, and (2) whether those students who overestimate peer gun carrying are more likely to carry firearms. Data come from a randomly sampled survey conducted in 2008 of over 1700 high school students in Boston, MA. Over 5% of students reported carrying a…

  17. Carry Select Adder Circuit with A Successively Incremented Carry Number Block

    OpenAIRE

    Deepak; Bal Krishan

    2014-01-01

    This paper reports a conditional carry select (CCS) adder circuit with a successively-incremented-carry-number block (SICNB) structure for low-voltage VLSI implementation. Owing to the successively-incremented-carry-number block (SICNB) structure, the new 16-bit SICNB CCS adder provides a 37% faster speed as compared to the conventional conditional Carry select adder based on the SPICE results

  18. Lack of dystrophin leads to the selective loss of superior cervical ganglion neurons projecting to muscular targets in genetically dystrophic mdx mice.

    Science.gov (United States)

    De Stefano, M Egle; Leone, Lucia; Lombardi, Loredana; Paggi, Paola

    2005-12-01

    Autonomic imbalance is a pathological aspect of Duchenne muscular dystrophy. Here, we show that the sympathetic superior cervical ganglion (SCG) of mdx mice, which lack dystrophin (Dp427), has 36% fewer neurons than that of wild-type animals. Cell loss occurs around P10 and affects those neurons innervating muscular targets (heart and iris), which, differently from the submandibular gland (non-muscular target), are precociously damaged by the lack of Dp427. In addition, although we reveal altered axonal defasciculation in the submandibular gland and reduced terminal sprouting in all SCG target organs, poor adrenergic innervation is observed only in the heart and iris. These alterations, detected as early as P5, when neuronal loss has not yet occurred, suggest that in mdx mice the absence of Dp427 directly impairs the axonal growth and terminal sprouting of sympathetic neurons. However, when these intrinsic alterations combine with structural and/or functional damages of muscular targets, neuronal death occurs.

  19. Effects of carrying methods and box handles on two-person team carrying capacity for females.

    Science.gov (United States)

    Wu, Swei-Pi; Chang, Shu-Yu

    2010-07-01

    This study used a psychophysical approach to examine the effects of carrying methods and the presence or absence of box handles on the maximum acceptable weight carried and resulting responses (heart rate and rating of perceived exertion) in a two-person carrying task. After training, 16 female subjects performed a two-person carrying task at knuckle height for an 8-h work period. Each subject performed 4 different carrying combinations two times. The independent variables were carrying methods (parallel and tandem walking) and box handles (with and without handles). For comparison with two-person carrying, the subjects also performed one-person carrying. The results showed that the maximum acceptable weight carried (MAWC), heart rate (HR), and rating of perceived exertion (RPE) were significantly affected by the presence of box handles. However, the subjects' MAWC, HR, and RPE values were not significantly influenced by the carrying methods. The test-retest reliability of the psychophysical approach was 0.945. The carrying efficiency of two-person carrying was 96.2% of the one-person carrying method. In general, the use of box with handles allows the subjects to carry a higher MAWC (with lower HR and RPE) compared to carrying boxes without handles.

  20. Gravitational waves carrying orbital angular momentum

    CERN Document Server

    Bialynicki-Birula, Iwo

    2015-01-01

    Spinorial formalism is used to map every electromagnetic wave into the gravitational wave (within the linearized gravity). In this way we can obtain the gravitational counterparts of Bessel, Laguerre-Gauss, and other light beams carrying orbital angular momentum.

  1. POPULATION DENSITIES AND THE RANGE-CARRYING ...

    African Journals Online (AJOL)

    CAPACITY FOR LARGE MAMMALS IN QUEEN ELIZABETH ... The highest known densities oflarge terrestrial mammals occur on the grasslands in ...... It is estimated that the proper average biomass carrying capacity for the several herbivore.

  2. Transgenic cassava lines carrying heterologous alternative oxidase ...

    African Journals Online (AJOL)

    To screen positive lines for gene function, leaf lobes from two transgenic lines with a line carrying an empty vector and the wild type were subjected to somatic embryogenesis (SE), a known oxidative ... African Journal of Biotechnology Vol.

  3. Role of mental retardation-associated dystrophin-gene product Dp71 in excitatory synapse organization, synaptic plasticity and behavioral functions.

    Directory of Open Access Journals (Sweden)

    Fatma Daoud

    Full Text Available BACKGROUND: Duchenne muscular dystrophy (DMD is caused by deficient expression of the cytoskeletal protein, dystrophin. One third of DMD patients also have mental retardation (MR, likely due to mutations preventing expression of dystrophin and other brain products of the DMD gene expressed from distinct internal promoters. Loss of Dp71, the major DMD-gene product in brain, is thought to contribute to the severity of MR; however, the specific function of Dp71 is poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: Complementary approaches were used to explore the role of Dp71 in neuronal function and identify mechanisms by which Dp71 loss may impair neuronal and cognitive functions. Besides the normal expression of Dp71 in a subpopulation of astrocytes, we found that a pool of Dp71 colocalizes with synaptic proteins in cultured neurons and is expressed in synaptic subcellular fractions in adult brains. We report that Dp71-associated protein complexes interact with specialized modular scaffolds of proteins that cluster glutamate receptors and organize signaling in postsynaptic densities. We then undertook the first functional examination of the brain and cognitive alterations in the Dp71-null mice. We found that these mice display abnormal synapse organization and maturation in vitro, altered synapse density in the adult brain, enhanced glutamatergic transmission and reduced synaptic plasticity in CA1 hippocampus. Dp71-null mice show selective behavioral disturbances characterized by reduced exploratory and novelty-seeking behavior, mild retention deficits in inhibitory avoidance, and impairments in spatial learning and memory. CONCLUSIONS/SIGNIFICANCE: Results suggest that Dp71 expression in neurons play a regulatory role in glutamatergic synapse organization and function, which provides a new mechanism by which inactivation of Dp71 in association with that of other DMD-gene products may lead to increased severity of MR.

  4. 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

  5. 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.

  6. A Naturalistic Reading of Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    谈月

    2016-01-01

    Sister Carrie is well known as the works in which naturalism attained maturity in America. Up until now, the relevant research on Dreiser and his Sister Carrie abroad and at home is primarily concerned with the frustration of American dream, the naturalistic thoughts and pessimism. The paper attempts to study it from naturalistic point of view and explain how environmental, hereditary factors and the idea of“survival of the fittest”influence Carrie’s fate.

  7. Effects of box handle position and carrying range on bi-manual carrying capacity for females.

    Science.gov (United States)

    Wu, Swei-Pi; Loiu, Yi; Chien, Te Hong

    2015-01-01

    This study utilizes a psychophysical approach to examine the effects on carrying capacity for bi-manual carrying tasks involving different handle positions and carrying ranges. A total of 16 female subjects participated in the experiment in groups of two people, and each group of subjects performed the tasks in a random order with 12 different combinations of carrying task. The independent variables are handle position (upper, middle, lower) and carrying range (F-F: floor height carried to floor height, F-W: floor height carried to waist height, W-W: waist height carried to waist height, W-F: waist height carried to floor height), the dependent variable is the maximum acceptable carried weight (MAWC), heart rate (HR), and the rating of perceived exertion (RPE). The results show that the handle position has a significant effect on MAWC and overall RPE but no significant effect on HR. Carrying range has a significant effect on the MAWC and HR, but no significant effect on overall HR. The handle position and carrying range have a significant interaction on the MAWC and HR. The RPE for different body parts shows significant differences, and the hands feel the most tired. Overall, this study confirms that the lower handle position with the W-W carrying range is the best combination for a two-person carrying task.

  8. Optimal growth trajectories with finite carrying capacity.

    Science.gov (United States)

    Caravelli, F; Sindoni, L; Caccioli, F; Ududec, C

    2016-08-01

    We consider the problem of finding optimal strategies that maximize the average growth rate of multiplicative stochastic processes. For a geometric Brownian motion, the problem is solved through the so-called Kelly criterion, according to which the optimal growth rate is achieved by investing a constant given fraction of resources at any step of the dynamics. We generalize these finding to the case of dynamical equations with finite carrying capacity, which can find applications in biology, mathematical ecology, and finance. We formulate the problem in terms of a stochastic process with multiplicative noise and a nonlinear drift term that is determined by the specific functional form of carrying capacity. We solve the stochastic equation for two classes of carrying capacity functions (power laws and logarithmic), and in both cases we compute the optimal trajectories of the control parameter. We further test the validity of our analytical results using numerical simulations.

  9. Optimal growth trajectories with finite carrying capacity

    Science.gov (United States)

    Caravelli, F.; Sindoni, L.; Caccioli, F.; Ududec, C.

    2016-08-01

    We consider the problem of finding optimal strategies that maximize the average growth rate of multiplicative stochastic processes. For a geometric Brownian motion, the problem is solved through the so-called Kelly criterion, according to which the optimal growth rate is achieved by investing a constant given fraction of resources at any step of the dynamics. We generalize these finding to the case of dynamical equations with finite carrying capacity, which can find applications in biology, mathematical ecology, and finance. We formulate the problem in terms of a stochastic process with multiplicative noise and a nonlinear drift term that is determined by the specific functional form of carrying capacity. We solve the stochastic equation for two classes of carrying capacity functions (power laws and logarithmic), and in both cases we compute the optimal trajectories of the control parameter. We further test the validity of our analytical results using numerical simulations.

  10. Carry Select Adder Circuit with A Successively Incremented Carry Number Block

    Directory of Open Access Journals (Sweden)

    Deepak

    2014-04-01

    Full Text Available This paper reports a conditional carry select (CCS adder circuit with a successively-incremented-carry-number block (SICNB structure for low-voltage VLSI implementation. Owing to the successively-incremented-carry-number block (SICNB structure, the new 16-bit SICNB CCS adder provides a 37% faster speed as compared to the conventional conditional Carry select adder based on the SPICE results

  11. An Optical Carry Chain Fast Adder

    Directory of Open Access Journals (Sweden)

    D. Al-Dabass

    1994-12-01

    Full Text Available A significant problem in Arithmetic Unit design and particularly for systolic arrays remains the speed attainable in achieving high speed addition. The root of the problem is carry propagation and a method is presented which is relatively independent of word length. The problem is addressed by the description of a suggested radical design involving a hybrid optical and electronic approach. The method of carry chain addition through pass gates is explained and a suggested implementation utilising Fabry-Perott resonators, optical waveguides and voltage controlled couplers is described. The design is suitable for n-stage modular expansion.

  12. Error propagation in energetic carrying capacity models

    Science.gov (United States)

    Pearse, Aaron T.; Stafford, Joshua D.

    2014-01-01

    Conservation objectives derived from carrying capacity models have been used to inform management of landscapes for wildlife populations. Energetic carrying capacity models are particularly useful in conservation planning for wildlife; these models use estimates of food abundance and energetic requirements of wildlife to target conservation actions. We provide a general method for incorporating a foraging threshold (i.e., density of food at which foraging becomes unprofitable) when estimating food availability with energetic carrying capacity models. We use a hypothetical example to describe how past methods for adjustment of foraging thresholds biased results of energetic carrying capacity models in certain instances. Adjusting foraging thresholds at the patch level of the species of interest provides results consistent with ecological foraging theory. Presentation of two case studies suggest variation in bias which, in certain instances, created large errors in conservation objectives and may have led to inefficient allocation of limited resources. Our results also illustrate how small errors or biases in application of input parameters, when extrapolated to large spatial extents, propagate errors in conservation planning and can have negative implications for target populations.

  13. Increased carrying capacity with perennial forage kochia

    Science.gov (United States)

    Carrying capacity can be increased on grass-dominated rangeland pastures by including perennial forage kochia (Kochia prostrata) as one of the plant components. The objectives of the study reported here were to compare the differences of traditional winter pastures versus pastures with forage kochi...

  14. Proof-Carrying Code with Correct Compilers

    Science.gov (United States)

    Appel, Andrew W.

    2009-01-01

    In the late 1990s, proof-carrying code was able to produce machine-checkable safety proofs for machine-language programs even though (1) it was impractical to prove correctness properties of source programs and (2) it was impractical to prove correctness of compilers. But now it is practical to prove some correctness properties of source programs, and it is practical to prove correctness of optimizing compilers. We can produce more expressive proof-carrying code, that can guarantee correctness properties for machine code and not just safety. We will construct program logics for source languages, prove them sound w.r.t. the operational semantics of the input language for a proved-correct compiler, and then use these logics as a basis for proving the soundness of static analyses.

  15. Computer Simulation Instruction: Carrying out Chemical Experiments

    Directory of Open Access Journals (Sweden)

    Ibtesam Al-Mashaqbeh

    2014-05-01

    Full Text Available The purpose of this study was to investigate the effect of computer simulation Instruction (CSI on students' achievements: Carrying out chemical experiments to acquire chemical concepts for eleventh grade students. The subject of the study consisted two sections of a one girl's high school in Jordan. One section was randomly assigned to experimental group in which computer simulation Instruction (CSI was used, and the other section was randomly assigned to control group in which students were instructed by using the traditional teaching instruction. The findings indicated that there is progress on the part of the experimental group which used the computer simulation Instruction (CSI and this was reflected positively in the students’ achievement in carrying out chemical experiments to acquire chemical concepts.

  16. A decimal carry-free adder

    Science.gov (United States)

    Nikmehr, Hooman; Phillips, Braden; Lim, Cheng-Chew

    2005-02-01

    Recently, decimal arithmetic has become attractive in the financial and commercial world including banking, tax calculation, currency conversion, insurance and accounting. Although computers are still carrying out decimal calculation using software libraries and binary floating-point numbers, it is likely that in the near future, all processors will be equipped with units performing decimal operations directly on decimal operands. One critical building block for some complex decimal operations is the decimal carry-free adder. This paper discusses the mathematical framework of the addition, introduces a new signed-digit format for representing decimal numbers and presents an efficient architectural implementation. Delay estimation analysis shows that the adder offers improved performance over earlier designs.

  17. Like an eagle carries its young

    Directory of Open Access Journals (Sweden)

    Hans-Georg Wünch

    2016-04-01

    Full Text Available The picture of an eagle carrying its young on its wings (Dt 32:11 is a powerful and encouraging image of trust and security in God. It is particularly relevant for Western culture, where the eagle is a prominent symbol of power and strength. In recent years, though, the translation of the Hebrew term רֶשֶׁנ as ‘eagle’ has come into question and modern exegetes claim that it is more accurately translated as ‘vulture’. But can this really be a symbol of comfort? Furthermore, do eagles (or vultures even carry their young on their wings? This article intends to shed some light on these questions.Keywords: Old Testament; Deuteronomy; Eagle; Vulture

  18. A Feminist Reading of Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    Wang; Jiatong

    2015-01-01

    In the history of American literature,Sister Carrie is the first novel of Theodore Dreiser,it impresses people deeply.Carrie,a poor country girl,becomes a famous star in a big city.She has totally changed from her hard experiences,and she becomes financially independent as a new woman when she goes through hesitation.In he whole novel,the author has planted some strong points of Carrie’s character.At the end of this paper,it analyzes woman’s status in modern time from two aspects of the improvement of female’s social status and the comparison between men and women.

  19. 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

  20. Information carrying capacity of a cosmological constant

    Science.gov (United States)

    Simidzija, Petar; Martín-Martínez, Eduardo

    2017-01-01

    We analyze the exchange of information in different cosmological backgrounds when sender and receiver are timelike separated and communicate through massless fields (without the exchange of light signals). Remarkably, we show that the dominance of a cosmological constant makes the amount of recoverable information imprinted in the field by the sender extremely resilient: it does not decay in time or with the spatial separation of the sender and receiver, and it actually increases with the rate of expansion of the Universe. This is in stark contrast with the information carried by conventional light signals and with previous results on timelike communication through massless fields in matter-dominated cosmologies.

  1. Screening Duchenne and Becker muscular dystrophy patients for deletions in 30 exons of the dystrophin gene by three-multiplex PCR

    Energy Technology Data Exchange (ETDEWEB)

    Risch, N. (Yale Univ., New Haven, CT (United States))

    1992-09-01

    Deletion mutations of the dystrophin gene may cause either the severe Duchenne muscular dystrophy (DMD) or the milder, allelic Becker muscular dystrophy (BMD) and are clustered in two high-frequency-deletion regions (HFDRs) located, respectively, 500 kb and 1,200 kb downstream from the 5[prime] end of the gene. Three PCR reactions described allowed the analysis of a total of 30 exons and led, to the identification of three additional deletions involving the following exons: (a) 42 only, (b) 28-42, and (c) 16 only, none of which were detected with the two original multiplex reactions. Therefore, the three modified multiplexes detected 95 of the 96 deletions identified among the 152 patients studied so far by using Southern analysis and cDNA probes. The only deletion that remained undetected with this system involves exons 22-25 and generates the junction fragment described elsewhere. The percentage of deletion mutations among DMS/BMD patients amounts to 63%, which is in agreement with similar estimates from other laboratories. When field-inversion gel electrophoresis is coupled to Southern analysis, the detection rate of deletion and duplication mutations reaches 65%.

  2. 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.

  3. Activation of Both the Calpain and Ubiquitin-Proteasome Systems Contributes to Septic Cardiomyopathy through Dystrophin Loss/Disruption and mTOR Inhibition

    Science.gov (United States)

    Freitas, Ana Caroline Silva; Figueiredo, Maria Jose; Campos, Erica Carolina; Soave, Danilo Figueiredo; Ramos, Simone Gusmao; Tanowitz, Herbert B.

    2016-01-01

    Cardiac dysfunction caused by the impairment of myocardial contractility has been recognized as an important factor contributing to the high mortality in sepsis. Calpain activation in the heart takes place in response to increased intracellular calcium influx resulting in proteolysis of structural and contractile proteins with subsequent myocardial dysfunction. The purpose of the present study was to test the hypothesis that increased levels of calpain in the septic heart leads to disruption of structural and contractile proteins and that administration of calpain inhibitor-1 (N-acetyl-leucinyl-leucinyl-norleucinal (ALLN)) after sepsis induced by cecal ligation and puncture prevents cardiac protein degradation. We also tested the hypothesis that calpain plays a role in the modulation of protein synthesis/degradation through the activation of proteasome-dependent proteolysis and inhibition of the mTOR pathway. Severe sepsis significantly increased heart calpain-1 levels and promoted ubiquitin and Pa28β over-expression with a reduction in the mTOR levels. In addition, sepsis reduced the expression of structural proteins dystrophin and β-dystroglycan as well as the contractile proteins actin and myosin. ALLN administration prevented sepsis-induced increases in calpain and ubiquitin levels in the heart, which resulted in decreased of structural and contractile proteins degradation and basal mTOR expression levels were re-established. Our results support the concept that increased calpain concentrations may be part of an important mechanism of sepsis-induced cardiac muscle proteolysis. PMID:27880847

  4. Spatio-Temporal Differences in Dystrophin Dynamics at mRNA and Protein Levels Revealed by a Novel FlipTrap Line.

    Directory of Open Access Journals (Sweden)

    Frederique Ruf-Zamojski

    Full Text Available Dystrophin (Dmd is a structural protein that links the extracellular matrix to actin filaments in muscle fibers and is required for the maintenance of muscles integrity. Mutations in Dmd lead to muscular dystrophies in humans and other vertebrates. Here, we report the characterization of a zebrafish gene trap line that fluorescently labels the endogenous Dmd protein (Dmd-citrine, Gt(dmd-citrine ct90a. We show that the Dmd-citrine line recapitulates endogenous dmd transcript expression and Dmd protein localization. Using this Dmd-citrine line, we follow Dmd localization to the myosepta in real-time using time-lapse microscopy, and find that the accumulation of Dmd protein at the transverse myosepta coincides with the onset of myotome formation, a critical stage in muscle maturation. We observed that Dmd protein localizes specifically to the myosepta prior to dmd mRNA localization. Additionally, we demonstrate that the Dmd-citrine line can be used to assess muscular dystrophy following both genetic and physical disruptions of the muscle.

  5. Quantification of the mechanical behavior of carotid arteries from wild-type, dystrophin-deficient, and sarcoglycan-δ knockout mice

    Science.gov (United States)

    Gleason, Rudolph L.; Dye, Wendy W.; Wilson, Emily; Humphrey, Jay D.

    2008-01-01

    As patients with muscular dystrophy live longer because of improved clinical care, they will become increasingly susceptible to many of the cardiovascular diseases that affect the general population. There is, therefore, a pressing need to better understand both the biology and the mechanics of the arterial wall in these patients. In this paper, we use nonlinear constitutive relations to model, for the first time, the biaxial mechanical behavior of carotid arteries from two common mouse models of muscular dystrophy (dystrophin deficient and sarcoglycan-delta null) and wild-type controls. It is shown that a structurally motivated four-fiber family stress-strain relation describes the passive behavior of all three genotypes better than does a commonly used phenomenological exponential model, and that a Rachev-Hayashi model describes the mechanical contribution of smooth muscle contraction under basal tone. Because structurally motivated constitutive relations can be extended easily to model adaptations to altered hemodynamics, results from this study represent an important step toward the ultimate goal of understanding better the mechanobiology and pathophysiology of arteries in muscular dystrophy. PMID:18842267

  6. DESIGN OF OPTIMAL CARRY SKIP ADDER AND CARRY SKIP BCD ADDER USING REVERSIBLE LOGIC GATES

    OpenAIRE

    Praveena Murugesan; Thanushkodi Keppanagounder

    2014-01-01

    Reversible logic circuits have the ability to produce zero power dissipation which has found its importance in quantum computing, optical computing and low power digital circuits. The study presents improved and efficient reversible logic circuits for carry skip adder and carry skip BCD adder. The performance of the proposed architecture is better than the existing works in terms of gate count, garbage outputs and constant inputs. This design forms the basis for different quantum ALU and embe...

  7. Retinal oscillations carry visual information to cortex

    Directory of Open Access Journals (Sweden)

    Kilian Koepsell

    2009-04-01

    Full Text Available Thalamic relay cells fire action potentials that transmit information from retina to cortex. The amount of information that spike trains encode is usually estimated from the precision of spike timing with respect to the stimulus. Sensory input, however, is only one factor that influences neural activity. For example, intrinsic dynamics, such as oscillations of networks of neurons, also modulate firing pattern. Here, we asked if retinal oscillations might help to convey information to neurons downstream. Specifically, we made whole-cell recordings from relay cells to reveal retinal inputs (EPSPs and thalamic outputs (spikes and then analyzed these events with information theory. Our results show that thalamic spike trains operate as two multiplexed channels. One channel, which occupies a low frequency band (<30 Hz, is encoded by average firing rate with respect to the stimulus and carries information about local changes in the visual field over time. The other operates in the gamma frequency band (40-80 Hz and is encoded by spike timing relative to retinal oscillations. At times, the second channel conveyed even more information than the first. Because retinal oscillations involve extensive networks of ganglion cells, it is likely that the second channel transmits information about global features of the visual scene.

  8. Testability Synthesis for Jumping Carry Adders

    Directory of Open Access Journals (Sweden)

    Chien-In Henry Chen

    2002-01-01

    Full Text Available Synthesis for testability ensures that the synthesized circuit is testable by exploring the fundamental relationship between don't care and redundancy. With the exploration of the relationship, redundancy removal can be applied to improve the testability, reduce the area and improve the speed of a synthesized circuit. The test generation problems have been adequately solved, therefore an innovative testability synthesis strategy is necessary for achieving the maximum fault coverage and area reduction for maximum speed. This paper presents a testability synthesis methodology applicable to a top–down design method based on the identification and removal of redundant faults. Emphasis has been placed on the testability synthesis of a high-speed binary jumping carry adder. A synthesized 32-bit testable adder implemented by a 1.2 μm CMOS technology performs addition in 4.09 ns. Comparing with the original synthesized circuit, redundancy removal yields a 100% testable design with a 15% improvement in speed and a 25% reduction in area.

  9. Robust ferromagnetism carried by antiferromagnetic domain walls

    Science.gov (United States)

    Hirose, Hishiro T.; Yamaura, Jun-Ichi; Hiroi, Zenji

    2017-02-01

    Ferroic materials, such as ferromagnetic or ferroelectric materials, have been utilized as recording media for memory devices. A recent trend for downsizing, however, requires an alternative, because ferroic orders tend to become unstable for miniaturization. The domain wall nanoelectronics is a new developing direction for next-generation devices, in which atomic domain walls, rather than conventional, large domains themselves, are the active elements. Here we show that atomically thin magnetic domain walls generated in the antiferromagnetic insulator Cd2Os2O7 carry unusual ferromagnetic moments perpendicular to the wall as well as electron conductivity: the ferromagnetic moments are easily polarized even by a tiny field of 1 mT at high temperature, while, once cooled down, they are surprisingly robust even in an inverse magnetic field of 7 T. Thus, the magnetic domain walls could serve as a new-type of microscopic, switchable and electrically readable magnetic medium which is potentially important for future applications in the domain wall nanoelectronics.

  10. Efficient carry skip Adder design using full adder and carry skip block based on reversible Logic

    Directory of Open Access Journals (Sweden)

    Varun Pratap Singh

    2015-12-01

    Full Text Available In recent years, Reversible Logic is becoming more and more prominent technology having its applications in Quantum Computing, Nanotechnology, and Optical Computing. Reversibility plays an important role when energy efficient computations are considered. In this paper, binary full Adder with Design I and Design II are proposed. The performance analysis is verified using number of reversible gates, Garbage input/outputs, delay, number of logical calculations and Quantum Cost. According to the suitability of full adder design I and design II carry skip adder block is also constructed with some improvement in terms of delay in block carry generation. It is observed that Reversible carry skip Binary Adder with Design II is efficient compared to Design I

  11. 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.

  12. 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.

    Science.gov (United States)

    Martin, Paul T; Golden, Bethannie; Okerblom, Jonathan; Camboni, Marybeth; Chandrasekharan, Kumaran; Xu, Rui; Varki, Ajit; Flanigan, Kevin M; Kornegay, Joe N

    2014-01-01

    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.

  13. Low-Level Laser Therapy (LLLT) in Dystrophin-Deficient Muscle Cells: Effects on Regeneration Capacity, Inflammation Response and Oxidative Stress.

    Science.gov (United States)

    Macedo, Aline Barbosa; Moraes, Luis Henrique Rapucci; Mizobuti, Daniela Sayuri; Fogaça, Aline Reis; Moraes, Fernanda Dos Santos Rapucci; Hermes, Tulio de Almeida; Pertille, Adriana; Minatel, Elaine

    2015-01-01

    The present study evaluated low-level laser therapy (LLLT) effects on some physiological pathways that may lead to muscle damage or regeneration capacity in dystrophin-deficient muscle cells of mdx mice, the experimental model of Duchenne muscular dystrophy (DMD). Primary cultures of mdx skeletal muscle cells were irradiated only one time with laser and analyzed after 24 and 48 hours. The LLLT parameter used was 830 nm wavelengths at 5 J/cm² fluence. The following groups were set up: Ctrl (untreated C57BL/10 primary muscle cells), mdx (untreated mdx primary muscle cells), mdx LA 24 (mdx primary muscle cells - LLLT irradiated and analyzed after 24 h), and mdx LA 48 (mdx primary muscle cells - LLLT irradiated and analyzed after 48 h). The mdx LA 24 and mdx LA 48 groups showed significant increase in cell proliferation, higher diameter in muscle cells and decreased MyoD levels compared to the mdx group. The mdx LA 48 group showed significant increase in Myosin Heavy Chain levels compared to the untreated mdx and mdx LA 24 groups. The mdx LA 24 and mdx LA 48 groups showed significant increase in [Ca2+]i. The mdx group showed significant increase in H2O2 production and 4-HNE levels compared to the Ctrl group and LLLT treatment reduced this increase. GSH levels and GPx, GR and SOD activities increased in the mdx group. Laser treatment reduced the GSH levels and GR and SOD activities in dystrophic muscle cells. The mdx group showed significant increase in the TNF-α and NF-κB levels, which in turn was reduced by the LLLT treatment. Together, these results suggest that the laser treatment improved regenerative capacity and decreased inflammatory response and oxidative stress in dystrophic muscle cells, indicating that LLLT could be a helpful alternative therapy to be associated with other treatment for dystrophinopathies.

  14. Low-Level Laser Therapy (LLLT in Dystrophin-Deficient Muscle Cells: Effects on Regeneration Capacity, Inflammation Response and Oxidative Stress.

    Directory of Open Access Journals (Sweden)

    Aline Barbosa Macedo

    Full Text Available The present study evaluated low-level laser therapy (LLLT effects on some physiological pathways that may lead to muscle damage or regeneration capacity in dystrophin-deficient muscle cells of mdx mice, the experimental model of Duchenne muscular dystrophy (DMD. Primary cultures of mdx skeletal muscle cells were irradiated only one time with laser and analyzed after 24 and 48 hours. The LLLT parameter used was 830 nm wavelengths at 5 J/cm² fluence. The following groups were set up: Ctrl (untreated C57BL/10 primary muscle cells, mdx (untreated mdx primary muscle cells, mdx LA 24 (mdx primary muscle cells - LLLT irradiated and analyzed after 24 h, and mdx LA 48 (mdx primary muscle cells - LLLT irradiated and analyzed after 48 h. The mdx LA 24 and mdx LA 48 groups showed significant increase in cell proliferation, higher diameter in muscle cells and decreased MyoD levels compared to the mdx group. The mdx LA 48 group showed significant increase in Myosin Heavy Chain levels compared to the untreated mdx and mdx LA 24 groups. The mdx LA 24 and mdx LA 48 groups showed significant increase in [Ca2+]i. The mdx group showed significant increase in H2O2 production and 4-HNE levels compared to the Ctrl group and LLLT treatment reduced this increase. GSH levels and GPx, GR and SOD activities increased in the mdx group. Laser treatment reduced the GSH levels and GR and SOD activities in dystrophic muscle cells. The mdx group showed significant increase in the TNF-α and NF-κB levels, which in turn was reduced by the LLLT treatment. Together, these results suggest that the laser treatment improved regenerative capacity and decreased inflammatory response and oxidative stress in dystrophic muscle cells, indicating that LLLT could be a helpful alternative therapy to be associated with other treatment for dystrophinopathies.

  15. Dystrophin Dp71 Isoforms Are Differentially Expressed in the Mouse Brain and Retina: Report of New Alternative Splicing and a Novel Nomenclature for Dp71 Isoforms.

    Science.gov (United States)

    Aragón, Jorge; González-Reyes, Mayram; Romo-Yáñez, José; Vacca, Ophélie; Aguilar-González, Guadalupe; Rendón, Alvaro; Vaillend, Cyrille; Montañez, Cecilia

    2017-01-27

    Multiple dystrophin Dp71 isoforms have been identified in rats, mice, and humans and in several cell line models. These Dp71 isoforms are produced by the alternative splicing of exons 71 to 74 and 78 and intron 77. Three main groups of Dp71 proteins are defined based on their C-terminal specificities: Dp71d, Dp71f, and Dp71e. Dp71 is highly expressed in the brain and retina; however, the specific isoforms present in these tissues have not been determined to date. In this work, we explored the expression of Dp71 isoforms in the mouse brain and retina using RT-PCR assays followed by the cloning of PCR products into the pGEM-T Easy vector, which was used to transform DH5α cells. Dp71-positive colonies were later analyzed by PCR multiplex and DNA sequencing to determine the alternative splicing. We thus demonstrated the expression of Dp71 transcripts corresponding to Dp71, Dp71a, Dp71c, Dp71b, Dp71ab, Dp71 Δ110, and novel Dp71 isoforms spliced in exon 74; 71 and 74; 71, 73 and 74; and 74 and 78, which we named Dp71d Δ74 , Dp71d Δ71,74 , Dp71d Δ71,73-74 , and Dp71f Δ74 , respectively. Additionally, we demonstrated that the Dp71d group of isoforms is highly expressed in the brain, while the Dp71f group predominates in the retina, at both the cDNA and protein levels. These findings suggest that distinct Dp71 isoforms may play different roles in the brain and retina.

  16. Knockdown of dystrophin Dp71 impairs PC12 cells cycle: localization in the spindle and cytokinesis structures implies a role for Dp71 in cell division.

    Directory of Open Access Journals (Sweden)

    Marcela Villarreal-Silva

    Full Text Available The function of dystrophin Dp71 in neuronal cells remains to be established. Previously, we revealed the involvement of this protein in both nerve growth factor (NGF-induced neuronal differentiation and cell adhesion by isolation and characterization of PC12 neuronal cells with depleted levels of Dp71. In this work, a novel phenotype of Dp71-knockdown cells was characterized, which is their delayed growth rate. Cell cycle analyses revealed an altered behavior of Dp71-depleted cells, which consists of a delay in G0/G1 transition and an increase in apoptosis during nocodazole-induced mitotic arrest. Dp71 associates with lamin B1 and β-dystroglycan, proteins involved in aspects of the cell division cycle; therefore, we compared the distribution of Dp71 with that of lamin B1 and β-dystroglycan in PC12 cells at mitosis and cytokinesis by means of immunofluorescence and confocal microscopy analysis. All of these three proteins exhibited a similar immunostaining pattern, localized at mitotic spindle, cleavage furrow, and midbody. It is noteworthy that a drastic decreased staining in mitotic spindle, cleavage furrow, and midbody was observed for both lamin B1 and β-dystroglycan in Dp71-depleted cells. Furthermore, we demonstrated the interaction of Dp71 with lamin B1 in PC12 cells by immunoprecipitation and pull-down assays, and importantly, we revealed that knockdown of Dp71 expression caused a marked reduction in lamin B1 levels and altered localization of the nuclear envelope protein emerin. Our data indicate that Dp71 is a component of the mitotic spindle and cytokinesis multi-protein apparatuses that might modulate the cell division cycle by affecting lamin B1 and β-dystroglycan levels.

  17. microRNA-340-5p Functions Downstream of Cardiotrophin-1 to Regulate Cardiac Eccentric Hypertrophy and Heart Failure via Target Gene Dystrophin.

    Science.gov (United States)

    Zhou, Jian; Gao, Jie; Zhang, Xiaoya; Liu, Yan; Gu, Song; Zhang, Xitao; An, Xiangguang; Yan, Jun; Xin, Yue; Su, Pixiong

    2015-01-01

    Pathological cardiac hypertrophy inevitably leads to the unfavorable outcomes of heart failure (HF) or even sudden death. microRNAs are key regulation factors participating in many pathophysiological processes. Recently, we observed upregulation of microRNA-340-5p (miR-340) in failing human hearts because of dilated cardiomyopathy, but the functional consequence of miR-340 remains to be clarified.We transfected neonatal cardiomyocytes with miR-340 and found fetal gene expression including Nppa, Nppb and Myh7. We also observed eccentric hypertrophy development upon treatment which was analogous to the phenotype after cardiotrophin-1 (CT-1) stimulation. As a potent inducer of cardiac eccentric hypertrophy, treatment by IL-6 family members CT-1 and leukemia inhibitory factor (LIF) led to the elevation of miR-340. Knockdown of miR-340 using antagomir attenuated fetal gene expression and hypertrophy formation, which means miR-340 could convey the hypertrophic signal of CT-1. To demonstrate the initial factor of miR-340 activation, we constructed a volume overloaded abdominal aorta-inferior vena cava fistula rat HF model. miR-340 and CT-1 were found to be up-regulated in the left ventricle. Dystrophin (DMD), a putative target gene of miR-340 which is eccentric hypertrophy-susceptible, was decreased in this HF model upon Western blotting and immunohistochemistry tests. Luciferase assay constructed in two seed sequence of DMD gene 3'UTR showed decreased luciferase activities, and miR-340 transfected cells resulted in the degradation of DMD.miR-340 is a pro-eccentric hypertrophy miRNA, and its expression is dependent on volume overload and cytokine CT-1 activation. Cardiomyocyte structure protein DMD is a target of miR-340.

  18. Efficient Approaches for Designing Fault Tolerant Reversible Carry Look-Ahead and Carry-Skip Adders

    CERN Document Server

    Islam, Md Saiful; begum, Zerina; Hafiz, Mohd Zulfiquar

    2010-01-01

    Combinational or Classical logic circuits dissipate heat for every bit of information that is lost. Information is lost when the input vector cannot be recovered from its corresponding output vector. Reversible logic circuit implements only the functions having one-to-one mapping between its input and output vectors and therefore naturally takes care of heating. Reversible logic design becomes one of the promising research directions in low power dissipating circuit design in the past few years and has found its application in low power CMOS design, digital signal processing and nanotechnology. This paper presents the efficient approaches for designing fault tolerant reversible fast adders that implement carry look-ahead and carry-skip logic. The proposed high speed reversible adders include MIG gates for the realization of its basic building block. The MIG gate is universal and parity preserving. It allows any fault that affects no more than a single signal readily detectable at the circuit's primary outputs...

  19. Fault tolerant reversible logic synthesis: Carry look-ahead and carry-skip adders

    CERN Document Server

    Islam, Md Saiful; Begum, Zerina; Hafiz, Mohd Zulfiquar; 10.1109/ACTEA.2009.5227871

    2010-01-01

    Irreversible logic circuits dissipate heat for every bit of information that is lost. Information is lost when the input vector cannot be recovered from its corresponding output vector. Reversible logic circuit naturally takes care of heating because it implements only the functions that have one-to-one mapping between its input and output vectors. Therefore reversible logic design becomes one of the promising research directions in low power dissipating circuit design in the past few years and has found its application in low power CMOS design, digital signal processing and nanotechnology. This paper presents the efficient approaches for designing reversible fast adders that implement carry look-ahead and carry-skip logic. The proposed 16-bit high speed reversible adder will include IG gates for the realization of its basic building block. The IG gate is universal in the sense that it can be used to synthesize any arbitrary Boolean-functions. The IG gate is parity preserving, that is, the parity of the input...

  20. Carrying Capacity of Marine Region in Liaoning Province

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Carrying capacity is one of important studies on coordinating development of population, resources, and environment. At present, the researches on it mainly concentrate on the carrying capacity for population and economy,such as the water resources carrying capacity, the land resources carrying capacity, the environment carrying capacity,etc. Based on the related theories and methods, this paper creatively proposed the concept and meaning of carrying capacity of marine region, and formed the appraisal system. According to the developing situation of marine economy of Liaoning Province in recent years, and by employing the method of the state space, this paper also measured the cartying capacity and carrying state of marine region and discussed the sustainable problems of marine economy of Liaoning. The research results show that the carrying state of marine region of Liaoning is in the state of overloading at present, but taking a favorable turn.

  1. Delay Efficient 32-Bit Carry-Skip Adder

    OpenAIRE

    Yu Shen Lin; Damu Radhakrishnan

    2008-01-01

    The design of a 32-bit carry-skip adder to achieve minimum delay is presented in this paper. A fast carry look-ahead logic using group generate and group propagate functions is used to speed up the performance of multiple stages of ripple carry adders. The group generate and group propagate functions are generated in parallel with the carry generation for each block. The optimum block sizes are decided by considering the critical path into account. The new architecture ...

  2. Interpretations of the Image of Caroline in Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    钱绘

    2014-01-01

    In the history of American naturalistic literature, Sister Carrie is a representative work of this literary genre. And the protagonist of this novel Caroline has always been a controversial character. Based on Sister Carrie and the social background of that period in America, this paper discussed the various interpretations of the image of Carrie from different aspects.

  3. 46 CFR 111.105-35 - Vessels carrying coal.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Vessels carrying coal. 111.105-35 Section 111.105-35...-GENERAL REQUIREMENTS Hazardous Locations § 111.105-35 Vessels carrying coal. (a) The following are Class II, Division 1, (Zone 10 or Z) locations on a vessel that carries coal: (1) The interior of each...

  4. Delay Efficient 32-Bit Carry-Skip Adder

    Directory of Open Access Journals (Sweden)

    Yu Shen Lin

    2008-01-01

    Full Text Available The design of a 32-bit carry-skip adder to achieve minimum delay is presented in this paper. A fast carry look-ahead logic using group generate and group propagate functions is used to speed up the performance of multiple stages of ripple carry adders. The group generate and group propagate functions are generated in parallel with the carry generation for each block. The optimum block sizes are decided by considering the critical path into account. The new architecture delivers the sum and carry outputs in lesser unit delays than existing carry-skip adders. The adder is implemented in 0.25 m CMOS technology at 3.3 V. The critical delay for the proposed adder is 3.4 nanoseconds. The simulation results show that the proposed adder is 18% faster than the current fastest carry-skip adder.

  5. Regional carrying capacity: case studies of Bohai Rim area

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Carrying capacity research has been carried out for a long time. However, synthesized carrying capacity researches based on systematic views began only in the 1970s. There is even less work done in China. This paper tries to address both synthesized carrying capacity research and its utilization in China. State spaces method from the systematic science was borrowed to construct the conceptual model of regional carrying capacity. Based on the conceptual model and the surveys in the Bohai Rim area, we construct a representative indicators system for quantifying regional carrying capacity in the Bohai Rim. While employing system dynamic models we simulated the evolving trend of both the regional carrying states and regional carrying capacity from 1999 to 2015. The results proved the statement that Bohai Rim is overall over-capacity for a long time and will be over-capacity in the foreseeable future. Among all the restriction factors, water shortage and environmental pollution stand out to be the two primary obstacles for Bohai Rim's sustainable development.Regional differentiation analysis further indicates that coastal areas of the Bohai Rim burden more than its overall level. However, Shandong province shows some good signs in addressing the regional carrying capacity issues. The research is successful in addressing the quantification of regional carrying capacity issues, but nonetheless it needs further refinery and more information.

  6. High performance pipelined multiplier with fast carry-save adder

    Science.gov (United States)

    Wu, Angus

    1990-01-01

    A high-performance pipelined multiplier is described. Its high performance results from the fast carry-save adder basic cell which has a simple structure and is suitable for the Gate Forest semi-custom environment. The carry-save adder computes the sum and carry within two gate delay. Results show that the proposed adder can operate at 200 MHz for a 2-micron CMOS process; better performance is expected in a Gate Forest realization.

  7. Trusted Module Acquisition Through Proof-Carrying Hardware Intellectual Property

    Science.gov (United States)

    2015-05-22

    hardware intellectual property (PCHIP) framework, which aims to ensure the trustworthiness of third-party hardware IPs utilizing formal methods. We...published in non peer-reviewed journals: Final Report: Trusted Module Acquisition Through Proof-Carrying Hardware Intellectual Property Report Title By...borrowing ideas from the proof carrying code (PCC) in software domain, in this project we introduced the proof carrying hardware intellectual property

  8. Optimal design method for fast carry-skip adders

    Science.gov (United States)

    Lee, Songjun; Swartzlander, Earl E., Jr.

    2001-11-01

    A carry-skip adder is faster than a ripple carry adder and it has a simple structure. To maximize the speed it is necessary to optimize the width of the blocks that comprise the carry skip adder. This paper presents a simple algorithm to select the size of each block. Assuming that each logic gate has a unit delay, the algorithm achieves slightly faster designs for 64 and 128 bit adders than previous methods developed by Guyot, et al. and Kantabutra.

  9. Organization and carrying out the triathlon competitions in Ukraine

    Directory of Open Access Journals (Sweden)

    Volodymyr Vodlozerov

    2016-02-01

    Full Text Available Purpose: the aim is analyzing of system of organization and carrying out the triathlon competitions in Ukraine in accordance with rules of triathlon international federation. Material & Methods: comparative analysis of process of organization and carrying out the triathlon competitions in the world and Ukraine was carried out on basis of specialist literature studying, normative base of sports organizations (triathlon federation. Results: inconsistencies were identified in competitions carried out in cold season, particularity of triathlon that intends overcoming the combined distance without time durations between stages. Conclusions: recommendation in eliminate inconsistencies that affect to performance of triathlon competitions in Ukraine was suggested.

  10. Magnetically Guiding Atoms with Current-Carrying Conductors

    Institute of Scientific and Technical Information of China (English)

    刘南春; 高伟建; 印建平

    2002-01-01

    We propose a novel magnetic guide for cold neutral atoms using some current-carrying conductors. The spatial distributions of the magnetic fields from a V-shaped or U-shaped current-carrying conductor are calculated, and the relationship between the resulting magnetic field and the parameters of the current-carrying conductors is analysed in detail. The result shows that these current-carrying conductors can be used to realize a single or a controllable double magnetic guide of cold atoms in the weak-field-seeking state, and to construct various atom-optical elements, and even to realize a single-mode atomic waveguiding under certain conditions.

  11. Training to Increase Safe Tray Carrying among Cocktail Servers

    Science.gov (United States)

    Scherrer, Megan D.; Wilder, David A.

    2008-01-01

    We evaluated the effects of training on proper carrying techniques among 3 cocktail servers to increase safe tray carrying on the job and reduce participants' risk of developing musculoskeletal disorders. As participants delivered drinks to their tables, their finger, arm, and neck positions were observed and recorded. Each participant received…

  12. 46 CFR 185.340 - Vessels carrying vehicles.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Vessels carrying vehicles. 185.340 Section 185.340... TONS) OPERATIONS Miscellaneous Operating Requirements § 185.340 Vessels carrying vehicles. (a) Automobiles or other vehicles must be stowed in such a manner as to permit both passengers and crew to get...

  13. 46 CFR 122.340 - Vessels carrying vehicles.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Vessels carrying vehicles. 122.340 Section 122.340... Miscellaneous Operating Requirements § 122.340 Vessels carrying vehicles. (a) Automobiles or other vehicles must... vehicles freely in the event of fire or other disaster. The decks, where necessary, must be...

  14. 25 CFR 11.444 - Carrying concealed weapons.

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Carrying concealed weapons. 11.444 Section 11.444 Indians... ORDER CODE Criminal Offenses § 11.444 Carrying concealed weapons. A person who goes about in public places armed with a dangerous weapon concealed upon his or her person is guilty of a misdemeanor unless...

  15. The Disillusion of American Dream in Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    张晴

    2016-01-01

    Dreiser is a generally acknowledged writer as one of American's literary naturalists. In Sister Carrie, the author skillfully used the naturalistic writing style which incisively and vividly showed the society environment at that time. The paper states the process that how the American dream in Sister Carrie disillusioned gradually.

  16. [Pathological features of hepatocellular carcinoma carrying hepatitis C virus antigen].

    Science.gov (United States)

    Liu, B; Zhu, S; Zhang, X

    1997-10-01

    To analyze the pathological features of hepatocellular carcinomas (HCCs) carrying different hepatitis virus antigens histopathologically and systematically. PAP and ABC kits were used in the immunohistochemical study, and CAS-200 System was applied in the image cytometry. As compared with HCCs carrying HBV marker(s), the HCCs carrying HCV marker showed more cases of clear cell type (7/9 vs 4/33), better differentiation of cancers, less necrosis of hepatocytes, milder lymphocyte infiltration in the hepatic sinuses or periportal areas (P < 0.01), higher incidence of bile ductule damages, and bore a close relation to the formation of lymphoid follicle in the surrounding tissues (P < 0.05). These patients were elder, with lower grade of symptoms and better prognosis after operation. HCCs carrying only HCAg have different pathological features and clinical characteristics from which carrying HBV marker(s). The results of image cytometry are in accordance with the biological behaviour of HCC.

  17. National attitudes concerning gun carrying in the United States.

    Science.gov (United States)

    Hemenway, D; Azrael, D; Miller, M

    2001-12-01

    To determine public attitudes in the United States concerning gun carrying. In the past 15 years, many state legislatures have passed laws making it easier for United States citizens to carry concealed firearms, not only on the street but into various locations, including churches and government buildings. National random digit dial telephone surveys conducted in 1996 and 1999 asked questions concerning the public's feelings of safety as more people in their community carry firearms, and whether, in the language of the question, respondents believe "regular" citizens should be allowed to carry guns into public or government buildings. Americans feel less safe rather than more safe as more people in their community begin to carry guns. By margins of at least nine to one, Americans do not believe that "regular" citizens should be allowed to bring their guns into restaurants, college campuses, sports stadiums, bars, hospitals, or government buildings. The public believes that increased gun carrying by others reduces rather than increases their safety. Overwhelmingly, the public believes that in many venues gun carrying should be prohibited.

  18. Research on Psychological Carrying Capacity of Tourism Destination

    Institute of Scientific and Technical Information of China (English)

    Fan Zhiyong; Zhong Sheng

    2009-01-01

    As a part of the carrying capacity system of tourism destination,tourism psychological carrying capacity and its makeup are very important indexes which reflect the harmonious development of tourism destination develops harmoniously,but the academy has not paid enough attention to them.Based on the concept and connotation of psychological carrying capacity,this paper explains the influencing factors which affect the psychological capacity of the tourist and the resident after the acknowledged concept,and then designs a harmonious development model of tourism destination.Finally,it offers some countermeasures against the overloading psychological capacity.

  19. Global change and carrying capacity: Implications for life on Earth

    Science.gov (United States)

    Ehrlich, Paul R.; Daily, Gretchen C.; Ehrlich, Anne H.; Matson, Pamela; Vitousek, Peter

    1989-01-01

    Determining the long-term number of people that the planet can support without irreversibly reducing its ability to support people in the future, i.e., the carrying capacity of the Earth, is an exceedingly complex problem. About all that is known for certain is that, with present and foreseeable technologies, the human population has already exceeded the capacity. The reduction in carrying capacity that can be expected to result from direct human impacts on resources and the environment and from our indirect impacts of the climate system is discussed. Global warming and modeling global change and food security are also discussed with respect to carrying capacity.

  20. Global change and carrying capacity: Implications for life on Earth

    Science.gov (United States)

    Ehrlich, Paul R.; Daily, Gretchen C.; Ehrlich, Anne H.; Matson, Pamela; Vitousek, Peter

    1989-01-01

    Determining the long-term number of people that the planet can support without irreversibly reducing its ability to support people in the future, i.e., the carrying capacity of the Earth, is an exceedingly complex problem. About all that is known for certain is that, with present and foreseeable technologies, the human population has already exceeded the capacity. The reduction in carrying capacity that can be expected to result from direct human impacts on resources and the environment and from our indirect impacts of the climate system is discussed. Global warming and modeling global change and food security are also discussed with respect to carrying capacity.

  1. Estimating the recreational carrying capacity of a lowland river section.

    Science.gov (United States)

    Lorenz, Stefan; Pusch, Martin T

    2012-01-01

    Recreational boating represents a major human use of inland waters in many regions. However, boating tourism may affect the ecological integrity of surface waters in multiple ways. In particular, surface waves produced by boating may disturb freshwater invertebrates, such as interrupting the filtration activity of benthic mussels. As mussels may significantly contribute to self-purification, disturbance may have crucial impacts on water quality, and thus on water tourism. In this paper we calculate the carrying capacity of a river section for sustainable boating tourism based on the preservation of water quality. This approach is complemented by spatial and social approaches for carrying capacity estimates. The ecological carrying capacity significantly decreases with lower water levels during summer. Hence, the analysis of variables that influence the river's carrying capacity allows the formation of recommendations for management measures that integrate social, touristic and ecological aspects.

  2. Carrying Network Accessing Architecture and Strategy Based on Business Differentiation

    Directory of Open Access Journals (Sweden)

    Yanyan Han

    2013-07-01

    Full Text Available Due to the abilities of real-time sensing and information sharing, Wireless Sensor Network (WSN has been applied in more and more fields. Basing on the emergence of Internet of Things (IoT, the issue about heterogeneous network integration is becoming more important. We first analyze the new businesses that arise recently for cell phone users as well as the potential effect on carrying network. After that we mainly discuss the influence on traditional carrying network for WSN accessing and taking concurrent businesses as the study case, common access architecture from WSN to carrying network is constructed, which makes use of business differentiation. Furthermore, we propose the idea of tortuous access from WSN to the gateway in the carrying network to avoid congested paths with simulation and verification. Finally, we conclude the possible impacts for the integration of these two networks and present possible solutions.

  3. Lane of parallel through carry in ternary optical adder

    Institute of Scientific and Technical Information of China (English)

    JIN Yi; HE Huacan; AI Lirong

    2005-01-01

    At the present 50 to 100 microseconds are necessary for a liquid crystal to change its state from opacity to clarity; 1.14× 10-5 microseconds are however proved to be enough for light to pass through a clarity liquid crystal device. Rooted from this great difference in time, an optical adder was constructed with parallel through carry lanes (PTCL) composed of liquid crystals. Because all carries in PTCL process in parallel, the carry delay in the ternary optical computer's adder is avoided. Eliminating the carry delay in adder of ternary optical computer by physical means, the PTCL is also applicable for other types of optical adders. Moreover a light diagram of the adder and one PTCL structure are provided.

  4. Study on Tourist Carrying Capacity Based on Matter Element Analysis

    Institute of Scientific and Technical Information of China (English)

    LiuYunguo; FanTing; LiXin; ZhouMing; WangXianhai

    2005-01-01

    This paper proposes that it is necessary to implement the concept of tourist carrying capacity to facilitate the tourism planning, and presents a method to evaluate the carrying capacity. The method called matter element analysis can solve the uncertain and incompatible problem of the evaluated factors in assessing carrying capacity.The current state of a destination's carrying capacity can be determined by establishing the standard indexes and the matter element model. Through the evaluating of the travel industry zones of the Autonomous Prefecture of Western Hunan, the method is proved to be simple and feasible, and it is improved to be significant for the tourism planning and determination as well as the sustainable development of the regional tourism.

  5. Determination of the sediment carrying capacity based on perturbed theory.

    Science.gov (United States)

    Ni, Zhi-hui; Zeng, Qiang; Li-chun, Wu

    2014-01-01

    According to the previous studies of sediment carrying capacity, a new method of sediment carrying capacity on perturbed theory was proposed. By taking into account the average water depth, average flow velocity, settling velocity, and other influencing factors and introducing the median grain size as one main influencing factor in deriving the new formula, we established a new sediment carrying capacity formula. The coefficients were determined by the principle of dimensional analysis, multiple linear regression method, and the least square method. After that, the new formula was verified through measuring data of natural rivers and flume tests and comparing the verified results calculated by Cao Formula, Zhang Formula, Li Formula, Engelung-Hansen Formula, Ackers-White Formula, and Yang Formula. According to the compared results, it can be seen that the new method is of high accuracy. It could be a useful reference for the determination of sediment carrying capacity.

  6. Measuring the Magnetic Force on a Current-Carrying Conductor.

    Science.gov (United States)

    Herreman, W.; Huysentruyt, R.

    1995-01-01

    Describes a fast and simple method for measuring the magnetic force acting on a current-carrying conductor using a digital balance. Discusses the influence of current intensity and wire length on the magnetic force on the conductor. (JRH)

  7. The logistic model-generated carrying capacities for wild herbivores ...

    African Journals Online (AJOL)

    Jesse

    Modelled as discrete-time logistic equations with fixed carrying capacities, it captures the wildlife herbivore population dynamics. Time series data, covering a period ..... Building Models for Conservation and. Wildlife Management. (New York ...

  8. Langkawi Island, Social Aspect and the Carrying Capacity

    OpenAIRE

    Mohamad Diana; Jaafar Mastura; Marzuki Azizan

    2014-01-01

    The aptitude to convince a range of preferences has facilitated the island tourism attractions to achieve international recognition and at present KILIM Geopark is enjoyed by a prominent percentage of individuals wide-reaching. Conventional knowledge has that the island attractions are operating on the limited immobile resources, of which, have raised the scholars’ concerns over the carrying capacity issue. When analyzing carrying capacity, scholars have traditionally focused on the environme...

  9. The Construction of Social Darwinism in Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    薛晶晶; 秦素华

    2014-01-01

    Social Darwinism is an important theory reflected in Sister Carrie. This thesis is to interpret the characters’fates with Social Darwinism.“Survival of the fittest”is an important theme. Carrie is the winner of life and Hurtwood is the loser. Their adjustability to the environment determines their future. It is hoped to help the readers construct a better understanding on this theory.

  10. Design and Implementation of High Speed Carry Select Adder

    OpenAIRE

    B. Gopinath; N Sangeetha; S.Jenifer nancy; T.Umarani

    2015-01-01

    In electronic adder is a digital circuit that performs addition of numbers. Adders can be constructed for many numerical representations such as arithmetic and logical operation. The most adders operates on binary numbers. Among the different types of adders, carry select adder is a one of the fastest adder.The gate level modification is to reduce the power and area of carry select adder by using the concept of Arithmetic Logic Unit (ALU). In this paper , different techniques such as Binary T...

  11. Missile captive carry monitoring using a capacitive MEMS accelerometer

    Science.gov (United States)

    Hatchell, Brian; Mauss, Fred; Santiago-Rojas, Emiliano; Amaya, Ivan; Skorpik, Jim; Silvers, Kurt; Marotta, Steve

    2010-03-01

    Military missiles are exposed to many sources of mechanical vibration that can affect system reliability, safety, and mission effectiveness. One of the most significant exposures to vibration occurs when the missile is being carried by an aviation platform, which is a condition known as captive carry. If the duration of captive carry exposure could be recorded during the missile's service life, several advantages could be realized. Missiles that have been exposed to durations outside the design envelop could be flagged or screened for maintenance or inspection; lightly exposed missiles could be selected for critical mission applications; and missile allocation to missions could be based on prior use to avoid overuse. The U. S. Army Aviation and Missile Research Development and Engineering Center (AMRDEC) has been developing health monitoring systems to assess and improve reliability of missiles during storage and field exposures. Under the direction of AMRDEC staff, engineers at the Pacific Northwest National Laboratory have developed a Captive Carry Health Monitor (CCHM) for the HELLFIRE II missile. The CCHM is an embedded usage monitoring device installed on the outer skin of the HELLFIRE II missile to record the cumulative hours the host missile has been in captive carry mode and thereby assess the overall health of the missile. This paper provides an overview of the CCHM electrical and package design, describes field testing and data analysis techniques used to identify captive carry, and discusses the potential application of missile health and usage data for real-time reliability analysis and fleet management.

  12. Missile Captive Carry Monitoring using a Capacitive MEMS Accelerometer

    Energy Technology Data Exchange (ETDEWEB)

    Hatchell, Brian K.; Mauss, Fredrick J.; Santiago-Rojas, Emiliano; Amaya, Ivan A.; Skorpik, James R.; Silvers, Kurt L.; Marotta, Steve

    2010-04-08

    Military missiles are exposed to many sources of mechanical vibration that can affect system reliability, safety, and mission effectiveness. One of the most significant exposures to vibration occurs when the missile is being carried by an aviation platform, which is a condition known as captive carry. If the duration of captive carry exposure could be recorded during the missile’s service life, several advantages could be realized. Missiles that have been exposed to durations outside the design envelop could be flagged or screened for maintenance or inspection; lightly exposed missiles could be selected for critical mission applications; and missile allocation to missions could be based on prior use to avoid overuse. The U. S. Army Aviation and Missile Research Development and Engineering Center (AMRDEC) has been developing health monitoring systems to assess and improve reliability of missiles during storage and field exposures. Under the direction of AMRDEC staff, engineers at the Pacific Northwest National Laboratory have developed a Captive Carry Health Monitor (CCHM) for the HELLFIRE II missile. The CCHM is an embedded usage monitoring device installed on the outer skin of the HELLFIRE II missile to record the cumulative hours the host missile has been in captive carry mode and thereby assess the overall health of the missile. This paper provides an overview of the CCHM electrical and package design, describes field testing and data analysis techniques used to identify captive carry, and discusses the potential application of missile health and usage data for real-time reliability analysis and fleet management.

  13. Carrying capacity: the tradition and policy implications of limits

    Directory of Open Access Journals (Sweden)

    Virginia Deane Abernethy

    2001-01-01

    Full Text Available ABSTRACT: Within just the last few centuries, science and technology have enlarged human capabilities and population size until humans now take, for their own use, nearly half of the Earth's net terrestrial primary production. An ethical perspective suggests that potentials to alter, or further increase, humanity's use of global resources should be scrutinized through the lenses of self-interested foresightedness and respect for non-human life. Without overtly invoking ethics, studies of the carrying capacity achieve just this objective. Carrying capacity is an ecological concept that expresses the relationship between a population and the natural environment on which it depends for ongoing sustenance. Carrying capacity assumes limits on the number of individuals that can be supported at a given level of consumption without degrading the environment and, therefore, reducing future carrying capacity. That is, carrying capacity addresses long-term sustainability. Worldviews differ in the importance accorded to the carrying capacity concept. This paper addresses three worldviews - ecological, romantic, and entrepreneurial - and explores the ethics and the policy implications of their contrasting perspectives.

  14. Locomotion while load-carrying in reduced gravities.

    Science.gov (United States)

    Wickman, L A; Luna, B

    1996-10-01

    Supporting the mass of a protective suit and portable life support system (PLSS) will impose an energy requirement on planetary astronauts. To design extravehicular protective equipment for planetary missions, scientists must learn more about human physical capabilities while load-carrying in reduced gravities. In this study, an underwater treadmill and weighting system were used to simulate reduced-gravity locomotion while load-carrying. The test matrix included 3 gravity levels, 6 subjects, 2 locomotion speeds, and a range of load sizes. Energy expenditure, calculated from measured oxygen consumption, is positively correlated with gravity level, speed, and load size. The data are used to project that individuals in average physical condition will be able to walk for 8 h on the Moon while carrying up to 170% of their body mass without undue fatigue, and on Mars with up to 50% of their body mass. These approximate limits, especially for Martian gravity, may prove quite a challenge for designers of advanced protective systems. Requirements for regenerable and non-venting PLSS components have been driving the total projected masses of advanced PLSSs increasingly higher, perhaps beyond what is reasonable to carry. However, the larger mass can be beneficial in maintaining bone mass. Using Whalen's model (1988), the daily planetary walking times required to maintain bone mass were calculated for a range of carried load sizes. The calculated times were unattainably high, suggesting that some combination of loads carrying and supplemental bone maintenance measures will likely be required to maintain bone mass in reduced gravity environments.

  15. The Broken American Dream Based on Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    Dong Wenjuan

    2012-01-01

      Sister Carrie is the first work of Theodore Dreiser, It tells us how a country maiden becomes a famous star in Broadway. In the novel, Dreiser describes the living conditions and emotion changes of Carrie very vividly. Sister Carrie takes true realism as its distinctive feature, reflects the tragic fact that people pursue American dream fanatically in the early twentieth century. This paper exposes the theme of instinct, which drives people to enjoy but disillusion finally, and illustrates that it is impossible to possess true happiness in a money-oriented capitalism society%  Sister Carrie is the first work of Theodore Dreiser, Ittells us how a country maiden becomes a famous star in Broadway.In the novel, Dreiser describes the living conditions and emotion changes of Carrie very vividly. Sister Carrie takes true realism as itsdistinctive feature, reflects the tragic fact that people pursue Americandream fanatically in the early twentieth century. This paper exposes the theme of instinct, which drives people to enjoy but disillusion finally, and illustrates that it is impossible to possess true happiness in a money-oriented capitalism society

  16. Integrated Coastal Zone Planning Based on Environment Carrying Capacity Analysis

    Science.gov (United States)

    Miharja, M.; Arsallia, S.

    2017-07-01

    Coastal zone is a crucial area in terms of planning development. It holds high economic value, which affect to increasing number of inhabitants living in the area. As a result, this condition influences environmental degradation. Thus, in every attempt towards coastal zone development, it is crucial to always refer to environment carrying capacity. Carrying capacity is the limit of a certain coastal zone capability to support all human created activities, in which all ecological performances are maintained at sustainable level. The failure to establish strong and clear method and regulation on carrying capacity analysis will lead to a very risky coastal zone development, which in turn would threat the area’s sustainability. This paper discusses method for analysing carrying capacity of coastal zone as important input for the area development plan. Two key parameters, i.e. land and clean water carrying capacities are discussed to form carrying capacity analytical method. Furthermore, an empirical data of Ambon Bay, Moluccas Province, is used to illustrate the operationalization of the method.

  17. Reducing weapon-carrying among urban American Indian young people.

    Science.gov (United States)

    Bearinger, Linda H; Pettingell, Sandra L; Resnick, Michael D; Potthoff, Sandra J

    2010-07-01

    To examine the likelihood of weapon-carrying among urban American Indian young people, given the presence of salient risk and protective factors. The study used data from a confidential, self-report Urban Indian Youth Health Survey with 200 forced-choice items examining risk and protective factors and social, contextual, and demographic information. Between 1995 and 1998, 569 American Indian youths, aged 9-15 years, completed surveys administered in public schools and an after-school program. Using logistic regression, probability profiles compared the likelihood of weapon-carrying, given the combinations of salient risk and protective factors. In the final models, weapon-carrying was associated significantly with one risk factor (substance use) and two protective factors (school connectedness, perceiving peers as having prosocial behavior attitudes/norms). With one risk factor and two protective factors, in various combinations in the models, the likelihood of weapon carrying ranged from 4% (with two protective factors and no risk factor in the model) to 80% of youth (with the risk factor and no protective factors in the model). Even in the presence of the risk factor, the two protective factors decreased the likelihood of weapon-carrying to 25%. This analysis highlights the importance of protective factors in comprehensive assessments and interventions for vulnerable youth. In that the risk factor and two protective factors significantly related to weapon-carrying are amenable to intervention at both individual and population-focused levels, study findings offer a guide for prioritizing strategies for decreasing weapon-carrying among urban American Indian young people. Copyright (c) 2010 Society for Adolescent Health and Medicine. Published by Elsevier Inc. All rights reserved.

  18. Bullying and weapon carrying: a meta-analysis.

    Science.gov (United States)

    van Geel, Mitch; Vedder, Paul; Tanilon, Jenny

    2014-08-01

    Studies suggest that adolescents involved in bullying are more likely to carry weapons than their uninvolved peers. To use meta-analyses to determine whether victims, bullies, and bully-victims are more likely to carry weapons than uninvolved peers. PsycINFO, ERIC, MEDLINE, LILACS, EMBASE, and Dissertation Abstracts International were searched for relevant publications (1950 through January 2014). The reference list of a review article and reference lists of retrieved articles were checked for further relevant studies. Studies were included if they provided an effect size comparing the weapon carrying of adolescent victims, bullies, or bully-victims with that of uninvolved peers. Studies that included individuals older than 21 years were excluded, as were studies that focused on incarcerated youth or youth diagnosed as having a psychopathologic condition. Studies were coded independently by 2 of us. The agreement rate was 93%. Effect sizes were coded that compared victims, bullies, or bully-victims with uninvolved peers. Meta-analyses were based on 22 studies for victims (n = 257 179), 15 studies for bullies (n = 236 145), and 8 studies for bully-victims (n = 199 563). This study focused on weapon carrying among adolescents. Hypotheses were formulated before the study. Victims (odds ratio, 1.97; 95% CI, 1.62-2.39), bullies (3.25; 2.72-3.89), and bully-victims (4.95; 3.77-6.50) were more likely to carry weapons than uninvolved peers. Analyses provided no indication of publication bias. Studies conducted in the United States found stronger relations between being a bully-victim and weapon carrying (odds ratio, 7.84; 95% CI, 6.02-10.21) than studies from other countries (3.62; 2.30-5.68; Q1 = 8.401; P = .004). Involvement in bullying as a victim, bully, or bully-victim is related to weapon carrying.

  19. Model Modification and Application on Carrying Capacity of Relative Resources

    Institute of Scientific and Technical Information of China (English)

    Li Zehong; Dong Suocheng; Gao Dan

    2009-01-01

    Based on the retrospection of researches on carry-ing capacity, this article reviewed systematically the research progresses on carrying capacity of relative resources (CCRR).Then the viewpoint was put forward that CCRR is not an ap-propriate method of appraising the regional sustainability, but a sound way to obtain cognition for coordinating spatial loca-tion and flow of population and economy.However, as the most popular computing method of CCRR, the Weighting Linear Sum Model is defective in the random of weight choice and the neglect of matching among different resources.Therefore, this article established the Geometric Model on CCRR based on modifying Weighting Linear Sum Model, which can be used to appraise regions where resources are close matching.Em-ploying the Geometric Model, the article empirically analyzed the population and economic CCRR in Hubei Province from 1978 to 2006.The result indicates that the population in Hubei Province is overloading while the economic carrying capacity is abundant compared to the whole country, and the economic insufficiency restricts the population carrying capacity.In the future, Hubei Province will become one of the core developing zones which are characterized by economic conglomeration.

  20. Boundary slippage for generating hydrodynamic load-carrying capacity

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yong-bin

    2008-01-01

    Boundary slippage is used to generate the load-carrying capacity of the hydrodynamic contact between two parallel plane surfaces.In the fluid inlet zone,the fluidcontact interfacial shear strength on a stationary surface is set at low to generate boundary slippage there,while in the fluid outlet zone the fluid-contact interfacial shear strength on the stationary surface is set at high enough to prevent the occurrence of boundary slippage.The fluid-contact interfacial shear strength on the entire moving surface is set at high enough to prevent boundary slippage on the moving surface.These hydrodynamic contact configurations are analyzed to generate the pronounced load-carrying capacity.The optimum ratio of the outlet zone width to the inlet zone width for the maximum load-carrying capacity of the whole contact is found to be 0.5.

  1. Hierarchical population model with a carrying capacity distribution

    CERN Document Server

    Indekeu, J O

    2002-01-01

    A time- and space-discrete model for the growth of a rapidly saturating local biological population $N(x,t)$ is derived from a hierarchical random deposition process previously studied in statistical physics. Two biologically relevant parameters, the probabilities of birth, $B$, and of death, $D$, determine the carrying capacity $K$. Due to the randomness the population depends strongly on position, $x$, and there is a distribution of carrying capacities, $\\Pi (K)$. This distribution has self-similar character owing to the imposed hierarchy. The most probable carrying capacity and its probability are studied as a function of $B$ and $D$. The effective growth rate decreases with time, roughly as in a Verhulst process. The model is possibly applicable, for example, to bacteria forming a "towering pillar" biofilm. The bacteria divide on randomly distributed nutrient-rich regions and are exposed to random local bactericidal agent (antibiotic spray). A gradual overall temperature change away from optimal growth co...

  2. Evaluation of carrying capacity and territorial environmental sustainability

    Directory of Open Access Journals (Sweden)

    Giuseppe Ruggiero

    2012-09-01

    Full Text Available Land use has a great impact on environmental quality, use of resources, state of ecosystems and socio-economic development. Land use can be considered sustainable if the environmental pressures of human activities do not exceed the ecological carrying capacity. A scientific knowledge of the capability of ecosystems to provide resources and absorb waste is a useful and innovative means of supporting territorial planning. This study examines the area of the Province of Bari to estimate the ecosystems’ carrying capacity, and compare it with the current environmental pressures exerted by human activities. The adapted methodology identified the environmentally sustainable level for one province.

  3. Design and Analysis of a High Speed Carry Select Adder

    OpenAIRE

    Simarpreet Singh Chawla; Swapnil Aggarwal; Anshika; Nidhi Goel

    2015-01-01

    An optimal high-speed and low-power VLSI architecture requires an efficient arithmetic processing unit that is optimized for speed and power consumption. Adders are one of the widely used in digital integrated circuit and system design.High speed adder is the necessary component in a data path, e.g. Microprocessors and a Digital signal processor. The present paper proposes a novel high-speed adder by combining the advantages of Carry Look Ahead Adder (CLAA) and Carry Select Adder (CSA), devis...

  4. Design and Analysis of a High Speed Carry Select Adder

    OpenAIRE

    Simarpreet Singh Chawla; Swapnil Aggarwal; Anshika; Nidhi Goel

    2015-01-01

    An optimal high-speed and low-power VLSI architecture requires an efficient arithmetic processing unit that is optimized for speed and power consumption. Adders are one of the widely used in digital integrated circuit and system design. High speed adder is the necessary component in a data path, e.g. Microprocessors and a Digital signal processor. The present paper proposes a novel high-speed adder by combining the advantages of Carry Look Ahead Adder (CLAA) and Carry Select Adder (CSA), devi...

  5. Description of Model Tests Carried Out by Aalborg University

    DEFF Research Database (Denmark)

    Frigaard, Peter; Schlütter, F.; Andersen, H.

    1996-01-01

    As associated partner, Aalborg University (AU) have participated in different aspects of "the Zeebrugge project". AU has carried out an extensive number of small-scale model tests (1:65) with the Zeebrugge breakwater with the aim of investigating scale-effects.......As associated partner, Aalborg University (AU) have participated in different aspects of "the Zeebrugge project". AU has carried out an extensive number of small-scale model tests (1:65) with the Zeebrugge breakwater with the aim of investigating scale-effects....

  6. Ticks Carrying Lyme Disease Confirmed in Eastern National Parks

    Science.gov (United States)

    ... html Ticks Carrying Lyme Disease Confirmed in Eastern National Parks U.S. National Park Service and CDC advise using insect repellents on ... Planning a hiking trip in an eastern U.S. national park? Better pack tick repellent -- a new study found ...

  7. 44 CFR 332.3 - Carrying out voluntary agreements.

    Science.gov (United States)

    2010-10-01

    ... advance, each meeting of the participants in the agreement held to discuss problems, determine policies... § 332.5 of this part. A person attending a meeting under this section may present oral or written data... communications among themselves or with any other member of their industry, related to the carrying out of...

  8. El naturalismo americano: Theodore Dreiser y Sister Carrie

    Directory of Open Access Journals (Sweden)

    Dolores G. ALONSO MULAS

    2009-08-01

    Full Text Available Para situar a un escritor, como Theodore Dreiser, y especialmente su novela Sister Carrie dentro de un movimiento literario y de una etapa determinada de la historia americana, es necesario dar un breve repaso al naturalismo, llegado a América a través de Stephen Crane

  9. Carrying Out Collaborative Action Research in a Practicum

    Science.gov (United States)

    Ho, Belinda

    2013-01-01

    To help in-service English teachers improve their teaching practices and pupils' learning opportunities through an enquiry-oriented and cooperative approach, the author carried out collaborative action research during a practicum in a teacher education course at a university in Hong Kong. This article describes how the collaborative research…

  10. Arrangement for carrying out and studying chemical reactions

    NARCIS (Netherlands)

    Langeveld, A.D.; Makkee, M.; Moulijn, J.A.

    1993-01-01

    Abstract of NL 9102029 (A) Described is an arrangement for carrying out TAP experiments. Such an arrangement comprises a reaction chamber in which a very short-lived beam of molecules is directed at a target to produce a short-lived beam of intermediate-product molecules, and a vacuum chamber wher

  11. Victimization and Health Risk Factors among Weapon-Carrying Youth

    Science.gov (United States)

    Stayton, Catherine; McVeigh, Katharine H.; Olson, E. Carolyn; Perkins, Krystal; Kerker, Bonnie D.

    2011-01-01

    Objective: To compare health risks of 2 subgroups of weapon carriers: victimized and nonvictimized youth. Methods: 2003-2007 NYC Youth Risk Behavior Surveys were analyzed using bivariate analyses and multinomial logistic regression. Results: Among NYC teens, 7.5% reported weapon carrying without victimization; 6.9% reported it with victimization.…

  12. Ferromagnetic behavior of formyl-group-carrying stable thioaminyl radicals.

    Science.gov (United States)

    Miura, Yozo; Nakamura, Shogo; Teki, Yoshio

    2003-10-17

    Four formyl-group-carrying thioaminyl radicals were generated, and one radical could be isolated as radical crystals. Magnetic susceptibility measurements of the isolated radical showed a ferromagnetic regular linear-chain interaction of 2J/k(B) = 3.2 K, which was explained in terms of the X-ray crystallographic results.

  13. Debris-carrying camouflage among diverse lineages of Cretaceous insects

    Science.gov (United States)

    Wang, Bo; Xia, Fangyuan; Engel, Michael S.; Perrichot, Vincent; Shi, Gongle; Zhang, Haichun; Chen, Jun; Jarzembowski, Edmund A.; Wappler, Torsten; Rust, Jes

    2016-01-01

    Insects have evolved diverse methods of camouflage that have played an important role in their evolutionary success. Debris-carrying, a behavior of actively harvesting and carrying exogenous materials, is among the most fascinating and complex behaviors because it requires not only an ability to recognize, collect, and carry materials but also evolutionary adaptations in related morphological characteristics. However, the fossil record of such behavior is extremely scarce, and only a single Mesozoic example from Spanish amber has been recorded; therefore, little is known about the early evolution of this complicated behavior and its underlying anatomy. We report a diverse insect assemblage of exceptionally preserved debris carriers from Cretaceous Burmese, French, and Lebanese ambers, including the earliest known chrysopoid larvae (green lacewings), myrmeleontoid larvae (split-footed lacewings and owlflies), and reduviids (assassin bugs). These ancient insects used a variety of debris material, including insect exoskeletons, sand grains, soil dust, leaf trichomes of gleicheniacean ferns, wood fibers, and other vegetal debris. They convergently evolved their debris-carrying behavior through multiple pathways, which expressed a high degree of evolutionary plasticity. We demonstrate that the behavioral repertoire, which is associated with considerable morphological adaptations, was already widespread among insects by at least the Mid-Cretaceous. Together with the previously known Spanish specimen, these fossils are the oldest direct evidence of camouflaging behavior in the fossil record. Our findings provide a novel insight into early evolution of camouflage in insects and ancient ecological associations among plants and insects. PMID:27386568

  14. Cultural Carrying Capacity: A Biological Approach to Human Problems.

    Science.gov (United States)

    Hardin, Garrett

    1992-01-01

    In discussing the human and cultural implications of scientific discoveries and knowledge, the biological concept of carrying capacity is explored. Maintaining that human beings are truly animals answering to principles that govern all animals, the author addresses the need for human populations to work within the context of culture and carrying…

  15. Cognitive Load in Voice Therapy Carry-Over Exercises

    Science.gov (United States)

    Iwarsson, Jenny; Morris, David Jackson; Balling, Laura Winther

    2017-01-01

    Purpose: The cognitive load generated by online speech production may vary with the nature of the speech task. This article examines 3 speech tasks used in voice therapy carry-over exercises, in which a patient is required to adopt and automatize new voice behaviors, ultimately in daily spontaneous communication. Method: Twelve subjects produced…

  16. Wakes in Dilatonic Current-Carrying Cosmic Strings

    CERN Document Server

    Oliveira, A L N

    2003-01-01

    In this work, we present the gravitational field generated by a cosmic string carrying a timelike current in the scalar-tensor gravities. The mechanism of formation and evolution of wakes is fully investigated in this framework. We show that the inclusion of electromagnetic properties for the string induces logarithmic divergences in the accretion problem.

  17. Nanoparticles carry chemotherapy drug deeper into solid tumors

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    @@ CAS researchers have developed a new drug delivery method using nano-sized molecules to carry the chemotherapy drug doxorubicin to tumors, improving the effectiveness of the drug in mice and increasing their survival time. Their work has been reported online June 26 in the Journal of the National Cancer Institute.

  18. Maze Busters: Carrie Miyoshi Macfarlane & Kathleen Sheehan--Harvard University

    Science.gov (United States)

    Library Journal, 2004

    2004-01-01

    Even if one is equipped with an MLS, the 11 libraries that comprise the Harvard College Library can be pretty daunting. That is why Carrie Miyoshi Macfarlane and Kathleen Sheehan created Threading the Maze. The online publication is presented to students in expository writing, the one course all undergraduates must take. "This highly effective…

  19. Connection and Commitment: The Career of Carrie Billy

    Science.gov (United States)

    Peterson, Richard

    2015-01-01

    Carrie Billy, a member of the Navajo Nation and CEO and president of the American Indian Higher Education Consortium (AIHEC), attributes one of the country's oldest youth organizations, Girl Scouts, for opening her mind to the possibility of creating change for countless citizens. She is a trained attorney and has made it her life work to advocate…

  20. "The Bell Curve" and Carrie Buck: Eugenics Revisited.

    Science.gov (United States)

    Smith, J. David

    1995-01-01

    The 1994 publication of "The Bell Curve" by R. Herrnstein and C. Murray is compared to other examples of eugenic principles, including the sterilization of "feebleminded" Carrie Buck, family degeneracy studies focusing on lower class Caucasian families, and other works that view the poorest and least educated members of society…

  1. Pyrosequencing reveals bacteria carried in different wind eroded sediments

    Science.gov (United States)

    Little is known about the microbial communities carried in wind-eroded sediments from various soil types and land management systems. A novel technique, pyrosequencing, promises to expand our understanding of the vast microbial diversity of soils and eroded sediments as it can sequence between 10-10...

  2. Efficient Design of Ripple Carry Adder and Carry Skip Adder with Low Quantum Cost and Low Power Consumption

    Directory of Open Access Journals (Sweden)

    Nidhi

    2014-07-01

    Full Text Available The addition of two binary numbers is the important and most frequently used arithmetic process on microprocessors, digital signal processors (DSP, and data-processing application-specific integrated circuits (ASIC. Therefore, binary adders are critical structure blocks in very large-scale integrated (VLSI circuits. Their effective application is not trivial because a costly carry spread operation involving all operand bits has to be achieved. Many different circuit constructions for binary addition have been planned over the last decades, covering a wide range of presentation characteristics. In today era, reversibility has become essential part of digital world to make digital circuits more efficient. In this paper, we have proposed a new method to reduce quantum cost for ripple carry adder and carry skip adder. The results are simulated in Xilinx by using VHDL language.

  3. Variable Block Carry Skip Logic using Reversible Gates

    CERN Document Server

    Islam, Md Rafiqul; Karim, Muhammad Rezaul; Mahmud, Abdullah Al; Babu, Hafiz Md Hasan

    2010-01-01

    Reversible circuits have applications in digital signal processing, computer graphics, quantum computation and cryptography. In this paper, a generalized k*k reversible gate family is proposed and a 3*3 gate of the family is discussed. Inverter, AND, OR, NAND, NOR, and EXOR gates can be realized by this gate. Implementation of a full-adder circuit using two such 3*3 gates is given. This full-adder circuit contains only two reversible gates and produces no extra garbage outputs. The proposed full-adder circuit is efficient in terms of gate count, garbage outputs and quantum cost. A 4-bit carry skip adder is designed using this full-adder circuit and a variable block carry skip adder is discussed. Necessary equations required to evaluate these adder are presented.

  4. High Speed Boosted Cmos Differential Logic for Ripple Carry Adders

    Directory of Open Access Journals (Sweden)

    Meenu Roy,

    2014-01-01

    Full Text Available This paper describes a high speed boosted CMOS differential logic which is applicable in Ripple Carry Adders. The proposed logic operating with supply voltage approaching the MOS threshold voltage. The logic style improves switching speed by boosting the gate-source voltage of transistors along timing critical signal path. It allows a single boosting circuit to be shared by complementary outputs as a result the area overhead also minimizes. As compared to the conventional logic gates the EDP (energy delay product is improved. The test sets of logic gates and adders where designed in tsmc0.18μm of Mentor Graphics EDA tool. The experimental result for Ripple Carry Adders using the proposed logic style revealed that the addition time is reduced as compared with the conventional CMOS circuits.

  5. A novel reversible carry-selected adder with low latency

    Science.gov (United States)

    Li, Ming-Cui; Zhou, Ri-Gui

    2016-07-01

    Reversible logic is getting more and more attention in quantum computing, optical computing, nanotechnology and low-power complementary metal oxide semiconductor designs since reversible circuits do not loose information during computation and have only small energy dissipation. In this paper, a novel carry-selected reversible adder is proposed primarily optimised for low latency. A 4-bit reversible full adder with two kinds of outputs, minimum delay and optimal quantum cost is presented as the building block for ?-bit reversible adder. Three new reversible gates NPG (new Peres gate), TEPG (triple extension of Peres gate) and RMUX21 (reversible 2-to-1 multiplexer) are proposed and utilised to design efficient adder units. The secondary carry propagation chain is carefully designed to reduce the time consumption. The novelty of the proposed design is the consideration of low latency. The comparative study shows that the proposed adder achieves the improvement from 61.46% to 95.29% in delay over the existing designs.

  6. Carrying guns in public: legal and public health implications.

    Science.gov (United States)

    Vernick, Jon S

    2013-03-01

    In District of Columbia v. Heller, the U.S. Supreme Court ruled that the Second Amendment protects an individual's right to own handguns in the home for protection, invalidating a Washington, D.C. law banning most handgun possession. The Heller decision, however, provided lower courts with little guidance regarding how to judge the constitutionality of gun laws other than handgun bans. Nevertheless, lower courts have upheld the vast majority of federal, state, and local gun laws challenged since Heller. One area in which some lower courts have disagreed has been the constitutionality of laws regulating the ability to carry firearms in public. This issue may be the next to be addressed by the Supreme Court under its evolving Second Amendment jurisprudence. Courts should carefully consider the negative public health and safety implications of gun carrying in public as they weigh the constitutionality of these laws.

  7. A C-Testable Multiple-Block Carry Select Adder

    Science.gov (United States)

    Kito, Nobutaka; Fujii, Shinichi; Takagi, Naofumi

    We propose a C-testable multiple-block carry select adder with respect to the cell fault model. Full adders and 2: 1 multiplexers are considered as cells. By an additional external input, we obtain a C-testable carry select adder. We only modify the least significant position of each block. The adder is testable with a test set consisting of 16 patterns regardless of the size of each block and the number of blocks. This is the minimum test set for the adder. We show two gate-level implementations of the adder which are testable with a test set of 9 patterns and 7 patterns respectively, with respect to the single stuck-at fault model.

  8. Different Ways of Carrying Bags in Mental Retarded Students

    Directory of Open Access Journals (Sweden)

    Saideh-Sadat Mortazavi

    2012-10-01

    Full Text Available School as an educational and social center is linked with various aspects of child’s growth. The impact of objects, tools, and lifestyle on the development of human performance is not hidden from anyone [1].Tasell Lauri, the head of Cairo Praktive in Australia, in a paper warned parents about bags as a tool that students carry every day, and he is believes that many spinal disorders in adults is caused by childhood trauma. Studies on children and patients with special needs and such as myelomenangocele, cerebral palsy, and neurofibromatosis precisely show abnormal growth of spinal disorders, muscle imbalance, wrong posture [2]. Carrying stationery in an inappropriate tool can cause irreversible effects.

  9. Superpositions of light fields carrying orbital angular momentum

    CSIR Research Space (South Africa)

    Dudley, Angela L

    2012-01-01

    Full Text Available OF LIGHT FIELDS CARRYING ORBITAL ANGULAR MOMENTUM By Angela Dudley A thesis submitted in fulfilment of the academic requirements for the PhD degree of Science in the School of Chemistry and Physics, University of KwaZulu-Natal, Durban... this dissertation for submission. _________________________________ Prof Andrew Forbes On this_______day of____________________________2012 iv Declaration 2 - Plagiarism I, ???????????????. declare that 1...

  10. Modeling the Flexural Carrying Capacity of Corroded RC Beam

    Institute of Scientific and Technical Information of China (English)

    WANG Xiao-hui; LIU Xi-la

    2008-01-01

    Considering the change of bond strength between corroded steel and concrete, flexural carrying ca-pacity of corroded reinforced concrete (RC) beam was calculated. On the basis of the condition of equilibriumof forces and compatibility of deformations for the whole beam, a model for the prediction of flexural carryingcapacity of the corroded RC beam was proposed. Comparison of the model's predictions with the experimentalresults published in the literature shows the practicality of the proposed method.

  11. The calming effect of maternal carrying in different mammalian species

    Directory of Open Access Journals (Sweden)

    Gianluca eEsposito

    2015-04-01

    Full Text Available Attachment theory postulates that mothers and their infants possess some basic physiological mechanisms that favour their dyadic interaction and bonding. Many studies have focused on the maternal physiological mechanisms that promote attachment (e.g. mothers’ automatic responses to infant faces and/or cries, and relatively less have examined infant physiology. Thus, the physiological mechanisms regulating infant bonding behaviors remain largely undefined. This review elucidates some of the neurobiological mechanisms governing social bonding and cooperation in humans by focusing on maternal carrying and its beneficial effect on mother-infant interaction in mammalian species (e.g. in humans, big cats and rodents. These studies show that infants have a specific calming response to maternal carrying. A human infant carried by his/ her walking mother exhibits a rapid heart rate decrease, and immediately stops voluntary movement and crying compared to when he/ she is held in a sitting position. Furthermore, strikingly similar responses were identified in mouse rodents, who exhibit immobility, diminished ultra-sonic vocalizations and heart rate. In general, the studies described in the current review demonstrate the calming effect of maternal carrying to be comprised of a complex set of behavioral and physiological components, each of which has a specific postnatal time window and is orchestrated in a well-matched manner with the maturation of the infants. Such reactions could have been evolutionarily adaptive in mammalian mother-infant interactions. The findings have implications for parenting practices in developmentally normal populations. In addition, we propose that infants’ physiological response may be useful in clinical assessments as we discuss possible implications on early screening for child psychopathology (e.g. Autism Spectrum Disorders, and Perinatal Brain Disorders.

  12. The calming effect of maternal carrying in different mammalian species.

    Science.gov (United States)

    Esposito, Gianluca; Setoh, Peipei; Yoshida, Sachine; Kuroda, Kumi O

    2015-01-01

    Attachment theory postulates that mothers and their infants possess some basic physiological mechanisms that favor their dyadic interaction and bonding. Many studies have focused on the maternal physiological mechanisms that promote attachment (e.g., mothers' automatic responses to infant faces and/or cries), and relatively less have examined infant physiology. Thus, the physiological mechanisms regulating infant bonding behaviors remain largely undefined. This review elucidates some of the neurobiological mechanisms governing social bonding and cooperation in humans by focusing on maternal carrying and its beneficial effect on mother-infant interaction in mammalian species (e.g., in humans, big cats, and rodents). These studies show that infants have a specific calming response to maternal carrying. A human infant carried by his/her walking mother exhibits a rapid heart rate decrease, and immediately stops voluntary movement and crying compared to when he/she is held in a sitting position. Furthermore, strikingly similar responses were identified in mouse rodents, who exhibit immobility, diminished ultra-sonic vocalizations and heart rate. In general, the studies described in the current review demonstrate the calming effect of maternal carrying to be comprised of a complex set of behavioral and physiological components, each of which has a specific postnatal time window and is orchestrated in a well-matched manner with the maturation of the infants. Such reactions could have been evolutionarily adaptive in mammalian mother-infant interactions. The findings have implications for parenting practices in developmentally normal populations. In addition, we propose that infants' physiological response may be useful in clinical assessments as we discuss possible implications on early screening for child psychopathology (e.g., autism spectrum disorders and perinatal brain disorders).

  13. High Speed Carry Select Adder for ALU Blocks

    Directory of Open Access Journals (Sweden)

    J.Rama Krishna Reddy

    2013-06-01

    Full Text Available The regular SQRT CSLA consists of two RCA blocks with carry input as 0 and 1.The Final sum will be selected from multiplexers (Mux by the carry out generated by the pervious block. This paper, proposes an area and delay efficient carry select adder with logical reduction of excess redundant hardware. In the proposed architecture, we had implemented the RCA with carry input as 1, only with Mux, Or gate and And gate. For 16-bit regular SQRT CSLA there is a reduction of basic logic gates from 434 to 323.The delay is reduced by replacing Full-adder with half-adder in first bit of every RCA in theproposed architecture. This will reduces the number of Iterations required to get the final sum. The proposed architecture shows that there is reduction of area and delay. Based on this architecture, wedesigned 4-bit,8-bit,16-bit and 32-bit Square-root CSLA (SQRT CSLA and compared with the regular SQRT CSLA. In this work, we evaluated the performance of the proposed design in 90-ηm CMOS Technology in Cadence Tools. The result analysis shows that, the proposed SQRT CSLA of 4-bit, 8-bit 16- bit and 32-bit has a reduction of 31.74%, 30.13%, 21.92% and 21.76 % respectively compared with regular SQRT CSLA in area. The delay of Proposed SQRT CSLA of 4-bit,8-bit 16-bit and 32-bit are reduce by 27.47%, 17.23%, 14.32% and 11.63% respectively.

  14. Gluteus medius and thigh muscles electromyography during load carrying walking

    Directory of Open Access Journals (Sweden)

    Petr Šťastný

    2015-11-01

    Full Text Available This study compares the electromyographic (EMG peak amplitude changes of gluteus medius (Gmed, vastus medialis (VMO, vastus lateralis (VL and biceps femoris (BF during load carrying walking due to the increased load. The percentage of maximum isometric voluntary contractions (%MVIC of both limbs and 3D kinematic of lower limbs were detected on eighteen resistance-trained men (mean age ± SD, 31 ± 3.4 years while carrying loads of 25, 50 and 75% of their body mass (BM. The repeated measurement ANOVA was used to evaluate the differences in muscles %MVIC and 3D kinematics at all load conditions. Significant differences were found for Gmed %MVIC (F3,99 = 19.8, p < 0.001. Gmed activity was significantly different between load carrying walking with 25% of BM (mean ± SD, 20 ± 12%MVIC, 50% of BM (32 ± 17%MVIC and 75% of BM (45 ± 26%MVIC condition. Differences were found in hip flexion at Gmed EMG peak (F3,96 = 14, p < 0.001, between 25% of BM (18 ± 11° and 50% of BM (29 ± 7°. No significant differences were found for thigh muscles, when thigh muscle activity did not exceed 30%MVIC even at 75% of BM condition. Load carrying walking is an exercise which activates Gmed more than thigh muscles. This exercise increases the Gmed activity along with increased loads and it should be regarded as a complex Gmed strengthening exercise. This exercise is recommended for strengthening the Gmed with low activation of VL and VMO.

  15. Self-timed Manchester chain carry propagate adder

    OpenAIRE

    Escribà, J; Carrasco, Juan A.

    1996-01-01

    The authors present a self-timed adder that uses two Manchester chains to propagate carries in a two-rail code. With the inclusion of buffers in the chains, the adder meets the timing conditions typical of an asynchronous design based in the ‘bundled-data, bounded-delay’ model and is signifcantly faster than self-timed adders with restoring logic and similar complexity.

  16. Low Power 256-bit Modified Carry Select Adder

    Directory of Open Access Journals (Sweden)

    P. Ramani

    2014-09-01

    Full Text Available Carry Select Adder (CSLA is one of the high speed adders used in many computational systems to perform fast arithmetic operations. When compared to earlier Ripple Carry Adder and Carry Look Ahead Adder, Regular CSLA (R-CSLA is observed to provide optimized results in terms of area. This study proposes an efficient method which replaces the RCA using BEC. The modified CSLA architecture has been developed using gate-level modification to significantly reduce the delay and power of the CSLA. Based on this modification 8-, 16-, 32-, 64- and 128-bit Square-Root CSLA (SQRT CSLA architecture have been developed and compared with the regular SQRT CSLA architecture. The proposed design for 256-bit has reduced power and delay as compared with the regular SQRT CSLA. Designs were developed using structural Verilog module and synthesized using Xilinx ISE simulator and the implementation is done in cadence RTL compiler using 0.18 µm technology. For 256-bit addition in this study, it is proposed to simple gate level modification which significantly reduces the power by 19.4% when compared with R-CSLA. The result analysis shows that the proposed architecture achieves two folded advantages in terms of delay and power.

  17. A portable virtual machine target for proof-carrying code

    DEFF Research Database (Denmark)

    Franz, Michael; Chandra, Deepak; Gal, Andreas

    2005-01-01

    Virtual Machines (VMs) and Proof-Carrying Code (PCC) are two techniques that have been used independently to provide safety for (mobile) code. Existing virtual machines, such as the Java VM, have several drawbacks: First, the effort required for safety verification is considerable. Second and mor...... simultaneously providing efficient justin-time compilation and target-machine independence. In particular, our approach reduces the complexity of the required proofs, resulting in fewer proof obligations that need to be discharged at the target machine.......Virtual Machines (VMs) and Proof-Carrying Code (PCC) are two techniques that have been used independently to provide safety for (mobile) code. Existing virtual machines, such as the Java VM, have several drawbacks: First, the effort required for safety verification is considerable. Second and more...... subtly, the need to provide such verification by the code consumer inhibits the amount of optimization that can be performed by the code producer. This in turn makes justin-time compilation surprisingly expensive. Proof-Carrying Code, on the other hand, has its own set of limitations, among which...

  18. Targeted drug-carrying bacteriophages as antibacterial nanomedicines.

    Science.gov (United States)

    Yacoby, Iftach; Bar, Hagit; Benhar, Itai

    2007-06-01

    While the resistance of bacteria to traditional antibiotics is a major public health concern, the use of extremely potent antibacterial agents is limited by their lack of selectivity. As in cancer therapy, antibacterial targeted therapy could provide an opportunity to reintroduce toxic substances to the antibacterial arsenal. A desirable targeted antibacterial agent should combine binding specificity, a large drug payload per binding event, and a programmed drug release mechanism. Recently, we presented a novel application of filamentous bacteriophages as targeted drug carriers that could partially inhibit the growth of Staphylococcus aureus bacteria. This partial success was due to limitations of drug-loading capacity that resulted from the hydrophobicity of the drug. Here we present a novel drug conjugation chemistry which is based on connecting hydrophobic drugs to the phage via aminoglycoside antibiotics that serve as solubility-enhancing branched linkers. This new formulation allowed a significantly larger drug-carrying capacity of the phages, resulting in a drastic improvement in their performance as targeted drug-carrying nanoparticles. As an example for a potential systemic use for potent agents that are limited for topical use, we present antibody-targeted phage nanoparticles that carry a large payload of the hemolytic antibiotic chloramphenicol connected through the aminoglycoside neomycin. We demonstrate complete growth inhibition toward the pathogens Staphylococcus aureus, Streptococcus pyogenes, and Escherichia coli with an improvement in potency by a factor of approximately 20,000 compared to the free drug.

  19. Violence and weapon carrying in music videos. A content analysis.

    Science.gov (United States)

    DuRant, R H; Rich, M; Emans, S J; Rome, E S; Allred, E; Woods, E R

    1997-05-01

    The positive portrayal of violence and weapon carrying in televised music videos is thought to have a considerable influence on the normative expectations of adolescents about these behaviors. To perform a content analysis of the depictions of violence and weapon carrying in music videos, including 5 genres of music (rock, rap, adult contemporary, rhythm and blues, and country), from 4 television networks and to analyze the degree of sexuality or eroticism portrayed in each video and its association with violence and weapon carrying, as an indicator of the desirability of violent behaviors. Five hundred eighteen videos were recorded during randomly selected days and times of the day from the Music Television, Video Hits One, Black Entertainment Television, and Country Music Television networks. Four female and 4 male observers aged 17 to 24 years were trained to use a standardized content analysis instrument. Interobserver reliability testing resulted in a mean (+/- SD) percentage agreement of 89.25% +/- 7.10% and a mean (+/- SD) kappa of 0.73 +/- 0.20. All videos were observed by rotating 2-person, male-female teams that were required to reach agreement on each behavior that was scored. Music genre and network differences in behaviors were analyzed with chi 2 tests. A higher percentage (22.4%) of Music Television videos portrayed overt violence than Video Hits One (11.8%), Country Music Television (11.8%), and Black Entertainment Television (11.5%) videos (P = .02). Rap (20.4%) had the highest portrayal of violence, followed by rock (19.8%), country (10.8%), adult contemporary (9.7%), and rhythm and blues (5.9%) (P = .006). Weapon carrying was higher on Music Television (25.0%) than on Black Entertainment Television (11.5%), Video Hits One (8.4%), and Country Music Television (6.9%) (P rhythm and blues (6.9%), and country (6.3%) videos (P = .002). The videos with the highest level of sexuality or eroticism were found to be less likely to contain violence (P < or

  20. Efficient Design of Ripple Carry Adder and Carry Skip Adder with Low Quantum Cost and Low Power Consumption

    OpenAIRE

    Nidhi; Gurinderpal Singh

    2014-01-01

    The addition of two binary numbers is the important and most frequently used arithmetic process on microprocessors, digital signal processors (DSP), and data-processing application-specific integrated circuits (ASIC). Therefore, binary adders are critical structure blocks in very large-scale integrated (VLSI) circuits. Their effective application is not trivial because a costly carry spread operation involving all operand bits has to be achieved. Many different circuit constru...

  1. Killing cancer cells by targeted drug-carrying phage nanomedicines

    Directory of Open Access Journals (Sweden)

    Yacoby Iftach

    2008-04-01

    Full Text Available Abstract Background Systemic administration of chemotherapeutic agents, in addition to its anti-tumor benefits, results in indiscriminate drug distribution and severe toxicity. This shortcoming may be overcome by targeted drug-carrying platforms that ferry the drug to the tumor site while limiting exposure to non-target tissues and organs. Results We present a new form of targeted anti-cancer therapy in the form of targeted drug-carrying phage nanoparticles. Our approach is based on genetically-modified and chemically manipulated filamentous bacteriophages. The genetic manipulation endows the phages with the ability to display a host-specificity-conferring ligand. The phages are loaded with a large payload of a cytotoxic drug by chemical conjugation. In the presented examples we used anti ErbB2 and anti ERGR antibodies as targeting moieties, the drug hygromycin conjugated to the phages by a covalent amide bond, or the drug doxorubicin conjugated to genetically-engineered cathepsin-B sites on the phage coat. We show that targeting of phage nanomedicines via specific antibodies to receptors on cancer cell membranes results in endocytosis, intracellular degradation, and drug release, resulting in growth inhibition of the target cells in vitro with a potentiation factor of >1000 over the corresponding free drugs. Conclusion The results of the proof-of concept study presented here reveal important features regarding the potential of filamentous phages to serve as drug-delivery platform, on the affect of drug solubility or hydrophobicity on the target specificity of the platform and on the effect of drug release mechanism on the potency of the platform. These results define targeted drug-carrying filamentous phage nanoparticles as a unique type of antibody-drug conjugates.

  2. [Simple locomotion and during load carrying in pregnant women].

    Science.gov (United States)

    Golomer, E; Ducher, D; Arfi, G S; Sud, R

    1991-01-01

    There have been few studies of the way women walk in pregnancy, though some of the causes of low back ache of which they complain have only been partially worked out. This is why this study has been carried out on ten women between the third and eighth month of pregnancy. The speed at which they walk and the parameters of the gait as well as the localization of the centre of gravity when keeping upright have been measured in these pregnant women as well as in twenty control women of the same age. The results show that the speed at which they walk whether with or without carrying a weight usually is identical at the beginning and the end of pregnancy. When walking normally the size of the steps taken are no larger in pregnant women than in the control patients (p less than 0.05). Though the results are not statistically significant the rhythm of the steps is faster as well as their being a reduction in the length of the steps between the third and eighth month of pregnancy. When carrying a weight the length of the steps does not change greatly with pregnancy (p less than 0.05) and it would seem therefore to be a good way of assessing the changes that locomotion undergoes in pregnancy. The fact that women do not walk faster or slower can give evidence that they adapt to the change in posture that happens in pregnancy and they make the best possible biomechanical use of the parameters of walking in order to economise total energy output of the organism.(ABSTRACT TRUNCATED AT 250 WORDS)

  3. Design and Verification of Advanced Carry Select Adder

    Directory of Open Access Journals (Sweden)

    Guru Dixit Chepuri

    2014-05-01

    Full Text Available Design ofareaefficient data pathlogicsystems formsthelargestareasofresearch inVLSIsystem design.Indigital adders, thespeed of addition islimitedbythetimerequired totransmit a carrythroughtheadder.CarrySelectAdder (CSA isoneofthefastestadders usedinmanydata-processingprocessorstoperform fastarithmetic functions. Fromthestructure oftheCSA,itisclear thatthereisscopeforreducing thearea and powerintheCSA. This work usesasimpleandefficient gate-level modification todrasticallyreducetheareaandpower oftheCSA.Basedonthismodification 16,32,64 and128-bitsquare-root CSA(SQRT CSA architectureshavebeendevelopedandcomparedwith theregularSQRTCSAarchitecture. Theproposed designhasreducedarea and powerascompared withtheregular SQRTCSA.Thisworkestimates theperformanceof theproposed designsintermsofpower,areaand isimplementedusingXilinxISE and synthesized using cadence in 90nm technology.

  4. Optimal Final Carry Propagate Adder Design for Parallel Multipliers

    CERN Document Server

    B., Ramkumar

    2011-01-01

    Based on the ASIC layout level simulation of 7 types of adder structures each of four different sizes, i.e. a total of 28 adders, we propose expressions for the width of each of the three regions of the final Carry Propagate Adder (CPA) to be used in parallel multipliers. We also propose the types of adders to be used in each region that would lead to the optimal performance of the hybrid final adders in parallel multipliers. This work evaluates the complete performance of the analyzed designs in terms of delay, area, power through custom design and layout in 0.18 um CMOS process technology.

  5. Reflection of Naturalism and Darwinism in Dreiser' s Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    王静

    2009-01-01

    Naturalism is a significant school of literature in American literary history. This thesis mainly reviews the background, development, and characteristic of the realism and naturalism literature, and states the significant person in Naturalism literature-Dreiser and his. important literary position and analyzes the reflection of naturalism and Darwinism in Sister Carrie. Through the analysis of determinism, desire, ethics and detail description, a conclusion is made: the factors affecting the novel' s writing not only include the author' s own experiences but also the main social ideology in his living years.

  6. The information carrying capacity of a cosmological constant

    CERN Document Server

    Simidzija, Petar

    2016-01-01

    We analyze the exchange of information in different cosmological backgrounds when sender and receiver are timelike separated and communicate through massless fields (without the exchange of light-signals). Remarkably, we show that the dominance of a cosmological constant makes the amount of recoverable information imprinted in the field by the sender extremely resilient: it does not decay in time or with the spatial separation of sender and receiver, and it actually increases with the rate of expansion of the Universe. This is in stark contrast with the information carried by conventional light-signals and with previous results on timelike communication through massless fields in matter dominated cosmologies.

  7. The carry-over effect does not influence football results.

    OpenAIRE

    Goossens, Dries; Spieksma, Frederik

    2012-01-01

    In a round robin tournament, it is often believed that each team has an effect on its opponent, which carries over to the next game of that opponent. Indeed, if team A plays against team B, and subsequently against team C, A’s performance against C may have been affected by B, and we say that team C receives a carryover effect from B. For instance, if team B is a very strong team, then team A could be exhausted and discouraged after this game, which could benefit its next opponent, team C. Cl...

  8. Langkawi Island, Social Aspect and the Carrying Capacity

    Directory of Open Access Journals (Sweden)

    Mohamad Diana

    2014-01-01

    Full Text Available The aptitude to convince a range of preferences has facilitated the island tourism attractions to achieve international recognition and at present KILIM Geopark is enjoyed by a prominent percentage of individuals wide-reaching. Conventional knowledge has that the island attractions are operating on the limited immobile resources, of which, have raised the scholars’ concerns over the carrying capacity issue. When analyzing carrying capacity, scholars have traditionally focused on the environmental part, hence, this working paper is motivated to convey onto the table issues relating to the social characteristics. This particular paper employed the self-administered questionnaire survey instrument which was structured to answer the two-fold objective specifically the tourists’ satisfaction level with their tourism experience and knowledge gap in relation to improving the island tourism. Targeting the tourists with minimum age of 18, this working paper collects information on the tourists’ perception towards hospitality, facilities and safety issues. In addition, this working paper comes to scrutinize different results of demographic factors as compared to what have been documented by earlier studies. Besides, it is learned that ‘safety issue’ variable plays an important role when it comes to the following factors: tourist arrival, access to facilities and tourism experience.

  9. Are food brands that carry light claims different?

    DEFF Research Database (Denmark)

    Sjostrom, Therese; Corsi, Armando Maria; Driesener, Carl

    2014-01-01

    Little is known about the market performance of brands that carry light claims (e.g. low fat, low sugar) in comparison to their regular counterparts. In order to fill this gap, we explore whether light brands perform similarly to regular brands in terms of a) brand performance measures, such as m......Little is known about the market performance of brands that carry light claims (e.g. low fat, low sugar) in comparison to their regular counterparts. In order to fill this gap, we explore whether light brands perform similarly to regular brands in terms of a) brand performance measures......, such as market share and penetration, b) loyalty levels, and c) customer sharing. We analyse three product categories (Cola, Flavoured Carbonated Beverages and Margarine) using UK household panel data provided by Kantar. The results show that when considering standard brand performance measures (i.e. market...... share, penetration and purchase frequency), regular brands receive higher brand performance measures than light brands. However, when considering repeat purchase loyalty, light brands achieve greater levels of loyalty than their regular counterparts. Finally, light brands share their buyers more...

  10. Kinematic analysis of males with transtibial amputation carrying military loads

    Directory of Open Access Journals (Sweden)

    Barri L. Schnall, MPT

    2015-03-01

    Full Text Available The biomechanical responses to load carriage, a common task for dismounted troops, have been well studied in nondisabled individuals. However, with recent shifts in the rehabilitation and retention process of injured servicemembers, there remains a substantial need for understanding these responses in persons with lower-limb amputations. Temporal-spatial and kinematic gait parameters were analyzed among 10 male servicemembers with unilateral transtibial amputation (TTA and 10 uninjured male controls. Participants completed six treadmill walking trials in all combinations of two speeds (1.34 and 1.52 m/s and three loads (none, 21.8, and 32.7 kg. Persons with TTA exhibited biomechanical compensations to carried loads that are comparable to those observed in uninjured individuals. However, several distinct gait changes appear to be unique to those with TTA, notably, increased dorsiflexion (deformation of the prosthetic foot/ankle, less stance knee flexion on the prosthetic limb, and altered trunk forward lean/excursion. Such evidence supports the need for future work to assess the risk for overuse injuries with carried loads in this population in addition to guiding the development of adaptive prosthetic feet/components to meet the needs of redeployed servicemembers or veterans/civilians in physically demanding occupations.

  11. c-Myc-Induced Extrachromosomal Elements Carry Active Chromatin

    Directory of Open Access Journals (Sweden)

    Greg Smith

    2003-03-01

    Full Text Available Murine Pre-13 lymphocytes with experimentally activated MycER show both chromosomal and extrachromosomal gene amplification. In this report, we have elucidated the size, structure, functional components of c-Myc-induced extrachromosomal elements (EEs. Scanning electron microscopy revealed that EEs isolated from MycER-activated Pre-B+ cells are an average of 10 times larger than EEs isolated from non-MycER-activated control Pre-B- cells. We demonstrate that these large c-Myc-induced EEs are associated with histone proteins, whereas EEs of non-MycER-activated Pre B- cells are not. Immunohistochemistry and Western blot analyses using pan -histone-specific, histone H3 phosphorylation-specific, histone H4 acetylation-specific antibodies indicate that a significant proportion of EEs analyzed from MycER-activated cells harbors transcriptionally competent and/or active chromatin. Moreover, these large, c-Myc-induced EEs carry genes. Whereas the total genetic make-up of these c-Myc-induced EEs is unknown, we found that 30.2% of them contain the dihydrofolate reductase (DHFR gene, whereas cyclin C (CCNC was absent. In addition, 50% of these c-Myc-activated Pre-B+ EEs incorporated bromodeoxyuridine (BrdU, identifying them as genetic structures that self-propagate. In contrast, EEs isolated from non-Myc-activated cells neither carry the DHFR gene nor incorporate BrdU, suggesting that c-Myc deregulation generates a new class of EEs.

  12. The Population Carrying Capacity of Water Resources in Yulin City

    Institute of Scientific and Technical Information of China (English)

    Lijuan; DANG; Yong; XU; Zhiqiang; WANG

    2014-01-01

    Assessing the water resource carrying capacity is beneficial for measuring the scale of industry and population agglomeration,and also avoiding the contradiction between increasing people and decreasing available water resource,due to the expansion of industry and city size.Based on the prediction model of optimum population development size,by using hydrological data,also with the demographic data from 1956 to 2010,this article analyzes and predicts the urban moderate scale under the limit of the water resource in the future of Yulin City by GIS. The main conclusions are as follows. There is growing tendency of water resources overloading. According to the result of model simulation,by2015,the overload rate of population size will be 1. 04. By 2020,the overload rate of population size will grow up to 1. 08. The oversized population mainly comes from cities and towns. The overload rate for cities and towns in 2015 and 2020 is 1. 89 and 1. 73,respectively. With the expansion of cities and industries,suburban areas could have a great potential for carrying population,because lots of suburban people may move to cities and towns according to prediction. In view of the above-mentioned facts,the population size should be controlled in a reasonable range.

  13. A Novel Framework to Carry Out Cloud Penetration Test

    Directory of Open Access Journals (Sweden)

    Jianbin Hu

    2011-04-01

    Full Text Available in current cloud services, users put their data and resources into the cloud so as to enjoy the on-demand high quality applications and services. Different from the conventional services, users in cloud services lose control of their data which is instead manipulated by the large-scale cloud. Therefore, cloud service providers (CSP guarantee that the cloud which they provide is of high confidence in accuracy and integrity. Traditional penetration test is carried out manually and has low efficiency. In this paper, we propose FPTC, a novel framework of penetration test in cloud environment. In FPTC, there are managers, executors and toolkits. FPTC managers guide FPTC executors to gather information from the cloud environment, generate appropriate testing scenarios, run matched tools in the toolkit and collect test results to do evaluation. The capacity and quality of the toolkit is a key issue in FPTC. We develop a prototype in which FPTC is implemented and the experimental results show that FPTC is helpful to automatically carry out penetration test in cloud environment.

  14. Modelling of carry-over in recovery furnaces

    Energy Technology Data Exchange (ETDEWEB)

    Fakhrai, Reza [Royal Inst. of Tech., Stockholm (Sweden). Dept. of Metallurgy

    2000-04-01

    Development of mathematical modelling of the combustion process in the furnace of recovery boilers is the subject of this work. This work as a continuation of many years of modelling efforts carried out at KTH/Vaerme- och Ugnsteknik focussed particularly on: char bed modelling; droplets-wall interaction modelling; and carry-over modelling. The char bed model has been studied. Droplets/parcels were considered as a single reactor working independently of the other droplets. The mass of the droplets was not distributed uniformly but induced in the landing place. The droplets hitting the char bed will stick to it and they are alive and part of the calculation. In this way the distribution of the mass on the char bed is only dependent on the parameters which effect flight history such as droplet/parcel diameter, boilers flow field, etc. The droplet- wall interaction model has been studied and found to be very important for obtaining the correct temperature distribution in the recovery furnace. The new approach is based on removal of droplets which hits the wall in the upper part of the recovery boiler from carryover calculation. This model has been proposed and implemented into the GRFM (General Recovery Furnace Model). The carryover modelling effort was based on mass balance in which the number and physical statistics of the droplets/parcel were estimated and the amount of unburned mass was calculated. All of the above listed models were tested together with all other models of heat and mass transfer processes in recovery furnaces using a GRFM. Three-dimensional numerical simulations of the industrial recovery boiler (63 kg/s, 82 bar, 480 deg C) were performed. The number of grid was 232,000 and the number of air ports in this simulation was 178. The air entering the furnace by these ports has different flow rates. Flow and temperature fields as well as species distributions were calculated. The results show good agreement with previously published data and modelling

  15. Carrying capacity in agriculture: environmental significance and some related patents.

    Science.gov (United States)

    da Silva, Alexandre M

    2009-06-01

    Agriculture is one of the most important and possibly the oldest economic activity developed by humans. This activity was developed extensively and is becoming more and more dependent on development of technologies. The goal of this manuscript was examining some patents related to technologies developed for improving crop yields. Such patents are mainly related to more efficient formulations of agrochemicals and management techniques of plants, cattle and natural resources. A brief comment is carried out about bioprospection and related problems, relating, for example the case of Cupuaçu. The article is concluded mentioning that the development of policies and management strategies that increase agricultural yield and simultaneously preserve or conserve natural resources should also be prioritized, because certainly this is the only way we have to get the real sustainability and to improve life quality abroad the world.

  16. Carbohydrate nanotechnology: hierarchical assembly using nature's other information carrying biopolymers.

    Science.gov (United States)

    Han, Xu; Zheng, Yeting; Munro, Catherine J; Ji, Yiwen; Braunschweig, Adam B

    2015-08-01

    Despite their central role in directing some of the most complex biological processes, carbohydrates--nature's other information carrying biopolymer--have been largely ignored as building blocks for synthetic hierarchical assemblies. The non-stoichiometric binding and astronomical diversity characteristic of carbohydrates could lead to tantalizingly complex assembly algorithms, but these attributes simultaneously increase the difficulty of preparing carbohydrate assemblies and anticipating their behavior. Convergences in biotechnology, nanotechnology, polymer chemistry, surface science, and supramolecular chemistry have led to many recent important breakthroughs in glycan microarrays and synthetic carbohydrate receptors, where the idiosyncrasies of carbohydrate structure and binding are increasingly considered. We hope to inspire more researchers to consider carbohydrate structure, diversity, and binding as attractive tools for constructing synthetic hierarchical assemblies.

  17. Cognitive Load in Voice Therapy Carry-Over Exercises

    DEFF Research Database (Denmark)

    Iwarsson, Jenny; Morris, David Jackson; Balling, Laura Winther

    2017-01-01

    Purpose The cognitive load generated by online speech production may vary with the nature of the speech task. This article examines 3 speech tasks used in voice therapy carry-over exercises, in which a patient is required to adopt and automatize new voice behaviors, ultimately in daily spontaneous...... communication. Method Twelve subjects produced speech in 3 conditions: rote speech (weekdays), sentences in a set form, and semispontaneous speech. Subjects simultaneously performed a secondary visual discrimination task for which response times were measured. On completion of each speech task, subjects rated...... their experience on a questionnaire. Results Response times from the secondary, visual task were found to be shortest for the rote speech, longer for the semispontaneous speech, and longest for the sentences within the set framework. Principal components derived from the subjective ratings were found to be linked...

  18. Pallet Optimization of the Heavy Rotary Table Load Carrying System

    Science.gov (United States)

    Atapin, V. G.; Bataev, A. A.

    2016-04-01

    The pallet optimization of the heavy rotary table load-carrying system, which is a part of the multi-purpose machine, is considered in terms of the deterministic and probabilistic models. As a result of optimum design in case of the deterministic model the mass of the pallet is reduced by 35.5 % in comparison with a serial model. The evaluation of the influence of optimization problem limitations on design variables confirms the importance of rigidity criterion in relation to other criteria. Calculation for probabilistic model allows reducing the mass of the construction by 27 % in comparison with the deterministic model. Considering a work piece rigidity on the basis of a conventional work piece of the minimum rigidity (without stiffening ribs etc.) leads to reducing of the pallet mass by 22.3 % in comparison with the deterministic model.

  19. Modeling symbiosis by interactions through species carrying capacities

    Science.gov (United States)

    Yukalov, V. I.; Yukalova, E. P.; Sornette, D.

    2012-08-01

    We introduce a mathematical model of symbiosis between different species by taking into account the influence of each species on the carrying capacities of the others. The modeled entities can pertain to biological and ecological societies or to social, economic and financial societies. Our model includes three basic types: symbiosis with direct mutual interactions, symbiosis with asymmetric interactions, and symbiosis without direct interactions. In all cases, we provide a complete classification of all admissible dynamical regimes. The proposed model of symbiosis turned out to be very rich, as it exhibits four qualitatively different regimes: convergence to stationary states, unbounded exponential growth, finite-time singularity, and finite-time death or extinction of species.

  20. Carry Trade e Risco Cambial: um Conto de Dois Fatores

    Directory of Open Access Journals (Sweden)

    Alex Luiz Ferreira

    2015-12-01

    Full Text Available Retornos da estratégia de carry trade tem sido explicados usando-se funções de utilidade inseparáveis no tempo que permitem prêmios de risco voláteis. Tipicamente tais funções mimetizam as preferências de economia fechada que dependem de bens duráveis e não duráveis. Este trabalho retorna a uma classificação mais tradicional, em macroeconomia internacional, de consumo entre bens domésticos e importados. O modelo é aplicado para países que representam 98,2% do volume mundial do comércio bilateral de câmbio. Reporta-se uma melhora acentuada na significância dos betas de consumo.

  1. Filaggrin compound heterozygous patients carry mutations in trans position

    DEFF Research Database (Denmark)

    Carlsen, Berit C; Meldgaard, Michael; Johansen, Jeanne D

    2013-01-01

    ; however, this has not been scientifically investigated. Two different FLG null mutations in the same individual may be in trans position, meaning that each mutation locates to a different allele functionally equivalent to homozygosity, or may be in cis position, meaning that both mutations locate...... to the same allele functionally equivalent to heterozygosity. To experimentally investigate allelic in cis versus in trans configuration of the two most common filaggrin (FLG) mutations (R501X and 2282del4) in compound heterozygous individuals. Testing for in cis or in trans allele configuration was performed...... compound heterozygous individuals were found to carry the two mutations in trans position. FLG null mutation compound heterozygous individuals can be considered functionally equivalent to FLG null mutation homozygosity for any of the two mutations....

  2. Fatigue Analysis of Load-Carrying Fillet Welds

    DEFF Research Database (Denmark)

    Sørensen, John Dalsgaard; Tychsen, Jesper; Andersen, Jens Ulfkjær

    2006-01-01

    is a doubler plate connection, which is often applied in connection with modifications of offshore structures. As a part of the present work, fatigue tests have been performed with test specimens fabricated by the current industry standard for welded offshore steel structures. The fatigue tests show......The fatigue strength of load-carrying fillet welds is, in most codes of practice, performed neglecting the influence of bending in the weld throat section. However, some commonly applied structural details give rise to significant bending in the weld throat section. An example of such a detail...... that the degree of bending (DOB) has an influence on the fatigue lifetime. The fatigue lifetime decreases significantly when increasing the bending stress. In order to take into account the effect of the bending, a new fatigue stress definition applicable for fillet welds failing through the weld is presented...

  3. Overview of the activities carried out at the FEBEX site

    Energy Technology Data Exchange (ETDEWEB)

    Missana, T.; Buil, B.; Garralon, A.; Gomez, P. [CIEMAT, Dept. de Medioambien te, 28040 Madrid (Spain); Perez-Estaun, A.; Carbonell, R. [Inst. Jaume Almera, CSIC (Spain); Suso, J.; Carretero, G.; Bueno, J.; Martinez, L. [AITEMIN (Spain) ; Hernan, P. [ENRESA (Spain)

    2007-06-15

    One of the main aim of WP 4.1 and 4.2 is to study solute migration mechanisms in crystalline host-rock in realistic conditions. Many organisations are participating in a joint study that is being performed in the FEBEX gallery (NAGRA's Grimsel Test Site, GTS, Switzerland). The FEBEX experiment reproduces at a real scale a high-level waste repository in granite and was installed more than 9 years ago. At moment, it represents the most realistic environment where the processes affecting radionuclide migration from the bentonite to granite can be studied. This paper summarises the main activities carried out at the FEBEX site during the second year of the project.

  4. Rise-Time Distortion of Signal without Carrying Signal

    Science.gov (United States)

    Bukhman, N. S.

    2016-08-01

    The article deals with one-dimensional problem of rise-time distortion signal without carrying signal, that appears in the starting point intermittently, that is signal distortion at front edge or one of its derivative. The authors show that front edge of signal isn't distorted in case of propagation in unrestricted (including absorbing) area (amplitude of starting signal step or of one of its derivatives doesn't change) and move with the accuracy of vacuum light speed. The paper proves that it is the time interval shortage that causes signal loss with the route extension, but not the reduction of its starting amplitude, during which front edge of signal retains its starting value. The research presents new values for this time interval.

  5. Bartonella-like bacteria carried by domestic mite species.

    Science.gov (United States)

    Kopecký, Jan; Nesvorná, Marta; Hubert, Jan

    2014-01-01

    Bacteria of the genus Bartonella are carried by haematophagous mites, ticks, fleas and flies, and attack the erythrocytes of mammals. Here we describe a Bartonella-like clade, a distinct group related to Bartonellaceae, in stored-product mites (Acari: Astigmata) and a predatory mite Cheyletus eruditus (Acari: Prostigmata) based on the analysis of cloned 16S rRNA gene sequences. By using the clade-specific primers, closely related Bartonella-like 16S rRNA sequences were amplified from both laboratory colonies and field strains of three synanthropic mite species (Acarus siro, Lepidoglyphus destructor and Tyrophagus putrescentiae) and a predatory mite. Altogether, sequences of Bartonella-like bacteria were found in 11 strains, but were not detected in Dermatophagoides farinae and D. pteronyssinus and two strains of L. destructor. All obtained sequences formed a separate cluster branching as a sister group to Bartonellaceae and related to other separate clusters comprising uncultured bacterial clones from human skin and hemipteran insects (Nysius plebeius and Nysius sp.). The classification of sequences into operational taxonomic units (OTUs) showed a difference between A. siro and T. putrescentiae suggesting that the Bartonella-like bacteria are different in these two mite species. However, species specific sequences in separate OTUs were observed also for C. eruditus. Possible symbiotic interactions between Bartonella-like bacteria and their mite hosts are discussed.

  6. Saliva microbiota carry caries-specific functional gene signatures.

    Directory of Open Access Journals (Sweden)

    Fang Yang

    Full Text Available Human saliva microbiota is phylogenetically divergent among host individuals yet their roles in health and disease are poorly appreciated. We employed a microbial functional gene microarray, HuMiChip 1.0, to reconstruct the global functional profiles of human saliva microbiota from ten healthy and ten caries-active adults. Saliva microbiota in the pilot population featured a vast diversity of functional genes. No significant distinction in gene number or diversity indices was observed between healthy and caries-active microbiota. However, co-presence network analysis of functional genes revealed that caries-active microbiota was more divergent in non-core genes than healthy microbiota, despite both groups exhibited a similar degree of conservation at their respective core genes. Furthermore, functional gene structure of saliva microbiota could potentially distinguish caries-active patients from healthy hosts. Microbial functions such as Diaminopimelate epimerase, Prephenate dehydrogenase, Pyruvate-formate lyase and N-acetylmuramoyl-L-alanine amidase were significantly linked to caries. Therefore, saliva microbiota carried disease-associated functional signatures, which could be potentially exploited for caries diagnosis.

  7. Certificate size reduction in Abstraction-Carrying Code

    CERN Document Server

    Albert, Elvira; Puebla, Germán; Hermenegildo, Manuel

    2010-01-01

    Carrying Code (ACC) has recently been proposed as a framework for mobile code safety in which the code supplier provides a program together with an abstraction (or abstract model of the program) whose validity entails compliance with a predefined safety policy. The advantage of providing a (fixpoint) abstraction to the code consumer is that its validity is checked in a single pass (i.e., one iteration) of an abstract interpretation-based checker. A main challenge to make ACC useful in practice is to reduce the size of certificates as much as possible while at the same time not increasing checking time. The intuitive idea is to only include in the certificate information that the checker is unable to reproduce without iterating. We introduce the notion of reduced certificate which characterizes the subset of the abstraction which a checker needs in order to validate (and re-construct) the full certificate in a single pass. Based on this notion, we instrument a generic analysis algorithm with the necessary exte...

  8. Carrying photosynthesis genes increases ecological fitness of cyanophage in silico.

    Science.gov (United States)

    Hellweger, Ferdi L

    2009-06-01

    Several viruses infecting marine cyanobacteria carry photosynthesis genes (e.g. psbA, hli) that are expressed, yield proteins (D1, HLIP) and help maintain the cell's photosynthesis apparatus during the latent period. This increases energy and speeds up virus production, allowing for a reduced latent period (a fitness benefit), but it also increases the DNA size, which slows down new virus production and reduces burst size (a fitness cost). How do these genes affect the net ecological fitness of the virus? Here, this question is explored using a combined systems biology and systems ecology ('systems bioecology') approach. A novel agent-based model simulates individual cyanobacteria cells and virus particles, each with their own genes, transcripts, proteins and other properties. The effect of D1 and HLIP proteins is explicitly considered using a mechanistic photosynthesis component. The model is calibrated to the available database for Prochlorococcus ecotype MED4 and podovirus P-SSP7. Laboratory- and field-scale in silico survival, competition and evolution (gene packaging error) experiments with wild type and genetically engineered viruses are performed to develop vertical survival and fitness profiles, and to determine the optimal gene content. The results suggest that photosynthesis genes are nonessential, increase fitness in a manner correlated with irradiance, and that the wild type has an optimal gene content.

  9. Report of mecC-carrying MRSA in domestic swine

    Science.gov (United States)

    Angen, Ø.; Stegger, M.; Larsen, J.; Lilje, B.; Kaya, H.; Pedersen, K. S.; Jakobsen, A.; Petersen, A.; Larsen, A. R.

    2017-01-01

    Objectives We unexpectedly identified MRSA isolates carrying mecC (mecC-MRSA) from a Danish swine farm located in eastern Zealand. The objective of the present study was to investigate the origin of these isolates and their genetic relatedness to other mecC-MRSA isolates from Zealand. Methods WGS was used to infer the phylogenetic relationship between 19 identified mecC-MRSA isolates from the swine farm and 34 additional epidemiologically unrelated human isolates from the same geographical region of Denmark. Variations in the accessory genome were investigated by bioinformatics tools, and antibiotic susceptibility profiles were assessed by MIC determination. Results mecC-MRSA was isolated from a domestic swine farm, but not from cattle reared at the same farm. Phylogenetic analysis revealed that all mecC-MRSA isolates from both farm animals and workers formed a separate cluster, whereas human isolates from the same municipality belonged to a closely related cluster. Analysis of the accessory genome supported this relationship. Conclusions To the best of our knowledge, this is the first report of mecC-MRSA isolated from domestic swine. The investigation strongly indicates that transmission of mecC-MRSA has taken place on the swine farm between the farmers and swine. The close clustering of farm isolates and isolates from the same municipality suggests a local transmission of mecC-MRSA. PMID:27650187

  10. Radar imaging using electromagnetic wave carrying orbital angular momentum

    Science.gov (United States)

    Yuan, Tiezhu; Cheng, Yongqiang; Wang, Hongqiang; Qin, Yuliang; Fan, Bo

    2017-03-01

    The concept of radar imaging based on orbital angular momentum (OAM) modulation, which has the ability of azimuthal resolution without relative motion, has recently been proposed. We investigate this imaging technique further in greater detail. We first analyze the principle of the technique, accounting for its resolving ability physically. The phase and intensity distributions of the OAM-carrying fields produced by phased uniform circular array antenna, which have significant effects on the imaging results, are investigated. The imaging model shows that the received signal has the form of inverse discrete Fourier transform with the use of OAM and frequency diversities. The two-dimensional Fourier transform is employed to reconstruct the target images in the case of large and small elevation angles. Due to the peculiar phase and intensity characteristics, the small elevation is more suitable for practical application than the large one. The minimum elevation angle is then obtained given the array parameters. The imaging capability is analyzed by means of the point spread function. All results are verified through numerical simulations. The proposed staring imaging technique can achieve extremely high azimuthal resolution with the use of plentiful OAM modes.

  11. Wildlife carrying capacities in relation to human settlement

    Directory of Open Access Journals (Sweden)

    S.K. Eltringham

    1990-09-01

    Full Text Available Human encroachment into wildlife areas, which has increased almost exponentially over the past few decades, has usually resulted in the elimination of the larger species, particularly the large mammals. This is not an inevitable consequence and this paper considers the extent to which man and wildlife can coexist. There is a linear inverse relationship between human and elephant densities and the reasons for this are discussed with particular reference to Uganda. Such a relationship does not necessarily hold for all species and the outcome of increasing human pressure on wildlife habitats varies with a variety of factors including the species concerned, the rainfall, vegetation, soil and, above all, the attitudes of the people towards wildlife. Wild animals are more likely to be tolerated if they do no harm to human activities or if the harm they do is outweighed by the benefits to be obtained from their exploitation. In many parts of Africa utilisation is likely to be the best hope for the conservation of wildlife. Some examples are given of situations in which worthwhile carrying capacities of wildlife can be maintained in the presence of human activities.

  12. Human embryonic stem cells carrying mutations for severe genetic disorders.

    Science.gov (United States)

    Frumkin, Tsvia; Malcov, Mira; Telias, Michael; Gold, Veronica; Schwartz, Tamar; Azem, Foad; Amit, Ami; Yaron, Yuval; Ben-Yosef, Dalit

    2010-04-01

    Human embryonic stem cells (HESCs) carrying specific mutations potentially provide a valuable tool for studying genetic disorders in humans. One preferable approach for obtaining these cell lines is by deriving them from affected preimplantation genetically diagnosed embryos. These unique cells are especially important for modeling human genetic disorders for which there are no adequate research models. They can be further used to gain new insights into developmentally regulated events that occur during human embryo development and that are responsible for the manifestation of genetically inherited disorders. They also have great value for the exploration of new therapeutic protocols, including gene-therapy-based treatments and disease-oriented drug screening and discovery. Here, we report the establishment of 15 different mutant human embryonic stem cell lines derived from genetically affected embryos, all donated by couples undergoing preimplantation genetic diagnosis in our in vitro fertilization unit. For further information regarding access to HESC lines from our repository, for research purposes, please email dalitb@tasmc.health.gov.il.

  13. Inverse PCR for subtyping of Acinetobacter baumannii carrying ISAba1.

    Science.gov (United States)

    Kim, Shukho; Park, Yun-Ju; Kim, Jungmin

    2016-05-01

    Acinetobacter baumannii has been prevalent in nosocomial infections, often causing outbreaks in intensive care units. ISAba1 is an insertion sequence that has been identified only in A. baumannii and its copy number varies among strains. It has been reported that ISAba1 provides a promoter for bla(OXA-51-like), bla(OXA-23-like), and bla(ampC), which are associated with the resistance of A. baumannii to carbapenems and cephalosporins. The main purpose of this study was to develop a novel inverse PCR method capable of typing A. baumannii strains. The method involves three major steps: cutting of genomic DNA with a restriction enzyme, ligation, and PCR. In the first step, bacterial genomic DNA was digested with DpnI. In the second step, the digested genomic DNAs were ligated to form intramolecular circular DNAs. In the last step, the ligated circular DNAs were amplified by PCR with primers specific for ISAba1 and the amplified PCR products were electrophoresed. Twenty-two clinical isolates of A. baumannii were used for the evaluation of the inverse PCR (iPCR) typing method. Dendrogram analysis revealed two major clusters, similar to pulsed-field gel electrophoresis (PFGE) results. Three ISAba1-associated genes--bla(ampC), bla(OXA-66-like), and csuD--were amplified and detected in the clinical isolates. This novel iPCR typing method is comparable to PFGE in its ability to discriminate A. baumannii strains, and is a promising molecular epidemiological tool for investigating A. baumannii carrying ISAba1.

  14. Roadmap and performance carried out during Ciemat site decommissioning

    Energy Technology Data Exchange (ETDEWEB)

    Quinones, Javier; Diaz Diaz, Jose Luis

    2005-01-01

    Ciemat (Research Centre for Energy, Environment and Technology) located in the heart of the Ciudad Universitaria of Madrid, occupies a property of 20 Ha. Since its creation in 1951 as JEN, and in 1986 renowned as Ciemat, it has involved on R and D projects in the field of Energy and Environment, i.e., Nuclear Fission, Nuclear Fusion, Fossils Fuels, Renewable Energy. As a consequence of the R and D projects developed between 1951 - 1986 on Nuclear Fission field (fuel design, fabrication, characterization on irradiated fuels, safety studies, etc) and to the diversification of the goals as well, it is necessary to Decommissioning and Dismantling (D and D) from nuclear facilities (nuclear reactor, Hot Cells, Irradiation facility), buildings and soils. Preparations for D and D included a staged shutdown of operations, planning documentation and licensing for decommissioning. As a prerequisite to Ciemat application for a decommissioning license and nuclear environmental assessment was carried out according to Spanish Nuclear Council (CSN) and approval of the site decommissioning project was obtained in 2000 and valid until December 31, 2006. Since 2001 - 2003 is underway and focussed on the radiological characterization of the site (divided in pieces of ground), when each piece of ground is characterized a planning for D and D is presented to CSN in order to obtain a license for actuation. Nowadays several pieces of ground are decontaminated and modifications have been done in order to achieve a safe state of storage-with-surveillance. Later phases have planned waste management improvements for selected wastes already on temporally storage, eventually followed by final decommissioning of facilities and buildings and cleaning of contaminants from soils and removal of waste from the site. This paper describes the planning, nuclear and environment assessment and descriptions of decommissioning activities currently underway at Ciemat. (Author)

  15. Estimating landscape carrying capacity through maximum clique analysis.

    Science.gov (United States)

    Donovan, Therese M; Warrington, Gregory S; Schwenk, W Scott; Dinitz, Jeffrey H

    2012-12-01

    Habitat suitability (HS) maps are widely used tools in wildlife science and establish a link between wildlife populations and landscape pattern. Although HS maps spatially depict the distribution of optimal resources for a species, they do not reveal the population size a landscape is capable of supporting--information that is often crucial for decision makers and managers. We used a new approach, "maximum clique analysis," to demonstrate how HS maps for territorial species can be used to estimate the carrying capacity, N(k), of a given landscape. We estimated the N(k) of Ovenbirds (Seiurus aurocapillus) and bobcats (Lynx rufus) in an 1153-km2 study area in Vermont, USA. These two species were selected to highlight different approaches in building an HS map as well as computational challenges that can arise in a maximum clique analysis. We derived 30-m2 HS maps for each species via occupancy modeling (Ovenbird) and by resource utilization modeling (bobcats). For each species, we then identified all pixel locations on the map (points) that had sufficient resources in the surrounding area to maintain a home range (termed a "pseudo-home range"). These locations were converted to a mathematical graph, where any two points were linked if two pseudo-home ranges could exist on the landscape without violating territory boundaries. We used the program Cliquer to find the maximum clique of each graph. The resulting estimates of N(k) = 236 Ovenbirds and N(k) = 42 female bobcats were sensitive to different assumptions and model inputs. Estimates of N(k) via alternative, ad hoc methods were 1.4 to > 30 times greater than the maximum clique estimate, suggesting that the alternative results may be upwardly biased. The maximum clique analysis was computationally intensive but could handle problems with < 1500 total pseudo-home ranges (points). Given present computational constraints, it is best suited for species that occur in clustered distributions (where the problem can be

  16. Tensorial spacetime geometries carrying predictive, interpretable and quantizable matter dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Rivera Hernandez, Sergio

    2012-02-15

    Which tensor fields G on a smooth manifold M can serve as a spacetime structure? In the first part of this thesis, it is found that only a severely restricted class of tensor fields can provide classical spacetime geometries, namely those that can carry predictive, interpretable and quantizable matter dynamics. The obvious dependence of this characterization of admissible tensorial spacetime geometries on specific matter is not a weakness, but rather presents an insight: it was Maxwell theory that justified Einstein to promote Lorentzian manifolds to the status of a spacetime geometry. Any matter that does not mimick the structure of Maxwell theory, will force us to choose another geometry on which the matter dynamics of interest are predictive, interpretable and quantizable. These three physical conditions on matter impose three corresponding algebraic conditions on the totally symmetric contravariant coefficient tensor field P that determines the principal symbol of the matter field equations in terms of the geometric tensor G: the tensor field P must be hyperbolic, time-orientable and energy-distinguishing. Remarkably, these physically necessary conditions on the geometry are mathematically already sufficient to realize all kinematical constructions familiar from Lorentzian geometry, for precisely the same structural reasons. This we were able to show employing a subtle interplay of convex analysis, the theory of partial differential equations and real algebraic geometry. In the second part of this thesis, we then explore general properties of any hyperbolic, time-orientable and energy-distinguishing tensorial geometry. Physically most important are the construction of freely falling non-rotating laboratories, the appearance of admissible modified dispersion relations to particular observers, and the identification of a mechanism that explains why massive particles that are faster than some massless particles can radiate off energy until they are slower than all

  17. FPGA-Based Synthesis of High-Speed Hybrid Carry Select Adders

    Directory of Open Access Journals (Sweden)

    V. Kokilavani

    2015-01-01

    Full Text Available Carry select adder is a square-root time high-speed adder. In this paper, FPGA-based synthesis of conventional and hybrid carry select adders are described with a focus on high speed. Conventionally, carry select adders are realized using the following: (i full adders and 2 : 1 multiplexers, (ii full adders, binary to excess 1 code converters, and 2 : 1 multiplexers, and (iii sharing of common Boolean logic. On the other hand, hybrid carry select adders involve a combination of carry select and carry lookahead adders with/without the use of binary to excess 1 code converters. In this work, two new hybrid carry select adders are proposed involving the carry select and section-carry based carry lookahead subadders with/without binary to excess 1 converters. Seven different carry select adders were implemented in Verilog HDL and their performances were analyzed under two scenarios, dual-operand addition and multioperand addition, where individual operands are of sizes 32 and 64-bits. In the case of dual-operand additions, the hybrid carry select adder comprising the proposed carry select and section-carry based carry lookahead configurations is the fastest. With respect to multioperand additions, the hybrid carry select adder containing the carry select and conventional carry lookahead or section-carry based carry lookahead structures produce similar optimized performance.

  18. Functional muscle ischemia in Duchenne and Becker muscular dystrophy

    Directory of Open Access Journals (Sweden)

    Gail D Thomas

    2013-12-01

    Full Text Available Duchenne and Becker muscular dystrophy (DMD/BMD comprise a spectrum of devastating X-linked muscle wasting disease for which there is no treatment. DMD/BMD is caused by mutations in the gene encoding dystrophin, a cytoskeletal protein that stabilizes the muscle membrane and also targets other proteins to the sarcolemma. Among these is the muscle-specific isoform of neuronal nitric oxide synthase (nNOSµ which binds spectrin-like repeats within dystrophin’s rod domain and the adaptor protein α-syntrophin. Dystrophin deficiency causes loss of sarcolemmal nNOSµ and reduces paracrine signaling of muscle-derived nitric oxide (NO to the microvasculature, which renders the diseased muscle fibers susceptible to functional muscle ischemia during exercise. Repeated bouts of functional ischemia superimposed on muscle fibers already weakened by dystrophin deficiency result in use-dependent focal muscle injury. Genetic and pharmacologic strategies to boost nNOSµ-NO signaling in dystrophic muscle alleviate functional muscle ischemia and show promise as novel therapeutic interventions for the treatment of DMD/BMD.

  19. Correlates of weapon carrying in school among adolescents in three countries.

    Science.gov (United States)

    Stickley, Andrew; Koyanagi, Ai; Koposov, Roman; Blatný, Marek; Hrdlička, Michal; Schwab-Stone, Mary; Ruchkin, Vladislav

    2015-01-01

    To determine the factors associated with weapon carrying in school among Czech, Russian, and US adolescents. Logistic regression was used to analyze data drawn from the Social and Health Assessment (SAHA). Violent behavior (perpetration / victimization) was linked to adolescent weapon carrying in all countries. Substance use was associated with weapon carrying among boys in all countries. Greater parental warmth reduced the odds for weapon carrying among Czech and Russian adolescents. Associating with delinquent peers was important for weapon carrying only among US adolescents. Factors associated with weapon carrying in school vary among countries although violent behavior and substance use may be associated with weapon carrying across countries.

  20. 基于组间进位预测的快速进位加法器%Rapid Carry Adder Based on Carry Forecast Between Groups

    Institute of Scientific and Technical Information of China (English)

    丁宜栋; 刘昌明; 方湘艳

    2011-01-01

    为加快密码系统中大数加法的运算速度,提出并实现一种基于组间进位预测的快速进位加法器.将参与加法运算的大数进行分组,每个分组采用改进的超前进位技术以减少组内进位延时,组间通过进位预测完成不同进位状态下的加法运算,通过每个组产生的进位状态判断最终结果.性能分析表明,该进位加法器实现1024位大数加法运算的速度较快.%This paper presents and realizes a rapid carry adder based on carry forecast between groups to improve the speed of the large numbers adder in some cryptography systems. The large numbers is divided into many groups, the delay of carry-chain is reduced by carry lookahead in group. The group addition operation of different carry state is finished by carry forecast between groups. The addition sun of different carry forecast state is selected as the final result based on the carry state. Performance analysis shows that the computing speed of the carry adder is faster when realizing the 1 024 bit large numbers add operation.

  1. Dynamic Carrying Capacity Analysis of Double-Row Four-Point Contact Ball Slewing Bearing

    OpenAIRE

    Yunfeng Li; Di Jiang

    2015-01-01

    Carrying capacity is the most important performance index for slewing bearings. Maximizing the carrying capacity of slewing bearing is one pursuing goal for the bearing designer; this is usually realized by optimizing the design parameters. A method of analyzing the carrying capacity of double-row four-point contact ball slewing bearing by using dynamic carrying capacity surfaces was proposed in this paper. Based on the dynamic load carrying capacity surface of the slewing bearing, the effect...

  2. [Ecotourism carrying capacity of Hangzhou Xixi National Wetland Park in China].

    Science.gov (United States)

    Li, Rui; Rong, Liang

    2007-10-01

    In this paper, an integrated estimation on the ecotourism carrying capacity of Hangzhou Xixi National Wetland Park in China was made from the aspects of ecological carrying capacity, spatial carrying capacity, facility carrying capacity, management carrying capacity, and psychological carrying capacity. The results indicated that the tourism carrying capacity of the Park was 4 145 - 6 450 persons per day. The rational distance between man and bird was first adopted to determine the ecotourism carrying capacity of wetland, which provided an effective solution both to fully ensure bird safety and to appropriately develop wetland tourism. The estimation of psychological carrying capacity based on tourist satisfaction degree reflected more objectively the extent the tourist demands satisfied at the planning, construction and management of tour places. Such an integrated estimation method based on the distance between man and bird and the tourist satisfaction degree could be of practical and instructive significances in the planning and management of wetland parks.

  3. Weapon carrying and psychopathic-like features in a population-based sample of Finnish adolescents.

    Science.gov (United States)

    Saukkonen, Suvi; Laajasalo, Taina; Jokela, Markus; Kivivuori, Janne; Salmi, Venla; Aronen, Eeva T

    2016-02-01

    We investigated the prevalence of juvenile weapon carrying and psychosocial and personality-related risk factors for carrying different types of weapons in a nationally representative, population-based sample of Finnish adolescents. Specifically, we aimed to investigate psychopathic-like personality features as a risk factor for weapon carrying. The participants were 15-16-year-old adolescents from the Finnish self-report delinquency study (n = 4855). Four different groups were formed based on self-reported weapon carrying: no weapon carrying, carrying knife, gun or other weapon. The associations between psychosocial factors, psychopathic-like features and weapon carrying were examined with multinomial logistic regression analysis. 9% of the participants had carried a weapon in the past 12 months. Adolescents with a history of delinquency, victimization and antisocial friends were more likely to carry weapons in general; however, delinquency and victimization were most strongly related to gun carrying, while perceived peer delinquency (antisocial friends) was most strongly related to carrying a knife. Better academic performance was associated with a reduced likelihood of carrying a gun and knife, while feeling secure correlated with a reduced likelihood of gun carrying only. Psychopathic-like features were related to a higher likelihood of weapon carrying, even after adjusting for other risk factors. The findings of the study suggest that adolescents carrying a weapon have a large cluster of problems in their lives, which may vary based on the type of weapon carried. Furthermore, psychopathic-like features strongly relate to a higher risk of carrying a weapon.

  4. Psychophysical determination of load carrying capacity for a 1-h work period by Chinese males.

    Science.gov (United States)

    Wu, S P; Chen, C C

    2001-09-15

    This study used the psychophysical approach to examine the effects of container width, the presence or absence of container handles, and different load-carrying frequencies and distances on the maximum acceptable weight carried and the resulting response (heart rate and rating of perceived exertion) by well-conditioned males for a 1-h work period. After training, 12 male subjects performed a load-carrying task at knuckle height. Each subject performed 30 different carrying combinations. The conditions examined were container width, from 15.2 to 55.9 cm; carrying frequency, from 1 carry to 5 carries/min; and carrying distance from 1 to 6 m. The results were compared with prior studies and led to the following conclusions: (1) the use of container handles leads to the subjects carrying a significantly higher maximum acceptable weight than when containers do not have handles, which differs from the results of a previous study by Morrissey and Liou; (2) there were significant reductions in the maximum acceptable carrying weight with increases in container width, frequency and distance; (3) the presence or absence of container handles, different frequencies and load-carrying distances had significant effects on heart rate, although the effect of container width was not significant. In addition, the various frequencies and distances for load carrying had significant interaction effects on heart rate; (4) the effects of various frequencies and load-carrying distances on the rating of perceived exertion were statistically significant. The most stressed body parts were the wrists and arms.

  5. Sdelki carry trade ne roskosh, a sredstvo nakoplenija / Darius Gecevicius, Dmitri Fokin

    Index Scriptorium Estoniae

    Gecevicius, Darius

    2007-01-01

    Strateegia carry trade kasutamisest valuutaturgudel. Carry trade on tehing, mis enamasti toimub valuutaturul, kus investor laenab raha sisse valuutas, mille intressid on madalamad ning paigutab selle valuutas, kus intressid on kõrgemad. Diagramm

  6. Sdelki carry trade ne roskosh, a sredstvo nakoplenija / Darius Gecevicius, Dmitri Fokin

    Index Scriptorium Estoniae

    Gecevicius, Darius

    2007-01-01

    Strateegia carry trade kasutamisest valuutaturgudel. Carry trade on tehing, mis enamasti toimub valuutaturul, kus investor laenab raha sisse valuutas, mille intressid on madalamad ning paigutab selle valuutas, kus intressid on kõrgemad. Diagramm

  7. 46 CFR 180.72 - Personal flotation devices carried in addition to life jackets.

    Science.gov (United States)

    2010-10-01

    ... Jackets § 180.72 Personal flotation devices carried in addition to life jackets. (a) Equipment carried... jackets and must not be substituted for the approved life jackets required to be worn during drills and... 46 Shipping 7 2010-10-01 2010-10-01 false Personal flotation devices carried in addition to life...

  8. 36 CFR 1005.4 - Commercial passenger-carrying motor vehicles.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Commercial passenger-carrying motor vehicles. 1005.4 Section 1005.4 Parks, Forests, and Public Property PRESIDIO TRUST COMMERCIAL AND PRIVATE OPERATIONS § 1005.4 Commercial passenger-carrying motor vehicles. Passenger-carrying...

  9. 49 CFR 395.3 - Maximum driving time for property-carrying vehicles.

    Science.gov (United States)

    2010-10-01

    ... used by it to drive a property-carrying commercial motor vehicle, nor shall any such driver drive a property-carrying commercial motor vehicle: (1) More than 11 cumulative hours following 10 consecutive... commercial motor vehicle to drive, nor shall any driver drive a property-carrying commercial motor...

  10. Campaign for Right to Carry Concealed Guns on Campuses Gains Traction

    Science.gov (United States)

    Wiseman, Rachel

    2012-01-01

    Five years after the Virginia Tech massacre prompted a student in Texas to start a Facebook page in favor of the right to carry concealed weapons on college campuses, the resulting group, Students for Concealed Carry, has advanced its cause. In more than a dozen states, legislation has been introduced to allow the carrying of concealed firearms on…

  11. 46 CFR 150.160 - Carrying a cargo as an exception to the compatibility chart.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Carrying a cargo as an exception to the compatibility... BULK DANGEROUS CARGOES COMPATIBILITY OF CARGOES § 150.160 Carrying a cargo as an exception to the compatibility chart. The Operator of a vessel having on board a cargo carried as an exception under §...

  12. 25 CFR 161.204 - How are carrying capacities and stocking rates established?

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false How are carrying capacities and stocking rates... WATER NAVAJO PARTITIONED LANDS GRAZING PERMITS General Provisions § 161.204 How are carrying capacities..., review and adjust the carrying capacity of each range unit by determining the number of livestock,...

  13. Design of High-Speed Hybrid Carry Select Adders using VHDL

    OpenAIRE

    Mr. Vijay V. Gotmare; Dr. Pankaj Agarwal

    2016-01-01

    Carry select adder (CSA) is a square-root time high-speed adder. CSA is one of the fastest adders used in many data processing systems to perform fast arithmetic operations. In this project we propose to design hybrid carry select adders with a focus on high speed. CSA is a compromise between the longer delay Ripple carry adder (RCA) and the shorter delay Carry look-ahead adder (CLA). Conventionally carry select adders are realize using the full adders and 2:1 multiplexers. On the other hand ...

  14. The energetic costs of load-carrying and the evolution of bipedalism.

    Science.gov (United States)

    Watson, J C; Payne, R C; Chamberlain, A T; Jones, R K; Sellers, W I

    2008-05-01

    The evolution of habitual bipedalism is still a fundamental yet unsolved question for paleoanthropologists, and carrying is popular as an explanation for both the early adoption of upright walking and as a positive selection pressure once a terrestrial lifestyle had been adopted. However, to support or reject any hypothesis that suggests carrying efficiency was an important selective pressure, we need quantitative data on the costs of different forms of carrying behavior, especially infant-carrying since reduction in the grasping capabilities of the foot would have prevented infants from clinging on for long durations. In this study, we tested the hypothesis that the mode of load carriage influences the energetic cost of locomotion. Oxygen consumption was measured in seven female participants walking at a constant speed while carrying four different 10-kg loads (a weighted vest, 5-kg dumbbells carried in each hand, a mannequin infant carried on one hip, and a 10-kg dumbbell carried in a single hand). Oxygen consumption was also measured during unloaded standing and unloaded walking. The results show that the weighted vest requires the least amount of energy of the four types of carrying and that, for this condition, humans are as efficient as mammals in general. The balanced load was carried with approximately the predicted energy cost. However, the asymmetrical conditions were considerably less efficient, indicating that, unless infant-carrying was the adaptive response to a strong environmental selection pressure, this behavior is unlikely to have been the precursor to the evolution of bipedalism.

  15. Dynamic Carrying Capacity Analysis of Double-Row Four-Point Contact Ball Slewing Bearing

    Directory of Open Access Journals (Sweden)

    Yunfeng Li

    2015-01-01

    Full Text Available Carrying capacity is the most important performance index for slewing bearings. Maximizing the carrying capacity of slewing bearing is one pursuing goal for the bearing designer; this is usually realized by optimizing the design parameters. A method of analyzing the carrying capacity of double-row four-point contact ball slewing bearing by using dynamic carrying capacity surfaces was proposed in this paper. Based on the dynamic load carrying capacity surface of the slewing bearing, the effect of changes of the bearing design parameters, such as axial clearance, raceway groove radius coefficient, and contact angle, on the dynamic carrying capacity of the slewing bearing was researched; the trend and the degree of the effect of the micro changes of the bearing design parameters on the dynamic load carrying capacity of the bearing were discussed, and the results provide the basis for optimizing the design parameter of this type of slewing bearing.

  16. A model to estimate aquaculture carrying capacity in three areas of the Philippines

    Directory of Open Access Journals (Sweden)

    Tarzan Legović

    2008-12-01

    Full Text Available A model was developed to estimate the production carrying capacity of water bodies based on nutrient inputs from aquaculture and other sources, flushing rates, and the risk of algal blooms for three different areas of the Philippines – Bolinao (marine site, Dagupan (brackishwater site and Taal Lake (freshwater site. The results suggest that aquaculture production in the Taal Lake was greater than the sustainable carrying capacity. Aquaculture structures in Bolinao were close to carrying capacity during average tidal exchange but greater than the carrying capacity during low tidal exchange and no winds.Aquaculture production in the Dagupan estuary has not overcome its carrying capacity even during low flow. However, during very low flow and no tidal flushing, carrying capacity has been overcome.

  17. Design & Analysis of Low Power, Area-Efficient Carry Select Adder

    Directory of Open Access Journals (Sweden)

    Shuchi Verma

    2014-03-01

    Full Text Available This paper deals with the design & analysis of Carry Select Adder (CSLA & Carry lookahead adder (CLA. Adders are designed using 0.18µm CMOS process technology & simulated with Modelsim6.3f. The adder designs, Regular CSLA, modified CSLA using BEC, modified CSLA without using multiplexer, modified CSLA using D-Latch & Carry lookahead adders in 4-bit, 16-bit, 32-bit, are compared with the simulated results on the basis area.

  18. Synchronisation of fertility with carrying capacity; an investigation using classical and agent based modeling

    Directory of Open Access Journals (Sweden)

    Hugh Crozier Murrell

    2013-05-01

    Full Text Available A generalized Verhulst model subject to seasonal change in both fertility rate and carrying capacity is outlined. Numerical solutions to the Verhulst equations are employed to obtain optimal fertility rate phase shift with respect to carrying capacity. Possible natural selection for a preferred season of conception is investigated using agent based simulations. Both experiments indicate that synchronization of fertility rate to environment carrying capacity is beneficial to species survival.

  19. Research on Carrying Capacity of Water Resource Based on Multi-reservoir Regulation

    OpenAIRE

    Liu Renyuan; Li Lei; Li Hua; Huang Qiang

    2013-01-01

    Analyze the level of Carrying Capacity of Water Resources; evaluate the condition of e region social, economic and ecological environment comprehensive and sustainable development. This study is based on analyzing the meaning of water resources carrying capacity, take water resources of the Midwest of Shenzhen City an example., by the way of the “Coordinated and sustainable development” theory, found the model of the Carrying Capacity of Water Resources, Calculated and analyzed. Water efficie...

  20. Effects of a Fluctuating Carrying Capacity on the Generalized Malthus-Verhulst Model

    OpenAIRE

    Héctor Calisto; Chandía, Kristopher J.; Mauro Bologna

    2014-01-01

    We consider a generalized Malthus-Verhulst model with a fluctuating carrying capacity and we study its effects on population growth. The carrying capacity fluctuations are described by a Poissonian process with an exponential correlation function. We will find an analytical expression for the average of a number of individuals and show that even in presence of a fluctuating carrying capacity the average tends asymptotically to a constant quantity.

  1. Estimating Recreational Carrying Capacity of Gheisary Protected Area in Chaharmahal & Bakhtiari Province

    OpenAIRE

    Sheikh, A; Jafari, A.; Setoodeh, A

    2014-01-01

    It is now certain that to ensure a long term sustainable development, protected areas, the favored destinations for ecotourism, required a well written planning management. Each destination however has limited capability which is called “carrying capacity” to invite visitors. Physical, real (ecological) and effective (manageable) are different kinds of carrying capacity. The current study presented these varieties of carrying capacities for Gheisary protected area, stretched over 10,000 ha in...

  2. ANALYSIS OF ABILITY FOR IGNITING METHANE OF WATER-CARRYING EXPLOSIVES

    Institute of Scientific and Technical Information of China (English)

    陆守香

    1997-01-01

    In this paper, the ability to ignite methane of water-carrying explosives is discussed in detail. The difference of the safety towards methane of water- carrying explosives and ammonite is analysed comparatively. It is shown that the process of detonation reaction, the duration of flame by combustible residues of detonation and the fineness of cooling salt in detonation products are important factors of the safety of explosive towards methane. Water-carrying explosives are safer than ammonite used in coal mines.

  3. Effects of a Fluctuating Carrying Capacity on the Generalized Malthus-Verhulst Model

    Directory of Open Access Journals (Sweden)

    Héctor Calisto

    2014-01-01

    Full Text Available We consider a generalized Malthus-Verhulst model with a fluctuating carrying capacity and we study its effects on population growth. The carrying capacity fluctuations are described by a Poissonian process with an exponential correlation function. We will find an analytical expression for the average of a number of individuals and show that even in presence of a fluctuating carrying capacity the average tends asymptotically to a constant quantity.

  4. The Association Between Weapon Carrying and Health Risk Behaviors Among Adolescent Students in Bangkok, Thailand.

    Science.gov (United States)

    Saiphoklang, On-Anong; Wongboonsin, Kua; Wongboonsin, Patcharawalai; Perngparn, Usaneya; Cottler, Linda B

    2015-07-30

    Carrying weapons is a significant social and public health problem worldwide, especially among adolescents. The present study examined the association between weapon carrying and related risk behaviors among Thai adolescents. A cross-sectional study of 2,588 high school and vocational school students aged 11 to 19 years from 26 schools in Bangkok, Thailand, was conducted in 2014. This study found that 7.8% of youth reported having carried a weapon in the past 12 months. The high prevalence of weapon carrying was reported by male students, and males were more likely to have reported carrying a weapon than females. The association between weapon carrying and the health risk behaviors like drinking, smoking, any drug use, and physical fighting were significant with higher odds of weapon carrying in all models. Among males, weapon carrying was related to drinking and smoking, any drug use, physical fighting, and school type. Among females, suicidal thoughts were significantly related along with drinking and smoking, any drug use, and physical fighting. Having a mother who used substances was significant only among females. These data could be used for further interventions about weapon carrying to reduce violence. © The Author(s) 2015.

  5. Review on the Evaluation System of Public Safety Carrying Capacity about Small Town Community

    Institute of Scientific and Technical Information of China (English)

    Ming; SUN; Tianyu; ZHU

    2014-01-01

    Recently,small town community public safety problem has been increasingly highlighted,but its research is short on public safety carrying capacity. Through the investigation and study of community public safety carrying capacity,this paper analyzes the problem of community public safety in our country,to construct index evaluation system of public safety carrying capacity in small town community. DEA method is used to evaluate public safety carrying capacity in small town community,to provide scientific basis for the design of support and standardization theory about small town community in public safety planning.

  6. An Area-Efficient Carry Select Adder Design by using 180 nm Technology

    OpenAIRE

    Garish Kumar Wadhwa; Amit Grover; Neeti Grover; GurpreetSingh

    2013-01-01

    In this paper, we proposed an area-efficient carry select adder by sharing the common Boolean logic term. After logic simplification and sharing partial circuit, we only need one XOR gate and one inverter gate in each summation operation as well as one AND gate and one inverter gate in each carry-out operation. Through the multiplexer, we can select the correct output result according to the logic state of carry-in signal. In this way, the transistor count in a 32-bit carry select adder can b...

  7. An Area-Efficient Carry Select Adder Design by using 180 nm Technology

    Directory of Open Access Journals (Sweden)

    Garish Kumar Wadhwa

    2013-02-01

    Full Text Available In this paper, we proposed an area-efficient carry select adder by sharing the common Boolean logic term. After logic simplification and sharing partial circuit, we only need one XOR gate and one inverter gate in each summation operation as well as one AND gate and one inverter gate in each carry-out operation. Through the multiplexer, we can select the correct output result according to the logic state of carry-in signal. In this way, the transistor count in a 32-bit carry select adder can be greatly reduced from 1947 to 960.

  8. Specifying the Role of Exposure to Violence and Violent Behavior on Initiation of Gun Carrying: A Longitudinal Test of Three Models of Youth Gun Carrying

    Science.gov (United States)

    Spano, Richard; Pridemore, William Alex; Bolland, John

    2012-01-01

    Two waves of longitudinal data from 1,049 African American youth living in extreme poverty are used to examine the impact of exposure to violence (Time 1) and violent behavior (Time 1) on first time gun carrying (Time 2). Multivariate logistic regression results indicate that (a) violent behavior (Time 1) increased the likelihood of initiation of…

  9. Specifying the Role of Exposure to Violence and Violent Behavior on Initiation of Gun Carrying: A Longitudinal Test of Three Models of Youth Gun Carrying

    Science.gov (United States)

    Spano, Richard; Pridemore, William Alex; Bolland, John

    2012-01-01

    Two waves of longitudinal data from 1,049 African American youth living in extreme poverty are used to examine the impact of exposure to violence (Time 1) and violent behavior (Time 1) on first time gun carrying (Time 2). Multivariate logistic regression results indicate that (a) violent behavior (Time 1) increased the likelihood of initiation of…

  10. 46 CFR 117.72 - Personal flotation devices carried in addition to life jackets.

    Science.gov (United States)

    2010-10-01

    ... PASSENGERS LIFESAVING EQUIPMENT AND ARRANGEMENTS Ring Life Buoys and Life Jackets § 117.72 Personal flotation devices carried in addition to life jackets. (a) Equipment carried under this section is not acceptable in lieu of any portion of the required number of approved life jackets and must not be substituted for the...

  11. Risk-premia, Carry-trade Dynamics, and Economic Value of Currency Speculation

    DEFF Research Database (Denmark)

    Wagner, Christian

    2012-01-01

    -cited ‘forward bias puzzle’ originates from the omission of the risk-premium in standard UIP tests. Consistent with its popularity among market professionals, the carry-trade strategy can be rationalized as it systematically collects risk-premia, however, the economic value generated by bilateral carry-trades...

  12. Form and Function of Carrying Tools in Traditional and Contemporary Culture

    Directory of Open Access Journals (Sweden)

    Hendriana Werdhaningsih

    2009-07-01

    Full Text Available The Javanese Traditional Carrying Tools are categorized into two kinds based used of materials: anyaman made of woven bamboo or rattan and the other was made of cloth. The Traditional Javanese Carrying Tools that were made of bamboo/ rattan is actually a kind of container with different shapes. The types of goods that were brought was related to the shapes of the containers, demonstrate the type of interactions between the carrier and the goods. The Traditional Carrying Tools made of cloth have the flexibility in terms of being the containers of the carried goods. Selendang is used to carry goods, including the bamboo/ rattan containers and to carry babies. It can be used rumpled and straightened depends on the technical needs of carrying tools preferred by the user. In contemporary culture, the form and design of carrying tools more less construct by fashion and trends besides those practical used. Some product are being 'classic', some other are being 'fad'. Both products, traditional and contemporary have their own style, uniqueness, and own context. Analyzing both in design point of view is important as evaluation process, to finding new problems that will be starting point to create new products. Indonesia is well known for its abundant richness in traditions, which include the culture, the art, and the traditional products. Such treasure of traditions can inspire the development and invention of various kinds of artwork and current products.

  13. Complete genome sequences of Incl1 Plasmids carrying extended-spectrum B-Lactamase genes

    NARCIS (Netherlands)

    Brouwer, M.S.M.; Bossers, A.; Harders, F.; Essen-Zandbergen, van A.; Mevius, D.J.; Smith, H.E.

    2014-01-01

    Extended spectrum beta-lactamases (ESBLs) confer resistance to clinically relevant antibiotics. Often, the resistance genes are carried by conjugative plasmids which are responsible for dissemination. Five IncI1 plasmids carrying ESBLs from commensal and clinical Escherichia coli isolates were compl

  14. Gun Cultures or Honor Cultures? Explaining Regional and Race Differences in Weapon Carrying

    Science.gov (United States)

    Felson, Richard B.; Pare, Paul-Philippe

    2010-01-01

    We use the National Violence against Women (and Men) Survey to examine the effects of region and race on the tendency to carry weapons for protection. We find that Southern and Western whites are much more likely than Northern whites to carry guns for self-protection, controlling for their risk of victimization. The difference between Southern and…

  15. Predictors of Weapon Carrying in Youth Attending Drop-in Centers

    Science.gov (United States)

    Blumberg, Elaine J.; Liles, Sandy; Kelley, Norma J.; Hovell, Melbourne F.; Bousman, Chad A.; Shillington, Audrey M.; Ji, Ming; Clapp, John

    2009-01-01

    Objective: To test and compare 2 predictive models of weapon carrying in youth (n=308) recruited from 4 drop-in centers in San Diego and Imperial counties. Methods: Both models were based on the Behavioral Ecological Model (BEM). Results: The first and second models significantly explained 39% and 53% of the variance in weapon carrying,…

  16. Carrying a Weapon to School and Perceptions of Social Support in an Urban Middle School.

    Science.gov (United States)

    Malecki, Christine Kerres; Demaray, Michelle Kilpatrick

    2003-01-01

    This study of perceived social support and weapon possession surveyed 461 students in an urban middle school. Students who reported carrying weapons to school reported less overall or total perceived social support (from peers, parents, teachers, classmates, and school) than did their peers who did not carry weapons. Perceived social support was a…

  17. Youths Carrying a Weapon or Using a Weapon in a Fight: What Makes the Difference?

    Science.gov (United States)

    Thurnherr, Judit; Michaud, Pierre-Andre; Berchtold, Andre; Akre, Christina; Suris, Joan-Carles

    2009-01-01

    The objective of this study was to characterize weapon-carrying adolescents and to assess whether weapon carriers differ from weapon users. Data were drawn from a cross-sectional school-based survey of 7548 adolescents aged 16-20 years in Switzerland. Youths carrying a weapon were compared with those who do not. Subsequently, weapon carriers were…

  18. Effect of Gun Carrying on Perceptions of Risk Among Adolescent Offenders.

    Science.gov (United States)

    Loughran, Thomas A; Reid, Joan A; Collins, Megan Eileen; Mulvey, Edward P

    2016-02-01

    We observed how perceptions of risks, costs, crime rewards, and violence exposure change as individual gun-carrying behavior changes among high-risk adolescents. We analyzed a longitudinal study (2000-2010) of serious juvenile offenders in Maricopa County, Arizona, or Philadelphia County, Pennsylvania, assessing within-person changes in risk and reward perceptions, and violence exposure as individuals initiated or ceased gun carrying. Despite being associated with heightened exposure to violence, gun carrying was linked to lower perceptions of risks and costs and higher perceived rewards of offending. Gun carrying was not time-stable, as certain individuals both started and stopped carrying during the study. Within-person changes in carrying guns were associated with shifting perceptions of risks, costs, and rewards of crime, and changes in exposure to violence in expected directions. Gun carrying reduces perceptions of risks associated with offending while increasing actual risk of violence exposure. This suggests that there is an important disconnect between perceptions and objective levels of safety among high-risk youths. Gun-carrying decisions may not only be influenced by factors of protection and self-defense, but also by perceptions of risks and reward associated with engaging in crime more generally.

  19. 48 CFR 19.707 - The Small Business Administration's role in carrying out the program.

    Science.gov (United States)

    2010-10-01

    ... Subcontracting Program 19.707 The Small Business Administration's role in carrying out the program. (a) Under the... 48 Federal Acquisition Regulations System 1 2010-10-01 2010-10-01 false The Small Business Administration's role in carrying out the program. 19.707 Section 19.707 Federal Acquisition Regulations...

  20. Gun Cultures or Honor Cultures? Explaining Regional and Race Differences in Weapon Carrying

    Science.gov (United States)

    Felson, Richard B.; Pare, Paul-Philippe

    2010-01-01

    We use the National Violence against Women (and Men) Survey to examine the effects of region and race on the tendency to carry weapons for protection. We find that Southern and Western whites are much more likely than Northern whites to carry guns for self-protection, controlling for their risk of victimization. The difference between Southern and…

  1. The application of a phosphorus mass balance model for estimating the carrying capacity of Lake Kariba

    OpenAIRE

    MHLANGA, Lindah; MHLANGA, Wilson; MWERA, Paul

    2013-01-01

    The aim of this study was to use an empirical mass balance equation to estimate the carrying capacity of Lake Kariba, where cage culture for Nile tilapia Oreochromis niloticus has been practiced since 1996. The carrying capacity for the lake was estimated at 33.2 × 103 t per year using the Dillon-Rigler phosphorus budget model.

  2. CERTAINTY EQUIVALENCE FOR DETERMINATION OF OPTIMAL FERTILIZER APPLICATION RATES WITH CARRY-OVER

    OpenAIRE

    Taylor, C. Robert

    1983-01-01

    This note demonstrates that a certain class of stochastic problems for determination of optimal fertilizer application rates in the presence of fertilizer carry-over can be simplified to static, certainly equivalent problems. Conditions required for certainty equivalence to hold are: (1) fertilizer carry-over is agronomically equivalent to applied fertilizer; and (2) some addition of fertilizer is optimal in every decision period.

  3. Toddlers actively reorganize their whole body coordination to maintain walking stability while carrying an object.

    Science.gov (United States)

    Hsu, Wen-Hao; Miranda, Daniel L; Chistolini, Trevor L; Goldfield, Eugene C

    2016-10-01

    Balanced walking involves freely swinging the limbs like pendula. However, children immediately begin to carry objects as soon as they can walk. One possibility for this early skill development is that whole body coordination during walking may be re-organized into loosely coupled collections of body parts, allowing children to use their arms to perform one function, while the legs perform another. Therefore, this study examines: 1) how carrying an object affects the coordination of the arms and legs during walking, and 2) if carrying an object influences stride length and width. Ten healthy toddlers with 3-12 months of walking experience were recruited to walk barefoot while carrying or not carrying a small toy. Stride length, width, speed, and continuous relative phase (CRP) of the hips and of the shoulders were compared between carrying conditions. While both arms and legs demonstrated destabilization and stabilization throughout the gait cycle, the arms showed a reduction in intra-subject coordination variability in response to carrying an object. Carrying an object may modify the function of the arms from swinging for balance to maintaining hold of an object. The observed period-dependent changes of the inter-limb coordination of the hips and of the shoulders also support this interpretation. Overall, these findings support the view that whole-body coordination patterns may become partitioned in particular ways as a function of task requirements.

  4. Phenomenological approach to describe logistic growth and carrying capacity-dependent growth processes

    Indian Academy of Sciences (India)

    DIBYENDU BISWAS; SWARUP PORIA; SANKAR NARAYAN PATRA

    2016-11-01

    In this communication, different classes of phenomenological universalities of carrying capacity dependent growth processes have been proposed. The logistic as well as carrying capacity-dependent West-type allometry-based biological growths can be explained in this proposed framework. It is shown that logistic and carrying capacity-dependent West-type growths are phenomenologically identical in nature. However, there is a difference between them in terms of coefficients involved in the phenomenological descriptions. Involuted Gompertz function, used to describe biological growth processes undergoing atrophy or a demographic and economic system undergoing involution or regression, can be addressed in this proposed environment-dependent description. It is also found phenomenologically that the energy intake of an organism depends on carrying capacity whereas metabolic cost does not depend on carrying capacity. In addition, some other phenomenologicaldescriptions have been examined in this proposed framework and graphical representations of variation of different parameters involved in the description are executed.

  5. A Feminist Reading of Sister Carrie: Carrie——the Aspiring Cinderella

    Institute of Scientific and Technical Information of China (English)

    殷娟

    2008-01-01

    Like Cinderella in the fairy tale Cinderella, Carrie is beautiful, innocent and poor. But unlike the traditional Cinderella who mindlessly waits around for a handsome prince to come to her rescue, Carrie, the aspiring Cinderella, resolvedly leaves the men she ever depended on and eventually becomes a successful actress. Sister Carrie, the most impressive heroine of Theodore Dreiser, is a contradictory character. Some accuse her of realizing her ambition with immoral means. However, some others take sympathy with her courage, determination, and aspiration. By analyzing the most distinguished quality of Sister Carrie - her aspiration, this essay is to illustrate from the perspective of feminism that women are not limited by their sex. Ignoring her own sex as well as the conventional attitude toward women in a male-dominated society, sister Carrie, full of aspiration, takes an active pan in creating and determining her own lives and her own futures..

  6. Research on Carrying Capacity of Water Resource Based on Multi-reservoir Regulation

    Directory of Open Access Journals (Sweden)

    Liu Renyuan

    2013-12-01

    Full Text Available Analyze the level of Carrying Capacity of Water Resources; evaluate the condition of e region social, economic and ecological environment comprehensive and sustainable development. This study is based on analyzing the meaning of water resources carrying capacity, take water resources of the Midwest of Shenzhen City an example., by the way of the “Coordinated and sustainable development” theory, found the model of the Carrying Capacity of Water Resources, Calculated and analyzed. Water efficiency coefficients are different in different states, the Carrying Capacity of Water Resources is different too. the water resources in the Midwest of Shenzhen City can only meet now this stage (2010 Urban water demand, meanwhile people of over loading in there are also constantly increasing, In the future, Water requirement of Shenzhen Midwest cities must increase Dongjiang water and west diversion works of Shen Zhen, in order to enhance Carrying Capacity of Water Resources of the area.

  7. Risk Analysis on Groundwater Resources Carrying Capacity Based on Blind Number Theory

    Institute of Scientific and Technical Information of China (English)

    ZHANG Ji; YU Sujun

    2007-01-01

    Blind numbers of evaluation indices about groundwater resources carrying capacity are defined from the concomitancy of randomness, fuzziness, grey property and unascertainment of groundwater system.Based on fuzzy theory, a comprehensive evaluation model on groundwaterresources carrying capacity is constructed with blind information. Then arisk assessment model of surcharge about groundwater resources carryingcapacity is established on blind reliability theory. The probable value"*"'matrix of fuzzy membership degree about carrying capacity corresponding to each judgment level can be obtained with the aid of blind algorithm as well as the subjective reliability "×" matrix. And then a graph of"groundwater carrying capacity v.s. accumulative reliability" can be gained.Based on the graph, fuzzy membership degree of groundwater resourcescarrying capacity to each judgment level under different risk probabilitycan be got. Thus, a comparatively reasonable judgment to groundwaterresources carrying capacity might be obtained, with comprehensive analysis to the state of society, economy technology and ecology.

  8. Comparative analysis of environmental carrying capacity of the Bohai Sea Rim area in China.

    Science.gov (United States)

    Lin, Lu; Liu, Yi; Chen, Jining; Zhang, Tianzhu; Zeng, Siyu

    2011-11-01

    Environmental carrying capacity is an essential metric for measuring regional sustainability. Although the term "carrying capacity" has been applied for over a century, the concept definition, quantitative methods and comprehensive evaluation remain arguable. This study analyzed the carrying capacity of four environmental elements, including water resources, air, surface water and offshore sea, and integrated them into a comprehensive index to represent overall regional profiles of resources and environment. The method was then applied to thirteen municipalities in the Bohai Sea Rim area, one of the most rapidly developing regions in transition China. The results show that the comprehensive environmental carrying capacity of the municipalities in the south sub-region were largest in 2007, while that of the west municipalities were lowest. The regional economic development exceeded the overall environmental carrying capacity by 36% and the west sub-region area deserves overwhelming attention for future industrial allocation.

  9. Object carrying as socio-sexual display in an aquatic mammal.

    Science.gov (United States)

    Martin, A R; da Silva, V M F; Rothery, P

    2008-06-23

    Amazon river dolphins or botos (Inia geoffrensis Blainville) were observed carrying objects in 221 social groups over a 3-year study period. Sticks, branches and clumps of grass were taken from the water surface and often repeatedly thrashed or thrown. Lumps of hard clay were collected from the river bed and held in the mouth while the carrier rose slowly above the surface and submerged again. Carriers were predominantly adult males and less often subadult males. Adult females and young dolphins rarely carried objects. Groups of dolphins in which object carrying occurred were differentially large and comprised a greater proportion of adult males and adult females. Aggression, mostly between adult males, was significantly associated with object carrying. The behaviour occurred year-round, with peaks in March and July. A plausible explanation of the results is that object carrying by adult males is aimed at females and is stimulated by the number of females in the group, while aggression is targeted at adult males and is stimulated by object carrying in the group. We infer that object carrying in this sexually dimorphic species is socio-sexual display. It is either of ancient origin or has evolved independently in several geographically isolated populations.

  10. Ecological Carrying Capacity in Beijing Mountainous Areas Taking Fangshan District as an Example

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    [Objective] The aim was to investigate ecological carrying capacity of Fangshan District in Beijing. [Method] Ecological footprint and carrying capacity of Fangshan District in 2007 and 2008 were analyzed with ecological footprint model and the difference between ecological footprint and carrying capacity there was con- cluded. Furthermore, an assessment was made on relation between Fangshan de- velopment with local ecological carrying capacity to evaluate sustainable development of Fangshan District. [Result] Fangshan District is of lightly ecological deficit; ecologi- cal footprint per capita in 2007-2008 was deficit and tended to expand. Two meth- ods are available to reduce ecological :deficit, including reduction of ecological foot- print and increase of carrying capacity. In general, supply and demand of carrying capacity in Fangshan are better, though with some problems. It is necessary to fur- ther develop related industries to final eliminate ecological deficit in order to achieve ecological conservation-orientation and sustainable development of Fangshan District. [Conclusion] The research provides theoretical references for overall evaluation on eco- logical carrying capacity of Fangshan District.

  11. Physical and emotional health problems experienced by youth engaged in physical fighting and weapon carrying.

    Directory of Open Access Journals (Sweden)

    Sophie D Walsh

    Full Text Available Then aims of the current study were 1 to provide cross-national estimates of the prevalence of physical fighting and weapon carrying among adolescents aged 11-15 years; (2 To examine the possible effects of physical fighting and weapon carrying on the occurrence of physical (medically treated injuries and emotional health outcomes (multiple health complaints among adolescents within the theoretical framework of Problem Behaviour Theory. 20,125 adolescents aged 11-15 in five countries (Belgium, Israel, USA, Canada, FYR Macedonia were surveyed via the 2006 Health Behaviour in School Aged Children survey. Prevalence was calculated for physical fighting and weapon carrying along with physical and emotional measures that potentially result from violence. Regression analyses were used to quantify associations between violence/weapon carrying and the potential health consequences within each country. Large variations in fighting and weapon carrying were observed across countries. Boys reported more frequent episodes of fighting/weapon carrying and medically attended injuries in every country, while girls reported more emotional symptoms. Although there were some notable variations in findings between different participating countries, increased weapon carrying and physical fighting were both independently and consistently associated with more frequent reports of the potential health outcomes. Adolescents engaging in fighting and weapon carrying are also at risk for physical and emotional health outcomes. Involvement in fighting and weapon carrying can be seen as part of a constellation of risk behaviours with obvious health implications. Our findings also highlight the importance of the cultural context when examining the nature of violent behaviour for adolescents.

  12. Physical and emotional health problems experienced by youth engaged in physical fighting and weapon carrying.

    Science.gov (United States)

    Walsh, Sophie D; Molcho, Michal; Craig, Wendy; Harel-Fisch, Yossi; Huynh, Quynh; Kukaswadia, Atif; Aasvee, Katrin; Várnai, Dora; Ottova, Veronika; Ravens-Sieberer, Ulrike; Pickett, William

    2013-01-01

    Then aims of the current study were 1) to provide cross-national estimates of the prevalence of physical fighting and weapon carrying among adolescents aged 11-15 years; (2) To examine the possible effects of physical fighting and weapon carrying on the occurrence of physical (medically treated injuries) and emotional health outcomes (multiple health complaints) among adolescents within the theoretical framework of Problem Behaviour Theory. 20,125 adolescents aged 11-15 in five countries (Belgium, Israel, USA, Canada, FYR Macedonia) were surveyed via the 2006 Health Behaviour in School Aged Children survey. Prevalence was calculated for physical fighting and weapon carrying along with physical and emotional measures that potentially result from violence. Regression analyses were used to quantify associations between violence/weapon carrying and the potential health consequences within each country. Large variations in fighting and weapon carrying were observed across countries. Boys reported more frequent episodes of fighting/weapon carrying and medically attended injuries in every country, while girls reported more emotional symptoms. Although there were some notable variations in findings between different participating countries, increased weapon carrying and physical fighting were both independently and consistently associated with more frequent reports of the potential health outcomes. Adolescents engaging in fighting and weapon carrying are also at risk for physical and emotional health outcomes. Involvement in fighting and weapon carrying can be seen as part of a constellation of risk behaviours with obvious health implications. Our findings also highlight the importance of the cultural context when examining the nature of violent behaviour for adolescents.

  13. State Gun Law Environment and Youth Gun Carrying in the United States.

    Science.gov (United States)

    Xuan, Ziming; Hemenway, David

    2015-11-01

    Gun violence and injuries pose a substantial threat to children and youth in the United States. Existing evidence points to the need for interventions and policies for keeping guns out of the hands of children and youth. (1) To examine the association between state gun law environment and youth gun carrying in the United States, and (2) to determine whether adult gun ownership mediates this association. This was a repeated cross-sectional observational study design with 3 years of data on youth gun carrying from US states. The Youth Risk Behavior Survey comprises data of representative samples of students in grades 9 to 12 from biennial years of 2007, 2009, and 2011. We hypothesized that states with more restrictive gun laws have lower rates of youth gun carrying, and this association is mediated by adult gun ownership. State gun law environment as measured by state gun law score. Youth gun carrying was defined as having carried a gun on at least 1 day during the 30 days before the survey. In the fully adjusted model, a 10-point increase in the state gun law score, which represented a more restrictive gun law environment, was associated with a 9% decrease in the odds of youth gun carrying (adjusted odds ratio [AOR], 0.91 [95% CI, 0.86-0.96]). Adult gun ownership mediated the association between state gun law score and youth gun carrying (AOR, 0.94 [ 95% CI, 0.86-1.01], with 29% attenuation of the regression coefficient from -0.09 to -0.07 based on bootstrap resampling). More restrictive overall gun control policies are associated with a reduced likelihood of youth gun carrying. These findings are relevant to gun policy debates about the critical importance of strengthening overall gun law environment to prevent youth gun carrying.

  14. Impact of Flexibility Options on Grid Economic Carrying Capacity of Solar and Wind: Three Case Studies

    Energy Technology Data Exchange (ETDEWEB)

    Denholm, Paul [National Renewable Energy Lab. (NREL), Golden, CO (United States); Novacheck, Joshua [National Renewable Energy Lab. (NREL), Golden, CO (United States); Jorgenson, Jennie [National Renewable Energy Lab. (NREL), Golden, CO (United States); O' Connell, Matthew [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2016-12-01

    In this study, we attempt to quantify the benefits of various options of grid flexibility by measuring their impact on two measures: economic carrying capacity and system costs. Flexibility can increase economic carrying capacity and reduce overall system costs. In some cases, options that provide a limited increase in economic carrying capacity can provide significant operational savings, thus demonstrating the need to evaluate flexibility options using multiple metrics. The value of flexibility options varies regionally due to different generation mixes and types of renewables. The more rapid decline in PV value compared to wind makes PV more dependent on adding flexibility options, including transmission and energy storage.

  15. Glove powder's carrying capacity for latex protein: analysis using the ASTM ELISA test.

    Science.gov (United States)

    Beezhold, D; Horton, K; Hickey, V; Daddona, J; Kostyal, D

    2003-01-01

    Glove donning powders carry latex proteins and disperse them into the workplace environment. We have used the ASTM D6499 ELISA to quantify the amount of latex antigen bound to and carried by glove powders. We could differentiate between a small amount of protein actually bound to the powders and a larger amount carried by the powder. Enhanced binding of a major allergen, Hev b 5, to the starch powders was demonstrated by Western blot. The D6499 ELISA is able to measure total latex antigen, soluble and powder bound, simultaneously without the need to centrifuge the samples.

  16. Quantifying spatially derived carrying capacity occupation: Framework for characterisation modelling and application to terrestrial acidification

    DEFF Research Database (Denmark)

    Bjørn, Anders; Margni, M.; Bulle, C.

    *year. This metric resembles that of the ecological footprint method and may be compared to the availability of land or water. The framework was applied to the terrestrial acidification impact category. The geochemical steady-state model PROFILE was used to quantify carrying capacities as deposition levels......The popularity of the ecological footprint method and the planetary boundaries concept shows an increasing interest among decision makers in comparing environmental impacts to carrying capacities of natural systems. Recently carrying capacity-based normalisation references were developed for impact...

  17. Carrying Capacity Model Applied to Coastal Ecotourism of Baluran National Park, Indonesia

    Science.gov (United States)

    Armono, H. D.; Rosyid, D. M.; Nuzula, N. I.

    2017-07-01

    The resources of Baluran National Park have been used for marine and coastal ecotourism. The increasing number of visitors has led to the increasing of tourists and its related activities. This condition will cause the degradation of resources and the welfare of local communities. This research aims to determine the sustainability of coastal ecotourism management by calculating the effective number of tourists who can be accepted. The study uses the concept of tourism carrying capacity, consists the ecological environment, economic, social and physical carrying capacity. The results of the combined carrying capacity analysis in Baluran National Park ecotourism shows that the number of 3.288 people per day (151.248 tourists per year) is the maximum number of accepted tourists. The current number of tourist arrivals is only 241 people per day (87.990 tourists per year) which is far below the carrying capacity.

  18. Geometry of carrying simplices of 3-species competitive Lotka-Volterra systems

    Science.gov (United States)

    Baigent, Stephen

    2013-04-01

    We investigate the existence, uniqueness and Gaussian curvature of the invariant carrying simplices of 3 species autonomous totally competitive Lotka-Volterra systems. Explicit examples are given where the carrying simplex is convex or concave, but also where the curvature is not single-signed. Our method monitors the curvature of an evolving surface that converges uniformly to the carrying simplex, and generally relies on establishing that the Gaussian image of the evolving surface is confined to an invariant cone. We also discuss the relationship between the curvature of the carrying simplex near an interior fixed point and its Split Lyapunov stability. Finally we comment on extensions to general Lotka-Volterra systems that are not competitive.

  19. 76 FR 16239 - Moratorium on New Exemptions for Passenger Carrying Operations Conducted for Compensation and...

    Science.gov (United States)

    2011-03-23

    ... holder to operate a large, crew-served, piston-powered, multiengine, World War II (WWII) bomber carrying... experience flight in these aircraft. A 2004 policy explicitly limited the scope of LHFE exemptions to WWII...

  20. Spawner-recruit relationships and fish stock carrying capacity in aquatic ecosystems

    DEFF Research Database (Denmark)

    MacKenzie, Brian; Myers, R.A.; Bowen, K.G.

    2003-01-01

    Few marine ecologists have addressed important questions about the relative productivity and carrying capacity of different ecosystems required to support fish populations. Whereas many researchers have investigated interannual variability in recruitment within a stock, we asked whether...

  1. Test-retest reliability of lifting and carrying in a 2-day functional capacity evaluation.

    Science.gov (United States)

    Reneman, M F; Dijkstra, P U; Westmaas, M; Göeken, L N H

    2002-12-01

    The objectives of this study were to establish test-retest reliability of lifting and carrying of a functional capacity evaluation (FCE) on two consecutive days and to verify the need for a 2-day protocol. A cohort of 50 patients (39 men, 11 women) with nonspecific low back pain were evaluated using a 2-day FCE protocol. Intraclass correlation coefficients (ICC) were calculated for weight lifted and carried. Predictive relationships between test and retest were explored by means of a regression analysis. The results of ICC were lifting low 0.87, lifting overhead 0.87, and carrying 0.77. Performances on day 2 were on an average 6-9% higher. Other than the amount of weight handled on day 1, no variable was found to predict performance on day 2. It was concluded that test-retest reliability of lifting and carrying was good, and the need for a 2-day protocol could not be confirmed.

  2. The importance of carrying a backpack in the rehabilitation of osteoporotic patients (biomechanical analysis).

    Science.gov (United States)

    Wendlova, J

    2011-01-01

    Based on a simple biomechanical analysis available to physicians, the article recommends carrying a backpack regularly as a part of the complex rehabilitation of osteoporotic patients. Carrying a backpack in front or on the back is recommended to patients with uncomplicated osteoporosis, while carrying a backpack on the back only is recommended to patients with osteporotic vertebral fractures. The importance of carrying a backpack is based upon removing the muscular dysbalance of the trunk muscles and upon increasing the bone strength by compressive force acting upon the vertebrae and proximal femur and activating osteoblasts to enhance the process of osteoformation. The backpack load is differentiated--patients with vertebral fractures put a weight up to 1 kg into their backpacks, patients without vertebral fractures increase the load up to 2 kg (Fig. 2, Ref. 12).

  3. Magnetic guide of cold atoms using a U-shaped current-carrying conductor

    Institute of Scientific and Technical Information of China (English)

    刘南春; 印建平

    2003-01-01

    We propose a new scheme to magnetically guide cold neutral atoms using a U-shaped current carrying conductor.The spatial magnetic field distribution from the U-shaped current-carrying conductor and the relationship between the magnetic field and parameters of the U-shaped conductor are analysed.Our study shows that U-shaped current-carrying conductor can be used to realize single-or double-channel magnetic guiding of cold atoms in weak-field-seeking states and to construct wrious atom-optical elements.By using Monte Carlo simulations,the dynamic process of the guided atomic-beam splitting in an atomic-beam splitter composed by the U-shaped current-carrying conductor is studied,and some results are presented.

  4. CARRIE web service: automated transcriptional regulatory network inference and interactive analysis.

    Science.gov (United States)

    Haverty, Peter M; Frith, Martin C; Weng, Zhiping

    2004-07-01

    We present an intuitive and interactive web service for CARRIE (Computational Ascertainment of Regulatory Relationships Inferred from Expression). CARRIE is a computational method that analyzes microarray and promoter sequence data to infer a transcriptional regulatory network from the response to a specific stimulus. This service displays an interactive graph of the inferred network and provides easy access to the evidence for the involvement of each gene in the network. We provide functionality to include network data in KEGG XML (KGML) format in this graph. Our service also provides Gene Ontology annotation to aid the user in forming hypotheses about the role of each gene in the cellular response. The CARRIE web service is freely available at http://zlab.bu.edu/CARRIE-web.

  5. Recreation carrying capacity estimations to support beach management at Praia de Faro, Portugal

    OpenAIRE

    Zacarias, D. A.; Williams, Allan T.; Newton, Alice

    2011-01-01

    The purpose of this research paper is to outline the theory and practice of tourism carrying capacity assessment and its relevance as a management tool for coastal management. Based on the Tourism Carrying Capacity Assessment for Protected Areas Framework and the Image Capture Technique associated with the PAOT (people at one time) approach, this paper explores Praia de Faro as the study area and attempts to assess the optimum number of people that should be allowed without jeopar...

  6. Heat Transfer Enhancement by Fluidized Solid Particles in Gas Carrying Evaporation

    Institute of Scientific and Technical Information of China (English)

    于志家; 孙成新; 孙相彧; 刘展红

    2001-01-01

    Heat transfer characteristics are studied for gas carrying evaporation with fluidized solid particles in a vertical rectangular conduit. Experimental results show that heat transfer of gas carrying evaporation is enhanced and the superheat of liquid in contact with heating surface lowers remarkably by introducing solid particles. Nucleate boiling on the heating surface is suppressed to a considerable degree. The mechanism of heat transfer enhancement by fluidized solid particles is analyzed with the consideration of collisions of solid particles with the boiling vapor bubbles.

  7. A 5-year audit of outcome of apicectomies carried out in a district general hospital.

    OpenAIRE

    Lyons, A. J.; Hughes, C E; Dixon, E. J.

    1995-01-01

    Success rates of up to 90% have been claimed for apicectomy. However, the conditions that this procedure is carried out under at district general hospitals may be at variance with such studies. A 5-year audit of outcome was therefore carried out within a district general hospital. It was found that 89% of apicected teeth still remained at 5 years. Outcome was not influenced by any of the factors examined, and could not be predicted radiographically. Most failures occurred after the average po...

  8. Analysis of Interpersonal Relationship in the Consumer Society Context in Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    张恩秀

    2009-01-01

    Carrie's characterization is a familiar topic in the criticism in Sister Carrie. However, how it could be re-viewed in the context of the consumer society is relalively a new issue. This paper attempts to analyze the interpersonal relationship in the context of a consumer society. It explores the consumer society's influences on the lovers and family. In addition, it points out Dreiser's ambivalent attitude towards consumer ideology.

  9. Theoretical Research of the Urban Comprehensive Carrying Capacity in the Epoch of Urbanization

    OpenAIRE

    Li Changliang; Lian Lina

    2012-01-01

    Conventional wisdom suggests that ecological factors are prominent constraints of cities in the urbanization process. By contrast, this article provides a fresh conception of the urban comprehensive carrying capacity in the context of ecosystem, economy and social dimensions. Based on in-depth analysis, the author makes out a list of informative area-based indicators in three differentiated layers and the findings are that the urban comprehensive carrying capacity is an entanglement of three ...

  10. Load-carrying capacity test of a long-span timber truss

    OpenAIRE

    Branco, Jorge M.; Varum, Humberto; Ramisote, Vitor; Costa, Aníbal

    2016-01-01

    The aim of this work was to study the behavior of existing timber trusses in order to improve the knowledge that may support engineers in the definition of more adequate restoration interventions in old buildings, particularly of long-span timber structures, as industrial or monumental constructions. For this purpose, a cyclic load-carrying test was carried out on a full-scale timber planar truss rescued from an old industrial roof structure. Before the tests, an accurate geometrical characte...

  11. Methicillin-resistant Staphylococcus saprophyticus in Sweden carries various types of staphylococcal cassette chromosome mec (SCCmec).

    Science.gov (United States)

    Söderquist, B; Berglund, C

    2009-12-01

    Staphylococcus saprophyticus is a common cause of uncomplicated urinary tract infections and is usually susceptible to the antimicrobial agents used for their treatment. However, S. saprophyticus resistant to beta-lactam antibiotics and carrying mecA has been reported. Eight Swedish isolates of mecA-positive S. saprophyticus with diverse origin carrying at least three different types of staphylococcal cassette chromosome mec (SCCmec) are described here.

  12. Evaluating Beijing's human carrying capacity from the perspective of water resource constraints.

    Science.gov (United States)

    Zhang, Yingxuan; Chen, Min; Zhou, Wenhua; Zhuang, Changwei; Ouyang, Zhiyun

    2010-01-01

    As the demands on limited water resources intensify, concerns are being raised about the human carrying capacity of these resources. However, few researchers have studied the carrying capacity of regional water resources. Beijing, the second-largest city in China, faces a critical water shortage that will limit the city's future development. We developed a method to quantify the carrying capacity of Beijing's water resources by considering water-use structures based on the proportions of water used for agricultural, industrial, and domestic purposes. We defined a reference structure as 45:22:33 (% of total, respectively), an optimized structure as 40:20:40, and an ideal structure as 50:15:35. We also considered four domestic water quotas: 55, 75, 95, and 115 m3/(person x yr). The urban carrying capacity of 10-12 million was closest to Beijing's actual 2003 population for all three water-use structures with urban domestic water use of 75 m3/(person x yr). However, after accounting for our underlying assumptions, the adjusted carrying capacity is closer to 5-6 million. Thus, Beijing's population in 2003 was almost twice the adjusted carrying capacity. Based on this result, we discussed the ecological and environmental problems created by Beijing's excessive population and propose measures to mitigate these problems.

  13. Sensitivity of the speed evaluation tests of carrying the ball in youth soccer players

    Directory of Open Access Journals (Sweden)

    Rakojević Bojan

    2016-01-01

    Full Text Available This research is aimed at examining sensitivity of the speed evaluation tests while carrying the ball. The research included 76 male examinees, aged 17 years (+/- 6 months, who were divided into two qualitatively different subgroups. For determining the speed while carrying the ball, the tests of M type of carrying the ball between cones and slalom ball carrying with a pass were performed. The obtained results proved that the examinees from the group of more successful soccer players (Group 1 scored better on the applied tests when compared to the results of the group consisting of the soccer players from lower ranked clubs and who are not national team members (Group 2. High values of the result homogeneity in the two groups (for the Groups 1 and 2 it was 75% and 68.75% respectively lead to the conclusion that this ability is essential characteristics of young soccer players. The coefficient of discrimination of 0.269 for the results of M type test and 0.197 for the results of the test of carrying the ball with a pass indicate that the results of the two tests provide possibility to qualitatively distinguish youth players. Therefore, it can be concluded that as a technical element, carrying the ball significantly affects the quality of players' performance.

  14. Estimation of Carrying Angle Based on CT Images in Preoperative Surgical Planning for Cubitus Deformities

    Directory of Open Access Journals (Sweden)

    Kim,Eugene

    2009-12-01

    Full Text Available Conventionally, the carrying angle of the elbow is measured using simple two-dimensional radiography or goniometry, which has questionable reliability. This study proposes a novel method for estimating carrying angles using computed tomography that can enhance the reliability of the angle measurement. Data of CT scans from 25 elbow joints were processed to build segmented three-dimensional models. The cross-sectional centerlines of the ulna and the humerus were traced from the 3D models, and the angle between 2 vectors formed from the centerlines of the humerus and the ulna was defined as the "three-dimensional carrying angle." These angles were compared with those measured by simple radiograph. Two cases of angular deformity were underwent surgery based on this preoperative surgical planning, and the postoperative 3D carrying angles were evaluated using the proposed method. The mean value of the calculated three-dimensional carrying angle was 20.7 degrees +/-3.61, while it was 16.3 degrees +/-3.21 based on simple radiography without statistical difference. Based on the 3D carrying angle estimations, 2 surgical cases of cubitus deformities were planned by comparison with the normal contra-lateral elbow. Postoperative angle estimations confirmed that the corrected angles were nearly identical to the planned angles for both cases. The results of this study showed that the carrying angle can be accurately estimated using three-dimensional CT and that the proposed method is useful in evaluating deformities of the elbow with high reliability.

  15. Spatial patterns of ecological carrying capacity supply-demand balance in China at county level

    Institute of Scientific and Technical Information of China (English)

    LIU Dong; FENG Zhiming; YANG Yanzhao; YOU Zhen

    2011-01-01

    A balanced ecological carrying capacity and its understanding are important to achieve sustainable development for human kind.Here,the concept of ecological carrying capacity has been used for measuring the dependencies between human and nature.China's ecological balance between supply and demand has become a global concern and is widely debated.In this study the 'Ecological Footprint' method was used to analyze the supply-demand balance of China's ecological carrying capacity.Firstly,the ecological supply and demand balance was calculated and evaluated,and secondly,the ecological carrying capacity index (ECCI) was derived for each county of China in 2007,and finally this paper systematically evaluated the ecological carrying capacity supply-demand balance of China.The results showed that ecological deficit appeared to be the main characteristic of ecological carrying capacity supply-demand balance in 2007 of China at county scale.In general,more than four-fifths of the Chinese population was concentrated in less than one-third of the land area and more than two-thirds of the land.area was inhabited by less than one-fifth of the population.The spatial distribution of the ecological carrying capacity demand-supply was unbalanced ranging from significant overloading to affluence from southeastern to northwestern part of China.It appeared to be more dominant in regions located at coastal areas which are attracted by migrants and had a generally higher population density.Along with the rapid development and urbanization trends in China,ecological deficits in these regions will become more severe.

  16. Why do three-wheelers carrying schoolchildren suffer very low fatal crashes?

    Directory of Open Access Journals (Sweden)

    Gaurav Pandey

    2015-03-01

    Full Text Available This paper investigates the possible causes of low fatalities in three-wheelers (autorickshaw carrying schoolchildren in India. The data was collected in the form of First Information Report (FIR from local police stations from 2007 to 2012 and video-graphic surveys were done on four arterial roads running through Ludhiana, Punjab, India. Surveys were also done on one subarterial road near school zone which was used by three-wheelers carrying schoolchildren. The objective of the study was to investigate the hypothesis that drivers behave differently while following or overtaking three-wheelers carrying children. Many researchers have investigated the effect of passengers on the driver of the same vehicle, but there was no evidence of any study which investigated the effect of child passengers on nearby vehicles. It was found that heavy vehicles maintain more gaps while following or overtaking three-wheelers carrying children as compared to those not carrying children. It was also found that this effect is more prominent at speeds higher than 40 km/h. On the other hand lighter vehicles keep the highest lateral and longitudinal gaps to heavy vehicles and three-wheelers without children respectively.

  17. A protocol for clinical evaluation of the carrying angle of the elbow by anatomic landmarks.

    Science.gov (United States)

    Zampagni, Maria Luisa; Casino, Daniela; Martelli, Sandra; Visani, Andrea; Marcacci, Maurilio

    2008-01-01

    The aim of this work was to present an in vivo protocol to estimate the carrying angle of the elbow in full extension. Forty-four arms were measured by using an electrogoniometer to acquire 3-dimensional coordinates of the landmarks. An algorithm based on the Cardan decomposition method was used to compute the carrying angle and the flexion and pronation angles of the elbow. The mean carrying angle was 12.42 degrees +/- 4.06 degrees , in agreement with the literature and with values obtained by a standard goniometer (r = 0.46; P = .000). Our protocol provided excellent repeatability (interclass correlation coefficient [ICC] = 0.85), greater than a goniometer (ICC = 0.76), and a standard error of measurement of only 1.62 degrees . Flexion was a significant factor (P = .01) in carrying angle estimation. This study suggests that the carrying angle cannot be estimated independently by the flexion angle, even when measured in apparently full extension, and it could be useful in elbow disorders, such as fractures or epicondylar disease management and evaluation of elbow reconstruction.

  18. Evaluation of Ecological Carrying Capacity of Henan Province under the Sustainable Development

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Based on the overview of social economy of Henan Province,I probe into the concept and evaluation of ecological carrying capacity.By using the ecological footprint analysis and the data of various kinds of land supply of Henan Province from 2000 to 2008,the ecological carrying capacity of Henan Province is analyzed.It is unveiled that inharmonious population,natural resources and economic resources affect the efficiency of the sustainable development of ecological carrying capacity of Henan Province;the underdeveloped economy of Henan Province impacts the sustainable development of ecological carrying capacity of Henan Province and the overburdened population lead to the insufficiency of ecological carrying capacity.Around protecting the threshold of ecological system,the countermeasures are put forward,which cover forming the idea of ecological economy and circular economy and promoting the transformation of economy growth mode;taking the development road of using resources intensively and performing the strategy of sustainable utilization of resources;strictly control population growth and strengthening people’s crisis awareness of resources and environment.

  19. Profile of women who carried out smoking cessation treatment: a systematic review

    Directory of Open Access Journals (Sweden)

    Caroline Figueira Pereira

    2015-01-01

    Full Text Available OBJECTIVE Analyze the profile of women, in health services, who carry out treatment for smoking cessation. METHODS Systematic review that used the following sources of information: Cummulative Index to Nursing and Allied Health Literature (CINAHL, PubMed, Biblioteca Virtual em Saúde (BVS, Scopus and Web of Science. We included quantitative studies that addressed the characterization of women, in health services, who carried out treatment for smoking cessation, resulting in 12 articles for analysis. The assessment of the methodological quality of the studies was performed using the instrument MAStARI from Joanna Briggs Institute. RESULTS The predominant profile of women who carried out treatment for smoking cessation in health services was composed of white, married, employed, and highly level educated women. Women who carried out the treatment for smoking cessation in specialized services had a more advanced age, were white, were married and had a diagnosis of depression. The quality level of most studies was moderate. CONCLUSIONS The profile of women who carry out treatment for smoking cessation, either in general or specialized health services, is composed of white, married, and highly level educated women. Publications about smoking women are scarce and the lack of Brazilian studies characterizing the profile of women who start treatment for smoking cessation shows the need for studies that explore this subject.

  20. Carrying capacity of U.S. agricultural land: Ten diet scenarios

    Directory of Open Access Journals (Sweden)

    Christian J. Peters

    2016-07-01

    Full Text Available Abstract Strategies for environmental sustainability and global food security must account for dietary change. Using a biophysical simulation model we calculated human carrying capacity under ten diet scenarios. The scenarios included two reference diets based on actual consumption and eight “Healthy Diet” scenarios that complied with nutritional recommendations but varied in the level of meat content. We considered the U.S. agricultural land base and accounted for losses, processing conversions, livestock feed needs, suitability of land for crops or grazing, and land productivity. Annual per capita land requirements ranged from 0.13 to 1.08 ha person-1 year-1 across the ten diet scenarios. Carrying capacity varied from 402 to 807 million persons; 1.3 to 2.6 times the 2010 U.S. population. Carrying capacity was generally higher for scenarios with less meat and highest for the lacto-vegetarian diet. However, the carrying capacity of the vegan diet was lower than two of the healthy omnivore diet scenarios. Sensitivity analysis showed that carrying capacity estimates were highly influenced by starting assumptions about the proportion of cropland available for cultivated cropping. Population level dietary change can contribute substantially to meeting future food needs, though ongoing agricultural research and sustainable management practices are still needed to assure sufficient production levels.

  1. Carrying weapons and intent to harm among Victorian secondary school students in 1999 and 2009.

    Science.gov (United States)

    Hemphill, Sheryl A; Tollit, Michelle A; Romaniuk, Helena; Williams, Joanne; Toumbourou, John W; Bond, Lyndal; Patton, George C

    2013-12-16

    To examine comparable survey data across 10 years to assess whether rates of self-reported weapon carrying and intent to harm others have increased as suggested in reported trends in violent offences. Population-based surveys administered to Victorian secondary school students in 1999 (8984 students) and 2009 (10 273 students) attending government, Catholic and independent schools. Student self-reports of carrying a weapon and attacking someone with the intent to harm in the past 12 months. In both surveys, about 15.0% of students reported carrying a weapon and about 7.0% reported attacking someone with intent to harm in the past 12 months, with higher rates among boys than girls. There was no change over time in the rates of students carrying weapons or attacking someone with the intent to harm, after controlling for demographic variables. In contrast to media portrayals and reported trends in violent offences, rates of students carrying weapons and attacking others with intent to harm have not changed between 1999 and 2009. These findings underline the importance of having national population-based data to regularly monitor the rates of these and related behaviours among young Australians.

  2. Variation in local carrying capacity and the individual fate of bacterial colonizers in the phyllosphere.

    Science.gov (United States)

    Remus-Emsermann, Mitja N P; Tecon, Robin; Kowalchuk, George A; Leveau, Johan H J

    2012-04-01

    Using a phyllosphere model system, we demonstrated that the term 'carrying capacity', as it is commonly used in microbial ecology, needs to be understood as the sum of many 'local carrying capacities' in order to better explain and predict the course and outcome of bacterial colonization of an environment. Using a green fluorescent protein-based bioreporter system for the quantification of reproductive success (RS) in individual Erwinia herbicola cells, we were able to reconstruct the contribution of individual immigrants to bacterial population sizes on leaves. Our analysis revealed that plant foliage represents to bacteria an environment where individual fate is determined by the local carrying capacity of the site where an immigrant cell lands. With increasing inoculation densities, the RS of most immigrants declined, suggesting that local carrying capacity under the tested conditions was linked to local nutrient availability. Fitting the observed experimental data to an adapted model of phyllosphere colonization indicated that there might exist three types of sites on leaves, which differ in their frequency of occurrence and local carrying capacity. Specifically, our data were consistent with a leaf environment that is characterized by few sites where individual immigrants can produce high numbers of offspring, whereas the remainder of the leaf offered an equal number of sites with low and medium RS. Our findings contribute to a bottom-up understanding of bacterial colonization of leaf surfaces, which includes a quantifiable role of chance in the experience at the individual level and in the outcome at the population level.

  3. La ascendencia paterna de Daniel Carrión García

    Directory of Open Access Journals (Sweden)

    Ricardo Álvarez

    2008-09-01

    Full Text Available Daniel Carrión García, mártir de la medicina peruana y héroe nacional, tenía antiguas raíces peruanas por línea materna, pero su ascendencia paterna era netamente ecuatoriana. Los Carrión fueron una de las familias más prominentes de la ciudad de Loja, a tal punto que, en 1830, al elegirse el primer Presidente de la República del Ecuador, uno de los dos candidatos fue don Manuel Carrión. La participación política de la familia continuó en los años siguientes y uno de sus miembros, el doctor José Baltazar Carrión y Torres, padre de Daniel, unió su destino al del General Juan José Flores, quien al caer en desgracia, luego de un frustrado intento revolucionario, lo arrastró consigo y debió exiliarse en el Perú. Nuestro objetivo es contribuir al conocimiento de la ascendencia paterna de Daniel Carrión García.

  4. Modelling of carrying capacity in National Park - Fruška Gora (Serbia case study

    Directory of Open Access Journals (Sweden)

    Vujko Aleksandra

    2017-03-01

    Full Text Available Negative effects of tourism development in a destination are usually the consequence of the high concentration of tourists, accommodation facilities and the activities that are practiced in a relatively restricted area. One of the most important measures to protect the areas is to calculate the maximum number of tourists that can simultaneously reside in a region, i.e. the determination of the carrying capacity. This paper outlines a method for determining carrying capacity based on zoning of environmental resources and zoning within a region. The paper argues for a return to the idea of identifying maximum appropriate number of users. The main hypothesis of the paper is based on the statement that the development of tourism in Fruška Gora (Mountain National Park in Northern Serbia must be in accordance with the basic principles of sustainability, including the determination of carrying capacity. The main research goal was to show the opinion of local residents about the uncontrolled development of tourism, and to determine the carrying capacity in four sports and recreational zones of the mountain. The carrying capacity of the area is calculated by Lavery and Stanev formulas.

  5. Profile of women who carried out smoking cessation treatment: a systematic review

    Science.gov (United States)

    Pereira, Caroline Figueira; de Vargas, Divane

    2015-01-01

    OBJECTIVE Analyze the profile of women, in health services, who carry out treatment for smoking cessation. METHODS Systematic review that used the following sources of information: Cummulative Index to Nursing and Allied Health Literature (CINAHL), PubMed, Biblioteca Virtual em Saúde (BVS), Scopus and Web of Science. We included quantitative studies that addressed the characterization of women, in health services, who carried out treatment for smoking cessation, resulting in 12 articles for analysis. The assessment of the methodological quality of the studies was performed using the instrument MAStARI from Joanna Briggs Institute. RESULTS The predominant profile of women who carried out treatment for smoking cessation in health services was composed of white, married, employed, and highly level educated women. Women who carried out the treatment for smoking cessation in specialized services had a more advanced age, were white, were married and had a diagnosis of depression. The quality level of most studies was moderate. CONCLUSIONS The profile of women who carry out treatment for smoking cessation, either in general or specialized health services, is composed of white, married, and highly level educated women. Publications about smoking women are scarce and the lack of Brazilian studies characterizing the profile of women who start treatment for smoking cessation shows the need for studies that explore this subject. PMID:26247386

  6. On Sister Carrie from Feminist Perspective and its Influence on Chinese Modern Women

    Institute of Scientific and Technical Information of China (English)

    毛武毅; 叶青

    2013-01-01

    Sister Carrie is a famous naturalistic novelist-Theodore Dreiser’s first novel in the first 20th century. It tells a story of a young rural woman Carrie how to become the red Broadway actress from the bottom of society, and the story vividly depicts the survival state and psychological and ideological process of Carrie. When it was published, it attracted attentions of critics from domestic and abroad at large, and they researched it from different perspectives. But most of their researches discussed from the materialism angle to concern with the expression of Determinism and Darwinism. So far, few people made the research from the topic of Carrie’s pursuit of self-achievement and perspective of feminist. Actually, not only did Carrie restrict on the material pursuit, but also boldly in pursuit of the self-achievement of new women. This paper aims at Carrie how to pursue the self-achievement of new women, and the influence of feminist to Chinese modern women.

  7. Profile of women who carried out smoking cessation treatment: a systematic review.

    Science.gov (United States)

    Pereira, Caroline Figueira; de Vargas, Divane

    2015-01-01

    OBJECTIVE Analyze the profile of women, in health services, who carry out treatment for smoking cessation. METHODS Systematic review that used the following sources of information: Cummulative Index to Nursing and Allied Health Literature (CINAHL), PubMed, Biblioteca Virtual em Saúde (BVS), Scopus and Web of Science. We included quantitative studies that addressed the characterization of women, in health services, who carried out treatment for smoking cessation, resulting in 12 articles for analysis. The assessment of the methodological quality of the studies was performed using the instrument MAStARI from Joanna Briggs Institute. RESULTS The predominant profile of women who carried out treatment for smoking cessation in health services was composed of white, married, employed, and highly level educated women. Women who carried out the treatment for smoking cessation in specialized services had a more advanced age, were white, were married and had a diagnosis of depression. The quality level of most studies was moderate. CONCLUSIONS The profile of women who carry out treatment for smoking cessation, either in general or specialized health services, is composed of white, married, and highly level educated women. Publications about smoking women are scarce and the lack of Brazilian studies characterizing the profile of women who start treatment for smoking cessation shows the need for studies that explore this subject.

  8. Comprehensive Measurement for Carrying Capacity of Resources and Environment of City Clusters in Central China

    Institute of Scientific and Technical Information of China (English)

    FANG Chuanglin; LIU Xiaoli

    2010-01-01

    Studying the carrying capacity of resources and environment of city clusters in the central China has impor-tant practical guidance significance for promoting the healthy,sustainable and stable development of this region.Ac-cording to their influencing factors and reciprocity mechanism,using system dynamics approaches,this paper built a SD model for measuring the carrying capacity of resources and environment of the city clusters in the central China,and through setting different development models,the comprehensive measurement analysis on the carrying capacity was carried out.The results show that the model of promoting socio-economic development under the protection of resources and environment is the optimal model for promoting the harmony development of resources,environment,society and economy in the city clusters.According to this model,the optimum population scale of the city clusters in2020 is 42.80×106 persons,and the moderate economic development scale is 22.055× 1012 yuan(RMB).In 1996-2020,the carrying capacity of resources and environment in the city clusters took on obvious phase-change characteristics.During the studied period,it is basically at the initial development stage,and will come through the development process from slow development to speedup development.

  9. The evolution of carrying capacity in constrained and expanding tumour cell populations.

    Science.gov (United States)

    Gerlee, Philip; Anderson, Alexander R A

    2015-08-12

    Cancer cells are known to modify their micro-environment such that it can sustain a larger population, or, in ecological terms, they construct a niche which increases the carrying capacity of the population. It has however been argued that niche construction, which benefits all cells in the tumour, would be selected against since cheaters could reap the benefits without paying the cost. We have investigated the impact of niche specificity on tumour evolution using an individual based model of breast tumour growth, in which the carrying capacity of each cell consists of two components: an intrinsic, subclone-specific part and a contribution from all neighbouring cells. Analysis of the model shows that the ability of a mutant to invade a resident population depends strongly on the specificity. When specificity is low selection is mostly on growth rate, while high specificity shifts selection towards increased carrying capacity. Further, we show that the long-term evolution of the system can be predicted using adaptive dynamics. By comparing the results from a spatially structured versus well-mixed population we show that spatial structure restores selection for carrying capacity even at zero specificity, which poses a solution to the niche construction dilemma. Lastly, we show that an expanding population exhibits spatially variable selection pressure, where cells at the leading edge exhibit higher growth rate and lower carrying capacity than those at the centre of the tumour.

  10. Modelling of carrying capacity in National Park - Fru\\vska Gora (Serbia) case study

    Science.gov (United States)

    Vujko, Aleksandra; Plavša, Jovan; Petrović, Marko D.; Radovanović, Milan; Gajić, Tamara

    2017-03-01

    Negative effects of tourism development in a destination are usually the consequence of the high concentration of tourists, accommodation facilities and the activities that are practiced in a relatively restricted area. One of the most important measures to protect the areas is to calculate the maximum number of tourists that can simultaneously reside in a region, i.e. the determination of the carrying capacity. This paper outlines a method for determining carrying capacity based on zoning of environmental resources and zoning within a region. The paper argues for a return to the idea of identifying maximum appropriate number of users. The main hypothesis of the paper is based on the statement that the development of tourism in Fru\\vska Gora (Mountain) National Park in Northern Serbia must be in accordance with the basic principles of sustainability, including the determination of carrying capacity. The main research goal was to show the opinion of local residents about the uncontrolled development of tourism, and to determine the carrying capacity in four sports and recreational zones of the mountain. The carrying capacity of the area is calculated by Lavery and Stanev formulas.

  11. Categorical and continuous - disentangling the neural correlates of the carry effect in multi-digit addition

    Directory of Open Access Journals (Sweden)

    Dressel Katharina

    2010-11-01

    Full Text Available Abstract Background Recently it was suggested that the carry effect observed in addition involves both categorical and continuous processing characteristics. Methods In the present study, we aimed at identifying the specific neural correlates associated with processing either categorical or continuous aspects of the carry effect in an fMRI study on multi-digit addition. Results In line with our expectations, we observed two distinct parts of the fronto-parietal network subserving numerical cognition to be associated with either one of these two characteristics. On the one hand, the categorical aspect of the carry effect was associated with left-hemispheric language areas and the basal ganglia probably reflecting increased demands on procedural and problem solving processes. Complementarily, the continuous aspect of the carry effect was associated with increased intraparietal activation indicating increasing demands on magnitude processing as well as place-value integration with increasing unit sum. Conclusions In summary, the findings suggest representations and processes underlying the carry effect in multi-digit addition to be more complex and interactive than assumed previously.

  12. Carrying capacities for nature parks as engines for sustainable regional development

    DEFF Research Database (Denmark)

    Brandt, Jesper; Holmes, Esbern

    of national and nature parks. Nature parks should rather develop towards a status as 'Biosphere Reserves' representing models for regional development, land use and landscape design in general. A relevant zoning of the regional environment of the Parks will be included as an instrument for such a strategy.......Growth in the number of visitors is an upcoming problem in nature parks. Nature parks are at the same time facing increasing demand, falling public appropriations and receding focus on their conservation functions. To ensure a balancing of nature protection and economic utilization the concept...... of carrying capacity has received increasing attention among park-authorities all over the world. A comparative analysis of conditions and initiatives related to visitor/nature carrying capacities in 8 nature parks in the Baltic region has been carried out. All the parks are candidates for recognition...

  13. An Alternative Approach for High Speed Railway Carrying Capacity Calculation Based on Multiagent Simulation

    Directory of Open Access Journals (Sweden)

    Mo Gao

    2016-01-01

    Full Text Available It is a multiobjective mixed integer programming problem that calculates the carrying capacity of high speed railway based on mathematical programming method. The model is complex and difficult to solve, and it is difficult to comprehensively consider the various influencing factors on the train operation. The multiagent theory is employed to calculate high speed railway carrying capacity. In accordance with real operations of high speed railway, a three-layer agent model is developed to simulate the operating process of high speed railway. In the proposed model, railway network agent, line agent, station agent, and train agent are designed, respectively. To validate the proposed model, a case study is performed for Beijing–Shanghai high speed railway by using NetLogo software. The results are consistent with the actual data, which implies that the proposed multiagent method is feasible to calculate the carrying capacity of high speed railway.

  14. Introducing carrying capacity-based normalisation in LCA: framework and development of references at midpoint level

    DEFF Research Database (Denmark)

    Bjørn, Anders; Hauschild, Michael Zwicky

    2015-01-01

    capacity of a reference region divided by its population and thus describes the annual personal share of the carrying capacity.The developed references can be applied to indicator results obtained using commonly applied characterisation models in LCIA. The European NR are generally lower than the global NR...... ozone formation and soil quality were found to exceed carrying capacities several times.The developed carrying capacity-based normalisation references offer relevant supplementary reference information to the currently applied references based on society’s background interventions by supporting...... an evaluation of the environmental sustainability of product systems on an absolute scale.Challenges remain with respect to spatial variations to increase the relevance of the normalisation references for impact categories that function at the local or regional scale. The sensitivity of NR to different choices...

  15. Siblings and Adolescent Weapon Carrying: Contributions of Genetics, Shared Environment, and Nonshared Environment.

    Science.gov (United States)

    Wallace, Lacey N

    2017-07-01

    Many past studies have observed evidence of sibling similarity and influence for delinquency and substance use. However, studies of sibling similarity for adolescent weapon carrying, particularly for weapons beyond firearms, are largely absent from the literature. The present study assesses sibling similarity in weapon carrying as well as the relative contributions of genetics, shared environment, and nonshared environment. Data are obtained from the first two waves of the National Longitudinal Study of Adolescent to Adult Health and analyzed using biometrical genetic models for twins and actor-partner interdependence models for nontwins. Results indicate little, if any, contribution stemming from genetics. There is also no evidence of a significant shared environment effect. Instead, all or nearly all of the variation and similarity in weapon carrying among siblings are related to the nonshared environment, particularly gang affiliation. Implications and possible extensions of these findings are discussed.

  16. First Identification of a Patient Colonized With Klebsiella pneumoniae Carrying blaNDM-1 in Taiwan

    Directory of Open Access Journals (Sweden)

    Hua-Shin Wu

    2010-11-01

    Full Text Available New Delhi metallo-β-lactamase 1 (NDM-1 is a novel type of metallo-β-lactamase (MBL. Enterobacteriaceae carrying this NDM-1 encoding gene, blaNDM-1, have been identified worldwide. Bacteria carrying blaNDM-1 are not only resistant to carbapenem, but also highly resistant to many classes of antibiotics, which indicate the importance of prompt identification of these bacteria and implementation of strict infection control measures to prevent their transmission. Here, we report the first identification and management of a patient colonized with Klebsiella pneumoniae carrying blaNDM-1 in Taiwan, who returned from New Delhi where he had been hospitalized for a gun-shot injury.

  17. Biophysical and RNA Interference Inhibitory Properties of Oligonucleotides Carrying Tetrathiafulvalene Groups at Terminal Positions

    Directory of Open Access Journals (Sweden)

    Sónia Pérez-Rentero

    2013-01-01

    Full Text Available Oligonucleotide conjugates carrying a single functionalized tetrathiafulvalene (TTF unit linked through a threoninol molecule to the 3′ or 5′ ends were synthesized together with their complementary oligonucleotides carrying a TTF, pyrene, or pentafluorophenyl group. TTF-oligonucleotide conjugates formed duplexes with higher thermal stability than the corresponding unmodified oligonucleotides and pyrene- and pentafluorophenyl-modified oligonucleotides. TTF-modified oligonucleotides are able to bind to citrate-stabilized gold nanoparticles (AuNPs and produce stable gold AuNPs functionalized with oligonucleotides. Finally, TTF-oligoribonucleotides have been synthesized to produce siRNA duplexes carrying TTF units. The presence of the TTF molecule is compatible with the RNA interference mechanism for gene inhibition.

  18. Estimating the carrying capacity of green mussel cultivation by using net nutrient removal model.

    Science.gov (United States)

    Srisunont, Chayarat; Babel, Sandhya

    2016-11-15

    This study aims to evaluate the nutrient removal potential and carrying capacity of green mussel cultivation by using the mass balance model. The developed model takes into consideration the green mussel growth rate, density and chlorophyll a concentration. The data employed in this study were based on culture conditions at Sriracha Fisheries Research Station, Thailand. Results show that net nutrient removal by green mussel is 3302, 380, and 124mg/year/indv for carbon, nitrogen, and phosphorus respectively. The carrying capacity of green mussel cultivation was found to be 300indv/m(2) based on chlorophyll a concentration which will not release phosphorus in the water environment beyond the standard (45μg-PO4(-3)-P/L). Higher chlorophyll a concentration results in lowered green mussel carrying capacity. This model can assist farm operators with possible management strategies for a sustainable mussel cultivation and protection of the marine environment.

  19. A 5-year audit of outcome of apicectomies carried out in a district general hospital.

    Science.gov (United States)

    Lyons, A. J.; Hughes, C. E.; Dixon, E. J.

    1995-01-01

    Success rates of up to 90% have been claimed for apicectomy. However, the conditions that this procedure is carried out under at district general hospitals may be at variance with such studies. A 5-year audit of outcome was therefore carried out within a district general hospital. It was found that 89% of apicected teeth still remained at 5 years. Outcome was not influenced by any of the factors examined, and could not be predicted radiographically. Most failures occurred after the average postoperative review period of 10.5 months. Patient satisfaction with the procedure was high at over 90%. Based on these results it was concluded that apicectomy was an effective procedure when carried out by staff of all grades within the district general hospital, and that repeated follow-up appointments with radiographs over the first postoperative year were not useful. PMID:7574319

  20. Conditions for the management of carrying capacity in the parks of Parks&Benefits

    DEFF Research Database (Denmark)

    Brandt, Jesper; Holmes, Esbern

    2011-01-01

    Growth in the number of visitors is an upcoming problem in nature parks. Nature parks are at the same time facing increasing demand, falling public appropriations and receding focus on their conservation functions. To ensure a balancing of nature protection and economic utilization the concept...... of carrying capacity has received increasing attention among park-authorities all over the world. Carrying capacities understood as limits or standards not to be exceeded to protect a supporting landscape system are not scientifically determined sizes. They are a result of political decision processes among...... stakeholders, balancing use and protection preferably based on scientific and/or experiential cognition. The conditions for the management of carrying capacity for the 8 nature parks in the EC Baltic Project Parks&Benefits are analysed in the report. 1. Part focus on the methodology, concentrated...

  1. Threat by association: do distant intergroup threats carry-over into local intolerance?

    Science.gov (United States)

    Bouman, Thijs; van Zomeren, Martijn; Otten, Sabine

    2014-09-01

    Individuals are often confronted with intergroup threats, yet many of these threats emanate from distant groups that most individuals are unlikely to encounter in their local environment. An important yet unanswered question is whether reactions to those threats, such as intolerance towards the threatening group, carry over to other groups that individuals actually do encounter in their local environment (e.g., immigrants). The main goal of our studies was to experimentally identify this carry-over effect of intergroup threat. Specifically, we hypothesized that (by definition relatively abstract) symbolic threats (e.g., threats to the ingroup's worldview) have an especially strong carry-over potential because those threats can be easily attributed to other outgroups. We tested these predictions in one correlational and two experimental studies. The results of all three studies confirmed our hypothesis that particularly distant symbolic threats were predictive of intolerance towards local outgroups.

  2. Missile Captive Carry Monitoring and Helicopter Identification Using a Capacitive Microelectromechanical Systems Accelerometer

    Energy Technology Data Exchange (ETDEWEB)

    Hatchell, Brian K.; Mauss, Fredrick J.; Amaya, Ivan A.; Skorpik, James R.; Silvers, Kurt L.; Marotta, Steve

    2012-03-27

    Military missiles are exposed to many sources of mechanical vibration that can affect system reliability, safety, and mission effectiveness. The U. S. Army Aviation and Missile Research Development and Engineering Center (AMRDEC) has been developing missile health monitoring systems to assess and improve reliability, reduce life cycle costs, and increase system readiness. One of the most significant exposures to vibration occurs when the missile is being carried by a helicopter or other aviation platform, which is a condition known as captive carry. Recording the duration of captive carry exposure during the missile’s service life can enable the implementation of predictive maintenance and resource management programs. Since the vibration imparted by each class of helicopter varies in frequency and amplitude, tracking the vibration exposure from each helicopter separately can help quantify the severity and harmonic content of the exposure. Under the direction of AMRDEC staff, engineers at the Pacific Northwest National Laboratory have developed a Captive Carry Health Monitor (CCHM) for the Hellfire II missile. The CCHM is an embedded usage monitoring device installed on the outer skin of the Hellfire II missile to record the cumulative hours the host missile has been in captive carry mode. To classify the vibration by class of helicopter, the CCHM analyzes the amplitude and frequency content of the vibration with the Goertzel algorithm to detect the presence of distinctive rotor harmonics. Cumulative usage data are accessible in theater from an external display; monthly usage histograms are accessible through an internal download connector. This paper provides an overview of the CCHM electrical and package design, describes field testing and data analysis techniques used to monitor captive carry identify and the class of helicopter, and discusses the potential application of missile health and usage data for real-time reliability analysis and fleet management.

  3. Performance of women with fibromyalgia in walking up stairs while carrying a load

    Directory of Open Access Journals (Sweden)

    Daniel Collado-Mateo

    2016-02-01

    Full Text Available Background. Fibromyalgia is a chronic disease characterized by widespread pain and other associated symptoms. It has a relevant impact on physical fitness and the ability to perform daily living tasks. The objective of the study was to analyze the step-by-step-performance and the trunk tilt of women with fibromyalgia in the 10-step stair climbing test compared with healthy controls. Methods. A cross-sectional study was carried out. Twelve women suffering from fibromyalgia and eight healthy controls were recruited from a local association. Participants were asked to climb 10 stairs without carrying a load and 10 stairs carrying a load of 5 kg in each hand. Mediolateral trunk tilt was assessed using the “Functional Assessment of Biomechanics (FAB” wireless motion capture device, and the time between steps was assessed via weight-bearing insoles. Results. Trunk tilt in the stair-climbing task carrying a load was significantly higher in women with fibromyalgia when compared to the healthy controls (2.31 (0.63 vs. 1.69 (0.51 respectively. The effect of carrying a load was significantly higher for women with fibromyalgia compared with healthy controls at the intermediate and final part of the task. Discussion. Trunk tilt during stair climbing while carrying a load was higher in women with FM, which could increase the risk of falling. Additionally, women with FM experienced a higher pace slowdown as a consequence of the load, which supports the need of including specific strength and resistance training to physical therapies for this population.

  4. Performance of women with fibromyalgia in walking up stairs while carrying a load.

    Science.gov (United States)

    Collado-Mateo, Daniel; Adsuar, José C; Olivares, Pedro R; Dominguez-Muñoz, Francisco J; Maestre-Cascales, Cristina; Gusi, Narcis

    2016-01-01

    Background. Fibromyalgia is a chronic disease characterized by widespread pain and other associated symptoms. It has a relevant impact on physical fitness and the ability to perform daily living tasks. The objective of the study was to analyze the step-by-step-performance and the trunk tilt of women with fibromyalgia in the 10-step stair climbing test compared with healthy controls. Methods. A cross-sectional study was carried out. Twelve women suffering from fibromyalgia and eight healthy controls were recruited from a local association. Participants were asked to climb 10 stairs without carrying a load and 10 stairs carrying a load of 5 kg in each hand. Mediolateral trunk tilt was assessed using the "Functional Assessment of Biomechanics (FAB)" wireless motion capture device, and the time between steps was assessed via weight-bearing insoles. Results. Trunk tilt in the stair-climbing task carrying a load was significantly higher in women with fibromyalgia when compared to the healthy controls (2.31 (0.63) vs. 1.69 (0.51) respectively). The effect of carrying a load was significantly higher for women with fibromyalgia compared with healthy controls at the intermediate and final part of the task. Discussion. Trunk tilt during stair climbing while carrying a load was higher in women with FM, which could increase the risk of falling. Additionally, women with FM experienced a higher pace slowdown as a consequence of the load, which supports the need of including specific strength and resistance training to physical therapies for this population.

  5. Correlates of weapon carrying among high school students in the United States

    Science.gov (United States)

    Muula, Adamson S; Rudatsikira, Emmanuel; Siziya, Seter

    2008-01-01

    Background Deaths and injuries arising from interpersonal violence among adolescents are major public health concerns in the United States. The bearing of weapons among adolescents is a critical factor in many of these deaths and injuries. Methods A secondary analysis of the 2005 United States Youth Risk Behavior Surveillance System Survey data was carried out to examine the variables associated with self-reported history of weapon carrying on school property among high school students. We used logistic regression analysis to assess the associations. Results Of the 13,707 respondents who participated in the survey, 10.2% of males and 2.6% of females reported carrying a weapon on school property. In multivariate logistic regression analysis, males were more likely to report having carried a weapon than females (odds ratio (OR) = 5.58; 95% confidence interval (CI) [4.23, 7.62]). Self-reported race/ethnicity was also associated with weapon carrying. Other variables positively associated with weapon carrying at school were substance use (OR = 1.77; 95% CI [1.16, 2.68]), depression (OR = 1.44; 95% CI [1.10, 1.89]), suicidal ideation (OR = 1.64; 95% CI [1.23, 2.19]), having had property stolen or deliberately damaged at school (OR = 1.55; 95% CI [1.21, 1.98]), having been raped (OR = 1.70; 95% CI [1.22, 2.37]), having been threatened or injured with a weapon on school property (OR = 2.19; 95% CI [1.63, 2.95]), and having engaged in physical fighting (OR = 2.02; 95% CI [1.56, 2.63]). Conclusion This research identifies factors that are associated with weapon bearing among adolescents in the United States. These factors may be important in the design of interventions aimed at improving school safety and adolescent health. PMID:18605995

  6. Performance of women with fibromyalgia in walking up stairs while carrying a load

    Science.gov (United States)

    Adsuar, José C.; Olivares, Pedro R.; Dominguez-Muñoz, Francisco J.; Maestre-Cascales, Cristina; Gusi, Narcis

    2016-01-01

    Background. Fibromyalgia is a chronic disease characterized by widespread pain and other associated symptoms. It has a relevant impact on physical fitness and the ability to perform daily living tasks. The objective of the study was to analyze the step-by-step-performance and the trunk tilt of women with fibromyalgia in the 10-step stair climbing test compared with healthy controls. Methods. A cross-sectional study was carried out. Twelve women suffering from fibromyalgia and eight healthy controls were recruited from a local association. Participants were asked to climb 10 stairs without carrying a load and 10 stairs carrying a load of 5 kg in each hand. Mediolateral trunk tilt was assessed using the “Functional Assessment of Biomechanics (FAB)” wireless motion capture device, and the time between steps was assessed via weight-bearing insoles. Results. Trunk tilt in the stair-climbing task carrying a load was significantly higher in women with fibromyalgia when compared to the healthy controls (2.31 (0.63) vs. 1.69 (0.51) respectively). The effect of carrying a load was significantly higher for women with fibromyalgia compared with healthy controls at the intermediate and final part of the task. Discussion. Trunk tilt during stair climbing while carrying a load was higher in women with FM, which could increase the risk of falling. Additionally, women with FM experienced a higher pace slowdown as a consequence of the load, which supports the need of including specific strength and resistance training to physical therapies for this population. PMID:26855878

  7. A Novel Evaluation algorithm of Urban Comprehensive Carrying Capacity Index System in Northwest China:Lanzhou city as a example

    Science.gov (United States)

    Li, Jing; Lei, Song; Ling, Ma

    2016-06-01

    With the rapid growth of the urban population and quick expansion of urban dimensions, the affecting urban development constraints of urban carrying capacity has increasingly been attracting people's attention. However, the effective synthesization of urban resources carrying capacity, urban environmental carrying capacity, economic carrying capacity, society carrying capacity and traffic carrying capacity hardly acted as an urban comprehensive carrying capacity evaluation system. In this paper, the novel comprehensive index system of urban resources carrying capacity, urban environmental carrying capacity, economic carrying capacity, society carrying capacity and traffic carrying capacity was used for analyzing and evaluating urban comprehensive carrying capacity. Firstly, the standardization of evaluation index was constructed in the urban comprehensive carrying capacity index system. In addition, the past decade data from 2002 to 2011 was analyzed and evaluated in the weighted index system. Finally, the influence mechanism of various carrying capacity factors on urban comprehensive carrying capacity was investigated in Lanzhou. Experimental results can be summarized that the variation tendency of urban comprehensive carrying capacity was the annual increase of levels and the promotion countermeasure was put forward with congenital restricting factors and acquired restricting factors.

  8. Notification on carrying out the special rectification of rare earth production order nationwide

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    To implement "Some Opinions of the State Council of P.R.C. on Promoting the Sustainable and Healthy Development of Rare Earth Industry", regulate and standardize rare earth production order and accelerate the transformation of rare earth industry development pattern, the Ministry of Industry and Information Technology, Ministry of Supervision, Ministry of Environmental Protection, the State Administration of Taxation, the State Administration for Industry and Commerce and the State Administration of Work Safety jointly issued the "Notification on Carrying out the Special Rectification of Rare Earth Production Order Nationwide" and decided to carry out the special campaigns from 2011 August 1 to December 31.

  9. Fault Tolerant Variable Block Carry Skip Logic (VBCSL) using Parity Preserving Reversible Gates

    CERN Document Server

    Islam, Md Saiful; Begum, Zerina; Hafiz, Mohd Zulfiquar

    2010-01-01

    Reversible logic design has become one of the promising research directions in low power dissipating circuit design in the past few years and has found its application in low power CMOS design, digital signal processing and nanotechnology. This paper presents the efficient design approaches of fault tolerant carry skip adders (FTCSAs) and compares those designs with the existing ones. Variable block carry skip logic (VBCSL) using the fault tolerant full adders (FTFAs) has also been developed. The designs are minimized in terms of hardware complexity, gate count, constant inputs and garbage outputs. Besides of it, technology independent evaluation of the proposed designs clearly demonstrates its superiority with the existing counterparts.

  10. On the major Characteristics of Writing in Dreiser’s Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    张含

    2014-01-01

    Dreiser’s Sister Carrie has been controverted for a long time. It is still consummate today and as alive and valid as when it was written. First, the description of the psychology plays an important role in revealing the personalities of the major characters and deepens the theme. The second is Dreiser’s usage of comparison. Finally Dreiser uses the method of realism like Balzac. Sister Carrie is the exact reflection and the mirror of the society. Its special characteristics and the realistic writing style start a new field in American literature.

  11. Surface barrier effect on magnetization of a current-carrying type-II superconductor

    Science.gov (United States)

    Genenko, Yu. A.; Freyhardt, H. C.

    2000-10-01

    Vortex penetration in and exit from ideal flux-filled current-carrying superconductors are studied in the frame of the modified Meissner state model accounting for the vortex interactions with the sample surface, transport current, external field and the other vortices inside the sample. The generalized Silsbee's rule is first formulated and the M- H diagram of ideal current-carrying superconductors in a parallel field is estimated which is irreversible due to the surface effect alone. In general, magnetic behaviour of superconductors is shown to be more reversible in the presence of a transport current.

  12. Clonal spread of blaOXA-72-carrying Acinetobacter baumannii sequence type 512 in Taiwan.

    Science.gov (United States)

    Kuo, Han-Yueh; Hsu, Po-Jui; Chen, Jiann-Yuan; Liao, Po-Cheng; Lu, Chia-Wei; Chen, Chang-Hua; Liou, Ming-Li

    2016-07-01

    This is the first report to show an insidious outbreak of armA- and blaOXA-72-carrying Acinetobacter baumannii sequence type 512 (ST512) at a study hospital in northern Taiwan. Multilocus sequence typing revealed that this was a ST512 clone. All of the isolates with ST512 carried a novel 12,056-bp repGR2 in combination with a repGR12-type plasmid. This plasmid, designated pAB-ML, had one copy of the blaOXA-72 gene that was flanked by XerC/XerD-like sites and conferred resistance to carbapenems.

  13. Designing of Modified Area Efficient Square Root Carry Select Adder(SQRT CSLA)

    OpenAIRE

    Priya Meshram; Prof.Mamta Sarode

    2015-01-01

    In the design of Integrated Circuits, The necessity of portable systems is increasing an area occupancy plays a vital role. Square Root Carry Select Adder (SQRT CSLA) is one of the fastest adders which is used in this data-processing processor to perform fast arithmetic functions. In this paper, an area-efficient square root carry select adder(SQRT CSLA design) by sharing Common Boolean logic term (CBL) is proposed The modified architecture has been developed using Binary to Excess-1 converte...

  14. Design of Modified Area Efficient Square Root Carry Select Adder (SQRT CSLA)

    OpenAIRE

    Priya Meshram; Prof.Mamta Sarode

    2015-01-01

    In the design of Integrated Circuits, The necessity of portable systems is increasing an area occupancy plays a vital role. Square Root Carry Select Adder (SQRT CSLA) is one of the fastest adders which is used in this data-processing processor to perform fast arithmetic functions. In this paper, an area-efficient square root carry select adder(SQRT CSLA design) by sharing Common Boolean logic term (CBL) is proposed The modified architecture has been developed using Binary to Excess-1 converte...

  15. Cuttings-carried theory and erosion rule in gas drilling horizontal well

    Directory of Open Access Journals (Sweden)

    Wei Na

    2014-01-01

    Full Text Available In gas horizontal drilling, the gas with cuttings will go through the annulus at high speed which will lead strong erosion to the drill tools. This paper proposes a cuttings-carried theory and modified the critical cuttings-carried model for the gas-solid flow. Meanwhile, the erosive energy is obtained through simulating the gas-solid mixture in different conditions. The study result has positive significance on the determination of reasonable injection volume by optimizing construction parameters of horizontal well in gas drilling.

  16. On the first tachinid fly (Diptera, Tachinidae carrying Asclepiadoideae pollinaria in the Neotropical Region

    Directory of Open Access Journals (Sweden)

    Silvio Shigueo Nihei

    2011-09-01

    Full Text Available On the first tachinid fly (Diptera, Tachinidae carrying Asclepiadoideae pollinaria in the Neotropical Region. This paper reports the first Neotropical Tachinidae species possibly associated to pollination of Asclepiadoideae: a female of Euacaulona sumichrasti Townsend, 1908 (Diptera, Tachinidae, Phasiinae, Trichopodini carrying pollinaria of Gonolobus parviflorus Decne., 1844 (Apocynaceae, Asclepiadoideae, Asclepiadeae: Gonolobinae attached to its proboscis. The fly specimen was collected in Paraguay, Departamento Canindeyú. The pollinarium is illustrated and described herein. This represents the first anthophilous record to G. parviflorus and to the genus.

  17. Low Power Modulo 2n+1 Adder Based on Carry Save Diminished-One Number System

    Directory of Open Access Journals (Sweden)

    Somayeh Timarchi

    2008-01-01

    Full Text Available Modulo 2n+1 adders find great applicability in several applications including RNS implementations. This paper presents a new number system called Carry Save Diminished-one for modulo 2n+1 addition and a novel addition algorithm for its operands. In this paper, we also present a novel architectures for designing modulo 2n+1 adders, based on parallel-prefix carry computation units. CMOS implementations reveal the superiority of the resulting adders against previously reported solutions in terms of implementation area and delay.

  18. The Conflict Between the Traditional Value and Reality In Sister Carrie

    Institute of Scientific and Technical Information of China (English)

    顾庆媛; 陈飞; 孙胜强; 常利娜; 张罗斌

    2009-01-01

    Theodore Dreiser relates a story of Carrie's rise from a poor country girl to a Broadway star in Sister Carrie.An attempt is rnade in this paper to analyze the cold society.Byway of analyzing this unequal and cruel reality to prove:in the world only the fittest can survive.the standards and requirements of morality are challenged and the traditional value is suspected.In the cold reality.any way that Can protect oneself or survivein the wodd is sensible,morality and traditional value are weak and mcaningless.

  19. THE ENTERPRISE - THE LEGAL FORM FOR CARRYING ON AN ACTIVITY HAVING A PROFESSIONAL NATURE

    Directory of Open Access Journals (Sweden)

    Stanciu D. C RPENARU

    2011-08-01

    Full Text Available The new Romanian Civil Code has institutionalised a new conception regarding the regulating system for civil and commercial legal relations. Thus, new concepts emerge to fit the new conception, concepts regarding to the persons, the professionals, and the carrying on of an organised and systematised activity that qualifies such activity as having a professional nature. As one can find the new civil code has changed the conception regarding the enterprise, as it resulted from the actual commercial code. The operation of an enterprise will represent the legal form of carrying on an activity having a professional nature.

  20. Carrying capacity for the electromagnetic suspension low-speed maglev train on the horizontal curve

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Aiming at the lateral dislocation between the electromagnets and the rails on a horizontal curve,we investigated a single magnetic bogie of the maglev train in this paper.The magnetic levitation and guidance forces supplied by the suspension modules were deduced by the flux tube method.According to the dynamic equilibrium equations of the maglev train on the curved track with cant,several major factors that influence the carrying capacity were analyzed,and the formula of the carrying capacity was presented.The results provide a theoretical reference for the design of maglev train.