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Sample records for bacterial inclusion bodies

  1. Bacterial Inclusion Bodies: Discovering Their Better Half.

    Science.gov (United States)

    Rinas, Ursula; Garcia-Fruitós, Elena; Corchero, José Luis; Vázquez, Esther; Seras-Franzoso, Joaquin; Villaverde, Antonio

    2017-09-01

    Bacterial inclusion bodies (IBs) are functional, non-toxic amyloids occurring in recombinant bacteria showing analogies with secretory granules of the mammalian endocrine system. The scientific interest in these mesoscale protein aggregates has been historically masked by their status as a hurdle in recombinant protein production. However, progressive understanding of how the cell handles the quality of recombinant polypeptides and the main features of their intriguing molecular organization has stimulated the interest in inclusion bodies and spurred their use in diverse technological fields. The engineering and tailoring of IBs as functional protein particles for materials science and biomedicine is a good example of how formerly undesired bacterial byproducts can be rediscovered as promising functional materials for a broad spectrum of applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Towards revealing the structure of bacterial inclusion bodies.

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    Wang, Lei

    2009-01-01

    Protein aggregation is a widely observed phenomenon in human diseases, biopharmaceutical production, and biological research. Protein aggregates are generally classified as highly ordered, such as amyloid fibrils, or amorphous, such as bacterial inclusion bodies. Amyloid fibrils are elongated filaments with diameters of 6-12 nm, they are comprised of residue-specific cross-beta structure, and display characteristic properties, such as binding with amyloid-specific dyes. Amyloid fibrils are associated with dozens of human pathological conditions, including Alzheimer disease and prion diseases. Distinguished from amyloid fibrils, bacterial inclusion bodies display apparent amorphous morphology. Inclusion bodies are formed during high-level recombinant protein production, and formation of inclusion bodies is a major concern in biotechnology. Despite of the distinctive morphological difference, bacterial inclusion bodies have been found to have some amyloid-like properties, suggesting that they might contain structures similar to amyloid-like fibrils. Recent structural data further support this hypothesis, and this review summarizes the latest progress towards revealing the structural details of bacterial inclusion bodies.

  3. Isolation of cell-free bacterial inclusion bodies.

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    Rodríguez-Carmona, Escarlata; Cano-Garrido, Olivia; Seras-Franzoso, Joaquin; Villaverde, Antonio; García-Fruitós, Elena

    2010-09-17

    Bacterial inclusion bodies are submicron protein clusters usually found in recombinant bacteria that have been traditionally considered as undesirable products from protein production processes. However, being fully biocompatible, they have been recently characterized as nanoparticulate inert materials useful as scaffolds for tissue engineering, with potentially wider applicability in biomedicine and material sciences. Current protocols for inclusion body isolation from Escherichia coli usually offer between 95 to 99% of protein recovery, what in practical terms, might imply extensive bacterial cell contamination, not compatible with the use of inclusion bodies in biological interfaces. Using an appropriate combination of chemical and mechanical cell disruption methods we have established a convenient procedure for the recovery of bacterial inclusion bodies with undetectable levels of viable cell contamination, below 10⁻¹ cfu/ml, keeping the particulate organization of these aggregates regarding size and protein folding features. The application of the developed protocol allows obtaining bacterial free inclusion bodies suitable for use in mammalian cell cultures and other biological interfaces.

  4. Amyloid-linked cellular toxicity triggered by bacterial inclusion bodies

    International Nuclear Information System (INIS)

    Gonzalez-Montalban, Nuria; Villaverde, Antonio; Aris, Anna

    2007-01-01

    The aggregation of proteins in the form of amyloid fibrils and plaques is the characteristic feature of some pathological conditions ranging from neurodegenerative disorders to systemic amyloidoses. The mechanisms by which the aggregation processes result in cell damage are under intense investigation but recent data indicate that prefibrillar aggregates are the most proximate mediators of toxicity rather than mature fibrils. Since it has been shown that prefibrillar forms of the nondisease-related misfolded proteins are highly toxic to cultured mammalian cells we have studied the cytoxicity associated to bacterial inclusion bodies that have been recently described as protein deposits presenting amyloid-like structures. We have proved that bacterial inclusion bodies composed by a misfolding-prone β-galactosidase fusion protein are clearly toxic for mammalian cells but the β-galactosidase wild type enzyme forming more structured thermal aggregates does not impair cell viability, despite it also binds and enter into the cells. These results are in the line that the most cytotoxic aggregates are early prefibrilar assemblies but discard the hypothesis that the membrane destabilization is Key event to subsequent disruption of cellular processes, such as ion balance, oxidative state and the eventually cell death

  5. Packaging protein drugs as bacterial inclusion bodies for therapeutic applications

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    Villaverde Antonio

    2012-06-01

    Full Text Available Abstract A growing number of insights on the biology of bacterial inclusion bodies (IBs have revealed intriguing utilities of these protein particles. Since they combine mechanical stability and protein functionality, IBs have been already exploited in biocatalysis and explored for bottom-up topographical modification in tissue engineering. Being fully biocompatible and with tuneable bio-physical properties, IBs are currently emerging as agents for protein delivery into mammalian cells in protein-replacement cell therapies. So far, IBs formed by chaperones (heat shock protein 70, Hsp70, enzymes (catalase and dihydrofolate reductase, grow factors (leukemia inhibitory factor, LIF and structural proteins (the cytoskeleton keratin 14 have been shown to rescue exposed cells from a spectrum of stresses and restore cell functions in absence of cytotoxicity. The natural penetrability of IBs into mammalian cells (reaching both cytoplasm and nucleus empowers them as an unexpected platform for the controlled delivery of essentially any therapeutic polypeptide. Production of protein drugs by biopharma has been traditionally challenged by IB formation. However, a time might have arrived in which recombinant bacteria are to be engineered for the controlled packaging of therapeutic proteins as nanoparticulate materials (nanopills, for their extra- or intra-cellular release in medicine and cosmetics.

  6. Chemical Assistance in Refolding of Bacterial Inclusion Bodies

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    Mona Alibolandi

    2011-01-01

    Full Text Available Escherichia coli is one of the most widely used hosts for the production of recombinant proteins but insoluble expression of heterologous proteins is a major bottleneck in production of recombinant proteins in E. coli. In vitro refolding of inclusion body into proteins with native conformations is a solution for this problem but there is a need for optimization of condition for each protein specifically. Several approaches have been described for in vitro refolding; most of them involve the use of additives for assisting correct folding. Cosolutes play a major role in refolding process and can be classified according to their function as aggregation suppressors and folding enhancers. This paper presents a review of additives that are used in refolding process of insoluble recombinant proteins in small scale and industrial processes.

  7. Using bacterial inclusion bodies to screen for amyloid aggregation inhibitors

    Science.gov (United States)

    2012-01-01

    Background The amyloid-β peptide (Aβ42) is the main component of the inter-neuronal amyloid plaques characteristic of Alzheimer's disease (AD). The mechanism by which Aβ42 and other amyloid peptides assemble into insoluble neurotoxic deposits is still not completely understood and multiple factors have been reported to trigger their formation. In particular, the presence of endogenous metal ions has been linked to the pathogenesis of AD and other neurodegenerative disorders. Results Here we describe a rapid and high-throughput screening method to identify molecules able to modulate amyloid aggregation. The approach exploits the inclusion bodies (IBs) formed by Aβ42 when expressed in bacteria. We have shown previously that these aggregates retain amyloid structural and functional properties. In the present work, we demonstrate that their in vitro refolding is selectively sensitive to the presence of aggregation-promoting metal ions, allowing the detection of inhibitors of metal-promoted amyloid aggregation with potential therapeutic interest. Conclusions Because IBs can be produced at high levels and easily purified, the method overcomes one of the main limitations in screens to detect amyloid modulators: the use of expensive and usually highly insoluble synthetic peptides. PMID:22553999

  8. Tunable geometry of bacterial inclusion bodies as substrate materials for tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    GarcIa-Fruitos, Elena; Seras-Franzoso, JoaquIn; Vazquez, Esther; Villaverde, Antonio [CIBER en BioingenierIa, Biomateriales y Nanomedicina, Bellaterra, 08193 Barcelona (Spain); Institut de Biotecnologia i de Biomedicina and Departament de Genetica i de Microbiologia, Universitat Autonoma de Barcelona, 08193 Bellaterra (Cerdanyola del Valles), Barcelona (Spain)

    2010-05-21

    A spectrum of materials for biomedical applications is produced in bacteria, and some of them, such as metals or polyhydroxyalkanoates, are straightforwardly obtained as particulate entities. We have explored the biofabrication process of bacterial inclusion bodies, particulate proteinaceous materials (ranging from 50 to 500 nm in diameter) recently recognized as suitable for surface topographical modification and tissue engineering. Inclusion bodies have been widely described as spherical or pseudo-spherical particles with only minor morphological variability, mostly restricted to their size. Here we have identified a cellular gene in Escherichia coli (clpP) that controls the in vivo fabrication process of inclusion bodies. In the absence of the encoded protease, the dynamics of protein deposition is perturbed, resulting in unusual tear-shaped particles with enhanced surface-volume ratios. This fact modifies the ability of inclusion bodies to promote mammalian cell attachment and differentiation upon surface decoration. The implications of the genetic control of inclusion body geometry are discussed in the context of their biological fabrication and regarding the biomedical potential of these protein clusters in regenerative medicine.

  9. Tunable geometry of bacterial inclusion bodies as substrate materials for tissue engineering

    Science.gov (United States)

    García-Fruitós, Elena; Seras-Franzoso, Joaquín; Vazquez, Esther; Villaverde, Antonio

    2010-05-01

    A spectrum of materials for biomedical applications is produced in bacteria, and some of them, such as metals or polyhydroxyalkanoates, are straightforwardly obtained as particulate entities. We have explored the biofabrication process of bacterial inclusion bodies, particulate proteinaceous materials (ranging from 50 to 500 nm in diameter) recently recognized as suitable for surface topographical modification and tissue engineering. Inclusion bodies have been widely described as spherical or pseudo-spherical particles with only minor morphological variability, mostly restricted to their size. Here we have identified a cellular gene in Escherichia coli (clpP) that controls the in vivo fabrication process of inclusion bodies. In the absence of the encoded protease, the dynamics of protein deposition is perturbed, resulting in unusual tear-shaped particles with enhanced surface-volume ratios. This fact modifies the ability of inclusion bodies to promote mammalian cell attachment and differentiation upon surface decoration. The implications of the genetic control of inclusion body geometry are discussed in the context of their biological fabrication and regarding the biomedical potential of these protein clusters in regenerative medicine.

  10. Tunable geometry of bacterial inclusion bodies as substrate materials for tissue engineering

    International Nuclear Information System (INIS)

    GarcIa-Fruitos, Elena; Seras-Franzoso, JoaquIn; Vazquez, Esther; Villaverde, Antonio

    2010-01-01

    A spectrum of materials for biomedical applications is produced in bacteria, and some of them, such as metals or polyhydroxyalkanoates, are straightforwardly obtained as particulate entities. We have explored the biofabrication process of bacterial inclusion bodies, particulate proteinaceous materials (ranging from 50 to 500 nm in diameter) recently recognized as suitable for surface topographical modification and tissue engineering. Inclusion bodies have been widely described as spherical or pseudo-spherical particles with only minor morphological variability, mostly restricted to their size. Here we have identified a cellular gene in Escherichia coli (clpP) that controls the in vivo fabrication process of inclusion bodies. In the absence of the encoded protease, the dynamics of protein deposition is perturbed, resulting in unusual tear-shaped particles with enhanced surface-volume ratios. This fact modifies the ability of inclusion bodies to promote mammalian cell attachment and differentiation upon surface decoration. The implications of the genetic control of inclusion body geometry are discussed in the context of their biological fabrication and regarding the biomedical potential of these protein clusters in regenerative medicine.

  11. Strategies for the recovery of active proteins through refolding of bacterial inclusion body proteins

    Directory of Open Access Journals (Sweden)

    Rinas Ursula

    2004-09-01

    Full Text Available Abstract Recent advances in generating active proteins through refolding of bacterial inclusion body proteins are summarized in conjunction with a short overview on inclusion body isolation and solubilization procedures. In particular, the pros and cons of well-established robust refolding techniques such as direct dilution as well as less common ones such as diafiltration or chromatographic processes including size exclusion chromatography, matrix- or affinity-based techniques and hydrophobic interaction chromatography are discussed. Moreover, the effect of physical variables (temperature and pressure as well as the presence of buffer additives on the refolding process is elucidated. In particular, the impact of protein stabilizing or destabilizing low- and high-molecular weight additives as well as micellar and liposomal systems on protein refolding is illustrated. Also, techniques mimicking the principles encountered during in vivo folding such as processes based on natural and artificial chaperones and propeptide-assisted protein refolding are presented. Moreover, the special requirements for the generation of disulfide bonded proteins and the specific problems and solutions, which arise during process integration are discussed. Finally, the different strategies are examined regarding their applicability for large-scale production processes or high-throughput screening procedures.

  12. Bacterial inclusion bodies as potential synthetic devices for pathogen recognition and a therapeutic substance release.

    Science.gov (United States)

    Talafová, Klaudia; Hrabárová, Eva; Chorvát, Dušan; Nahálka, Jozef

    2013-02-07

    Adhesins of pathogens recognise the glycans on the host cell and mediate adherence. They are also crucial for determining the tissue preferences of pathogens. Currently, glyco-nanomaterials provide potential tool for antimicrobial therapy. We demonstrate that properly glyco-tailored inclusion bodies can specifically bind pathogen adhesins and release therapeutic substances. In this paper, we describe the preparation of tailored inclusion bodies via the conjugation of indicator protein aggregated to form inclusion bodies with soluble proteins. Whereas the indicator protein represents a remedy, the soluble proteins play a role in pathogen recognition. For conjugation, glutaraldehyde was used as linker. The treatment of conjugates with polar lysine, which was used to inactivate the residual glutaraldehyde, inhibited unwanted hydrophobic interactions between inclusion bodies. The tailored inclusion bodies specifically interacted with the SabA adhesin from Helicobacter pylori aggregated to form inclusion bodies that were bound to the sialic acids decorating the surface of human erythrocytes. We also tested the release of indicator proteins from the inclusion bodies using sortase A and Ssp DNAB intein self-cleaving modules, respectively. Sortase A released proteins in a relatively short period of time, whereas the intein cleavage took several weeks. The tailored inclusion bodies are promising "nanopills" for biomedical applications. They are able to specifically target the pathogen, while a self-cleaving module releases a soluble remedy. Various self-cleaving modules can be enabled to achieve the diverse pace of remedy release.

  13. L-arginine mediated renaturation enhances yield of human, α6 Type IV collagen non-collagenous domain from bacterial inclusion bodies.

    Science.gov (United States)

    Gunda, Venugopal; Boosani, Chandra Shekhar; Verma, Raj Kumar; Guda, Chittibabu; Sudhakar, Yakkanti Akul

    2012-10-01

    The anti-angiogenic, carboxy terminal non-collagenous domain (NC1) derived from human Collagen type IV alpha 6 chain, [α6(IV)NC1] or hexastatin, was earlier obtained using different recombinant methods of expression in bacterial systems. However, the effect of L-arginine mediated renaturation in enhancing the relative yields of this protein from bacterial inclusion bodies has not been evaluated. In the present study, direct stirring and on-column renaturation methods using L-arginine and different size exclusion chromatography matrices were applied for enhancing the solubility in purifying the recombinant α6(IV)NC1 from bacterial inclusion bodies. This methodology enabled purification of higher quantities of soluble protein from inclusion bodies, which inhibited endothelial cell proliferation, migration and tube formation. Thus, the scope for L-arginine mediated renaturation in obtaining higher yields of soluble, biologically active NC1 domain from bacterial inclusion bodies was evaluated.

  14. Inclusion-Body Myositis: Diagnosis

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    ... for MDA Blog Donate Search MDA.org Close Inclusion-Body Myositis (IBM) Share print email share facebook ... As with other muscle diseases, a doctor diagnoses inclusion-body myositis (IBM) by considering the individual’s personal ...

  15. Thioflavin-S staining of bacterial inclusion bodies for the fast, simple, and inexpensive screening of amyloid aggregation inhibitors.

    Science.gov (United States)

    Pouplana, S; Espargaro, A; Galdeano, C; Viayna, E; Sola, I; Ventura, S; Muñoz-Torrero, D; Sabate, R

    2014-01-01

    Amyloid aggregation is linked to a large number of human disorders, from neurodegenerative diseases as Alzheimer's disease (AD) or spongiform encephalopathies to non-neuropathic localized diseases as type II diabetes and cataracts. Because the formation of insoluble inclusion bodies (IBs) during recombinant protein production in bacteria has been recently shown to share mechanistic features with amyloid self-assembly, bacteria have emerged as a tool to study amyloid aggregation. Herein we present a fast, simple, inexpensive and quantitative method for the screening of potential anti-aggregating drugs. This method is based on monitoring the changes in the binding of thioflavin-S to intracellular IBs in intact Eschericchia coli cells in the presence of small chemical compounds. This in vivo technique fairly recapitulates previous in vitro data. Here we mainly use the Alzheimer's related β-amyloid peptide as a model system, but the technique can be easily implemented for screening inhibitors relevant for other conformational diseases simply by changing the recombinant amyloid protein target. Indeed, we show that this methodology can be also applied to the evaluation of inhibitors of the aggregation of tau protein, another amyloidogenic protein with a key role in AD.

  16. Inclusion body myositis. Clinical aspects

    NARCIS (Netherlands)

    Cox, Fieke Maria Elisabeth

    2014-01-01

    Sporadische inclusion body myositis (IBM) is een van de meest voor voorkomende verworven spierziekte die ontstaat na het 50e levensjaar. In dit proefschrift worden de klinische aspecten van sporadische IBM beschreven. Uit de studie met betrekking tot het natuurlijk beloop blijkt dat de ziekte niet

  17. One-step affinity tag purification of full-length recombinant human AP-1 complexes from bacterial inclusion bodies using a polycistronic expression system.

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    Wang, Wei-Ming; Lee, A-Young; Chiang, Cheng-Ming

    2008-05-01

    The AP-1 transcription factor is a dimeric protein complex formed primarily between Jun (c-Jun, JunB, JunD) and Fos (c-Fos, FosB, Fra-1, Fra-2) family members. These distinct AP-1 complexes are expressed in many cell types and modulate target gene expression implicated in cell proliferation, differentiation, and stress responses. Although the importance of AP-1 has long been recognized, the biochemical characterization of AP-1 remains limited in part due to the difficulty in purifying full-length, reconstituted dimers with active DNA-binding and transcriptional activity. Using a combination of bacterial coexpression and epitope-tagging methods, we successfully purified all 12 heterodimers (3 Junx4 Fos) of full-length human AP-1 complexes as well as c-Jun/c-Jun, JunD/JunD, and c-Jun/JunD dimers from bacterial inclusion bodies using one-step nickel-NTA affinity tag purification following denaturation and renaturation of coexpressed AP-1 subunits. Coexpression of two constitutive components in a dimeric AP-1 complex helps stabilize the proteins when compared with individual protein expression in bacteria. Purified dimeric AP-1 complexes are functional in sequence-specific DNA binding, as illustrated by electrophoretic mobility shift assays and DNase I footprinting, and are also active in transcription with in vitro-reconstituted human papillomavirus (HPV) chromatin containing AP-1-binding sites in the native configuration of HPV nucleosomes. The availability of these recombinant full-length human AP-1 complexes has greatly facilitated mechanistic studies of AP-1-regulated gene transcription in many biological systems.

  18. Inclusion body myositis : a nationwide study

    NARCIS (Netherlands)

    Badrising, Umesh Arvind

    2006-01-01

    The studies reported in this thesis aimed to survey the prevalence of inclusion body myositis (IBM), to describe its clinical features and course, to investigate whether the major histocompatibility complex predisposed subjects to IBM and autoimmune disorders (AID), to investigate the possible

  19. A study of the cytoplasmic expression of a form of human prolactin and of its solubilization and renaturation from bacterial inclusion bodies

    International Nuclear Information System (INIS)

    Affonso, Regina

    2000-01-01

    Different vector elements, that can determine a high expression level of a form of human prolactin (taghPrl) in bacterial cytoplasm, were studied. Expression conditions were first optimized for a reference vector, which was used to transform different strains of E. coli: HB2151, RRI and RB791. The highest expression level (113 ±16 μg/mL.A 600 ) was obtained in HB2151, after activation with only 0.1 mM IPTG. At this point the influence of the transcription terminator (g32 from bacteriophage T4), of the translation enhancer (g10 from bacteriophage T7), of the promoter (λP L or tac) and of the antibiotic resistance gene (amp r or kan r ) were studied. The first three elements did not show any significant influence, at least in our systems. On the contrary, the analysis of the influence of amp r and kan r genes showed, unexpectedly, that the presence of the last one provides an approximately 5-fold higher expression for taghPrl in E. coli cytoplasm. Finally, an appropriate extraction, solubilization, renaturation and purification process, able to provide a monomeric form of taghPrl, was studied. A method utilizing urea and mercaptoethanol as solubilizing agents and a dialysis as a renaturation procedure, provided with some modifications, one of the highest yields ever reported in the literature: 35.4 ± 4.5% of total recovery. Moreover, the biological activity of the taghPrl obtained, when tested in the Nb2 cell proliferation assay, was of the same order of that shown by the International Standard of human prolactin of pituitary origin. These data show that the cytoplasmic expression system here described, which can provide an expression efficiency 50-100 - fold higher than the periplasmic expression, can represent a valid alternative for the production of this and of other hormones of pharmaceutical interest and grade. (author)

  20. Genetics Home Reference: familial encephalopathy with neuroserpin inclusion bodies

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    ... Home Health Conditions FENIB Familial encephalopathy with neuroserpin inclusion bodies Printable PDF Open All Close All Enable ... expand/collapse boxes. Description Familial encephalopathy with neuroserpin inclusion bodies ( FENIB ) is a disorder that causes progressive ...

  1. Dynamic recruitment of active proteasomes into polyglutamine initiated inclusion bodies

    NARCIS (Netherlands)

    Schipper-Krom, Sabine; Juenemann, Katrin; Jansen, Anne H.; Wiemhoefer, Anne; van den Nieuwendijk, Rianne; Smith, Donna L.; Hink, Mark A.; Bates, Gillian P.; Overkleeft, Hermen; Ovaa, Huib; Reits, Eric

    2014-01-01

    Neurodegenerative disorders such as Huntington's disease are hallmarked by neuronal intracellular inclusion body formation. Whether proteasomes are irreversibly recruited into inclusion bodies in these protein misfolding disorders is a controversial subject. In addition, it has been proposed that

  2. Renal pathophysiologic role of cortical tubular inclusion bodies.

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    Radi, Zaher A; Stewart, Zachary S; Grzemski, Felicity A; Bobrowski, Walter F

    2013-01-01

    Renal tubular inclusion bodies are rarely associated with drug administration. The authors describe the finding of renal cortical tubular intranuclear and intracytoplasmic inclusion bodies associated with the oral administration of a norepinephrine/serotonin reuptake inhibitor (NSRI) test article in Sprague-Dawley (SD) rats. Rats were given an NSRI daily for 4 weeks, and kidney histopathologic, ultrastructural pathology, and immunohistochemical examinations were performed. Round eosinophilic intranuclear inclusion bodies were observed histologically in the tubular epithelial cells of the renal cortex in male and female SD rats given the NSRI compound. No evidence of degeneration or necrosis was noted in the inclusion-containing renal cells. By ultrastructural pathology, inclusion bodies consisted of finely granular, amorphous, and uniformly stained nonmembrane-bound material. By immunohistochemistry, inclusion bodies stained positive for d-amino acid oxidase (DAO) protein. In addition, similar inclusion bodies were noted in the cytoplasmic tubular epithelial compartment by ultrastructural and immunohistochemical examination.  This is the first description of these renal inclusion bodies after an NSRI test article administration in SD rats. Such drug-induced renal inclusion bodies are rat-specific, do not represent an expression of nephrotoxicity, represent altered metabolism of d-amino acids, and are not relevant to human safety risk assessment.

  3. Efficient renaturation of inclusion body proteins denatured by SDS.

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    He, Chuan; Ohnishi, Kouhei

    2017-09-02

    Inclusion bodies are often formed when the foreign protein is over expressed in Escherichia coli. Since proteins in inclusion bodies are inactive, denaturing and refolding of inclusion body proteins are necessary to obtain the active form. Instead of the conventional denaturants, urea and guanidine hydrochloride, a strong anionic detergent SDS was used to solubilize C-terminal His-tag form of ulvan lyase in the inclusion bodies. Solution containing SDS-solubilized enzyme were kept on ice to precipitate SDS, followed by SDS-KCl insoluble crystal formation to remove SDS completely. After removing the precipitate by centrifugation, the supernatant was applied to Ni-NTA column to purify His-tagged ulvan lyase. The purified protein showed a dimeric form and ulvan lyase activity, demonstrating that SDS-denatured protein was renatured and recovered enzyme activity. This simple method could be useful for refolding other inclusion body proteins. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Inclusion bodies are a site of ebolavirus replication.

    Science.gov (United States)

    Hoenen, Thomas; Shabman, Reed S; Groseth, Allison; Herwig, Astrid; Weber, Michaela; Schudt, Gordian; Dolnik, Olga; Basler, Christopher F; Becker, Stephan; Feldmann, Heinz

    2012-11-01

    Inclusion bodies are a characteristic feature of ebolavirus infections in cells. They contain large numbers of preformed nucleocapsids, but their biological significance has been debated, and they have been suggested to be aggregates of viral proteins without any further biological function. However, recent data for other viruses that produce similar structures have suggested that inclusion bodies might be involved in genome replication and transcription. In order to study filovirus inclusion bodies, we fused mCherry to the ebolavirus polymerase L, which is found in inclusion bodies. The resulting L-mCherry fusion protein was functional in minigenome assays and incorporated into virus-like particles. Importantly, L-mCherry fluorescence in transfected cells was readily detectable and distributed in a punctate pattern characteristic for inclusion bodies. A recombinant ebolavirus encoding L-mCherry instead of L was rescued and showed virtually identical growth kinetics and endpoint titers to those for wild-type virus. Using this virus, we showed that the onset of inclusion body formation corresponds to the onset of viral genome replication, but that viral transcription occurs prior to inclusion body formation. Live-cell imaging further showed that inclusion bodies are highly dynamic structures and that they can undergo dramatic reorganization during cell division. Finally, by labeling nascent RNAs using click technology we showed that inclusion bodies are indeed the site of viral RNA synthesis. Based on these data we conclude that, rather than being inert aggregates of nucleocapsids, ebolavirus inclusion bodies are in fact complex and dynamic structures and an important site at which viral RNA replication takes place.

  5. Parameter identification problems for thin inclusions in elastic bodies

    Science.gov (United States)

    Khludnev, A. M.; Popova, T. S.

    2017-10-01

    The paper concerns an identification of rigidity parameters for thin inclusions located inside elastic bodies. A delamination of the inclusions is assumed thus providing a crack between inclusions and the elastic matrix. Inequality type boundary conditions are imposed at the crack faces to exclude a mutual penetration. We consider elastic as well as rigid inclusions and solve an optimal control problem for finding a rigidity parameter minimizing a suitable cost functional. The cost functional characterizes a displacement of the inclusion, and a rigidity parameter serves as a control function. We prove a solution existence of the problems formulated.

  6. Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process.

    Science.gov (United States)

    Singh, Anupam; Upadhyay, Vaibhav; Upadhyay, Arun Kumar; Singh, Surinder Mohan; Panda, Amulya Kumar

    2015-03-25

    Formation of inclusion bodies in bacterial hosts poses a major challenge for large scale recovery of bioactive proteins. The process of obtaining bioactive protein from inclusion bodies is labor intensive and the yields of recombinant protein are often low. Here we review the developments in the field that are targeted at improving the yield, as well as quality of the recombinant protein by optimizing the individual steps of the process, especially solubilization of the inclusion bodies and refolding of the solubilized protein. Mild solubilization methods have been discussed which are based on the understanding of the fact that protein molecules in inclusion body aggregates have native-like structure. These methods solubilize the inclusion body aggregates while preserving the native-like protein structure. Subsequent protein refolding and purification results in high recovery of bioactive protein. Other parameters which influence the overall recovery of bioactive protein from inclusion bodies have also been discussed. A schematic model describing the utility of mild solubilization methods for high throughput recovery of bioactive protein has also been presented.

  7. Dynamic recruitment of ubiquitin to mutant huntingtin inclusion bodies

    NARCIS (Netherlands)

    Juenemann, Katrin; Jansen, Anne H. P.; van Riel, Luigi; Merkx, Remco; Mulder, Monique P. C.; An, Heeseon; Statsyuk, Alexander; Kirstein, Janine; Ovaa, Huib; Reits, Eric A.

    2018-01-01

    Many neurodegenerative diseases, such as Huntington's disease, are hallmarked by the formation of intracellular inclusion bodies (IBs) that are decorated with ubiquitin, proteasomes and chaperones. The apparent enrichment of ubiquitin and components involved in protein quality control at IBs

  8. Yeast prions form infectious amyloid inclusion bodies in bacteria

    Directory of Open Access Journals (Sweden)

    Espargaró Alba

    2012-06-01

    Full Text Available Abstract Background Prions were first identified as infectious proteins associated with fatal brain diseases in mammals. However, fungal prions behave as epigenetic regulators that can alter a range of cellular processes. These proteins propagate as self-perpetuating amyloid aggregates being an example of structural inheritance. The best-characterized examples are the Sup35 and Ure2 yeast proteins, corresponding to [PSI+] and [URE3] phenotypes, respectively. Results Here we show that both the prion domain of Sup35 (Sup35-NM and the Ure2 protein (Ure2p form inclusion bodies (IBs displaying amyloid-like properties when expressed in bacteria. These intracellular aggregates template the conformational change and promote the aggregation of homologous, but not heterologous, soluble prionogenic molecules. Moreover, in the case of Sup35-NM, purified IBs are able to induce different [PSI+] phenotypes in yeast, indicating that at least a fraction of the protein embedded in these deposits adopts an infectious prion fold. Conclusions An important feature of prion inheritance is the existence of strains, which are phenotypic variants encoded by different conformations of the same polypeptide. We show here that the proportion of infected yeast cells displaying strong and weak [PSI+] phenotypes depends on the conditions under which the prionogenic aggregates are formed in E. coli, suggesting that bacterial systems might become useful tools to generate prion strain diversity.

  9. Dynamic recruitment of active proteasomes into polyglutamine initiated inclusion bodies.

    Science.gov (United States)

    Schipper-Krom, Sabine; Juenemann, Katrin; Jansen, Anne H; Wiemhoefer, Anne; van den Nieuwendijk, Rianne; Smith, Donna L; Hink, Mark A; Bates, Gillian P; Overkleeft, Hermen; Ovaa, Huib; Reits, Eric

    2014-01-03

    Neurodegenerative disorders such as Huntington's disease are hallmarked by neuronal intracellular inclusion body formation. Whether proteasomes are irreversibly recruited into inclusion bodies in these protein misfolding disorders is a controversial subject. In addition, it has been proposed that the proteasomes may become clogged by the aggregated protein fragments, leading to impairment of the ubiquitin-proteasome system. Here, we show by fluorescence pulse-chase experiments in living cells that proteasomes are dynamically and reversibly recruited into inclusion bodies. As these recruited proteasomes remain catalytically active and accessible to substrates, our results challenge the concept of proteasome sequestration and impairment in Huntington's disease, and support the reported absence of proteasome impairment in mouse models of Huntington's disease. Copyright © 2013 Federation of European Biochemical Societies. All rights reserved.

  10. Inclusion bodies in loggerhead erythrocytes are associated with unstable hemoglobin and resemble human Heinz bodies.

    Science.gov (United States)

    Basile, Filomena; Di Santi, Annalisa; Caldora, Mercedes; Ferretti, Luigi; Bentivegna, Flegra; Pica, Alessandra

    2011-08-01

    The aim of this study was to clarify the role of the erythrocyte inclusions found during the hematological screening of loggerhead population of the Mediterranean Sea. We studied the erythrocyte inclusions in blood specimens collected from six juvenile and nine adult specimens of the loggerhead turtle, Caretta caretta, from the Adriatic and Tyrrhenian Seas. Our study indicates that the percentage of mature erythrocytes containing inclusions ranged from 3 to 82%. Each erythrocyte contained only one round inclusion body. Inclusion bodies stained with May Grünwald-Giemsa show that their cytochemical and ultrastructure characteristics are identical to those of human Heinz bodies. Because Heinz bodies originate from the precipitation of unstable hemoglobin (Hb) and cause globular osmotic resistance to increase, we analyzed loggerhead Hb using electrophoresis and high-performance liquid chromatography to detect and quantitate Hb fractions. We also tested the resistance of Hb to alkaline pH, heat, isopropanol denaturation, and globular osmosis. Our hemogram results excluded the occurrence of any infection, which could be associated with an inclusion body, in all the specimens. Negative Feulgen staining indicated that the inclusion bodies are not derived from DNA fragmentation. We hypothesize that amino acid substitutions could explain why loggerhead Hb precipitates under normal physiologic conditions, forming Heinz bodies. The identification of inclusion bodies in loggerhead erythrocytes allow us to better understand the haematological characteristics and the physiology of these ancient reptiles, thus aiding efforts to conserve such an endangered species. Copyright © 2011 Wiley-Liss, Inc., A Wiley Company.

  11. [Solubilization Specificities Interferon beta-1b from Inclusion Bodies].

    Science.gov (United States)

    Zhuravko, A S; Kononova, N V; Bobruskin, A I

    2015-01-01

    A new solubilization method of recombinant interferon beta-1b (IFNβ-1b) from the inclusion bodies was developed. This method allows to extract the target protein selectively in the solutions of different alcohols, such as ethanol, propanol and isopropanol. It was shown that the more effective IFNβ-1b solubilization was achieved in the 55% propanol solution. This method allowed to extract the target protein from inclusion bodies around 85-90%, and significantly reduced Escherichia coli content in the solubilizate, in comparison with standard methods.

  12. [RBCs inclusions after splenectomy: not only Howell-Jolly bodies!].

    Science.gov (United States)

    Solis, Morgane; Perrin, Julien; Guédenet, Jean-Claude; Lesesve, Jean-François

    2013-01-01

    We report on several abnormalities of the red blood cells, in particular erythrocytic inclusions, in a 49 year old man. The blood smear presented with marked anisocytosis, target cells, acanthocytes and Howell-Jolly bodies, as well as Pappenheimer bodies and basophilic stippling. These abnormalities can be explained by the clinical context which associated splenectomy, iron overload and severe denutrition. The patient died a few weeks after from kidney and liver insufficiency. We comment on the different features of erythrocytic inclusions and the clinical situations in which they can be found.

  13. Lewy and his inclusion bodies: Discovery and rejection

    Directory of Open Access Journals (Sweden)

    Eliasz Engelhardt

    Full Text Available ABSTRACT Fritz Jacob Heinrich Lewy described the pathology of Paralysis agitans [Parkinson disease] and was the first to identify eosinophilic inclusion bodies in neurons of certain brain nuclei, later known as Lewy bodies, the pathological signature of the Lewy body diseases. In 1912, he published his seminal study, followed soon after by an update paper, and 10 years later, in 1923, by his voluminous book, where he exhaustively described the subject. The publication provided extensive information on the pathology of Paralysis agitans, and the entirely novel finding of eosinophilic inclusion bodies, which would become widely recognized and debated in the future. His discovery was acknowledged by important researchers who even named the structure after him. However, after his last publication on the issue, inexplicably, he never mentioned his histopathological discovery again. Despite several hypotheses, the reasons that led him to neglect (reject the structure which he so preeminently described have remained elusive.

  14. [Inclusion Bodies are Formed in SFTSV-infected Human Macrophages].

    Science.gov (United States)

    Jin, Cong; Song, Jingdong; Han, Ying; Li, Chuan; Qiu, Peihong; Liang, Mifang

    2016-01-01

    The severe fever with thrombocytopenia syndrome virus (SFTSV) is a new member in the genus Phlebovirus of the family Bunyaviridae identified in China. The SFTSV is also the causative pathogen of an emerging infectious disease: severe fever with thrombocytopenia syndrome. Using immunofluorescent staining and confocal microscopy, the intracellular distribution of nucleocapsid protein (NP) in SFTSV-infected THP-1 cells was investigated with serial doses of SFTSV at different times after infection. Transmission electron microscopy was used to observe the ultrafine intracellular structure of SFTSV-infected THP-1 cells at different times after infection. SFTSV NP could form intracellular inclusion bodies in infected THP-1 cells. The association between NP-formed inclusion bodies and virus production was analyzed: the size of the inclusion body formed 3 days after infection was correlated with the viral load in supernatants collected 7 days after infection. These findings suggest that the inclusion bodies formed in SFTSV-infected THP-1 cells could be where the SFTSV uses host-cell proteins and intracellular organelles to produce new viral particles.

  15. Richtlijn 'dermatomyositis, polymyositis en sporadische "inclusion body"-myositis'

    NARCIS (Netherlands)

    Hoogendijk, J. E.; Bijlsma, J. W. J.; van Engelen, B. G. M.; Lindeman, E.; van Royen-Kerkhof, A.; de Rie, M. A.; de Visser, M.; Jennekens, F. G. I.

    2005-01-01

    This guideline presents recommendations for the diagnosis and treatment of dermatomyositis, polymyositis and sporadic inclusion body myositis (sIBM) according to the best available evidence. Characteristic skin abnormalities can be sufficient for the diagnosis of dermatomyositis. In case of doubt, a

  16. [The practice guideline 'Dermatomyositis, polymyositis and sporadic inclusion body myositis'

    NARCIS (Netherlands)

    Hoogendijk, J.E.; Bijlsma, J.W.J.; Engelen, B.G.M. van; Lindeman, E.J.M.; Royen-Kerkhof, A. van; Rie, M.A. de; Visser, M. de; Jennekens, F.G.I.

    2005-01-01

    This guideline presents recommendations for the diagnosis and treatment of dermatomyositis, polymyositis and sporadic inclusion body myositis (sIBM) according to the best available evidence. Characteristic skin abnormalities can be sufficient for the diagnosis of dermatomyositis. In case of doubt, a

  17. High throughput inclusion body sizing: Nano particle tracking analysis.

    Science.gov (United States)

    Reichelt, Wieland N; Kaineder, Andreas; Brillmann, Markus; Neutsch, Lukas; Taschauer, Alexander; Lohninger, Hans; Herwig, Christoph

    2017-06-01

    The expression of pharmaceutical relevant proteins in Escherichia coli frequently triggers inclusion body (IB) formation caused by protein aggregation. In the scientific literature, substantial effort has been devoted to the quantification of IB size. However, particle-based methods used up to this point to analyze the physical properties of representative numbers of IBs lack sensitivity and/or orthogonal verification. Using high pressure freezing and automated freeze substitution for transmission electron microscopy (TEM) the cytosolic inclusion body structure was preserved within the cells. TEM imaging in combination with manual grey scale image segmentation allowed the quantification of relative areas covered by the inclusion body within the cytosol. As a high throughput method nano particle tracking analysis (NTA) enables one to derive the diameter of inclusion bodies in cell homogenate based on a measurement of the Brownian motion. The NTA analysis of fixated (glutaraldehyde) and non-fixated IBs suggests that high pressure homogenization annihilates the native physiological shape of IBs. Nevertheless, the ratio of particle counts of non-fixated and fixated samples could potentially serve as factor for particle stickiness. In this contribution, we establish image segmentation of TEM pictures as an orthogonal method to size biologic particles in the cytosol of cells. More importantly, NTA has been established as a particle-based, fast and high throughput method (1000-3000 particles), thus constituting a much more accurate and representative analysis than currently available methods. Copyright © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Role of the disaggregase ClpB in processing of proteins aggregated as inclusion bodies.

    Science.gov (United States)

    Zblewska, Kamila; Krajewska, Joanna; Zolkiewski, Michal; Kędzierska-Mieszkowska, Sabina

    2014-08-01

    Overproduction of heterologous proteins in bacterial systems often results in the formation of insoluble inclusion bodies (IBs), which is a major impediment in biochemical research and biotechnology. In principle, the activity of molecular chaperones could be employed to gain control over the IB formation and to improve the recombinant protein yields, but the potential of each of the major bacterial chaperones (DnaK/J, GroEL/ES, and ClpB) to process IBs has not been fully established yet. We investigated the formation of inclusion bodies (IBs) of two aggregation-prone proteins, VP1LAC and VP1GFP, overproduced in Escherichiacoli in the presence and absence of the chaperone ClpB. We found that both ClpB isoforms, ClpB95 and ClpB80 accumulated in E. coli cells during the production of IBs. The amount of IB proteins increased in the absence of ClpB. ClpB supported the resolubilization and reactivation of the aggregated VP1LAC and VP1GFP in E. coli cells. The IB disaggregation was optimal in the presence of both ClpB95 and ClpB80. Our results indicate an essential role of ClpB in controlling protein aggregation and inclusion body formation in bacteria. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Soni-removal of nucleic acids from inclusion bodies.

    Science.gov (United States)

    Neerathilingam, Muniasamy; Mysore, Sumukh; Gandham, Sai Hari A

    2014-05-23

    Inclusion bodies (IBs) are commonly formed in Escherichia coli due to over expression of recombinant proteins in non-native state. Isolation, denaturation and refolding of these IBs is generally performed to obtain functional protein. However, during this process IBs tend to form non-specific interactions with sheared nucleic acids from the genome, thus getting carried over into downstream processes. This may hinder the refolding of IBs into their native state. To circumvent this, we demonstrate a methodology termed soni-removal which involves disruption of nucleic acid-inclusion body interaction using sonication; followed by solvent based separation. As opposed to conventional techniques that use enzymes and column-based separations, soni-removal is a cost effective alternative for complete elimination of buried and/or strongly bound short nucleic acid contaminants from IBs. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  20. Inclusion body myositis: diagnosis, pathogenesis, and treatment options.

    Science.gov (United States)

    Solorzano, Guillermo E; Phillips, Lawrence H

    2011-05-01

    Inclusion body myositis (IBM) is the most common acquired myopathy in people older than 50 years. IBM typically presents with distal upper extremity weakness accompanied by proximal lower extremity muscle weakness. Associated clinical findings include asymmetric weakness, foot drop, and dysphagia. The pathogenesis of IBM is not clear. In this article the authors briefly discuss postulated pathogenic mechanisms. Although no proven pharmacotherapy exists, some promising candidates are discussed. Copyright © 2011 Elsevier Inc. All rights reserved.

  1. [In vitro renaturation of proteins from inclusion bodies].

    Science.gov (United States)

    Porowińska, Dorota; Marszałek, Ewelina; Wardęcka, Paulina; Komoszyński, Michał

    2012-06-11

    Recombinant proteins and enzymes are commonly used in many areas of our life, such as diagnostics, industry and medicine, due to heterologous synthesis in prokaryotic expression systems. However, a high expression level of foreign protein in bacteria cells results in formation of inactive and insoluble aggregates--inclusion bodies. Reactivation of aggregated proteins is a complex and time-consuming process. Every protein requires experimental optimization of the process conditions. The choice of the refolding method depends on the type of recombinant protein and its physical, chemical and biological properties. Recovery of the activity of proteins accumulated in inclusion bodies can be divided into 4 steps: 1) inclusion bodies isolation, 2) solubilization of aggregates, 3) renaturation, 4) purification of catalytically active molecules. Efficiency of the refolding process depends on many physical factors and chemical and biological agents. The above parameters determine the time of the folding and prevent protein aggregation. They also assist the folding and have an influence on the solubility and stability of native molecules. To date, dilution, dialysis and chromatography are the most often used methods for protein refolding.

  2. Dark proteins: effect of inclusion body formation on quantification of protein expression.

    Science.gov (United States)

    Iafolla, Marco A J; Mazumder, Mostafizur; Sardana, Vandit; Velauthapillai, Tharsan; Pannu, Karanbir; McMillen, David R

    2008-09-01

    Plasmid-borne gene expression systems have found wide application in the emerging fields of systems biology and synthetic biology, where plasmids are used to implement simple network architectures, either to test systems biology hypotheses about issues such as gene expression noise or as a means of exerting artificial control over a cell's dynamics. In both these cases, fluorescent proteins are commonly applied as a means of monitoring the expression of genes in the living cell, and efforts have been made to quantify protein expression levels through fluorescence intensity calibration and by monitoring the partitioning of proteins among the two daughter cells after division; such quantification is important in formulating the predictive models desired in systems and synthetic biology research. A potential pitfall of using plasmid-based gene expression systems is that the high protein levels associated with expression from plasmids can lead to the formation of inclusion bodies, insoluble aggregates of misfolded, nonfunctional proteins that will not generate fluorescence output; proteins caught in these inclusion bodies are thus "dark" to fluorescence-based detection methods. If significant numbers of proteins are incorporated into inclusion bodies rather than becoming biologically active, quantitative results obtained by fluorescent measurements will be skewed; we investigate this phenomenon here. We have created two plasmid constructs with differing average copy numbers, both incorporating an unregulated promoter (P(LtetO-1) in the absence of TetR) expressing the GFP derivative enhanced green fluorescent protein (EGFP), and inserted them into Escherichia coli bacterial cells (a common model organism for work on the dynamics of prokaryotic gene expression). We extracted the inclusion bodies, denatured them, and refolded them to render them active, obtaining a measurement of the average number of EGFP per cell locked into these aggregates; at the same time, we used

  3. Enhanced expression and purification of camelid single domain VHH antibodies from classical inclusion bodies.

    Science.gov (United States)

    Maggi, Maristella; Scotti, Claudia

    2017-08-01

    Single domain antibodies (sdAbs) are small antigen-binding domains derived from naturally occurring, heavy chain-only immunoglobulins isolated from camelid and sharks. They maintain the same binding capability of full-length IgGs but with improved thermal stability and permeability, which justifies their scientific, medical and industrial interest. Several described recombinant forms of sdAbs have been produced in different hosts and with different strategies. Here we present an optimized method for a time-saving, high yield production and extraction of a poly-histidine-tagged sdAb from Escherichia coli classical inclusion bodies. Protein expression and extraction were attempted using 4 different methods (e.g. autoinducing or IPTG-induced soluble expression, non-classical and classical inclusion bodies). The best method resulted to be expression in classical inclusion bodies and urea-mediated protein extraction which yielded 60-70 mg/l bacterial culture. The method we here describe can be of general interest for an enhanced and efficient heterologous expression of sdAbs for research and industrial purposes. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Bacterial communities in the fruit bodies of ground basidiomycetes

    Science.gov (United States)

    Zagryadskaya, Yu. A.; Lysak, L. V.; Chernov, I. Yu.

    2015-06-01

    Fruit bodies of basidiomycetes at different stages of decomposition serve as specific habitats in forest biocenoses for bacteria and differ significantly with respect to the total bacterial population and abundance of particular bacterial genera. A significant increase in the total bacterial population estimated by the direct microscopic method with acridine orange staining and in the population of saprotrophic bacteria (inoculation of glucose peptone yeast agar) in fruit bodies of basidiomycetes Armillaria mellea and Coprinus comatus was recorded at the final stage of their decomposition in comparison with the initial stage. Gramnegative bacteria predominated in the tissues of fruit bodies at all the stages of decomposition and were represented at the final stage by the Aeromonas, Vibrio, and Pseudomonas genera (for fruit bodies of A. mellea) the Pseudomonas genus (for fruit bodies of C. comatus). The potential influence of bacterial communities in the fruit bodies of soil basidiomycetes on the formation of bacterial communities in the upper soil horizons in forest biocenoses is discussed. The loci connected with the development and decomposition of fruit bodies of basidiomycetes on the soil surface are promising for targeted search of Gram-negative bacteria, the important objects of biotechnology.

  5. Formation of distinct inclusion bodies by inhibition of ubiquitin-proteasome and autophagy-lysosome pathways

    International Nuclear Information System (INIS)

    Lee, Junho; Yang, Kyu-Hwan; Joe, Cheol O.; Kang, Seok-Seong

    2011-01-01

    Research highlights: → Distinct inclusion bodies are developed by inhibition of UPP and ALP. → The inclusion bodies differ in morphology, localization and formation process. → The inclusion bodies are distinguishable by the localization of TSC2. → Inhibition of both UPP and ALP simultaneously induces those inclusion bodies. -- Abstract: Accumulation of misfolded proteins is caused by the impairment of protein quality control systems, such as ubiquitin-proteasome pathway (UPP) and autophagy-lysosome pathway (ALP). In this study, the formation of inclusion bodies was examined after the blockade of UPP and/or ALP in A549 cells. UPP inhibition induced a single and large inclusion body localized in microtubule-organizing center. Interestingly, however, ALP inhibition generated dispersed small inclusion bodies in the cytoplasm. Tuberous sclerosis complex 2 was selectively accumulated in the inclusion bodies of UPP-inhibited cells, but not those of ALP-inhibited cells. Blockade of transcription and translation entirely inhibited the formation of inclusion body induced by UPP inhibition, but partially by ALP inhibition. Moreover, the simultaneous inhibition of two protein catabolic pathways independently developed two distinct inclusion bodies within a single cell. These findings clearly demonstrated that dysfunction of each catabolic pathway induced formation and accumulation of unique inclusion bodies on the basis of morphology, localization and formation process in A549 cells.

  6. Interconnected cavernous structure of bacterial fruiting bodies.

    Directory of Open Access Journals (Sweden)

    Cameron W Harvey

    Full Text Available The formation of spore-filled fruiting bodies by myxobacteria is a fascinating case of multicellular self-organization by bacteria. The organization of Myxococcus xanthus into fruiting bodies has long been studied not only as an important example of collective motion of bacteria, but also as a simplified model for developmental morphogenesis. Sporulation within the nascent fruiting body requires signaling between moving cells in order that the rod-shaped self-propelled cells differentiate into spores at the appropriate time. Probing the three-dimensional structure of myxobacteria fruiting bodies has previously presented a challenge due to limitations of different imaging methods. A new technique using Infrared Optical Coherence Tomography (OCT revealed previously unknown details of the internal structure of M. xanthus fruiting bodies consisting of interconnected pockets of relative high and low spore density regions. To make sense of the experimentally observed structure, modeling and computer simulations were used to test a hypothesized mechanism that could produce high-density pockets of spores. The mechanism consists of self-propelled cells aligning with each other and signaling by end-to-end contact to coordinate the process of differentiation resulting in a pattern of clusters observed in the experiment. The integration of novel OCT experimental techniques with computational simulations can provide new insight into the mechanisms that can give rise to the pattern formation seen in other biological systems such as dictyostelids, social amoeba known to form multicellular aggregates observed as slugs under starvation conditions.

  7. Inclusion-body myositis presenting with facial diplegia.

    Science.gov (United States)

    Ghosh, Partha S; Laughlin, Ruple S; Engel, Andrew G

    2014-02-01

    The hallmark clinical presentation of inclusion-body myositis (IBM) is slowly progressive weakness that characteristically affects the quadriceps and finger and wrist finger flexor muscles. Facial weakness can also occur, but it is typically mild and not a prominent finding. We describe the clinical features, laboratory investigations, and muscle biopsy findings in a 58-year old man who presented with a 6-year history of marked progressive symmetrical facial weakness. Examination also showed shoulder abduction and hip extensor weakness. The patient's serum creatine kinase level was 655 U/L, and electromyography showed fibrillation potentials and myopathic motor unit potentials. A biopsy specimen of the left biceps muscle was pathognomonic for IBM. This patient did not have a typical presentation for IBM but rather fulfilled the pathological criteria for IBM. To our knowledge, facial diplegia has not been reported previously as a presenting manifestation of IBM. Copyright © 2013 Wiley Periodicals, Inc.

  8. Physical function and muscle strength in sporadic inclusion body myositis

    DEFF Research Database (Denmark)

    Jørgensen, Anders N; Aagaard, Per; Nielsen, Jakob L

    2017-01-01

    INTRODUCTION: In this study, self-reported physical function, functional capacity, and isolated muscle function were investigated in sporadic inclusion body myositis (sIBM) patients. METHODS: The 36-item Short Form (SF-36) Health Survey and 2-min walk test (2MWT), timed up & go test (TUG), and 30-s...... chair stand performance were evaluated. In addition, patients were tested for knee extensor muscle strength (isokinetic dynamometer) and leg extension power (Nottingham power rig). RESULTS: TUG performance was the strongest predictor of self-reported physical function (r(2) = 0.56, P ... to sensitively predict self-perceived physical function in sIBM patients. Notably, between-limb asymmetry in lower limb muscle strength had a substantial negative impact on motor tasks involving gait function. Muscle Nerve, 2017....

  9. Inclusion-body myositis: clinical, diagnostic, and pathologic aspects.

    Science.gov (United States)

    Engel, W King; Askanas, Valerie

    2006-01-24

    The diagnostic aspects of sporadic inclusion-body myositis (s-IBM), and a few comments on our own approach to its treatment, are presented to foster the goals of this symposium, which was organized to provoke new ideas concerning the cause and treatment of this currently unsolvable disease. s-IBM is the most common, progressive, debilitating muscle disease beginning in persons over age 50 years, and it is more common in men. Diagnostic parameters reviewed are clinical, muscle-biopsy histochemistry, electrophysiologic and CSF evaluations. Overall, the degenerative phenomena in s-IBM muscle fibers seem to be the major cause of the progressive, unstoppable weakness, rather than the lymphocytic inflammation. Available treatments are of only slight, temporary benefit for only some s-IBM patients, indicating a desperate need for definitive therapies.

  10. Seborrheic inclusion cyst of the skin positive for cytoplasmic inclusion bodies and HPV antigen.

    Science.gov (United States)

    Terada, Tadashi

    2012-01-01

    Seborrheic inclusion cyst (SIC) is a very rare variant of epidermal cyst of the skin. SIC shows seborrheic keratosis (SK)-like lesion in epidermal cyst. SIC is extremely rare; only 6 case reports have been published in the English literature. However, no immunohistochemical study of SIC has been reported. A 41-year-old Japanese man noticed a subcutaneous tumor in the neck. Physical examination showed slightly mobile tumor in the subcutaneous tissue, and total excision was performed. Grossly, the tumor (1 x 1 x 0.8 cm) was cyst containing atheromatous keratin. Microscopically, the lesion is a cyst containing keratins. About one half of the cyst showed features of epidermal cyst consisting of mature squamous epithelium with granular layers. The other one half showed SK-like epidermal proliferation. The SK-like area showed basaloid cell proliferation with pseudohorn cysts. No significant atypia was noted. Many eosinophilic cytoplasmic inclusion bodies were noted in the SK-like area. Immunohistochemically, the SK-like area was positive for pancytokeratin AE1/3, pancytokeratin CAM5.2, p63, and Ki-67 (labeling=8%) and HPV, but negative for p53. The pathological diagnosis was SIC.

  11. Expression, purification and characterization of a full-length recombinant HIV-1 Vpu from inclusion bodies.

    Science.gov (United States)

    Njengele, Zikhona; Kleynhans, Ronel; Sayed, Yasien; Mosebi, Salerwe

    2016-12-01

    Vpu is one of four accessory proteins encoded by human immunodeficiency virus type I (HIV-1). Vpu modulates the expression of several cellular restriction factors within the HIV-1 infected cell including CD4, CD74, the bone marrow stromal antigen 2 (BST-2) and NK-T-and-B antigen. The interaction of HIV-1 Vpu with these proteins interferes with the innate immune response directed against HIV-1; thereby promoting viral persistence. The involvement of HIV-1 Vpu in manipulating the cellular environment in ways that favor viral replication makes it an attractive target for anti-HIV drug intervention. This paper describes the over-expression and purification of a soluble HIV-1 Vpu from inclusion bodies by ion-exchange chromatography, allowing production of 6 mg of highly purified protein (>95% purity) per 10 mg of pelleted cells obtained from 1 L of bacterial culture. Far-UV circular dichroism showed that the recombinant protein is folded and retained its secondary structure. Moreover, using ELISA, known HIV-1 Vpu binding partners, BST-2 and CD74, showed that the refolded purified protein is functional or at least assumes a conformation that is capable of binding these putative binding partners. To our knowledge, this is the first report of the purification and successful solubilization of full-length, wild-type HIV-1 Vpu from inclusion bodies in Escherichia coli. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Muscle sonography in six patients with hereditary inclusion body myopathy

    Energy Technology Data Exchange (ETDEWEB)

    Adler, Ronald S. [Weill Medical College of Cornell University, Division of Ultrasound and Body Imaging, Hospital for Special Surgery, New York, NY (United States); Garolfalo, Giovanna [Ospedale Residenza Sanitaria Riabilitativa, Fermo, AP (Italy); Paget, Stephen; Kagen, Lawrence [Weill Medical College of Cornell University, Rheumatology Division, Hospital for Special Surgery, New York, NY (United States)

    2008-01-15

    To evaluate the morphological changes of muscle with sonography in six patients affected by hereditary inclusion body myopathy (HIBM). We studied a group of six Persian Jews diagnosed with HIBM. All were homozygous for the GNE mutation M712T. Ultrasonographic examinations of the quadriceps femoris and hamstring muscle groups were performed. A follow-up ultrasound examination was performed, after an interval of 3 years, in four of these patients. Muscles were assessed subjectively as to echogenicity, determined by gray-scale assessment, and loss of normal muscle morphology. Power Doppler sonography (PDS) was used to assess vascularity. A sonographic finding of central atrophy and peripheral sparing resulting in a target-like appearance was noted in the hamstring compartment of all six patients. The quadriceps compartment also showed involvement of the rectus femoris of all patients, which, in some cases, was the only muscle involved in the quadriceps. Vascularity was markedly reduced in the affected areas, with blood flow demonstrated in the peripherally spared areas. The severity of atrophy increased with disease duration. In this case series, we describe a new sonographic finding as well as document progression of HIBM disease, which has generally been described as quadriceps sparing. The myopathic target lesion, as well as isolated rectus femoris atrophy, may provide a useful adjunct to disease diagnosis. (orig.)

  13. The proteomic profile of hereditary inclusion body myopathy.

    Directory of Open Access Journals (Sweden)

    Ilan Sela

    Full Text Available Hereditary inclusion body myopathy (HIBM is an adult onset, slowly progressive distal and proximal myopathy. Although the causing gene, GNE, encodes for a key enzyme in the biosynthesis of sialic acid, its primary function in HIBM remains unknown. The goal of this study was to unravel new clues on the biological pathways leading to HIBM by proteomic comparison. Muscle cultures and biopsies were analyzed by two dimensional gel electrophoresis (2-DE and the same biopsy extracts by isobaric tag for relative and absolute quantitation (iTRAQ. Proteins that were differentially expressed in all HIBM specimens versus all controls in each analysis were identified by mass spectrometry. The muscle cultures 2-DE analysis yielded 41 such proteins, while the biopsies 2-DE analysis showed 26 differentially expressed proteins. Out of the 400 proteins identified in biopsies by iTRAQ, 41 showed altered expression. In spite of the different nature of specimens (muscle primary cultures versus muscle biopsies and of the different methods applied (2D gels versus iTRAQ the differentially expressed proteins identified in each of the three analyses where related mainly to the same pathways, ubiquitination, stress response and mitochondrial processes, but the most robust cluster (30% was assigned to cytoskeleton and sarcomere organization. Taken together, these findings indicate a possible novel function of GNE in the muscle filamentous apparatus that could be involved in the pathogenesis of HIBM.

  14. Ultrastructure of inclusion bodies in annulus cells in the degenerating human intervertebral disc.

    Science.gov (United States)

    Gruber, H E; Hanley, E N

    2009-06-01

    The rough endoplasmic reticulum (rER) of the cell has an architectural editing function that checks whether protein structure and three-dimensional assembly have occurred properly prior to export of newly synthesized material out of the cell. If these have been faulty, the material is retained within the rER as an inclusion body. Inclusion bodies have been identified previously in chondrocytes and osteoblasts in chondrodysplasias and osteogenesis imperfecta. Inclusion bodies in intervertebral disc cells, however, have only recently been recognized. Our objectives were to use transmission electron microscopy to analyze more fully inclusion bodies in the annulus pulposus and to study the extracellular matrix (ECM) surrounding cells containing inclusion bodies. ECM frequently encapsulated cells with inclusion bodies, and commonly contained prominent banded aggregates of Type VI collagen. Inclusion body material had several morphologies, including relatively smooth, homogeneous material, or a rougher, less homogeneous feature. Such findings expand our knowledge of the fine structure of the human disc cell and ECM during disc degeneration, and indicate the potential utility of ultrastructural identification of discs with intracellular inclusion bodies as a screening method for molecular studies directed toward identification of defective gene products in degenerating discs.

  15. Luminescent conjugated oligothiophenes for sensitive fluorescent assignment of protein inclusion bodies.

    Science.gov (United States)

    Klingstedt, Therése; Blechschmidt, Cristiane; Nogalska, Anna; Prokop, Stefan; Häggqvist, Bo; Danielsson, Olof; Engel, W King; Askanas, Valerie; Heppner, Frank L; Nilsson, K Peter R

    2013-03-18

    Small hydrophobic ligands identifying intracellular protein deposits are of great interest, as protein inclusion bodies are the pathological hallmark of several degenerative diseases. Here we report that fluorescent amyloid ligands, termed luminescent conjugated oligothiophenes (LCOs), rapidly and with high sensitivity detect protein inclusion bodies in skeletal muscle tissue from patients with sporadic inclusion body myositis (s-IBM). LCOs having a conjugated backbone of at least five thiophene units emitted strong fluorescence upon binding, and showed co-localization with proteins reported to accumulate in s-IBM protein inclusion bodies. Compared with conventional amyloid ligands, LCOs identified a larger fraction of immunopositive inclusion bodies. When the conjugated thiophene backbone was extended with terminal carboxyl groups, the LCO revealed striking spectral differences between distinct protein inclusion bodies. We conclude that 1) LCOs are sensitive, rapid and powerful tools for identifying protein inclusion bodies and 2) LCOs identify a wider range of protein inclusion bodies than conventional amyloid ligands. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Preparative Protein Production from Inclusion Bodies and Crystallization: A Seven-Week Biochemistry Sequence

    Science.gov (United States)

    Peterson, Megan J.; Snyder, W. Kalani; Westerman, Shelley; McFarland, Benjamin J.

    2011-01-01

    We describe how to produce and purify proteins from "Escherichia coli" inclusion bodies by adapting versatile, preparative-scale techniques to the undergraduate laboratory schedule. This 7-week sequence of experiments fits into an annual cycle of research activity in biochemistry courses. Recombinant proteins are expressed as inclusion bodies,…

  17. Howell-Jolly body-like inclusions in the neutrophils of patients with acquired immunodeficiency syndrome.

    Science.gov (United States)

    Slagel, D D; Lager, D J; Dick, F R

    1994-04-01

    The authors have observed discrete, densely basophilic inclusions in the peripheral blood neutrophils of 10 patients with acquired immunodeficiency syndrome (AIDS) and two additional patients without documented evidence of human immunodeficiency virus seropositivity. These inclusions were seen in 3 of 25 (12%) patients with AIDS in a retrospective review. Gram, periodic acid-Schiff, and Gomori methenamine silver stains failed to demonstrate staining of these bodies; however, the Feulgen reaction was positive. The inclusions may represent "apoptotic" nuclear fragments and thus may be similar to Howell-Jolly bodies of normoblasts in abnormal erythropoiesis. An association of these inclusions with nucleoside analogue antiviral therapy is indicated. These inclusions must be differentiated from intracytoplasmic infectious agents, Döhle bodies, and inclusions of inherited disorders.

  18. Should body image programs be inclusive? A focus group study of college students.

    Science.gov (United States)

    Ciao, Anna C; Ohls, Olivia C; Pringle, Kevin D

    2018-01-01

    Most evidence-based body image programs for college students (e.g., the Body Project) are designed for female-only audiences, although body dissatisfaction is not limited to female-identified individuals. Furthermore, programs do not explicitly discuss diversity, although individuals with marginalized gender, racial, and sexual identities may be particularly vulnerable to body image disturbances. Making programs more inclusive may increase their disseminability. This qualitative study examined the feasibility of adapting the Body Project for universal and inclusive use with college students. Participants (N = 36; M age = 21.66 years; 73% female-identified; 20% sexual minority; 23% racial minority) attended one of five semi-structured focus groups to explore the inclusivity of appearance-based cultural norms using adapted Body Project activities and discuss the feasibility of universal and inclusive interventions. Inductive qualitative content analysis with three-rater consensus identified focus group themes. There was consensus that inclusive interventions could have a positive impact (broadening perspectives, normalizing body image concerns, increasing awareness) despite potential barriers (poor diversity representation, vulnerability). There was strong consensus regarding advice for facilitating inclusive interventions (e.g., skilled facilitation, education, increasing diversity). Results suggest that inclusive body image programs are desirable and provide a framework for creating the EVERYbody Project, a program for more universal audiences. © 2017 Wiley Periodicals, Inc.

  19. TDP-43 inclusion bodies formed in bacteria are structurally amorphous, non-amyloid and inherently toxic to neuroblastoma cells.

    Directory of Open Access Journals (Sweden)

    Claudia Capitini

    Full Text Available Accumulation of ubiquitin-positive, tau- and α-synuclein-negative intracellular inclusions of TDP-43 in the central nervous system represents the major hallmark correlated to amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitin-positive inclusions. Such inclusions have variably been described as amorphous aggregates or more structured deposits having an amyloid structure. Following the observations that bacterial inclusion bodies generally consist of amyloid aggregates, we have overexpressed full-length TDP-43 and C-terminal TDP-43 in E. coli, purified the resulting full-length and C-terminal TDP-43 containing inclusion bodies (FL and Ct TDP-43 IBs and subjected them to biophysical analyses to assess their structure/morphology. We show that both FL and Ct TDP-43 aggregates contained in the bacterial IBs do not bind amyloid dyes such as thioflavin T and Congo red, possess a disordered secondary structure, as inferred using circular dichroism and infrared spectroscopies, and are susceptible to proteinase K digestion, thus possessing none of the hallmarks for amyloid. Moreover, atomic force microscopy revealed an irregular structure for both types of TDP-43 IBs and confirmed the absence of amyloid-like species after proteinase K treatment. Cell biology experiments showed that FL TDP-43 IBs were able to impair the viability of cultured neuroblastoma cells when added to their extracellular medium and, more markedly, when transfected into their cytosol, where they are at least in part ubiquitinated and phosphorylated. These data reveal an inherently high propensity of TDP-43 to form amorphous aggregates, which possess, however, an inherently high ability to cause cell dysfunction. This indicates that a gain of toxic function caused by TDP-43 deposits is effective in TDP-43 pathologies, in addition to possible loss of function mechanisms originating from the cellular mistrafficking of the protein.

  20. Formation of distinct inclusion bodies by inhibition of ubiquitin-proteasome and autophagy-lysosome pathways.

    Science.gov (United States)

    Lee, Junho; Yang, Kyu-Hwan; Joe, Cheol O; Kang, Seok-Seong

    2011-01-14

    Accumulation of misfolded proteins is caused by the impairment of protein quality control systems, such as ubiquitin-proteasome pathway (UPP) and autophagy-lysosome pathway (ALP). In this study, the formation of inclusion bodies was examined after the blockade of UPP and/or ALP in A549 cells. UPP inhibition induced a single and large inclusion body localized in microtubule-organizing center. Interestingly, however, ALP inhibition generated dispersed small inclusion bodies in the cytoplasm. Tuberous sclerosis complex 2 was selectively accumulated in the inclusion bodies of UPP-inhibited cells, but not those of ALP-inhibited cells. Blockade of transcription and translation entirely inhibited the formation of inclusion body induced by UPP inhibition, but partially by ALP inhibition. Moreover, the simultaneous inhibition of two protein catabolic pathways independently developed two distinct inclusion bodies within a single cell. These findings clearly demonstrated that dysfunction of each catabolic pathway induced formation and accumulation of unique inclusion bodies on the basis of morphology, localization and formation process in A549 cells. Copyright © 2010 Elsevier Inc. All rights reserved.

  1. Refolding techniques for recovering biologically active recombinant proteins from inclusion bodies.

    Science.gov (United States)

    Yamaguchi, Hiroshi; Miyazaki, Masaya

    2014-02-20

    Biologically active proteins are useful for studying the biological functions of genes and for the development of therapeutic drugs and biomaterials in a biotechnology industry. Overexpression of recombinant proteins in bacteria, such as Escherichia coli, often results in the formation of inclusion bodies, which are protein aggregates with non-native conformations. As inclusion bodies contain relatively pure and intact proteins, protein refolding is an important process to obtain active recombinant proteins from inclusion bodies. However, conventional refolding methods, such as dialysis and dilution, are time consuming and, often, recovered yields of active proteins are low, and a trial-and-error process is required to achieve success. Recently, several approaches have been reported to refold these aggregated proteins into an active form. The strategies largely aim at reducing protein aggregation during the refolding procedure. This review focuses on protein refolding techniques using chemical additives and laminar flow in microfluidic chips for the efficient recovery of active proteins from inclusion bodies.

  2. Application of an E. coli signal sequence as a versatile inclusion body tag

    NARCIS (Netherlands)

    Jong, Wouter S.P.; Vikström, David; Houben, Diane; Berg van Saparoea, H. Bart; de Gier, Jan Willem; Luirink, Joen

    2017-01-01

    Background: Heterologous protein production in Escherichia coli often suffers from bottlenecks such as proteolytic degradation, complex purification procedures and toxicity towards the expression host. Production of proteins in an insoluble form in inclusion bodies (IBs) can alleviate these

  3. Inclusion-body myositis, a multifactorial muscle disease associated with aging: current concepts of pathogenesis.

    Science.gov (United States)

    Askanas, Valerie; Engel, W King

    2007-11-01

    Sporadic inclusion-body myositis, the most common muscle disease of older persons, has no known cause or persistently beneficial treatment. The unfolding pathogenesis could lead to new treatment strategies and it is now of growing interest among clinicians and basic scientists. About 100 papers related to the subject were published in 2006 and the first part of 2007 (we cite only articles most relevant to this review). This review focuses on the current concepts of the pathogenesis of sporadic inclusion-body myositis. Both degeneration and mononuclear-cell inflammation are components of the pathology, but how each relates to the pathogenesis remains unclear. We suggest that an intramuscle fiber degenerative component is primary, leading to muscle-fiber destruction, while the lymphocytic inflammatory component may only slightly contribute to sporadic inclusion-body myositis muscle-fiber damage. Intracellular accumulation of amyloid-beta precursor protein, amyloid-beta, and amyloid-beta oligomers in an aging muscle-fiber cellular milieu, and other abnormalities, appear to be key pathogenic factors. We summarize intracellular molecular events and their consequences, and correlate findings in sporadic inclusion-body myositis muscle biopsies with inclusion-body myositis experimental models in tissue culture and in transgenic mice. Treatment of sporadic inclusion-body myositis remains a challenge. Antiinflammatory approaches used so far are without major or enduring benefit. Possible new treatment avenues are suggested.

  4. Adenovirus-like inclusion body hepatitis in a flock of broiler chickens in Kermanshah province, Iran.

    Science.gov (United States)

    Rahimi, Morad; Minoosh Siavosh Haghighi, Zahra

    2015-01-01

    Inclusion body hepatitis (IBH) has been reported in many countries in the world. The IBH or similar cases characterized by hepatitis and presence of intra-nuclear inclusion bodies in hepatocytes have not been reported in broiler chickens in Iran. This is the first report on outbreak of adenovirus-like inclusion body hepatitis in Iran. On October 2012, an onset of high acute mortality in a flock of 2 day-old broiler chickens was reported to the Veterinary Clinic, Faculty of Veterinary Medicine, Razi University, Kermanshah, Iran. The birds showed lethargy, huddling, ruffled feathers, and inappetence. At necropsy the livers were the primary organ affected which were enlarged, pale yellow with necrotic foci and multiple petechial hemorrhages. Tissue samples of liver, kidneys and heart were fixed in 10% buffered formalin. They were processed and stained with hematoxylin and eosin for histopathological studies. Significant microscopic lesions were seen in the livers. Large eosinophilic intra-nuclear inclusion bodies were seen in hepatocytes. Based on the acute high mortality, age of the broilers, gross lesions and histopathological findings (especially intra-nuclear inclusion bodies), the condition was diagnosed as adenovirus-like inclusion body hepatitis.

  5. Alternative preparation of inclusion bodies excludes interfering non-protein contaminants and improves the yield of recombinant proinsulin.

    Science.gov (United States)

    Mackin, Robert B

    2014-01-01

    The goal of simple, high-yield expression and purification of recombinant human proinsulin has proven to be a considerable challenge. First, proinsulin forms inclusion bodies during bacterial expression. While this phenomenon can be exploited as a capture step, conventionally prepared inclusion bodies contain significant amounts of non-protein contaminants that interfere with subsequent chromatographic purification. Second, the proinsulin molecules within the inclusion bodies are incorrectly folded, and likely cross-linked to one another, making it difficult to quantify the amount of expressed proinsulin. Third, proinsulin is an intermediate between the initial product of ribosomal translation (preproinsulin) and the final product secreted by pancreatic beta cells (insulin). Therefore, to be efficiently produced in bacteria, it must be produced as an N-terminally extended fusion protein, which has to be converted to authentic proinsulin during the purification scheme. To address all three of these problems, while simultaneously streamlining the procedure and increasing the yield of recombinant proinsulin, we have made three substantive modifications to our previous method for producing proinsulin:.•Conditions for the preparation of inclusion bodies have been altered so contaminants that interfere with semi-preparative reversed-phase chromatography are excluded while the proinsulin fusion protein is retained at high yield.•Aliquots are taken following important steps in the procedure and the quantity of proinsulin-related polypeptide in the sample is compared to the amount present prior to that step.•Final purification is performed using a silica-based reversed-phase matrix in place of a polystyrene-divinylbenzene-based matrix.

  6. Analysis of bacterial-surface-specific antibodies in body fluids using bacterial flow cytometry.

    Science.gov (United States)

    Moor, Kathrin; Fadlallah, Jehane; Toska, Albulena; Sterlin, Delphine; Balmer, Maria L; Macpherson, Andrew J; Gorochov, Guy; Larsen, Martin; Slack, Emma

    2016-08-01

    Antibacterial antibody responses that target surfaces of live bacteria or secreted toxins are likely to be relevant in controlling bacterial pathogenesis. The ability to specifically quantify bacterial-surface-binding antibodies is therefore highly attractive as a quantitative correlate of immune protection. Here, binding of antibodies from various body fluids to pure-cultured live bacteria is made visible with fluorophore-conjugated secondary antibodies and measured by flow cytometry. We indicate the necessary controls for excluding nonspecific binding and also demonstrate a cross-adsorption technique for determining the extent of cross-reactivity. This technique has numerous advantages over standard ELISA and western blotting techniques because of its independence from scaffold binding, exclusion of cross-reactive elements from lysed bacteria and ability to visualize bacterial subpopulations. In addition, less than 10(5) bacteria and less than 10 μg of antibody are required per sample. The technique requires 3-4 h of hands-on experimentation and analysis. Moreover, it can be combined with automation and mutliplexing for high-throughput applications.

  7. A bibliography of high energy two-body and inclusive scattering data

    International Nuclear Information System (INIS)

    Gault, F.D.; Read, B.J.; Roberts, R.G.

    1977-09-01

    A bibliography is presented of the data on high energy two-body and quasi-two-body final state scattering processes. This updated edition also covers one and two-particle inclusive production. It contains references to those published papers whose main purpose is to provide data on high energy two-body and inclusive hadronic scattering cross-sections rather than just properties of the produced particles. It covers the leading high energy physics journals and the period up to June 1977. The entries are grouped by process in the order indicated in the Table of Contents, and an author index is also provided. (author)

  8. Application of preparative disk gel electrophoresis for antigen purification from inclusion bodies.

    Science.gov (United States)

    Okegawa, Yuki; Koshino, Masanori; Okushima, Teruya; Motohashi, Ken

    2016-02-01

    Specific antibodies are a reliable tool to examine protein expression patterns and to determine the protein localizations within cells. Generally, recombinant proteins are used as antigens for specific antibody production. However, recombinant proteins from mammals and plants are often overexpressed as insoluble inclusion bodies in Escherichia coli. Solubilization of these inclusion bodies is desirable because soluble antigens are more suitable for injection into animals to be immunized. Furthermore, highly purified proteins are also required for specific antibody production. Plastidic acetyl-CoA carboxylase (ACCase: EC 6.4.1.2) from Arabidopsis thaliana, which catalyzes the formation of malonyl-CoA from acetyl-CoA in chloroplasts, formed inclusion bodies when the recombinant protein was overexpressed in E. coli. To obtain the purified protein to use as an antigen, we applied preparative disk gel electrophoresis for protein purification from inclusion bodies. This method is suitable for antigen preparation from inclusion bodies because the purified protein is recovered as a soluble fraction in electrode running buffer containing 0.1% sodium dodecyl sulfate that can be directly injected into immune animals, and it can be used for large-scale antigen preparation (several tens of milligrams). Copyright © 2015 Elsevier Inc. All rights reserved.

  9. FLOW RESTRICTED RESISTANCE TRAINING ATTENUATES MYOSTATIN GENE EXPRESSION IN A PATIENT WITH INCLUSION BODY MYOSITIS

    Directory of Open Access Journals (Sweden)

    A.R. Santos

    2014-07-01

    Full Text Available Inclusion body myositis is a rare idiopathic inflammatory myopathy that produces extreme muscle weakness. Blood flow restricted resistance training has been shown to improve muscle strength and muscle hypertrophy in inclusion body myositis. Objective: The aim of this study was to evaluate the effects of a resistance training programme on the expression of genes related to myostatin (MSTN signalling in one inclusion body myositis patient. Methods: A 65-year-old man with inclusion body myositis underwent blood flow restricted resistance training for 12 weeks. The gene expression of MSTN, follistatin, follistatin-like 3, activin II B receptor, SMAD-7, MyoD, FOXO-3, and MURF-2 was quantified. Results: After 12 weeks of training, a decrease (25% in MSTN mRNA level was observed, whereas follistatin and follistatin-like 3 gene expression increased by 40% and 70%, respectively. SMAD-7 mRNA level was augmented (20%. FOXO-3 and MURF-2 gene expression increased by 40% and 20%, respectively. No change was observed in activin II B receptor or MyoD gene expression. Conclusions: Blood flow restricted resistance training attenuated MSTN gene expression and also increased expression of myostatin endogenous inhibitors. Blood flow restricted resistance training evoked changes in the expression of genes related to MSTN signalling pathway that could in part explain the muscle hypertrophy previously observed in a patient with inclusion body myositis.

  10. Microfluidic chips with multi-junctions: an advanced tool in recovering proteins from inclusion bodies.

    Science.gov (United States)

    Yamaguchi, Hiroshi; Miyazaki, Masaya

    2015-01-01

    Active recombinant proteins are used for studying the biological functions of genes and for the development of therapeutic drugs. Overexpression of recombinant proteins in bacteria often results in the formation of inclusion bodies, which are protein aggregates with non-native conformations. Protein refolding is an important process for obtaining active recombinant proteins from inclusion bodies. However, the conventional refolding method of dialysis or dilution is time-consuming and recovered active protein yields are often low, and a cumbersome trial-and-error process is required to achieve success. To circumvent these difficulties, we used controllable diffusion through laminar flow in microchannels to regulate the denaturant concentration. This method largely aims at reducing protein aggregation during the refolding procedure. This Commentary introduces the principles of the protein refolding method using microfluidic chips and the advantage of our results as a tool for rapid and efficient recovery of active recombinant proteins from inclusion bodies.

  11. Kinetics of Inclusion Body Formation and Its Correlation with the Characteristics of Protein Aggregates in Escherichia coli

    Science.gov (United States)

    Upadhyay, Arun K.; Murmu, Aruna; Singh, Anupam; Panda, Amulya K.

    2012-01-01

    The objective of the research was to understand the structural determinants governing protein aggregation into inclusion bodies during expression of recombinant proteins in Escherichia coli. Recombinant human growth hormone (hGH) and asparaginase were expressed as inclusion bodies in E.coli and the kinetics of aggregate formation was analyzed in details. Asparaginase inclusion bodies were of smaller size (200 nm) and the size of the aggregates did not increase with induction time. In contrast, the seeding and growth behavior of hGH inclusion bodies were found to be sequential, kinetically stable and the aggregate size increased from 200 to 800 nm with induction time. Human growth hormone inclusion bodies showed higher resistance to denaturants and proteinase K degradation in comparison to those of asparaginase inclusion bodies. Asparaginase inclusion bodies were completely solubilized at 2–3 M urea concentration and could be refolded into active protein, whereas 7 M urea was required for complete solubilization of hGH inclusion bodies. Both hGH and asparaginase inclusion bodies showed binding with amyloid specific dyes. In spite of its low β-sheet content, binding with dyes was more prominent in case of hGH inclusion bodies than that of asparaginase. Arrangements of protein molecules present in the surface as well as in the core of inclusion bodies were similar. Hydrophobic interactions between partially folded amphiphillic and hydrophobic alpha-helices were found to be one of the main determinants of hGH inclusion body formation. Aggregation behavior of the protein molecules decides the nature and properties of inclusion bodies. PMID:22479486

  12. Kinetics of inclusion body formation and its correlation with the characteristics of protein aggregates in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Arun K Upadhyay

    Full Text Available The objective of the research was to understand the structural determinants governing protein aggregation into inclusion bodies during expression of recombinant proteins in Escherichia coli. Recombinant human growth hormone (hGH and asparaginase were expressed as inclusion bodies in E.coli and the kinetics of aggregate formation was analyzed in details. Asparaginase inclusion bodies were of smaller size (200 nm and the size of the aggregates did not increase with induction time. In contrast, the seeding and growth behavior of hGH inclusion bodies were found to be sequential, kinetically stable and the aggregate size increased from 200 to 800 nm with induction time. Human growth hormone inclusion bodies showed higher resistance to denaturants and proteinase K degradation in comparison to those of asparaginase inclusion bodies. Asparaginase inclusion bodies were completely solubilized at 2-3 M urea concentration and could be refolded into active protein, whereas 7 M urea was required for complete solubilization of hGH inclusion bodies. Both hGH and asparaginase inclusion bodies showed binding with amyloid specific dyes. In spite of its low β-sheet content, binding with dyes was more prominent in case of hGH inclusion bodies than that of asparaginase. Arrangements of protein molecules present in the surface as well as in the core of inclusion bodies were similar. Hydrophobic interactions between partially folded amphiphillic and hydrophobic alpha-helices were found to be one of the main determinants of hGH inclusion body formation. Aggregation behavior of the protein molecules decides the nature and properties of inclusion bodies.

  13. Intracellular organisation of polyhydroxyalkanoate inclusion bodies: a role for small angle neutron scattering?

    International Nuclear Information System (INIS)

    Foster, L.J.R.; Holden, P.J.; Garvey, C.J.; Russell, R.A.; Stone, D.J.M.

    2003-01-01

    Full text: Polyhydroxyalkanoates (PHAs) are a diverse family of bacterially produced biopolyesters. Their biodegradability, and in some cases biocompatibility, suggest applications ranging from bioplastics to biomedical implantation devices. Despite extensive interest in their production and potential applications, little is known about their intracellular organisation. Microbial PHAs are synthesised by microorganisms under conditions of nutrient stress and can comprise up to 90% of the dry cell mass. The formation and organisation of these PHA inclusion bodies requires clarification. Such investigations have important implications for the biotechnological production of PHAs in microbes and other organisms, for downstream processing and in vitro precision polymerisation. Morphological and biochemical evidence supports two different models for the intracellular organisation of PHAs. Steinbuchel and coworkers propose a simple model of amorphous PHA enclosed by a single protein membrane consisting of structural proteins (PHAsins) and enzymes responsible for synthesis and degradation. In contrast, Fuller and coworkers have theorised a more complex system of PHA encompassed by a PHAsin bilayer separated by phospholipid. The polymerase and depolymerase enzymes are proposed to be associated with an incomplete inner PHAsin layer. It may be that such models are genera or species specific, since both proposals were derived from research on different species producing different types of PHA. Our initial investigations have focussed on in vivo deuteration of polyhydroxyoctanoate, produced by Pseudomonas oleovorans, both in fermentation on natural and deuterated substrates and during Small Angle Neutron Scattering by whole cells using AUSANS. The nature of the structural questions and our preliminary findings including contrast variation data will be discussed

  14. Direct Conversion of an Enzyme from Native-like to Amyloid-like Aggregates within Inclusion Bodies.

    Science.gov (United States)

    Elia, Francesco; Cantini, Francesca; Chiti, Fabrizio; Dobson, Christopher Martin; Bemporad, Francesco

    2017-06-20

    The acylphosphatase from Sulfolobus solfataricus (Sso AcP) is a globular protein able to aggregate in vitro from a native-like conformational ensemble without the need for a transition across the major unfolding energy barrier. This process leads to the formation of assemblies in which the protein retains its native-like structure, which subsequently convert into amyloid-like aggregates. Here, we investigate the mechanism by which Sso AcP aggregates in vivo to form bacterial inclusion bodies after expression in E. coli. Shortly after the initiation of expression, Sso AcP is incorporated into inclusion bodies as a native-like protein, still exhibiting small but significant enzymatic activity. Additional experiments revealed that this overall process of aggregation is enhanced by the presence of the unfolded N-terminal region of the sequence and by destabilization of the globular segment of the protein. At later times, the Sso AcP molecules in the inclusion bodies lose their native-like properties and convert into β-sheet-rich amyloid-like structures, as indicated by their ability to bind thioflavin T and Congo red. These results show that the aggregation behavior of this protein is similar in vivo to that observed in vitro, and that, at least for a predominant part of the protein population, the transition from a native to an amyloid-like structure occurs within the aggregate state. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  15. Human Metapneumovirus Induces Formation of Inclusion Bodies for Efficient Genome Replication and Transcription.

    Science.gov (United States)

    Cifuentes-Muñoz, Nicolás; Branttie, Jean; Slaughter, Kerri Beth; Dutch, Rebecca Ellis

    2017-12-15

    Human metapneumovirus (HMPV) causes significant upper and lower respiratory disease in all age groups worldwide. The virus possesses a negative-sense single-stranded RNA genome of approximately 13.3 kb encapsidated by multiple copies of the nucleoprotein (N), giving rise to helical nucleocapsids. In addition, copies of the phosphoprotein (P) and the large RNA polymerase (L) decorate the viral nucleocapsids. After viral attachment, endocytosis, and fusion mediated by the viral glycoproteins, HMPV nucleocapsids are released into the cell cytoplasm. To visualize the subsequent steps of genome transcription and replication, a fluorescence in situ hybridization (FISH) protocol was established to detect different viral RNA subpopulations in infected cells. The FISH probes were specific for detection of HMPV positive-sense RNA (+RNA) and viral genomic RNA (vRNA). Time course analysis of human bronchial epithelial BEAS-2B cells infected with HMPV revealed the formation of inclusion bodies (IBs) from early times postinfection. HMPV IBs were shown to be cytoplasmic sites of active transcription and replication, with the translation of viral proteins being closely associated. Inclusion body formation was consistent with an actin-dependent coalescence of multiple early replicative sites. Time course quantitative reverse transcription-PCR analysis suggested that the coalescence of inclusion bodies is a strategy to efficiently replicate and transcribe the viral genome. These results provide a better understanding of the steps following HMPV entry and have important clinical implications. IMPORTANCE Human metapneumovirus (HMPV) is a recently discovered pathogen that affects human populations of all ages worldwide. Reinfections are common throughout life, but no vaccines or antiviral treatments are currently available. In this work, a spatiotemporal analysis of HMPV replication and transcription in bronchial epithelial cell-derived immortal cells was performed. HMPV was shown to

  16. Comparative study to develop a single method for retrieving wide class of recombinant proteins from classical inclusion bodies.

    Science.gov (United States)

    Padhiar, Arshad Ahmed; Chanda, Warren; Joseph, Thomson Patrick; Guo, Xuefang; Liu, Min; Sha, Li; Batool, Samana; Gao, Yifan; Zhang, Wei; Huang, Min; Zhong, Mintao

    2018-03-01

    The formation of inclusion bodies (IBs) is considered as an Achilles heel of heterologous protein expression in bacterial hosts. Wide array of techniques has been developed to recover biochemically challenging proteins from IBs. However, acquiring the active state even from the same protein family was found to be an independent of single established method. Here, we present a new strategy for the recovery of wide sub-classes of recombinant protein from harsh IBs. We found that numerous methods and their combinations for reducing IB formation and producing soluble proteins were not effective, if the inclusion bodies were harsh in nature. On the other hand, different practices with mild solubilization buffers were able to solubilize IBs completely, yet the recovery of active protein requires large screening of refolding buffers. With the integration of previously reported mild solubilization techniques, we proposed an improved method, which comprised low sarkosyl concentration, ranging from 0.05 to 0.1% coupled with slow freezing (- 1 °C/min) and fast thaw (room temperature), resulting in greater solubility and the integrity of solubilized protein. Dilution method was employed with single buffer to restore activity for every sub-class of recombinant protein. Results showed that the recovered protein's activity was significantly higher compared with traditional solubilization/refolding approach. Solubilization of IBs by the described method was proved milder in nature, which restored native-like conformation of proteins within IBs.

  17. Myositis with endomysial cell invasion indicates inclusion body myositis even if other criteria are not fulfilled

    NARCIS (Netherlands)

    van de Vlekkert, J; Hoogendijk, J E; de Visser, M

    The objective of this study was to investigate if patients with endomysial mononuclear cell infiltrates invading non-necrotic fibers have a disease course consistent with inclusion body myositis (IBM), irrespective of other histopathological and clinical characteristics. All patients with a muscle

  18. Autoantibodies to cytosolic 5'-nucleotidase 1A in inclusion body myositis

    NARCIS (Netherlands)

    Pluk, H.; Hoeve, B.J.A. van; Dooren, S.H. van; Stammen-Vogelzangs, J.; Heijden, A. van der; Schelhaas, H.J.; Verbeek, M.M.; Badrising, U.A.; Arnardottir, S.; Gheorghe, K.; Lundberg, I.E.; Boelens, W.C.; Engelen, B.G.M. van; Pruijn, Ger

    2013-01-01

    OBJECTIVE: Sporadic inclusion body myositis (sIBM) is an inflammatory myopathy characterized by both degenerative and autoimmune features. In contrast to other inflammatory myopathies, myositis-specific autoantibodies had not been found in sIBM patients until recently. We used human skeletal muscle

  19. Dean vortex membrane microfiltration and diafiltration of rBDNF E. coli inclusion bodies

    NARCIS (Netherlands)

    Schutyser, M.A.I.; Rupp, R.; Wideman, J.; Belfort, G.

    2002-01-01

    Cross-flow microfiltration (CMF) and diafiltration were used to concentrate and purify recombinant Brain-Derived Neutrophic Factor (rBDNF) inclusion bodies from an E. coli cell suspension and a homogenized E. coli cell suspension (homogenate/lysate). Although these processes have been tested

  20. Detection of novel divergent arenaviruses in boid snakes with inclusion body disease in The Netherlands

    NARCIS (Netherlands)

    R. Bodewes (Rogier); M.J.L. Kik (Marja); V.S. Stalin Raj; C.M.E. Schapendonk (Claudia); B.L. Haagmans (Bart); S.L. Smits (Saskia); A.D.M.E. Osterhaus (Albert)

    2013-01-01

    textabstractArenaviruses are bi-segmented negative-stranded RNA viruses, which were until recently only detected in rodents and humans. Now highly divergent arenaviruses have been identified in boid snakes with inclusion body disease (IBD). Here, we describe the identification of a new species and

  1. A large planetary body inferred from diamond inclusions in a ureilite meteorite.

    Science.gov (United States)

    Nabiei, Farhang; Badro, James; Dennenwaldt, Teresa; Oveisi, Emad; Cantoni, Marco; Hébert, Cécile; El Goresy, Ahmed; Barrat, Jean-Alix; Gillet, Philippe

    2018-04-17

    Planetary formation models show that terrestrial planets are formed by the accretion of tens of Moon- to Mars-sized planetary embryos through energetic giant impacts. However, relics of these large proto-planets are yet to be found. Ureilites are one of the main families of achondritic meteorites and their parent body is believed to have been catastrophically disrupted by an impact during the first 10 million years of the solar system. Here we studied a section of the Almahata Sitta ureilite using transmission electron microscopy, where large diamonds were formed at high pressure inside the parent body. We discovered chromite, phosphate, and (Fe,Ni)-sulfide inclusions embedded in diamond. The composition and morphology of the inclusions can only be explained if the formation pressure was higher than 20 GPa. Such pressures suggest that the ureilite parent body was a Mercury- to Mars-sized planetary embryo.

  2. Refolding in high hydrostatic pressure of recombinant proteins from inclusion bodies in Escherichia Coli

    International Nuclear Information System (INIS)

    Balduino, Keli Nunes

    2009-01-01

    The expression of proteins as inclusion bodies in bacteria is a widely used alternative for production of recombinant protein. However, the aggregation is a problem often encountered during refolding of these proteins. High hydrostatic pressure are able to solubilise the inclusion bodies in the presence of low concentrations of denaturant reagents, encouraging refolding protein with high efficiency and reduce costs. This work aims to refolding of recombinant proteins expressed in Escherichia coli from inclusion bodies using high hydrostatic pressure. Three toxins, all featuring five or more disulfide bonds were studied: NXH8, Natterin 2 and Bothropstoxin 1. Suspensions of inclusion bodies of the three proteins were pressurized to 2000 bars for 16 hours. The buffers were optimized for refolding of the three proteins. The buffer used in the refolding of NXH8 was 50 mM Tris HCl, pH 9.0 with proportion of 1GSH: 4GSSG at a concentration of 6 mM and 2 M GdnHCl. Inclusion bodies were used in O.D. (A600nm) of 0.5. After refolding process, dialysis was performed at pH 7.0. The final yield of obtaining soluble NXH8 was 40% (28,6 mg of soluble NXH8/L of culture medium). The refolding of Bothropstoxin 1 was obtained in refolding buffer of Tris HCl 50 mM, pH 7,5 with proportion of 2 GSH: GSSG 3 and concentration of 3 mM and 1 M GdnHCl. Use with a suspension of O.D. (A600nm) of 0.5. The final yield of recovery of Bothropstoxin 1 refolded was 32% (9,2 mg of refolded Bothropstoxin 1/L of culture medium). The refolding of Natterin 2 was performed in the refolding buffer: 20 mM Tris HCl pH 9.0 at a ratio of 2 GSH: 3GSSG and concentration of 10 mM and 1 M GdnHCl and inclusion bodies O.D. (A600nm) of 6.0. The yield of Natterin 2 refolded was 20% (3,7 mg/L of culture medium). Physico-chemical and biological analysis were performed by SDS-PAGE, western blot, scanning electron microscopy, biological tests in vivo and in vitro and structural. The analysis conducted in NXH8 did not show

  3. 40 CFR 180.1149 - Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Inclusion bodies of the multi-nuclear....1149 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS... requirement of a tolerance. The microbial pest control agent inclusion bodies of the multi-nuclear...

  4. Intracytoplasmic inclusion bodies associated with vesicular ulcerative and necrotizing lesions of the digestive mucosa in fallow deer (Dama dama L.

    Directory of Open Access Journals (Sweden)

    Regina Diaz

    1990-09-01

    Full Text Available Intracytoplasmic epithelial inclusion bodies in the digestive mucosa of fallow deer (Dama dama L. were found to most probably be the result of an unspecific degenerative or post mortal change. There are reasons to believe that this is true also for the inclusion bodies found in reindeer, roe deer and moose.

  5. Isolation of a herpesvirus from an American kestrel with inclusion body disease.

    Science.gov (United States)

    Potgieter, L N; Kocan, A A; Kocan, K M

    1979-01-01

    A herpesvirus was isolated from the liver of a captive-bred American kestrel (Falco sparverius) which had died of inclusion body disease. Initial isolation was achieved in chicken embryo fibroblasts after three blind passages. Cell-adapted virus produced a distinct rounding of CEF cells within 24 to 48 h. Biologic and serologic tests suggested that the kestrel virus is similar to falcon herpesvirus and pigeon herpesvirus and is at least partially related to owl herpesvirus. However, serologic tests indicated that the kestrel herpesvirus is neither related to infectious laryngotracheitis virus nor to a herpesvirus from a psittacine bird; (Eupsittula canicularis) with Pacheco's parrot disease. This is the first report on the recovery of a herpesvirus similar to falcon herpesvirus from an American kestrel with naturally-occurring inclusion body disease, and on the serologic comparison between falcon herpesvirus and a psittacine herpesvirus.

  6. Association of inclusion body myositis with T cell large granular lymphocytic leukaemia

    DEFF Research Database (Denmark)

    Greenberg, Steven A; Pinkus, Jack L; Amato, Anthony A

    2016-01-01

    SEE HOHLFELD AND SCHULZE-KOOPS DOI101093/BRAIN/AWW053 FOR A SCIENTIFIC COMMENTARY ON THIS ARTICLE: Inclusion body myositis and T cell large granular lymphocytic leukaemia are rare diseases involving pathogenic cytotoxic CD8+ T cells. After encountering four patients with both disorders, we...... on follow-up testing in 15 patients a median of 350 days later. T cell aberrant loss of CD5 or gain of expression of CD16 and CD94 were common (19/42, 45%). In comparison, 2/15 (14%) age-matched patients with dermatomyositis, polymyositis, or necrotizing myopathy, and 0/20 (0%) age-matched healthy subjects...... prospectively screened 38 patients with inclusion body myositis for the presence of expanded large granular lymphocyte populations by standard clinical laboratory methods (flow cytometry, examination of blood smears, and T cell receptor gene rearrangements), and performed muscle immunohistochemistry for CD8, CD...

  7. Purification and refolding of a novel β-agarase from inclusion body of E. coli

    Science.gov (United States)

    Zhang, Li; Lu, Xinzhi; Han, Feng; Ma, Cuiping; Yu, Wengong

    2007-01-01

    β-agarase AgaB appears to represent a new family of glycoside hydrolase; it is structurally and functionally different from other known agarases. In the present study, AgaB was expressed with a temperature-inducible expression system in E. coli BL21 (DE3) as a fusion protein bearing a C-terminal hexahistidine tag. The protein existed mainly in the form of inclusion body. After being washed and solubilized, AgaB in inclusion body was denatured and purified to electrophoretic purity by immobilized metal affinity chromatography. The purified AgaB was then refolded using a simple pulse dilution method, and the refolded AgaB showed a high specific hydrolysis activity of about 1600 units /mg protein. Forty milligrams of refolded pure protein were obtained from 1L of culture.

  8. A simple strategy for the purification of native recombinant full-length human RPL10 protein from inclusion bodies.

    Science.gov (United States)

    Pereira, Larissa M; Silva, Luana R; Alves, Joseane F; Marin, Nélida; Silva, Flavio Sousa; Morganti, Ligia; Silva, Ismael D C G; Affonso, Regina

    2014-09-01

    The L10 ribosomal protein (RPL10) plays a role in the binding of the 60 S and 40 S ribosomal subunits and in mRNA translation. The evidence indicates that RPL10 also has multiple extra-ribosomal functions, including tumor suppression. Recently, the presence of RPL10 in prostate and ovarian cancers was evaluated, and it was demonstrated to be associated with autistic disorders and premature ovarian failure. In the present work, we successfully cloned and expressed full-length human RPL10 (hRPL10) protein and isolated inclusion bodies containing this protein that had formed under mild growth conditions. The culture produced 376mg of hRPL10 protein per liter of induced bacterial culture, of which 102.4mg was present in the soluble fraction, and 25.6mg was recovered at approximately 94% purity. These results were obtained using a two-step process of non-denaturing protein extraction from pelleted inclusion bodies. We studied the characteristics of this protein using circular dichroism spectroscopy and by monitoring the changes induced by the presence or absence of zinc ions using fluorescence spectrometry. The results demonstrated that the protein obtained using these non-conventional methods retained its secondary and tertiary structure. The conformational changes induced by the incorporation of zinc suggested that this protein could interact with Jun or the SH3 domain of c-yes. The results suggested that the strategy used to obtain hRPL10 is simple and could be applied to obtaining other proteins that are susceptible to degradation. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Columbid herpesvirus-1 in two Cooper's hawks (Accipiter cooperii) with fatal inclusion body disease.

    Science.gov (United States)

    Pinkerton, Marie E; Wellehan, James F X; Johnson, April J; Childress, April L; Fitzgerald, Scott D; Kinsel, Michael J

    2008-07-01

    We report two separate naturally occurring cases of fatal herpesviral disease in Cooper's Hawks (Accipiter cooperii). Gross lesions included splenomegaly and hepatomegaly, with diffuse pale mottling or scattered small white foci. Histologic lesions included splenic and hepatic necrosis associated with eosinophilic intranuclear inclusion bodies characteristic of herpesvirus. In one case, necrosis and inclusions were also noted in bone marrow, thymus, bursa of Fabricius, thyroid gland, parathyroid gland, ceca, and the enteric system. Transmission electron microscopy demonstrated viral particles typical of herpesvirus within hepatocyte nuclei and budding from the nuclear membrane. Herpesviral DNA was amplified via polymerase chain reaction (PCR) of paraffin-embedded liver and spleen, and sequence data were consistent with columbid herpesvirus-1, an alphaherpesvirus of Rock Pigeons (Columba livia). PCR results provide evidence that this disease is transmitted to raptors via Rock Pigeons, most likely through ingestion of Rock Pigeons as prey.

  10. Inclusion bodies induced by bean rugose mosaic virus seen under light microscopy

    Directory of Open Access Journals (Sweden)

    Carmen Rivera

    2000-12-01

    Full Text Available Two types of inclusion bodies were consistently observed under light microscopy in bean (Phaseolus vulgaris leaf tissue infected with bean rugose mosaic virus (BRMV, a species of the genus Comovirus, family Comoviridae. One type consisted of vacuolated inclusions found mainly in the cytoplasm of epidermal cells. The other type consisted of abundant crystalloid inclusions of different sizes and shapes found consistently in glandular hairs, guard cells, phloem tissue, xylem elements and occasionally in epidermal and mesophyll tissues. The two types of inclusion bodies stained with Azure A and Luxol Brilliant Green Bl-Calcomine Orange 2RS (O-G, and were similar to those seen to be caused by other species of comoviruses.Se observaron dos tipos de inclusiones virales, mediante microscopia de luz, en hojas de plantas de frijol (Phaseolus vulgaris previamente infectadas con el virus del mosaico rugoso del frijol ("bean rugose mosaic comovirus", BRMV, especie del género Comovirus, familia Comoviridae. Se hallaron inclusiones vesiculadas, principalmente en el citoplasma de células de la epidermis, y abundantes inclusiones cristalinas de diferentes formas y tamaños siempre en células guarda, tricomas glandulares, floema, elementos del xilema y ocasionalmente en células epidérmicas y del mesófilo. Ambos tipos de inclusiones tiñeron con Azure A y con la tinción, verde naranja (Luxol Brilliant Green BL-Calcomine Orange 2 RS conocida como OG, y son similares a las inclusiones inducidas por otras especies del género Comovirus.

  11. Inclusion body disease of falcons (Herpesvirus infection) in an American kestrel.

    Science.gov (United States)

    Kocan, A A; Potgieter, L N; Kocan, K M

    1977-04-01

    Postmortem examination of a captive-bred American kestrel (Falco sparverius) showed numerous white necrotic foci 1-2 mm in diameter throughout the liver and spleen. The results of light and spleen. The results of light and electron microscopic studies and experimental transmission to a captive American kestrel and a barred owl (Strix varia) suggests a herpesvirus similar to those dsecribed for owls and other falcons in the U.S. This is the first report of a naturally occurring case of inclusion body disease of falcons in the American kestrel.

  12. Infantile digital fibromatosis (inclusion body fibromatosis observed in a baby without finger involvement

    Directory of Open Access Journals (Sweden)

    Avni Kaya

    2013-01-01

    Full Text Available A 9-day-old male baby was hospitalized after his birth due to some swells under the skin. The hard consistency nodules observed under the skin all over the body of the patient were of different size, and presented lesions, among which the biggest was 1 × 1 cm. No lesions were observed on the fingers. By superficial ultrasonography, multiple isoechoic hypoechoic lesions were observed among the muscle plan. In thoracolumbar magnetic resonance imaging, multiple massif lesions retaining peripheral contrast (the biggest was 1.7 × 1.4 cm large had been observed under the skin muscle plans, between the muscles of the extremities. The biopsy was positive for smooth muscle actin, but negative for desmin, S100, and CD34. These findings were diagnosed as infantile digital fibromatosis (IDF (inclusion body fibromatosis. The case was presented with an objective to illustrate and remind that IDF can be observed in babies without finger involvement.

  13. Functional organization of cytoplasmic inclusion bodies in cells infected by respiratory syncytial virus.

    Science.gov (United States)

    Rincheval, Vincent; Lelek, Mickael; Gault, Elyanne; Bouillier, Camille; Sitterlin, Delphine; Blouquit-Laye, Sabine; Galloux, Marie; Zimmer, Christophe; Eleouet, Jean-François; Rameix-Welti, Marie-Anne

    2017-09-15

    Infection of cells by respiratory syncytial virus induces the formation of cytoplasmic inclusion bodies (IBs) where all the components of the viral RNA polymerase complex are concentrated. However, the exact organization and function of these IBs remain unclear. In this study, we use conventional and super-resolution imaging to dissect the internal structure of IBs. We observe that newly synthetized viral mRNA and the viral transcription anti-terminator M2-1 concentrate in IB sub-compartments, which we term "IB-associated granules" (IBAGs). In contrast, viral genomic RNA, the nucleoprotein, the L polymerase and its cofactor P are excluded from IBAGs. Live imaging reveals that IBAGs are highly dynamic structures. Our data show that IBs are the main site of viral RNA synthesis. They further suggest that shortly after synthesis in IBs, viral mRNAs and M2-1 transiently concentrate in IBAGs before reaching the cytosol and suggest a novel post-transcriptional function for M2-1.Respiratory syncytial virus (RSV) induces formation of inclusion bodies (IBs) sheltering viral RNA synthesis. Here, Rincheval et al. identify highly dynamic IB-associated granules (IBAGs) that accumulate newly synthetized viral mRNA and the viral M2-1 protein but exclude viral genomic RNA and RNA polymerase complexes.

  14. The 2-min walk test is sufficient for evaluating walking abilities in sporadic inclusion body myositis.

    Science.gov (United States)

    Alfano, L N; Lowes, L P; Dvorchik, I; Yin, H; Maus, E G; Flanigan, K M; Mendell, J R

    2014-03-01

    Sporadic inclusion body myositis causes progressive functional loss due to declining muscle strength. Although the underlying cause is unknown, clinical trials are underway to improve strength and function. Selection of appropriate outcome measures is critical for the success of these trials. The 6-min walk test has been the de facto standard for assessing function in neuromuscular disease; however, the optimal walking test has not been determined in this disease. In this study, 67 individuals with sporadic inclusion body myositis completed a battery of quantitative strength and functional tests including timed walking tests, patient-reported outcomes, and other tasks. The 2-min and 6-min walk tests are highly correlated to each other (r=0.97, pwalk test, but 7% of subjects were unable to walk the full 6-min of the 6-min walk test due to fatigue. The 2-min walk test demonstrates similar correlation to all outcomes compared to the 6-min walk test, is less fatiguing and better tolerated. Results suggest that the 2-min walk test is a better alternative to tests of longer duration. Further research is needed to determine longitudinal changes on this outcome. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. [A rare complication of dysarthria in a patient with inclusion body myositis: a case report].

    Science.gov (United States)

    Isayama, Reina; Shiga, Kensuke; Tanaka, Eijirou; Itsukage, Masahiro; Tokuda, Takahiko; Nakagawa, Masanori

    2010-10-01

    We reported a 71-year-old man with inclusion body myositis with clinically overt dysarthria. He had been suffering from gradual progression of weakness in the hand muscles and lower extremities as well as dysarthria three years before admission. His neurological examination revealed muscle atrophy and weakness in the tongue, the forearm flexors, and the vastus medialis muscles. He had dysarthria to a moderate degree, while he denied any dysphasia. A biopsy from vastus lateralis muscle showed variation in fiber size, infiltration of mononucleated cells, and numerous fibers with rimmed vacuoles, leading to the diagnosis of definite inclusion body myositis. The EMG findings of the tongue demonstrated low amplitude motor unit potentials during voluntary contraction, abundant fibrillation potentials at rest, and preserved interference pattern at maximal contraction, implying myogenic changes. We surmised the dysarthria seen in this patient, an atypical clinical feature in IBM, presumably caused by muscle involvement in the tongue muscle. Dysphasia is common symptom in IBM patient and has been much reported previously. But dysarthria in IBM patient has not been aware, for this reason this report should be the rare case.

  16. Gallium isotopic evidence for the fate of moderately volatile elements in planetary bodies and refractory inclusions

    Science.gov (United States)

    Kato, Chizu; Moynier, Frédéric

    2017-12-01

    The abundance of moderately volatile elements, such as Zn and Ga, show variable depletion relative to CI between the Earth and primitive meteorite (chondrites) parent bodies. Furthermore, the first solar system solids, the calcium-aluminum-rich inclusions (CAIs), are surprisingly rich in volatile element considering that they formed under high temperatures. Here, we report the Ga elemental and isotopic composition of a wide variety of chondrites along with five individual CAIs to understand the origin of the volatile elements and to further characterize the enrichment of the volatile elements in high temperature condensates. The δ71Ga (permil deviation of the 71Ga/69Ga ratio from the Ga IPGP standard) of carbonaceous chondrites decreases in the order of CI >CM >CO >CV and is inversely correlated with the Al/Ga ratio. This implies that the Ga budget of the carbonaceous chondrites parent bodies were inherited from a two component mixing of a volatile rich reservoir enriched in heavy isotope of Ga and a volatile poor reservoir enriched in light isotope of Ga. Calcium-aluminum-rich inclusions are enriched in Ga and Zn compared to the bulk meteorite and are both highly isotopically fractionated with δ71Ga down to -3.56‰ and δ66Zn down to -0.74‰. The large enrichment in the light isotopes of Ga and Zn in the CAIs implies that the moderately volatile elements were introduced in the CAIs during condensation in the solar nebula as opposed to secondary processing in the meteorite parent body and supports a change in gas composition in which CAIs were formed.

  17. A study of the cytoplasmic expression of a form of human prolactin and of its solubilization and renaturation from bacterial inclusion bodies; Estudo da expressao citoplasmatica bacteriana de uma forma de prolactina humana e de sua solubilizacao e renaturacao a partir de corpos de inclusao

    Energy Technology Data Exchange (ETDEWEB)

    Affonso, Regina

    2000-07-01

    Different vector elements, that can determine a high expression level of a form of human prolactin (taghPrl) in bacterial cytoplasm, were studied. Expression conditions were first optimized for a reference vector, which was used to transform different strains of E. coli: HB2151, RRI and RB791. The highest expression level (113 {+-}16 {mu}g/mL.A{sub 600}) was obtained in HB2151, after activation with only 0.1 mM IPTG. At this point the influence of the transcription terminator (g32 from bacteriophage T4), of the translation enhancer (g10 from bacteriophage T7), of the promoter ({lambda}P{sub L} or tac) and of the antibiotic resistance gene (amp{sup r} or kan{sup r}) were studied. The first three elements did not show any significant influence, at least in our systems. On the contrary, the analysis of the influence of amp{sup r} and kan{sup r} genes showed, unexpectedly, that the presence of the last one provides an approximately 5-fold higher expression for taghPrl in E. coli cytoplasm. Finally, an appropriate extraction, solubilization, renaturation and purification process, able to provide a monomeric form of taghPrl, was studied. A method utilizing urea and mercaptoethanol as solubilizing agents and a dialysis as a renaturation procedure, provided with some modifications, one of the highest yields ever reported in the literature: 35.4 {+-} 4.5% of total recovery. Moreover, the biological activity of the taghPrl obtained, when tested in the Nb2 cell proliferation assay, was of the same order of that shown by the International Standard of human prolactin of pituitary origin. These data show that the cytoplasmic expression system here described, which can provide an expression efficiency 50-100 - fold higher than the periplasmic expression, can represent a valid alternative for the production of this and of other hormones of pharmaceutical interest and grade. (author)

  18. Intracellular Bacterial Pathogens Trigger the Formation of U Small Nuclear RNA Bodies (U Bodies) through Metabolic Stress Induction.

    Science.gov (United States)

    Tsalikis, Jessica; Tattoli, Ivan; Ling, Arthur; Sorbara, Matthew T; Croitoru, David O; Philpott, Dana J; Girardin, Stephen E

    2015-08-21

    Invasive bacterial pathogens induce an amino acid starvation (AAS) response in infected host cells that controls host defense in part by promoting autophagy. However, whether AAS has additional significant effects on the host response to intracellular bacteria remains poorly characterized. Here we showed that Shigella, Salmonella, and Listeria interfere with spliceosomal U snRNA maturation in the cytosol. Bacterial infection resulted in the rerouting of U snRNAs and their cytoplasmic escort, the survival motor neuron (SMN) complex, to processing bodies, thus forming U snRNA bodies (U bodies). This process likely contributes to the decline in the cytosolic levels of U snRNAs and of the SMN complex proteins SMN and DDX20 that we observed in infected cells. U body formation was triggered by membrane damage in infected cells and was associated with the induction of metabolic stresses, such as AAS or endoplasmic reticulum stress. Mechanistically, targeting of U snRNAs to U bodies was regulated by translation initiation inhibition and the ATF4/ATF3 pathway, and U bodies rapidly disappeared upon removal of the stress, suggesting that their accumulation represented an adaptive response to metabolic stress. Importantly, this process likely contributed to shape the host response to invasive bacteria because down-regulation of DDX20 expression using short hairpin RNA (shRNA) amplified ATF3- and NF-κB-dependent signaling. Together, these results identify a critical role for metabolic stress and invasive bacterial pathogens in U body formation and suggest that this process contributes to host defense. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. Hepato nephropathology associated with inclusion body hepatitis complicated with citrinin mycotoxicosis in a broiler farm

    Directory of Open Access Journals (Sweden)

    Asok Kumar Mariappan

    2018-02-01

    Full Text Available Aim: Mortality in a broiler chicken farm was investigated for identifying the cause of mortality. Materials and Methods: A broiler farm with a population of 16000 succumbed to a disease outbreak. Clinical signs, vaccination history and mortality, were recorded. Necropsy examination and microscopic examination were carried out along with toxicological and molecular studies. Results: The clinical signs in the affected broiler birds were of non-specific nature with a total mortality of 26.39%. Postmortem examination and microscopical findings revealed hepatitis with basophilic intranuclear inclusion, splenitis, myocarditis, and nephritis. Glomerulonephritis was the prominent renal pathology recorded in this study. Polymerase chain reaction test confirmed the presence of fowl adenovirus (FAdV genome in the target organs, and toxicological examination by thin-layer chromatography revealed the presence of a toxic level of citrinin in the feed samples. Conclusion: Based on various diagnostic investigations, the mortality in the flock was attributed to inclusion body hepatitis (IBH complicated with citrinin mycotoxicosis. Thus, apart from liver pathology which occurs in a classical IBH cases, glomerulonephritis too occurs which are also a prominent finding which pathologists often miss. Thus, kidneys should also be examined histologically to assess the microscopic tissue alterations in poultry suspected for IBH along with a mycotoxicological analysis of feed. This will definitely throw light on the synergistic pathology elicited and exhibited by FAdV and mycotoxins in the poultry.

  20. Dynamic phosphorylation of Ebola virus VP30 in NP-induced inclusion bodies.

    Science.gov (United States)

    Lier, Clemens; Becker, Stephan; Biedenkopf, Nadine

    2017-12-01

    Zaire Ebolavirus (EBOV) causes a severe feverish disease with high case fatality rates. Transcription of EBOV is dependent on the activity of the nucleocapsid protein VP30 which represents an essential viral transcription factor. Activity of VP30 is regulated via phosphorylation at six N-terminal serine residues. Recent data demonstrated that dynamic phosphorylation and dephosphorylation of serine residue 29 is essential for transcriptional support activity of VP30. To analyze the spatio/temporal dynamics of VP30 phosphorylation, we generated a peptide antibody recognizing specifically VP30 phosphorylated at serine 29. Using this antibody we could demonstrate that (i) the majority of VP30 molecules in EBOV-infected cells is dephosphorylated at the crucial position serine 29, (ii) both, VP30 phosphorylation and dephosphorylation take place in viral inclusion bodies that are induced by the nucleoprotein NP and (iii) NP influences the phosphorylation state of VP30. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Efficient inclusion body processing using chemical extraction and high gradient magnetic fishing

    DEFF Research Database (Denmark)

    Heebøll-Nielsen, Anders; Choe, W.S.; Middelberg, A.P.J.

    2003-01-01

    In this study we introduce a radical new approach for the recovery of proteins expressed in the form of inclusion bodies, involving W chemical extraction from the host cells, (ii) adsorptive capture of the target protein onto small magnetic adsorbents, and (iii) subsequent rapid collection...... was achieved very rapidly ( 100 mM imidazole in the equilibration buffer. The influence of feedstock complexity on Ll adsorption to the Cu2+-charged type II magnetic chelators was studied using various dilutions of four crude chemical E. coli cell extracts containing denatured L I protein. Undiminished Ll...... adsorption to these adsorbents (relative to the 8 M urea-phosphate buffer case) was observed with the least complex of these feed materials, i.e., a partially clarified (12 g dry weight L-1) and spermine-treated chemical cell extract (feedstock B). Efficient recovery of Ll from feed B was demonstrated...

  2. Detection of novel divergent arenaviruses in boid snakes with inclusion body disease in The Netherlands.

    Science.gov (United States)

    Bodewes, R; Kik, M J L; Raj, V Stalin; Schapendonk, C M E; Haagmans, B L; Smits, S L; Osterhaus, A D M E

    2013-06-01

    Arenaviruses are bi-segmented negative-stranded RNA viruses, which were until recently only detected in rodents and humans. Now highly divergent arenaviruses have been identified in boid snakes with inclusion body disease (IBD). Here, we describe the identification of a new species and variants of the highly divergent arenaviruses, which were detected in tissues of captive boid snakes with IBD in The Netherlands by next-generation sequencing. Phylogenetic analysis of the complete sequence of the open reading frames of the four predicted proteins of one of the detected viruses revealed that this virus was most closely related to the recently identified Golden Gate virus, while considerable sequence differences were observed between the highly divergent arenaviruses detected in this study. These findings add to the recent identification of the highly divergent arenaviruses in boid snakes with IBD in the United States and indicate that these viruses also circulate among boid snakes in Europe.

  3. Efficient inclusion body processing using chemical extraction and high gradient magnetic fishing

    DEFF Research Database (Denmark)

    Heebøll-Nielsen, Anders; Choe, W.S.; Middelberg, A.P.J.

    2003-01-01

    was achieved very rapidly ( 100 mM imidazole in the equilibration buffer. The influence of feedstock complexity on Ll adsorption to the Cu2+-charged type II magnetic chelators was studied using various dilutions of four crude chemical E. coli cell extracts containing denatured L I protein. Undiminished Ll......In this study we introduce a radical new approach for the recovery of proteins expressed in the form of inclusion bodies, involving W chemical extraction from the host cells, (ii) adsorptive capture of the target protein onto small magnetic adsorbents, and (iii) subsequent rapid collection...... adsorption to these adsorbents (relative to the 8 M urea-phosphate buffer case) was observed with the least complex of these feed materials, i.e., a partially clarified (12 g dry weight L-1) and spermine-treated chemical cell extract (feedstock B). Efficient recovery of Ll from feed B was demonstrated...

  4. Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies.

    Science.gov (United States)

    Mačinković, Igor S; Abughren, Mohamed; Mrkic, Ivan; Grozdanović, Milica M; Prodanović, Radivoje; Gavrović-Jankulović, Marija

    2013-12-01

    High levels of recombinant protein expression can lead to the formation of insoluble inclusion bodies. These complex aggregates are commonly solubilized in strong denaturants, such as 6-8M urea, although, if possible, solubilization under milder conditions could facilitate subsequent refolding and purification of bioactive proteins. Commercially available GST-tag assays are designed for quantitative measurement of GST activity under native conditions. GST fusion proteins accumulated in inclusion bodies are considered to be undetectable by such assays. In this work, solubilization of recombinantly produced proteins was performed in 4M urea. The activity of rGST was assayed in 2M urea and it was shown that rGST preserves 85% of its activity under such denaturing conditions. A colorimetric GST activity assay with 1-chloro-2, 4-dinitrobenzene (CDNB) was examined for use in rapid detection of expression targeted to inclusion bodies and for the identification of inclusion body proteins which can be solubilized in low concentrations of chaotropic agents. Applicability of the assay was evaluated by tracking protein expression of two GST-fused allergens of biopharmaceutical value in E. coli, GST-Der p 2 and GST-Mus a 5, both targeted to inclusion bodies. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Positive in vitro wound healing effects of functional inclusion bodies of a lipoxygenase from the Mexican axolotl.

    Science.gov (United States)

    Stamm, Anne; Strauß, Sarah; Vogt, Peter; Scheper, Thomas; Pepelanova, Iliyana

    2018-04-07

    AmbLOXe is a lipoxygenase, which is up-regulated during limb-redevelopment in the Mexican axolotl, Ambystoma mexicanum, an animal with remarkable regeneration capacity. Previous studies have shown that mammalian cells transformed with the gene of this epidermal lipoxygenase display faster migration and wound closure rate during in vitro wound healing experiments. In this study, the gene of AmbLOXe was codon-optimized for expression in Escherichia coli and was produced in the insoluble fraction as protein aggregates. These inclusion bodies or nanopills were shown to be reservoirs containing functional protein during in vitro wound healing assays. For this purpose, functional inclusion bodies were used to coat cell culture surfaces prior cell seeding or were added directly to the medium after cells reached confluence. In both scenarios, AmbLOXe inclusion bodies led to faster migration rate and wound closure, in comparison to controls containing either no AmbLOXe or GFP inclusion bodies. Our results demonstrate that AmbLOXe inclusion bodies are functional and may serve as stable reservoirs of this enzyme. Nevertheless, further studies with soluble enzyme are also necessary in order to start elucidating the exact molecular substrates of AmbLOXe and the biochemical pathways involved in the wound healing effect.

  6. Thermal inclusions: how one spin can destroy a many-body localized phase

    Science.gov (United States)

    Ponte, Pedro; Laumann, C. R.; Huse, David A.; Chandran, A.

    2017-10-01

    Many-body localized (MBL) systems lie outside the framework of statistical mechanics, as they fail to equilibrate under their own quantum dynamics. Even basic features of MBL systems, such as their stability to thermal inclusions and the nature of the dynamical transition to thermalizing behaviour, remain poorly understood. We study a simple central spin model to address these questions: a two-level system interacting with strength J with N≫1 localized bits subject to random fields. On increasing J, the system transitions from an MBL to a delocalized phase on the vanishing scale Jc(N)˜1/N, up to logarithmic corrections. In the transition region, the single-site eigenstate entanglement entropies exhibit bimodal distributions, so that localized bits are either `on' (strongly entangled) or `off' (weakly entangled) in eigenstates. The clusters of `on' bits vary significantly between eigenstates of the same sample, which provides evidence for a heterogeneous discontinuous transition out of the localized phase in single-site observables. We obtain these results by perturbative mapping to bond percolation on the hypercube at small J and by numerical exact diagonalization of the full many-body system. Our results support the arguments that the MBL phase is unstable in systems with short-range interactions and quenched randomness in dimensions d that are high but finite. This article is part of the themed issue 'Breakdown of ergodicity in quantum systems: from solids to synthetic matter'.

  7. Enzymatic synthesis of c-di-GMP using inclusion bodies of Thermotoga maritima full-length diguanylate cyclase.

    Science.gov (United States)

    Korovashkina, A S; Rymko, A N; Kvach, S V; Zinchenko, A I

    2012-12-15

    Recombinant full-length diguanylate cyclases (DGCs) of Thermotoga maritima with native and mutant allosteric sites were overexpressed in Escherichia coli cells and characterized. It has been shown that target enzymes are produced substantially in the form of active inclusion bodies. Introduction of the mutation in allosteric site resulted in 7-fold increase of the T. maritima DGC activity. Possibility of applying full-length DGC of T. maritima in the form of inclusion bodies for synthesis of c-di-GMP was originally demonstrated. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. A novel and convenient method to immunize animals: Inclusion ...

    African Journals Online (AJOL)

    Purified recombinant protein has been routinely used to immunize rabbits to produce polyclonal antibodies. The process of the purification of recombinant proteins from bacterial inclusion bodies is both labor intensive and time consuming. To determine whether whole inclusion bodies can be used as antigen in polyclonal ...

  9. An efficient procedure for the expression and purification of HIV-1 protease from inclusion bodies.

    Science.gov (United States)

    Nguyen, Hong-Loan Thi; Nguyen, Thuy Thi; Vu, Quy Thi; Le, Hang Thi; Pham, Yen; Trinh, Phuong Le; Bui, Thuan Phuong; Phan, Tuan-Nghia

    2015-12-01

    Several studies have focused on HIV-1 protease for developing drugs for treating AIDS. Recombinant HIV-1 protease is used to screen new drugs from synthetic compounds or natural substances. However, large-scale expression and purification of this enzyme is difficult mainly because of its low expression and solubility. In this study, we constructed 9 recombinant plasmids containing a sequence encoding HIV-1 protease along with different fusion tags and examined the expression of the enzyme from these plasmids. Of the 9 plasmids, pET32a(+) plasmid containing the HIV-1 protease-encoding sequence along with sequences encoding an autocleavage site GTVSFNF at the N-terminus and TEV plus 6× His tag at the C-terminus showed the highest expression of the enzyme and was selected for further analysis. The recombinant protein was isolated from inclusion bodies by using 2 tandem Q- and Ni-Sepharose columns. SDS-PAGE of the obtained HIV-1 protease produced a single band of approximately 13 kDa. The enzyme was recovered efficiently (4 mg protein/L of cell culture) and had high specific activity of 1190 nmol min(-1) mg(-1) at an optimal pH of 4.7 and optimal temperature of 37 °C. This procedure for expressing and purifying HIV-1 protease is now being scaled up to produce the enzyme on a large scale for its application. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Capillary gel electrophoresis for the quantification and purity determination of recombinant proteins in inclusion bodies.

    Science.gov (United States)

    Espinosa-de la Garza, Carlos E; Perdomo-Abúndez, Francisco C; Campos-García, Víctor R; Pérez, Néstor O; Flores-Ortiz, Luis F; Medina-Rivero, Emilio

    2013-09-01

    In this work, a high-resolution CGE method for quantification and purity determination of recombinant proteins was developed, involving a single-component inclusion bodies (IBs) solubilization solution. Different recombinant proteins expressed as IBs were used to show method capabilities, using recombinant interferon-β 1b as the model protein for method validation. Method linearity was verified in the range from 0.05 to 0.40 mg/mL and a determination coefficient (r(2) ) of 0.99 was obtained. The LOQs and LODs were 0.018 and 0.006 mg/mL, respectively. RSD for protein content repeatability test was 2.29%. In addition, RSD for protein purity repeatability test was 4.24%. Method accuracy was higher than 90%. Specificity was confirmed, as the method was able to separate recombinant interferon-β 1b monomer from other aggregates and impurities. Sample content and purity was demonstrated to be stable for up to 48 h. Overall, this method is suitable for the analysis of recombinant proteins in IBs according to the attributes established on the International Conference for Harmonization guidelines. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Inclusion bodies as potential vehicles for recombinant protein delivery into epithelial cells

    Science.gov (United States)

    2012-01-01

    Background We present the potential of inclusion bodies (IBs) as a protein delivery method for polymeric filamentous proteins. We used as cell factory a strain of E. coli, a conventional host organism, and keratin 14 (K14) as an example of a complex protein. Keratins build the intermediate filament cytoskeleton of all epithelial cells. In order to build filaments, monomeric K14 needs first to dimerize with its binding partner (keratin 5, K5), which is then followed by heterodimer assembly into filaments. Results K14 IBs were electroporated into SW13 cells grown in culture together with a “reporter” plasmid containing EYFP labeled keratin 5 (K5) cDNA. As SW13 cells do not normally express keratins, and keratin filaments are built exclusively of keratin heterodimers (i.e. K5/K14), the short filamentous structures we obtained in this study can only be the result of: a) if both IBs and plasmid DNA are transfected simultaneously into the cell(s); b) once inside the cells, K14 protein is being released from IBs; c) released K14 is functional, able to form heterodimers with EYFP-K5. Conclusions Soluble IBs may be also developed for complex cytoskeletal proteins and used as nanoparticles for their delivery into epithelial cells. PMID:22624805

  12. Effects of solutes on solubilization and refolding of proteins from inclusion bodies with high hydrostatic pressure

    Science.gov (United States)

    Lee, Seung-Hyun; Carpenter, John F.; Chang, Byeong S.; Randolph, Theodore W.; Kim, Yong-Sung

    2006-01-01

    High hydrostatic pressure (HHP)-mediated solubilization and refolding of five inclusion bodies (IBs) produced from bacteria, three Gram-negative binding proteins (GNBP1, GNBP2, and GNBP3) from Drosophila, and two phosphatases from human were investigated in combination of a redox-shuffling agent (2 mM DTT and 6 mM GSSG) and various additives. HHP (200 MPa) combined with the redox-shuffling agent resulted in solubilization yields of ~42%–58% from 1 mg/mL of IBs. Addition of urea (1 and 2 M), 2.5 M glycerol, L-arginine (0.5 M), Tween 20 (0.1 mM), or Triton X-100 (0.5 mM) significantly enhanced the solubilization yield for all proteins. However, urea, glycerol, and nonionic surfactants populated more soluble oligomeric species than monomeric species, whereas arginine dominantly induced functional monomeric species (~70%–100%) to achieve refolding yields of ~55%–78% from IBs (1 mg/mL). Our results suggest that the combination of HHP with arginine is most effective in enhancing the refolding yield by preventing aggregation of partially folded intermediates populated during the refolding. Using the refolded proteins, the binding specificity of GNBP2 and GNBP3 was newly identified the same as with that of GNBP1, and the enzymatic activities of the two phosphatases facilitates their further characterization. PMID:16385003

  13. Anesthetic Considerations of Sporadic Inclusion Body Myositis in an Elderly Man With Orthopedic Trauma.

    Science.gov (United States)

    Steck, Dominik T; Choi, Christine; Gollapudy, Suneeta; Pagel, Paul S

    2016-04-01

    Sporadic inclusion body myositis (IBM) is an inflammatory myopathy characterized by progressive asymmetric extremity weakness, oropharyngeal dysphagia, and the potential for exaggerated sensitivity to neuromuscular blockers and respiratory compromise. The authors describe their management of a patient with IBM undergoing urgent orthopedic surgery. An 81-year-old man with IBM suffered a left intertrochanteric femoral fracture after falling down stairs. His IBM caused progressive left proximal lower extremity, bilateral distal upper extremity weakness (left > right), and oropharyngeal dysphagia (solid food, pills). He denied dyspnea, exercise intolerance, and a history of aspiration. Because respiratory insufficiency resulting from diaphragmatic dysfunction and prolonged duration of action of neuromuscular blockers may occur in IBM, the authors avoided using a neuromuscular blocker. After applying cricoid pressure, anesthesia was induced using intravenous lidocaine, propofol, remifentanil followed by manual ventilation with inhaled sevoflurane in oxygen. Endotracheal intubation was accomplished without difficulty; anesthesia was then maintained using remifentanil and sevoflurane. The fracture was repaired with a trochanteric femoral nail. The patient was extubated without difficulty and made an uneventful recovery. In summary, there is a lack of consensus about the use of neuromuscular blockers in patients with IBM. The authors avoided these drugs and were able to easily secure the patient's airway and maintain adequate muscle relaxation using a balanced sevoflurane-remifentanil anesthetic. Clinical trials are necessary to define the pharmacology of neuromuscular blockers in patients with IBM and determine whether use of these drugs contributes to postoperative respiratory insufficiency in these vulnerable patients.

  14. Purification of inclusion bodies using PEG precipitation under denaturing conditions to produce recombinant therapeutic proteins from Escherichia coli.

    Science.gov (United States)

    Chen, Huanhuan; Li, Ninghuan; Xie, Yueqing; Jiang, Hua; Yang, Xiaoyi; Cagliero, Cedric; Shi, Siwei; Zhu, Chencen; Luo, Han; Chen, Junsheng; Zhang, Lei; Zhao, Menglin; Feng, Lei; Lu, Huili; Zhu, Jianwei

    2017-07-01

    It has been documented that the purification of inclusion bodies from Escherichia coli by size exclusion chromatography (SEC) may benefit subsequent refolding and recovery of recombinant proteins. However, loading volume and the high cost of the column limits its application in large-scale manufacturing of biopharmaceutical proteins. We report a novel process using polyethylene glycol (PEG) precipitation under denaturing conditions to replace SEC for rapid purification of inclusion bodies containing recombinant therapeutic proteins. Using recombinant human interleukin 15 (rhIL-15) as an example, inclusion bodies of rhIL-15 were solubilized in 7 M guanidine hydrochloride, and rhIL-15 was precipitated by the addition of PEG 6000. A final concentration of 5% (w/v) PEG 6000 was found to be optimal to precipitate target proteins and enhance recovery and purity. Compared to the previously reported S-200 size exclusion purification method, PEG precipitation was easier to scale up and achieved the same protein yields and quality of the product. PEG precipitation also reduced manufacturing time by about 50 and 95% of material costs. After refolding and further purification, the rhIL-15 product was highly pure and demonstrated a comparable bioactivity with a rhIL-15 reference standard. Our studies demonstrated that PEG precipitation of inclusion bodies under denaturing conditions holds significant potential as a manufacturing process for biopharmaceuticals from E. coli protein expression systems.

  15. Dynamic JUNQ inclusion bodies are asymmetrically inherited in mammalian cell lines through the asymmetric partitioning of vimentin.

    Science.gov (United States)

    Ogrodnik, Mikołaj; Salmonowicz, Hanna; Brown, Rachel; Turkowska, Joanna; Średniawa, Władysław; Pattabiraman, Sundararaghavan; Amen, Triana; Abraham, Ayelet-chen; Eichler, Noam; Lyakhovetsky, Roman; Kaganovich, Daniel

    2014-06-03

    Aging is associated with the accumulation of several types of damage: in particular, damage to the proteome. Recent work points to a conserved replicative rejuvenation mechanism that works by preventing the inheritance of damaged and misfolded proteins by specific cells during division. Asymmetric inheritance of misfolded and aggregated proteins has been shown in bacteria and yeast, but relatively little evidence exists for a similar mechanism in mammalian cells. Here, we demonstrate, using long-term 4D imaging, that the vimentin intermediate filament establishes mitotic polarity in mammalian cell lines and mediates the asymmetric partitioning of damaged proteins. We show that mammalian JUNQ inclusion bodies containing soluble misfolded proteins are inherited asymmetrically, similarly to JUNQ quality-control inclusions observed in yeast. Mammalian IPOD-like inclusion bodies, meanwhile, are not always inherited by the same cell as the JUNQ. Our study suggests that the mammalian cytoskeleton and intermediate filaments provide the physical scaffold for asymmetric inheritance of dynamic quality-control JUNQ inclusions. Mammalian IPOD inclusions containing amyloidogenic proteins are not partitioned as effectively during mitosis as their counterparts in yeast. These findings provide a valuable mechanistic basis for studying the process of asymmetric inheritance in mammalian cells, including cells potentially undergoing polar divisions, such as differentiating stem cells and cancer cells.

  16. Application of an E. coli signal sequence as a versatile inclusion body tag.

    Science.gov (United States)

    Jong, Wouter S P; Vikström, David; Houben, Diane; van den Berg van Saparoea, H Bart; de Gier, Jan-Willem; Luirink, Joen

    2017-03-21

    Heterologous protein production in Escherichia coli often suffers from bottlenecks such as proteolytic degradation, complex purification procedures and toxicity towards the expression host. Production of proteins in an insoluble form in inclusion bodies (IBs) can alleviate these problems. Unfortunately, the propensity of heterologous proteins to form IBs is variable and difficult to predict. Hence, fusing the target protein to an aggregation prone polypeptide or IB-tag is a useful strategy to produce difficult-to-express proteins in an insoluble form. When screening for signal sequences that mediate optimal targeting of heterologous proteins to the periplasmic space of E. coli, we observed that fusion to the 39 amino acid signal sequence of E. coli TorA (ssTorA) did not promote targeting but rather directed high-level expression of the human proteins hEGF, Pla2 and IL-3 in IBs. Further analysis revealed that ssTorA even mediated IB formation of the highly soluble endogenous E. coli proteins TrxA and MBP. The ssTorA also induced aggregation when fused to the C-terminus of target proteins and appeared functional as IB-tag in E. coli K-12 as well as B strains. An additive effect on IB-formation was observed upon fusion of multiple ssTorA sequences in tandem, provoking almost complete aggregation of TrxA and MBP. The ssTorA-moiety was successfully used to produce the intrinsically unstable hEGF and the toxic fusion partner SymE, demonstrating its applicability as an IB-tag for difficult-to-express and toxic proteins. We present proof-of-concept for the use of ssTorA as a small, versatile tag for robust E. coli-based expression of heterologous proteins in IBs.

  17. Proteomics of rimmed vacuoles define new risk allele in inclusion body myositis.

    Science.gov (United States)

    Güttsches, Anne-Katrin; Brady, Stefen; Krause, Kathryn; Maerkens, Alexandra; Uszkoreit, Julian; Eisenacher, Martin; Schreiner, Anja; Galozzi, Sara; Mertens-Rill, Janine; Tegenthoff, Martin; Holton, Janice L; Harms, Matthew B; Lloyd, Thomas E; Vorgerd, Matthias; Weihl, Conrad C; Marcus, Katrin; Kley, Rudolf A

    2017-02-01

    Sporadic inclusion body myositis (sIBM) pathogenesis is unknown; however, rimmed vacuoles (RVs) are a constant feature. We propose to identify proteins that accumulate within RVs. RVs and intact myofibers were laser microdissected from skeletal muscle of 18 sIBM patients and analyzed by a sensitive mass spectrometry approach using label-free spectral count-based relative protein quantification. Whole exome sequencing was performed on 62 sIBM patients. Immunofluorescence was performed on patient and mouse skeletal muscle. A total of 213 proteins were enriched by >1.5 -fold in RVs compared to controls and included proteins previously reported to accumulate in sIBM tissue or when mutated cause myopathies with RVs. Proteins associated with protein folding and autophagy were the largest group represented. One autophagic adaptor protein not previously identified in sIBM was FYCO1. Rare missense coding FYCO1 variants were present in 11.3% of sIBM patients compared with 2.6% of controls (p = 0.003). FYCO1 colocalized at RVs with autophagic proteins such as MAP1LC3 and SQSTM1 in sIBM and other RV myopathies. One FYCO1 variant protein had reduced colocalization with MAP1LC3 when expressed in mouse muscle. This study used an unbiased proteomic approach to identify RV proteins in sIBM that included a novel protein involved in sIBM pathogenesis. FYCO1 accumulates at RVs, and rare missense variants in FYCO1 are overrepresented in sIBM patients. These FYCO1 variants may impair autophagic function, leading to RV formation in sIBM patient muscle. FYCO1 functionally connects autophagic and endocytic pathways, supporting the hypothesis that impaired endolysosomal degradation underlies the pathogenesis of sIBM. Ann Neurol 2017;81:227-239. © 2017 American Neurological Association.

  18. Intravenous immune globulin in hereditary inclusion body myopathy: a pilot study

    Directory of Open Access Journals (Sweden)

    Dorward Heidi

    2007-01-01

    Full Text Available Abstract Background Hereditary Inclusion Body Myopathy (HIBM is an autosomal recessive, adult onset, non-inflammatory neuromuscular disorder with no effective treatment. The causative gene, GNE, codes for UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase, which catalyzes the first two reactions in the synthesis of sialic acid. Reduced sialylation of muscle glycoproteins, such as α-dystroglycan and neural cell adhesion molecule (NCAM, has been reported in HIBM. Methods We treated 4 HIBM patients with intravenous immune globulin (IVIG, in order to provide sialic acid, because IgG contains 8 μmol of sialic acid/g. IVIG was infused as a loading dose of 1 g/kg on two consecutive days followed by 3 doses of 400 mg/kg at weekly intervals. Results For all four patients, mean quadriceps strength improved from 19.0 kg at baseline to 23.2 kg (+22% directly after IVIG loading to 25.6 kg (+35% at the end of the study. Mean shoulder strength improved from 4.1 kg at baseline to 5.9 kg (+44% directly after IVIG loading to 6.0 kg (+46% at the end of the study. The composite improvement for 8 other muscle groups was 5% after the initial loading and 19% by the end of the study. Esophageal motility and lingual strength improved in the patients with abnormal barium swallows. Objective measures of functional improvement gave variable results, but the patients experienced improvements in daily activities that they considered clinically significant. Immunohistochemical staining and immunoblotting of muscle biopsies for α-dystroglycan and NCAM did not provide consistent evidence for increased sialylation after IVIG treatment. Side effects were limited to transient headaches and vomiting. Conclusion The mild benefits in muscle strength experienced by HIBM patients after IVIG treatment may be related to the provision of sialic acid supplied by IVIG. Other sources of sialic acid are being explored as treatment options for HIBM.

  19. A variant of green fluorescent protein exclusively deposited to active intracellular inclusion bodies.

    Science.gov (United States)

    Raghunathan, Govindan; Munussami, Ganapathiraman; Moon, Hyojin; Paik, Hyun-jong; An, Seong Soo A; Kim, Yong-Sung; Kang, Sebyung; Lee, Sun-Gu

    2014-05-16

    Inclusion bodies (IBs) were generally considered to be inactive protein deposits and did not hold any attractive values in biotechnological applications. Recently, some IBs of recombinant proteins were confirmed to show their functional properties such as enzyme activities, fluorescence, etc. Such biologically active IBs are not commonly formed, but they have great potentials in the fields of biocatalysis, material science and nanotechnology. In this study, we characterized the IBs of DL4, a deletion variant of green fluorescent protein which forms active intracellular aggregates. The DL4 proteins expressed in Escherichia coli were exclusively deposited to IBs, and the IBs were estimated to be mostly composed of active proteins. The spectral properties and quantum yield of the DL4 variant in the active IBs were almost same with those of its native protein. Refolding and stability studies revealed that the deletion mutation in DL4 didn't affect the folding efficiency of the protein, but destabilized its structure. Analyses specific for amyloid-like structures informed that the inner architecture of DL4 IBs might be amorphous rather than well-organized. The diameter of fluorescent DL4 IBs could be decreased up to 100-200 nm by reducing the expression time of the protein in vivo. To our knowledge, DL4 is the first GFP variant that folds correctly but aggregates exclusively in vivo without any self-aggregating/assembling tags. The fluorescent DL4 IBs have potentials to be used as fluorescent biomaterials. This study also suggests that biologically active IBs can be achieved through engineering a target protein itself.

  20. Avian Adenoviruses Infections with Special Attention to Inclusion Body Hepatitis/ Hydropericardium Syndrome and Egg Drop Syndrome

    Directory of Open Access Journals (Sweden)

    Hafez Mohamed Hafez*

    2011-04-01

    Full Text Available The first avian adenovirus (AAV associated with clinical disease was isolated from an outbreak of respiratory disease in quail in 1950 (Olson, 1950. Since that time, AAVs have been found in all types and breeds of chickens and from a variety of other avian species. The infections may be asymptomatic or associated with several clinical and pathological conditions. Vertical transmission via the egg is the most common way of transmission. Also horizontal transmission through faeces, contaminated egg trays, crates and trucks play a role in the infection route. Studies have demonstrated the presence of antibodies in healthy poultry, and viruses have been isolated from normal birds. Avian adenoviruses in chickens are the etiological agents of 2 diseases known as inclusion body hepatitis (IBH and hydropericardium syndrome (HP. In some cases each condition is observed separately, however, recently the 2 conditions have frequently been observed as a single entity; therefore, the name hepatitis hydropericardium has been widely used to describe the pathologic condition. The syndrome is an acute disease of young chickens associated with anemia, haemorrhagic disorders, hydropericardium and high mortality. Egg-Drop-Syndrome (EDS is caused also by an adenovirus. The disease is characterised by a severe drop in egg production as well as the production of shell-less, thin-shelled, discoloured or misshapen eggs in apparently healthy birds. Ducks and geese are the natural host of the EDS virus. It was first described in chickens in the 1970s and spread to several countries world wide. The birds usually do not show any other signs of disease, and mortality is not expected. There is no specific treatment of the AAV infections. Active immunization by vaccination using an inactivated is wide spread.

  1. Resistance training with vascular occlusion in inclusion body myositis: a case study.

    Science.gov (United States)

    Gualano, Bruno; Neves, Manoel; Lima, Fernanda Rodrigues; Pinto, Ana Lúcia De Sá; Laurentino, Gilberto; Borges, Claudia; Baptista, Luciana; Artioli, Guilherme Giannini; Aoki, Marcelo Saldanha; Moriscot, Anselmo; Lancha, Antonio Herbert; Bonfá, Eloísa; Ugrinowitsch, Carlos

    2010-02-01

    Inclusion body myositis (IBM) is a rare idiopathic inflammatory myopathy that produces remarkable muscle weakness. Resistance training with vascular occlusion has been shown to improve muscle strength and cross-sectional area in other muscle wasting conditions. We evaluated the efficacy of a moderate-intensity resistance training program combined with vascular occlusion by examining functional capacity, muscle morphology, and changes in the expression of genes related to muscle protein synthesis and proteolysis in a patient with IBM. A 65-yr-old man with IBM resistant to all proposed treatments underwent resistance training with vascular occlusion for 12 wk. Leg press one-repetition maximum; thigh cross-sectional area; balance, mobility, and muscle function; quality of life; and blood markers of inflammation and muscle damage were assessed at baseline and after the 12-wk program. The messenger RNA (mRNA) expression levels of mechanogrowth factor, mammalian target of rapamycin, atrogin-1, and muscle RING finger-1 were also quantified. After the 12-wk training program, the patient's leg press one-repetition maximum, balance and mobility function, and thigh cross-sectional area increased 15.9%, 60%, and 4.7%, respectively. All Short Form-36 Health Survey Questionnaire subscales demonstrated improvements as well, varying from 18% to 600%. mRNA expression of mechanogrowth factor increased 3.97-fold, whereas that of atrogin-1 decreased 0.62-fold. Muscle RING finger-1 and mammalian target of rapamycin mRNA levels were only slightly altered, 1.18- and 1.28-fold, respectively. Importantly, the exercise did not induce disease flare. We describe a novel, and likely the first, nonpharmacological therapeutic tool that might be able to counteract the muscle atrophy and the declining strength that usually occur in IBM.

  2. Evaluation of the inclusion of circular RNAs in mRNA profiling in forensic body fluid identification.

    Science.gov (United States)

    Zhang, Yaqi; Liu, Baonian; Shao, Chengchen; Xu, Hongmei; Xue, Aimin; Zhao, Ziqin; Shen, Yiwen; Tang, Qiqun; Xie, Jianhui

    2018-01-01

    The use of messenger RNA (mRNA) profiling is considered a promising method in the identification of forensically relevant body fluids which can provide crucial information for reconstructing a potential crime. However, casework samples are usually of limited quantity or have been subjected to degradation, which requires improvement of body fluid identification. Circular RNAs (circRNAs), a class of products from the backsplicing of pre-mRNAs, are shown to have high abundance, remarkable stability, and cell type-specific expression in human cells. In this study, we investigated whether the inclusion of circRNAs in mRNA profiling improve the detection of biomarkers including δ-aminolevulinate synthase 2 (ALAS2) and matrix metallopeptidase 7 (MMP7) in body fluid identification. The major circRNAs of ALAS2 and MMP7 were first identified and primer sets for the simultaneous detection of linear and circular transcripts were developed. The inclusion of circRNAs in mRNA profiling showed improved detection sensitivity and stability of biomarkers revealed by using serial dilutions, mixed samples, and menstrual bloodstains as well as degraded and aged samples. Therefore, the inclusion of circRNAs in mRNA profiling should facilitate the detection of mRNA markers in forensic body fluid identification.

  3. Cooperation between two ClpB isoforms enhances the recovery of the recombinant {beta}-galactosidase from inclusion bodies

    Energy Technology Data Exchange (ETDEWEB)

    Guenther, Izabela [Department of Biochemistry, University of Gdansk, Wita Stwosza 59, 80-308 Gdansk (Poland); Zolkiewski, Michal [Department of Biochemistry, Kansas State University, Manhattan, KS 66506 (United States); Kedzierska-Mieszkowska, Sabina, E-mail: kedzie@biotech.ug.gda.pl [Department of Biochemistry, University of Gdansk, Wita Stwosza 59, 80-308 Gdansk (Poland)

    2012-10-05

    Highlights: Black-Right-Pointing-Pointer An important role of synergistic cooperation between the two ClpB isoforms. Black-Right-Pointing-Pointer Both ClpB isoforms are associated with IBs of {beta}-galactosidase. Black-Right-Pointing-Pointer ClpB is a key chaperone in IB protein release. -- Abstract: Bacterial ClpB is a molecular chaperone that solubilizes and reactivates aggregated proteins in cooperation with the DnaK chaperone system. The mechanism of protein disaggregation mediated by ClpB is linked to translocation of substrates through the central channel within the ring-hexameric structure of ClpB. Two isoforms of ClpB are produced in vivo: the full-length ClpB95 and the truncated ClpB80 (ClpB{Delta}N), which does not contain the N-terminal domain. The functional specificity of the two ClpB isoforms and the biological role of the N-terminal domain are still not fully understood. Recently, it has been demonstrated that ClpB may achieve its full potential as an aggregate-reactivating chaperone through the functional interaction and synergistic cooperation of its two isoforms. It has been found that the most efficient resolubilization and reactivation of stress-aggregated proteins occurred in the presence of both ClpB95 and ClpB80. In this work, we asked if the two ClpB isoforms functionally cooperate in the solubilization and reactivation of proteins from insoluble inclusion bodies (IBs) in Escherichia coli cells. Using the model {beta}-galactosidase fusion protein (VP1LAC), we found that solubilization and reactivation of enzymes entrapped in IBs occurred more efficiently in the presence of ClpB95 with ClpB80 than with either ClpB95 or ClpB80 alone. The two isoforms of ClpB chaperone acting together enhanced the solubility and enzymatic activity of {beta}-galactosidase sequestered into IBs. Both ClpB isoforms were associated with IBs of {beta}-galactosidase, what demonstrates their affinity to this type of aggregates. These results demonstrate a synergistic

  4. Cooperation between two ClpB isoforms enhances the recovery of the recombinant β-galactosidase from inclusion bodies

    International Nuclear Information System (INIS)

    Guenther, Izabela; Zolkiewski, Michal; Kędzierska-Mieszkowska, Sabina

    2012-01-01

    Highlights: ► An important role of synergistic cooperation between the two ClpB isoforms. ► Both ClpB isoforms are associated with IBs of β-galactosidase. ► ClpB is a key chaperone in IB protein release. -- Abstract: Bacterial ClpB is a molecular chaperone that solubilizes and reactivates aggregated proteins in cooperation with the DnaK chaperone system. The mechanism of protein disaggregation mediated by ClpB is linked to translocation of substrates through the central channel within the ring-hexameric structure of ClpB. Two isoforms of ClpB are produced in vivo: the full-length ClpB95 and the truncated ClpB80 (ClpBΔN), which does not contain the N-terminal domain. The functional specificity of the two ClpB isoforms and the biological role of the N-terminal domain are still not fully understood. Recently, it has been demonstrated that ClpB may achieve its full potential as an aggregate-reactivating chaperone through the functional interaction and synergistic cooperation of its two isoforms. It has been found that the most efficient resolubilization and reactivation of stress-aggregated proteins occurred in the presence of both ClpB95 and ClpB80. In this work, we asked if the two ClpB isoforms functionally cooperate in the solubilization and reactivation of proteins from insoluble inclusion bodies (IBs) in Escherichia coli cells. Using the model β-galactosidase fusion protein (VP1LAC), we found that solubilization and reactivation of enzymes entrapped in IBs occurred more efficiently in the presence of ClpB95 with ClpB80 than with either ClpB95 or ClpB80 alone. The two isoforms of ClpB chaperone acting together enhanced the solubility and enzymatic activity of β-galactosidase sequestered into IBs. Both ClpB isoforms were associated with IBs of β-galactosidase, what demonstrates their affinity to this type of aggregates. These results demonstrate a synergistic cooperation between the two isoforms of ClpB chaperone. In addition, no significant recovery of

  5. Formation of active inclusion bodies induced by hydrophobic self-assembling peptide GFIL8.

    Science.gov (United States)

    Wang, Xu; Zhou, Bihong; Hu, Weike; Zhao, Qing; Lin, Zhanglin

    2015-06-16

    In the last few decades, several groups have observed that proteins expressed as inclusion bodies (IBs) in bacteria could still be biologically active when terminally fused to an appropriate aggregation-prone partner such as pyruvate oxidase from Paenibacillus polymyxa (PoxB). More recently, we have demonstrated that three amphipathic self-assembling peptides, an alpha helical peptide 18A, a beta-strand peptide ELK16, and a surfactant-like peptide L6KD, have properties that induce target proteins into active IBs. We have developed an efficient protein expression and purification approach for these active IBs by introducing a self-cleavable intein molecule. In this study, the self-assembling peptide GFIL8 (GFILGFIL) with only hydrophobic residues was analyzed, and this peptide effectively induced the formation of cytoplasmic IBs in Escherichia coli when terminally attached to lipase A and amadoriase II. The protein aggregates in cells were confirmed by transmission electron microscopy analysis and retained ~50% of their specific activities relative to the native counterparts. We constructed an expression and separation coupled tag (ESCT) by incorporating an intein molecule, the Mxe GyrA intein. Soluble target proteins were successfully released from active IBs upon cleavage of the intein between the GFIL8 tag and the target protein, which was mediated by dithiothreitol. A variant of GFIL8, GFIL16 (GFILGFILGFILGFIL), improved the ESCT scheme by efficiently eliminating interference from the soluble intein-GFIL8 molecule. The yields of target proteins at the laboratory scale were 3.0-7.5 μg/mg wet cell pellet, which is comparable to the yields from similar ESCT constructs using 18A, ELK16, or the elastin-like peptide tag scheme. The all-hydrophobic self-assembling peptide GFIL8 induced the formation of active IBs in E. coli when terminally attached to target proteins. GFIL8 and its variant GFIL16 can act as a "pull-down" tag to produce purified soluble proteins with

  6. Expression of Mycobacterium tuberculosis Protein Tyrosine Phosphatase B in Escherichia coli and Its Recovery from Inclusion Body

    Directory of Open Access Journals (Sweden)

    Lalu Rudyat Telly Savalas

    2017-12-01

    Full Text Available The present study aims at expressing and partially purifying PtpB in active form. To achieve this, Mtb PtpB gene has been cloned into pET30a vector and overexpressed in Escherichia coli BL 21(DE3 under IPTG induction in the form of an inclusion body. Following resolubilization by urea and dialysis, the resulted PtpB has been shown to be active against para-Nitrophenyl phosphate.  It is concluded that the resulted PtpB has had been recovered from inclusion body to give the active form of the enzyme, and thus the success in overexpressing PtpB provides the required material to investigate the biochemical properties of the pathogen virulence factor further. 

  7. Anti-aggregatory effect of cyclodextrins in the refolding process of recombinant growth hormones from Escherichia coli inclusion bodies

    DEFF Research Database (Denmark)

    Bajorunaite, Egle; Cirkovas, Andrejus; Radzevicius, Kostas

    2009-01-01

    Cyclodextrins with different ring size and ring substituents were tested for recombinant mink and porcine growth hormones aggregation suppression in the refolding process from Escherichia coli inclusion bodies. Methyl-β-cyclodextrin and 2-hydroxypropyl-β-cyclodextrin show a positive effect on the......-β-cyclodextrin concentration. In conclusion, cyclodextrins have perspectives in biotechnology of veterinary growth hormones not only for protein production, but also for its storage......Cyclodextrins with different ring size and ring substituents were tested for recombinant mink and porcine growth hormones aggregation suppression in the refolding process from Escherichia coli inclusion bodies. Methyl-β-cyclodextrin and 2-hydroxypropyl-β-cyclodextrin show a positive effect...... on the aggregation suppression of both proteins. The influence of different methyl-β-cyclodextrin and 2-hydroxypropyl-β-cyclodextrin concentrations on the renaturation yield of both growth hormones was investigated. Moreover, methyl-β-cyclodextrin and 2-hydroxypropyl-β-cyclodextrin suppress not only folding...

  8. Access to gram scale amounts of functional globular adiponectin from E. coli inclusion bodies by alkaline-shock solubilization.

    Science.gov (United States)

    Heiker, John T; Klöting, Nora; Blüher, Matthias; Beck-Sickinger, Annette G

    2010-07-16

    The adipose tissue derived protein adiponectin exerts anti-diabetic, anti-inflammatory and anti-atherosclerotic effects. Adiponectin serum concentrations are in the microgram per milliliter range in healthy humans and inversely correlate with obesity and metabolic disorders. Accordingly, raising circulating adiponectin levels by direct administration may be an intriguing strategy in the treatment of obesity-related metabolic disorders. However production of large amounts of recombinant adiponectin protein is a primary obstacle so far. Here, we report a novel method for large amount production of globular adiponectin from E. coli inclusion bodies utilizing an alkaline-shock solubilization method without chaotropic agents followed by precipitation of the readily renaturing protein. Precipitation of the mildly solubilized protein capitalizes on advantages of inclusion body formation. This approach of inclusion body protein recovery provides access to gram scale amounts of globular adiponectin with standard laboratory equipment avoiding vast dilution or dialysis steps to neutralize the pH and renature the protein, thus saving chemicals and time. The precipitated protein is readily renaturing in buffer, is of adequate purity without a chromatography step and shows biological activity in cultured MCF7 cells and significantly lowered blood glucose levels in mice with streptozotocin induced type 1 diabetes. Copyright 2010 Elsevier Inc. All rights reserved.

  9. On the reconstruction of inclusions in a heat conductive body from dynamical boundary data over a finite time interval

    International Nuclear Information System (INIS)

    Ikehata, Masaru; Kawashita, Mishio

    2010-01-01

    The enclosure method was originally introduced for inverse problems concerning non-destructive evaluation governed by elliptic equations. It was developed as one of the useful approaches in inverse problems and applied for various equations. In this paper, an application of the enclosure method to an inverse initial boundary value problem for a parabolic equation with a discontinuous coefficient is given. A simple method to extract the depth of unknown inclusions in a heat conductive body from a single set of the temperature and heat flux on the boundary observed over a finite time interval is introduced. Other related results with infinitely many data are also reported. One of them gives the minimum radius of the open ball centred at a given point that contains the inclusions. The formula for the minimum radius is newly discovered

  10. Mechanical Ventilation Weaning in Inclusion Body Myositis: Feasibility of Isokinetic Inspiratory Muscle Training as an Adjunct Therapy

    Directory of Open Access Journals (Sweden)

    Leonardo Cordeiro de Souza

    2014-01-01

    Full Text Available Inclusion body myositis is a rare myopathy associated with a high rate of respiratory complications. This condition usually requires prolonged mechanical ventilation and prolonged intensive care stay. The unsuccessful weaning is mainly related to respiratory muscle weakness that does not promptly respond to immunosuppressive therapy. We are reporting a case of a patient in whom the use of an inspiratory muscle-training program which started after a two-week period of mechanical ventilation was associated with a successful weaning in one week and hospital discharge after 2 subsequent weeks.

  11. Injection of Botulinum Toxin a to Upper Esophageal Sphincter for Oropharyngeal Dysphagia in Two Patients with Inclusion Body Myositis

    Directory of Open Access Journals (Sweden)

    Louis WC Liu

    2004-01-01

    Full Text Available Inclusion body myositis (IBM is a progressive degenerative skeletal muscle disease leading to weakening and atrophy of both proximal and distal muscles. Dysphagia is reported in up to 86% of IBM patients. Surgical cricopharyngeal myotomy may be effective for cricopharyngeal dysphagia and there is one published report that botulinum toxin A, injected into the cricopharyngeus muscle using a hypopharyngoscope under general anesthesia, relieved IBM-associated dysphagia. This report presents the first documentation of botulinum toxin A injection into the upper esophageal sphincter using a flexible esophagogastroduodenoscope under conscious sedation, to reduce upper esophageal sphincter pressure and successfully alleviate oropharyngeal dysphagia in two IBM patients.

  12. No mutations in hnRNPA1 and hnRNPA2B1 in Dutch patients with amyotrophic lateral sclerosis, frontotemporal dementia, and inclusion body myopathy

    NARCIS (Netherlands)

    Seelen, Meinie; Visser, Anne E.; Overste, Daniel J.; Kim, Hong J.; Palud, A.; Wong, Tsz H.; van Swieten, John C.; Scheltens, Philip; Voermans, Nicol C.; Baas, Frank; de Jong, J. M. B. V.; van der Kooi, Anneke J.; de Visser, Marianne; Veldink, Jan H.; Taylor, J. Paul; van Es, Michael A.; van den Berg, Leonard H.

    2014-01-01

    Inclusion body myopathy (IBM) associated with Paget disease of the bone, frontotemporal dementia (FTD), and amyotrophic lateral sclerosis (ALS), sometimes called IBMPFD/ALS or multi system proteinopathy, is a rare, autosomal dominant disorder characterized by progressive degeneration of muscle,

  13. The bacterial communities associated with fecal types and body weight of rex rabbits.

    Science.gov (United States)

    Zeng, Bo; Han, Shushu; Wang, Ping; Wen, Bin; Jian, Wensu; Guo, Wei; Yu, Zhiju; Du, Dan; Fu, Xiangchao; Kong, Fanli; Yang, Mingyao; Si, Xiaohui; Zhao, Jiangchao; Li, Ying

    2015-03-20

    Rex rabbit is an important small herbivore for fur and meat production. However, little is known about the gut microbiota in rex rabbit, especially regarding their relationship with different fecal types and growth of the hosts. We characterized the microbiota of both hard and soft feces from rex rabbits with high and low body weight by using the Illumina MiSeq platform targeting the V4 region of the 16S rDNA. High weight rex rabbits possess distinctive microbiota in hard feces, but not in soft feces, from the low weight group. We detected the overrepresentation of several genera such as YS2/Cyanobacteria, and Bacteroidales and underrepresentation of genera such as Anaeroplasma spp. and Clostridiaceae in high weight hard feces. Between fecal types, several bacterial taxa such as Ruminococcaceae, and Akkermansia spp. were enriched in soft feces. PICRUSt analysis revealed that metabolic pathways such as "stilbenoid, diarylheptanoid, gingerol biosynthesis" were enriched in high weight rabbits, and pathways related to "xenobiotics biodegradation" and "various types of N-glycan biosynthesis" were overrepresented in rabbit soft feces. Our study provides foundation to generate hypothesis aiming to test the roles that different bacterial taxa play in the growth and caecotrophy of rex rabbits.

  14. High and compact formation of baculoviral polyhedrin-induced inclusion body by co-expression of baculoviral FP25 in Escherichia coli.

    Science.gov (United States)

    Li, Lin; Kim, Young Soo; Hwang, Dong Soo; Seo, Jeong Hyun; Jung, Hee Jung; Du, Juan; Cha, Hyung Joon

    2007-04-15

    Previously, we found that baculoviral polyhedrin (Polh) can successfully be used in Escherichia coli as a fusion partner for the expression of special foreign proteins as inclusion bodies, and the resulting, easily isolatable Polh-induced fusion inclusion bodies had almost the same characteristics as the native Polh. Here, we investigated the effects of co-expression of baculoviral FP25 protein on Polh-induced inclusion-body production in an E. coli expression system, as FP25 is known to be involved specifically in polyhedra formation. Using several analytical tools, including SDS-PAGE, pronase proteolysis, solubilization under alkaline conditions, and electron microscopy, we found that co-expressed FP25 was associated with Polh-induced inclusion bodies and that its co-expression led to formation of compact inclusion bodies as well as high production levels. We confirmed that FP25 co-expression induced higher production levels of other heterologous protein, antimicrobial peptide Hal18, fused with aggregation-prone Polh. Therefore, co-expression of baculoviral FP25 can be promisingly used to increase the levels of baculoviral Polh-fused foreign proteins, especially harmful proteins, expressed as inclusion bodies in an E. coli expression system. (c) 2006 Wiley Periodicals, Inc.

  15. Neutral weak-current two-body contributions in inclusive scattering from {sup 12}C

    Energy Technology Data Exchange (ETDEWEB)

    Lovato, Alessandro [ANL; Gandolfi, Stefano [LANL; Carlson, Joseph [LANL; Pieper, S. C. [ANL; Schiavilla, Rocco [JLAB, ODU

    2014-05-01

    An {\\it ab initio} calculation of the sum rules of the neutral weak response functions in $^{12}$C is reported, based on a realistic Hamiltonian, including two- and three-nucleon potentials, and on realistic currents, consisting of one- and two-body terms. We find that the sum rules of the response functions associated with the longitudinal and transverse components of the (space-like) neutral current are largest and that a significant portion ($\\simeq 30$\\%) of the calculated strength is due to two-body terms. This fact may have implications for the MiniBooNE and other neutrino quasi-elastic scattering data on nuclei.

  16. Effects of Inclusion Levels of Wheat Bran and Body Weight on Ileal and Fecal Digestibility in Growing Pigs

    Directory of Open Access Journals (Sweden)

    Q. Huang

    2015-06-01

    Full Text Available The objective of this study was to determine the effects of graded inclusions of wheat bran (0%, 9.65%, 48.25% wheat bran and two growth stages (from 32.5 to 47.2 kg and 59.4 to 78.7 kg, respectively on the apparent ileal digestibility (AID, apparent total tract digestibility (ATTD and hindgut fermentation of nutrients and energy in growing pigs. Six light pigs (initial body weight [BW] 32.5±2.1 kg and six heavy pigs (initial BW 59.4±3.2 kg were surgically prepared with a T-cannula in the distal ileum. A difference method was used to calculate the nutrient and energy digestibility of wheat bran by means of comparison with a basal diet consisting of corn-soybean meal (0% wheat bran. Two additional diets were formulated by replacing 9.65% and 48.25% wheat bran by the basal diet, respectively. Each group of pigs was allotted to a 6×3 Youden square design, and pigs were fed to three experimental diets during three 11-d periods. Hindgut fermentation values were calculated as the differences between ATTD and AID values. For the wheat bran diets, the AID and ATTD of dry matter (DM, ash, organic matter (OM, carbohydrates (CHO, gross energy (GE, and digestible energy (DE decreased with increasing inclusion levels of wheat bran (p<0.05. While only AID of CHO and ATTD of DM, ash, OM, CHO, GE, and DE content differed (p<0.05 when considering the BW effect. For the wheat bran ingredient, there was a wider variation effect (p<0.01 on the nutrient and energy digestibility of wheat bran in 9.65% inclusion level due to the coefficient of variation (CV of the nutrient and energy digestibility being higher at 9.65% compared to 48.25% inclusion level of wheat bran. Digestible energy content of wheat bran at 48.25% inclusion level (4.8 and 6.7 MJ/kg of DM, respectively fermented by hindgut was significantly higher (p<0.05 than that in 9.65% wheat bran inclusion level (2.56 and 2.12 MJ/kg of DM, respectively, which was also affected (p<0.05 by two growth stages

  17. Data on enhanced expression and purification of camelid single domain antibodies from Escherichia coli classical inclusion bodies.

    Science.gov (United States)

    Maggi, Maristella; Scotti, Claudia

    2017-06-01

    Heterologous expression of high amounts of recombinant proteins is a milestone for research and industrial purposes. Single domain antibodies (sdAbs) are heavy-chain only antibody fragments with applications in the biotechnological, medical and industrial fields. The simple nature and small size of sdAbs allows for efficient expression of the soluble molecule in different hosts. However, in some cases, it results in low functional protein yield. To overcome this limitation, expression of a 6xHistag sdAb was attempted in different conditions in Escherichia coli BL21(DE3) cells. Data showed that high amount of sdAb can be expressed in E. coli classical inclusion bodies, efficiently extracted by urea in a short-time, and properly purified by metal ion affinity chromatography. These data originate from the research article "Enhanced expression and purification of camelid single domain VHH antibodies from classical inclusion bodies" Maggi and Scotti (2017) [1] (DOI: http://dx.doi.org/10.1016/j.pep.2017.02.007).

  18. Purification and Refolding to Amyloid Fibrils of (His)6-tagged Recombinant Shadoo Protein Expressed as Inclusion Bodies in E. coli.

    Science.gov (United States)

    Li, Qiaojing; Richard, Charles-Adrien; Moudjou, Mohammed; Vidic, Jasmina

    2015-12-19

    The Escherichia coli expression system is a powerful tool for the production of recombinant eukaryotic proteins. We use it to produce Shadoo, a protein belonging to the prion family. A chromatographic method for the purification of (His)6-tagged recombinant Shadoo expressed as inclusion bodies is described. The inclusion bodies are solubilized in 8 M urea and bound to a Ni(2+)-charged column to perform ion affinity chromatography. Bound proteins are eluted by a gradient of imidazole. Fractions containing Shadoo protein are subjected to size exclusion chromatography to obtain a highly purified protein. In the final step purified Shadoo is desalted to remove salts, urea and imidazole. Recombinant Shadoo protein is an important reagent for biophysical and biochemical studies of protein conformation disorders occurring in prion diseases. Many reports demonstrated that prion neurodegenerative diseases originate from the deposition of stable, ordered amyloid fibrils. Sample protocols describing how to fibrillate Shadoo into amyloid fibrils at acidic and neutral/basic pHs are presented. The methods on how to produce and fibrillate Shadoo can facilitate research in laboratories working on prion diseases, since it allows for production of large amounts of protein in a rapid and low cost manner.

  19. Creating Inclusive Physical Activity Spaces: The Case of Body-Positive Yoga

    Science.gov (United States)

    Pickett, Andrew C.; Cunningham, George B.

    2017-01-01

    Purpose: Within the modern cultural climate, those in larger bodies face high levels of weight stigma, particularly in sport and physical activity spaces, which serves as a strong barrier to their participation. However, given the strong link between physical activity and general health and well-being for participants, it is important to explore…

  20. Could bacteriophages transfer antibiotic resistance genes from environmental bacteria to human-body associated bacterial populations?

    Science.gov (United States)

    Muniesa, Maite; Colomer-Lluch, Marta; Jofre, Juan

    2013-07-01

    Environments without any contact with anthropogenic antibiotics show a great abundance of antibiotic resistance genes that use to be chromosomal and are part of the core genes of the species that harbor them. Some of these genes are shared with human pathogens where they appear in mobile genetic elements. Diversity of antibiotic resistance genes in non-contaminated environments is much greater than in human and animal pathogens, and in environments contaminated with antibiotic from anthropogenic activities. This suggests the existence of some bottleneck effect for the mobilization of antibiotic resistance genes among different biomes. Bacteriophages have characteristics that make them suitable vectors between different biomes, and as well for transferring genes from biome to biome. Recent metagenomic studies and detection of bacterial genes by genomic techniques in the bacteriophage fraction of different microbiota provide indirect evidences that the mobilization of genes mediated by phages, including antibiotic resistance genes, is far more relevant than previously thought. Our hypothesis is that bacteriophages might be of critical importance for evading one of the bottlenecks, the lack of ecological connectivity that modulates the pass of antibiotic resistance genes from natural environments such as waters and soils, to animal and human microbiomes. This commentary concentrates on the potential importance of bacteriophages in transferring resistance genes from the environment to human and animal body microbiomes, but there is no doubt that transduction occurs also in body microbiomes.

  1. Foreign Body Infection Models to Study Host-Pathogen Response and Antimicrobial Tolerance of Bacterial Biofilm

    Directory of Open Access Journals (Sweden)

    Justyna Nowakowska

    2014-08-01

    Full Text Available The number of implanted medical devices is steadily increasing and has become an effective intervention improving life quality, but still carries the risk of infection. These infections are mainly caused by biofilm-forming staphylococci that are difficult to treat due to the decreased susceptibility to both antibiotics and host defense mechanisms. To understand the particular pathogenesis and treatment tolerance of implant-associated infection (IAI animal models that closely resemble human disease are needed. Applications of the tissue cage and catheter abscess foreign body infection models in the mouse will be discussed herein. Both models allow the investigation of biofilm and virulence of various bacterial species and a comprehensive insight into the host response at the same time. They have also been proven to serve as very suitable tools to study the anti-adhesive and anti-infective efficacy of different biomaterial coatings. The tissue cage model can additionally be used to determine pharmacokinetics, efficacy and cytotoxicity of antimicrobial compounds as the tissue cage fluid can be aspirated repeatedly without the need to sacrifice the animal. Moreover, with the advance in innovative imaging systems in rodents, these models may offer new diagnostic measures of infection. In summary, animal foreign body infection models are important tools in the development of new antimicrobials against IAI and can help to elucidate the complex interactions between bacteria, the host immune system, and prosthetic materials.

  2. Improving the In-Medium Similarity Renormalization Group via approximate inclusion of three-body effects

    Science.gov (United States)

    Morris, Titus; Bogner, Scott

    2016-09-01

    The In-Medium Similarity Renormalization Group (IM-SRG) has been applied successfully to the ground state of closed shell finite nuclei. Recent work has extended its ability to target excited states of these closed shell systems via equation of motion methods, and also complete spectra of the whole SD shell via effective shell model interactions. A recent alternative method for solving of the IM-SRG equations, based on the Magnus expansion, not only provides a computationally feasible route to producing observables, but also allows for approximate handling of induced three-body forces. Promising results for several systems, including finite nuclei, will be presented and discussed.

  3. Endotoxin-free purification for the isolation of Bovine Viral Diarrhoea Virus E2 protein from insoluble inclusion body aggregates

    Directory of Open Access Journals (Sweden)

    Mahony Timothy J

    2011-07-01

    Full Text Available Abstract Background Protein expression in Escherichia coli may result in the recombinant protein being expressed as insoluble inclusion bodies. In addition, proteins purified from E. coli contain endotoxins which need to be removed for in vivo applications. The structural protein, E2, from Bovine Viral Diarrhoea Virus (BVDV is a major immunogenic determinant, and is an ideal candidate as a subunit vaccine. The E2 protein contains 17 cysteine residues creating difficulties in E. coli expression. In this report we outline a procedure for successfully producing soluble and endotoxin-free BVDV E2 protein from inclusion bodies (IB. Results The expression of a truncated form of BVDV-E2 protein (E2-T1 in E. coli resulted in predominantly aggregated insoluble IB. Solubilisation of E2-T1 with high purity and stability from IB aggregates was achieved using a strong reducing buffer containing 100 mM Dithiothreitol. Refolding by dialysis into 50 mM Tris (pH 7.0 containing 0.2% Igepal CA630 resulted in a soluble but aggregated protein solution. The novel application of a two-phase extraction of inclusion body preparations with Triton X-114 reduced endotoxin in solubilised E2-T1 to levels suitable for in vivo use without affecting protein yields. Dynamic light scattering analyses showed 37.5% of the protein was monomeric, the remaining comprised of soluble aggregates. Mice immunised with E2-T1 developed a high titre antibody response by ELISA. Western hybridisation analysis showed E2-T1 was recognised by sera from immunised mice and also by several BVDV-E2 polyclonal and monoclonal antibodies. Conclusion We have developed a procedure using E. coli to produce soluble E2-T1 protein from IB, and due to their insoluble nature we utilised a novel approach using Triton X-114 to efficiently remove endotoxin. The resultant protein is immunogenic and detectable by BVDV-E2 specific antibodies indicating its usefulness for diagnostic applications and as a subunit

  4. A camelid nanobody against EGFR was easily obtained through refolding of inclusion body expressed in Escherichia coli.

    Science.gov (United States)

    Xu, Li; Song, Xiaoyu; Jia, Lingyun

    2017-11-01

    Using anti-EGFR (epidermal growth factor receptor) nanobody is a good choice for diagnoses and therapeutics for high EGFR expression diseases. In the present study, the percentage composition of anti-EGFR nanobody attained 25% of the total cell protein expressed in Escherichia coli BL21 (DE3). However, almost all nanobodies were expressed as inclusion bodies. To acquire active nanobodies, a series of dilution refolding procedures were optimized after inclusion bodies were dissolved into 6 M urea and purified with immobilized metal affinity chromatography. The results showed the refolding rate of the anti-EGFR nanobodies attained to 73%, and about 100 mg nanobodies were refolded from 1 L cells under the conditions that the initial nanobody concentration was 0.3 mg/mL, the dilution speed was 2.5 mL/Min, the dilution buffer was Tris-HCl at pH 8.0, the additives were 0.2 M Arg, 5 mM reduced glutathione (GSH), and 1 mM oxidized glutathione (GSSG). Then the activity of the refolded nanobodies was confirmed. The results showed that the refolded anti-EGFR nanobodies, in a dose-dependent manner, bounded to the tumor cell surface of A431 and MCF-7 and significantly inhibited the proliferation of A431 caused by the epidermal growth factor. Our study provides a facile method to rapidly, efficiently, and massively prepare anti-EGFR antibodies and promotes anti-EGFR-based recognition in cancer diagnoses and therapeutics. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

  5. A novel method to recover inclusion body protein from recombinant E. coli fed-batch processes based on phage ΦX174-derived lysis protein E.

    Science.gov (United States)

    Ehgartner, Daniela; Sagmeister, Patrick; Langemann, Timo; Meitz, Andrea; Lubitz, Werner; Herwig, Christoph

    2017-07-01

    Production of recombinant proteins as inclusion bodies is an important strategy in the production of technical enzymes and biopharmaceutical products. So far, protein from inclusion bodies has been recovered from the cell factory through mechanical or chemical disruption methods, requiring additional cost-intensive unit operations. We describe a novel method that is using a bacteriophage-derived lysis protein to directly recover inclusion body protein from Escherichia coli from high cell density fermentation process: The recombinant inclusion body product is expressed by using a mixed feed fed-batch process which allows expression tuning via adjusting the specific uptake rate of the inducing substrate. Then, bacteriophage ΦX174-derived lysis protein E is expressed to induce cell lysis. Inclusion bodies in empty cell envelopes are harvested via centrifugation of the fermentation broth. A subsequent solubilization step reveals the recombinant protein. The process was investigated by analyzing the impact of fermentation conditions on protein E-mediated cell lysis as well as cell lysis kinetics. Optimal cell lysis efficiencies of 99% were obtained with inclusion body titers of >2.0 g/l at specific growth rates higher 0.12 h -1 and inducer uptake rates below 0.125 g/(g × h). Protein E-mediated cell disruption showed a first-order kinetics with a kinetic constant of -0.8 ± 0.3 h -1 . This alternative inclusion body protein isolation technique was compared to the one via high-pressure homogenization. SDS gel analysis showed 10% less protein impurities when cells had been disrupted via high-pressure homogenization, than when empty cell envelopes including inclusion bodies were investigated. Within this contribution, an innovative technology, tuning recombinant protein production and substituting cost-intensive mechanical cell disruption, is presented. We anticipate that the presented method will simplify and reduce the production costs of inclusion body

  6. Immunohistochemical detection of a unique protein within cells of snakes having inclusion body disease, a world-wide disease seen in members of the families Boidae and Pythonidae.

    Directory of Open Access Journals (Sweden)

    Li-Wen Chang

    Full Text Available Inclusion body disease (IBD is a worldwide disease in captive boa constrictors (boa constrictor and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the IBD inclusion bodies, called IBD protein. A validated immuno-based ante-mortem diagnostic test is needed for screening snakes that are at risk of having IBD. In this study, despite difficulties in solubilizing semi-purified inclusion bodies, utilizing hybridoma technology a mouse anti-IBD protein monoclonal antibody (MAB was produced. The antigenic specificity of the antibody was confirmed and validated by western blots, enzyme-linked immunosorbent assay, immuno-transmission electron microscopy, and immunohistochemical staining. Paraffin embedded tissues of IBD positive and negative boa constrictors (n=94 collected from 1990 to 2011 were tested with immunohistochemical staining. In boa constrictors, the anti-IBDP MAB had a sensitivity of 83% and specificity of 100% in detecting IBD. The antibody also cross-reacted with IBD inclusion bodies in carpet pythons (Morelia spilota and a ball python (python regius. This validated antibody can serve as a tool for the development of ante-mortem immunodiagnostic tests for IBD.

  7. The disulfide-rich Metridia luciferase refolded from E. coli inclusion bodies reveals the properties of a native folded enzyme produced in insect cells.

    Science.gov (United States)

    Markova, Svetlana V; Larionova, Marina D; Gorbunova, Darya A; Vysotski, Eugene S

    2017-10-01

    The bioluminescence of a marine copepod Metridia longa is determined by a small secreted coelenterazine-dependent luciferase that uses coelenterazine as a substrate of enzymatic reaction to generate light (λ max =480nm). To date, four different isoforms of the luciferase differing in size, sequences, and properties have been cloned by functional screening. All of them contain ten conserved Cys residues that suggests up to five SS intramolecular bonds per luciferase molecule. Whereas the use of copepod luciferases as bioluminescent reporters in biomedical research in vivo is growing from year to year, their application for in vitro assays is still limited by the difficulty in obtaining significant amounts of luciferase. The most cost-effective host for producing recombinant proteins is Escherichia coli. However, prokaryotic and eukaryotic cells maintain the reductive environment in cytoplasm that hinders the disulfide bond formation and consequently the proper folding of luciferase. Here we report the expression of the MLuc7 isoform of M. longa luciferase in E. coli cells and the efficient procedure for refolding from inclusion bodies yielding a high-active monomeric protein. Furthermore, in a set of identical experiments we demonstrate that bioluminescent and structural features of MLuc7 produced in bacterial cells are identical to those of MLuc7 isoform produced from culture medium of insect cells. Although the yield of high-purity protein is only 6mg/L, the application of E. coli cells to produce the luciferase is simpler and more cost-effective than the use of insect cells. We expect that the suggested technology of Metridia luciferase production allows obtaining of sufficient amounts of protein both for the development of novel in vitro analytical assays with the use of MLuc7 as a label and for structural studies. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Data describing the inclusion relationships between two organs (PART-OF Tree) - BodyParts3D | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data List Contact us BodyParts...nclusion_relation_list.txt File URL: ftp://ftp.biosciencedbc.jp/archive/bodyparts3d/LATEST/partof_inclusion_...relation_list.txt File size: 90 KB Simple search URL http://togodb.biosciencedbc.jp/togodb/view/bodyparts3d_...ry of This Database Site Policy | Contact Us Data describing the inclusion relationships between two organs (PART-OF Tree) - BodyParts3D | LSDB Archive ...

  9. Data describing the inclusion relationships between two organs (IS-A Tree) - BodyParts3D | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data List Contact us BodyParts...relation_list.txt File URL: ftp://ftp.biosciencedbc.jp/archive/bodyparts3d/LATEST/isa_inclusion_relation_lis...t.txt File size: 203 KB Simple search URL http://togodb.biosciencedbc.jp/togodb/view/bodyparts3d_isa_inclusi...abase Site Policy | Contact Us Data describing the inclusion relationships between two organs (IS-A Tree) - BodyParts3D | LSDB Archive ...

  10. Cutaneous Bacterial Communities of a Poisonous Salamander: a Perspective from Life Stages, Body Parts and Environmental Conditions.

    Science.gov (United States)

    Sanchez, Eugenia; Bletz, Molly C; Duntsch, Laura; Bhuju, Sabin; Geffers, Robert; Jarek, Michael; Dohrmann, Anja B; Tebbe, Christoph C; Steinfartz, Sebastian; Vences, Miguel

    2017-02-01

    Amphibian skin provides a habitat for bacterial communities in its mucus. Understanding the structure and function of this "mucosome" in the European fire salamander (Salamandra salamandra) is critical in the context of novel emerging pathogenic diseases. We compare the cutaneous bacterial communities of this species using amplicon-based sequencing of the 16S rRNA V4 region. Across 290 samples, over 4000 OTUs were identified, four of them consistently present in all samples. Larvae and post-metamorphs exhibited distinct cutaneous microbial communities. In adults, the parotoid gland surface had a community structure different from the head, dorsum, flanks and ventral side. Larvae from streams had higher phylogenetic diversity than those found in ponds. Their bacterial community structure also differed; species of Burkholderiaceae, Comamonadaceae, Methylophilaceae and Sphingomonadaceae were more abundant in pond larvae, possibly related to differences in factors like desiccation and decomposition rate in this environment. The observed differences in the cutaneous bacterial community among stages, body parts and habitats of fire salamanders suggest that both host and external factors shape these microbiota. We hypothesize that the variation in cutaneous bacterial communities might contribute to variation in pathogen susceptibility among individual salamanders.

  11. Characterization of the 4,6-α-glucanotransferase GTFB enzyme of Lactobacillus reuteri 121 isolated from inclusion bodies.

    Science.gov (United States)

    Bai, Yuxiang; van der Kaaij, Rachel Maria; Woortman, Albert Jan Jacob; Jin, Zhengyu; Dijkhuizen, Lubbert

    2015-06-09

    The GTFB enzyme of the probiotic bacterium Lactobacillus reuteri 121 is a 4,6-α-glucanotransferase of glycoside hydrolase family 70 (GH70; http://www.cazy.org ). Contrary to the glucansucrases in GH70, GTFB is unable to use sucrose as substrate, but instead converts malto-oligosaccharides and starch into isomalto-/malto- polymers that may find application as prebiotics and dietary fibers. The GTFB enzyme expresses well in Escherichia coli BL21 Star (DE3), but mostly accumulates in inclusion bodies (IBs) which generally contain wrongly folded protein and inactive enzyme. Denaturation followed by refolding, as well as ncIB preparation were used for isolation of active GTFB protein from inclusion bodies. Soluble, refolded and ncIB GTFB were compared using activity assays, secondary structure analysis by FT-IR, and product analyses by NMR, HPAEC and SEC. Expression of GTFB in E. coli yielded > 100 mg/l relatively pure and active but mostly insoluble GTFB protein in IBs, regardless of the expression conditions used. Following denaturing, refolding of GTFB protein was most efficient in double distilled H2O. Also, GTFB ncIBs were active, with approx. 10 % of hydrolysis activity compared to the soluble protein. When expressed as units of activity obtained per liter E. coli culture, the total amount of ncIB GTFB expressed possessed around 180 % hydrolysis activity and 100 % transferase activity compared to the amount of soluble GTFB enzyme obtained from one liter culture. The product profiles obtained for the three GTFB enzyme preparations were similar when analyzed by HPAEC and NMR. SEC investigation also showed that these 3 enzyme preparations yielded products with similar size distributions. FT-IR analysis revealed extended β-sheet formation in ncIB GTFB providing an explanation at the molecular level for reduced GTFB activity in ncIBs. The thermostability of ncIB GTFB was relatively high compared to the soluble and refolded GTFB. In view of their relatively high yield

  12. Upon Infection the Cellular WD Repeat-containing Protein 5 (WDR5) Localizes to Cytoplasmic Inclusion Bodies and Enhances Measles Virus Replication.

    Science.gov (United States)

    Ma, Dzwokai; George, Cyril X; Nomburg, Jason; Pfaller, Christian K; Cattaneo, Roberto; Samuel, Charles E

    2017-12-13

    Replication of negative-strand RNA viruses occurs in association with discrete cytoplasmic foci called inclusion bodies. Whereas inclusion bodies represent a prominent subcellular structure induced by viral infection, our knowledge of the cellular protein components involved in inclusion body formation and function is limited. Using measles virus-infected HeLa cells, we found that the WD repeat-containing protein 5 (WDR5), a subunit of histone H3 lysine 4 methyltransferases, was selectively recruited to virus-induced inclusion bodies. Furthermore, WDR5 was found in complexes containing viral proteins associated with RNA replication. WDR5 was not detected with mitochondria, stress granules, or other known secretory or endocytic compartments of infected cells. WDR5 deficiency decreased both viral protein production and infectious virus yields. Interferon production was modestly increased in WDR5 deficient cells. Thus, our study identifies WDR5 as a novel viral inclusion body-associated cellular protein and suggests a role for WDR5 in promoting viral replication. IMPORTANCE Measles virus is a human pathogen that remains a global concern with more than 100,000 measles-related deaths annually despite the availability of an effective vaccine. As measles continues to cause significant morbidity and mortality, understanding the virus-host interactions at the molecular level that affect virus replication efficiency is important for development and optimization of treatment procedures. Measles virus is an RNA virus that encodes six genes and replicates in the cytoplasm of infected cells in discrete cytoplasmic replication bodies, though little is known of the biochemical nature of these structures. Here we show that the cellular protein WDR5 is enriched in the cytoplasmic viral replication factories and enhances virus growth. WDR5-containing protein complex includes viral proteins responsible for viral RNA replication. Thus, we have identified WDR5 as a host factor that

  13. Sporadic inclusion-body myositis: A degenerative muscle disease associated with aging, impaired muscle protein homeostasis and abnormal mitophagy.

    Science.gov (United States)

    Askanas, Valerie; Engel, W King; Nogalska, Anna

    2015-04-01

    Sporadic inclusion-body myositis (s-IBM) is the most common degenerative muscle disease in which aging appears to be a key risk factor. In this review we focus on several cellular molecular mechanisms responsible for multiprotein aggregation and accumulations within s-IBM muscle fibers, and their possible consequences. Those include mechanisms leading to: a) accumulation in the form of aggregates within the muscle fibers, of several proteins, including amyloid-β42 and its oligomers, and phosphorylated tau in the form of paired helical filaments, and we consider their putative detrimental influence; and b) protein misfolding and aggregation, including evidence of abnormal myoproteostasis, such as increased protein transcription, inadequate protein disposal, and abnormal posttranslational modifications of proteins. Pathogenic importance of our recently demonstrated abnormal mitophagy is also discussed. The intriguing phenotypic similarities between s-IBM muscle fibers and the brains of Alzheimer and Parkinson's disease patients, the two most common neurodegenerative diseases associated with aging, are also discussed. This article is part of a Special Issue entitled: Neuromuscular Diseases: Pathology and Molecular Pathogenesis. Copyright © 2014. Published by Elsevier B.V.

  14. Mutant LRRK2 toxicity in neurons depends on LRRK2 levels and synuclein but not kinase activity or inclusion bodies.

    Science.gov (United States)

    Skibinski, Gaia; Nakamura, Ken; Cookson, Mark R; Finkbeiner, Steven

    2014-01-08

    By combining experimental neuron models and mathematical tools, we developed a "systems" approach to deconvolve cellular mechanisms of neurodegeneration underlying the most common known cause of Parkinson's disease (PD), mutations in leucine-rich repeat kinase 2 (LRRK2). Neurons ectopically expressing mutant LRRK2 formed inclusion bodies (IBs), retracted neurites, accumulated synuclein, and died prematurely, recapitulating key features of PD. Degeneration was predicted from the levels of diffuse mutant LRRK2 that each neuron contained, but IB formation was neither necessary nor sufficient for death. Genetic or pharmacological blockade of its kinase activity destabilized LRRK2 and lowered its levels enough to account for the moderate reduction in LRRK2 toxicity that ensued. By contrast, targeting synuclein, including neurons made from PD patient-derived induced pluripotent cells, dramatically reduced LRRK2-dependent neurodegeneration and LRRK2 levels. These findings suggest that LRRK2 levels are more important than kinase activity per se in predicting toxicity and implicate synuclein as a major mediator of LRRK2-induced neurodegeneration.

  15. Cloning and expression of phosphoenolpyruvate carboxykinase from a cestode parasite and its solubilization from inclusion bodies using l-arginine.

    Science.gov (United States)

    Dutta, Asim K; Ramnath; Dkhar, Barilin; Tandon, Veena; Das, Bidyadhar

    2016-09-01

    Phosphoenolpyruvate carboxykinase is an essential regulatory enzyme of glycolysis in the cestode parasite, Raillietina echinobothrida, and is considered a potential target for anthelmintic action because of its differential activity from that of its avian host. However, due to the unavailability of its structure, the mechanism of regulation of PEPCK from R. echinobothrida (rePEPCK) and its interaction with possible modulators remain unclear. Hence, in this study, the rePEPCK gene was cloned into pGEX-4T-3 and overexpressed for its characterization. On being induced by IPTG, the recombinant rePEPCK was expressed as inclusion bodies (IBs); hence, various agents, like different inducer concentrations, temperature, time, host cell types, culture media, pH, and additives, were used to bring the protein to soluble form. Finally, a significant amount (∼46%) of rePEPCK was solubilized from IBs by adding 2M l-arginine. Near-UV circular dichroism spectra analysis indicated that l-arginine (2M) had no effect on the conformation of the protein. In this study, we have reported a yield of ∼73mg of purified rePEPCK per 1L of culture. The purified rePEPCK retained its biological activity, and Km of the enzyme for its substrate was determined and discussed. The availability of recombinant rePEPCK may help in biochemical- and biophysical-studies to explore its molecular mechanisms and regulations. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. Reassessment of inclusion body-based production as a versatile opportunity for difficult-to-express recombinant proteins.

    Science.gov (United States)

    Hoffmann, Daniel; Ebrahimi, Mehrdad; Gerlach, Doreen; Salzig, Denise; Czermak, Peter

    2017-11-10

    The production of recombinant proteins in the microbial host Escherichia coli often results in the formation of cytoplasmic protein inclusion bodies (IBs). Proteins forming IBs are often branded as difficult-to-express, neglecting that IBs can be an opportunity for their production. IBs are resistant to proteolytic degradation and contain up to 90% pure recombinant protein, which does not interfere with the host metabolism. This is especially advantageous for host-toxic proteins like antimicrobial peptides (AMPs). IBs can be easily isolated by cell disruption followed by filtration and/or centrifugation, but conventional techniques for the recovery of soluble proteins from IBs are laborious. New approaches therefore simplify protein recovery by optimizing the production process conditions, and often include mild resolubilization methods that either increase the yield after refolding or avoid the necessity of refolding all together. For the AMP production, the IB-based approach is ideal, because these peptides often have simple structures and are easy to refold. The intentional IB production of almost every protein can be achieved by fusing recombinant proteins to pull-down tags. This review discusses the techniques available for IB-based protein production before considering technical approaches for the isolation of IBs from E. coli lysates followed by efficient protein resolubilization which ideally omits further refolding. The techniques are evaluated in terms of their suitability for the process-scale production and downstream processing of recombinant proteins and are discussed for AMP production as an example.

  17. Psychological Impact of Predictive Genetic Testing in VCP Inclusion Body Myopathy, Paget Disease of Bone and Frontotemporal Dementia.

    Science.gov (United States)

    Surampalli, Abhilasha; Khare, Manaswitha; Kubrussi, Georgette; Wencel, Marie; Tanaja, Jasmin; Donkervoort, Sandra; Osann, Kathryn; Simon, Mariella; Wallace, Douglas; Smith, Charles; M McInerney-Leo, Aideen; Kimonis, Virginia

    2015-10-01

    Inclusion Body Myopathy associated with Paget's disease of bone and Fronto-temporal Dementia, also known as multisystem proteinopathy is an autosomal dominant, late onset neurodegenerative disorder caused by mutations in Valosin containing protein (VCP) gene. This study aimed to assess uptake and decision making for predictive genetic testing and the impact on psychological well-being. Individuals who had participated in the gene discovery study with a 50 % a priori risk of inheriting VCP disease were sent a letter of invitation offering genetic counseling and testing and were also invited to participate in this psychosocial study. A total of 102 individuals received an invitation and 33 individuals participated in genetic counseling and testing (32.3 %) with 29 completing baseline questionnaires. Twenty completed the follow-up post-test Hospital Anxiety and Depression Scale questionnaire including 13 of the 18 who had tested positive. Mean risk perception at baseline was 50.1 %. Reasons for testing included planning for the future, relieving uncertainty, informing children and satisfying curiosity. At baseline, one quarter of the participants had high levels of anxiety. However, scores were normal one year following testing. In this small cohort, one third of individuals at 50 % risk chose pre-symptomatic testing. Although one quarter of those choosing testing had high anxiety at baseline, this was not evident at follow-up.

  18. Evasion of Antiviral Immunity through Sequestering of TBK1/IKKε/IRF3 into Viral Inclusion Bodies

    Science.gov (United States)

    Wu, Xiaodong; Qi, Xian; Qu, Bingqian; Zhang, Zerui; Liang, Mifang; Li, Chuan; Cardona, Carol J.; Li, Dexin

    2014-01-01

    Cells are equipped with pattern recognition receptors (PRRs) such as the Toll-like and RIG-I-like receptors that mount innate defenses against viruses. However, viruses have evolved multiple strategies to evade or thwart host antiviral responses. Viral inclusion bodies (IBs), which are accumulated aggregates of viral proteins, are commonly formed during the replication of some viruses in infected cells, but their role in viral immune evasion has rarely been explored. Severe fever with thrombocytopenia syndrome (SFTS) is an emerging febrile illness caused by a novel phlebovirus in the Bunyaviridae. The SFTS viral nonstructural protein NSs can suppress host beta interferon (IFN-β) responses. NSs can form IBs in infected and transfected cells. Through interaction with tank-binding kinase 1 (TBK1), viral NSs was able to sequester the IKK complex, including IKKε and IRF3, into IBs, although NSs did not interact with IKKε or IRF3 directly. When cells were infected with influenza A virus, IRF3 was phosphorylated and active phosphorylated IRF3 (p-IRF3) was translocated into the nucleus. In the presence of NSs, IRF3 could still be phosphorylated, but p-IRF3 was trapped in cytoplasmic IBs, resulting in reduced IFN-β induction and enhanced viral replication. Sequestration of the IKK complex and active IRF3 into viral IBs through the interaction of NSs and TBK1 is a novel mechanism for viral evasion of innate immunity. PMID:24335286

  19. High pH solubilization and chromatography-based renaturation and purification of recombinant human granulocyte colony-stimulating factor from inclusion bodies.

    Science.gov (United States)

    Li, Ming; Fan, Hua; Liu, Jiahua; Wang, Minhong; Wang, Lili; Wang, Chaozhan

    2012-03-01

    Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is a very efficient therapeutic protein drug which has been widely used in human clinics to treat cancer patients suffering from chemotherapy-induced neutropenia. In this study, rhG-CSF was solubilized from inclusion bodies by using a high-pH solution containing low concentration of urea. It was found that solubilization of the rhG-CSF inclusion bodies greatly depended on the buffer pH employed; alkalic pH significantly favored the solubilization. In addition, when small amount of urea was added to the solution at high pH, the solubilization was further enhanced. After solubilization, the rhG-CSF was renatured with simultaneous purification by using weak anion exchange, strong anion exchange, and hydrophobic interaction chromatography, separately. The results indicated that the rhG-CSF solubilized by the high-pH solution containing low concentration of urea had much higher mass recovery than the one solubilized by 8 M urea when using anyone of the three refolding methods employed in this work. In the case of weak anion exchange chromatography, the high pH solubilized rhG-CSF could get a mass recovery of 73%. The strategy of combining solubilization of inclusion bodies at high pH with refolding of protein using liquid chromatography may become a routine method for protein production from inclusion bodies.

  20. Estudo comparativo das inclusões do alastrim e da vacina no macaco (Macacus rhesus A comparison of the inclusion bodies of alastrim and vaccinia in the monkey (Macacus rhesus

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    C. Magarinos Torres

    1934-02-01

    Full Text Available Vesiculas e pustulas contendo numerosas inclusões citoplasmicas nas celulas epidermicas, foram regularmente produzidas no macaco (Macacus rhesus, quer com o virus do alastrim, quer com o da vacina, após inoculação endovenosa e sem previa escarificação. O virus do alastrim parece menos virulento para essa especie de macaco que o da vacina. Ao passo que 12 macacos rhesus injetados por via endovenosa com sete amostras diferentes de virus do alastrim, após apresentarem com regularidade um infecção experimental, sobreviveram e se conservaram em boa saúde, a injecção endovenosa do virus da vacina recentemente preparado (polpa bruta produziu a morte em 2, dentre 4 animais experimentados. 2. - Foram notadas diferenças pequenas, mas nitidas, na morfologia das inclusões do alastrim e da vacina, em material fixado no liquido de Helly, incluido em parafina e corado pela hematoxilina-eosina. Dizem elas respeito ao numero de inclusões encontradas em cada celula epidermica e às suas reações de coloração. 3. - As inclusões do alastrim, quando apresentam grandes dimensões, conservam-se unicas ou solitarias no citoplasma das celulas epidermicas do macaco rhesus, e coram-se em tonalidade que varia do azul escuro ao cinzento-azulado. Comtudo, em celulas que sofreram necrose, ou naquelas contendo 2 a 4 inclusões de pequenas dimensões, por vezes elas se mostram coradas em roseo. 4. - As inclusões da vacina, quando em faze adeantada de desenvolvimento, são multiplas nas celulas epidermicas do macaco rhesus e mostram, regularmente, uma policromatofilia caracteristica.1. - Vesicles and pustules containing numerous cytoplasmic inclusion bodies within the epidermal cells were regularly produced in monkeys (Macacus rhesus by intravenous inoculation either of alastrim virus or of recently prepared vaccine emulsion, no previous scarifications being required. Alastrim virus seems less virulent for this species of monkey than the virus of vaccinia is

  1. Prevalence of Howell-Jolly body-like inclusions in HIV patients and their correlation with CD4 counts and HIV RNA viral load.

    Science.gov (United States)

    Chang, Brian; Huang, Richard Sheng Poe; Dasgupta, Amitava; Nguyen, Nghia; Wahed, Amer

    2015-01-01

    Previous reports have described the rare occurrence of detached nuclear fragments resembling Howell-Jolly bodies within neutrophils from HIV patients, organ-transplant recipients, and patients on immunosuppressive drugs. To date, their potential clinical significance is unknown, and pathologists tend to disregard their presence. Our study sought to find a correlation between these inclusions and the overall disease state, specifically within the HIV patient population. Eighty-three peripheral smears, all from different patients, were examined for the presence of inclusions and compared with recent CD4 counts and HIV RNA viral loads. Six cases contained inclusions, yielding a prevalence of 7.2%. These six patients had a mean CD4 count of 546±305 cells/μL compared to 247±242 cells/μL in those lacking inclusions (pHowell-Jolly body-like inclusions may be viewed as a potential biomarker indicative of a low risk for disease progression and/or good response to therapy based upon higher CD4 counts and relatively favorable viral loads. © 2015 by the Association of Clinical Scientists, Inc.

  2. High hydrostatic pressure enables almost 100% refolding of recombinant human ciliary neurotrophic factor from inclusion bodies at high concentration.

    Science.gov (United States)

    Wang, Qi; Liu, Yongdong; Zhang, Chun; Guo, Fangxia; Feng, Cui; Li, Xiunan; Shi, Hong; Su, Zhiguo

    2017-05-01

    Protein refolding from inclusion bodies (IBs) often encounters a problem of low recovery at high protein concentration. In this study, we demonstrated that high hydrostatic pressure (HHP) could simultaneously achieve high refolding concentration and high refolding yield for IBs of recombinant human ciliary neurotrophic factor (rhCNTF), a potential therapeutic for neurodegenerative diseases. The use of dilution refolding obtained 18% recovery at 3 mg/mL, even in the presence of 4 M urea. In contrast, HHP refolding could efficiently increase the recovery up to almost 100% even at 4 mg/mL. It was found that in the dilution, hydrophobic aggregates were the off-path products and their amount increased with the protein concentration. However, HHP could effectively minimize the formation of hydrophobic aggregates, leading to almost complete conversion of the rhCNTF IBs to the correct configuration. The stable operation range of concentration is 0.5-4.0 mg/mL, in which the refolding yield was almost 100%. Compared with the literatures where HHP failed to increase the refolding yield beyond 90%, the reason could be attributed to the structural difference that rhCNTF has no disulfide bond and is a monomeric protein. After purification by one-step of anionic chromatography, the purity of rhCNTF reached 95% with total process recovery of 54.1%. The purified rhCNTF showed similar structure and in vitro bioactivity to the native species. The whole process featured integration of solubilization/refolding, a high refolding yield of 100%, a high concentration of 4 mg/mL, and a simple chromatography to ensure a high productivity. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Disease specificity of autoantibodies to cytosolic 5'-nucleotidase 1A in sporadic inclusion body myositis versus known autoimmune diseases.

    Science.gov (United States)

    Herbert, Megan K; Stammen-Vogelzangs, Judith; Verbeek, Marcel M; Rietveld, Anke; Lundberg, Ingrid E; Chinoy, Hector; Lamb, Janine A; Cooper, Robert G; Roberts, Mark; Badrising, Umesh A; De Bleecker, Jan L; Machado, Pedro M; Hanna, Michael G; Plestilova, Lenka; Vencovsky, Jiri; van Engelen, Baziel G; Pruijn, Ger J M

    2016-04-01

    The diagnosis of inclusion body myositis (IBM) can be challenging as it can be difficult to clinically distinguish from other forms of myositis, particularly polymyositis (PM). Recent studies have shown frequent presence of autoantibodies directed against cytosolic 5'-nucleotidase 1A (cN-1A) in patients with IBM. We therefore, examined the autoantigenicity and disease specificity of major epitopes of cN-1A in patients with sporadic IBM compared with healthy and disease controls. Serum samples obtained from patients with IBM (n=238), PM and dermatomyositis (DM) (n=185), other autoimmune diseases (n=246), other neuromuscular diseases (n=93) and healthy controls (n=35) were analysed for the presence of autoantibodies using immunodominant cN-1A peptide ELISAs. Autoantibodies directed against major epitopes of cN-1A were frequent in patients with IBM (37%) but not in PM, DM or non-autoimmune neuromuscular diseases (autoimmune diseases, particularly Sjögren's syndrome (SjS; 36%) and systemic lupus erythematosus (SLE; 20%). In summary, we found frequent anti-cN-1A autoantibodies in sera from patients with IBM. Heterogeneity in reactivity with the three immunodominant epitopes indicates that serological assays should not be limited to a distinct epitope region. The similar reactivities observed for SjS and SLE demonstrate the need to further investigate whether distinct IBM-specific epitopes exist. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  4. Kinetics of viral load and erythrocytic inclusion body formation in pacific herring artificially infected with erythrocytic necrosis virus

    Science.gov (United States)

    Glenn, Jolene A.; Emmenegger, Eveline J.; Grady, Courtney A.; Roon, Sean R.; Gregg, Jacob L.; Conway, Carla M.; Winton, James R.; Hershberger, Paul K.

    2012-01-01

    Viral erythrocytic necrosis (VEN) is a condition that affects marine and anadromous fish species, including herrings and salmonids, in the Atlantic and Pacific oceans. Infection is frequently associated with severe anemia and causes episodic mortality among wild and hatchery fish when accompanied by additional stressors; VEN can be presumptively diagnosed by (1) light microscopic identification of a single characteristic—a round, magenta-colored, 0.8-μm-diameter inclusion body (IB) within the cytoplasm of erythrocytes and their precursors on Giemsa-stained blood films; or (2) observation (via transmission electron microscopy [TEM]) of the causative iridovirus, erythrocytic necrosis virus (ENV), within erythrocytes or their precursors. To better understand the kinetics of VEN, specific-pathogen-free Pacific herring Clupea pallasii were infected with ENV by intraperitoneal injection. At 1, 4, 7, 10, 14, 21, and 28 d postexposure, samples of blood, spleen, and kidney were collected and assessed (1) via light microscopy for the number of intracytoplasmic IBs in blood smears and (2) via TEM for the number of virions within erythrocytes. The mean prevalence of intracytoplasmic IBs in the blood cells increased from 0% at 0–4 d postexposure to 94% at 28 d postexposure. Viral load within circulating red blood cells peaked at 7 d postexposure, fell slightly, and then reached a plateau. However, blood cells observed within the kidney and spleen tissues demonstrated high levels of ENV between 14 and 28 d postexposure. The results indicate that the viral load within erythrocytes does not correlate well with IB prevalence and that the virus can persist in infected fish for more than 28 d.

  5. Corpos de inclusão citoplasmática: estudo em diversas doenças e revisão da literatura Inclusion cytoplasmic bodies: a study in several diseases and a literature review

    Directory of Open Access Journals (Sweden)

    Rosana Herminia Scola

    1996-06-01

    Full Text Available Estudamos 16 casos entre 1400 biópsias musculares que apresentavam vacúolos marginados, cujo aspecto histológico sugeria corpos de inclusão citoplasmáticos. Procuramos correlacionar os dados clínicos, laboratoriais e histopatológicos, a fim de determinar a especificidade dos corpos de inclusão citoplasmáticos para determinadas doenças. A creatinaquinase mostrou-se elevada em 10 casos. A eletromiografia foi anormal em todos os casos. A histoquímica muscular em 5 casos revelou uma miopatia, em 7 padrão misto, em dois desinervação e em 2 casos miopatia inflamatória. A microscopia eletrônica demonstrou a presença de filamentos em 8 casos (nucleares, dispersos no citoplasma ou na região subsarcolemal. Os pacientes foram classificados conforme a história clínica, hereditariedade, dados laboratoriais, eletrofisiológicos, histoquímicos e microscopia eletrônica Encontramos miosite com corpos de inclusão citoplasmática (4 casos, atrofia muscular espinhal juvenil (6 casos, miopatias distais (3 casos, distrofia de cinturas pélvica e escapular (2 casos e polineuropatia periférica (1 caso. Apresentamos revisão sobre a patogenia, formação e possível etiologia dos vacúolos marginados e sua relação com as diversas entidades em que foram detectados, sugerindo que não são específicos para uma única doença.Among 1400 muscle biopsies, we studied 16 cases with rimmed vacuoles, whose histology suggests cytoplasm inclusion bodies. We tried to correlate the clinical, laboratory and histopatological data in order to verify the specificity of cytoplasm inclusion bodies to certain diseases. The creatinekinase was increased in 10 cases. In all cases electromyography was abnormal. Muscle histochemistry revealed myopathy in 5 cases, mixed pattern in 7, denervation in 2 and in 2 cases, inflammatory myopathy. Electron microscopy showed the presence of filaments in 8 cases (nuclear, disseminated in cytoplasm or in the subsarcolemmal region

  6. Pasteurization Procedures for Donor Human Milk Affect Body Growth, Intestinal Structure, and Resistance against Bacterial Infections in Preterm Pigs.

    Science.gov (United States)

    Li, Yanqi; Nguyen, Duc Ninh; de Waard, Marita; Christensen, Lars; Zhou, Ping; Jiang, Pingping; Sun, Jing; Bojesen, Anders Miki; Lauridsen, Charlotte; Lykkesfeldt, Jens; Dalsgaard, Trine Kastrup; Bering, Stine Brandt; Sangild, Per Torp

    2017-06-01

    Background: Holder pasteurization (HP) destroys multiple bioactive factors in donor human milk (DM), and UV-C irradiation (UVC) is potentially a gentler method for pasteurizing DM for preterm infants. Objective: We investigated whether UVC-treated DM improves gut maturation and resistance toward bacterial infections relative to HP-treated DM. Methods: Bacteria, selected bioactive components, and markers of antioxidant capacity were measured in unpasteurized donor milk (UP), HP-treated milk, and UVC-treated milk (all from the same DM pool). Fifty-seven cesarean-delivered preterm pigs (91% gestation; ratio of males to females, 30:27) received decreasing volumes of parental nutrition (average 69 mL · kg -1 · d -1 ) and increasing volumes of the 3 DM diets ( n = 19 each, average 89 mL · kg -1 · d -1 ) for 8-9 d. Body growth, gut structure and function, and systemic bacterial infection were evaluated. Results: A high bacterial load in the UP (6×10 5 colony forming units/mL) was eliminated similarly by HP and UVC treatments. Relative to HP-treated milk, both UVC-treated milk and UP showed greater activities of lipase and alkaline phosphatase and concentrations of lactoferrin, secretory immunoglobulin A, xanthine dehydrogenase, and some antioxidant markers (all P bacterial cultures in the bone marrow (28%) than pigs fed HP-treated milk (68%) ( P resistance against bacterial infections as shown in preterm pigs as a model for DM-fed preterm infants. © 2017 American Society for Nutrition.

  7. MB109 as bioactive human bone morphogenetic protein-9 refolded and purified from E. coli inclusion bodies

    Science.gov (United States)

    2014-01-01

    Background The development of chemical refolding of transforming growth factor-beta (TGF-β) superfamily ligands has been instrumental to produce the recombinant proteins for biochemical studies and exploring the potential of protein therapeutics. The osteogenic human bone morphogenetic protein-2 (hBMP-2) and its Drosophila DPP homolog were the early successful cases of refolding into functional form. Despite the similarity in their three dimensional structure and amino acid sequences, several other TGF-β superfamily ligands could not be refolded readily by the same methods. Results Here, we report a comprehensive study on the variables of a rapid-dilution refolding method, including the concentrations of protein, salt, detergent and redox agents, pH, refolding duration and the presence of aggregation suppressors and host-cell contaminants, in order to identify the optimal condition to refold human BMP-9 (hBMP-9). To produce a recombinant form of hBMP-9 in E. coli cells, a synthetic codon-optimized gene was designed to encode the mature domain of hBMP-9 (Ser320 – Arg429) directly behind the first methionine, which we herein referred to as MB109. An effective purification scheme was also developed to purify the refolded MB109 to homogeneity with a final yield of 7.8 mg from 100 mg of chromatography-purified inclusion bodies as a starting material. The chemically refolded MB109 binds to ALK1, ActRIIb and BMPRII receptors with relatively high affinity as compared to other Type I and Type II receptors based on surface plasmon resonance analysis. Smad1-dependent luciferase assay in C2C12 cells shows that the MB109 has an EC50 of 0.61 ng/mL (25 pM), which is nearly the same as hBMP-9. Conclusion MB109 is prone to be refolded as non-functional dimer and higher order multimers in most of the conditions tested, but bioactive MB109 dimer can be refolded with high efficiency in a narrow window, which is strongly dependent on the pH, refolding duration, the presence of

  8. Inclusion of Regional Body Composition Parameters Improves Bone Mineral Density Cross-Calibration Between GE Lunar Prodigy and iDXA Densitometers.

    Science.gov (United States)

    Saarelainen, J; Hakulinen, M; Rikkonen, T; Kröger, H; Koivumaa-Honkanen, H; Honkanen, R; Jurvelin, J S

    Since 1989, the Kuopio Osteoporosis Risk Factor and Prevention (OSTPRE, n = 14220) Study has followed long-term changes of bone mineral density (BMD) and body composition in women with GE Lunar devices. During the course of OSTPRE, the dual-energy X-ray absorptiometry device had to be replaced by a newer model. Then, it was essential to determine whether systematic measurement differences in BMD and body composition will occur. As a part of the OSTPRE study, BMD was measured in 54 women, whereas body composition was determined in 55 women, aged 27-71, by using both the GE Healthcare Lunar Prodigy and iDXA narrow-angle fan beam densitometers during the same visit. The total body fat mass (FM) and lean body mass (LBM) results of these scanners showed a high linear correlation (r = 0.981-0.994, p Prodigy. Inclusion of local soft tissue measurements (total body LBM, legs/android FM) improved the agreement of total body, total hip, and lumbar spine BMD values between the devices but not femoral neck BMD agreement. Equations, based on linear regression analyses, were derived to minimize differences between the instruments. Then, the differences in BMD and body composition measurements were negligible between Prodigy and iDXA. Using correction equations enables an objective comparison of longitudinal BMD and body composition measurements. Copyright © 2016 The International Society for Clinical Densitometry. Published by Elsevier Inc. All rights reserved.

  9. The fibrous form of intracellular inclusion bodies in recombinant variant fibrinogen-producing cells is specific to the hepatic fibrinogen storage disease-inducible variant fibrinogen.

    Science.gov (United States)

    Arai, Shinpei; Ogiwara, Naoko; Mukai, Saki; Takezawa, Yuka; Sugano, Mitsutoshi; Honda, Takayuki; Okumura, Nobuo

    2017-06-01

    Fibrinogen storage disease (FSD) is a rare disorder that is characterized by the accumulation of fibrinogen in hepatocytes and induces liver injury. Six mutations in the γC domain (γG284R, γT314P, γD316N, the deletion of γG346-Q350, γG366S, and γR375W) have been identified for FSD. Our group previously established γ375W fibrinogen-producing Chinese hamster ovary (CHO) cells and observed aberrant large granular and fibrous forms of intracellular inclusion bodies. The aim of this study was to investigate whether fibrous intracellular inclusion bodies are specific to FSD-inducible variant fibrinogen. Thirteen expression vectors encoding the variant γ-chain were stably or transiently transfected into CHO cells expressing normal fibrinogen Aα- and Bβ-chains or HuH-7 cells, which were then immunofluorescently stained. Six CHO and HuH-7 cell lines that transiently produced FSD-inducible variant fibrinogen presented the fibrous (3.2-22.7 and 2.1-24.5%, respectively) and large granular (5.4-25.5 and 7.7-23.9%) forms of intracellular inclusion bodies. Seven CHO and HuH-7 cell lines that transiently produced FSD-non-inducible variant fibrinogen only exhibit the large granular form. These results demonstrate that transiently transfected variant fibrinogen-producing CHO cells and inclusion bodies of the fibrous form may be useful in non-invasive screening for FSD risk factors for FSD before its onset.

  10. Body lice of homeless people reveal the presence of several emerging bacterial pathogens in northern Algeria.

    Science.gov (United States)

    Louni, Meriem; Mana, Nassima; Bitam, Idir; Dahmani, Mustapha; Parola, Philippe; Fenollar, Florence; Raoult, Didier; Mediannikov, Oleg

    2018-04-17

    Human lice, Pediculus humanus, are obligate blood-sucking parasites. Body lice, Pediculus h. humanus, occur in two divergent mitochondrial clades (A and D) each exhibiting a particular geographic distribution. Currently, the body louse is recognized as the only vector for louse-borne diseases. In this study, we aimed to study the genetic diversity of body lice collected from homeless populations in three localities of northern Algeria, and to investigate louse-borne pathogens in these lice. In this study, 524 body lice specimens were collected from 44 homeless people in three localities: Algiers, Tizi Ouzou and Boumerdès located in northern Algeria. Duplex clade specific real-time PCRs (qPCR) and Cytochrome b (cytb) mitochondrial DNA (mtDNA) analysis were performed in order to identify the mitochondrial clade. Screening of louse-borne pathogens bacteria was based on targeting specific genes for each pathogen using qPCR supplemented by sequencing. All body lice belong to clade A. Through amplification and sequencing of the cytb gene we confirmed the presence of three haplotypes: A5, A9 and A63, which is novel. The molecular investigation of the 524 body lice samples revealed the presence of four human pathogens: Bartonella quintana (13.35%), Coxiella burnetii (10.52%), Anaplasma phagocytophilum (0.76%) and Acinetobacter species (A. baumannii, A. johnsonii, A. berezeniae, A. nosocomialis and A. variabilis, in total 46.94%). To the best of our knowledge, our study is the first to show the genetic diversity and presence of several emerging pathogenic bacteria in homeless' body lice from Algeria. We also report for the first time, the presence of several species of Acinetobacter in human body lice. Our results highlight the fact that body lice may be suspected as being a much broader vector of several pathogenic agents than previously thought. Nevertheless, other studies are needed to encourage epidemiological investigations and surveys of louse-associated infections.

  11. VCP associated inclusion body myopathy and paget disease of bone knock-in mouse model exhibits tissue pathology typical of human disease.

    Directory of Open Access Journals (Sweden)

    Mallikarjun Badadani

    2010-10-01

    Full Text Available Dominant mutations in the valosin containing protein (VCP gene cause inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia (IBMPFD. We have generated a knock-in mouse model with the common R155H mutation. Mice demonstrate progressive muscle weakness starting approximately at the age of 6 months. Histology of mutant muscle showed progressive vacuolization of myofibrils and centrally located nuclei, and immunostaining shows progressive cytoplasmic accumulation of TDP-43 and ubiquitin-positive inclusion bodies in quadriceps myofibrils and brain. Increased LC3-II staining of muscle sections representing increased number of autophagosomes suggested impaired autophagy. Increased apoptosis was demonstrated by elevated caspase-3 activity and increased TUNEL-positive nuclei. X-ray microtomography (uCT images show radiolucency of distal femurs and proximal tibiae in knock-in mice and uCT morphometrics shows decreased trabecular pattern and increased cortical wall thickness. Bone histology and bone marrow derived macrophage cultures in these mice revealed increased osteoclastogenesis observed by TRAP staining suggestive of Paget bone disease. The VCP(R155H/+ knock-in mice replicate the muscle, bone and brain pathology of inclusion body myopathy, thus representing a useful model for preclinical studies.

  12. The P2 of Wheat yellow mosaic virus rearranges the endoplasmic reticulum and recruits other viral proteins into replication-associated inclusion bodies.

    Science.gov (United States)

    Sun, Liying; Andika, Ida Bagus; Shen, Jiangfeng; Yang, Di; Chen, Jianping

    2014-06-01

    Viruses commonly modify host endomembranes to facilitate biological processes in the viral life cycle. Infection by viruses belonging to the genus Bymovirus (family Potyviridae) has long been known to induce the formation of large membranous inclusion bodies in host cells, but their assembly and biological roles are still unclear. Immunoelectron microscopy of cells infected with the bymovirus Wheat yellow mosaic virus (WYMV) showed that P1, P2 and P3 are the major viral protein constituents of the membranous inclusions, whereas NIa-Pro (nuclear inclusion-a protease) and VPg (viral protein genome-linked) are probable minor components. P1, P2 and P3 associated with the endoplasmic reticulum (ER), but only P2 was able to rearrange ER and form large aggregate structures. Bioinformatic analyses and chemical experiments showed that P2 is an integral membrane protein and depends on the active secretory pathway to form aggregates of ER membranes. In planta and in vitro assays demonstrated that P2 interacts with P1, P3, NIa-Pro or VPg and recruits these proteins into the aggregates. In vivo RNA labelling using WYMV-infected wheat protoplasts showed that the synthesis of viral RNAs occurs in the P2-associated inclusions. Our results suggest that P2 plays a major role in the formation of membranous compartments that house the genomic replication of WYMV. © 2013 BSPP AND JOHN WILEY & SONS LTD.

  13. NIH Human Microbiome Project defines normal bacterial makeup of the body

    Science.gov (United States)

    Microbes inhabit just about every part of the human body, living on the skin, in the gut, and up the nose. Sometimes they cause sickness, but most of the time, microorganisms live in harmony with their human hosts, providing vital functions essential for

  14. Correlations between cyanobacterial density and bacterial transformation to the viable but nonculturable (VBNC) state in four freshwater water bodies.

    Science.gov (United States)

    Chen, Huirong; Shen, Ju; Pan, Gaoshan; Liu, Jing; Li, Jiancheng; Hu, Zhangli

    2015-10-01

    Nutrient concentrations, phytoplankton density and community composition, and the viable but nonculturable (VBNC) state of heterotrophic bacteria were investigated in three connected reservoirs and a small isolated lake in South China to study the relationship between biotic and abiotic factors and the VBNC state in bacteria. Nutrient concentrations in the reservoirs increased in the direction of water flow, whereas Wenshan Lake was more eutrophic. Cyanobacterial blooms occurred in all four water bodies, with differing seasonal trends and dominant species. In Xili and Tiegang Reservoirs, the VBNC ratio (percent of VBNC state bacteria over total viable bacteria) was high for most of the year and negatively correlated with cyanobacterial density. Laboratory co-culture experiments were performed with four heterotrophic bacterial species isolated from Wenshan Lake (Escherichia coli, Klebsiella peneumoniae, Bacillus megaterium and Bacillus cereus) and the dominant cyanobacterial species (Microcystis aeruginosa). For the first three bacterial species, the presence of M. aeruginosa induced the VBNC state and the VBNC ratio was positively correlated with M. aeruginosa density. However, B. cereus inhibited M. aeruginosa growth. These results demonstrate that cyanobacteria could potentially regulate the transformation to the VBNC state of waterborne bacteria, and suggest a role for bacteria in cyanobacterial bloom initiation and termination.

  15. Constraining the Thermochronological History of the IAB Parent Body: High Resolution Ar-40-Ar-39 Ages on Plagioclase Separates from Silicate Inclusions of IAB Meteorites

    Science.gov (United States)

    Vogel, N.; Renne, P. R.

    2004-12-01

    The processes that led to the assembly of primitive inclusions in a once molten metal matrix as represented by IAB meteorites have not yet been fully understood [1]. Ar-Ar dating of the inclusions provides important information about the thermal history of the IAB parent body [e.g., 2, 3], but the analysis of bulk inclusions, the standard procedure in the past, is often impaired by excess 40Ar and redistribution or loss of K and/or Ar during the history of the meteoriod and in the reactor. To minimize these problems, we prepared from silicate inclusions of four IABs pure plagioclase separates of different grain sizes and quality grades. On these we performed high resolution stepwise Ar-40-Ar-39 dating. Preliminary ages for the different separates of the inclusions are, in Ma, 4540(11) to 4459(12) for Caddo County, 4500(20) to 4380(30) for Landes, 4440(50) to 4340(30) for Ocotillo, and 4480(40) to 4200(30) and 4430(30) to 4300(30) for CDC2 and CDC1, respectively. The age ranges might reflect the residence time of each inclusion in the K-Ar blocking temperature range (ca. 600 K), and is narrowest for Caddo County, being also the oldest inclusion studied by us. Assuming that IABs resulted from a collision of a molten metal body with a chondritic planetesimal [4], Caddo County could represent a surface sample explaining the early and fast cooling, whereas the other samples might have been buried deeper within the IAB body, subject to prolonged residence at elevated temperatures. If IABs formed in impact metal melt pools peppered with chondritic host material [5] the different cooling ages, and age ranges recorded in each inclusion could reflect residence times in a certain metal melt pool, which indirectly would translate into pool sizes and the energies released by the previous impacts. Also, there may have been more than one IAB parent body. Whatever process led to the formation of IAB meteorites was active already very early in the history of the solar system, in

  16. Human body temperature and new approaches to constructing temperature-sensitive bacterial vaccines.

    Science.gov (United States)

    White, Matthew D; Bosio, Catharine M; Duplantis, Barry N; Nano, Francis E

    2011-09-01

    Many of the live human and animal vaccines that are currently in use are attenuated by virtue of their temperature-sensitive (TS) replication. These vaccines are able to function because they can take advantage of sites in mammalian bodies that are cooler than the core temperature, where TS vaccines fail to replicate. In this article, we discuss the distribution of temperature in the human body, and relate how the temperature differential can be exploited for designing and using TS vaccines. We also examine how one of the coolest organs of the body, the skin, contains antigen-processing cells that can be targeted to provoke the desired immune response from a TS vaccine. We describe traditional approaches to making TS vaccines, and highlight new information and technologies that are being used to create a new generation of engineered TS vaccines. We pay particular attention to the recently described technology of substituting essential genes from Arctic bacteria for their homologues in mammalian pathogens as a way of creating TS vaccines.

  17. Effects of municipal sewage on the sessile and planctonic bacterial flora of bodies of surface water

    International Nuclear Information System (INIS)

    Lorch, H.J.; Ottow, J.C.G.

    1992-01-01

    For the evaluation of the useability of bacteria as biological indicators of the water state, sections of a flowing body of water (Erms, Schwaebische Alb) with different pollutant levels were investigated between August '82 and July '85. In flowing waves (current biological indicators), the population densities of saprophytes, coliform buds and sporophytes were determined as well as the total number of bacteria number and the morphology of the bacteria. The upgrowth on artificial (glass) and natural (water plants) substrates (integrating biological indicators) was examined with the aid of light microscope and scanning electron microscope in order to measure the long-term influence of different nutrient conditions on the microflora. (orig.) [de

  18. The bacterial communities associated with fecal types and body weight of rex rabbits

    OpenAIRE

    Bo Zeng; Shushu Han; Ping Wang; Bin Wen; Wensu Jian; Wei Guo; Zhiju Yu; Dan Du; Xiangchao Fu; Fanli Kong; Mingyao Yang; Xiaohui Si; Jiangchao Zhao; Ying Li

    2015-01-01

    Rex rabbit is an important small herbivore for fur and meat production. However, little is known about the gut microbiota in rex rabbit, especially regarding their relationship with different fecal types and growth of the hosts. We characterized the microbiota of both hard and soft feces from rex rabbits with high and low body weight by using the Illumina MiSeq platform targeting the V4 region of the 16S rDNA. High weight rex rabbits possess distinctive microbiota in hard feces, but not in so...

  19. Development of a rapid high-efficiency scalable process for acetylated Sus scrofa cationic trypsin production from Escherichia coli inclusion bodies.

    Science.gov (United States)

    Zhao, Mingzhi; Wu, Feilin; Xu, Ping

    2015-12-01

    Trypsin is one of the most important enzymatic tools in proteomics and biopharmaceutical studies. Here, we describe the complete recombinant expression and purification from a trypsinogen expression vector construct. The Sus scrofa cationic trypsin gene with a propeptide sequence was optimized according to Escherichia coli codon-usage bias and chemically synthesized. The gene was inserted into pET-11c plasmid to yield an expression vector. Using high-density E. coli fed-batch fermentation, trypsinogen was expressed in inclusion bodies at 1.47 g/L. The inclusion body was refolded with a high yield of 36%. The purified trypsinogen was then activated to produce trypsin. To address stability problems, the trypsin thus produced was acetylated. The final product was generated upon gel filtration. The final yield of acetylated trypsin was 182 mg/L from a 5-L fermenter. Our acetylated trypsin product demonstrated higher BAEE activity (30,100 BAEE unit/mg) than a commercial product (9500 BAEE unit/mg, Promega). It also demonstrated resistance to autolysis. This is the first report of production of acetylated recombinant trypsin that is stable and suitable for scale-up. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Expression, characterization of a novel nitrilase PpL19 from Pseudomonas psychrotolerans with S-selectivity toward mandelonitrile present in active inclusion bodies.

    Science.gov (United States)

    Sun, Huihui; Gao, Wenyuan; Wang, Hualei; Wei, Dongzhi

    2016-03-01

    To identify a novel nitrilase with S-selectivity toward mandelonitrile that can produce (S)-mandelic acid in one step. A novel nitrilase PpL19 from Pseudomonas psychrotolerans L19 was discovered by genome mining. It showed S-selectivity with an enantiomeric excess of 52.7 % when used to hydrolyse (R, S)-mandelonitrile. No byproduct was observed. PpL19 was overexpressed in Escherichia coli BL21 (DE3) and formed inclusion bodies that were active toward mandelonitrile and stable across a broad range of temperature and pH. In addition, PpL19 hydrolysed nitriles with diverse structures; arylacetonitriles were the optimal substrates. Homology modelling and docking studies of both enantiomers of mandelonitrile in the active site of nitrilase PpL19 shed light on the enantioselectivity. A novel nitrilase PpL19 from P. psychrotolerans L19 was mined and distinguished from other nitrilases as it was expressed as an active inclusion body and showed S-selectivity toward mandelonitrile.

  1. Continuous processing of recombinant proteins: Integration of inclusion body solubilization and refolding using simulated moving bed size exclusion chromatography with buffer recycling.

    Science.gov (United States)

    Wellhoefer, Martin; Sprinzl, Wolfgang; Hahn, Rainer; Jungbauer, Alois

    2013-12-06

    An integrated process which combines continuous inclusion body dissolution with NaOH and continuous matrix-assisted refolding based on closed-loop simulated moving bed size exclusion chromatography was designed and experimentally evaluated at laboratory scale. Inclusion bodies from N(pro) fusion pep6His and N(pro) fusion MCP1 from high cell density fermentation were continuously dissolved with NaOH, filtered and mixed with concentrated refolding buffer prior to refolding by size exclusion chromatography (SEC). This process enabled an isocratic operation of the simulated moving bed (SMB) system with a closed-loop set-up with refolding buffer as the desorbent buffer and buffer recycling by concentrating the raffinate using tangential flow filtration. With this continuous refolding process, we increased the refolding and cleavage yield of both model proteins by 10% compared to batch dilution refolding. Furthermore, more than 99% of the refolding buffer of the raffinate could be recycled which reduced the buffer consumption significantly. Based on the actual refolding data, we compared throughput, productivity, and buffer consumption between two batch dilution refolding processes - one using urea for IB dissolution, the other one using NaOH for IB dissolution - and our continuous refolding process. The higher complexity of the continuous refolding process was rewarded with higher throughput and productivity as well as significantly lower buffer consumption compared to the batch dilution refolding processes. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Effects of inclusion levels of Indigofera sp. on feed intake, digestibility and body weight gain in kids fed Brachiaria ruziziensis.

    Directory of Open Access Journals (Sweden)

    Andi Tarigan

    2011-03-01

    Full Text Available Twenty weaned male goats (F1 of Boer x Kacang with initial weight of 9 to 12 kg and ages ranging between 3.0 and 4.0 months were used in a study to evaluate the increasing inclusion of Indigofera sp foliage as a source of protein in diets based on chopped Brachiaria ruziziensis for growing goats. Five goats were allocated to one of four treatments in a randomised block design. The diet treatments were: T0 (control diets: B. ruziziensis (100%, T1 (85% B. ruziziensis + 15% Indigofera sp., T2 (70% B. ruziziensis + 30% Indigofera sp. T3 (55% B. ruziziensis + 45% Indigofera sp. all on DM basis. Feed (DM was offered daily at 3.5% BW. The content of CP in Indigofera sp is relatively high (258 g/kg DM, while the NDF (350.7 g/kg DM and ADF (232.2 g/ kg DM concentrations were low. The content of secondary compounds such as total phenol (8.9 g/kg DM, total tannin (0.8 g/kg DM and condensed tannin (0,5 g/kg DM were considerably low. The inclusion of Indigofera sp foliage in diets increased (P 0.05 among the T0,T1 and T2 diets and ranged from 0.08 to 0.09. It is concluded that the foliage of Indigofera sp could be used as feed supplement to supply proteins with low tannin contents. In a grass-based diets Indigofera sp colud be used at the level of 30 to 45% (DM for growing kids.

  3. Inclusion of Body-Bias Effect in SPICE Modeling of 4H-SiC Integrated Circuit Resistors

    Science.gov (United States)

    Neudeck, Philip G.

    2017-01-01

    The DC electrical behavior of n-type 4H-SiC resistors used for realizing 500 C durable integrated circuits (ICs) is studied as a function of substrate bias and temperature. Improved fidelity electrical simulation is described using SPICE NMOS model to simulate resistor substrate body bias effect that is absent from the SPICE semiconductor resistor model.

  4. Inclusion of Body Bias Effect in SPICE Modeling of 4H-SiC Integrated Circuit Resistors

    Science.gov (United States)

    Neudeck, Philip G.

    2017-01-01

    The DC electrical behavior of n-type 4H-SiC resistors used for realizing 500 degrees Celsius durable integrated circuits (ICs) is studied as a function of substrate bias and temperature. Improved fidelity electrical simulation is described using SPICE NMOS model to simulate resistor substrate body bias effect that is absent from the SPICE semiconductor resistor model.

  5. Whole-body MRI in patients with Non-bacterial Osteitis: Radiological findings and correlation with clinical data

    Energy Technology Data Exchange (ETDEWEB)

    Arnoldi, A.P.; Geyer, L.L. [Institute for Clinical Radiology, Ludwig-Maximilians-University Hospital, Munich (Germany); Schlett, C.L.; Weckbach, S. [University Hospital Heidelberg, Diagnostic and Interventional Radiology, Heidelberg (Germany); Douis, H. [University Hospital Birmingham, Department of Radiology, Birmingham (United Kingdom); Voit, A.M.; Bleisteiner, F.; Jansson, A.F. [Dr. von Hauner Children' s Hospital, Ludwig-Maximilians-University, Department of Rheumatology and Immunology, Munich (Germany)

    2017-06-15

    To correlate clinical findings of Non-bacterial Osteitis (NBO) with whole-body MRI (WB-MRI) findings and determine a radiologic index for NBO (RINBO) which allows standardized reporting of WB-MRI. In a prospective study, 40 patients with diagnosis of NBO underwent clinical examination and WB-MRI in which STIR- and T1- weighted images were assessed for NBO-typical lesions. Parameters of interest for RINBO were: number of radiologically active lesions (RAL), size of the patients' maximum RAL presence of extramedullary and spinal involvement. Results were tested for statistical agreement of clinical and MR-based lesion detection. RINBO was tested for correlation with clinical activity. 62/95 clinically/radiologically active lesions were found in 30/33 patients. In 45 % of the cohort, more active lesions were detected by WB-MRI than by clinical examination. RINBO was a significant predictor for the presence of clinically active lesions. WB-MRI is a powerful diagnostic tool for patients with NBO which can reveal asymptomatic disease activity. With RINBO a standardized evaluation approach is proposed which helps assessing radiologic disease burden and predicts clinical disease activity. (orig.)

  6. Purification and partial genome characterization of the bacterial endosymbiont Blattabacterium cuenoti from the fat bodies of cockroaches

    Directory of Open Access Journals (Sweden)

    Yamada Akinori

    2008-11-01

    Full Text Available Abstract Background Symbiotic relationships between intracellular bacteria and eukaryotes are widespread in nature. Genome sequencing of the bacterial partner has provided a number of key insights into the basis of these symbioses. A challenging aspect of sequencing symbiont genomes is separating the bacteria from the host tissues. In the present study, we describe a simple method of endosymbiont purification from complex environment, using Blattabacterium cuenoti inhabiting in cockroaches as a model system. Findings B. cuenoti cells were successfully purified from the fat bodies of the cockroach Panesthia angustipennis by a combination of slow- and fast-speed centrifugal fractionations, nylon-membrane filtration, and centrifugation with Percoll solutions. We performed pulse-field electrophoresis, diagnostic PCR and random sequencing of the shoutgun library. These experiments confirmed minimal contamination of host and mitochondrial DNA. The genome size and the G+C content of B. cuenoti were inferred to be 650 kb and 32.1 ± 7.6%, respectively. Conclusion The present study showed successful purification and characterization of the genome of B. cuenoti. Our methodology should be applicable for future symbiont genome sequencing projects. An advantage of the present purification method is that each step is easily performed with ordinary microtubes and a microcentrifuge, and without DNase treatment.

  7. Effects of blood-flow-restricted resistance training on muscle function in a 74-year-old male with sporadic inclusion body myositis

    DEFF Research Database (Denmark)

    Jørgensen, Anders Nørkær; Aagaard, P; Nielsen, J L

    2016-01-01

    Sporadic inclusion body myositis (sIBM) is a systemic disease that is characterized by substantial skeletal muscle weakness and muscle inflammation, leading to impaired physical function. The objective was to investigate the effect of low-load resistance exercise with concurrent partial blood flow...... restriction to the working muscles (blood-flow-restricted (BFR) training) in a patient with sIBM. The training consisted of 12 weeks of lower extremity BFR training with low training loads (~25-RM). The patient was tested for mechanical muscle function and functional capacity before and after 6 and 12 weeks...... of training. Maximal horizontal gait speed increased by 19%, which was accompanied by 38-92% improvements in mechanical muscle function (maximal isometric strength, rate of force development and muscle power). In conclusion, BFR training was well tolerated by the patient with sIBM and led to substantial...

  8. Obtaining Soluble Folded Proteins from Inclusion Bodies Using Sarkosyl, Triton X-100, and CHAPS: Application to LB and M9 Minimal Media.

    Science.gov (United States)

    Massiah, Michael A; Wright, Katharine M; Du, Haijuan

    2016-04-01

    This unit describes a straightforward and efficient method of using sarkosyl to solubilize and recover difficult recombinant proteins, such as GST- and His6 -tagged fusion proteins, that are overexpressed in E. coli. This protocol is especially useful for rescuing recombinant proteins overexpressed in M9 minimal medium. Sarkosyl added to lysis buffers helps with both protein solubility and cell lysis. Higher percentage sarkosyl (up to 10%) can extract >95% of soluble protein from inclusion bodies. In the case of sarkosyl-solubilized GST-fusion proteins, batch-mode affinity purification requires addition of a specific ratio of Triton X-100 and CHAPS, while sarkosyl-solubilized His6 -tagged fusion proteins can be directly purified on Ni(2+) resin columns. Proteins purified by this method could be widely used in biological assays, structure analysis and mass spectrum assay. Copyright © 2016 John Wiley & Sons, Inc.

  9. Purification and refolding of anti-T-antigen single chain antibodies (scFvs) expressed in Escherichia coli as inclusion bodies.

    Science.gov (United States)

    Yuasa, Noriyuki; Koyama, Tsubasa; Fujita-Yamaguchi, Yoko

    2014-02-01

    T-antigen (Galβ1-3GalNAcα-1-Ser/Thr) is an oncofetal antigen that is commonly expressed as a carbohydrate determinant in many adenocarcinomas. Since it is associated with tumor progression and metastasis, production of recombinant antibodies specific for T-antigen could lead to the development of cancer diagnostics and therapeutics. Previously, we isolated and characterized 11 anti-T-antigen phage clones from a phage library displaying human single-chain antibodies (scFvs) and purified one scFv protein, 1G11. More recently, we purified and characterized 1E8 scFv protein using a Drosophila S2 expression system. In the current study, four anti-T-antigen scFv genes belonging to Groups 1-4 were purified from inclusion bodies expressed in Escherichia coli cells. Inclusion bodies isolated from E. coli cells were denatured in 3.5 M Gdn-HCl. Solubilized His-tagged scFv proteins were purified using Ni(2+)-Sepharose column chromatography in the presence of 3.5 M Gdn-HCl. Purified scFv proteins were refolded according to a previously published method of step-wise dialysis. Two anti-T-antigen scFv proteins, 1E6 and 1E8 that belong to Groups 1 and 2, respectively, were produced in sufficient amounts, thus allowing further characterization of their binding activity with T-antigen. Specificity and affinity constants determined using enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR), respectively, provided evidence that both 1E8 and 1E6 scFv proteins are T-antigen specific and suggested that 1E8 scFv protein has a higher affinity for T-antigen than 1E6 scFv protein.

  10. A Drosophila model of dominant inclusion body myopathy type 3 shows diminished myosin kinetics that reduce muscle power and yield myofibrillar defects.

    Science.gov (United States)

    Suggs, Jennifer A; Melkani, Girish C; Glasheen, Bernadette M; Detor, Mia M; Melkani, Anju; Marsan, Nathan P; Swank, Douglas M; Bernstein, Sanford I

    2017-06-01

    Individuals with inclusion body myopathy type 3 (IBM3) display congenital joint contractures with early-onset muscle weakness that becomes more severe in adulthood. The disease arises from an autosomal dominant point mutation causing an E706K substitution in myosin heavy chain type IIa. We have previously expressed the corresponding myosin mutation (E701K) in homozygous Drosophila indirect flight muscles and recapitulated the myofibrillar degeneration and inclusion bodies observed in the human disease. We have also found that purified E701K myosin has dramatically reduced actin-sliding velocity and ATPase levels. Since IBM3 is a dominant condition, we now examine the disease state in heterozygote Drosophila in order to gain a mechanistic understanding of E701K pathogenicity. Myosin ATPase activities in heterozygotes suggest that approximately equimolar levels of myosin accumulate from each allele. In vitro actin sliding velocity rates for myosin isolated from the heterozygotes were lower than the control, but higher than for the pure mutant isoform. Although sarcomeric ultrastructure was nearly wild type in young adults, mechanical analysis of skinned indirect flight muscle fibers revealed a 59% decrease in maximum oscillatory power generation and an approximately 20% reduction in the frequency at which maximum power was produced. Rate constant analyses suggest a decrease in the rate of myosin attachment to actin, with myosin spending decreased time in the strongly bound state. These mechanical alterations result in a one-third decrease in wing beat frequency and marginal flight ability. With aging, muscle ultrastructure and function progressively declined. Aged myofibrils showed Z-line streaming, consistent with the human heterozygote phenotype. Based upon the mechanical studies, we hypothesize that the mutation decreases the probability of the power stroke occurring and/or alters the degree of movement of the myosin lever arm, resulting in decreased in vitro

  11. A Drosophila model of dominant inclusion body myopathy type 3 shows diminished myosin kinetics that reduce muscle power and yield myofibrillar defects

    Directory of Open Access Journals (Sweden)

    Jennifer A. Suggs

    2017-06-01

    Full Text Available Individuals with inclusion body myopathy type 3 (IBM3 display congenital joint contractures with early-onset muscle weakness that becomes more severe in adulthood. The disease arises from an autosomal dominant point mutation causing an E706K substitution in myosin heavy chain type IIa. We have previously expressed the corresponding myosin mutation (E701K in homozygous Drosophila indirect flight muscles and recapitulated the myofibrillar degeneration and inclusion bodies observed in the human disease. We have also found that purified E701K myosin has dramatically reduced actin-sliding velocity and ATPase levels. Since IBM3 is a dominant condition, we now examine the disease state in heterozygote Drosophila in order to gain a mechanistic understanding of E701K pathogenicity. Myosin ATPase activities in heterozygotes suggest that approximately equimolar levels of myosin accumulate from each allele. In vitro actin sliding velocity rates for myosin isolated from the heterozygotes were lower than the control, but higher than for the pure mutant isoform. Although sarcomeric ultrastructure was nearly wild type in young adults, mechanical analysis of skinned indirect flight muscle fibers revealed a 59% decrease in maximum oscillatory power generation and an approximately 20% reduction in the frequency at which maximum power was produced. Rate constant analyses suggest a decrease in the rate of myosin attachment to actin, with myosin spending decreased time in the strongly bound state. These mechanical alterations result in a one-third decrease in wing beat frequency and marginal flight ability. With aging, muscle ultrastructure and function progressively declined. Aged myofibrils showed Z-line streaming, consistent with the human heterozygote phenotype. Based upon the mechanical studies, we hypothesize that the mutation decreases the probability of the power stroke occurring and/or alters the degree of movement of the myosin lever arm, resulting in

  12. Effects of exposure to thin-ideal media images on body dissatisfaction: testing the inclusion of a disclaimer versus warning label.

    Science.gov (United States)

    Ata, Rheanna N; Thompson, J Kevin; Small, Brent J

    2013-09-01

    The current study was designed to determine whether the inclusion of a disclaimer (i.e., "Retouched photograph aimed at changing a person's physical appearance.") or warning (i.e., "Warning: Trying to look as thin as this model may be dangerous to your health.") added to images of thin/attractive models would affect body dissatisfaction and intent to diet in female undergraduate students (n=342). Participants were randomly assigned to one of four groups: (a) disclaimer, (b) warning, (c) model control, or (d) car control. Results revealed a significant interaction between group and time, whereby only the car control group reported a significant change (i.e., decrease) in body dissatisfaction over time. Groups did not differ on intent to diet measured at post-exposure. The results largely replicate other findings in this area and call into question advocacy efforts to label media images as a strategy to decrease women's identification with the stimuli. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. An in-depth characterization of the entomopathogenic strain Bacillus pumilus 15.1 reveals that it produces inclusion bodies similar to the parasporal crystals of Bacillus thuringiensis.

    Science.gov (United States)

    Garcia-Ramon, Diana C; Molina, C Alfonso; Osuna, Antonio; Vílchez, Susana

    2016-04-01

    In the present work, the local isolate Bacillus pumilus 15.1 has been morphologically and biochemically characterized in order to gain a better understanding of this novel entomopathogenic strain active against Ceratitis capitata. This strain could represent an interesting biothechnological tool for the control of this pest. Here, we report on its nutrient preferences, extracellular enzyme production, motility mechanism, biofilm production, antibiotic suceptibility, natural resistance to chemical and physical insults, and morphology of the vegetative cells and spores. The pathogen was found to be β-hemolytic and susceptible to penicillin, ampicillin, chloramphenicol, gentamicin, kanamycin, rifampicin, tetracycline, and streptomycin. We also report a series of biocide, thermal, and UV treatments that reduce the viability of B. pumilus 15.1 by several orders of magnitude. Heat and chemical treatments kill at least 99.9 % of vegetative cells, but spores were much more resistant. Bleach was the only chemical that was able to completely eliminate B. pumilus 15.1 spores. Compared to the B. subtilis 168 spores, B. pumilus 15.1 spores were between 2.67 and 350 times more resistant to UV radiation while the vegetative cells of B. pumilus 15.1 were almost up to 3 orders of magnitude more resistant than the model strain. We performed electron microscopy for morphological characterization, and we observed geometric structures resembling the parasporal crystal inclusions synthesized by Bacillus thuringiensis. Some of the results obtained here such as the parasporal inclusion bodies produced by B. pumilus 15.1 could potentially represent virulence factors of this novel and potentially interesting strain.

  14. Frequency and circumstances of falls in people with inclusion body myositis: a questionnaire survey to explore falls management and physiotherapy provision.

    Science.gov (United States)

    Hiscock, A; Dewar, L; Parton, M; Machado, P; Hanna, M; Ramdharry, G

    2014-03-01

    To survey the incidence and circumstances of falls for people with inclusion body myositis (IBM) in the UK, and to investigate the provision of physiotherapy and falls management. Postal questionnaire survey. Participants completed questionnaires at home. Ninety-four people diagnosed with IBM were screened against the inclusion criteria. Seventy-two potential participants were sent a questionnaire, and 62 were completed and returned. Invited participants were sent an adapted Falls Event Questionnaire pertaining to falls, perceived causes of falls and the provision of physiotherapy. Questionnaires were returned anonymously. The proportions of respondents who reported a fall or a near fall, along with the frequencies of falls and near falls were calculated. Descriptive data of falls were collected pertaining to location and cause. Data analysis was performed to investigate provision of physiotherapy services. The response rate was 86% [62/72, mean (standard deviation) age 68 (8) years]. Falls were reported by 98% (61/62) of respondents, with 60% (37/62) falling frequently. In this study, age was not found to be an indicator of falls risk or frequency. Twenty-one percent (13/62) of respondents had not seen a physiotherapist in relation to their IBM symptoms, and of those that had, 31% (15/49) had not seen a physiotherapist until more than 12 months after IBM was diagnosed. Only 18% (11/61) of fallers reported that they had received falls management input. Falls are a common occurrence for people with IBM, independent of age and years since symptoms first presented, and are poorly addressed by appropriate physiotherapy management. National falls guidelines are not being followed, and referral rates to physiotherapy need to improve. Copyright © 2013 Chartered Society of Physiotherapy. Published by Elsevier Ltd. All rights reserved.

  15. Inclusive pedagogy

    DEFF Research Database (Denmark)

    Korsgaard, Morten Timmermann; Mortensen, Stig Skov

    With a starting point in the tradition of geisteswissenschaftliche Pädagogik this article presents a challenge to inclusive education research to engage a continental perspective on educational research. The motivation is to entice inclusive education researchers to begin to ask educational...... questions of inclusion, as opposed to inclusive questions of education. Recent years has seen a call to re-think inclusive education research and this paper attempts to answer this call by turning to a Continental perspective and the emphasis on an at least relative autonomy for the theory and practice...... and the politicisation of inclusive education, and a positive aim in the form of an argument for a move towards constructing a pedagogical ideal of inclusion....

  16. Inclusion body hepatitis associated with an outbreak of fowl adenovirus type 2 and type 8b in broiler flocks in South Africa

    Directory of Open Access Journals (Sweden)

    Louis H. Maartens

    2014-12-01

    Full Text Available Inclusion body hepatitis is an acute disease of chickens ascribed to viruses of the genus Aviadenovirus and referred to as fowl adenovirus (FAdV. There are 12 FAdV types (FAdV1to FAdV8a and FAdV8b to FAdV11, classified into five species based on their genotype (designated FAdVA to FAdVE. A total of 218 000 chickens, 2–29 days of age, were affected over a 1-year period, all testing positive by microscopy, virus isolation and confirmation with polymerase chain reaction (PCR. Affected birds were depressed, lost body weight,were weak and had watery droppings. Pathological changes observed during necropsy indicated consistent changes in the liver, characterised by hepatomegaly, cholestasis and hepatitis. Lesions were also discernible in the spleen, kidney and gizzard wall and were characterised by splenomegaly, pinpoint haemorrhages, nephritis with haemorrhage,visceral gout and serosal ecchymosis of the gizzard wall. Histopathological lesions were most consistently observed in the liver but could also be seen in renal and splenic tissue. Virus isolation was achieved in embryonated eggs and most embryos revealed multifocalto diffuse hepatic necrosis, with a mixed cellular infiltrate of macrophages and heterophils(necro-granulomas, even in the absence of macroscopic pathology. Virus isolation results were verified by histopathology and PCR on embryonic material and further characterised by nucleotide sequence analysis. Two infectious bursal disease virus isolates were also made from the Klerksdorp flock. Nucleotide sequence analysis of the L1 hexon loop of all the FAdV isolates indicated homology (99% with prototype strains P7-A for FAdV-2, as well as for FAdV-8b.

  17. Sporadic-inclusion body myositis (s-IBM) is not so prevalent in Istanbul/Turkey: a muscle biopsy based survey.

    Science.gov (United States)

    Oflazer, P Serdaroglu; Deymeer, F; Parman, Y

    2011-06-01

    In a muscle biopsy based study, only 9 out of 5450 biopsy samples, received from all parts of greater Istanbul area, had typical clinical and most suggestive light microscopic sporadic-inclusion body myositis (s-IBM) findings. Two other patients with and ten further patients without characteristic light microscopic findings had referring diagnosis of s-IBM. As the general and the age-adjusted populations of Istanbul in 2010 were 13.255.685 and 2.347.300 respectively, the calculated corresponding 'estimated prevalences' of most suggestive s-IBM in the Istanbul area were 0.679 X 10(-6) and 3.834 X 10(-6). Since Istanbul receives heavy migration from all regions of Turkey and ours is the only muscle pathology laboratory in Istanbul, projection of these figures to the Turkish population was considered to be reasonable and an estimate of the prevalence of s-IBM in Turkey was obtained. The calculated 'estimated prevalence' of s-IBM in Turkey is lower than the previously reported rates from other countries. The wide variation in the prevalence rates of s-IBM may reflect different genetic, immunogenetic or environmental factors in different populations.

  18. Body

    OpenAIRE

    Riggs, Christina

    2010-01-01

    The human body is both the physical form inhabited by an individual “self” and the medium through which an individual engages with society. Hence the body both shapes and is shaped by an individual’s social roles. In contrast to the cognate fields of archaeology, anthropology, and classics, there has been little explicit discussion or theorization of the body in Egyptology. Some recent works, discussed here, constitute an exception to this trend, but there is much more scope for exploring anc...

  19. Inclusive pedagogy

    DEFF Research Database (Denmark)

    Korsgaard, Morten Timmermann; Skov Mortensen, Stig

    This article will present a case for a shift in perspective in inclusive education research towards a continentally inspired approach. Drawing on the age old distinction between continental and Anglo-American educational research the aim is to flesh out what a shift to a continental approach...... will entail, and why it might be beneficial to research in inclusive education...

  20. Social inclusion and inclusive education

    Directory of Open Access Journals (Sweden)

    Marsela Robo

    2014-07-01

    In line with global debate on social inclusion and exclusion, the author brings the way this debate has now pervaded both the official and development policy discourse in Albania.Social inclusion is considered as one of the priorities of the current government, with poverty reduction as its main focus, which will be ensured not only through economic development. In the end, the article focuses on the role of education as a very important and useful tool for ensuring social inclusion.Social inclusion through education, in particular through vocational education, considered by the author as the only way towards sustainable development of Albanian society.

  1. Sequential tentacle grafting and charge modification for enhancing charge density of mono-sized beads for facilitated protein refolding and purification from inclusion bodies.

    Science.gov (United States)

    Dong, Xiao-Yan; Chen, Ran; Yang, Chun-Yan; Sun, Yan

    2014-06-20

    We have previously found that addition of like-charged media in a refolding solution can greatly enhance the refolding of pure proteins by suppressing protein aggregation. Herein, negatively charged mono-sized microspheres with sulfonic groups were fabricated to explore the facilitating effect of like-charged media on the refolding of enhanced green fluorescent protein (EGFP) expressed as inclusion bodies (IBs). A sequential polymer-tentacle grafting and sulfonate modification strategy was developed to increase the charge density of mono-sized poly(glycidyl methacrylate) (pGMA) beads (2.4μm). Namely, GMA was first grafted onto the beads by grafting polymerization to form poly(GMA) tentacles on the pGMA beads, and then the epoxy groups on the tentacles were converted into sulfonic groups by modification with sodium sulfite. By this fabrication strategy, the charge density of the beads reached 793μmol/g, about 2.8 times higher than that modified without prior grafting of the pGMA beads (285μmol/g). The negatively charged beads of different charge densities were used for facilitating the refolding of like-charged EGFP from IBs. The refolding yield as well as refolding rate increased with increasing charge density. The anti-aggregation effects of urea and like-charged microspheres were synergetic. In addition, partial purification of EGFP was achieved because the ion-exchange adsorption led to 52% removal of positively charged contaminant proteins in the refolded solution. Finally, reusability of the tentacle beads was demonstrated by repetitive EGFP refolding and recovery cycles. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Inclusion bodies of recombinant Epstein-Barr virus capsid antigen p18 as potential immobilized antigens in enzyme immunoassays for detection of nasopharyngeal carcinoma.

    Science.gov (United States)

    Lim, Chun Shen; Goh, Siang Ling; Kariapper, Leena; Krishnan, Gopala; Lim, Yat-Yuen; Ng, Ching Ching

    2015-08-25

    Development of indirect enzyme-linked immunosorbent assays (ELISAs) often utilizes synthetic peptides or recombinant proteins from Escherichia coli as immobilized antigens. Because inclusion bodies (IBs) formed during recombinant protein expression in E. coli are commonly thought as misfolded aggregates, only refolded proteins from IBs are used to develop new or in-house diagnostic assays. However, the promising utilities of IBs as nanomaterials and immobilized enzymes as shown in recent studies have led us to explore the potential use of IBs of recombinant Epstein-Barr virus viral capsid antigen p18 (VCA p18) as immobilized antigens in ELISAs for serologic detection of nasopharyngeal carcinoma (NPC). Thioredoxin fusion VCA p18 (VCA-Trx) and IBs of VCA p18 without fusion tags (VCA-IBs) were purified from E. coli. The diagnostic performances of IgG/VCA-IBs, IgG/VCA-Denat-IBs (using VCA-IBs coated in 8mol/l urea), IgG/VCA-Trx, and IgG/VCA-Peptide assays were compared by screening 100 NPC case-control pairs. The IgG/VCA-Denat-IBs assay showed the best area under the receiver operating characteristic curve (AUC: 0.802; p<0.05), while the AUCs for the IgG/VCA-IBs, IgG/VCA-Trx, and IgG/VCA-Peptide assays were comparable (AUC: 0.740, 0.727, and 0.741, respectively). We improved the diagnostic performance of the ELISA significantly using IBs of recombinant VCA p18. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Nuclear inclusion bodies of mutant and wild-type p53 in cancer: a hallmark of p53 inactivation and proteostasis remodelling by p53 aggregation.

    Science.gov (United States)

    De Smet, Frederik; Saiz Rubio, Mirian; Hompes, Daphne; Naus, Evelyne; De Baets, Greet; Langenberg, Tobias; Hipp, Mark S; Houben, Bert; Claes, Filip; Charbonneau, Sarah; Delgado Blanco, Javier; Plaisance, Stephane; Ramkissoon, Shakti; Ramkissoon, Lori; Simons, Colinda; van den Brandt, Piet; Weijenberg, Matty; Van England, Manon; Lambrechts, Sandrina; Amant, Frederic; D'Hoore, André; Ligon, Keith L; Sagaert, Xavier; Schymkowitz, Joost; Rousseau, Frederic

    2017-05-01

    Although p53 protein aggregates have been observed in cancer cell lines and tumour tissue, their impact in cancer remains largely unknown. Here, we extensively screened for p53 aggregation phenotypes in tumour biopsies, and identified nuclear inclusion bodies (nIBs) of transcriptionally inactive mutant or wild-type p53 as the most frequent aggregation-like phenotype across six different cancer types. p53-positive nIBs co-stained with nuclear aggregation markers, and shared molecular hallmarks of nIBs commonly found in neurodegenerative disorders. In cell culture, tumour-associated stress was a strong inducer of p53 aggregation and nIB formation. This was most prominent for mutant p53, but could also be observed in wild-type p53 cell lines, for which nIB formation correlated with the loss of p53's transcriptional activity. Importantly, protein aggregation also fuelled the dysregulation of the proteostasis network in the tumour cell by inducing a hyperactivated, oncogenic heat-shock response, to which tumours are commonly addicted, and by overloading the proteasomal degradation system, an observation that was most pronounced for structurally destabilized mutant p53. Patients showing tumours with p53-positive nIBs suffered from a poor clinical outcome, similar to those with loss of p53 expression, and tumour biopsies showed a differential proteostatic expression profile associated with p53-positive nIBs. p53-positive nIBs therefore highlight a malignant state of the tumour that results from the interplay between (1) the functional inactivation of p53 through mutation and/or aggregation, and (2) microenvironmental stress, a combination that catalyses proteostatic dysregulation. This study highlights several unexpected clinical, biological and therapeutically unexplored parallels between cancer and neurodegeneration. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. Copyright © 2016 Pathological Society of Great

  4. Expression and purification of a novel therapeutic single-chain variable fragment antibody against BNP from inclusion bodies of Escherichia coli.

    Science.gov (United States)

    Bu, Dawei; Zhou, Yuwei; Tang, Jian; Jing, Fang; Zhang, Wei

    2013-12-01

    Abnormal brain natriuretic peptide (BNP) secretion is regarded as the dominating mechanism of cerebral salt wasting syndrome (CSW), which results from a renal loss of sodium and water during intracranial disease leading to hyponatremia. Scale preparation of therapeutic single-chain variable fragment (scFv) that can neutralize elevated circulating BNP may have potential value for clinical use. In this report, we used a recently isolated humanized anti-BNP scFv fragment (3C1) as model antibody (Ab) to evaluate the potential of scale production of this therapeutic protein. The truncated gene encoding for scFv fragment cloned in pET22b (+) was mainly overexpressed as inclusion bodies in Escherichia coli (E. coli) Rosetta (DE3) pLysS cells. The insoluble fragment was solubilized and purified by Ni-NTA agarose resin under denaturation conditions, and recovered via an effective refolding buffer containing 50 mM Tris-HCl, pH 8.0, 0.15 M NaCl, 1 mM EDTA, 0.5 M arginine, 2 mM GSH, 1 mM GSSG, and 5% glycerol. The refolded scFv fragment was concentrated by PEG20000, and dialyzed in PBS (containing 5% glycerol, pH 7.4). The final yield was approximately 10.2 mg active scFv fragment per liter of culture (3.4 g wet weight cells). The scFv fragment was more than 95% pure assessed by SDS-PAGE assay. Recombinant scFv fragment with His tag displayed its immunoreactivity with anti-His tag Ab by western blotting. ELISA showed the scFv fragment specifically bound to BNP, and it displayed similar activity as the traditional anti-BNP monoclonal Ab (mAb). Thus, the current strategy allows convenient small-scale production of this therapeutic protein. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Adjuvant-enhanced antibody and cellular responses to inclusion bodies expressing FhSAP2 correlates with protection of mice to Fasciola hepatica.

    Science.gov (United States)

    Rivera, Francheska; Espino, Ana M

    2016-01-01

    Fasciola hepatica saposin-like protein-2 (FhSAP2) is a protein differentially expressed in various developmental stages of F. hepatica. Recombinant FhSAP2 has demonstrated the induction of partial protection in mice and rabbits when it is administered subcutaneously (SC) in Freund's adjuvant. Because FhSAP2 is overexpressed in bacteria in the form of inclusion bodies (IBs), we isolated IBs expressing FhSAP2 and tested their immunogenicity when administered SC in mice emulsified in two different adjuvants: QS-21 and Montanide TM ISA720. Animals received three injections containing 20 μg of protein two weeks apart and 4 weeks after the third injection, mice were infected with 10 F. hepatica metacercariae by oral route. The percentages of protection induced by FhSAP2-IBs were estimated to be between 60.0 and 62.5% when compared with adjuvant-vaccinated, infected controls. By determining the levels of IgG1 and IgG2a antibodies and IL-4 and IFNγ cytokines in the serum of experimental animals, it was found that both Th1 and Th2 immune responses were significantly increased in the FhSAP2-IBs vaccinated groups compared with the adjuvant-vaccinated, infected control groups. The adjuvant-vaccinated groups had significantly lower IgG1 to IgG2a ratios and lower IL-4 to IFNγ ratios than the FhSAP2-IBs vaccinated animals, which is indicative of higher levels of Th2 immune responses. Irrespective to the adjuvant used, animals vaccinated with FhSAP2-IBs exhibited significantly higher survival percentage and less liver damage than the adjuvant-control groups. This study suggests that FhSAP2 has potential as vaccine against F. hepatica and that the protection elicited by this molecule could be linked to a mechanism driven by the CD4-Th1 cells. Published by Elsevier Inc.

  6. Body temperature changes during simulated bacterial infection in a songbird: fever at night and hypothermia during the day.

    Science.gov (United States)

    Sköld-Chiriac, Sandra; Nord, Andreas; Tobler, Michael; Nilsson, Jan-Åke; Hasselquist, Dennis

    2015-09-01

    Although fever (a closely regulated increase in body temperature in response to infection) typically is beneficial, it is energetically costly and may induce detrimentally high body temperatures. This can increase the susceptibility to energetic bottlenecks and risks of overheating in some organisms. Accordingly, it could be particularly interesting to study fever in small birds, which have comparatively high metabolic rates and high, variable body temperatures. We therefore investigated two aspects of fever and other sickness behaviours (circadian variation, dose dependence) in a small songbird, the zebra finch. We injected lipopolysaccharide (LPS) at the beginning of either the day or the night, and subsequently monitored body temperature, body mass change and food intake for the duration of the response. We found pronounced circadian variation in the body temperature response to LPS injection, manifested by (dose-dependent) hypothermia during the day but fever at night. This resulted in body temperature during the peak response being relatively similar during the day and night. Day-to-night differences might be explained in the context of circadian variation in body temperature: songbirds have a high daytime body temperature that is augmented by substantial heat production peaks during activity. This might require a trade-off between the benefit of fever and the risk of overheating. In contrast, at night, when body temperature is typically lower and less variable, fever can be used to mitigate infection. We suggest that the change in body temperature during infection in small songbirds is context dependent and regulated to promote survival according to individual demands at the time of infection. © 2015. Published by The Company of Biologists Ltd.

  7. Refolding in high hydrostatic pressure of recombinant proteins from inclusion bodies in Escherichia Coli; Renaturacao em altas pressoes hidrostaticas de proteinas recombinantes agregadas em corpos de inclusao produzidos em Escherichia Coli

    Energy Technology Data Exchange (ETDEWEB)

    Balduino, Keli Nunes

    2009-07-01

    The expression of proteins as inclusion bodies in bacteria is a widely used alternative for production of recombinant protein. However, the aggregation is a problem often encountered during refolding of these proteins. High hydrostatic pressure are able to solubilise the inclusion bodies in the presence of low concentrations of denaturant reagents, encouraging refolding protein with high efficiency and reduce costs. This work aims to refolding of recombinant proteins expressed in Escherichia coli from inclusion bodies using high hydrostatic pressure. Three toxins, all featuring five or more disulfide bonds were studied: NXH8, Natterin 2 and Bothropstoxin 1. Suspensions of inclusion bodies of the three proteins were pressurized to 2000 bars for 16 hours. The buffers were optimized for refolding of the three proteins. The buffer used in the refolding of NXH8 was 50 mM Tris HCl, pH 9.0 with proportion of 1GSH: 4GSSG at a concentration of 6 mM and 2 M GdnHCl. Inclusion bodies were used in O.D. (A600nm) of 0.5. After refolding process, dialysis was performed at pH 7.0. The final yield of obtaining soluble NXH8 was 40% (28,6 mg of soluble NXH8/L of culture medium). The refolding of Bothropstoxin 1 was obtained in refolding buffer of Tris HCl 50 mM, pH 7,5 with proportion of 2 GSH: GSSG 3 and concentration of 3 mM and 1 M GdnHCl. Use with a suspension of O.D. (A600nm) of 0.5. The final yield of recovery of Bothropstoxin 1 refolded was 32% (9,2 mg of refolded Bothropstoxin 1/L of culture medium). The refolding of Natterin 2 was performed in the refolding buffer: 20 mM Tris HCl pH 9.0 at a ratio of 2 GSH: 3GSSG and concentration of 10 mM and 1 M GdnHCl and inclusion bodies O.D. (A600nm) of 6.0. The yield of Natterin 2 refolded was 20% (3,7 mg/L of culture medium). Physico-chemical and biological analysis were performed by SDS-PAGE, western blot, scanning electron microscopy, biological tests in vivo and in vitro and structural. The analysis conducted in NXH8 did not show

  8. More than 9,000,000 unique genes in human gut bacterial community: estimating gene numbers inside a human body.

    Science.gov (United States)

    Yang, Xing; Xie, Lu; Li, Yixue; Wei, Chaochun

    2009-06-29

    Estimating the number of genes in human genome has been long an important problem in computational biology. With the new conception of considering human as a super-organism, it is also interesting to estimate the number of genes in this human super-organism. We presented our estimation of gene numbers in the human gut bacterial community, the largest microbial community inside the human super-organism. We got 552,700 unique genes from 202 complete human gut bacteria genomes. Then, a novel gene counting model was built to check the total number of genes by combining culture-independent sequence data and those complete genomes. 16S rRNAs were used to construct a three-level tree and different counting methods were introduced for the three levels: strain-to-species, species-to-genus, and genus-and-up. The model estimates that the total number of genes is about 9,000,000 after those with identity percentage of 97% or up were merged. By combining completed genomes currently available and culture-independent sequencing data, we built a model to estimate the number of genes in human gut bacterial community. The total number of genes is estimated to be about 9 million. Although this number is huge, we believe it is underestimated. This is an initial step to tackle this gene counting problem for the human super-organism. It will still be an open problem in the near future. The list of genomes used in this paper can be found in the supplementary table.

  9. Study of Meteoritic Inclusion

    DEFF Research Database (Denmark)

    Olsen, Mia Bjørg Stolberg

    ionization mass spectrometry (SIMS). Moreover, we have shown that combining careful petrological investigations with high-precision isotope measurements of multiple systems on single meteoritic inclusions can potentially provide unique insights into the formation history of the solar system's earliest solids...... and the observation of a reduced initial abundance of 26Al in the accretion regions of chondrules and asteroidal bodies impacts our understanding of the accretion timescales of protoplanets in a significant way. Combining high-precision isotope measurements of multiple systems on individual meteoritic inclusions...

  10. Comparison and evaluation of experimental mediastinitis models: precolonized foreign body implants and bacterial suspension inoculation seems promising

    Directory of Open Access Journals (Sweden)

    Kose Necmi

    2006-04-01

    Full Text Available Abstract Background Post-sternotomy mediastinitis (PSM is a devastating surgical complication affecting 1–3% of patients that undergo cardiac surgery. Staphylococcus aureus is one of the most commonly encountered bacterial pathogen cultured from mediastinal samples obtained from patients with PSM. A component of the membrane of the gram positive bacteria, lipoteichoic acid, stimulates the blood monocytes and macrophages to secrete cytokines, radicals and nitrogen species leading to oxido-inflammatory damage. This seems to be responsible for the high mortality rate in PSM. For the evaluation of the pathogenesis of infection or for the investigation of alternative treatment models in infection, no standard model of mediastinitis seems to be available. In this study, we evaluated four mediastinitis models in rats. Methods The rats were divided into four groups to form different infection models. Group A: A suspension of 1 × 107 colony-forming units Staphylococcus aureus in 0,5 mL was inoculated from the right second intercostal space into the mediastinum. Group B: A hole was created in the right second intercostal space and a piece of stainless-steel implant with a length of 0.5 cm was inserted into the mediastinum and a suspension of 1 × 107 cfu bacteria in 0,5 mL was administered via the tail vein. Group C: Precolonized stainless-steel implant was inserted into the mediastinum. Group D: Precolonized stainless-steel implant was inserted into the mediastinum and the bacteria suspension was also injected into the mediastinum. On the 10th day, rats were sacrificed and the extension of infection in the mediastenae was evaluated by quantitative cultures. Myeloperoxidase activity (MPO and malondialdehyde (MDA levels were determined in the sera to evaluate the neutrophil activation and assess the inflammatory oxidation. Results The degree of infection in group C and D were 83.3% and 100% respectively (P P Conclusion Infected implants and high

  11. Translating Inclusion

    DEFF Research Database (Denmark)

    Fallov, Mia Arp; Birk, Rasmus

    2018-01-01

    The purpose of this paper is to explore how practices of translation shape particular paths of inclusion for people living in marginalized residential areas in Denmark. Inclusion, we argue, is not an end-state, but rather something which must be constantly performed. Active citizenship, today......, is not merely a question of participation, but of learning to become active in all spheres of life. The paper draws on empirical examples from a multi-sited field work in 6 different sites of local community work in Denmark, to demonstrate how different dimensions of translation are involved in shaping active...... citizenship. We propose the following different dimensions of translation: translating authority, translating language, translating social problems. The paper takes its theoretical point of departure from assemblage urbanism, arguing that cities are heterogeneous assemblages of socio-material interactions...

  12. Study of Meteoritic Inclusion

    DEFF Research Database (Denmark)

    Olsen, Mia Bjørg Stolberg

    . The manuscripts presented in this thesis have provided critical insights into the origin and distribution of short-lived radioisotopes as well as the formation and transport history of chondrules and, by extension, the precursor material to asteroidal and planetary bodies. The proposal of 26Al heterogeneity...... and the observation of a reduced initial abundance of 26Al in the accretion regions of chondrules and asteroidal bodies impacts our understanding of the accretion timescales of protoplanets in a significant way. Combining high-precision isotope measurements of multiple systems on individual meteoritic inclusions...

  13. Social imaginaries and inclusion

    DEFF Research Database (Denmark)

    Hansen, Janne Hedegaard

    2016-01-01

    , the development of inclusive schools and inclusive learning environments will involve both inclusion and exclusion processes. With this starting point, international educational research knowledge about inclusive schools and inclusive learning environments in general will be related to the fundamental dilemma...

  14. Bacterial Keratitis

    Science.gov (United States)

    ... Español Eye Health / Eye Health A-Z Bacterial Keratitis Sections What Is Bacterial Keratitis? Bacterial Keratitis Symptoms ... Lens Care Bacterial Keratitis Treatment What Is Bacterial Keratitis? Leer en Español: ¿Qué Es la Queratitis Bacteriana? ...

  15. Effects of Dietary Inclusion of Oil Sources With or Without Vitamin E on Body Composition and Meat Oxidation Level in Broilers*

    Directory of Open Access Journals (Sweden)

    EÖ Taşdelen

    Full Text Available ABSTRACT The objective of this study was to investigate the effects of rapeseed oil substitution and vitamin E supplemantation on performance and meat quality. 4 replacement levels of rapeseed oil with sunflower oil ( 0, 33.3 , 66.7 and 100% and 2 levels of vitamin E (0 (50 I.U. from vitamin premix 300 mg/kg in a 2 x 3 factorial arrangement for a total of 8 treatments with 4 replicates each, containing 10 day-old male Ross 308 broilers, were examined in the experiment. Dietary treatments had no significant effect on carcass parameters, feed conversion (FCR and mortality (p>0.05. However, a significant interaction was observed between oil replacement and vitamin E: the negative effect of 100% sunflower oil on BWG (p<0.01 was alleviated by the addition of vitamin E. On the other hand, inclusion of rapeseed oil improved BWG (p<0.01. Rapeseed oil substitution significantly increased amount of total n-3 PUFA and decreased n-6:n-3 ratio in thigh and breast meat (p<0.01. Vitamin E supplemantation contributed to deposition of n-3 PUFA (p<0.01. Replacement of rapeseed oil without vitamin E tended to increase (p<0.01 malonaldehyde production. However, the dietary supplementation of vitamin E markedly (p<0.01 decreased the susceptibility of meat to peroxidation. Inclusion of rapeseed oil did not cause any negative perception on olfactory, texture, and taste of broiler meat. So, it can be concluded that rapeseed oil substitution significantly increased n-3 PUFA deposition without altering performance and sensory properties of broiler meat and, vitamin E had strong potential to prevent the meat lipids from oxidation.

  16. Detergent Insoluble Proteins and Inclusion Body-Like Structures Immunoreactive for PRKDC/DNA-PK/DNA-PKcs, FTL, NNT, and AIFM1 in the Amygdala of Cognitively Impaired Elderly Persons.

    Science.gov (United States)

    Gal, Jozsef; Chen, Jing; Katsumata, Yuriko; Fardo, David W; Wang, Wang-Xia; Artiushin, Sergey; Price, Douglas; Anderson, Sonya; Patel, Ela; Zhu, Haining; Nelson, Peter T

    2018-01-01

    Misfolded protein in the amygdala is a neuropathologic feature of Alzheimer disease and many other neurodegenerative disorders. We examined extracts from human amygdala (snap-frozen at autopsy) to investigate whether novel and as yet uncharacterized misfolded proteins would be detectable. Polypeptides from the detergent-insoluble, urea-soluble protein fractions of amygdala were interrogated using liquid chromatography-electrospray ionization-tandem mass spectrometry. Among the detergent-insoluble proteins identified in amygdala of demented subjects but not controls were Tau, TDP-43, Aβ, α-synuclein, and ApoE. Additional detergent-insoluble proteins from demented subjects in the high-molecular weight portion of SDS gels included NNT, TNIK, PRKDC (DNA-PK, or DNA-PKcs), ferritin light chain (FTL), AIFM1, SYT11, STX1B, EAA1, COL25A1, M4K4, CLH1, SQSTM, SYNJ1, C3, and C4. In follow-up immunohistochemical experiments, NNT, TNIK, PRKDC, AIFM1, and FTL were observed in inclusion body-like structures in cognitively impaired subjects' amygdalae. Double-label immunofluorescence revealed that FTL and phospho-PRKDC immunoreactivity colocalized partially with TDP-43 and/or Tau inclusion bodies. Western blots showed high-molecular weight "smears", particularly for NNT and PRKDC. A preliminary genetic association study indicated that rare NNT, TNIK, and PRKDC gene variants had nominally significant association with Alzheimer-type dementia risk. In summary, novel detergent-insoluble proteins, with evidence of proteinaceous deposits, were found in amygdalae of elderly, cognitively impaired subjects. © 2017 American Association of Neuropathologists, Inc. All rights reserved.

  17. Bacterial meningitis

    NARCIS (Netherlands)

    Roos, Karen L.; van de Beek, Diederik

    2010-01-01

    Bacterial meningitis is a neurological emergency. Empiric antimicrobial and adjunctive therapy should be initiated as soon as a single set of blood cultures has been obtained. Clinical signs suggestive of bacterial meningitis include fever, headache, meningismus, vomiting, photophobia, and an

  18. Singing and social inclusion.

    Science.gov (United States)

    Welch, Graham F; Himonides, Evangelos; Saunders, Jo; Papageorgi, Ioulia; Sarazin, Marc

    2014-01-01

    There is a growing body of neurological, cognitive, and social psychological research to suggest the possibility of positive transfer effects from structured musical engagement. In particular, there is evidence to suggest that engagement in musical activities may impact on social inclusion (sense of self and of being socially integrated). Tackling social exclusion and promoting social inclusion are common concerns internationally, such as in the UK and the EC, and there are many diverse Government ministries and agencies globally that see the arts in general and music in particular as a key means by which social needs can be addressed. As part of a wider evaluation of a national, Government-sponsored music education initiative for Primary-aged children in England ("Sing Up"), opportunity was taken by the authors, at the request of the funders, to assess any possible relationship between (a) children's developing singing behavior and development and (b) their social inclusion (sense of self and of being socially integrated). Subsequently, it was possible to match data from n = 6087 participants, drawn from the final 3 years of data collection (2008-2011), in terms of each child's individually assessed singing ability (based on their singing behavior of two well-known songs to create a "normalized singing score") and their written responses to a specially-designed questionnaire that included a set of statements related to children's sense of being socially included to which the children indicated their level of agreement on a seven-point Likert scale. Data analyses suggested that the higher the normalized singing development rating, the more positive the child's self-concept and sense of being socially included, irrespective of singer age, sex and ethnicity.

  19. Singing and social inclusion

    Science.gov (United States)

    Welch, Graham F.; Himonides, Evangelos; Saunders, Jo; Papageorgi, Ioulia; Sarazin, Marc

    2014-01-01

    There is a growing body of neurological, cognitive, and social psychological research to suggest the possibility of positive transfer effects from structured musical engagement. In particular, there is evidence to suggest that engagement in musical activities may impact on social inclusion (sense of self and of being socially integrated). Tackling social exclusion and promoting social inclusion are common concerns internationally, such as in the UK and the EC, and there are many diverse Government ministries and agencies globally that see the arts in general and music in particular as a key means by which social needs can be addressed. As part of a wider evaluation of a national, Government-sponsored music education initiative for Primary-aged children in England (“Sing Up”), opportunity was taken by the authors, at the request of the funders, to assess any possible relationship between (a) children's developing singing behavior and development and (b) their social inclusion (sense of self and of being socially integrated). Subsequently, it was possible to match data from n = 6087 participants, drawn from the final 3 years of data collection (2008–2011), in terms of each child's individually assessed singing ability (based on their singing behavior of two well-known songs to create a “normalized singing score”) and their written responses to a specially-designed questionnaire that included a set of statements related to children's sense of being socially included to which the children indicated their level of agreement on a seven-point Likert scale. Data analyses suggested that the higher the normalized singing development rating, the more positive the child's self-concept and sense of being socially included, irrespective of singer age, sex and ethnicity. PMID:25120514

  20. Singing and social inclusion

    Directory of Open Access Journals (Sweden)

    Graham Frederick Welch

    2014-07-01

    Full Text Available There is a growing body of neurological, cognitive and social psychological research to suggest the possibility of positive transfer effects from structured musical engagement. In particular, there is evidence to suggest that engagement in musical activities may impact on social inclusion (sense of self and of being socially integrated. Tackling social exclusion and promoting social inclusion are common concerns internationally, such as in the UK and the EC, and there are many diverse Government ministries and agencies globally that see the arts in general and music in particular as a key means by which social needs can be addressed. As part of a wider evaluation of a national, Government-sponsored music education initiative for Primary-aged children in England (‘Sing Up’, opportunity was taken by the authors, at the request of the funders, to assess any possible relationship between (a children’s developing singing behaviour and development and (b their social inclusion (sense of self and of being socially integrated. Subsequently, it was possible to match data from n=6087 participants, drawn from the final three years of data collection (2008-2011, in terms of each child’s individually assessed singing ability (based on their singing behaviour of two well-known songs to create a 'normalised singing score' and their written responses to a specially-designed questionnaire that included a set of statements related to children’s sense of being socially included to which the children indicated their level of agreement on a seven-point Likert scale. Data analyses suggested that the higher the normalized singing development rating, the more positive the child’s self-concept and sense of being socially included, irrespective of singer age, sex and ethnicity.

  1. Limits to Inclusion

    Science.gov (United States)

    Hansen, Janne Hedegaard

    2012-01-01

    In this article, I will argue that a theoretical identification of the limit to inclusion is needed in the conceptual identification of inclusion. On the one hand, inclusion is formulated as a vision that is, in principle, limitless. On the other hand, there seems to be an agreement that inclusion has a limit in the pedagogical practice. However,…

  2. Genetics Home Reference: inclusion body myopathy 2

    Science.gov (United States)

    ... molecules, sialic acid influences a wide variety of cellular functions including cell movement (migration), attaching cells to one another (adhesion), signaling between cells, and inflammation. The mutations responsible ...

  3. Bacterial Proteasomes.

    Science.gov (United States)

    Jastrab, Jordan B; Darwin, K Heran

    2015-01-01

    Interest in bacterial proteasomes was sparked by the discovery that proteasomal degradation is required for the pathogenesis of Mycobacterium tuberculosis, one of the world's deadliest pathogens. Although bacterial proteasomes are structurally similar to their eukaryotic and archaeal homologs, there are key differences in their mechanisms of assembly, activation, and substrate targeting for degradation. In this article, we compare and contrast bacterial proteasomes with their archaeal and eukaryotic counterparts, and we discuss recent advances in our understanding of how bacterial proteasomes function to influence microbial physiology.

  4. Bacterial adhesion

    NARCIS (Netherlands)

    Loosdrecht, van M.C.M.

    1988-01-01

    As mentioned in the introduction of this thesis bacterial adhesion has been studied from a variety of (mostly practice oriented) starting points. This has resulted in a range of widely divergent approaches. In order to elucidate general principles in bacterial adhesion phenomena, we felt it

  5. Roadmap for Inclusive Innovation

    Data.gov (United States)

    Allegheny County / City of Pittsburgh / Western PA Regional Data Center — The Roadmap for Inclusive Innovation data set shares the status of action items under the Roadmap for Inclusive Innovation. Each action or project has been assigned...

  6. Teachers becoming inclusive practitioners

    African Journals Online (AJOL)

    , construct their identities in the light of inclusive education, and how they negotiate the tensions and contradictions emerging from the processof becoming inclusive practitioners. Central to this discussion is the understanding that teachers' ...

  7. Foresighting for Inclusive Development

    DEFF Research Database (Denmark)

    Andersen, Allan Dahl; Andersen, Per Dannemand

    2017-01-01

    We propose that foresight can contribute to inclusive development by making innovation systems more inclusive. Processes of developing future oriented innovation policies are often unsuccessful and rarely inclusive. We conceptualize such processes as foresighting. We focus on how the ex-ante desi...... in foresight cases in two emerging economies: Brazil and South Korea. We conclude that better systemic and innovation oriented foresight is needed to enhance inclusive development....

  8. Inclusion: Conceptualization and measurement

    NARCIS (Netherlands)

    Jansen, W.S.; Otten, S.; Van der Zee, K.I.; Jans, L.

    2014-01-01

    n the present research, we introduced a conceptual framework of inclusion and subsequently used this as a starting point to develop and validate a scale to measure perceptions of inclusion. Departing from existing work on inclusion and complementing this with theoretical insights from optimal

  9. Inclusion: Conceptualization and measurement

    NARCIS (Netherlands)

    Jansens, Wiebren S.; Otten, Sabine; van der Zee, Karen; Jans, Lise

    2014-01-01

    In the present research, we introduced a conceptual framework of inclusion and subsequently used this as a starting point to develop and validate a scale to measure perceptions of inclusion. Departing from existing work on inclusion and complementing this with theoretical insights from optimal

  10. Inclusion : Conceptualization and measurement

    NARCIS (Netherlands)

    Jansen, Wiebren S.; Otten, Sabine; van der Zee, Karen I.; Jans, Lise

    In the present research, we introduced a conceptual framework of inclusion and subsequently used this as a starting point to develop and validate a scale to measure perceptions of inclusion. Departing from existing work on inclusion and complementing this with theoretical insights from optimal

  11. Inclusion : Conceptualization and measurement

    NARCIS (Netherlands)

    Jansen, Wiebren S.; Otten, Sabine; van der Zee, Karen I.; Jans, Lise

    2014-01-01

    In the present research, we introduced a conceptual framework of inclusion and subsequently used this as a starting point to develop and validate a scale to measure perceptions of inclusion. Departing from existing work on inclusion and complementing this with theoretical insights from optimal

  12. The Full Inclusion Movement.

    Science.gov (United States)

    Machado, Rodney E.; And Others

    1996-01-01

    Overviews background of the movement toward full inclusion of special education students into regular classrooms, including legal issues and successful educational practices. Suggests that full inclusion does not benefit all students and that inclusion should be one of several alternatives to meeting students' educational needs. Of approximately…

  13. Footstep towards Inclusive Education

    Science.gov (United States)

    Abbas, Faiza; Zafar, Aneeka; Naz, Tayyaba

    2016-01-01

    Inclusive education is a rising trend in the world. The first step towards inclusive education is providing the awareness to the general education teachers. This study focused to investigate the general education teachers of primary and secondary level awareness about the special education and inclusive education. This study is descriptive method…

  14. Documenting Your Inclusion Efforts.

    Science.gov (United States)

    Fritz, Marie F.; Miller, Maury

    This paper briefly addresses critical issues in documenting studies of the effectiveness of inclusive education for students with disabilities and includes a summary table of the existing literature and guidelines for problem solving about inclusion. The importance of sound methodology in documenting effects of inclusion and the dangers of value…

  15. Analysis for the presence of determinants involved in the transport of mercury across bacterial membrane from polluted water bodies of India.

    Science.gov (United States)

    Jan, Arif Tasleem; Azam, Mudsser; Choi, Inho; Ali, Arif; Haq, Qazi Mohd Rizwanul

    2016-01-01

    Mercury, which is ubiquitous and recalcitrant to biodegradation processes, threatens human health by escaping to the environment via various natural and anthropogenic activities. Non-biodegradability of mercury pollutants has necessitated the development and implementation of economic alternatives with promising potential to remove metals from the environment. Enhancement of microbial based remediation strategies through genetic engineering approaches provides one such alternative with a promising future. In this study, bacterial isolates inhabiting polluted sites were screened for tolerance to varying concentrations of mercuric chloride. Following identification, several Pseudomonas and Klebsiella species were found to exhibit the highest tolerance to both organic and inorganic mercury. Screened bacterial isolates were examined for their genetic make-up in terms of the presence of genes (merP and merT) involved in the transport of mercury across the membrane either alone or in combination to deal with the toxic mercury. Gene sequence analysis revealed that the merP gene showed 86-99% homology, while the merT gene showed >98% homology with previously reported sequences. By exploring the genes involved in imparting metal resistance to bacteria, this study will serve to highlight the credentials that are particularly advantageous for their practical application to remediation of mercury from the environment. Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  16. Foresighting for Inclusive Development

    DEFF Research Database (Denmark)

    Dahl Andersen, Allan; Andersen, Per Dannemand

    We propose that foresight can contribute to inclusive development by making innovation policy processes more inclusive, which in turn makes innovation systems more inclusive. Processes of developing future-oriented innovation policies are often unsuccessful and rarely inclusive. We conceptualize...... and transform innovation systems because a set of distributed actors, rather than ministries and innovation agencies, is the gatekeepers of change. From this perspective, inclusion is a precondition rather than an obstacle for transformation. We develop a conceptual framework and use it to study design...

  17. Bacterial Vaginosis

    Science.gov (United States)

    ... Archive STDs Home Page Bacterial Vaginosis (BV) Chlamydia Gonorrhea Genital Herpes Hepatitis HIV/AIDS & STDs Human Papillomavirus ( ... of getting other STDs, such as chlamydia and gonorrhea . These bacteria can sometimes cause pelvic inflammatory disease ( ...

  18. Deploying and implementing Inclusive Physical Education

    Directory of Open Access Journals (Sweden)

    Eliana Lúcia Ferreira

    2014-04-01

    Full Text Available Physical Education, as a curricular component of basic education, is not indifferent to the movement of Inclusive physical education. Differentiated bodies are conquering new social spaces. Our aim through this investigation is to identify the main historical practices regarding Brazilian Policy of Inclusive Education and to point out proposals to implement inclusive Physical education. Our methodology consists of a descriptive study based on two main axes. The first axis is related to a historical discussion whose source was national documents of reference about Inclusive Physical Education. The second axis is related to the experience of 2000 teachers; we have the occasion to analyze their practices on the subject of Inclusive physical education. As a result, this investigation also develops a proposal to inclusive physical education; this proposal is established in another dimension and understanding of work and movement. Concerning the final considerations we suggest a disruption with already crystallized bodily practices and we suggest actions respecting individual differences. Moreover, we point out the fact that Inclusive Physical Education has advocated another meaning for the body, in favor of a more collective physical education and searching for activities in which individuality prevails.

  19. INCLUSIVE EDUCATION IN ITALY

    OpenAIRE

    Voitkāne, Vita

    2017-01-01

    European member States implement Inclusive Education policies thus contributing to a sustainable, inclusive society, although each country is at a different stage in this process. Italy, one of the first countries to launch integrative learning, has set an example since the 1970s, although the quality of inclusive education is unpredictable due to many issues. Authors Cantoni and Panetta (2006) emphasize that, although the culture of integration in Italy exists, much needs to be done to impro...

  20. SUPPORT IN INCLUSIVE EDUCATION

    OpenAIRE

    Belma Čičkušić; Ševala Tulumović; Selma Bakić; Salem Bakić

    2016-01-01

    In order for inclusive class to be successful, associates are of great help to teachers. Besides associates, teachers' specialization can be accomplished through educational seminars on the inclusion topic. However, information about inclusion, working with children with special needs, can also be found in scientific journals that offer more information on methods of working with children with special needs, didactic materials customized according to abilities of children. Aim of ...

  1. Confronting New Demands : Inclusive Growth, Inclusive Trade ...

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

    The research agenda will cover four themes: linkages between trade and inclusive growth; negotiation issues with poorly understood implications, such as labour standards and intellectual property; liberalization of trade in services, and the emerging regional trade policy architecture. The idea is to promote a better ...

  2. BACTERIAL CONSORTIUM

    Directory of Open Access Journals (Sweden)

    Payel Sarkar

    2013-01-01

    Full Text Available Petroleum aromatic hydrocarbons like benzen e, toluene, ethyl benzene and xylene, together known as BTEX, has almost the same chemical structure. These aromatic hydrocarbons are released as pollutants in th e environment. This work was taken up to develop a solvent tolerant bacterial cons ortium that could degrade BTEX compounds as they all share a common chemical structure. We have isolated almost 60 different types of bacterial strains from different petroleum contaminated sites. Of these 60 bacterial strains almost 20 microorganisms were screene d on the basis of capability to tolerate high concentration of BTEX. Ten differe nt consortia were prepared and the compatibility of the bacterial strains within the consortia was checked by gram staining and BTEX tolerance level. Four successful mi crobial consortia were selected in which all the bacterial strains concomitantly grew in presence of high concentration of BTEX (10% of toluene, 10% of benzene 5% ethyl benzene and 1% xylene. Consortium #2 showed the highest growth rate in pr esence of BTEX. Degradation of BTEX by consortium #2 was monitored for 5 days by gradual decrease in the volume of the solvents. The maximum reduction observed wa s 85% in 5 days. Gas chromatography results also reveal that could completely degrade benzene and ethyl benzene within 48 hours. Almost 90% degradation of toluene and xylene in 48 hours was exhibited by consortium #2. It could also tolerate and degrade many industrial solvents such as chloroform, DMSO, acetonitrile having a wide range of log P values (0.03–3.1. Degradation of aromatic hydrocarbon like BTEX by a solvent tolerant bacterial consortium is greatly significant as it could degrade high concentration of pollutants compared to a bacterium and also reduces the time span of degradation.

  3. Understanding Inclusion in Cyprus

    Science.gov (United States)

    Mamas, Christoforos

    2013-01-01

    This paper provides a framework for understanding inclusion in Cyprus. The evidence base is the result of a six-month qualitative research study in five Cypriot mainstream primary schools. Despite the rhetoric in favour of inclusion, it seems that the Cypriot educational system is still highly segregating in its philosophy and does not fully…

  4. School Inclusion Programmes (SIPS)

    Science.gov (United States)

    Drossinou-Korea, Maria; Matousi, Dimitra; Panopoulos, Nikolaos; Paraskevopoulou, Aikaterini

    2016-01-01

    The purpose of this work was to understand the school inclusion programmes (SIPs) for students with special educational needs (SEN). The methodology was conducted in the field of special education (SE) and focuses on three case studies of students who was supported by SIPs. The Targeted, Individual, Structured, Inclusion Programme for students…

  5. Ladle Metallurgy Kinetics: Inclusion-Inclusion Reactions

    Science.gov (United States)

    Pistorius, P. Chris

    An example is presented to illustrate the joint effect of local reaction equilibria and mass transfer limitations, for reactions during ladle refining of steel. The example relies on some of the kinetic principles that David Robertson has employed to quantify many metallurgical processes. In calcium treatment of alumina inclusions in aluminum-killed steels, solid CaS forms as an intermediate reaction product. During subsequent reaction, CaS disappears and calcium aluminate forms; at the same time, aluminum and sulfur dissolve in the steel. Kinetic analysis shows that the rate of this reaction is not limited by mass transfer of dissolved aluminum and sulfur away from the reacting inclusions. The reaction rate is likely limited by transport of dissolved calcium. This example also illustrates the use of FactSage macros for kinetic modeling.

  6. Bacterial Ecology

    DEFF Research Database (Denmark)

    Fenchel, Tom

    2011-01-01

    Bacterial ecology is concerned with the interactions between bacteria and their biological and nonbiological environments and with the role of bacteria in biogeochemical element cycling. Many fundamental properties of bacteria are consequences of their small size. Thus, they can efficiently exploit...

  7. Bacterial meningitis

    NARCIS (Netherlands)

    Heckenberg, Sebastiaan G. B.; Brouwer, Matthijs C.; van de Beek, Diederik

    2014-01-01

    Bacterial meningitis is a neurologic emergency. Vaccination against common pathogens has decreased the burden of disease. Early diagnosis and rapid initiation of empiric antimicrobial and adjunctive therapy are vital. Therapy should be initiated as soon as blood cultures have been obtained,

  8. Bacterial lipases

    NARCIS (Netherlands)

    Jaeger, Karl-Erich; Ransac, Stéphane; Dijkstra, Bauke W.; Colson, Charles; Heuvel, Margreet van; Misset, Onno

    Many different bacterial species produce lipases which hydrolyze esters of glycerol with preferably long-chain fatty acids. They act at the interface generated by a hydrophobic lipid substrate in a hydrophilic aqueous medium. A characteristic property of lipases is called interfacial activation,

  9. Bacterial Ecology

    DEFF Research Database (Denmark)

    Fenchel, Tom

    2011-01-01

    , the production and oxidation of methane, nitrate reduction and fixation of atmospheric nitrogen are exclusively carried out by different groups of bacteria. Some bacterial species – ‘extremophiles’ – thrive in extreme environments in which no eukaryotic organisms can survive with respect to temperature, salinity...

  10. Bacterial Vaginosis

    Science.gov (United States)

    ... that coats the walls of the vagina Vaginal discharge with an unpleasant or fishlike odor Vaginal pain or itching Burning during urination Doctors are unsure of the incubation period for bacterial vaginosis. How Is the Diagnosis Made? Your child’s pediatrician can make the diagnosis ...

  11. Bacterial stress

    Indian Academy of Sciences (India)

    First page Back Continue Last page Graphics. Bacterial stress. Physicochemical and chemical parameters: temperature, pressure, pH, salt concentration, oxygen, irradiation. Nutritional depravation: nutrient starvation, water shortage. Toxic compounds: Antibiotics, heavy metals, toxins, mutagens. Interactions with other cells: ...

  12. Bacterial Adhesion & Blocking Bacterial Adhesion

    DEFF Research Database (Denmark)

    Vejborg, Rebecca Munk

    2008-01-01

    reduce or delay bacterial biofilm formation of a range of urinary tract infectious E.coli and Klebsiella isolates. Several other proteinaceous coatings were also found to display anti-adhesive properties, possibly providing a measure for controlling the colonization of implant materials. Several other...... components. These substances may both mediate and stabilize the bacterial biofilm. Finally, several adhesive structures were examined, and a novel physiological biofilm phenotype in E.coli biofilms was characterized, namely cell chain formation. The autotransporter protein, antigen 43, was implicated...

  13. [Bacterial biofilms and infection].

    Science.gov (United States)

    Lasa, I; Del Pozo, J L; Penadés, J R; Leiva, J

    2005-01-01

    In developed countries we tend to think of heart disease and the numerous forms of cancer as the main causes of mortality, but on a global scale infectious diseases come close, or may even be ahead: 14.9 million deaths in 2002 compared to cardiovascular diseases (16.9 million deaths) and cancer (7.1 million deaths) (WHO report 2004). The infectious agents responsible for human mortality have evolved as medical techniques and hygienic measures have changed. Modern-day acute infectious diseases caused by specialized bacterial pathogens such as diphtheria, tetanus, cholera, plague, which represented the main causes of death at the beginning of XX century, have been effectively controlled with antibiotics and vaccines. In their place, more than half of the infectious diseases that affect mildly immunocompromised patients involve bacterial species that are commensal with the human body; these can produce chronic infections, are resistant to antimicrobial agents and there is no effective vaccine against them. Examples of these infections are the otitis media, native valve endocarditis, chronic urinary infections, bacterial prostatitis, osteomyelitis and all the infections related to medical devices. Direct analysis of the surface of medical devices or of tissues that have been foci of chronic infections shows the presence of large numbers of bacteria surrounded by an exopolysaccharide matrix, which has been named the "biofilm". Inside the biofilm, bacteria grow protected from the action of the antibodies, phagocytic cells and antimicrobial treatments. In this article, we describe the role of bacterial biofilms in human persistent infections.

  14. Fluid inclusion geothermometry

    Science.gov (United States)

    Cunningham, C.G.

    1977-01-01

    Fluid inclusions trapped within crystals either during growth or at a later time provide many clues to the histories of rocks and ores. Estimates of fluid-inclusion homogenization temperature and density can be obtained using a petrographic microscope with thin sections, and they can be refined using heating and freezing stages. Fluid inclusion studies, used in conjunction with paragenetic studies, can provide direct data on the time and space variations of parameters such as temperature, pressure, density, and composition of fluids in geologic environments. Changes in these parameters directly affect the fugacity, composition, and pH of fluids, thus directly influencing localization of ore metals. ?? 1977 Ferdinand Enke Verlag Stuttgart.

  15. Microvillous inclusion disease (microvillous atrophy

    Directory of Open Access Journals (Sweden)

    Goulet Olivier

    2006-06-01

    Full Text Available Abstract Microvillous inclusion disease (MVID or microvillous atrophy is a congenital disorder of the intestinal epithelial cells that presents with persistent life-threatening watery diarrhea and is characterized by morphological enterocyte abnormalities. MVID manifests either in the first days of life (early-onset form or in the first two months (late-onset form of life. MVID is a very rare disorder of unknown origin, probably transmitted as an autosomal recessive trait. To date, no prevalence data are available. Ultrastructural analyses reveal: 1 a partial to total atrophy of microvilli on mature enterocytes with apical accumulation of numerous secretory granules in immature enterocytes; 2 the highly characteristic inclusion bodies containing rudimentary or fully differentiated microvilli in mature enterocytes. Light microscopy shows accumulation of PAS-positive granules at the apical pole of immature enterocytes, together with atrophic band indicating microvillus atrophy and, in parallel, an intracellular PAS or CD10 positive line (marking the microvillous inclusion bodies seen on electron microscopy. Intestinal failure secondary to diarrhea is definitive. To date, no curative therapy exists and children with MVID are totally dependent on parenteral nutrition. Long-term outcome is generally poor, due to metabolic decompensation, repeated states of dehydration, infectious and liver complications related to the parenteral nutrition. As MVID is a very rare disorder, which is extremely difficult to diagnose and manage, children with MVID should be transferred to specialized pediatric gastro-intestinal centers, if possible, a center equipped to perform small bowel transplantation. Early small bowel transplantation resulting in intestinal autonomy gives new hope for disease management and outcome.

  16. Evidence on Inclusion

    DEFF Research Database (Denmark)

    Dyssegaard, Camilla Brørup; Larsen, Michael Søgaard

    The purpose of this publication is to examine existing research on inclusion to identify strategies of inclusion that have generated positive effects. To do so it is necessary to understand the effect of the applied strategies. One approach, which is being discussed, is to use evidence to determine...... which methods have proven more effective than others. The desire to gain insight into research on inclusion forms the basis of the current systematic review. The task was to determine which strategies primary research has found to be most effective for inclusion purposes. We have solved this task...... by addressing the existing research with the following question: What is the effect of including children with special needs in mainstream teaching in basic school, and which of the applied educational methods have proved to have a positive effect?...

  17. Mapping Inclusive Growth

    OpenAIRE

    Raquel Almeida Ramos; Rafael Ranieri; Jan-Willem Lammes

    2013-01-01

    Since its emergence around the turn of the century the concept of inclusive growth has been substantially debated by researchers and practitioners, has increasingly gained space in domestic and international political circles, has inspired the design of and generated projects and policies, and has impacted the views and the lives of many people. While the conceptual debate and the practical application of various definitions of inclusive growth have taken place simultaneously, they have recen...

  18. RISKS OF INCLUSIVE EDUCATION

    OpenAIRE

    M. R. Husnutdinova

    2017-01-01

    Introduction. Inclusion is a new and unfamiliar phenomenon for most of the Russians which is treated as ensuring equal access to education for all students taking into account a variety of their special educational needs and individual capabilities. Inclusive educational model began to take root in Russia without a broad public debate and today’s parents and teachers were not ready to the cardinal changes caused by transition to the new model of education. In this regard, the studying of dire...

  19. Bacterial lipases

    OpenAIRE

    Jaeger, Karl-Erich; Ransac, Stéphane; Dijkstra, Bauke W.; Colson, Charles; Heuvel, Margreet van; Misset, Onno

    1994-01-01

    Many different bacterial species produce lipases which hydrolyze esters of glycerol with preferably long-chain fatty acids. They act at the interface generated by a hydrophobic lipid substrate in a hydrophilic aqueous medium. A characteristic property of lipases is called interfacial activation, meaning a sharp increase in lipase activity observed when the substrate starts to form an emulsion, thereby presenting to the enzyme an interfacial area. As a consequence, the kinetics of a lipase rea...

  20. Limitations of inclusive fitness.

    Science.gov (United States)

    Allen, Benjamin; Nowak, Martin A; Wilson, Edward O

    2013-12-10

    Until recently, inclusive fitness has been widely accepted as a general method to explain the evolution of social behavior. Affirming and expanding earlier criticism, we demonstrate that inclusive fitness is instead a limited concept, which exists only for a small subset of evolutionary processes. Inclusive fitness assumes that personal fitness is the sum of additive components caused by individual actions. This assumption does not hold for the majority of evolutionary processes or scenarios. To sidestep this limitation, inclusive fitness theorists have proposed a method using linear regression. On the basis of this method, it is claimed that inclusive fitness theory (i) predicts the direction of allele frequency changes, (ii) reveals the reasons for these changes, (iii) is as general as natural selection, and (iv) provides a universal design principle for evolution. In this paper we evaluate these claims, and show that all of them are unfounded. If the objective is to analyze whether mutations that modify social behavior are favored or opposed by natural selection, then no aspect of inclusive fitness theory is needed.

  1. RISKS OF INCLUSIVE EDUCATION

    Directory of Open Access Journals (Sweden)

    M. R. Husnutdinova

    2017-01-01

    Full Text Available Introduction. Inclusion is a new and unfamiliar phenomenon for most of the Russians which is treated as ensuring equal access to education for all students taking into account a variety of their special educational needs and individual capabilities. Inclusive educational model began to take root in Russia without a broad public debate and today’s parents and teachers were not ready to the cardinal changes caused by transition to the new model of education. In this regard, the studying of directly educational process by consequences of inclusive training and education is urgent now.The aims of the research are the following: to identify the major risks that characterize the current stage of the implementation process of inclusion in the Russian educational organizations; to consider the main causes that lead to their occurrence; to present a comparative analysis of the views of respondents in comprehensive, inclusive and correctional schools. Methodology and research methods. The Sector Monitoring Studies of Moscow State University of Psychology & Education in 2010 and 2014 implemented a sociological study on the process of introduction of inclusion in the Russian schools. 200 teachers and 244 parents were interviewed in 2010; in 2014, in addition, 178 teachers and 386 parents were interviewed; 47 senior students including those with disabilities were interviewed too.Results. According to the results, the main concerns of the parents of students of comprehensive, inclusive and correctional schools are reduced to a few basic risks: lack of individual approach while teaching children with different educational needs, increased emotional pressure on the child, and child’s perception of the complexity of disability as an equal. These risks arise primarily because of the acute shortage of especially prepared-governmental teachers and socio-cultural, psychological unpreparedness of most contemporary children with disabilities to the perception of

  2. Bacterial mitosis

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Borch, Jonas; Dam, Mette

    2003-01-01

    Bacterial DNA segregation takes place in an active and ordered fashion. In the case of Escherichia coli plasmid R1, the partitioning system (par) separates paired plasmid copies and moves them to opposite cell poles. Here we address the mechanism by which the three components of the R1 par system...... movement is powered by insertional polymerization of ParM. Consistently, we find that segregating plasmids are positioned at the ends of extending ParM filaments. Thus, the process of R1 plasmid segregation in E. coli appears to be mechanistically analogous to the actin-based motility operating...

  3. Bacterial lipases for biotechnological applications

    NARCIS (Netherlands)

    Jaeger, Karl-Erich; Schneidinger, Bernd; Rosenau, Frank; Werner, Michael; Lang, Dietmar; Dijkstra, Bauke W.; Schimossek, Klaus; Zonta, Albin; Reetz, Manfred T.

    1997-01-01

    Lipase genes originating from the Gram-negative bacteria Serrutiu marcescens and Pseudomonus urruginosa were cloned. S. marcescens lipase was overexpressed in Escherichia coli yielding inclusion bodies which were purified and finally refolded to give enzymatically active lipase. The lipase operon of

  4. Nonlinear elastic inclusions in isotropic solids

    KAUST Repository

    Yavari, A.

    2013-10-16

    We introduce a geometric framework to calculate the residual stress fields and deformations of nonlinear solids with inclusions and eigenstrains. Inclusions are regions in a body with different reference configurations from the body itself and can be described by distributed eigenstrains. Geometrically, the eigenstrains define a Riemannian 3-manifold in which the body is stress-free by construction. The problem of residual stress calculation is then reduced to finding a mapping from the Riemannian material manifold to the ambient Euclidean space. Using this construction, we find the residual stress fields of three model systems with spherical and cylindrical symmetries in both incompressible and compressible isotropic elastic solids. In particular, we consider a finite spherical ball with a spherical inclusion with uniform pure dilatational eigenstrain and we show that the stress in the inclusion is uniform and hydrostatic. We also show how singularities in the stress distribution emerge as a consequence of a mismatch between radial and circumferential eigenstrains at the centre of a sphere or the axis of a cylinder.

  5. Creative activity and inclusion

    Directory of Open Access Journals (Sweden)

    Shemanov A.Yu.

    2013-09-01

    Full Text Available The aim of the article was to analyze the inclusion potential of art creative activity, namely of theatre performance, in people with disabilities. The article provides examples of disagreements in understanding the significance of these art activities for exercising the rights of people with disabilities to contribute to culture and art and some problems arising here. The conclusion is made that theatre art performed by people with disabilities is gradually changing its function: from being a means of self-affirmation to the determination of its specific place in overall theatre process. These changes confirm the inclusion potential of theatre art activity.

  6. Lead inclusions in aluminium

    International Nuclear Information System (INIS)

    Johnson, E.; Johansen, A.; Sarholt-Kristensen, L.; Andersen, H.H.; Grabaek, L.; Bohr, J.

    1990-01-01

    Ion implantation at room temperature of lead into aluminum leads to spontaneous phase separation and formation of lead precipitates growing topotactically with the matrix. Unlike the highly pressurized (∼ 1-5 GPa) solid inclusions formed after noble gas implantations, the pressure in the lead precipitates is found to be less than 0.12 GPa. Recently the authors have observed the result that the lead inclusions in aluminum exhibit both superheating and supercooling. In this paper they review and elaborate on these results. Small implantation-induced lead precipitates embedded in an aluminum matrix were studied by x-ray diffraction

  7. Inclusive Education in Israel

    OpenAIRE

    O. V. Mayzel

    2013-01-01

    To get acquainted with the practice of inclusive education in mainstream schools, with professionals who work with special children, to visit the specialist centers to share experiences - all of this was part of an internship program «Early Childhood Education for Children with Special Needs», held in Israel (April 8 -02 May 2013) this year. The country has been selected for an internship, because the practice of inclusive education has been used for over 20 years in Israel. Moreover, a lot ...

  8. Developing a financial inclusion index and inclusive growth in India

    OpenAIRE

    Susanta Kumar SETHY

    2016-01-01

    Financial inclusion is one of the systems through which Inclusive Growth can be achieved in developing countries like India where large sections are unable or hopeless to contribute in the financial system. An inclusive financial system mobilizes more resources for productive purposes leading to higher economic growth, better opportunities and reduction of poverty. This study, proposed an Index of financial inclusion – a multidimensional measure. The Financial Inclusion Index c...

  9. Difference, inclusion, and mathematics education

    DEFF Research Database (Denmark)

    Figueiras, Lourdes; Healy, Lulu; Skovsmose, Ole

    2016-01-01

    The round-table discussion on Difference, Inclusion and Mathematics Education was in included in the scientific programme of VI SIPEM in recognition and celebration of the emerging body of research into the challenges of building a culture of mathematics education which values and respects...... the diversity of learners in different educational contexts – in Brazil and beyond. This paper presents the contributions to the discussion, which focus on the problematisation of the term “inclusion”, explorations of how the practices of previously marginalized students can bring new resources to the teaching...... and learning of mathematics and reflections upon the potentially discriminatory nature of the structures which currently mould school mathematics. The paper aims to serve as material for the developing research agenda of the thirteenth working group of the Brazilian Society of Mathematics Education, which met...

  10. Cytochemical localization of glycogen in Chlamydia trachomatis inclusions.

    OpenAIRE

    Chiappino, M L; Dawson, C; Schachter, J; Nichols, B A

    1995-01-01

    The origin and distribution of glycogen in inclusions of Chlamydia trachomatis were demonstrated with silver proteinate stain for electron microscopy. Glycogen particles were detected in all developmental stages of C. trachomatis, as well as free in the inclusions. Intrachlamydial glycogen was most common in elementary bodies but was also detected in intermediate forms and reticulate bodies (RBs). Abnormal divisions and breakdown of cytoplasmic membranes were common in RBs. Cytoplasmic conten...

  11. Linguistic Diversity and Social Inclusion

    Science.gov (United States)

    Piller, Ingrid; Takahashi, Kimie

    2011-01-01

    This introduction provides the framework for the special issue by describing the social inclusion agenda of neoliberal market democracies. While the social inclusion agenda has been widely adopted, social inclusion policies are often blind to the ways in which language proficiency and language ideologies mediate social inclusion in linguistically…

  12. Assessment of inclusive education

    DEFF Research Database (Denmark)

    Miller, Tanja

    2014-01-01

    The focus of this paper is on the way teachers, in teams can work with assessing the inclusive practice in their own classes. In 2012 a joint effort between CEPRA, teachers and school administrators from the municipality of Hjørring developed a dialog based model for continually assessing the qua...

  13. Mathematics Teaching and Inclusion

    DEFF Research Database (Denmark)

    This volume contains the proceedings of the 3rd Nordic Research Conference on Special Needs Education in Mathematics, which took place in Rebild organised by Aalborg University in November 23-25, 2005. The theme of the conference was Mathematics Education and Inclusion. The conference theme...

  14. Peer Tutoring for Inclusion

    Science.gov (United States)

    Temple, Viviene A.; Lynnes, Michelle D.

    2008-01-01

    Peer tutoring is a pedagogical technique that has promise to improve outcomes for students with a disability within existing resource constraints. Published empirically-based papers on peer-tutoring were descriptively analysed. Synthesis of these studies revealed that peer tutoring is effective in inclusive physical education contexts. Evidence…

  15. Penile Epidermal Inclusion Cyst

    Directory of Open Access Journals (Sweden)

    M. El-Shazly

    2012-01-01

    Full Text Available We report a case of epidermal inclusion cyst in a 32-year-old male. This was a complication of circumcision that was neglected over years to form stones and urethrocutaneous fistula. Complete excision of the cyst and repair of the fistula were performed successfully. Histopathological examination confirmed our diagnosis.

  16. Supporting Inclusive Growth

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

    msandilands

    2010-10-27

    Oct 27, 2010 ... United Nations Development Programme. WRC. Women's Rights and Citizenship ... The goal of the Supporting Inclusive Growth (SIG) program is to support rigorous and policy-oriented research that ... Exploring synergies with IDRC's Environmental Economics program and other lead players outside the ...

  17. Multilingualism and social inclusion

    NARCIS (Netherlands)

    Marácz, L.; Adamo, S.

    2017-01-01

    This is a thematic issue on the relation between multilingualism and social inclusion. Due to globalization, Europeanization, supranational and transnational regulations linguistic diversity and multilingualism are on the rise. Migration and old and new forms of mobility play an important role in

  18. COMPOSITION OF FOWLPOX VIRUS AND INCLUSION MATRIX.

    Science.gov (United States)

    RANDALL, C C; GAFFORD, L G; DARLINGTON, R W; HYDE, J

    1964-04-01

    Randall, Charles C. (University of Mississippi School of Medicine, Jackson), Lanelle G. Gafford, Robert W. Darlington, and James M. Hyde. Composition of fowlpox virus and inclusion matrix. J. Bacteriol. 87:939-944. 1964.-Inclusion bodies of fowlpox virus infection are especially favorable starting material for the isolation of virus and inclusion matrix. Electron micrographs of viral particles and matrix indicated a high degree of purification. Density-gradient centrifugation of virus in cesium chloride and potassium tartrate was unsatisfactory because of inactivation, and clumping or disintegration. Chemical analyses of virus and matrix revealed significant amounts of lipid, protein, and deoxyribonucleic acid, but no ribonucleic acid or carbohydrate. Approximately 47% of the weight of the virus and 83% of the matrix were extractable in chloroform-methanol. The lipid partitions of the petroleum ether extracts were similar, except that the phospholipid content of the matrix was 2.2 times that of the virus. Viral particles were sensitive to diethyl ether and chloroform.

  19. Inclusive differentiated instruction

    Directory of Open Access Journals (Sweden)

    Jerković Ljiljana S.

    2017-01-01

    Full Text Available Inclusive differentiated instruction is a new model of didactic instruction, theoretically described and established in this paper for the first time, after being experimentally verified through teaching of the mother tongue (instruction in reading and literature. Inclusive individually planned instruction is based on a phenomenological and constructivist didactic instructional paradigm. This type of teaching is essentially developmental and person-oriented. The key stages of inclusive differentiated instruction of literature are: 1 recognition of individual students' potential and educational needs regarding reading and work on literary texts; 2 planning and preparation of inclusive individually planned instruction in reading and literature; 3 actual class teaching of lessons thus prepared; and 4 evaluation of the student achievement following inclusive differentiated instruction in reading and literature. A highly important element of the planning and preparation of inclusive differentiated instruction is the creation of student profiles and inclusive individualized syllabi. Individualized syllabi specify the following: 1. a brief student profile; 2. the student position on the continuum of the learning outcomes of instruction in the Serbian language; 3. reverse-engineered macro-plan stages of instruction in the Serbian language (3.1. identifying expected outcomes and fundamental qualities of learners' work, 3.2. defining acceptable proofs of their realisation, 3.3. planning learning and teaching experiences, and 3.4. providing material and technical requisites for teaching; 4 the contents and procedure of individualized lessons targeting the student; 5 a plan of syllabus implementation monitoring and evaluation. The continuum of the learning outcomes of inclusive differentiated instruction in literature exists at three main levels, A, B and C. The three levels are: A reading techniques and learning about the main literary theory concepts; B

  20. Multilingualism and Social Inclusion

    DEFF Research Database (Denmark)

    2017-01-01

    This is a thematic issue on the relation between multilingualism and social inclusion. Due to globalization, Europeanization, supranational and transnational regulations linguistic diversity and multilingualism are on the rise. Migration and old and new forms of mobility play an important role...... into practice. This lead to forced linguistic assimilation and the elimination of cultural and linguistic heritage. Now, in the present age of globalization, linguistic diversity at the national state level has been recognized and multilingual states have been developing where all types of languages can be used...... in governance and daily life protected by a legal framework. This does not mean that there is full equality of languages. This carries over to the fair and just social inclusion of the speakers of these weaker, dominated languages as well. There is always a power question related to multilingualism. The ten...

  1. INTEGRATION-INCLUSION

    OpenAIRE

    KERAMITCHIEVSKA-BILJANOVSKA Vesna; MITREVSKA Roberta; SABOLICH Nada

    2015-01-01

    By defining the terms, we would like to show that mentally disabled children cannot follow the regular educational process, in the same way children without learning disabilities can.The process of rehabilitation and education of disabled persons with different kind of disabilities, enables independent social integration and good quality of living in our country in the last 50 years; throw there is no need of changes in the inclusive education.

  2. BACTERIAL PLASMIDS

    Directory of Open Access Journals (Sweden)

    Marina Dinic

    2007-12-01

    Full Text Available Plasmids, extrachromosomal DNA, were identified in bacteria pertaining to family of Enterobacteriacae for the very first time. After that, they were discovered in almost every single observed strain. The structure of plasmids is made of circular double chain DNA molecules which are replicated autonomously in a host cell. Their length may vary from few up to several hundred kilobase (kb. Among the bacteria, plasmids are mostly transferred horizontally by conjugation process. Plasmid replication process can be divided into three stages: initiation, elongation, and termination. The process involves DNA helicase I, DNA gyrase, DNA polymerase III, endonuclease, and ligase.Plasmids contain genes essential for plasmid function and their preservation in a host cell (the beginning and the control of replication. Some of them possess genes whichcontrol plasmid stability. There is a common opinion that plasmids are unnecessary fora growth of bacterial population and their vital functions; thus, in many cases they can be taken up or kicked out with no lethal effects to a plasmid host cell. However,there are numerous biological functions of bacteria related to plasmids. Plasmids identification and classification are based upon their genetic features which are presented permanently in all of them, and these are: abilities to preserve themselves in a host cell and to control a replication process. In this way, plasmids classification among incompatibility groups is performed. The method of replicon typing, which is based on genotype and not on phenotype characteristics, has the same results as in compatibility grouping.

  3. Inclusive Education in Italy: Description and Reflections on Full Inclusion

    Science.gov (United States)

    Anastasiou, Dimitris; Kauffman, James M.; Di Nuovo, Santo

    2015-01-01

    Inclusion of students with disabilities when appropriate is an important goal of special education for students with special needs. Full inclusion, meaning no education for any child in a separate setting, is held to be desirable by some, and Italy is likely the nation with an education system most closely approximating full inclusion on the…

  4. High prevalence of biofilm synergy among bacterial soil isolates in cocultures indicates bacterial interspecific cooperation

    DEFF Research Database (Denmark)

    Ren, Dawei; Madsen, Jonas Stenløkke; Sørensen, Søren Johannes

    2015-01-01

    of single-species biofilms, indicating that all the individual strains benefit from inclusion in the multispecies community. Our results show a high prevalence of synergy in biofilm formation in multispecies consortia isolated from a natural bacterial habitat and suggest that interspecific cooperation...

  5. Supporting Teachers in Inclusive Education

    Directory of Open Access Journals (Sweden)

    Alekhina S.V.

    2015-03-01

    Full Text Available The article regards the issues of support provision to teachers involved in inclusive education as the main requirement for successful realization of inclusion. The methodological framework used in the study is a resource approach. The article describes the ways of extending the means of supporting teachers. The article also arguments for consolidating all the educators of inclusive schools into inclusive teams equally interested in joint work of administration and educators of intervention programs.

  6. Designing Inclusive Spaces

    DEFF Research Database (Denmark)

    Colfelt, Solvej

    2012-01-01

    CWUAAT -6.TH CAMBRIDGE WORKSHOP – 2012 Designing inclusive systems for real-world applications Abstracht: Denmark has planned huge investments in development in healthcare systems. Nearly 50 billion danish krones has been set aside on the stately budget for this purpose to be spent over the next 10...... hospital complexes ? The article will explore the fundament of wayshowing on the basis of prior research as well as on the basis of the results of a case study in a large existing danish hospital complex. The result points to signage being an inevitable factor but also that it is a factor that is not very...

  7. Inclusive Education under Collectivistic Culture

    Science.gov (United States)

    Futaba, Yasuko

    2016-01-01

    This paper addresses how inclusive education under collective culture is possible. Inclusive education, which more-or-less involves changing the current schools, has been denied, doubted or distorted by both policy-makers and practitioners of general and special education in Japan. Main reason for the setback in inclusive education can be…

  8. Inclusive Education in South Korea

    Science.gov (United States)

    Kim, Yong-Wook

    2014-01-01

    The purpose of this paper is to examine the current implementation of inclusive education in South Korea and discuss its challenges. The history of special education is first described followed by an introduction to policies relevant to special and inclusive education. Next, a critical discussion of the state of inclusive education follows built…

  9. Financial inclusion: Policies and practices

    Directory of Open Access Journals (Sweden)

    Thankom Arun

    2015-12-01

    Full Text Available As a key enabler for development, financial inclusion is firmly placed on the agenda of most governments as a key policy priority. Against this background, this round table provides a global and regional perspective on the policies and practices of financial inclusion. Using macro data, the collection reveals the diversity in the efforts towards achieving financial inclusion and the need for a progressive approach in financial inclusion. Further to this, the round table provides the regional perspectives on the policies and practices of financial inclusion in India, South Africa, and Australia.

  10. Inclusive Physical Education

    DEFF Research Database (Denmark)

    Østergaard, Charlotte; Rostbøll, Solveig Fogh

    2015-01-01

    EN317 - Inclusive Physical Education - with a focus on active and successful participation Charlotte Østergaard, Solveig Fogh Rostbøll, Department of School and Learning, Metropolitan University College (DK) chao@phmetropol.dk The Danish School Reform 2014 intends to raise the amount and intensity...... and ability to participate in PE must be understood in specific socio-cultural and socio-economic conditions. The hypothesis of the study is that the experience of being acknowledged for your efforts in physical education by significant others can form the basis for the construction of physical capital. EN323...... - Iranian Female Adolescent Immigrants’ Experiences of the Canadian Physical Education System Bahar Tajrobehkar, University of Toronto (CA) btajro@gmail.co...

  11. Can we build inclusion?

    DEFF Research Database (Denmark)

    Kirkeby, Inge Mette; Grangaard, Sidse

    2016-01-01

    of some children with special needs. Hence the title – can we build inclusion? In the literature of Universal Design, accommodation and design features seldom reflect the less visible disabilities. The paper is based on a research project initiated to investigate how more or less space influences daily......, the amount of space is crucial. This group consisted of children who were socially very extrovert, and who maybe were noisy, easily provoked, and quick to get involved in arguments with other children. Alternatively, children in the group were very restrained and withdrawn in social interaction. Based...... and furnishing of the kindergarten is an advantage for all children. This is a clear example of Universal Design in which architectural solutions that are good for one specific group are also beneficial for all children...

  12. Between psychopathology and inclusion

    DEFF Research Database (Denmark)

    Hamre, Bjørn Frithiof; Hedegaard-Sørensen, Lotte; Langager, Søren

    2017-01-01

    and social pedagogy. We thus enquire into how the rise of diagnostics and medicalisation affects our understanding of children’s difficulties. We discuss a paradox that is present in Denmark and other countries. As educational policies emphasise inclusion, the field of schooling experiences a huge rise......This article reports on a Danish study on interprofessional collaboration between child psychiatrists and educational psychologists concerning children who are categorised as being at risk. Methodologically, the analysis is grounded in qualitative interviews with psychologists. A Foucauldian...... approach is applied to narratives and experiences that occur within these interviews concerning external collaboration with child psychiatrists. The article is informed by the research tradition that has problematised the significance of psychiatry and diagnoses in the field of special needs education...

  13. IPads in Inclusive Classrooms

    DEFF Research Database (Denmark)

    Meyer, Bente Tobiesen

    2015-01-01

    This paper builds on data from a research project where iPads were used in a lower secondary school in Denmark to support school development and inclusive learning environments. The paper explores how iPads enter into and work as part of an ecology of learning resources in five classes in lower...... secondary school. I conceptualize the systems of related technologies observed in this school as ecologies of learning resources as they present themselves as carefully balanced systems in which educational resources circulate in different ways that make sense for learners’ needs. Inspired by Actor...... in by pupils the paper argues that we should disengage approaches to the iPad in education from ideas of what the properties of these technologies are, and see the device as a more relational and situated actor, avoiding the definition of properties of technologies outside the contexts specific to their use....

  14. Patterns of inclusion

    DEFF Research Database (Denmark)

    Pedersen, Alex Young; Nørgård, Rikke Toft; Köppe, Christian

    2018-01-01

    Reconsidering the concept of digital citizenship and the essential component of education the authors propose that the concept of Hybrid Education may serve both as a guideline for the utilization of digital technologies in education and as a methodology for fostering new forms of participation......, inclusion and engagement in society. Following T.H. Marshall’s conception of citizenship the authors suggest that becoming, belonging and the capabilities to do so is essential to digital citizenship in a culturally diverse and digitally mediated world. The paper presents a theory-based, value driven...... and practical orientated framework for innovation in education. The paper documents a process where participants situated around the globe worked in collaboration with a group of participants gathered at a workshop on the concept of hybridity in education and on identifying and describing educational patterns...

  15. Inclusive Business - What It Is All About? Managing Inclusive Companies

    Directory of Open Access Journals (Sweden)

    Tea Golja

    2012-01-01

    Full Text Available Following the challenges we face today, the inclusive business models are future business models through which the Millennium Development Goals can be fostered and strengthen. These are the models which, through their strategic orientation on inclusivity, include low income communities in their value chain. This can be done through combining variety of strategies which all have two common points – recognition of stakeholders and adjustment of the product to the target market. The paper presents the analysis of inclusive markets. Hence, the research results show the dispersion of inclusive businesses worldwide, type of the organization, sector coverage, and contribution to MDGs as well as the particular way of inclusion of low income communities in their value chain. The aim is to present how inclusive business benefits not only the low income societies, but the companies that operate in this way as well.

  16. Inclusive spectra in hard processes

    International Nuclear Information System (INIS)

    Kiselev, A.V.; Petrov, V.A.

    1985-01-01

    It is shown that the unified mechanism of hadronization in hard processes results in universality of inclusive spectra of soft hadrons. Inclusive spectrum of hadrons in energy share in deep-inelastic lepton-hadron scattering is calculated. The spectrum obtained is calculated with analogous distribution in e + e - annihilation. It is noted that inclusive spectrum of soft hadrons in hard processes is described by a universal function

  17. Financial Stability and Financial Inclusion

    OpenAIRE

    Peter J. Morgan; Victor Pontines

    2014-01-01

    Developing economies are seeking to promote financial inclusion, i.e., greater access to financial services for low-income households and firms, as part of their overall strategies for economic and financial development. This raises the question of whether financial stability and financial inclusion are, broadly speaking, substitutes or complements. In other words, does the move toward greater financial inclusion tend to increase or decrease financial stability? A number of studies have sugge...

  18. Accessibility and inclusion informational

    Directory of Open Access Journals (Sweden)

    Mônica Sena de Souza

    2013-06-01

    Full Text Available Introduction: Discusses the role of information professionals in meeting the informational demands of people with disabilities in the information society. The librarian is crucial for the effectiveness and success in the informational inclusion of people with disabilities, considering also continuing education for their professional qualification.Objective: To provide reflections on the role of the librarian in serving users with disabilities, highlighting the need for improvement in information units, identified in the scientific literature with regard to accessibility.Methodology: Literature search, based on a review of literature in books and scientific papers, highlighting the main authors: Adams (2000, Mazzoni (2001 and Sassaki (1997, 2002, 2005.Results: The lack of informational access for people with disabilities hampers their social and political participation, hence, reduces its condition of citizenship.Conclusion: The librarian responsible for seeking continuing education, greater involvement in the events of the area and the constant search for job training, which will reflect on the best service the information needs of users with disabilities.

  19. Inclusion by Design: Engineering Inclusive Practices in Secondary Schools

    Science.gov (United States)

    Dukes, Charles; Lamar-Dukes, Pamela

    2009-01-01

    In order to help teachers understand the importance of intentional design for inclusive education, this article describes the design process an engineer might use when designing a new project. If teachers learn to think like engineers, it is possible for them to design inclusive education. This conceptual design can then be combined with…

  20. Designing Inclusive Systems Designing Inclusion for Real-world Applications

    CERN Document Server

    Clarkson, John; Robinson, Peter; Lazar, Jonathan; Heylighen, Ann

    2012-01-01

    The Cambridge Workshops on Universal Access and Assistive Technology (CWUAAT) are a series of workshops held at a Cambridge University College every two years. The workshop theme: “Designing inclusion for real-world applications” refers to the emerging potential and relevance of the latest generations of inclusive design thinking, tools, techniques, and data, to mainstream project applications such as healthcare and the design of working environments. Inclusive Design Research involves developing tools and guidance enabling product designers to design for the widest possible population, for a given range of capabilities. There are five main themes: •Designing for the Real-World •Measuring Demand And Capabilities •Designing Cognitive Interaction with Emerging Technologies •Design for Inclusion •Designing Inclusive Architecture In the tradition of CWUAAT, we have solicited and accepted contributions over a wide range of topics, both within individual themes and also across the workshop’s scope. ...

  1. The clinical impact of bacterial biofilms

    DEFF Research Database (Denmark)

    Høiby, Niels; Ciofu, Oana; Johansen, Helle Krogh

    2011-01-01

    . Likewise, chronic Pseudomonas aeruginosa lung infections in cystic fibrosis patients are caused by biofilm growing mucoid strains. Gradients of nutrients and oxygen exist from the top to the bottom of biofilms and the bacterial cells located in nutrient poor areas have decreased metabolic activity....... Bacterial biofilms are resistant to antibiotics, disinfectant chemicals and to phagocytosis and other components of the innate and adaptive inflammatory defense system of the body. It is known, for example, that persistence of staphylococcal infections related to foreign bodies is due to biofilm formation...

  2. Bacterial Cholangitis, Cholecystitis, or both in Dogs.

    Science.gov (United States)

    Tamborini, A; Jahns, H; McAllister, H; Kent, A; Harris, B; Procoli, F; Allenspach, K; Hall, E J; Day, M J; Watson, P J; O'Neill, E J

    2016-07-01

    Bacterial cholangitis and cholecystitis are rarely reported, poorly characterized diseases in the dog. To characterize the clinical features of these conditions. Twenty-seven client-owned dogs with bacterial cholangitis, cholecystitis, or both. Multicenter, retrospective cases series of dogs with bacterial cholangitis, cholecystitis, or both, presenting January 2000 to June 2011 to 4 Veterinary Schools in Ireland/United Kingdom. Interrogation of hospital databases identified all cases with the inclusion criteria; histopathologically confirmed cholangitis or cholecystitis and bile culture/cytology results supporting a bacterial etiology. Twenty-seven dogs met the inclusion criteria with approximately 460 hepatitis cases documented over the same study period. Typical clinical pathology findings were increases in liver enzyme activities (25/26), hyperbilirubinemia (20/26), and an inflammatory leukogram (21/24). Ultrasound findings, although nonspecific, aided decision-making in 25/26 cases. The most frequent hepatobiliary bacterial isolates were Escherichia coli (n = 17; 16 cases), Enterococcus spp. (n = 8; 6 cases), and Clostridium spp. (n = 5; 5 cases). Antimicrobial resistance was an important feature of aerobic isolates; 10/16 E. coli isolates resistant to 3 or more antimicrobial classes. Biliary tract rupture complicated nearly one third of cases, associated with significant mortality (4/8). Discharged dogs had a guarded to fair prognosis; 17/18 alive at 2 months, although 5/10 re-evaluated had persistent liver enzyme elevation 2-12 months later. Bacterial cholangitis and cholecystitis occur more frequently than suggested by current literature and should be considered in dogs presenting with jaundice and fever, abdominal pain, or an inflammatory leukogram or with ultrasonographic evidence of gallbladder abnormalities. Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of

  3. Creative Educational Practices for Inclusion

    Science.gov (United States)

    Piske, Fernanda Hellen Ribeiro; Stoltz, Tania; Machado, Jarci

    2014-01-01

    Inclusion of gifted students depends on several aspects to happen in the school context, and one of the most important aspects to include these children at school is creative educational practices. Teaching with art is a good possibility to make children feel motivated to attend school. In the school context, the inclusion of these children could…

  4. Early Childhood Inclusion in Turkey

    Science.gov (United States)

    Diken, Ibrahim H.; Rakap, Salih; Diken, Ozlem; Tomris, Gozde; Celik, Secil

    2016-01-01

    Inclusion of young children with disabilities into regular preschool classrooms is a common practice that has been implemented for several decades in industrialized nations around the world, and many developing countries including Turkey have been developing and implementing laws, regulation, and services to support inclusion and teaching in…

  5. Inclusion in Malaysian Integrated Preschools

    Science.gov (United States)

    Sukumaran, Sailajah; Loveridge, Judith; Green, Vanessa A.

    2015-01-01

    Inclusive education has been introduced through a number of policy developments in Malaysia over the last 10 years but there is little research investigating the extent and nature of inclusive education for preschoolers with special educational needs (SEN). This study surveyed both regular and special education teachers in Malaysian integrated…

  6. Stiffening solids with liquid inclusions

    Science.gov (United States)

    Style, Robert W.; Boltyanskiy, Rostislav; Allen, Benjamin; Jensen, Katharine E.; Foote, Henry P.; Wettlaufer, John S.; Dufresne, Eric R.

    2015-01-01

    From bone and wood to concrete and carbon fibre, composites are ubiquitous natural and synthetic materials. Eshelby’s inclusion theory describes how macroscopic stress fields couple to isolated microscopic inclusions, allowing prediction of a composite’s bulk mechanical properties from a knowledge of its microstructure. It has been extended to describe a wide variety of phenomena from solid fracture to cell adhesion. Here, we show experimentally and theoretically that Eshelby’s theory breaks down for small liquid inclusions in a soft solid. In this limit, an isolated droplet’s deformation is strongly size-dependent, with the smallest droplets mimicking the behaviour of solid inclusions. Furthermore, in opposition to the predictions of conventional composite theory, we find that finite concentrations of small liquid inclusions enhance the stiffness of soft solids. A straightforward extension of Eshelby’s theory, accounting for the surface tension of the solid-liquid interface, explains our experimental observations. The counterintuitive stiffening of solids by fluid inclusions is expected whenever inclusion radii are smaller than an elastocapillary length, given by the ratio of the surface tension to Young’s modulus of the solid matrix. These results suggest that surface tension can be a simple and effective mechanism to cloak the far-field elastic signature of inclusions.

  7. Early Childhood Inclusion in Spain

    Science.gov (United States)

    Giné, Climent; Balcells-Balcells, Anna; Cañadas, Margarita; Paniagua, Gema

    2016-01-01

    This article describes early childhood inclusion in educational settings in Spain. First, we address the legislative framework of preschool education in Spain and offer a brief analysis of some relevant issues, including the current situation of early childhood education and inclusion at this stage. Second, current policies and practices relating…

  8. Social Inclusion and Metrolingual Practices

    Science.gov (United States)

    Otsuji, Emi; Pennycook, Alastair

    2011-01-01

    In this paper, we explore the implications of metrolingual language practices for how we understand social inclusion. A vision of social inclusion that includes bi- and multilingual capacities may comprise an appreciation of a diversity of languages other than English, and the skills and capabilities of multilingual language users, yet it is all…

  9. Pollution monitoring of coastal and estuarine areas: 1. Bacterial indicators along the south Gujarat Coast

    Digital Repository Service at National Institute of Oceanography (India)

    Mohandass, C.; Nair, S.; Achuthankutty, C.T.; LokaBharathi, P.A.

    . Total coliforms and total vibrios were retrieved in 27 and 55 % of the samples. The validity of these bacterial groups as water quality indicators is discussed with statistical relations. It is sugested that inclusion of these groups could be made...

  10. The infidelity of melt inclusions?

    Science.gov (United States)

    Kent, A. J.

    2008-12-01

    Melt inclusions provide important information about magmatic systems and represent unique records of magma composition and evolution. However, it is also clear that melt inclusions do not necessarily constitute a petrological 'magic bullet', and potential exists for trapped melt compositions to be modified by a range of inclusion-specific processes. These include trapping of diffusional boundary layers, crystallization of the host mineral after trapping and dissolution of co-trapped minerals during homogenization, diffusional exchange between trapped liquid and the host mineral and external melt, and cryptic alteration of trapped material during weathering or hydrothermal alteration. It clearly important to identify when melt inclusions are unmodified, and which compositional indices represent the most robust sources of petrogenetic information. In this presentation I review and discuss various approaches for evaluating compositions and compositional variations in inclusion suites. An overriding principle is that the variations evident in melt inclusions should be able to be understood in terms of petrological processes that are known, or can be reasonably inferred to also effect bulk magma compositions. One common approach is to base petrological conclusions on species that should be more robust, and many workers use variations in incompatible trace elements for this purpose. However important information may also be obtained from a comparison of variations in melt inclusions and the lavas that host them, and in most cases this comparison is the key to identifying inclusions and suites that are potentially suspect. Comparisons can be made between individual inclusions and lavas, although comparison of average inclusion composition and the host lava, after correction for differences in crystal fractionation, may also be valuable. An important extension of this is the comparison of the variability of different species in inclusions and host lavas. This also provides

  11. Prevention of bacterial adhesion

    DEFF Research Database (Denmark)

    Klemm, Per; Vejborg, Rebecca Munk; Hancock, Viktoria

    2010-01-01

    Management of bacterial infections is becoming increasingly difficult due to the emergence and increasing prevalence of bacterial pathogens that are resistant to available antibiotics. Conventional antibiotics generally kill bacteria by interfering with vital cellular functions, an approach...... that imposes selection pressure for resistant bacteria. New approaches are urgently needed. Targeting bacterial virulence functions directly is an attractive alternative. An obvious target is bacterial adhesion. Bacterial adhesion to surfaces is the first step in colonization, invasion, and biofilm formation....... As such, adhesion represents the Achilles heel of crucial pathogenic functions. It follows that interference with adhesion can reduce bacterial virulence. Here, we illustrate this important topic with examples of techniques being developed that can inhibit bacterial adhesion. Some of these will become...

  12. Ca isotopes in refractory inclusions

    International Nuclear Information System (INIS)

    Niederer, F.R.; Papanastassiou, D.A.

    1984-01-01

    We report measurements of the absolute isotope abundance of Ca in Ca-Al-rich inclusions from the Allende and Leoville meteorites. Improved high precision measurements are reported also for 46 Ca. We find that nonlinear isotope effects in Ca are extremely rare in these inclusions. The absence of nonlinear effects in Ca, except for the effects in FUN inclusions, is in sharp contrast to the endemic effects in Ti. One fine-grained inclusion shows an excess of 46 Ca of (7 +- 1) per mille, which is consistent with addition of only 46 Ca or of an exotic (*) component with 46 Ca* approx. 48 Ca*. FUN inclusion EK-1-4-1 shows a small 46 Ca excess of (3.3 +- 1.0) per mille; this confirms that the exotic Ca components in EK-1-4-1 were even more deficient in 46 Ca relative to 48 Ca than is the case for normal Ca. The Ca in the Ca-Al-rich inclusions shows mass dependent isotope fractionation effects which have a range from -3.8 to +6.7 per mille per mass unit difference. This range is a factor of 20 wider than the range previously established for bulk meteorites and for terrestrial and lunar samples. Ca and Mg isotope fractionation effects in the Ca-Al-rich inclusions are common and attributed to kinetic isotope effects. (author)

  13. Inclusive education and social exclusion

    Directory of Open Access Journals (Sweden)

    Maria Luisa Bissoto

    2013-01-01

    Full Text Available The aim of this paper is critically examining assumptions underlying the Inclusive Education concept, arguing that this can only be effectively considered when understood in a broader context of social inclusion and exclusion. Methodologically, this article relies on international documents and bibliographic references about Inclusive Education, that have been chosen by systematize and characterize different social and educational inclusive practices, encouraging the elaboration of a general overview on this topic. The results of this analysis conclude that it is essential for Inclusive Education that educational institutions review their goals and reasons of social existence. In the concluding remarks it is argued that education is better understood as the act of encouraging and welcoming the efforts of individuals in their attempts to engage in social networking, which sustains life. This includes the acceptance of other reality interpretations and understanding that educational action cannot be restricted by the walls of institutions. It requires the participation of the whole community. Action perspectives likely to promote social inclusion and inclusive education are suggested.

  14. Molecular basis for dominantly inherited inclusion body β-thalassemia

    International Nuclear Information System (INIS)

    Thein, S.L.; Hesketh, C.; Wood, W.G.; Clegg, J.B.; Old, J.M.; Weatherall, D.J.; Taylor, P.; Temperley, I.J.; Hutchinson, R.M.

    1990-01-01

    Analysis of the molecular basis of dominantly inherited β-thalassemia in four families has revealed different mutations involving exon 3 of the β-globin gene. It is suggested that the phenotypic difference between this condition and the more common recessive forms of β-thalassemia lies mainly in the length and stability of the abnormal translation products that are synthesized and, in particular, whether they are capable of binding heme and producing aggregations that are relatively resistant to proteolytic degradation

  15. Inclusion bodies from recombinant bacteria as antigen to directly ...

    African Journals Online (AJOL)

    Jane

    2011-08-03

    Aug 3, 2011 ... chain reaction (TD-PCR) condition was used to amplify the PLA2 gene following reverse transcription reaction with oligo d(T)18. A 460 bp of PCR product was visualized with 1% agarose ... The supernatant was slowly added with saturated ammonium sulfate to a concentration of 45% and stirred for 30 min.

  16. Electromagnetism of Bacterial Growth

    Science.gov (United States)

    Ainiwaer, Ailiyasi

    2011-10-01

    There has been increasing concern from the public about personal health due to the significant rise in the daily use of electrical devices such as cell phones, radios, computers, GPS, video games and television. All of these devices create electromagnetic (EM) fields, which are simply magnetic and electric fields surrounding the appliances that simultaneously affect the human bio-system. Although these can affect the human system, obstacles can easily shield or weaken the electrical fields; however, magnetic fields cannot be weakened and can pass through walls, human bodies and most other objects. The present study was conducted to examine the possible effects of bacteria when exposed to magnetic fields. The results indicate that a strong causal relationship is not clear, since different magnetic fields affect the bacteria differently, with some causing an increase in bacterial cells, and others causing a decrease in the same cells. This phenomenon has yet to be explained, but the current study attempts to offer a mathematical explanation for this occurrence. The researchers added cultures to the magnetic fields to examine any effects to ion transportation. Researchers discovered ions such as potassium and sodium are affected by the magnetic field. A formula is presented in the analysis section to explain this effect.

  17. Aquareovirus NS80 recruits viral proteins to its inclusions, and its C-terminal domain is the primary driving force for viral inclusion formation.

    Directory of Open Access Journals (Sweden)

    Ling Shao

    Full Text Available Cytoplasmic inclusion bodies formed in reovirus-infected cells are the sites of viral replication and assembly. Previous studies have suggested that the NS80 protein of aquareovirus may be involved in the formation of viral inclusion bodies. However, it remains unknown whether other viral proteins are involved in the process, and what regions of NS80 may act coordinately in mediating inclusion formation. Here, we observed that globular cytoplasmic inclusions were formed in virus-infected cells and viral proteins NS80 and NS38 colocalized in the inclusions. During transfection, singly expressed NS80 could form cytoplasmic inclusions and recruit NS38 and GFP-tagged VP4 to these structures. Further treatment of cells with nocodazole, a microtubule inhibitor, did not disrupt the inclusion, suggesting that inclusion formation does not rely on microtubule network. Besides, we identified that the region 530-742 of NS80 was likely the minimal region required for inclusion formation, and the C-tail, coiled-coil region as well as the conserved linker region were essential for inclusion phenotype. Moreover, with series deletions from the N-terminus, a stepwise conversion occurred from large condensed cytoplasmic to small nuclear inclusions, then to a diffused intracellular distribution. Notablely, we found that the nuclear inclusions, formed by NS80 truncations (471 to 513-742, colocalized with cellular protein β-catenin. These data indicated that NS80 could be a major mediator in recruiting NS38 and VP4 into inclusion structures, and the C-terminus of NS80 is responsible for inclusion formation.

  18. Politics of Inclusion and Empowerment

    DEFF Research Database (Denmark)

    Andersen, John; Siim, Birte

    2004-01-01

    The objective of the book is to analyse different politics of inclusion and empowerment and the different paradigms of inclusion/exclusion in order to underline the close link between politics of scoial equality and politics of recognition of ciultural difference. Politics of inclusion is thus...... theproductive/innovative linkage of politics of redistributuin and politics og resognition, whnich over a longer time span creates sustainable paths of democratic and social development, which increases the capacity to handle both conflicts about economic resources and life-chances and conflicts about...

  19. Measuring Nurse Educators' Willingness to Adopt Inclusive Teaching Strategies.

    Science.gov (United States)

    Levey, Janet A

    The purpose of the study was to examine the characteristics and relationships of nurse educators' teaching practices, knowledge, support, and willingness to adopt inclusive teaching strategies (WillAdITS). Adopting more inclusive teaching strategies based on universal design for instruction is an innovative way for educators to reach today's diverse student body. However, the pedagogy has not diffused into nursing education. Descriptive statistics and hierarchical multiple regression were used for analyzing data from 311 nurse educators in prelicensure and RN to BSN programs. The model explained 44.8 percent of the variance in WillAdITS. The best indicators for this pedagogy were knowledge of universal design for instruction, social system support for inclusive teaching strategies, multiple instructional formats, and years of teaching. Knowing factors influencing the adoption of inclusive teaching strategies can inform schools of nursing of areas needing further development in the preparation of novice to experienced educators to teach diverse learners.

  20. Inclusion and exclusion strategies for conducting meta-analyses.

    Science.gov (United States)

    Swift, Joshua K; Wampold, Bruce E

    2018-05-01

    Some of the most important decisions that a researcher will make when conducting a meta-analysis are decisions about the search strategies and inclusion/exclusion criteria. Decisions regarding inclusion/exclusion criteria serve to define the scope of a meta-analysis and search strategy decisions can have a large impact on how well the results of a meta-analysis actually represent the existing body of literature. In this article, we provide descriptions and recommendations for performing searches and making inclusion/exclusion decisions. We also provide a review of 65 meta-analyses of psychotherapy versus pharmacotherapy in order to offer frequency data on the use of various search strategies and inclusion/exclusion decisions in the field.

  1. Inclusão e governamentalidade Inclusion and governmentality

    Directory of Open Access Journals (Sweden)

    Alfredo Veiga-Neto

    2007-10-01

    Full Text Available O artigo parte dos conceitos de governamento, biopoder, norma e governamentalidade para argumentar no sentido de que as políticas de inclusão - especialmente as políticas de inclusão escolar - são dispositivos biopolíticos para o governamento e o controle das populações. Tomam-se esses e outros conceitos foucaultianos como ferramentas analíticas para argumentar que é preciso examinar as políticas e práticas inclusivas sem assumir antecipadamente qualquer juízo de valor sobre elas. Entendendo a diferença como manifestação da diversidade, e o diferente como um exótico, um portador de algo que os outros não possuem, as políticas de inclusão - nos modos como vêm sendo formuladas e em parte executadas no Brasil - parecem ignorar a própria diferença. Assim, ao invés de promoverem uma educação para todos, elas correm o risco de realizar uma inclusão excludente das diferenças. Discute-se a distinção entre campanhas públicas e políticas de inclusão em termos dos âmbitos em que ambas se efetivam.Based on the concepts of government, biopower, norm and governmentality, this paper argues that the inclusion policies - especially those of school inclusion - are biopolitical devices to govern and control the populations. These and other concepts by Foucault are used as analytical tools to argue that we have to explore the including policies and practices without any previous value judgment on them. Since they understand the difference as a manifestation of diversity, and the different as the exotic, what carries something the others do not have, the inclusion policies - as they are being formulated and partly enforced in Brazil - seem to ignore difference itself. Thus, instead of promoting education for all, they may lead to an inclusion that excludes the differences. The text finally discusses the distinction between public campaigns and inclusion policies in terms of the settings in which both take place.

  2. Inclusões intracitoplasmáticas hialinas na medular da adrenal de bovinos

    Directory of Open Access Journals (Sweden)

    L.P Mesquita

    2011-02-01

    Full Text Available Cytoplasmic inclusion bodies in adrenal medullary chromaffin cells have been described in various species including humans. These inclusions are believed to be related to certain infectious, toxic and neurodegenerative diseases. No reports concerning such adrenal inclusions have been described in bovines. Adrenal glands from twenty bovines were evaluated in a retrospective study. Seven of these exhibited inclusions - three cases of rabies, two cases of chronic suppurative bronchopneumonia, one case of chronic suppurative peritonitis, and one case of gangrenous mastitis. The inclusions were present in higher numbers especially in cases of rabies and also in one case of chronic suppurative bronchopneumonia. The inclusions were intracytoplasmic, eosinophilic, rounded, single or multiple, of various sizes, strongly stained by PAS and were present in higher numbers in the external layer of the adrenal medulla. The inclusions were negative when subjected to immunohistochemistry for detection of viral antigens in the cases of rabies. Although inclusion bodies were present in adrenal glands devoid of other histological alterations, they were more abundant in cases in which the adrenal gland had other alterations. The correlation between certain diseases and the development of inclusion bodies is not known, which highlights the importance of further studies on these inclusions in adrenal glands of bovines.

  3. Inclusion for Customer with Disabilities

    OpenAIRE

    Murphy, James Peter

    2010-01-01

    For many people with disabilities, licensed premises design and lack of sufficient staff training can act as powerful barriers to their full inclusion in the activities of bars, restaurants and nightclubs.

  4. EDUCATIONAL INCLUSION AND CRITICAL PEDAGOGY

    Directory of Open Access Journals (Sweden)

    Silvia Molina

    2009-11-01

    Full Text Available This article outlines the basic principles of educational inclusion, focusingspecifically on the inclusion of disabled students in mainstream classrooms, and arguesthat inclusive education should be understood as a process of transforming traditionalschools into spaces of learning for all students. The article uses the lens of critical pedagogyto argue that exclusionary educational practices have been developed through themedicalization of learning disabilities which focused on the disability rather than theabilities of disabled students. Following the same line of thinking, the article providesscientific evidence to debunk myths related to the education of disabled students; especiallymyths that contributed to their exclusion from mainstream classrooms. Finally,based on the Learning Communities model, we provide some concrete strategies fortransforming mainstream classrooms into fully inclusive environments.

  5. PSYCHOANALYSIS, ETHICS AND INCLUSIVE EDUCATION

    OpenAIRE

    Ana Beatriz Coutinho Lerner; Rinaldo Voltolini

    2016-01-01

    This paper aims to investigate the central ways in which the discourse on Inclusive Education assumes in contemporary times and some the effects of this discourse on the subjectivity of children, parents and school professionals. In order to accomplish this goal, we first review the literature concerning Inclusive Education, showing the origins of this movement, the main legal frameworks that organize this subject and the current theoretical perspectives. We then analyze discursive elements c...

  6. Forum, Dedicated to Inclusive Education

    Directory of Open Access Journals (Sweden)

    Vachkov I.V.

    2015-06-01

    Full Text Available 26 – 27 of February 2015 in Kazan, in the University of Management “TISBI” been held National (All-Russian forum of promotion of ideas and principles of inclusive education (with international participants “Study and live together: open space of inclusion”. During the work of Forum the most topical questions of inclusive education implement in Russian Federation been discussed.

  7. Studying bacterial multispecies biofilms

    DEFF Research Database (Denmark)

    Røder, Henriette Lyng; Sørensen, Søren Johannes; Burmølle, Mette

    2016-01-01

    , but the identity and significance of interspecies bacterial interactions is neglected in these analyses. There is therefore an urgent need for bridging the gap between metagenomic analysis and in vitro models suitable for studies of bacterial interactions.Bacterial interactions and coadaptation are important......The high prevalence and significance of multispecies biofilms have now been demonstrated in various bacterial habitats with medical, industrial, and ecological relevance. It is highly evident that several species of bacteria coexist and interact in biofilms, which highlights the need for evaluating...

  8. Body Hair

    Science.gov (United States)

    ... girlshealth.gov/ Home Body Puberty Body hair Body hair Even before you get your first period , you ... removing pubic hair Ways to get rid of hair top Removing body hair can cause skin irritation, ...

  9. Nile Red Detection of Bacterial Hydrocarbons and Ketones in a High-Throughput Format

    Energy Technology Data Exchange (ETDEWEB)

    Pinzon, NM; Aukema, KG; Gralnick, JA; Wackett, LP

    2011-06-28

    A method for use in high-throughput screening of bacteria for the production of long-chain hydrocarbons and ketones by monitoring fluorescent light emission in the presence of Nile red is described. Nile red has previously been used to screen for polyhydroxybutyrate (PHB) and fatty acid esters, but this is the first report of screening for recombinant bacteria making hydrocarbons or ketones. The microtiter plate assay was evaluated using wild-type and recombinant strains of Shewanella oneidensis and Escherichia coli expressing the enzyme OleA, previously shown to initiate hydrocarbon biosynthesis. The strains expressing exogenous Stenotrophomonas maltophilia oleA, with increased levels of ketone production as determined by gas chromatography-mass spectrometry, were distinguished with Nile red fluorescence. Confocal microscopy images of S. oneidensis oleA-expressing strains stained with Nile red were consistent with a membrane localization of the ketones. This differed from Nile red staining of bacterial PHB or algal lipid droplets that showed intracellular inclusion bodies. These results demonstrated the applicability of Nile red in a high-throughput technique for the detection of bacterial hydrocarbons and ketones. IMPORTANCE In recent years, there has been renewed interest in advanced biofuel sources such as bacterial hydrocarbon production. Previous studies used solvent extraction of bacterial cultures followed by gas chromatography-mass spectrometry (GC-MS) to detect and quantify ketones and hydrocarbons (Beller HR, Goh EB, Keasling JD, Appl. Environ. Microbiol. 76: 1212-1223, 2010; Sukovich DJ, Seffernick JL, Richman JE, Gralnick JA, Wackett LP, Appl. Environ. Microbiol. 76: 3850-3862, 2010). While these analyses are powerful and accurate, their labor-intensive nature makes them intractable to high-throughput screening; therefore, methods for rapid identification of bacterial strains that are overproducing hydrocarbons are needed. The use of high

  10. Becoming Inclusive: A Code of Conduct for Inclusion and Diversity.

    Science.gov (United States)

    Schmidt, Bonnie J; MacWilliams, Brent R; Neal-Boylan, Leslie

    There are increasing concerns about exclusionary behaviors and lack of diversity in the nursing profession. Exclusionary behaviors, which may include incivility, bullying, and workplace violence, discriminate and isolate individuals and groups who are different, whereas inclusive behaviors encourage diversity. To address inclusion and diversity in nursing, this article offers a code of conduct. This code of conduct builds on existing nursing codes of ethics and applies to nursing students and nurses in both educational and practice settings. Inclusive behaviors that are demonstrated in nurses' relationships with patients, colleagues, the profession, and society are described. This code of conduct provides a basis for measureable change, empowerment, and unification of the profession. Recommendations, implications, and a pledge to action are discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Social inclusion and mental health.

    Science.gov (United States)

    Cobigo, Virginie; Stuart, Heather

    2010-09-01

    Recent research on approaches to improving social inclusion for people with mental disabilities is reviewed. We describe four approaches (or tools) that can be used to improve social inclusion for people with mental disabilities: legislation, community-based supports and services, antistigma/antidiscrimination initiatives, and system monitoring and evaluation. While legislative solutions are the most prevalent, and provide an important framework to support social inclusion, research shows that their full implementation remains problematic. Community-based supports and services that are person-centered and recovery-oriented hold considerable promise, but they are not widely available nor have they been widely evaluated. Antistigma and antidiscrimination strategies are gaining in popularity and offer important avenues for eliminating social barriers and promoting adequate and equitable access to care. Finally, in the context of the current human rights and evidence-based health paradigms, systematic evidence will be needed to support efforts to promote social inclusion for people with mental disabilities, highlight social inequities, and develop best practice approaches. Tools that promote social inclusion of persons with mental disabilities are available, though not yet implemented in a way to fully realize the goals of current disability discourse.

  12. Demands from the school inclusion

    Directory of Open Access Journals (Sweden)

    Selma Norberto Matos

    2014-04-01

    Full Text Available From the implementation of public policies on school inclusion, mainly those directed to the target audience of special education, the number of students with special educational needs in common classes has increased. This fact has helped to compose the picture in schools where the limitations and contradictions of the Brazilian educational system have appeared. Educational actors and authors are challenged to build knowledge able of responding to demands of daily school, concerning living and learning in diversity. Whereas this inclusive process is new in the schools, the study aimed to analyze the demands of teachers from the school inclusion. The research was qualitative and exploratory, and six teachers, their students with special educational needs and three professionals in the Nucleus of Inclusive Education from the Municipal Department of Education took in it. Technique of participant observation, field diary, semi-structured interview and questionnaire were used for data collection, while analysis of content was used for discussion of the data. The results indicate that there are achievements and contradictions in the reality of schools that themselves propose inclusive; advances and limitations resulting from the municipal politics; that the model of performance of the group of special education, in the context analyzed, may be revised or expanded; and that the teachers has demands with regard to public policy, training, and the psychologist.

  13. Exploring the Quality Indicators of a Successful Full-Inclusion Preschool Program

    Science.gov (United States)

    Warren, Susan R.; Martinez, Richard S.; Sortino, Lori A.

    2016-01-01

    A growing body of research and legislative policies support the importance of high-quality early intervention systems for preschool children with disabilities. Inclusion programs are viable means for providing this support, yet limited progress has been made in the past decade to increase the placements of children in inclusive settings or define…

  14. The Sociology of Social Inclusion

    Directory of Open Access Journals (Sweden)

    Dan Allman

    2013-01-01

    Full Text Available This article looks at social inclusion from a sociological perspective. It argues that sociology complements biological and other natural order explanations of social stratification. The article interrogates a variety of forms of social integration, including ostracism within 5th century b.c. Greece, 19th-century solidarism, and Goffman’s mid-20th-century work on stigma. It does so to demonstrate how in each of these contexts, social inclusion and exclusion can function as apparati that problematize people on the margins, and by extension, contribute to their governance and control. The article proposes that sociology provides a valuable orientation from which to consider social inclusion because it illuminates how social integration maintains and manages the ways in which people move about and through their socially stratified worlds.

  15. The Inclusion of Music/the Music of Inclusion

    Science.gov (United States)

    Lubet, Alex

    2009-01-01

    The intention of this paper is to situate music within inclusive education. Intersections of music--widely regarded as a "talent" or hyperability--and disability provide unique perspectives on social organisation in general and human valuation in particular. Music is a ubiquitous and an essential component of learning beginning in infancy.…

  16. Doing Research Inclusively: Bridges to Multiple Possibilities in Inclusive Research

    Science.gov (United States)

    Nind, Melanie; Vinha, Hilra

    2014-01-01

    This article reports on a study of how people do research that matters to people with learning disabilities and that involves them and their views and experiences. The study was an attempt to bring together people doing inclusive research so that, collectively, we could take stock of our practices. This would add to the individual reports and…

  17. Inclusiveness program - a SWOT analysis

    Science.gov (United States)

    Dósa, M.; Szegő, K.

    2017-09-01

    The Inclusiveness Program was created with the aim to integrate currently under-represented countries into the mainstream of European planetary research. Main stages of the working plan include setting up a database containing all the research institutes and universities where astronomical or geophysical research is carried out. It is necessary to identify their problems and needs. Challenging part of the project is to find exact means that help their work in a sustainable way. Strengths, weaknesses, opportunities and threats of the program were identified based on feedback from the inclusiveness community. Our conclusions, further suggestions are presented.

  18. Effects of kelp Ecklonia maxima inclusion in formulated feed on the ...

    African Journals Online (AJOL)

    The effect of kelp Ecklonia maxima inclusion in formulated feeds on abalone growth and gut bacterial communities has not been previously investigated in South Africa. An eight-month on-farm growth trial was conducted with sub-adult Haliotis midae (~43 mm shell length) fed graded levels of kelp in formulated feeds.

  19. [Spontaneous bacterial peritonitis].

    Science.gov (United States)

    Strauss, Edna; Caly, Wanda Regina

    2003-01-01

    Spontaneous bacterial peritonitis occurs in 30% of patients with ascites due to cirrhosis leading to high morbidity and mortality rates. The pathogenesis of spontaneous bacterial peritonitis is related to altered host defenses observed in end-stage liver disease, overgrowth of microorganisms, and bacterial translocation from the intestinal lumen to mesenteric lymph nodes. Clinical manifestations vary from severe to slight or absent, demanding analysis of the ascitic fluid. The diagnosis is confirmed by a number of neutrophils over 250/mm3 associated or not to bacterial growth in culture of an ascites sample. Enterobacteriae prevail and Escherichia coli has been the most frequent bacterium reported. Mortality rates decreased markedly in the last two decades due to early diagnosis and prompt antibiotic treatment. Third generation intravenous cephalosporins are effective in 70% to 95% of the cases. Recurrence of spontaneous bacterial peritonitis is common and can be prevented by the continuous use of oral norfloxacin. The development of bacterial resistance demands the search for new options in the prophylaxis of spontaneous bacterial peritonitis; probiotics are a promising new approach, but deserve further evaluation. Short-term antibiotic prophylaxis is recommended for patients with cirrhosis and ascites shortly after an acute episode of gastrointestinal bleeding.

  20. Social Inclusion and Integrative Practices

    Directory of Open Access Journals (Sweden)

    David Cappo

    2014-06-01

    Full Text Available With the passage of time valuable lessons have been learnt about both effective practices for program and system integration and the sizable barriers, including the challenges in sustaining constructive integration. This paper is a reflection on sustainable integrative practices and is grounded in the direct experience of one of the authors, who held the post of the South Australian Social Inclusion Commissioner. We reflect upon the structure and mechanism of the South Australian Social Inclusion Initiative (2002–2011 as well as using a case study of a successful integrative program of the Social Inclusion Initiative, a program in South Australia’s School Retention Action Plan 2004 Making the Connections (South Australian Social Inclusion Board, 2004 that was implemented to improve school retention. The case study draws out salient factors of clear rationale, coordination, collaboration, communication, team work and trust as skills and ingredients to bring about integration in policy and programs. While the integration literature affirms that these ingredients are primary skills for the development of an integrative framework, we also assert that they are not enough for successful and sustained integration. Absent from much of the literature is a discussion about the use of power and the manner in which horizontal integrative work occurs. We take up this theme to draw out some implications for analysis of sustainable integrative practices.

  1. How Bureaucracy Promotes Inclusive Organizing

    DEFF Research Database (Denmark)

    Holck, Lotte

    Diversity literature in general and Feminist in particular have long promoted alternatives to bureaucracy on the premise that this form of governance is far from gender- and race-neutral, and that inclusive organizing necessitate a flatter, decentralized and more ‘organic’ set-up (Ferguson 1984...

  2. Nonlocal quasilinear damped differential inclusions

    Directory of Open Access Journals (Sweden)

    Mouffak Benchohra

    2002-01-01

    Full Text Available In this paper we investigate the existence of mild solutions to second order initial value problems for a class of damped differential inclusions with nonlocal conditions. By using suitable fixed point theorems, we study the case when the multivalued map has convex and nonconvex values.

  3. Early Childhood Inclusion in Australia

    Science.gov (United States)

    Kemp, Coral R.

    2016-01-01

    From the introduction of early intervention services in Australian in the mid-1970s, the families of children with intellectual and multiple disabilities have been encouraged to enroll their children in local preschools and childcare centers. Children with disabilities have also accessed a range of alternatives to full inclusion, such as reverse…

  4. Teacher preparedness for inclusive education

    African Journals Online (AJOL)

    lynette

    Since the report of the National Department of Education, “Quality. Education for All” was published in 1997, it has became evident that inclusive education is going to be the way forward in special (and regular) education. Both the Consultative Paper on Special Educa- tion (30 August 1999) and the Draft White Paper on ...

  5. Inclusive Education and the Arts

    Science.gov (United States)

    Allan, Julie

    2014-01-01

    This paper addresses the troubled, problematic and contested field of inclusive education, characterised by antagonisms between so-called inclusionists and special educationists; frustration, particularly among disability activists caused by the abstraction of the social model of disability and the expansion of the special educational needs…

  6. Inclusion

    DEFF Research Database (Denmark)

    Jørgensen, Stine Kaplan

    2017-01-01

    Paperet sætter fokus på inklusion i en dans sammenhæng, med særligt fokus på mobning. Der tages udgangspunkt i et fællesskabsorienteret mobbesyn, der udpeger særlige opmærksomhedspunkter til såvel forebyggelse som intervention....

  7. Interfering with bacterial gossip

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Tolker-Nielsen, Tim; Givskov, Michael

    2011-01-01

    defense. Antibiotics exhibit a rather limited effect on biofilms. Furthermore, antibiotics have an ‘inherent obsolescence’ because they select for development of resistance. Bacterial infections with origin in bacterial biofilms have become a serious threat in developed countries. Pseudomonas aeruginosa...... that appropriately target bacteria in their relevant habitat with the aim of mitigating their destructive impact on patients. In this review we describe molecular mechanisms involved in “bacterial gossip” (more scientifically referred to as quorum sensing (QS) and c-di-GMP signaling), virulence, biofilm formation......, resistance and QS inhibition as future antimicrobial targets, in particular those that would work to minimize selection pressures for the development of resistant bacteria....

  8. INCLUSIVE CULTURE IN PRE-SCHOOL INSTITUTIONS

    Directory of Open Access Journals (Sweden)

    Irena NOVACHEVSKA

    2011-04-01

    Full Text Available Inclusive education is a rational concept that refers to the overall and long-term transformation of institutional systems in society, especially in education. Along with the transformation, a number of important and unresolved issues still appear in both theory and practice, as the duty of pre-school institutions and schools is to educate every student in the mainstream education system. One of the most important aspects of inclusion is the inclusive culture. Regardless of the good inclusive policy and practice, one cannot talk about successful inclusion without a properly developed inclusive institutional culture.This paper is a contribution to the research considering the development of inclusive culture in three preschool institutions. It is based on the thinking and attitudes of the pre­school staff toward the necessity of developing and nurturing an inclusive culture. Successful inclusion of pupils with special needs in the mainstream school system cannot be conceived without an inclusive culture.

  9. Primordial Compositions of Refractory Inclusions

    Energy Technology Data Exchange (ETDEWEB)

    Grossman, L; Simon, S B; Rai, V K; Thiemens, M H; Hutcheon, I D; Williams, R W; Galy, A; Ding, T; Fedkin, A V; Clayton, R N; Mayeda, T K

    2008-02-20

    Bulk chemical and oxygen, magnesium and silicon isotopic compositions were measured for each of 17 Types A and B refractory inclusions from CV3 chondrites. After bulk chemical compositions were corrected for non-representative sampling in the laboratory, the Mg and Si isotopic compositions of each inclusion were used to calculate its original chemical composition assuming that the heavy-isotope enrichments of these elements are due to Rayleigh fractionation that accompanied their evaporation from CMAS liquids. The resulting pre-evaporation chemical compositions are consistent with those predicted by equilibrium thermodynamic calculations for high-temperature nebular condensates but only if different inclusions condensed from nebular regions that ranged in total pressure from 10{sup -6} to 10{sup -1} bar, regardless of whether they formed in a system of solar composition or in one enriched in OC dust relative to gas by a factor of ten relative to solar composition. This is similar to the range of total pressures predicted by dynamic models of the solar nebula for regions whose temperatures are in the range of silicate condensation temperatures. Alternatively, if departure from equilibrium condensation and/or non-representative sampling of condensates in the nebula occurred, the inferred range of total pressure could be smaller. Simple kinetic modeling of evaporation successfully reproduces observed chemical compositions of most inclusions from their inferred pre-evaporation compositions, suggesting that closed-system isotopic exchange processes did not have a significant effect on their isotopic compositions. Comparison of pre-evaporation compositions with observed ones indicates that 80% of the enrichment in refractory CaO + Al{sub 2}O{sub 3} relative to more volatile MgO + SiO{sub 2} is due to initial condensation and 20% due to subsequent evaporation for both Type A and Type B inclusions.

  10. Foreign Body

    Science.gov (United States)

    ... SearchingPediatrics.com Pediatrics Common Questions, Quick Answers Foreign Body Donna D'Alessandro, M.D. Lindsay Huth, B. ... I call the doctor? What is a foreign body? A foreign body is when an object is ...

  11. Inclusive teaching strategies - dimensions and agendas

    DEFF Research Database (Denmark)

    Molbæk, Mette

    2017-01-01

    will contribute to an enrichment of the two research fields and give teachers and researchers new opportunities for developing more inclusive schools. Based on research focused on teaching strategies that support students’ opportunities for learning, four dimensions of inclusive teaching strategies are presented...... those teachers working with inclusion can be supported with more tangible and meaningful ways of understanding and developing inclusive classrooms. This article argues that seeing research in inclusion in close connection with research in effective teaching strategies and classroom management...

  12. Inclusion control in high-performance steels

    International Nuclear Information System (INIS)

    Holappa, L.E.K.; Helle, A.S.

    1995-01-01

    Progress of clean steel production, fundamentals of oxide and sulphide inclusions as well as inclusion morphology in normal and calcium treated steels are described. Effects of cleanliness and inclusion control on steel properties are discussed. In many damaging constructional and engineering applications the nonmetallic inclusions have a quite decisive role in steel performance. An example of combination of good mechanical properties and superior machinability by applying inclusion control is presented. (author)

  13. Bacterial surface adaptation

    Science.gov (United States)

    Utada, Andrew

    2014-03-01

    Biofilms are structured multi-cellular communities that are fundamental to the biology and ecology of bacteria. Parasitic bacterial biofilms can cause lethal infections and biofouling, but commensal bacterial biofilms, such as those found in the gut, can break down otherwise indigestible plant polysaccharides and allow us to enjoy vegetables. The first step in biofilm formation, adaptation to life on a surface, requires a working knowledge of low Reynolds number fluid physics, and the coordination of biochemical signaling, polysaccharide production, and molecular motility motors. These crucial early stages of biofilm formation are at present poorly understood. By adapting methods from soft matter physics, we dissect bacterial social behavior at the single cell level for several prototypical bacterial species, including Pseudomonas aeruginosa and Vibrio cholerae.

  14. Bacterial Meningitis in Infants

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2004-04-01

    Full Text Available A retrospective study of 80 infantile patients (ages 30-365 days; 47 male, 33 female with culture-proven bacterial meningitis seen over a 16 year period (1986-2001 is reported from Taiwan.

  15. Factitious Bacterial Meningitis Revisited

    Science.gov (United States)

    Peterson, E.; Thrupp, L.; Uchiyama, N.; Hawkins, B.; Wolvin, B.; Greene, G.

    1982-01-01

    Nonviable gram-negative bacilli were seen in smears of cerebrospinal fluid from eight infants in whom bacterial meningitis was ruled out. Tubes from commercial kits were the source of the factitious organisms. PMID:7153328

  16. Vimentin in Bacterial Infections

    DEFF Research Database (Denmark)

    Mak, Tim N; Brüggemann, Holger

    2016-01-01

    filaments (IFs). IFs have not only roles in maintaining the structural integrity of the cell, but they are also involved in many cellular processes including cell adhesion, immune signaling, and autophagy, processes that are important in the context of bacterial infections. Here, we summarize the knowledge...... about the role of IFs in bacterial infections, focusing on the type III IF protein vimentin. Recent studies have revealed the involvement of vimentin in host cell defenses, acting as ligand for several pattern recognition receptors of the innate immune system. Two main aspects of bacteria......-vimentin interactions are presented in this review: the role of vimentin in pathogen-binding on the cell surface and subsequent bacterial invasion and the interaction of cytosolic vimentin and intracellular pathogens with regards to innate immune signaling. Mechanistic insight is presented involving distinct bacterial...

  17. [Diagnosis of bacterial vaginosis].

    Science.gov (United States)

    Djukić, Slobodanka; Ćirković, Ivana; Arsić, Biljana; Garalejić, Eliana

    2013-01-01

    Bacterial vaginosis is a common, complex clinical syndrome characterized by alterations in the normal vaginal flora. When symptomatic, it is associated with a malodorous vaginal discharge and on occasion vaginal burning or itching. Under normal conditions, lactobacilli constitute 95% of the bacteria in the vagina. Bacterial vaginosis is associated with severe reduction or absence of the normal H2O2-producing lactobacilli and overgrowth of anaerobic bacteria and Gardnerella vaginalis, Atopobium vaginae, Mycoplasma hominis and Mobiluncus species. Most types of infectious disease are diagnosed by culture, by isolating an antigen or RNA/DNA from the microbe, or by serodiagnosis to determine the presence of antibodies to the microbe. Therefore, demonstration of the presence of an infectious agent is often a necessary criterion for the diagnosis of the disease. This is not the case for bacterial vaginosis, since the ultimate cause of the disease is not yet known. There are a variety of methods for the diagnosis of bacterial vaginosis but no method can at present be regarded as the best. Diagnosing bacterial vaginosis has long been based on the clinical criteria of Amsel, whereby three of four defined criteria must be satisfied. Nugent's scoring system has been further developed and includes validation of the categories of observable bacteria structures. Up-to-date molecular tests are introduced, and better understanding of vaginal microbiome, a clear definition for bacterial vaginosis, and short-term and long-term fluctuations in vaginal microflora will help to better define molecular tests within the broader clinical context.

  18. The clinical impact of bacterial biofilms

    DEFF Research Database (Denmark)

    Høiby, Niels; Ciofu, Oana; Johansen, Helle Krogh

    2011-01-01

    Bacteria survive in nature by forming biofilms on surfaces and probably most, if not all, bacteria (and fungi) are capable of forming biofilms. A biofilm is a structured consortium of bacteria embedded in a self-produced polymer matrix consisting of polysaccharide, protein and extracellular DNA....... Bacterial biofilms are resistant to antibiotics, disinfectant chemicals and to phagocytosis and other components of the innate and adaptive inflammatory defense system of the body. It is known, for example, that persistence of staphylococcal infections related to foreign bodies is due to biofilm formation....... Likewise, chronic Pseudomonas aeruginosa lung infections in cystic fibrosis patients are caused by biofilm growing mucoid strains. Gradients of nutrients and oxygen exist from the top to the bottom of biofilms and the bacterial cells located in nutrient poor areas have decreased metabolic activity...

  19. STUDENTS’ RESPONSES TO INCLUSIVE DESIGN

    DEFF Research Database (Denmark)

    Herriott, Richard; Geert Jensen, Birgitte

    2012-01-01

    . This technology is designed help to citizens to be 'master of his or her own life' while off-setting capability changes due declining physical ability. Welfare technology includes smarter working practices or service concepts, which frees up labor resources and it also covers robotics, telemedicine, IT solutions...... and intelligent devices. This paper will show the results of students' work with the problem of changed demographics and emerging needs in products and services. In so doing it looks at how the use of inclusive design methods affects students´work processes. This is done by the use of work diaries which provided......This paper looks at how students’ design process responds to the requirements of inclusive design. The background to the students´ brief was the concept of welfare technology. People wish to retain their customary life-style even as ageing brings with it a reduction in physical capability: loss...

  20. The HLT inclusive B triggers

    CERN Document Server

    Gligorov, Vladimir V; Williams, Michael

    2011-01-01

    The inclusive HLT strategy relies on triggering any B decay based on two signatures: a single significantly displaced, high transverse momentum track, and a significantly displaced vertex containing this track and 1-3 other tracks, with high total transverse momentum. In order to provide optimal signal efficiency and background rejection the displaced vertex selection is implemented in a novel boosted decision tree algorithm incorporating information about the experimental resolution in the boosting procedure to protect against overtraining. The performance of these triggers has been commissioned using data taken during 2011 LHCb running and is evaluated here in a data-driven manner. The HLT inclusive triggers are found to have a rejection factor of around 1000 with respect to events selected by the L0 hardware trigger and a bbar purity close to 100%.

  1. Interprofessional development in inclusive schools

    DEFF Research Database (Denmark)

    Hansen, Britt Blaabjerg; Højholdt, Andy; Arndal, Lars Stubbe

    circles (Persson, 2007). The design development was theory oriented, with our introducing theories about the different aspects of professional collaboration, co-creation and social innovation (Conger, S. 2002, Edwall et. al. 2008)).In accordance with the processes of design-based research (iteration...... required to work with these students. This paper describes the challenges [education/school] professionals (teachers and child and youth educators) face in striving to develop inclusive school practice, and how these challenges relate to differences in skills and competencies between the professions....... The knowledge for the paper was generated in a research project entitled “Interprofessional development in inclusive schools”. The project worked from a basic understanding that we need to develop preventive educational strategies for children and young people with learning difficulties and in social risk...

  2. Friendship in inclusive physical education.

    Science.gov (United States)

    Seymour, Helena; Reid, Greg; Bloom, Gordon A

    2009-07-01

    Social interaction and development of friendships between children with and without a disability are often proposed as potential outcomes of inclusive education. Physical activity specialists assert that exercise and sport environments may be conducive to social and friendship outcomes. This study investigated friendship in inclusive physical education from the perspective of students with (n = 8) and without (n = 8) physical disabilities. All participants attended a reversely integrated school and were interviewed using a semistructured, open-ended format. An adapted version of Weiss, Smith, and Theeboom's (1996) interview guide exploring perceptions of peer relationships in the sport domain was used. Four conceptual categories emerged from the analysis: development of friendship, best friend, preferred physical activities and outcomes, and dealing with disability. The results demonstrated the key characteristics of best friends and the influential role they play.

  3. Social inclusion and integrative practices

    OpenAIRE

    Cappo, David; Verity, Fiona

    2014-01-01

    With the passage of time valuable lessons have been learnt about both effective practices for program and system integration and the sizable barriers, including the challenges in sustaining constructive integration. This paper is a reflection on sustainable integrative practices and is grounded in the direct experience of one of the authors, who held the post of the South Australian Social Inclusion Commissioner. We reflect upon the structure and mechanism of the South Australian Social Inclu...

  4. Student' responses to inclusive design

    DEFF Research Database (Denmark)

    Herriott, Richard; Geert Jensen, Birgitte

    2013-01-01

    ” and “disordered”, this study visualises it and finds patterns underlying a process which is perceived to be chaotic. It provides a rough measure of the dynamics of a project in the form of the “switch between” ratio: how many changes in activity take place from the start to the finish of a project. The article...... was originally written for the DRTS 2012 conference "Articulating Design Thinking" and proposed for inclusion in a special edition of Design Issues....

  5. The Sociology of Social Inclusion

    OpenAIRE

    Dan Allman

    2013-01-01

    This article looks at social inclusion from a sociological perspective. It argues that sociology complements biological and other natural order explanations of social stratification. The article interrogates a variety of forms of social integration, including ostracism within 5th century b.c. Greece, 19th-century solidarism, and Goffman’s mid-20th-century work on stigma. It does so to demonstrate how in each of these c...

  6. Student Engagement In Inclusive Classrooms

    DEFF Research Database (Denmark)

    Rangvid, Beatrice Schindler

    There is general agreement that to thrive and learn at their best, students must be engaged. However, schools face a particular challenge to provide a suitable and engaging learning environment for SEN (special educational needs) students who are educated in general education classes. Using data......-students as for other students. This highlights the need for better inclusion initiatives aimed at strengthening engagement of SEN-students in regular classrooms....

  7. Inclusive fitness theory and eusociality

    DEFF Research Database (Denmark)

    Abbot, Patrick; Abe, Jun; Alcock, John

    2011-01-01

    Arising from M. A. Nowak, C. E. Tarnita & E. O. Wilson 466, 1057-1062 (2010); Nowak et al. reply. Nowak et al. argue that inclusive fitness theory has been of little value in explaining the natural world, and that it has led to negligible progress in explaining the evolution of eusociality. However......, we believe that their arguments are based upon a misunderstanding of evolutionary theory and a misrepresentation of the empirical literature. We will focus our comments on three general issues....

  8. Does financial inclusion affect monetary policy in SAARC countries?

    Directory of Open Access Journals (Sweden)

    Sanjaya Kumar Lenka

    2016-12-01

    Full Text Available Alike the role of heart for human body, finance is the focal point of an economy, whereas savings and investment are its tubes and vessels. Hence, a solid financial system is a fundamental character of an enduring economy. The frozen financial system endures longer if its foundation is concrete and subsists in the people of grass-root level. They are those, who live in villages and small towns, earn meager income, work in primary sector, spend more on food, and have lesser social securities. In this setting, the process of bringing these people into the main stream of financial activities is called financial inclusion. This study describes the impact of financial inclusion on monetary policy of South Asian Association for Regional Cooperation (SAARC countries from 2004–2013. The study uses principal component analysis (PCA to construct a Financial Inclusion Index that serves as a proxy variable for the accessibility of financial inclusion in the SAARC countries. Adding to it, three different models like FEM, REM, and Panel-corrected standard errors are used for the analysis. In this study, an empirical result of generalized least square(GLS estimation shows that financial inclusion, exchange rate, and interest rate are negatively associated with inflation in SAARC countries.

  9. Perceived and actual social discrimination: the case of overweight and social inclusion.

    Science.gov (United States)

    Hartung, Freda-Marie; Renner, Britta

    2013-01-01

    The present study examined the correspondence between perceived and actual social discrimination of overweight people. In total, 77 first-year students provided self-ratings about their height, weight, and perceived social inclusion. To capture actual social inclusion, each participant nominated those fellow students (a) she/he likes and dislikes and (b) about whom she/he is likely to hear social news. Students with lower Body Mass Index (BMI) felt socially included, irrespective of their actual social inclusion. In contrast, students with higher BMI felt socially included depending on the degree of their actual social inclusion. Specifically, their felt social inclusion accurately reflected whether they were actually liked/disliked, but only when they were part of social news. When not part of social news, they also showed insensitivity to their actual social inclusion status. Thus, students with a lower BMI tended to be insensitive, while students with a higher BMI showed a differential sensitivity to actual social discrimination.

  10. Inclusive and semi-inclusive hadron interactions at ISR energies

    International Nuclear Information System (INIS)

    Giacomelli, G.

    1987-01-01

    Experiments at the CERN proton-antiproton collider and at the ISR have recently provided a wealth of information on high-energy, low-transverse momentum hadron-hadron interactions. These data are changing the picture of inelastic collisions at high-energies. In this paper will be mainly discussed high-statistics data from the ABCDHW collaboration using the Split Field Magnet (SFM) detector at the CERN-ISR. In particular will be discussed inclusive distributions as functions of x and y, the multiplicity and rapidity dependence of the mean transverse momentum, its fluctuations, correlations in rapidity, Bose-Einstein correlations, charged multiplicity distributions and rapidity fluctuations

  11. Bacterial Cell Mechanics.

    Science.gov (United States)

    Auer, George K; Weibel, Douglas B

    2017-07-25

    Cellular mechanical properties play an integral role in bacterial survival and adaptation. Historically, the bacterial cell wall and, in particular, the layer of polymeric material called the peptidoglycan were the elements to which cell mechanics could be primarily attributed. Disrupting the biochemical machinery that assembles the peptidoglycan (e.g., using the β-lactam family of antibiotics) alters the structure of this material, leads to mechanical defects, and results in cell lysis. Decades after the discovery of peptidoglycan-synthesizing enzymes, the mechanisms that underlie their positioning and regulation are still not entirely understood. In addition, recent evidence suggests a diverse group of other biochemical elements influence bacterial cell mechanics, may be regulated by new cellular mechanisms, and may be triggered in different environmental contexts to enable cell adaptation and survival. This review summarizes the contributions that different biomolecular components of the cell wall (e.g., lipopolysaccharides, wall and lipoteichoic acids, lipid bilayers, peptidoglycan, and proteins) make to Gram-negative and Gram-positive bacterial cell mechanics. We discuss the contribution of individual proteins and macromolecular complexes in cell mechanics and the tools that make it possible to quantitatively decipher the biochemical machinery that contributes to bacterial cell mechanics. Advances in this area may provide insight into new biology and influence the development of antibacterial chemotherapies.

  12. Specific chlamydial inclusion membrane proteins associate with active Src family kinases in microdomains that interact with the host microtubule network.

    Science.gov (United States)

    Mital, Jeffrey; Miller, Natalie J; Fischer, Elizabeth R; Hackstadt, Ted

    2010-09-01

    Chlamydiae are Gram-negative obligate intracellular bacteria that cause diseases with significant medical and economic impact. Chlamydia trachomatis replicates within a vacuole termed an inclusion, which is extensively modified by the insertion of a number of bacterial effector proteins known as inclusion membrane proteins (Incs). Once modified, the inclusion is trafficked in a dynein-dependent manner to the microtubule-organizing centre (MTOC), where it associates with host centrosomes. Here we describe a novel structure on the inclusion membrane comprised of both host and bacterial proteins. Members of the Src family of kinases are recruited to the chlamydial inclusion in an active form. These kinases display a distinct, localized punctate microdomain-like staining pattern on the inclusion membrane that colocalizes with four chlamydial inclusion membrane proteins (Incs) and is enriched in cholesterol. Biochemical studies show that at least two of these Incs stably interact with one another. Furthermore, host centrosomes associate with these microdomain proteins in C. trachomatis-infected cells and in uninfected cells exogenously expressing one of the chlamydial effectors. Together, the data suggest that a specific structure on the C. trachomatis inclusion membrane may be responsible for the known interactions of chlamydiae with the microtubule network and resultant effects on centrosome stability.

  13. Event by Event fluctuations and Inclusive Distribution

    OpenAIRE

    Bialas, A.; Koch, V.

    1999-01-01

    Event-by-event observables are compared with conventional inclusive measurements. We find that moments of event-by-event fluctuations are closely related to inclusive correlation functions. Implications for upcomming heavy ion experiments are discussed.

  14. Bacterial meningitis in children

    International Nuclear Information System (INIS)

    Marji, S.

    2007-01-01

    To demonstrate the epidemiology, clinical manifestations and bacteriological profile of bacterial meningitis in children beyond the neonatal period in our hospital. This was a retrospective descriptive study conducted at Prince Rashid Hospital in Irbid, Jordan. The medical records of 50 children with the diagnosis of bacterial meningitis during 4 years period, were reviewed. The main cause of infection was streptococcus pneumoniae, followed by Haemophilus influenza and Niesseria meningitides. Mortality was higher in infants and meningococcal infection, while complications were more encountered in cases of streptococcus pneumoniae. Cerebrospinal fluid culture was positive in 11 cases and Latex agglutination test in 39. There is a significant reduction of the numbers of bacterial meningitis caused by Haemophilus influenza type B species. (author)

  15. Interfering with bacterial gossip

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Tolker-Nielsen, Tim; Givskov, Michael

    2011-01-01

    Biofilm resilience poses major challenges to the development of novel antimicrobial agents. Biofilm bacteria can be considered small groups of “Special Forces” capable of infiltrating the host and destroying important components of the cellular defense system with the aim of crippling the host...... defense. Antibiotics exhibit a rather limited effect on biofilms. Furthermore, antibiotics have an ‘inherent obsolescence’ because they select for development of resistance. Bacterial infections with origin in bacterial biofilms have become a serious threat in developed countries. Pseudomonas aeruginosa...... that appropriately target bacteria in their relevant habitat with the aim of mitigating their destructive impact on patients. In this review we describe molecular mechanisms involved in “bacterial gossip” (more scientifically referred to as quorum sensing (QS) and c-di-GMP signaling), virulence, biofilm formation...

  16. Inclusive outreach practices in Palaeontology: Inclusive-Coworking

    Science.gov (United States)

    García-Frank, Alejandra; Gomez-Heras, Miguel; Fesharaki, Omid

    2017-04-01

    Previous experiences with people with both physical and intellectual functional diversity around palaeontological issues have demonstrated the important value of science outreach directed to people with disabilities. The aforementioned practices act twofold: as a learning tool and also improving the quality of life of the participants and thus, their self-image. All these pioneer experiences were the first step in a process of developing new attitudes contributing the 2030 Agenda for Sustainable Development of United Nations, where among the 17 goals proposed an effective social inclusion of people with disabilities is required. For this, real inclusive practices in geological outreach are imperious. A close cooperation with all the parts (researchers and participants), in a kind of coworking attitude is needed. This Inclusive-Coworking is considered in the sense of social gathering in order to share equal values and look for the synergy that this different outlook implies. And what is more important: the change of role of the previously learners into an active part of the scientific outreach, providing the adequate methodology for that. The offer of non-formal learning activities normally includes the participation of university professors and researchers in Science Week editions. During the 2016 session in Madrid, four adults with intellectual disability who were participants in the previous edition, contributed in the palaeontological workshop. They were in charge of four of the eight modules explaining the origin of fossils and how to collect them, the evolution of equids' limbs, and the main dentition types in vertebrates to the twenty 16 year old secondary students who attended the workshop. During the development of the experience all the students were pleased with the inclusive approach, and the interaction of all participants was fruitful. Although the explanations took a bit more time when made by our functional diverse fellows, all the abstracts concepts

  17. Diagnosis of bacterial vaginosis

    Directory of Open Access Journals (Sweden)

    Đukić Slobodanka

    2013-01-01

    Full Text Available Bacterial vaginosis is a common, complex clinical syndrome characterized by alterations in the normal vaginal flora. When symptomatic, it is associated with a malodorous vaginal discharge and on occasion vaginal burning or itching. Under normal conditions, lactobacilli constitute 95% of the bacteria in the vagina. Bacterial vaginosis is associated with severe reduction or absence of the normal H2O2­producing lactobacilli and overgrowth of anaerobic bacteria and Gardnerella vaginalis, Atopobium vaginae, Mycoplasma hominis and Mobiluncus species. Most types of infectious disease are diagnosed by culture, by isolating an antigen or RNA/DNA from the microbe, or by serodiagnosis to determine the presence of antibodies to the microbe. Therefore, demonstration of the presence of an infectious agent is often a necessary criterion for the diagnosis of the disease. This is not the case for bacterial vaginosis, since the ultimate cause of the disease is not yet known. There are a variety of methods for the diagnosis of bacterial vaginosis but no method can at present be regarded as the best. Diagnosing bacterial vaginosis has long been based on the clinical criteria of Amsel, whereby three of four defined criteria must be satisfied. Nugent’s scoring system has been further developed and includes validation of the categories of observable bacteria structures. Up­to­date molecular tests are introduced, and better understanding of vaginal microbiome, a clear definition for bacterial vaginosis, and short­term and long­term fluctuations in vaginal microflora will help to better define molecular tests within the broader clinical context.

  18. Adult bacterial meningitis

    DEFF Research Database (Denmark)

    Meyer, C N; Samuelsson, I S; Galle, M

    2004-01-01

    Episodes of adult bacterial meningitis (ABM) at a Danish hospital in 1991-2000 were identified from the databases of the Department of Clinical Microbiology, and compared with data from the Danish National Patient Register and the Danish National Notification System. Reduced penicillin susceptibi......Episodes of adult bacterial meningitis (ABM) at a Danish hospital in 1991-2000 were identified from the databases of the Department of Clinical Microbiology, and compared with data from the Danish National Patient Register and the Danish National Notification System. Reduced penicillin...

  19. Bacterial blight of cotton

    Directory of Open Access Journals (Sweden)

    Aïda JALLOUL

    2015-04-01

    Full Text Available Bacterial blight of cotton (Gossypium ssp., caused by Xanthomonas citri pathovar malvacearum, is a severe disease occurring in all cotton-growing areas. The interactions between host plants and the bacteria are based on the gene-for-gene concept, representing a complex resistance gene/avr gene system. In light of the recent data, this review focuses on the understanding of these interactions with emphasis on (1 the genetic basis for plant resistance and bacterial virulence, (2 physiological mechanisms involved in the hypersensitive response to the pathogen, including hormonal signaling, the oxylipin pathway, synthesis of antimicrobial molecules and alteration of host cell structures, and (3 control of the disease.

  20. Bacterial meningitis in infants.

    Science.gov (United States)

    Ku, Lawrence C; Boggess, Kim A; Cohen-Wolkowiez, Michael

    2015-03-01

    Neonatal bacterial meningitis is uncommon but devastating. Morbidity among survivors remains high. The types and distribution of pathogens are related to gestational age, postnatal age, and geographic region. Confirming the diagnosis is difficult. Clinical signs are often subtle, lumbar punctures are frequently deferred, and cerebrospinal fluid (CSF) cultures can be compromised by prior antibiotic exposure. Infants with bacterial meningitis can have negative blood cultures and normal CSF parameters. Promising tests such as the polymerase chain reaction require further study. Prompt treatment with antibiotics is essential. Clinical trials investigating a vaccine for preventing neonatal Group B Streptococcus infections are ongoing. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Regge cuts in inclusive reactions

    International Nuclear Information System (INIS)

    Paige, F.E.; Trueman, T.L.

    1975-01-01

    The contribution of Regge cuts to single-particle inclusive processes is analyzed using the techniques of Gribov. The dependence of these contributions on the polarization state of the target is emphasized. A general formula is obtained and certain contributions to it are calculated. It is not possible, however, to reduce this to a simple, powerful formula expressing the total cut contribution in terms of other measurable quantities, as can be done for the cut contribution to the total cross section. The reasons for this are discussed in detail. The single-particle intermediate states, analogous to the absorption model for elastic scattering, are explicitly calculated as an illustration

  2. Teaching Competences and Inclusive Education

    Directory of Open Access Journals (Sweden)

    José María Fernández Batanero

    2013-08-01

    Full Text Available This paper focuses on teaching competencies that are conducive to good educational practices in relation to inclusion, from the perspective of teachers. The methodology employed in the study is descriptive/comprehensive, and of an exploratory nature. By means of four case studies, the perceptions of teachers from two secondary schools—characterized by the Spanish Educational Administration as having “good practices”— are examined. The techniques used for information collection in this study include documentary analysis, in-depth interviews and focus groups. The findings emphasize the importance of strategic skills, combined with innovation and creativity, among others.

  3. Evaluation of a Bacillus direct-fed microbial candidate on digesta viscosity, bacterial translocation, microbiota composition and bone mineralisation in broiler chickens fed on a rye-based diet.

    Science.gov (United States)

    Latorre, J D; Hernandez-Velasco, X; Bielke, L R; Vicente, J L; Wolfenden, R; Menconi, A; Hargis, B M; Tellez, G

    2015-01-01

    1. The effects of the dietary inclusion of a Bacillus-based direct-fed microbial (DFM) candidate on digesta viscosity, bacterial translocation, microbiota composition and bone mineralisation were evaluated in broilers consuming rye-based diets. 2. In the present study, control mash rye-based diets (CON) or Bacillus-DFM supplemented diets (TRT) were administered ad libitum to male broilers in three independent experiments. 3. In Experiments 1 and 2 (n = 25/group), liver samples were taken to evaluate bacterial translocation, digesta samples were used for viscosity measurements and the intestinal microbial flora was evaluated from different intestinal sections to enumerate total recovered gram-negative bacteria (TGB), lactic acid bacteria (LAB) and anaerobic bacteria (TAB). Additionally, both tibias were removed for assessment of bone quality. 4. In Experiment 3, each experimental group had 8 replicates of 20 chickens (n = 160/group). Weekly, body weight (BW), feed intake (FI) and feed conversion ratio (FCR) were evaluated. At d 28-of-age, samples were taken to determine bacterial translocation, digesta viscosity and bone quality characteristics. 5. In all experiments, consumption of Bacillus-DFM reduced bacterial translocation to the liver and digesta viscosity. Additionally, DFM supplementation improved BW, bone quality measurements and FCR. Moreover, chickens fed on the Bacillus-DFM diet in Experiments 1 and 2 showed a significant reduction in the number of gram-negative and anaerobic bacteria in the duodenal content compared to control. 6. In summary, chickens fed on a rye-based diet without DFM inclusion showed an increase in bacterial translocation and digesta viscosity, accompanied by reduced performance and bone quality variables relative to the Bacillus-DFM candidate group. Hence, incorporation into the feed of a selected DFM ameliorated the adverse anti-nutritional effects related to utilisation of rye-based diets in broilers chickens.

  4. Principals' Knowledge of and Attitudes toward Inclusion.

    Science.gov (United States)

    Barnett, Carol; Monda-Amaya, Lisa E.

    1998-01-01

    A survey of 65 elementary, junior, and high school principals investigated attitudes toward and knowledge of inclusion. No clear definition of inclusion emerged, but principals generally viewed inclusion as most appropriate for students with mild disabilities. Also, results indicated that teachers were not adequately prepared to implement…

  5. Inclusive Education: Examining Equity on Five Continents

    Science.gov (United States)

    Artiles, Alfredo J., Ed.; Kozleski, Elizabeth B., Ed.; Waitoller, Federico R., Ed.

    2011-01-01

    Despite the impressive growth of inclusive education around the world, questions and considerations about equity have been neglected. This edited volume makes a major contribution to the field of inclusive education by analyzing equity concerns that have emerged from the implementation of inclusive education models in nine nations on five…

  6. 75 FR 81395 - Minority and Women Inclusion

    Science.gov (United States)

    2010-12-28

    ... 2590-AA28 Minority and Women Inclusion AGENCIES: Federal Housing Finance Board; Federal Housing Finance... and the inclusion of women and minorities in all activities. The final rule implements the provisions.... It also requires each regulated entity to establish an Office of Minority and Women Inclusion, or...

  7. 75 FR 10446 - Minority and Women Inclusion

    Science.gov (United States)

    2010-03-08

    ... 2590-AA28 Minority and Women Inclusion AGENCIES: Federal Housing Finance Board; Federal Housing Finance... minority and women inclusion. Section 1116 of the Housing and Economic Recovery Act of 2008 amended section... Loan Banks to promote diversity and the inclusion of women and minorities in all activities...

  8. Teachers' Experience with Inclusive Education in Singapore

    Science.gov (United States)

    Yeo, Lay See; Chong, Wan Har; Neihart, Maureen F.; Huan, Vivien S.

    2016-01-01

    Teachers' positive attitude is most critically and consistently associated with successful inclusion. However, little is known about teachers' first-hand encounters with inclusive education in Singapore. We present findings from a qualitative study on inclusion based on focus group interviews with 202 teachers from 41 resourced primary schools.…

  9. "Inclusion in Practice": Does Practice Make Perfect?

    Science.gov (United States)

    Slee, Roger

    2001-01-01

    Discusses four issues raised in this special issue: (1) problems with the term "inclusion"; (2) reductionist approaches to inclusive education; (3) politics of special education research and the need to include parent and student voices; and (4) preparation of teachers for inclusive education. (Contains 53 references.) (SK)

  10. Inclusive Classes in Physical Education: Teachers' Difficulties

    Science.gov (United States)

    Toloi, Gabriela Gallucci; Manzini, Eduardo José; Spoldaro, Diego Machado; Zacarias, Lucas Ventura

    2016-01-01

    The successful inclusion of students with special needs in physical education classes requires much planning and preparation. Lack of preparation of physical education teachers working in inclusive settings in Brazil has demonstrated the need for specialized training in strategies for implementing inclusion. The goal of this study was to identify,…

  11. Ultrastructural cytochemical analysis of intranuclear arsenic inclusions

    Energy Technology Data Exchange (ETDEWEB)

    Sorensen, E.M.B.

    1987-01-01

    To establish the chemical composition of the arsenic inclusion, freshly isolated preparations of inclusions and epon-embedded thin sections of inclusions were subjected to ultrastructural cytochemical analysis. Intranuclear inclusions are composed of amorphous, arsenic-containing subunits aligned linearly to form a coiled complex. Lipase, ribonuclease, deoxyribonuclease, trypsin, pepsin, protease, amylase, or ethylenediaminetetraacetic acid (EDTA) was used to digest or chelate these inclusions. Following enzymatic digestion or chelation, the electron opacity of inclusions was compared with that of control sections exposed for equal times to equivalent solutions lacking the enzymes. Exposure to amylase caused a consistent reduction in the electron opacity of thin sections of inclusions and almost complete digestion of the freshly isolated preparations of inclusions. This was indicative of the presence of a carbohydrate moiety within arsenic inclusions. Incubation of inclusions with EDTA resulted in solubilization of freshly isolated and thin-sectioned embedded material. These data indicated that the intranuclear arsenic inclusion is composed of both metallic and carbohydrate moieties, confirming earlier studies which identified arsenic within inclusions using instrumental neutron activation analysis and x-ray microprobe analysis.

  12. Full averaging of fuzzy impulsive differential inclusions

    Directory of Open Access Journals (Sweden)

    Natalia V. Skripnik

    2010-09-01

    Full Text Available In this paper the substantiation of the method of full averaging for fuzzy impulsive differential inclusions is studied. We extend the similar results for impulsive differential inclusions with Hukuhara derivative (Skripnik, 2007, for fuzzy impulsive differential equations (Plotnikov and Skripnik, 2009, and for fuzzy differential inclusions (Skripnik, 2009.

  13. Elements of Inclusion: Findings from the Field

    Science.gov (United States)

    McMaster, Christopher

    2014-01-01

    The Ministry of Education has set the target of 100% of New Zealand schools to be "mostly" inclusive by 2014. But what are the essential elements of inclusion? This paper explores essential core elements that allow inclusion to flourish. Based on an extensive time in the field as part of a year-long doctoral research project, these…

  14. Curcumin-Hydroxypropyl-β-Cyclodextrin Inclusion Complex ...

    African Journals Online (AJOL)

    Purpose: To evaluate the effect of the preparation method on the inclusion complex of curcumin and hydroxypropyl-β-cyclodextrin Methods: HP-β-CD was selected to prepare an inclusion complex with curcumin at a molar ratio of 1:1. The inclusion complexes were prepared using three different methods: common solvent ...

  15. Measuring Inclusive Education Outcomes in Alberta, Canada

    Science.gov (United States)

    Loreman, Tim

    2014-01-01

    This study details the results of a review of the academic and public sector literature on measuring inclusive education in large systems. It highlights some outcomes drawn from the international literature on inclusion that might be indicative of the presence and quality of inclusive education in an effort to develop a set of outcomes for…

  16. Existence of solutions of functional differential inclusions

    Directory of Open Access Journals (Sweden)

    A. Anguraj

    1992-01-01

    Full Text Available We prove the existence of solutions of a functional differential inclusion. By using the variation of parameters formula we convert the functional differential inclusion into an integral inclusion and prove the existence of a fixed point of the set-valued mapping with the help of the Kakutani-Bohnenblust-Karlin fixed point theorem.

  17. Body Image

    Science.gov (United States)

    ... affect body image Pre-baby body Pregnancy and eating disorders Looking for information on mental health conditions? Visit ... Mental health section. Fact sheets Anorexia nervosa Binge eating disorder Bulimia nervosa Cosmetics and your health Depression during ...

  18. Bacterial Metabolism Shapes the Host-Pathogen Interface.

    Science.gov (United States)

    Passalacqua, Karla D; Charbonneau, Marie-Eve; O'Riordan, Mary X D

    2016-06-01

    Bacterial pathogens have evolved to exploit humans as a rich source of nutrients to support survival and replication. The pathways of bacterial metabolism that permit successful colonization are surprisingly varied and highlight remarkable metabolic flexibility. The constraints and immune pressures of distinct niches within the human body set the stage for understanding the mechanisms by which bacteria acquire critical nutrients. In this article we discuss how different bacterial pathogens carry out carbon and energy metabolism in the host and how they obtain or use key nutrients for replication and immune evasion.

  19. Bacterial metabolism shapes the host:pathogen interface

    Science.gov (United States)

    Passalacqua, Karla D.; Charbonneau, Marie-Eve; O'Riordan, Mary X.D.

    2016-01-01

    Summary Bacterial pathogens have evolved to exploit humans as a rich source of nutrients to support survival and replication. The pathways of bacterial metabolism that permit successful colonization are surprisingly varied and highlight remarkable metabolic flexibility. The constraints and immune pressures of distinct niches within the human body set the stage for understanding the mechanisms by which bacteria acquire critical nutrients. Here we discuss how different bacterial pathogens carry out carbon and energy metabolism in the host, and how they obtain or use key nutrients for replication and immune evasion. PMID:27337445

  20. Transport Policy and Social Inclusion

    Directory of Open Access Journals (Sweden)

    Miriam Ricci

    2016-06-01

    Full Text Available ‘Transport-related Social inclusion’ is a specific naming of the complex set of interrelationships within which accessibility plays an important role in whether a citizen achieves the level of participation in socioeconomic life that he or she seeks. It has its origins in the United Kingdom of the early 2000s, but the diversity of theoretical perspectives, research methods and practical focus shown by the contributions to the present issue on this theme bears witness to the evolution and translation this concept and term has undergone over more than a decade. Nine papers are presented, concerning applications of the concept in three continents, and including some of the poorest and richest per capita income countries on the globe. As well as developing and applying the multi-faceted theories of the processes of exclusion and techniques for the quantitative identification of inclusion, they consider important topics such as the treatment of the less abled and more frail members of society when on the move and the potential for new technological design methods and practical solutions either to enhance inclusion or deepen inequality in our societies. Collectively their conclusions reinforce the message that social exclusion remains multi-dimensional, relational and dynamic, located both in the circumstances of the excluded individual as well as in the processes, institutions and structures that permeate wider society.

  1. Semi-inclusive DIS: Factorization

    Energy Technology Data Exchange (ETDEWEB)

    Yuan, Feng

    2008-12-10

    In this talk, we will present a QCD factorization theorem for the semi-inclusive deep-inelastic scattering with hadrons in the current fragmentation region detected at low transverse momentum. There has been considerable experimental and theoretical interest in semi-inclusive hadron production in deep inelastic scattering (SIDIS) processes. For example, by studying the polarized and unpolarized SIDIS, one will be able to identify the sea quark distribution and polarization in nucleon, and the experimental results from the HERMES collaboration have revealed nontrivial sea structure in nucleon [1]. More recently, SIDIS opened a new window to study the transverse momentum dependent (TMD) parton distributions and fragmentation functions from the low transverse momentum hadron production. The transverse momentum distribution of the final state hadron is directly related to the transverse momentum dependence of the parton distributions and fragmentation. These studies will provide new opportunities to explore the partonic structure of nucleon, especially the three-dimension distribution of partons inside nucleon. The DIS experiments, including HERMES, COMPAS, and JLab Hall B collaborations, have studied various azimuthal asymmetries in SIDIS. In particular, the HERMES collaboration found sizable single spin asymmetries in these processes involving nontrivial QCD effects and hadron structure.

  2. The Bacterial Growth Curve.

    Science.gov (United States)

    Paulton, Richard J. L.

    1991-01-01

    A procedure that allows students to view an entire bacterial growth curve during a two- to three-hour student laboratory period is described. Observations of the lag phase, logarithmic phase, maximum stationary phase, and phase of decline are possible. A nonpathogenic, marine bacterium is used in the investigation. (KR)

  3. Bacterial fingerprints across Europe

    NARCIS (Netherlands)

    Glasner, Corinna

    2014-01-01

    Bacterial pathogens, such as Staphylococcus aureus and carbapenemase-producing Enterobacteriaceae (CPE), impose major threats to human health worldwide. Both have a ‘Jekyll & Hyde’ character, since they can be present as human commensals, but can also become harmful invasive pathogens especially

  4. EDITORIAL SPONTANEOUS BACTERIAL PERITONITIS ...

    African Journals Online (AJOL)

    hi-tech

    Spontaneous bacterial peritonitis (SBP) frequent]y occurs in patients with liver cirrhosis and ascites. It is defined as an infection of previously sterile ascitic fluid without any demonstrable intrabdominal source of infection. It is now internationally agreed that a polymorphonuclear (PMN) cell count in the ascitic fluid of over 250 ...

  5. Bacterial membrane proteomics.

    Science.gov (United States)

    Poetsch, Ansgar; Wolters, Dirk

    2008-10-01

    About one quarter to one third of all bacterial genes encode proteins of the inner or outer bacterial membrane. These proteins perform essential physiological functions, such as the import or export of metabolites, the homeostasis of metal ions, the extrusion of toxic substances or antibiotics, and the generation or conversion of energy. The last years have witnessed completion of a plethora of whole-genome sequences of bacteria important for biotechnology or medicine, which is the foundation for proteome and other functional genome analyses. In this review, we discuss the challenges in membrane proteome analysis, starting from sample preparation and leading to MS-data analysis and quantification. The current state of available proteomics technologies as well as their advantages and disadvantages will be described with a focus on shotgun proteomics. Then, we will briefly introduce the most abundant proteins and protein families present in bacterial membranes before bacterial membrane proteomics studies of the last years will be presented. It will be shown how these works enlarged our knowledge about the physiological adaptations that take place in bacteria during fine chemical production, bioremediation, protein overexpression, and during infections. Furthermore, several examples from literature demonstrate the suitability of membrane proteomics for the identification of antigens and different pathogenic strains, as well as the elucidation of membrane protein structure and function.

  6. Seizures Complicating Bacterial Meningitis

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2004-09-01

    Full Text Available The clinical data of 116 patients, 1 month to <5 years of age, admitted for bacterial meningitis, and grouped according to those with and without seizures during hospitalization, were compared in a study at Buddhist Dalin Tzu Chi General Hospital, Chang Gung Memorial Hospital and other centers in Taiwan.

  7. Diagnosis of bacterial infection

    African Journals Online (AJOL)

    rapid and easy-to-use test for bacterial infections. Clearly, this is a very ... detect antigens or specific antibodies, e.g. group A streptococcal antigen testing can be employed to reduce antibiotic use. Culture-based tests are often ... White blood cell count 12 000 cells/mm³; or the presence of >10% ...

  8. Bacterial Meningitis Outcome

    OpenAIRE

    J Gordon Millichap

    1995-01-01

    The neurologic, psychological, and educational outcomes of bacterial meningitis in 130 children evaluated at a mean age of 8 years, and 6 years after their meningitis, are reported from the Department of Paediatrics and Clinical Epidemiology and Biostatistics Unit, University of Melbourne, and the Royal Children’s Hospital, Victoria, Australia.

  9. Measuring Quality in Inclusive Preschool Classrooms: Development and Validation of the Inclusive Classroom Profile (ICP)

    Science.gov (United States)

    Soukakou, Elena P.

    2012-01-01

    The purpose of this study was to develop and validate an observation measure designed to assess classroom quality in inclusive preschool programs, the Inclusive Classroom Profile (ICP). Developing the rating scale entailed systematic fieldwork in inclusive settings and review of the literature on preschool inclusion. Results from the validation…

  10. Corticosteroids for Bacterial Keratitis

    Science.gov (United States)

    Srinivasan, Muthiah; Mascarenhas, Jeena; Rajaraman, Revathi; Ravindran, Meenakshi; Lalitha, Prajna; Glidden, David V.; Ray, Kathryn J.; Hong, Kevin C.; Oldenburg, Catherine E.; Lee, Salena M.; Zegans, Michael E.; McLeod, Stephen D.; Lietman, Thomas M.; Acharya, Nisha R.

    2013-01-01

    Objective To determine whether there is a benefit in clinical outcomes with the use of topical corticosteroids as adjunctive therapy in the treatment of bacterial corneal ulcers. Methods Randomized, placebo-controlled, double-masked, multicenter clinical trial comparing prednisolone sodium phosphate, 1.0%, to placebo as adjunctive therapy for the treatment of bacterial corneal ulcers. Eligible patients had a culture-positive bacterial corneal ulcer and received topical moxifloxacin for at least 48 hours before randomization. Main Outcome Measures The primary outcome was best spectacle-corrected visual acuity (BSCVA) at 3 months from enrollment. Secondary outcomes included infiltrate/scar size, reepithelialization, and corneal perforation. Results Between September 1, 2006, and February 22, 2010, 1769 patients were screened for the trial and 500 patients were enrolled. No significant difference was observed in the 3-month BSCVA (−0.009 logarithm of the minimum angle of resolution [logMAR]; 95% CI, −0.085 to 0.068; P = .82), infiltrate/scar size (P = .40), time to reepithelialization (P = .44), or corneal perforation (P > .99). A significant effect of corticosteroids was observed in subgroups of baseline BSCVA (P = .03) and ulcer location (P = .04). At 3 months, patients with vision of counting fingers or worse at baseline had 0.17 logMAR better visual acuity with corticosteroids (95% CI, −0.31 to −0.02; P = .03) compared with placebo, and patients with ulcers that were completely central at baseline had 0.20 logMAR better visual acuity with corticosteroids (−0.37 to −0.04; P = .02). Conclusions We found no overall difference in 3-month BSCVA and no safety concerns with adjunctive corticosteroid therapy for bacterial corneal ulcers. Application to Clinical Practice Adjunctive topical corticosteroid use does not improve 3-month vision in patients with bacterial corneal ulcers. PMID:21987582

  11. An Arendtian perspective on inclusive education

    DEFF Research Database (Denmark)

    Korsgaard, Morten Timmermann

    2016-01-01

    Inclusive education currently appears to be undergoing a crisis and re- examination. This paper presents a new approach to thinking about inclusiveness in the school context. Many positions within inclusive education seem to take political, social and ethical perspectives as a starting point, which...... has allowed inclusive movements and initiatives around the world to succumb to neo-liberal policy-making and has neglected the development of an educational vocabulary that is theoretically and conceptually appropriate for confronting teachers’ central concerns regarding inclusive practices....... The concepts of suspension, bearing with strangers and enlarged thought inspired by Hannah Arendt provide a basis for a re-imagining of inclusive education and for outlining a future school in which inclusiveness is embedded in the very way we think and position ourselves as teachers and pupils...

  12. A review of social inclusion measures.

    Science.gov (United States)

    Coombs, Tim; Nicholas, Angela; Pirkis, Jane

    2013-10-01

    Social inclusion is crucial to mental health and well-being and is emphasised in Australia's Fourth National Mental Health Plan. There is a recognition that a measure of social inclusion would complement the suite of outcome measures that is currently used in public sector mental health services. This paper is an initial scope of candidate measures of social inclusion and considers their suitability for this purpose. We identified potential measures through searches of PsycINFO and Medline and a more general Internet search. We extracted descriptive and evaluative information on each measure identified and compared this information with a set of eight criteria. The criteria related to the measure's inclusion of four domains of social inclusion outlined in Australia's Fourth National Mental Health Plan, its usability within the public mental health sector and its psychometric properties. We identified 10 candidate measures of social inclusion: the Activity and Participation Questionnaire (APQ-6); the Australian Community Participation Questionnaire (ACPQ); the Composite Measure of Social Inclusion (CMSI); the EMILIA Project Questionnaire (EPQ); the Evaluating Social Inclusion Questionnaire (ESIQ); the Inclusion Web (IW); the Social and Community Opportunities Profile (SCOPE); the Social Inclusion Measure (SIM); the Social Inclusion Questionnaire (SIQ); and the Staff Survey of Social Inclusion (SSSI). After comparison with the eight review criteria, we determined that the APQ-6 and the SCOPE-short form show the most potential for further testing. Social inclusion is too important not to measure. This discussion of individual-level measures of social inclusion provides a springboard for selecting an appropriate measure for use in public sector mental health services. It suggests that there are two primary candidates, but neither of these is quite fit-for-purpose in their current form. Further exploration will reveal whether one of these is suitable, whether another

  13. Phages in the Human Body.

    Science.gov (United States)

    Navarro, Ferran; Muniesa, Maite

    2017-01-01

    Bacteriophages, viruses that infect bacteria, have re-emerged as powerful regulators of bacterial populations in natural ecosystems. Phages invade the human body, just as they do other natural environments, to such an extent that they are the most numerous group in the human virome. This was only revealed in recent metagenomic studies, despite the fact that the presence of phages in the human body was reported decades ago. The influence of the presence of phages in humans has yet to be evaluated; but as in marine environments, a clear role in the regulation of bacterial populations could be envisaged, that might have an impact on human health. Moreover, phages are excellent vehicles of genetic transfer, and they contribute to the evolution of bacterial cells in the human body by spreading and acquiring DNA horizontally. The abundance of phages in the human body does not pass unnoticed and the immune system reacts to them, although it is not clear to what extent. Finally, the presence of phages in human samples, which most of the time is not considered, can influence and bias microbiological and molecular results; and, in view of the evidences, some studies suggest that more attention needs to be paid to their interference.

  14. Index of Financial Inclusion – A measure of financial sector inclusiveness

    OpenAIRE

    Mandira Sarma

    2012-01-01

    The promotion of an inclusive financial system is a policy priority in many countries. While the importance of financial inclusion is widely recognized, the literature lacks a comprehensive measure that can be used to measure the extent of financial inclusion across economies. This paper attempts to fill this gap by proposing an index of financial inclusion (IFI). The proposed IFI captures information on various dimensions of financial inclusion in a single number lying between 0 and 1, where...

  15. Irradiation history of meteoritic inclusions

    DEFF Research Database (Denmark)

    Wielandt, Daniel Kim Peel

    be coproduced at approximately the value found in CAIs. The required fluence levels can be reached in as little as half a year of particle irradiation at protosolar analog flux levels. We thus conclude that local charged particle irradiation played a significant if not dominating role in forming the 10Be and 41...... and local particle irradiation. If local particle irradiation was powerful, it may have left telltale signs in meteoritic inclusions that can constrain the conditions under which they were formed or stored prior to the formation of chondrites. The underlying research effort relates to answering the larger...... in the early solar system. We demonstrate novel techniques for measuring the isotopic composition of K, and show how such measurements can be related to the irradiation histories of meteoritic materials. We also show how potassium isotope measurement can complement measurements of 10Be, a proven spallogenic...

  16. Inclusion, children's groups, music therapy

    DEFF Research Database (Denmark)

    Holck, Ulla; Jacobsen, Stine Lindahl

    2016-01-01

    a delightful build-up of tension or soothes us, and its narrative character gives rise to mental imagery or memories. Music brings people together and helps build communities across languages and common divides. And – not least – music captures children’s immediate attention, so when the music starts, so do...... selected six case vignettes from Denmark to illustrate how musical interplay can promote attention, concentration, emotional attachment and social skills in groups and, in the last of our case vignettes, also the quality of parent-child interactions. The specific cases were selected because they involve...... portrayal of the qualities of musical interplay that promotes well-being in group settings and, thus, the inclusion of vulnerable students. Therefore, we open the chapter with a focus on musicality and on the importance of applying a musical approach in relation to the children....

  17. How Bureaucracy Promotes Inclusive Organizing

    DEFF Research Database (Denmark)

    Holck, Lotte

    and informality struggle with membership perceptions of partiality and unfairness on the basis of ethnicity and professional background, then the bureaucratic structured organization characterized by structural stability and formality is more successful in generating the necessary membership perception fairness......Diversity literature in general and Feminist in particular have long promoted alternatives to bureaucracy on the premise that this form of governance is far from gender- and race-neutral, and that inclusive organizing necessitate a flatter, decentralized and more ‘organic’ set-up (Ferguson 1984......, Martin P.Y 1987, Ashcraft 2001, 2006, 2013, Acker 1990, Ghorashi & Sabelis 2012, Dai 2014). My ethnographic research in two case organizations both championed for their diversity work in a Danish context, shows a different pattern: While the organization characterized by structural flexibility...

  18. Interprofessional development in inclusive schools

    DEFF Research Database (Denmark)

    Hansen, Britt Blaabjerg; Højholdt, Andy; Arndal, Lars Stubbe

    practices/the need to establish collaborative practices between the different groups of professionals working at a school (Downes 2011, Anderson-Butcher 2004). To ensure all students succeed at school, teachers and child and youth educators need to collaborate more effectively, particularly when it comes...... educators, both initiatives of which support the subject-oriented education at school. These changes have challenged the way [education/school] professionals work and how they see their own and each other's roles in the school system (EVA 2013). The changes have also increased the need for collaborative...... required to work with these students. This paper describes the challenges [education/school] professionals (teachers and child and youth educators) face in striving to develop inclusive school practice, and how these challenges relate to differences in skills and competencies between the professions...

  19. Inclusive radiative J/ψ decays

    International Nuclear Information System (INIS)

    Besson, D.; Pedlar, T. K.; Cronin-Hennessy, D.; Gao, K. Y.; Hietala, J.; Kubota, Y.; Klein, T.; Lang, B. W.; Poling, R.; Scott, A. W.; Zweber, P.; Dobbs, S.; Metreveli, Z.; Seth, K. K.; Tomaradze, A.; Libby, J.; Powell, A.; Wilkinson, G.; Ecklund, K. M.; Love, W.

    2008-01-01

    Using data taken with the CLEO-c detector at the Cornell Electron Storage Ring, we have investigated the direct-photon momentum spectrum in the decay J/ψ(1S)→γgg, via the ''tagged'' process: e + e - →ψ(2S); ψ(2S)→J/ψπ + π - ; J/ψ→γ+X. Including contributions from two-body radiative decay processes, we find the ratio of the inclusive direct-photon branching fraction to that of the dominant three-gluon branching fraction [R γ =B(ggγ)/B(ggg)] to be R γ =0.137±0.001±0.016±0.004, where the errors shown are statistical, systematic, and the model-dependent uncertainty related to the extrapolation to zero photon energy. The shape of the scaled photon energy spectrum in J/ψ→ggγ is observed to be very similar to that of Υ→ggγ. The R γ value obtained is roughly consistent with that expected by a simple quark-charge scaling [R γ ∼(q c /q b ) 2 ] of the value determined at the Υ(1S), but somewhat higher than the value expected from the running of the strong coupling constant.

  20. Inclusive breakup of three-fragment weakly bound nuclei

    Energy Technology Data Exchange (ETDEWEB)

    Carlson, B.V.; Frederico, T. [Instituto Tecnológico de Aeronáutica, DCTA, 12.228-900 São José dos Campos, SP (Brazil); Hussein, M.S., E-mail: hussein@if.usp.br [Instituto Tecnológico de Aeronáutica, DCTA, 12.228-900 São José dos Campos, SP (Brazil); Instituto de Estudos Avançados, Universidade de São Paulo, C.P. 72012, 05508-970 São Paulo, SP (Brazil); Instituto de Física, Universidade de São Paulo, C.P. 66318, 05314-970 São Paulo, SP (Brazil)

    2017-04-10

    The inclusive breakup of three-fragment projectiles is discussed within a four-body spectator model. Both the elastic breakup and the non-elastic breakup are obtained in a unified framework. Originally developed in the 80's for two-fragment projectiles such as the deuteron, in this paper the theory is successfully generalized to three-fragment projectiles. The expression obtained for the inclusive cross section allows the extraction of the incomplete fusion cross section, and accordingly generalizes the surrogate method to cases such as (t, p) and (t, n) reactions. It is found that two-fragment correlations inside the projectile affect in a conspicuous way the elastic breakup cross section. The inclusive non-elastic breakup cross section is calculated and is found to contain the contribution of a three-body absorption term that is also strongly influenced by the two-fragment correlations. This latter cross section contains the so-called incomplete fusion where more than one compound nuclei are formed. Our theory describes both stable weakly bound three-fragment projectiles and unstable ones such as the Borromean nuclei.

  1. Extraterrestrial diamond inclusions reveal a Mars-sized planet

    Science.gov (United States)

    Nabiei, F.; Badro, J.; Dennenwaldt, T.; Oveisi, E.; Cantoni, M.; Hébert, C.; El Goresy, A.; Barrat, J. A.; Gillet, P.

    2017-12-01

    Ureilites are achondritic meteorites, distinct by their high carbon content. Carbon dominantly exists as graphite and diamond forming veins between olivine and pyroxene grains in the matrix. The presence of diamonds is, often, attributed to the shock-induced transformation from graphite. However, a recent study (Miyahara et al. 2016) on Almahata Sitta MS-170 mainly based on the large size of diamonds suggested their growth in the static high pressure conditions of planetary interior. We prepared five thin sections by the focused ion beam (FIB) from the diamonds in Almahatta Sitta MS-170 for transmission electron microscopy (TEM) analysis. Abundance of dislocations, stacking faults and {111} twinnings indicate significant deformation. Moreover, large numbers of inclusions are found in diamonds. Electron energy-loss spectroscopy (EELS) showed that when a twinning is intersected with an inclusion, it transforms to graphite. This together with other morphological and crystallographical characteristics of the graphite and diamond phases point to the shock-induced transformation of diamond to graphite. Energy dispersive X-ray (EDX) spectroscopy and electron diffraction were used to chemically and structurally characterize the diamond inclusions. Most of the inclusions are Fe-Ni-S-P type up to 60 nm in diameter, each consisting of three phases: kamacite (Fe, Ni), troilite (FeS) and Schreibersite ((Fe,Ni)3P). The inclusions always have euhedral shape indicating the existence of a parent phase that later broke down. Chemical analysis of complete inclusions (identified with electron tomography) agrees with the stoichiometric (Fe0.93,Ni0.07)3(S0.88,P0.12) phase that only forms above 21 GPa (Gu et al. 2016). The ureilite parent body (UPB) needs to be at least about Mars-sized to generate such a pressure at its core-mantle boundary. This is in the same size range estimated for the planetary embryos forming early in the Solar System. Moreover, although the UPB did not go through

  2. Radiometric detection of bacterial metabolism

    International Nuclear Information System (INIS)

    Camargo, E.E.; Wagner Junior, H.N.

    1979-01-01

    The measurement of 14 CO 2 produced by the bacterial oxidation of labelled compounds is discussed as a means of evaluating the bacterial metabolism. The following items are discussed:automated radiometric detection, types of graphs, clinical applications of the radiometric system and influential factors. Complementary studies on bacterial assimilation of substances are presented. (M.A.) [pt

  3. Removal of Inclusions from Aluminum Through Filtration

    Science.gov (United States)

    Damoah, Lucas Nana Wiredu; Zhang, Lifeng

    2010-08-01

    Filtration experiments were carried out using both an AlF3 slurry-coated and an uncoated Al2O3 ceramic foam filter to study the removal of nonmetallic inclusions and impurity elements. The results showed that the 30-ppi ceramic foam filter removed up to 85 pct inclusions from aluminum. Several pictures of two- and three-dimensional morphologies of both nonmetallic and intermetallics inclusions also have been presented. The following contributing mechanisms for the removal of nonmetallic inclusions in the deep-bed filtration mode are proposed: (1) collision with walls and interception effect and (2) the formation of both intermetallic and nonmetallic inclusion bridges during filtration. Fluid dynamics modeling of inclusion attachment to the filter walls showed that most inclusions, especially those with larger sizes, are entrapped at the upper part of the filter, whereas smaller inclusions are dispersed well throughout the filter. The calculated inclusions removal fractions for the 30-ppi filter showed that almost all inclusions >125 μm are removed, and inclusions ~5 μm in size are removed up to 85 pct. The interfacial energy between two collided same-size inclusions was calculated, indicating that a strong clustering of inclusions may result within the filter window. Magnesium impurities were removed up to 86 pct by the AlF3 slurry-coated filter. The filter acted in active filtration mode in addition to the contribution of the air oxidation of dissolved [Mg], which was calculated to be 13 pct. The total mass transfer coefficient of dissolved [Mg] to the reaction interface was calculated to be 1.15 × 10-6 m/s.

  4. Perceived and Actual Social Discrimination: The Case of Overweight and Social Inclusion

    OpenAIRE

    Hartung, Freda-Marie; Renner, Britta

    2013-01-01

    The present study examined the correspondence between perceived and actual social discrimination of overweight people. In total, 77 first-year students provided self-ratings about their height, weight, and perceived social inclusion. To capture actual social inclusion, each participant nominated those fellow students (a) she/he likes and dislikes and (b) about whom she/he is likely to hear social news. Students with lower Body Mass Index (BMI) felt socially included, irrespective of their act...

  5. Bacterial Cell Wall Components

    Science.gov (United States)

    Ginsberg, Cynthia; Brown, Stephanie; Walker, Suzanne

    Bacterial cell-surface polysaccharides cells are surrounded by a variety of cell-surface structures that allow them to thrive in extreme environments. Components of the cell envelope and extracellular matrix are responsible for providing the cells with structural support, mediating intercellular communication, allowing the cells to move or to adhere to surfaces, protecting the cells from attack by antibiotics or the immune system, and facilitating the uptake of nutrients. Some of the most important cell wall components are polysaccharide structures. This review discusses the occurrence, structure, function, and biosynthesis of the most prevalent bacterial cell surface polysaccharides: peptidoglycan, lipopolysaccharide, arabinogalactan, and lipoarabinomannan, and capsular and extracellular polysaccharides. The roles of these polysaccharides in medicine, both as drug targets and as therapeutic agents, are also described.

  6. Bacterial meningitis in Nottingham.

    OpenAIRE

    Ispahani, P.

    1983-01-01

    Records of 171 cases of bacterial meningitis admitted to Nottingham hospitals from January 1974 to June 1980 were reviewed. The distribution of organisms producing meningitis and the factors influencing mortality in different age groups were assessed. Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae accounted for 69% of all proven cases. The overall mortality was 26% being lowest in patients with meningococcal meningitis (0%) and highest in those with pneumococcal m...

  7. Neglected bacterial zoonoses.

    Science.gov (United States)

    Chikeka, I; Dumler, J S

    2015-05-01

    Bacterial zoonoses comprise a group of diseases in humans or animals acquired by direct contact with or by oral consumption of contaminated animal materials, or via arthropod vectors. Among neglected infections, bacterial zoonoses are among the most neglected given emerging data on incidence and prevalence as causes of acute febrile illness, even in areas where recognized neglected tropical diseases occur frequently. Although many other bacterial infections could also be considered in this neglected category, five distinct infections stand out because they are globally distributed, are acute febrile diseases, have high rates of morbidity and case fatality, and are reported as commonly as malaria, typhoid or dengue virus infections in carefully designed studies in which broad-spectrum diagnoses are actively sought. This review will focus attention on leptospirosis, relapsing fever borreliosis and rickettsioses, including scrub typhus, murine typhus and spotted fever group rickettsiosis. Of greatest interest is the lack of distinguishing clinical features among these infections when in humans, which confounds diagnosis where laboratory confirmation is lacking, and in regions where clinical diagnosis is often attributed to one of several perceived more common threats. As diseases such as malaria come under improved control, the real impact of these common and under-recognized infections will become evident, as will the requirement for the strategies and allocation of resources for their control. Copyright © 2015 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  8. Bacterial growth kinetics

    International Nuclear Information System (INIS)

    Boonkitticharoen, V.; Ehrhardt, J.C.; Kirchner, P.T.

    1989-01-01

    Quantitative measurement of bacterial growth may be made using a radioassay technique. This method measures, by scintillation counting, the 14 CO 2 derived from the bacterial metabolism of a 14 C-labeled substrate. Mathematical growth models may serve as reliable tools for estimation of the generation rate constant (or slope of the growth curve) and provide a basis for evaluating assay performance. Two models, i.e., exponential and logistic, are proposed. Both models yielded an accurate fit to the data from radioactive measurement of bacterial growth. The exponential model yielded high precision values of the generation rate constant, with an average relative standard deviation of 1.2%. Under most conditions the assay demonstrated no changes in the slopes of growth curves when the number of bacteria per inoculation was changed. However, the radiometric assay by scintillation method had a growth-inhibiting effect on a few strains of bacteria. The source of this problem was thought to be hypersensitivity to trace amounts of toluene remaining on the detector

  9. Indonesian teachers' epistemological beliefs and inclusive education.

    Science.gov (United States)

    Sheehy, Kieron; Budiyanto; Kaye, Helen; Rofiah, Khofidotur

    2017-01-01

    A growing number of children with intellectual disabilities attend inclusive schools in Indonesia. Previous research has suggested that teachers' type of school and experience influences their beliefs about inclusive education. This research collected questionnaire data from 267 Indonesian teachers and compared the responses from those working in inclusive, special and regular schools regarding their epistemological and pedagogical beliefs. The results showed that teachers in inclusive schools expressed stronger social constructivist beliefs than those in other schools. However, it was teachers' epistemological beliefs, rather than their type of school or experience, which were the significant predictor of their beliefs about inclusive education. The findings suggest that international epistemological research needs to have a more nuanced view of constructivist models of learning to better understand and inform how inclusive pedagogy is being enacted in different contexts.

  10. Inclusive Islamic financial planning : a conceptual framework.

    OpenAIRE

    Ahmed, H.; Salleh, H.

    2016-01-01

    Purpose To develop a conceptual framework of inclusive Islamic Financial Planning (IFP) by combining the traditional Islamic institutions of zakat and awqaf with contemporary notions of financial planning, financial inclusion and financial literacy that caters to the short-term and long-term financial goals of the poor. Design/methodology/approach Being a conceptual article, an inclusive Islamic Financial Planning framework is described, analyzed and developed by integrating modern ...

  11. Measuring Financial Inclusion: A Multidimensional Index

    OpenAIRE

    Noelia Camara; David Tuesta

    2014-01-01

    We rely on demand and supply-side information to measure the extent of financial inclusion at country level for eighty-two developed and less-developed countries. We postulate that the degree of financial inclusion is determined by three dimensions: usage, barriers and access to financial inclusion. Weights assigned to the dimensions are determined endogenously by employing a two-stage Principal Component Analysis. Our composite index offers a comprehensive measure of the degree of financial ...

  12. DEMONETIZATION A STEP TOWARDS MODERN FINANCIAL INCLUSION

    OpenAIRE

    Dharini Raje Sisodia; Akanksha Kapoor

    2017-01-01

    India creating a high expectation for the world to become economic leader. From quite a while Indian took Financial Inclusion as serious course of Business as a development tool and encouraging result are been observed. In order to have strong financial inclusion serious action are required. In this era of technology with largest youth population in India, this is the best time to strike the chords of modern financial inclusion. This paper will take up the issues related to demonetization and...

  13. Inclusive Fitness Maximization:An Axiomatic Approach

    OpenAIRE

    Okasha, Samir; Weymark, John; Bossert, Walter

    2014-01-01

    Kin selection theorists argue that evolution in social contexts will lead organisms to behave as if maximizing their inclusive, as opposed to personal, fitness. The inclusive fitness concept allows biologists to treat organisms as akin to rational agents seeking to maximize a utility function. Here we develop this idea and place it on a firm footing by employing a standard decision-theoretic methodology. We show how the principle of inclusive fitness maximization and a related principle of qu...

  14. Inclusive central region in perturbative Reggeon calculus

    International Nuclear Information System (INIS)

    Pajares, C.; Pascual, R.

    1976-01-01

    The single-particle inclusive cross section and the correlation function are studied in the perturbative approach to Gribov's Reggeon calculus; the leading contributions to both functions are evaluated. The large energy rise of the inclusive cross section appears as a consequence of the Pomerons having an intercept larger than 1. The same set of parameters which describes correctly the cross-section data and the triple-Regge region also describes the inclusive data in the central region

  15. The problems of Russian inclusive education

    Directory of Open Access Journals (Sweden)

    Kurmyshova O.A.

    2017-02-01

    Full Text Available the article is devoted to problems of Russian inclusive education. Based on the results of theoretical analysis the author raises some questions and their solutions may help to increase the effectiveness of the activity of inclusive educational organizations, provide harmonization of relations of inclusive education members and that will guarantee a successful integration of children with disabilities and special needs in the society.

  16. Body punk

    DEFF Research Database (Denmark)

    Mogensen, Kevin

    BODYPUNK - A Treatise on male body builders and the meaning of the body in the shadow of an Anti Doping Campaign Based on a qualitative study, the thesis investigates the visual representation of the male bodybuilder found in the national anti doping campaign: ‗ "The hunt has begun" along...... with an analysis of the embodied meaning of men‘s bodybuilding....

  17. Making the case for inclusive design.

    Science.gov (United States)

    Waller, Sam; Bradley, Mike; Hosking, Ian; Clarkson, P John

    2015-01-01

    This paper describes the University of Cambridge, Engineering Design Centre's (EDC) case for inclusive design, based on 10 years of research, promotion and knowledge transfer. In summary, inclusive design applies an understanding of customer diversity to inform decisions throughout the development process, in order to better satisfy the needs of more people. Products that are more inclusive can reach a wider market, improve customer satisfaction and drive business success. The rapidly ageing population increases the importance of this approach. The case presented here has helped to convince BT, Nestlé and others to adopt an inclusive approach. Copyright © 2013 Elsevier Ltd and The Ergonomics Society. All rights reserved.

  18. Diversity at work the practice of inclusion

    CERN Document Server

    Deane, Barbara R

    2014-01-01

    This book outlines the key issues involved in framing, designing, and implementing inclusion initiatives for organizations and groups. It offers ideas for helping individuals develop competencies for inclusion. It shows how to apply the practices of inclusion and provides a unified model by employing diverse voices to address a range of related topics in multiple contexts. It also contains examples of how diversity and inclusion has worked in a variety of settings. The book is includes information from topic experts, including internal and external change agents and academics.

  19. Inclusive education foreign experience and Russian realities

    Directory of Open Access Journals (Sweden)

    Natalya V. Borodina

    2011-04-01

    Full Text Available The article analyses social processes, effecting inclusive education development in countries with liberal welfare model and problems of integrated education development in Russia.

  20. Solid-state nuclear magnetic resonance (NMR) spectroscopy of human immunodeficiency virus gp41 protein that includes the fusion peptide: NMR detection of recombinant Fgp41 in inclusion bodies in whole bacterial cells and structural characterization of purified and membrane-associated Fgp41.

    Science.gov (United States)

    Vogel, Erica P; Curtis-Fisk, Jaime; Young, Kaitlin M; Weliky, David P

    2011-11-22

    Human immunodeficiency virus (HIV) infection of a host cell begins with fusion of the HIV and host cell membranes and is mediated by the gp41 protein, a single-pass integral membrane protein of HIV. The 175 N-terminal residues make up the ectodomain that lies outside the virus. This work describes the production and characterization of an ectodomain construct containing the 154 N-terminal gp41 residues, including the fusion peptide (FP) that binds to target cell membranes. The Fgp41 sequence was derived from one of the African clade A strains of HIV-1 that have been less studied than European/North American clade B strains. Fgp41 expression at a level of ~100 mg/L of culture was evidenced by an approach that included amino acid type (13)CO and (15)N labeling of recombinant protein and solid-state NMR (SSNMR) spectroscopy of lyophilized whole cells. The approach did not require any protein solubilization or purification and may be a general approach for detection of recombinant protein. The purified Fgp41 yield was ~5 mg/L of culture. SSNMR spectra of membrane-associated Fgp41 showed high helicity for the residues C-terminal of the FP. This was consistent with a "six-helix bundle" (SHB) structure that is the final gp41 state during membrane fusion. This observation and negligible Fgp41-induced vesicle fusion supported a function for SHB gp41 of membrane stabilization and fusion arrest. SSNMR spectra of residues in the membrane-associated FP provided evidence of a mixture of molecular populations with either helical or β-sheet FP conformation. These and earlier SSNMR data strongly support the existence of these populations in the SHB state of membrane-associated gp41. © 2011 American Chemical Society

  1. Bacterial species isolated from water bodies in Rivers and Bayelsa ...

    African Journals Online (AJOL)

    Methods: A total of twenty water samples were collected from rivers and cultured for the various microorganisms.The total heterotrophic counts were determined by Standard Plate Count (SPC) method. Salmonella and Vibrio species were enumerated by an enrichment procedure in Selenite F broth and Alkaline Peptone ...

  2. Broad recruitment of mGBP family members to Chlamydia trachomatis inclusions.

    Directory of Open Access Journals (Sweden)

    Valesca Lindenberg

    Full Text Available Chlamydia, the most common sexually transmitted pathogen, is an exquisitely adapted Gram-negative obligate intracellular bacterium. Intracellular Chlamydia trachomatis replicate in a specialized vacuole, termed inclusion, which shields the bacterium from antimicrobial immunity of the host cells and acts as a signalling interface. Previously it was shown that members of the interferon induced guanylate binding protein (mGBP family, in particular murine GBP1 and mGBP2, were found to accumulate at the bacterial inclusions, similar to previously published recruitment of GBPs to the parasitophorous vacuole of Toxoplasma gondii. Here, we provide a wide comparison of mGBPs roles within the host cell in the context of Chlamydia and Toxoplasma infection. By confocal microscopy on fixed and living infected cells we show localization of mGBP3, mGBP6, mGBP7, mGBP9, and mGBP10, in addition to mGBP1 and mGBP2, at chlamydia inclusions. In time lapse videos using GFP expressing Chlamydia we show rapid and transient dynamics of mGBP9 accumulation onto chlamydia inclusions. Taken together this study reveals a broad activation of mGBP recruitment towards Chlamydia trachomatis inclusions after infection and provides evidence for time limited action of mGBP9 at the chlamydia inclusion.

  3. Bacterial Succession on Rat Carcasses and Applications for PMI Estimation.

    Science.gov (United States)

    Zhang, Lin; Guo, Juan-juan; Telet-Siyit; Peng, Yu-long; Xie, Dan; Guo, Ya-dong; Yan, Jie; Zha, Lagabaiyila; Cai, Ji-feng

    2016-02-01

    Abstract: To investigate the bacterial succession on rat carcasses and to evaluate the use of bacterial succession for postmortem interval (PMI) estimation. Adult female SD rat remains were placed in carton boxes. The bacterial colonization of circumocular skin, mouth and vagina was collected to be identified using culture-dependent biochemical methods. The changes in community composition were regularly documented. The bacterial succession in three habitats showed that Staphylococcus and Neisseria were predominated in early PMI, especially Staphylococcus aureus and Neisseria lactamica in 6 hours after death. Lactobacillus casei developed on the 3-4 days regularly, and kept stable at a certain level in late PMI. The involvement of normal and putrefactive bacteria in three body habitats of rat remains can be used for PMI estimation.

  4. Radiology of bacterial pneumonia

    Energy Technology Data Exchange (ETDEWEB)

    Vilar, Jose E-mail: vilar_jlu@gva.es; Domingo, Maria Luisa; Soto, Cristina; Cogollos, Jonathan

    2004-08-01

    Bacterial pneumonia is commonly encountered in clinical practice. Radiology plays a prominent role in the evaluation of pneumonia. Chest radiography is the most commonly used imaging tool in pneumonias due to its availability and excellent cost benefit ratio. CT should be used in unresolved cases or when complications of pneumonia are suspected. The main applications of radiology in pneumonia are oriented to detection, characterisation and follow-up, especially regarding complications. The classical classification of pneumonias into lobar and bronchial pneumonia has been abandoned for a more clinical classification. Thus, bacterial pneumonias are typified into three main groups: Community acquired pneumonia (CAD), Aspiration pneumonia and Nosocomial pneumonia (NP).The usual pattern of CAD is that of the previously called lobar pneumonia; an air-space consolidation limited to one lobe or segment. Nevertheless, the radiographic patterns of CAD may be variable and are often related to the causative agent. Aspiration pneumonia generally involves the lower lobes with bilateral multicentric opacities. Nosocomial Pneumonia (NP) occurs in hospitalised patients. The importance of NP is related to its high mortality and, thus, the need to obtain a prompt diagnosis. The role of imaging in NP is limited but decisive. The most valuable information is when the chest radiographs are negative and rule out pneumonia. The radiographic patterns of NP are very variable, most commonly showing diffuse multifocal involvement and pleural effusion. Imaging plays also an important role in the detection and evaluation of complications of bacterial pneumonias. In many of these cases, especially in hospitalised patients, chest CT must be obtained in order to better depict these associate findings.

  5. Radiology of bacterial pneumonia

    International Nuclear Information System (INIS)

    Vilar, Jose; Domingo, Maria Luisa; Soto, Cristina; Cogollos, Jonathan

    2004-01-01

    Bacterial pneumonia is commonly encountered in clinical practice. Radiology plays a prominent role in the evaluation of pneumonia. Chest radiography is the most commonly used imaging tool in pneumonias due to its availability and excellent cost benefit ratio. CT should be used in unresolved cases or when complications of pneumonia are suspected. The main applications of radiology in pneumonia are oriented to detection, characterisation and follow-up, especially regarding complications. The classical classification of pneumonias into lobar and bronchial pneumonia has been abandoned for a more clinical classification. Thus, bacterial pneumonias are typified into three main groups: Community acquired pneumonia (CAD), Aspiration pneumonia and Nosocomial pneumonia (NP).The usual pattern of CAD is that of the previously called lobar pneumonia; an air-space consolidation limited to one lobe or segment. Nevertheless, the radiographic patterns of CAD may be variable and are often related to the causative agent. Aspiration pneumonia generally involves the lower lobes with bilateral multicentric opacities. Nosocomial Pneumonia (NP) occurs in hospitalised patients. The importance of NP is related to its high mortality and, thus, the need to obtain a prompt diagnosis. The role of imaging in NP is limited but decisive. The most valuable information is when the chest radiographs are negative and rule out pneumonia. The radiographic patterns of NP are very variable, most commonly showing diffuse multifocal involvement and pleural effusion. Imaging plays also an important role in the detection and evaluation of complications of bacterial pneumonias. In many of these cases, especially in hospitalised patients, chest CT must be obtained in order to better depict these associate findings

  6. Signifying Bodies

    DEFF Research Database (Denmark)

    of biosemiosis connect signifying bodies with their natural surroundings, cultural activities and subjective experiences. Health stretches all the way from the ecosocial surroundings, through the skin and into the self-organizing processes of every living cell. Signifying Bodies lays out a new approach to health...... and health care. Eschewing all forms of dualism, the authors emphasise the interdependency of how we act, think, feel and function. They advocate a relational turn in health care, in which bodies live and learn from suffering and care. In this view, health is inseparable from both living beings...

  7. Bacterial Degradation of Pesticides

    DEFF Research Database (Denmark)

    Knudsen, Berith Elkær

    This PhD project was carried out as part of the Microbial Remediation of Contaminated Soil and Water Resources (MIRESOWA) project, funded by the Danish Council for Strategic Research (grant number 2104-08-0012). The environment is contaminated with various xenobiotic compounds e.g. pesticides......D student, to construct fungal-bacterial consortia in order to potentially stimulate pesticide degradation thereby increasing the chance of successful bioaugmentation. The results of the project are reported in three article manuscripts, included in this thesis. In manuscript I, the mineralization of 2...

  8. Bacterial mitotic machineries

    DEFF Research Database (Denmark)

    Gerdes, Kenn; Møller-Jensen, Jakob; Ebersbach, Gitte

    2004-01-01

    Here, we review recent progress that yields fundamental new insight into the molecular mechanisms behind plasmid and chromosome segregation in prokaryotic cells. In particular, we describe how prokaryotic actin homologs form mitotic machineries that segregate DNA before cell division. Thus, the Par......M protein of plasmid R1 forms F actin-like filaments that separate and move plasmid DNA from mid-cell to the cell poles. Evidence from three different laboratories indicate that the morphogenetic MreB protein may be involved in segregation of the bacterial chromosome....

  9. Changes in bacterial meningitis.

    OpenAIRE

    Carter, P E; Barclay, S M; Galloway, W H; Cole, G F

    1990-01-01

    In 1964, one of us (WHG) undertook a retrospective study of bacterial meningitis in childhood in the north east of Scotland during the period 1946-61. We have recently carried out a similar review of cases occurring during 1971-86, to compare the incidence, mortality, and bacteriological patterns. During the earlier period 285 cases occurred, a total incidence of 16.9/100,000 children per year. In the later period 274 children were affected, an annual incidence of 17.8/100,000. The overall mo...

  10. Soil bacterial flora and enzymatic activities in zinc and lead ...

    African Journals Online (AJOL)

    JTEkanem

    Abstract. Soil bacterial flora and enzymatic activities in lead and zinc contaminated soil of Ishiagu, ... the quality of the soil. The type of activities prevalent in any given environment determines the type of contamination in that area1-3. Soil and water bodies have been sinks for many ..... lead, Cadmium and Mercury in cattle.

  11. Development of the gastric morphology and fornical bacterial ...

    African Journals Online (AJOL)

    area for bacterial attachment thus facilitating a symbiotic relationship between the rodent and the gastric micro- ... stomach weight, expressed as a percentage of body weight, was used as a relative measure of gastric ... from subsequent litters were removed from their mothers at progressively earlier ages and survival was ...

  12. Determination of neutral current couplings from neutrino-induced semi-inclusive pion and inclusive reactions

    International Nuclear Information System (INIS)

    Hung, P.Q.

    1977-01-01

    It is shown that by looking at data from neutrino-induced semi-inclusive pion and inclusive reactions on isoscalar targets along, one can determine completely the neutral current couplings. Predictions for various models are also presented. (Auth.)

  13. Supporting local institutions for inclusive green growth

    NARCIS (Netherlands)

    Berkhout, Ezra; Bouma, Jetske; Terzidis, Nikolaos; Voors, Maarten

    2018-01-01

    We conduct a structured search of the academic literature that assesses the impact of development interventions that aim to build and strengthen local-level institutions to facilitate Inclusive Green Growth. Inclusive Green Growth extends the standard growth perspective to include welfare

  14. Towards an Elaborated Theory of Inclusive Development

    NARCIS (Netherlands)

    Gupta, J.; Pouw, N.R.M.; Ros-Tonen, M.A.F.

    2015-01-01

    Sustainable development often leads to strong trade-offs, mostly in favour of economic growth. Inclusive development responds by focusing mainly on the social and environmental aspects of development and on current generations. While the literature covers inclusive growth in some detail, few authors

  15. Preparation and characterization of artemether inclusion complexes ...

    African Journals Online (AJOL)

    Purpose: To investigate experimentally the inclusion of artemether into the cavity of hydroxypropyl-β-cyclodextrin and examine its effect on the solubility and dissolution rate of the drug. Methods: Inclusion complexes of artemether with hydroxypropyl-β-cyclodextrin of molar ratios 1:1, 1:2 and 1:3 were prepared using the ...

  16. Early Childhood Inclusion in the United Kingdom

    Science.gov (United States)

    Blackburn, Carolyn

    2016-01-01

    A policy-to-practice paper is presented of early childhood inclusion in England. The article aims to report the benefits of early intervention services and early childhood inclusion for children with special educational needs and disabilities (SEND), document the chronology of policy development, and discuss research evidence about…

  17. Inclusion complexation of tetrabutylammonium iodide by cyclodextrins

    Indian Academy of Sciences (India)

    Host-guest inclusion complex of an ionic solid (tetrabutyl ammonium iodide) with α- and β- cyclodextrin has been explored by various physicochemical and spectroscopic methods. Surface tension and conductivity studies indicated 1:1 stoichiometry of the inclusion complexes and ¹H NMR and FT-IR studies substantiated ...

  18. Attitudes of German Student Teachers on Inclusion

    Science.gov (United States)

    Baar, Robert

    2016-01-01

    The contribution discusses attitudes of German Teacher Training Students on Inclusion based on an empirical analysis containing three elements: Evaluation of students' written exams, results of a survey with closed as open questions and the interpretation of group discussions among students about inclusion. One can see that, though the found-out…

  19. Proving program inclusion using Hoare's logic

    NARCIS (Netherlands)

    Bergstra, J.A.; Klop, J.W.

    1984-01-01

    We explore conservative refinements of specifications. These form a quite appropriate framework for a proof theory for program inclusion based on a proof theory for program correctness. We propose two formalized proof methods for program inclusion and prove these to be sound. Both methods are

  20. Inclusive photoproduction and the triple Regge formula

    International Nuclear Information System (INIS)

    Gandsman, J.

    The inclusive reactions γp→π + or π - + anything and γn→π + or π - + anything at the photon energy of 7.5GeV were studied. The high π + /π - ratio observed in the target fragmentation region is explained by applying the triple Regge formalism. The inclusive photoproduction of the Δ ++ and rho 0 resonances is also investigated [fr

  1. Green Inclusion: Biophilia as a Necessity

    Science.gov (United States)

    Stavrianos, Alexandros

    2016-01-01

    This article argues that there is a strong link between the pedagogy for inclusion and the pedagogy of environmental education and tries to identify any benefits that could be acquired by pupils when the school system tries to use environmental educational programmes to promote inclusion. The idea of promoting a school yard into a school garden is…

  2. State Early Childhood Inclusion Self-Assessment

    Science.gov (United States)

    Whaley, Kathy T.; Cate, Debbie; Dell, Penny; Vinh, Megan; Neitzel, Jen

    2017-01-01

    This self-assessment tool provides a framework for examining key aspects of a State infrastructure that are useful for promoting high quality inclusive practices, programs and policies. The sections of the self-assessment are organized by the nine state recommendations of the "Policy Statement on Inclusion of Children with Disabilities in…

  3. Inclusive growth: Buzzword or innovation? | IDRC - International ...

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

    2017-05-03

    May 3, 2017 ... The World Economic Forum (WEF) brought great focus to this debate through its Inclusive Growth and Development Report, which was the subject of a recent event in Ottawa at the International Development Research Centre (IDRC). The report's lead author, Richard Samans, argues that an inclusive ...

  4. Social Justice Leadership and Inclusion: A Genealogy

    Science.gov (United States)

    Lewis, Katherine

    2016-01-01

    The purpose of this article is to engage in an historical analysis of research about two concepts: social justice leadership and leadership for inclusion. Recent experiences have caused me to wonder about our interpretations of justice, equity, and inclusion. Analysis of the relevant literature revealed a lack of consensus among scholars as to a…

  5. Inclusive Partnership: Enhancing Student Engagement in Geography

    Science.gov (United States)

    Moore-Cherry, Niamh; Healey, Ruth; Nicholson, Dawn T.; Andrews, Will

    2016-01-01

    Partnership is currently the focus of much work within higher education and advocated as an important process to address a range of higher education goals. In this paper, we propose the term "inclusive partnership" to conceptualise a non-selective staff-student relationship. While recognising the challenges of inclusive partnership…

  6. School Inclusion and the "Community of Practice"

    Science.gov (United States)

    Laluvein, Jackie

    2010-01-01

    "Inclusion" is not a mechanism for relocating educationally disadvantaged youngsters in mainstream rather than in special schools. Rather, inclusion implies a whole school approach to social relations and production of meaning reached through processes of negotiation between parents, teachers and children. Such an approach places equal value upon…

  7. Teachers' Attitudes towards Inclusion in High Schools

    Science.gov (United States)

    Boyle, Christopher; Topping, Keith; Jindal-Snape, Divya

    2013-01-01

    This study investigated the attitudes of secondary teachers to inclusion in schools. Three hundred and ninety-one teaching and management-level staff from 19 mainstream and 6 special schools in one local authority in Scotland completed a survey. Overall, teaching staff were pro-inclusion, conditional on adequate support and resources. There was a…

  8. Responsible Inclusion: Celebrating Diversity and Academic Excellence.

    Science.gov (United States)

    Malloy, William

    1997-01-01

    The North Carolina Partnership Training System, a federally funded technical assistance project, convened a Principals Task Force to examine inclusion practices. They identified several facilitators and barriers to inclusion and found that block scheduling, collaborative teaching, cooperative learning, interdisciplinary planning, heterogeneous…

  9. Erasmus Mundus SEN: The Inclusive Scholarship Programme?

    Science.gov (United States)

    Grinbergs, Christopher J.; Jones, Hilary

    2013-01-01

    The Erasmus Mundus MA/Mgr in Special Education Needs (EM SEN) was created as a Masters Course funded by the European Commission's Erasmus Mundus Programme (EMP) to challenge and educate students in inclusive policy and practice in education. Yet, it is debatable the extent to which this programme embodies the values of an inclusive approach,…

  10. Guided Reading in Inclusive Middle Years Classrooms

    Science.gov (United States)

    Lyons, Wanda; Thompson, Scott Anthony

    2012-01-01

    Teachers in inclusive classrooms are challenged to provide reading instruction for students with a wide range of instructional levels. This article reports on the implementation of guided reading in four middle years inclusive classrooms, the impact on student engagement and reading progress, and teacher perspectives on the guided reading…

  11. How Does Supervision Support Inclusive Teacherhood?

    Science.gov (United States)

    Alila, Sanna; Määttä, Kaarina; Uusiautti, Satu

    2016-01-01

    Supervision is a multidimensional concept and phenomenon. In this study, the advantages of supervision and its development in inclusive teacherhood was studied. Inclusive teacherhood means a teacher's professional development and the school culture's change toward participatory school for all students. The study analyzed the views of supervisors…

  12. On Graph Refutation for Relational Inclusions

    Directory of Open Access Journals (Sweden)

    Paulo A. S. Veloso

    2012-03-01

    Full Text Available We introduce a graphical refutation calculus for relational inclusions: it reduces establishing a relational inclusion to establishing that a graph constructed from it has empty extension. This sound and complete calculus is conceptually simpler and easier to use than the usual ones.

  13. Older Women's Career Development and Social Inclusion

    Science.gov (United States)

    McMahon, Mary; Bimrose, Jenny; Watson, Mark

    2010-01-01

    This paper considers women's career development and the potential contribution of career development theory, research, practice and policy in advancing a social inclusion agenda. In particular, the paper focuses on older women in the contexts of an ageing population, labour market shortages and Australia's social inclusion agenda. Supporting young…

  14. Leading under Pressure: Leadership for Social Inclusion

    Science.gov (United States)

    Muijs, Daniel; Ainscow, Mel; Dyson, Alan; Raffo, Carlo; Goldrick, Sue; Kerr, Kirstin; Lennie, Clare; Miles, Susie

    2010-01-01

    In this study we undertook to look at leadership issues specifically in relation to social inclusion, through a series of six case studies in three districts showing high levels of disadvantage. Findings indicated that schools' views on social inclusion could be typified as leaning towards three main orientations: (1) improving achievement and…

  15. Social Inclusion and Critical Consciousness in Australia

    Science.gov (United States)

    Diemer, Matthew A.; Ortega, Lilyana

    2010-01-01

    Australia's Indigenous population is excluded from a range of opportunities, experiences and amenities that facilitate wellbeing, self-determination and social inclusion. This social exclusion constrains the career development and occupational attainment of Indigenous youth, which represent key routes to societal inclusion. Critical…

  16. Implementing inclusive educational practices through partnerships ...

    African Journals Online (AJOL)

    This study reports on work in progress of a partnership between the University of Stellenbosch and three rural schools in a disadvantaged community, focused on the development of inclusive educational practices such as teaching, assessment and support in inclusive education. Recognizing the changing needs of the ...

  17. Inclusive Education in Thailand: Practices and Challenges

    Science.gov (United States)

    Vorapanya, Sermsap; Dunlap, Diane

    2014-01-01

    In 2008, Thailand passed legislation on the educational provisions for students with disabilities to mandate the implementation of inclusive education. This article provides a historical overview of special education in Thailand and the emergence of inclusive education as it moves from policy to practice. To further identify the challenges faced…

  18. Understanding "Inclusive Growth": Advancing the global agenda ...

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

    2013-03-27

    Mar 27, 2013 ... Increasingly, the concept of "inclusive growth," where the benefits of economic growth are enjoyed across societies, is being recognized and advanced by many countries. Growth is less likely to be ... Mini soap operas foster financial education and inclusion of women in Peru. Having little or no savings can ...

  19. Teachers' Misunderstanding: The Concept of Inclusive Education

    Science.gov (United States)

    Sanagi, Tomomi

    2016-01-01

    Teachers' misunderstanding the concept of inclusive education will not lead to good practices, rather make an exclusive environment for pupils with special educational needs in mainstream schools. This study clarified teachers' attitudes towards the image of inclusive education with conjoint analysis and cluster analysis. The participants for this…

  20. Towards inclusion? Developing teacher skills for communication ...

    African Journals Online (AJOL)

    USER

    Key Words: Teacher training, Communication, Deaf, Hard of hearing, inclusion. Introduction. Inclusion of children with special needs is now a dominant policy and practice worldwide. This form of education provision is being championed with a view to meeting the diverse educational needs of all children (UNESCO,. 1994.

  1. Moving Forward in Inclusive Education Research

    Science.gov (United States)

    Erten, Ozlem; Savage, Robert Samuel

    2012-01-01

    This paper aims to address conceptual and methodological challenges of doing research in the field of inclusive education and revisit school effectiveness research literature to inform future research. First, we present the rationale for inclusive education and briefly review the evolution of special needs education. Then, we discuss limitations…

  2. Inclusive Education: Teacher Perspectives from Malaysia

    Science.gov (United States)

    Bailey, Lucy; Nomanbhoy, Alefiya; Tubpun, Tida

    2015-01-01

    This paper reports on a survey of views of inclusive education expressed by nearly 300 Malaysian primary school teachers involved in remedial literacy and numeracy education under the country's Literacy and Numeracy Strategy. Overall, the views expressed were positive towards the principle of inclusion. However, despite common professional…

  3. Students' Perspectives on LGBTQ-Inclusive Curriculum

    Science.gov (United States)

    Snapp, Shannon D.; Burdge, Hilary; Licona, Adela C.; Moody, Raymond L.; Russell, Stephen T.

    2015-01-01

    Implementing curriculum that is inclusive of lesbian, gay, bisexual, transgender, queer, and questioning (LGBTQ) people has the potential to create an equitable learning environment. In order to learn more about students' experiences of LGBTQ-inclusive curriculum, 26 high school students with diverse racial/ethnic, sexual, and gender identities…

  4. Thermal gradient brine inclusion migration in salt study: gas-liquid inclusions, preliminary model

    International Nuclear Information System (INIS)

    Olander, D.R.; Machiels, A.J.

    1979-10-01

    Natural salt deposits contain small cubical inclusions of brine distributed through the salt. Temperature gradients, resulting from storing heat-generating wastes in the salt, can cause the inclusions to move through the salt. Prediction of the rate and amount of brine-inclusion migration is necessary for the evaluation of bedded or domed salts as possible media for waste repositories. Inclusions filled exclusively with liquid migrate up the temperature gradient towards the heat source. The solubility of salt in the brine inclusion increases with temperature. Consequently, salt dissolves into the inclusion across the hot surface and crystallizes out at the cold surface. Diffusion of salt within the liquid phase from the hot to the cold faces causes the inclusions to move in the opposite direction. In so doing, they change shape and eventually become rectangular parallelipipeds with a width (dimension perpendicular to the thermal gradient) much larger than the thickness (dimension in the direction of the thermal gradient). The inclusions may also contain a gas phase predominantly consisting of water vapor. These entities are termed two-phase or gas-liquid inclusions. The two-phase inclusions usually migrate down the temperature gradient away from the heat source remaining more-or-less cubical. A two-phase inclusion also forms when an all-liquid inclusion reaches the waste package; upon opening up at the salt-package interface, the brine partially evaporates and the inclusion reseals with some insoluble gas trapped inside. These gas-liquid inclusions proceed to move down the temperature gradient, in the opposite sense of the all-liquid inclusions. The gas-liquid inclusions phenomenon provides a pathway by which radionuclides leached from the wasteform by the brine can be transported away from the waste package and thus might have greater access to the biosphere

  5. Body Piercing

    Science.gov (United States)

    ... Health Medical Conditions Nutrition & Fitness Emotional Health Body Piercing Posted under Health Guides . Updated 1 August 2017. + ... medical reasons why I should not get a piercing? Yes. There are medical conditions (see the list ...

  6. The Politics of Inclusion and Empowerment

    DEFF Research Database (Denmark)

    Andersen, John; Siim, Birte

    identities. Politics of empowerment has to do with the agency and mobilisation dimension of social and political change. The title of the book "Politics of Inclusion and Empowerment" address the leitmotiv: namely to discuss plussumgame between politics of inclusion and politics of empowerment......The objective of the book is to analyse different politics of inclusion and empowerment and the different paradigms of inclusion/exclusion in order to underline the close link between politics of scoial equality and politics of recognition of ciultural difference. Politics of inclusion is thus...... theproductive/innovative linkage of politics of redistributuin and politics og resognition, whnich over a longer time span creates sustainable paths of democratic and social development, which increases the capacity to handle both conflicts about economic resources and life-chances and conflicts about...

  7. Creating Inclusive Youth Programs for LGBTQ+ Communities

    Directory of Open Access Journals (Sweden)

    Katherine E. Soule

    2017-06-01

    Full Text Available It is vital for youth to experience inclusive programming that is welcoming. Extension has a responsibility and an obligation to provide youth with programs and spaces that are inclusive of all sexes, gender identities, gender expressions, and sexual orientations. This article provides an overview of appropriate terminology, as well as steps for creating inclusive Extension spaces and programs for youth who identify as members of lesbian, gay, bisexual, transgender, or queer/questioning (LGBTQ+ communities. With a focus on urban Extension audiences, this article uses accessible language, self-reflective prompts, and supporting visual aids to share lessons learned from ongoing inclusivity trainings with Extension personnel across the nation, as well as from research activities and inclusive programming.

  8. Inclusive fitness maximization: An axiomatic approach.

    Science.gov (United States)

    Okasha, Samir; Weymark, John A; Bossert, Walter

    2014-06-07

    Kin selection theorists argue that evolution in social contexts will lead organisms to behave as if maximizing their inclusive, as opposed to personal, fitness. The inclusive fitness concept allows biologists to treat organisms as akin to rational agents seeking to maximize a utility function. Here we develop this idea and place it on a firm footing by employing a standard decision-theoretic methodology. We show how the principle of inclusive fitness maximization and a related principle of quasi-inclusive fitness maximization can be derived from axioms on an individual׳s 'as if preferences' (binary choices) for the case in which phenotypic effects are additive. Our results help integrate evolutionary theory and rational choice theory, help draw out the behavioural implications of inclusive fitness maximization, and point to a possible way in which evolution could lead organisms to implement it. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Diffusion of nanosized sodium inclusions in platinum

    International Nuclear Information System (INIS)

    Poulsen, J.R.; Horsewell, A.; Eldrup, M.; Johnson, E.; Johansen, A.

    1994-01-01

    Na inclusions with diameters in the range from 2 nm to 15 nm have been made by ion implantation of Na into 70 nm thick single-crystalline Pt foils followed by annealing. The structure of solid inclusions and the diffusion of molten inclusions have been studied by transmission electron microscopy. At room temperature the inclusions are faceted and crystalline with a BCC structure and they are aligned topotactically with the Pt (FCC) matrix. The diffusion of inclusions in the liquid state was investigated by annealing at temperatures of 1227 K, 1432 K and 1534 K. The results are used to propose a method to produce sources for positron annihilation spectroscopy at high temperatures. (author)

  10. Statistical model for high energy inclusive processes

    International Nuclear Information System (INIS)

    Pomorisac, B.

    1980-01-01

    We propose a statistical model of inclusive processes. The model is an extension of the model proposed by Salapino and Sugar for the inclusive distributions in rapidity. The model is defined in terms of a random variable on the full phase space of the produced particles and in terms of a Lorentz-invariant probability distribution. We suggest that the Lorentz invariance is broken spontaneously, this may describe the observed anisotropy of the inclusive distributions. Based on this model we calculate the distribution in transverse momentum. An explicit calculation is given of the one-particle inclusive cross sections and the two-particle correlation. The results give a fair representation of the shape of one-particle inclusive cross sections, and positive correlation for the particles emitted. The relevance of our results to experiments is discussed

  11. Inclusive practices in teacher training in Mexico

    Directory of Open Access Journals (Sweden)

    Vasthi Jocabed Flores Barrera

    2017-06-01

    Full Text Available Inclusive practices include diversity as a resource that favors teaching-learning processes in the classroom, although they focus on the most vulnerable people by offering them higher-quality education. This descriptive study sought to identify the inclusive practices of teachers who train teachers in an Escuela Normal (teachers’ college in Mexico. Eight teachers and 247 students participated in the study; the Guidelines for the Evaluation of Inclusive Practices in the Classroom (observation formats and students, the Learning Strategies Questionnaire, and semi-structured interviews were used. The results suggest that, although the general perception is that participating teachers have high inclusive practices, they need greater support in the physical conditions of the classroom, methodology and teacher-student relationship. It was also identified the need for a refresher program for teachers to conceptually enrich the teaching staff and encourage the implementation of inclusive education within the Escuela Normal.

  12. Inclusion in High-Achieving Singapore: Challenges of Building an Inclusive Society in Policy and Practice

    Science.gov (United States)

    Walker, Zachary; Musti-Rao, Shobana

    2016-01-01

    Building an inclusive society in which all people can participate effectively and live together requires understanding inclusive education and its impact on the social order. As countries of different regions face the vast array of challenges unique to their educational systems, it becomes apparent that inclusive societies are intricately tied to…

  13. More Policies, Greater Inclusion? Exploring the Contradictions of New Labour Inclusive Education Policy

    Science.gov (United States)

    Roulstone, Alan; Prideaux, Simon

    2008-01-01

    The era of New Labour government has witnessed unprecedented growth in inclusive education policies. There is, however, limited evidence that policies have increased disabled children's inclusion. This article explores reasons for this contradiction. Drawing on sociological insights, it is argued that New Labour policies on inclusive education…

  14. Alaska Inclusion Training Modules: Building Inclusive Classrooms and Schools. Modules 1-3. Second Edition.

    Science.gov (United States)

    Ryan, Susan, Ed.; Kurka, Diana, Ed.

    This document presents three inservice training modules which focus on the inclusion of students with disabilities in regular education settings in Alaska schools. Module 1, "Setting our Sights on Inclusion," by Diana Kurka, is a three-hour inservice program which provides an introduction to inclusion via sections on: legal and…

  15. Full Inclusion: The Benefits and Disadvantages of Inclusive Schooling. An Overview

    Science.gov (United States)

    McCarty, Kristine

    2006-01-01

    An overview of debated issues related to the benefits and disadvantages of inclusive schooling. The beliefs of districts, school personnel and parents are widely differing as related to the placement of students with special needs in inclusive settings. The examples of current federal laws help to shape the debate of full inclusion in schools…

  16. Beyond Physical Inclusion: Teaching Skills in the Community to Enhance Social Inclusion

    Science.gov (United States)

    Hall, Carmen L.

    2017-01-01

    Along with the deinstitutionalization movement, supports for persons with Intellectual Disabilities (ID) have shifted to promotion of person-centered supports inclusive in the community. Although successes have occurred regarding physical inclusion, skill building and social inclusion have not fared as well for those with more significant…

  17. Nuclear Glycogen Inclusions in Canine Parietal Cells.

    Science.gov (United States)

    Silvestri, S; Lepri, E; Dall'Aglio, C; Marchesi, M C; Vitellozzi, G

    2017-05-01

    Nuclear glycogen inclusions occur infrequently in pathologic conditions but also in normal human and animal tissues. Their function or significance is unclear. To the best of the authors' knowledge, no reports of nuclear glycogen inclusions in canine parietal cells exist. After initial observations of nuclear inclusions/pseudoinclusions during routine histopathology, the authors retrospectively examined samples of gastric mucosa from dogs presenting with gastrointestinal signs for the presence of intranuclear inclusions/pseudoinclusions and determined their composition using histologic and electron-microscopic methods. In 24 of 108 cases (22%), the authors observed various numbers of intranuclear inclusions/pseudoinclusions within scattered parietal cells. Nuclei were characterized by marked karyomegaly and chromatin margination around a central optically empty or slightly eosinophilic area. The intranuclear inclusions/pseudoinclusions stained positive with periodic acid-Schiff (PAS) and were diastase sensitive, consistent with glycogen. Several PAS-positive/diastase-sensitive sections were further examined by transmission electron microscopy, also using periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) staining to identify polysaccharides. Ultrastructurally, the nuclear inclusions were composed of electron-dense particles that were not membrane bound, without evidence of nuclear membrane invaginations or cytoplasmic organelles in the nuclei, and positive staining with PA-TCH-SP, confirming a glycogen composition. No cytoplasmic glycogen deposits were observed, suggesting that the intranuclear glycogen inclusions were probably synthesized in loco. Nuclear glycogen inclusions were not associated with gastritis or colonization by Helicobacter-like organisms ( P > .05). Our findings suggest that nuclear glycogen inclusions in canine parietal cells could be an incidental finding. Nevertheless, since nuclear glycogen is present in several pathologic

  18. Bacterial microbiome of lungs in COPD

    Directory of Open Access Journals (Sweden)

    Sze MA

    2014-02-01

    Full Text Available Marc A Sze,1 James C Hogg,2 Don D Sin1 1Department of Medicine, 2Department of Pathology and Laboratory Medicine, The James Hogg Research Centre, Providence Heart-Lung Institute, St Paul's Hospital, University of British Columbia, Vancouver, BC, Canada Abstract: Chronic obstructive pulmonary disease (COPD is currently the third leading cause of death in the world. Although smoking is the main risk factor for this disease, only a minority of smokers develop COPD. Why this happens is largely unknown. Recent discoveries by the human microbiome project have shed new light on the importance and richness of the bacterial microbiota at different body sites in human beings. The microbiota plays a particularly important role in the development and functional integrity of the immune system. Shifts or perturbations in the microbiota can lead to disease. COPD is in part mediated by dysregulated immune responses to cigarette smoke and other environmental insults. Although traditionally the lung has been viewed as a sterile organ, by using highly sensitive genomic techniques, recent reports have identified diverse bacterial communities in the human lung that may change in COPD. This review summarizes the current knowledge concerning the lung microbiota in COPD and its potential implications for pathogenesis of the disease. Keywords: chronic obstructive pulmonary disease, bacterial microbiome, lungs

  19. Corruption of innate immunity by bacterial proteases.

    Science.gov (United States)

    Potempa, Jan; Pike, Robert N

    2009-01-01

    The innate immune system of the human body has developed numerous mechanisms to control endogenous and exogenous bacteria and thus prevent infections by these microorganisms. These mechanisms range from physical barriers such as the skin or mucosal epithelium to a sophisticated array of molecules and cells that function to suppress or prevent bacterial infection. Many bacteria express a variety of proteases, ranging from non-specific and powerful enzymes that degrade many proteins involved in innate immunity to proteases that are extremely precise and specific in their mode of action. Here we have assembled a comprehensive picture of how bacterial proteases affect the host's innate immune system to gain advantage and cause infection. This picture is far from being complete since the numbers of mechanisms utilized are as astonishing as they are diverse, ranging from degradation of molecules vital to innate immune mechanisms to subversion of the mechanisms to allow the bacterium to hide from the system or take advantage of it. It is vital that such mechanisms are elucidated to allow strategies to be developed to aid the innate immune system in controlling bacterial infections.

  20. Corruption of Innate Immunity by Bacterial Proteases

    Science.gov (United States)

    Potempa, Jan; Pike, Robert N.

    2009-01-01

    The innate immune system of the human body has developed numerous mechanisms to control endogenous and exogenous bacteria and thus prevent infections by these microorganisms. These mechanisms range from physical barriers such as the skin or mucosal epithelium to a sophisticated array of molecules and cells that function to suppress or prevent bacterial infection. Many bacteria express a variety of proteases, ranging from non-specific and powerful enzymes that degrade many proteins involved in innate immunity to proteases that are extremely precise and specific in their mode of action. Here we have assembled a comprehensive picture of how bacterial proteases affect the host’s innate immune system to gain advantage and cause infection. This picture is far from being complete since the numbers of mechanisms utilized are as astonishing as they are diverse, ranging from degradation of molecules vital to innate immune mechanisms to subversion of the mechanisms to allow the bacterium to hide from the system or take advantage of it. It is vital that such mechanisms are elucidated to allow strategies to be developed to aid the innate immune system in controlling bacterial infections. PMID:19756242