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Sample records for bacterial inclusion body

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

    SciTech Connect

    Gonzalez-Montalban, Nuria; Villaverde, Antonio; Aris, Anna; E-mail: Anna.Aris@irta.es

    2007-04-13

    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 {beta}-galactosidase fusion protein are clearly toxic for mammalian cells but the {beta}-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.

  2. Isolation of biologically active nanomaterial (inclusion bodies) from bacterial cells

    PubMed Central

    2010-01-01

    Background In recent years bacterial inclusion bodies (IBs) were recognised as highly pure deposits of active proteins inside bacterial cells. Such active nanoparticles are very interesting for further downstream protein isolation, as well as for many other applications in nanomedicine, cosmetic, chemical and pharmaceutical industry. To prepare large quantities of a high quality product, the whole bioprocess has to be optimised. This includes not only the cultivation of the bacterial culture, but also the isolation step itself, which can be of critical importance for the production process. To determine the most appropriate method for the isolation of biologically active nanoparticles, three methods for bacterial cell disruption were analyzed. Results In this study, enzymatic lysis and two mechanical methods, high-pressure homogenization and sonication, were compared. During enzymatic lysis the enzyme lysozyme was found to attach to the surface of IBs, and it could not be removed by simple washing. As this represents an additional impurity in the engineered nanoparticles, we concluded that enzymatic lysis is not the most suitable method for IBs isolation. During sonication proteins are released (lost) from the surface of IBs and thus the surface of IBs appears more porous when compared to the other two methods. We also found that the acoustic output power needed to isolate the IBs from bacterial cells actually damages proteins structures, thereby causing a reduction in biological activity. High-pressure homogenization also caused some damage to IBs, however the protein loss from the IBs was negligible. Furthermore, homogenization had no side-effects on protein biological activity. Conclusions The study shows that among the three methods tested, homogenization is the most appropriate method for the isolation of active nanoparticles from bacterial cells. PMID:20831775

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed Central

    Vallejo, Luis Felipe; Rinas, Ursula

    2004-01-01

    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. PMID:15345063

  5. Inclusion Body Myositis

    MedlinePlus

    ... What is Inclusion Body Myositis? Inclusion body myositis (IBM) is one of a group of muscle diseases ... muscle weakness. The onset of muscle weakness in IBM is generally gradual (over months or years) and ...

  6. Investigation of the phase morphology of bacterial PHA inclusion bodies by contrast variation SANS

    NASA Astrophysics Data System (ADS)

    Russell, R. A.; Holden, P. J.; Garvey, C. J.; Wilde, K. L.; Hammerton, K. M.; Foster, L. J.

    2006-11-01

    Under growth-limiting conditions, many bacteria are able to metabolise excess organic acids into polyhydroxyalkanoates (PHA) and store these polymers as intracellular inclusions until the return of favourable conditions. Various models have been proposed for the macromolecular organisation of the boundary layer surrounding the polymer, and contrast-variation small-angle neutron scattering (SANS) was used to study its organisation. Inclusions formed by Pseudomonas oleovorans under hydrogenating conditions showed lowest scattering intensity at ca. 20% D 2O. The inclusions consist of protein and membrane lipids in the boundary layer and polyhydroxyoctanoate (lipid) in the inclusion body. At 20% D 2O the contributions of lipids were contrast matched with the solvent, indicating that lipids contributed the bulk of the scattering intensity observed at other D 2O/H 2O ratios. These results are inconsistent with a model of the boundary layer which proposed outer and inner layers of crystalline protein lattice sandwiching a membrane lipid membrane layer [E.S. Stuart, R.W. Lenz, R.C. Fuller, Can J Microbiol 41(Suppl 1) (1995) 84-93], and is more consistent with a model consisting of a lipid monolayer containing embedded proteins [U. Pieper-furst, M.H. Madkour, F. Mayer, A. Steinbuchel, J. Bacteriol. 176 (1994) 4328-4337.] By altering the H/D content of the precursors, we were able to collect SANS data from preparations of both deuterated and H/D copolymer inclusions, where initial PHA produced was hydrogenated followed by deuteration. Deuterated inclusions showed minimum intensity above 90% D 2O/H 2O whereas the sequentially produced copolymer (assumed to be in a core/shell arrangement) displayed minimum scattering some 20% lower, which is consistent with the increased hydrogenation of the boundary layer expected from its synthesis during supply of hydrogenated followed by deuterated precursors.

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

    PubMed Central

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

    2012-01-01

    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. PMID:22512648

  8. Inclusion Body Myositis

    PubMed Central

    Dimachkie, Mazen M.; Barohn, Richard J.

    2012-01-01

    The idiopathic inflammatory myopathies are a group of rare disorders that share many similarities. These include dermatomyositis (DM), polymyositis (PM), necrotizing myopathy (NM), and sporadic inclusion body myositis (IBM). Inclusion body myositis is the most common idiopathic inflammatory myopathy after age 50 and it presents with chronic proximal leg and distal arm asymmetric mucle weakness. Despite similarities with PM, it is likely that IBM is primarily a degenerative disorder rather than an inflammatory muscle disease. Inclusion body myositis is associated with a modest degree of creatine kinase (CK) elevation and an abnormal electromyogram demonstrating an irritative myopathy with some chronicity. The muscle histopathology demonstrates inflammatory exudates surrounding and invading nonnecrotic muscle fibers often times accompanied by rimmed vacuoles. In this chapter, we review sporadic IBM. We also examine past, essentially negative, clinical trials in IBM and review ongoing clinical trials. For further details on DM, PM, and NM, the reader is referred to the idiopathic inflammatory myopathies chapter. PMID:23117948

  9. Inclusion-Body Myositis: Diagnosis

    MedlinePlus

    ... How to Get Involved Donate Inclusion-Body Myositis (IBM) Share print email share facebook twitter google plus ... Causes/Inheritance Medical Management Research Inclusion-Body Myositis (IBM) News Gene Therapy Success in IBM- A Quest ...

  10. [Inclusion-body myositis].

    PubMed

    Benveniste, O

    2014-07-01

    Sporadic inclusion-body myositis (sIBM) presents in average at the sixth decade of life and affects three men for one woman. It is a non-lethal, slowly progressive but disabling disease. Except the striated muscles, no other organs (such as the interstitial lung) are involved. The phenotype of this myopathy is particular since it involves the axial muscles (camptocormia, swallowing dysfunction) and limb girdle (notably the quadriceps) but also the distal muscles (in particular the fingers' and wrists' flexors) in a bilateral but non-symmetrical manner. The clinical presentation is then very suggestive of the diagnosis, which remains to be proven by a muscle biopsy. Histological features defining the diagnosis associate endomysial inflammatory infiltrates with frequent invaded fibres (the myositis) and amyloid deposits generally accompanying rimmed vacuoles (the inclusions). There is still today a debate to know if this disease is at its beginning a degenerative or an auto-immune condition. Nonetheless, usual immunosuppressive drugs (corticosteroids, azathioprine, methotrexate) or polyvalent immunoglobulines remain ineffective and even may worsen the handicap. Some controlled randomized trials will soon be launched for this condition, but for now, the best therapeutic approach to slow down the rapidity of progression of the disease is to maintain muscle exercise with the help of the physiotherapists. PMID:24128435

  11. One-pot refolding of core histones from bacterial inclusion bodies allows rapid reconstitution of histone octamer.

    PubMed

    Lee, Young-Tae; Gibbons, Garrett; Lee, Shirley Y; Nikolovska-Coleska, Zaneta; Dou, Yali

    2015-06-01

    We report an optimized method to purify and reconstitute histone octamer, which utilizes high expression of histones in inclusion bodies but eliminates the time consuming steps of individual histone purification. In the newly modified protocol, Xenopus laevis H2A, H2B, H3, and H4 are expressed individually into inclusion bodies of bacteria, which are subsequently mixed together and denatured in 8M guanidine hydrochloride. Histones are refolded and reconstituted into soluble octamer by dialysis against 2M NaCl, and metal-affinity purified through an N-terminal polyhistidine-tag added on the H2A. After cleavage of the polyhistidine-tag, histone octamer is further purified by size exclusion chromatography. We show that the nucleosomes reconstituted using the purified histone octamer above are fully functional. They serve as effective substrates for the histone methyltransferases DOT1L and MLL1. Small angle X-ray scattering further confirms that the reconstituted nucleosomes have correct structural integration of histone octamer and DNA as observed in the X-ray crystal structure. Our new protocol enables rapid reconstitution of histone octamer with an optimal yield. We expect this simplified approach to facilitate research using recombinant nucleosomes in vitro. PMID:25687285

  12. Inclusion Body Myositis

    PubMed Central

    Barohn, Richard J.

    2014-01-01

    The idiopathic inflammatory myopathies (IIM) are a heterogenous group of rare disorders that share many similarities. In addition to sporadic inclusion body myositis (IBM), these include dematomyositis (DM), polymyositis (PM), and autoimmune necrotizing myopathy (NM). For discussion of later three disorders, the reader is referred to the IIM review in this issue. IBM is the most common IIM after age 50. It typically presents with chronic insidious proximal leg and/or distal arm asymmetric muscle weakness leading to recurrent falls and loss of dexterity. Creatine kinase (CK) is up to 15 times elevated in IBM and needle electromyograhy (EMG) mostly shows a chronic irritative myopathy. Muscle histopathology demonstrates endomysial inflammatory exudates surrounding and invading non-necrotic muscle fibers often times accompanied by rimmed vacuoles and protein deposits. Despite inflammatory muscle pathology suggesting similarity with PM, it likely that IBM is has a prominent degenerative component as supported by refractoriness to immunosuppressive therapy. We review the evolution of our knowledge in IBM with emphasis on recent developments in the field and discuss ongoing clinical trials. PMID:25037082

  13. Genetics Home Reference: inclusion body myopathy 2

    MedlinePlus

    ... Conditions inclusion body myopathy 2 inclusion body myopathy 2 Enable Javascript to view the expand/collapse boxes. ... Open All Close All Description Inclusion body myopathy 2 is a condition that primarily affects skeletal muscles , ...

  14. Inclusion body hepatitis in kestrels (Falco sparverius)

    USGS Publications Warehouse

    Sileo, L.; Franson, J.C.; Graham, D.L.; Domermuth, C.H.; Rattner, B.A.; Pattee, O.H.

    1982-01-01

    Inclusion body disease of suspected adenovirus etiology was the apparent cause of death of 9 captive kestrels (Falco sparverius). Cloacal hemorrhage was the only prominent gross lesion; disseminated hepatocellular necrosis and intranuclear inclusion bodies were evident microscopically. Attempts to reproduce the disease, and to propagate and serologically characterize the agent were unsuccessful.

  15. Recovery of active anti TNF-α ScFv through matrix-assisted refolding of bacterial inclusion bodies using CIM monolithic support.

    PubMed

    Sushma, Krishnan; Bilgimol, Chuvappumkal Joseph; Vijayalakshmi, Mookambeswaran A; Satheeshkumar, Padikara Kutty

    2012-04-01

    Anti TNF-α molecules are important as therapeutic agents for many of the autoimmune diseases in chronic stage. Here we report the expression and purification of a recombinant single chain variable fragment (ScFv) specific to TNF-α from inclusion bodies. In contrast to the conventional on column refolding using the soft gel supports, an efficient methodology using monolithic matrix has been employed. Nickel (II) coupled to convective interaction media (CIM) support was utilized for this purpose with 6M guanidine hydrochloride (GuHCl) as the chaotropic agent. The protein purified after solubilization and refolding proved to be biologically active with an IC₅₀ value of 15 μg. To the best of our knowledge, this is the first report showing the application of methacrylate based chromatographic supports for matrix-assisted refolding and purification of Escherichia coli inclusion bodies. The results are promising to elaborate the methodology further to exploit the potential positive features of monoliths in protein refolding science. PMID:22386363

  16. Epidemic adenovirus inclusions body hepatitis of the chicken in Australia.

    PubMed

    Wells, R J; Westbury, H A; Harrigan, K E; Coleman, G D; Beilharz, R G

    1977-12-01

    The occurrence of inclusion body hepatitis (IBH) on 11 properties in Australia is described. The 6 Victorian cases occurred between 1967 and 1973 and were identified as incidental findings accompanying other diseases or occurred as ill-defined, minor economic disease episodes. In well-managed flocks the extra mortality attributable to IBH was as low as 1.8% which represented a 50% increase in total mortality. The 1973 epidemic in New South Wales on 5 related farms resulted in flock mortality of 8.4% to 32.3% and an average mortality of 19.5% of 814,000 birds. For uninfected flocks of 307,000 birds the average mortality was 8.5%. Higher mortality in male flocks was statistically highly significant as was lowered efficiency of feed conversion. Growth rate in both sexes was depressed by about 0.15 kg in 70 days and heavy downgrading at processing occurred. Secondary infections were important. The cardinal features of IBH were fatal necrotising hepatitis with Cowdry type-A intranuclear inclusion body formation; secondary bacterial infections were also important causes of loss. PMID:207252

  17. Diagnostic criteria for inclusion body myositis.

    PubMed

    Hilton-Jones, D; Brady, S

    2016-07-01

    Inclusion body myositis (IBM) was first identified as a specific disorder about 40 years ago and is now recognized to be the most frequently presenting primary myopathy in middle age and beyond. Initial characterization was based on the observation of specific pathological features distinguishing it from polymyositis. It was soon appreciated that there were also distinguishing clinical features. The earliest diagnostic criteria were heavily biased towards pathological features, but over time revised criteria have given increasing importance to certain clinical features. Until the specific cause of IBM is determined, and the basic pathogenetic mechanisms are better understood, there can be no diagnostic gold-standard against which to compare the sensitivity and specificity of any proposed diagnostic criteria, but such criteria are essential to ensure that patients entering clinical, epidemiological, genetic, pathological or therapeutic studies represent a homogeneous population. It is likely that any currently accepted diagnostic criteria will, once a gold-standard is eventually established, be shown to have 'missed' patients with atypical features, but that has to be accepted to make certain that current studies are not contaminated by patients who do not have IBM. In other words, in everyday clinical practice there will be the occasional patient who an experienced myologist strongly suspects has IBM, but does not meet current criteria - the criteria lack sensitivity. But if the criteria are so broad as to include all such atypical cases, they would be likely to include patients who do not in fact have IBM - they would lack specificity. The sensitivity and specificity of existing criteria have been reviewed recently, in so far as it is possible to do so, and found to have high specificity but variable sensitivity. PMID:27027255

  18. Junction problems for thin inclusions in elastic bodies

    NASA Astrophysics Data System (ADS)

    Khludnev, A. M.

    2016-06-01

    Equilibrium problems for a 2D elastic bodies with thin Euler-Bernoulli and Timoshenko elastic inclusions are considered. It is assumed that inclusions have a joint point, and a junction problem for these inclusions is analyzed. Existence of solutions is proved, and different equivalent formulations of problems are discussed. In particular, junction conditions at the joint point are found. A delamination of the elastic inclusions is also assumed. In this case, inequality type boundary conditions are imposed at the crack faces to prevent a mutual penetration between crack faces. A convergence to infinity of a rigidity parameter of the elastic inclusions is investigated. Limit problems are analyzed.

  19. Inclusion bodies and purification of proteins in biologically active forms.

    PubMed

    Mukhopadhyay, A

    1997-01-01

    Even though recombinant DNA technology has made possible the production of valuable therapeutic proteins, its accumulation in the host cell as inclusion body poses serious problems in the recovery of functionally active proteins. In the last twenty years, alternative techniques have been evolved to purify biologically active proteins from inclusion bodies. Most of these remain only as inventions and very few are commercially exploited. This review summarizes the developments in isolation, refolding and purification of proteins from inclusion bodies that could be used for vaccine and non-vaccine applications. The second section involves a discussion on inclusion bodies, how they are formed, and their physicochemical properties. In vivo protein folding in Escherichia coli and kinetics of in vitro protein folding are the subjects of the third and fourth sections respectively. The next section covers the recovery of bioactive protein from inclusion bodies: it includes isolation of inclusion body from host cell debris, purification in denatured state alternate refolding techniques, and final purification of active molecules. Since purity and safety are two important issues in therapeutic grade proteins, the following three sections are devoted to immunological and biological characterization of biomolecules, nature, and type of impurities normally encountered, and their detection. Lastly, two case studies are discussed to demonstrate the sequence of process steps involved. PMID:8939059

  20. Refolding of proteins from inclusion bodies: rational design and recipes.

    PubMed

    Basu, Anindya; Li, Xiang; Leong, Susanna Su Jan

    2011-10-01

    The need to develop protein biomanufacturing platforms that can deliver proteins quickly and cost-effectively is ever more pressing. The rapid rate at which genomes can now be sequenced demands efficient protein production platforms for gene function identification. There is a continued need for the biotech industry to deliver new and more effective protein-based drugs to address new diseases. Bacterial production platforms have the advantage of high expression yields, but insoluble expression of many proteins necessitates the development of diverse and optimised refolding-based processes. Strategies employed to eliminate insoluble expression are reviewed, where it is concluded that inclusion bodies are difficult to eliminate for various reasons. Rational design of refolding systems and recipes are therefore needed to expedite production of recombinant proteins. This review article discusses efforts towards rational design of refolding systems and recipes, which can be guided by the development of refolding screening platforms that yield both qualitative and quantitative information on the progression of a given refolding process. The new opportunities presented by light scattering technologies for developing rational protein refolding buffer systems which in turn can be used to develop new process designs armed with better monitoring and controlling functionalities are discussed. The coupling of dynamic and static light scattering methodologies for incorporation into future bioprocess designs to ensure delivery of high-quality refolded proteins at faster rates is also discussed. PMID:21822901

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

    PubMed

    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. PMID:21538919

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

    PubMed

    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 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. PMID:24291262

  3. Inclusion body disease (herpesvirus infection) of falcons (IBDF).

    PubMed

    Graham, D L; Mare, C J; Ward, F P; Peckham, M C

    1975-01-01

    Inclusion body disease of falcons (IBDF) is caused by a herpesvirus. The clinical course is short, 24 to 72 hours in duration, and is characterized by mild to severe depression and weakness often accompanied by anorexia. The disease is invariably fatal. The virus has a marked affinity for the reticuloendothelial system and hepatocytes,producing focal to diffuse necrosis of infected tissues accompanied by the formation of intranuclear inclusion bodies. The virus is pathogenic for American kestrels (Falco sparverius) and great horned owls (Bubo virginianus) in which typical lesions of IBDF are reproduced. The lesions of IBDF are similar to those produced by some herpesvirus infections in other avian species. PMID:163383

  4. Emerging therapeutic options for sporadic inclusion body myositis

    PubMed Central

    Alfano, Lindsay N; Lowes, Linda P

    2015-01-01

    Sporadic inclusion body myositis is the most common inflammatory muscle disorder preferentially affecting males over the age of 40 years. Progressive muscle weakness of the finger flexors and quadriceps muscles results in loss of independence with activities of daily living and eventual wheelchair dependence. Initial signs of disease are often overlooked and can lead to mis- or delayed diagnosis. The underlying cause of disease is unknown, and disease progression appears refractory to available treatment options. This review discusses the clinical presentation of inclusion body myositis and the current efforts in diagnosis, and focuses on the current state of research for both nonpharmacological and pharmacological treatment options for this patient group. PMID:26445546

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

    PubMed

    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. PMID:27295879

  6. Insights into Muscle Degeneration from Heritable Inclusion Body Myopathies

    PubMed Central

    Krause, Sabine

    2015-01-01

    Muscle mass and function are gradually lost in age-related, degenerative neuromuscular disorders, which also reflect the clinical hallmarks of sarcopenia. The consensus definition of sarcopenia includes a condition of age-related loss of muscle mass, quality, and strength. The most common acquired muscle disease affecting adults aged over 50 years is sporadic inclusion body myositis (sIBM). Besides inflammatory effects and immune-mediated muscle injury, degenerative myofiber changes are characteristic features of the disease. Although the earliest triggering events in sIBM remain elusive, a plethora of downstream mechanisms are implicated in the pathophysiology of muscle wasting. Although it remains controversial whether hereditary forms of inclusion body myopathy (IBM) may be considered as degenerative sIBM disease models, partial pathophysiological aspects can mimic the much more frequent sporadic condition, in particular the occurrence of inclusion bodies in skeletal muscle. Various clinical aspects in genetically determined skeletal muscle disorders reflect age-related alterations observed in sarcopenia. Several intriguing clues from monogenic defects in heritable IBMs contributing to the molecular basis of muscle loss will be discussed with special emphasis on inclusion body myopathy with Paget’s disease of bone and frontotemporal dementia (IBMPFD) and GNE myopathy. Finally, also the recently identified dominant multisystem proteinopathy will be considered, which may rarely present as IBM. PMID:25729363

  7. Laing distal myopathy pathologically resembling inclusion body myositis

    PubMed Central

    Roda, Ricardo H; Schindler, Alice B; Blackstone, Craig; Mammen, Andrew L; Corse, Andrea M; Lloyd, Thomas E

    2014-01-01

    Mutations in MYH7 cause autosomal dominant Laing distal myopathy. We present a family with a previously reported deletion (c.5186_5188delAGA, p.K1729del). Muscle pathology in one family member was characterized by an inflammatory myopathy with rimmed vacuoles, increased MHC Class I expression, and perivascular and endomysial muscle inflammation comprising CD3+, CD4+, CD8+, and CD68+ inflammatory cells. Interestingly, this biopsy specimen contained TDP-43, p62, and SMI-31-positive protein aggregates typical of inclusion body myositis. These findings should alert physicians to the possibility that patients with MYH7 mutations may have muscle biopsies showing pathologic findings similar to inclusion body myositis. PMID:25574480

  8. Treatment of sporadic inclusion body myositis with bimagrumab

    PubMed Central

    Amato, Anthony A.; Sivakumar, Kumaraswamy; Goyal, Namita; David, William S.; Salajegheh, Mohammad; Praestgaard, Jens; Lach-Trifilieff, Estelle; Trendelenburg, Anne-Ulrike; Laurent, Didier; Glass, David J.; Roubenoff, Ronenn

    2014-01-01

    Objective: To study activin signaling and its blockade in sporadic inclusion body myositis (sIBM) through translational studies and a randomized controlled trial. Methods: We measured transforming growth factor β signaling by SMAD2/3 phosphorylation in muscle biopsies of 50 patients with neuromuscular disease (17 with sIBM). We tested inhibition of activin receptors IIA and IIB (ActRII) in 14 patients with sIBM using one dose of bimagrumab (n = 11) or placebo (n = 3). The primary outcome was the change in right thigh muscle volume by MRI at 8 weeks. Lean body mass, strength, and function were secondary outcomes. Twelve of the patients (10 bimagrumab, 2 placebo) participated in a subsequent 16-week observation phase. Results: Muscle SMAD2/3 phosphorylation was higher in sIBM than in other muscle diseases studied (p = 0.003). Eight weeks after dosing, the bimagrumab-treated patients increased thigh muscle volume (right leg +6.5% compared with placebo, p = 0.024; left leg +7.6%, p = 0.009) and lean body mass (+5.7% compared with placebo, p = 0.014). Subsequently, bimagrumab-treated patients had improved 6-minute walking distance, which peaked at 16 weeks (+14.6%, p = 0.008) compared with placebo. There were no serious adverse events; the main adverse events with bimagrumab were mild acne and transient involuntary muscle contractions. Conclusions: Transforming growth factor β superfamily signaling, at least through ActRII, is implicated in the pathophysiology of sIBM. Inhibition of ActRII increased muscle mass and function in this pilot trial, offering a potential novel treatment of sIBM. Classification of evidence: This study provides Class I evidence that for patients with inclusion body myositis, bimagrumab increases thigh muscle volume at 8 weeks. PMID:25381300

  9. Inclusion body myositis associated with Sjögren's syndrome.

    PubMed

    Misterska-Skóra, Maria; Sebastian, Agata; Dzięgiel, Piotr; Sebastian, Maciej; Wiland, Piotr

    2013-12-01

    Inclusion body myositis (IBM) belongs to the group of idiopathic inflammatory myopathies. It is a poorly understood disease, which affects skeletal muscles. IBM usually occurs as an isolated condition, but in some cases, it may be associated with another autoimmune disorder, Sjögren's syndrome. We report a case of a 47-year-old woman with headaches, symptoms of trigeminal neuralgia, progressive weakness in muscles of the upper and lower extremities and symptoms of dry eyes and mouth. On admission, creatine kinase level was increased to 6,956 IU/mL and lactate dehydrogenase (LDH) to 1,011 U/L in the serum. The increase in inflammatory factor (CRP, ESR) levels was not found. The diagnosis of inclusion body myositis associated with Sjögren's syndrome was established on the basis of clinical picture and diagnostic tests. In this therapy, methotrexate and methylprednisolone were administered. The considerable improved muscle strength in the upper and lower extremities, improved speech and swallowing, disappearance of headache and reduction in CPK and LDH levels were found 8 months after establishing the diagnosis. Treatment with methotrexate and methylprednisolone improved the clinical symptoms and quality of life of this patient and may offer a therapeutic option for some patients with IBM and concomitant Sjögren's syndrome. PMID:23233115

  10. Bacterial communities in the fruit bodies of ground basidiomycetes

    NASA Astrophysics Data System (ADS)

    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.

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

    SciTech Connect

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

    2011-01-14

    Research highlights: {yields} Distinct inclusion bodies are developed by inhibition of UPP and ALP. {yields} The inclusion bodies differ in morphology, localization and formation process. {yields} The inclusion bodies are distinguishable by the localization of TSC2. {yields} 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.

  12. Activation of the Unfolded Protein Response in Sporadic Inclusion Body Myositis But Not in Hereditary GNE Inclusion Body Myopathy

    PubMed Central

    Nogalska, Anna; D’Agostino, Carla; Engel, W. King; Cacciottolo, Mafalda; Asada, Shinichi; Mori, Kazutoshi; Askanas, Valerie

    2015-01-01

    Muscle fibers in patients with sporadic inclusion-body myositis (s-IBM), the most common age-associated myopathy, are characterized by autophagic vacuoles and accumulation of ubiquitinated and congophilic multiprotein aggregates that contain amyloid-β and phosphorylated tau. Muscle fibers of autosomal-recessive hereditary inclusion-body myopathy due to the GNE mutation (GNE-h-IBM) display similar pathologic features, except with less pronounced congophilia. Accumulation of unfolded/misfolded proteins inside the ER lumen leads to ER stress, which elicits the unfolded protein response (UPR) as a protective mechanism. Here we demonstrate for the first time that UPR is activated in s-IBM muscle biopsies, since there was a) increased ATF4 protein and increased mRNA of its target CHOP, b) cleavage of the ATF6 and increased mRNA of its target GRP78, and c) an increase of the spliced form of XBP-1 and increased mRNA of EDEM, target of heterodimer of cleaved ATF6 and spliced XBP-1. In contrast, we did not find similar evidence of the UPR induction in GNE-h-IBM patient muscle, suggesting that different intracellular mechanisms might lead to the similar pathological phenotypes. Interestingly, cultured GNE-h-IBM muscle fibers had a robust UPR response to experimental ER stress stimuli, suggesting that the GNE mutation per se is not responsible for the lack of UPR in GNE-h-IBM biopsied muscle. PMID:25978849

  13. Interconnected Cavernous Structure of Bacterial Fruiting Bodies

    PubMed Central

    Harvey, Cameron W.; Du, Huijing; Xu, Zhiliang; Kaiser, Dale; Aranson, Igor; Alber, Mark

    2012-01-01

    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. PMID:23300427

  14. Sporadic inclusion body myositis: the genetic contributions to the pathogenesis

    PubMed Central

    2014-01-01

    Sporadic inclusion body myositis (sIBM) is the commonest idiopathic inflammatory muscle disease in people over 50 years old. It is characterized by slowly progressive muscle weakness and atrophy, with typical pathological changes of inflammation, degeneration and mitochondrial abnormality in affected muscle fibres. The cause(s) of sIBM are still unknown, but are considered complex, with the contribution of multiple factors such as environmental triggers, ageing and genetic susceptibility. This review summarizes the current understanding of the genetic contributions to sIBM and provides some insights for future research in this mysterious disease with the advantage of the rapid development of advanced genetic technology. An international sIBM genetic study is ongoing and whole-exome sequencing will be applied in a large cohort of sIBM patients with the aim of unravelling important genetic risk factors for sIBM. PMID:24948216

  15. Chemical treatment of Escherichia coli: 3. Selective extraction of a recombinant protein from cytoplasmic inclusion bodies in intact cells.

    PubMed

    Falconer, R J; O'Neill, B K; Middelberg, A P

    1999-02-20

    In previous parts of this study we developed procedures for the high-efficiency chemical extraction of soluble and insoluble protein from intact Escherichia coli cells. Although high yields were obtained, extraction of recombinant protein directly from cytoplasmic inclusion bodies led to low product purity due to coextraction of soluble contaminants. In this work, a two-stage procedure for the selective extraction of recombinant protein at high efficiency and high purity is reported. In the first stage, inclusion-body stability is promoted by the addition of 15 mM 2-hydroxyethyldisulfide (2-HEDS), also known as oxidized beta-mercaptoethanol, to the permeabilization buffer (6 M urea + 3 mM ethylenediaminetetraacetate [EDTA]). 2-HEDS is an oxidizing agent believed to promote disulfide bond formation, rendering the inclusion body resistant to solubilization in 6 M urea. Contaminating proteins are separated from the inclusion-body fraction by centrifugation. In the second stage, disulfide bonds are readily eliminated by including reducing agent (20 mM dithiothreitol [DTT]) into the permeabilization buffer. Extraction using this selective two-stage process yielded an 81% (w/w) recovery of the recombinant protein Long-R3-IGF-I from inclusion bodies located in the cytoplasm of intact E. coli, at a purity of 46% (w/w). This was comparable to that achieved by conventional extraction (mechanical disruption followed by centrifugation and solubilization). A pilot-scale procedure was also demonstrated using a stirred reactor and diafiltration. This is the first reported study that achieves both high extraction efficiency and selectivity by the chemical treatment of cytoplasmic inclusion bodies in intact bacterial cells. PMID:9921154

  16. Acid-fast intranuclear inclusion bodies in the kidneys of mallards fed lead shot

    USGS Publications Warehouse

    Locke, L.N.; Bagley, G.E.; Irby, H.D.

    1966-01-01

    Acid-fast intranuclear inclusion bodies were found in the cells of the proximal convoluted tubules of the kidneys of mallards fed one, two, three or eight number 6 lead shot and maintained on cracked or whole corn and on grain-duck pellet diets. No acid-fast inclusion bodies were found in mallards fed one or three lead shot but maintained on a duck pellet ration. Dietary factors may be responsible for the failure of mallards fed a duck pellet ration to develop lead Inclusion bodies when treated with one or three lead shot. The authors suggest these inclusion bodies can be used as presumptive evidence for lead intoxication in mallards.

  17. Inclusion body myositis and paraproteinemia: incidence and immunopathologic correlations.

    PubMed

    Dalakas, M C; Illa, I; Gallardo, E; Juarez, C

    1997-01-01

    Serum from 70 patients with sporadic inclusion body myositis (IBM) was subjected to agarose gel immunofixation electrophoresis. The IgG extracted from 9 patients with monoclonal proteins, 3 without, and 2 control subjects and was purified, biotinylated, and applied to muscle biopsy sections for immunocytochemistry and to purified muscle protein fractions for immunoblots. Sixteen of 70 (22.8%) patients with IBM, compared with 2% of age-matched controls, had a monoclonal gammopathy characterized as IgG lambda in 9 patients, IgG kappa in 4, IgM kappa in 2, and IgA lambda in 1. The mean age of IBM patients with gammopathy was 60.6 years (range, 35-77 years), compared with 66.1 years (range, 42-80 years) of the IBM patients without gammopathy. The IgG of the patients, more often than that of the control subjects, immunostained myonuclei and recognized various muscle proteins of 35 to 145 kd. We conclude that IBM, regardless of age, is frequently associated with monoclonal gammopathies, which often recognize various muscle components, especially myonuclei, suggesting disturbed immunoregulation. PMID:9005871

  18. Dynamic Transcriptional Response of Escherichia coli to Inclusion Body Formation

    PubMed Central

    Baig, Faraz; Fernando, Lawrence P.; Salazar, Mary Alice; Powell, Rhonda R.; Bruce, Terri F.; Harcum, Sarah W.

    2014-01-01

    Escherichia coli is used intensively for recombinant protein production, but one key challenge with recombinant E. coli is the tendency of recombinant proteins to misfold and aggregate into insoluble inclusion bodies (IBs). IBs contain high concentrations of inactive recombinant protein that require recovery steps to salvage a functional recombinant protein. Currently, no universally effective method exists to prevent IB formation in recombinant E. coli. In this study, DNA microarrays were used to compare the E. coli gene expression response dynamics to soluble and insoluble recombinant protein production. As expected and previously reported, the classical heat-shock genes had increased expression due to IB formation, including protein folding chaperones and proteases. Gene expression levels for protein synthesis-related and energy-synthesis pathways were also increased. Many transmembrane transporter and corresponding catabolic pathways genes had decreased expression for substrates not present in the culture medium. Additionally, putative genes represented over one-third of the genes identified to have significant expression changes due to IB formation, indicating many important cellular responses to IB formation still need to be characterized. Interestingly, cells grown in 3% ethanol had significantly reduced gene expression responses due to IB formation. Taken together, these results indicate that IB formation is complex, stimulates the heat-shock response, increases protein and energy synthesis needs, and streamlines transport and catabolic processes, while ethanol diminished all of these responses. PMID:24338599

  19. High-throughput automated refolding screening of inclusion bodies

    PubMed Central

    Vincentelli, Renaud; Canaan, Stéphane; Campanacci, Valérie; Valencia, Christel; Maurin, Damien; Frassinetti, Frédéric; Scappucini-Calvo, Loréna; Bourne, Yves; Cambillau, Christian; Bignon, Christophe

    2004-01-01

    One of the main stumbling blocks encountered when attempting to express foreign proteins in Escherichia coli is the occurrence of amorphous aggregates of misfolded proteins, called inclusion bodies (IB). Developing efficient protein native structure recovery procedures based on IB refolding is therefore an important challenge. Unfortunately, there is no “universal” refolding buffer: Experience shows that refolding buffer composition varies from one protein to another. In addition, the methods developed so far for finding a suitable refolding buffer suffer from a number of weaknesses. These include the small number of refolding formulations, which often leads to negative results, solubility assays incompatible with high-throughput, and experiment formatting not suitable for automation. To overcome these problems, it was proposed in the present study to address some of these limitations. This resulted in the first completely automated IB refolding screening procedure to be developed using a 96-well format. The 96 refolding buffers were obtained using a fractional factorial approach. The screening procedure is potentially applicable to any nonmembrane protein, and was validated with 24 proteins in the framework of two Structural Genomics projects. The tests used for this purpose included the use of quality control methods such as circular dichroism, dynamic light scattering, and crystallogenesis. Out of the 24 proteins, 17 remained soluble in at least one of the 96 refolding buffers, 15 passed large-scale purification tests, and five gave crystals. PMID:15388864

  20. Flotillin-1 (Reggie-2) Contributes to Chlamydia pneumoniae Growth and Is Associated with Bacterial Inclusion

    PubMed Central

    Korhonen, Juha T.; Puolakkainen, Mirja; Häivälä, Reetta; Penttilä, Tuula; Haveri, Anu; Markkula, Eveliina

    2012-01-01

    Chlamydiae are obligate intracellular pathogens replicating only inside the eukaryotic host. Here, we studied the effect of human flotillin-1 protein on Chlamydia pneumoniae growth in human line (HL) and A549 epithelial cell lines. RNA interference was applied to disrupt flotillin-1-mediated endocytosis. Host-associated bacteria were detected by quantitative PCR, and C. pneumoniae growth was evaluated by inclusion counts. C. pneumoniae attachment to host cells was unaffected, but bacterial intracellular growth was attenuated in the flotillin-1-silenced cells. By using confocal microscopy, we detected flotillin-1 colocalized with the inclusion membrane protein A (IncA) in the C. pneumoniae inclusion membranes. In addition, flotillin-1 was associated with IncA in detergent-resistant membrane microdomains (DRMs) in biochemical fractioning. These results suggest that flotillin-1 localizes to the C. pneumoniae inclusion membrane and plays an important role for intracellular growth of C. pneumoniae. PMID:22215737

  1. Factors influencing inclusion body formation in the production of a fused protein in Escherichia coli.

    PubMed Central

    Strandberg, L; Enfors, S O

    1991-01-01

    Different parameters that influenced the formation of inclusion bodies in Escherichia coli during production of a fused protein consisting of protein A from Staphylococcus aureus and beta-galactosidase from E. coli were examined. The intracellular expression of the fused protein was controlled by the pR promoter and its temperature-sensitive repressor. The induction temperature, the pH of the cultivation medium, and changes in the amino acid sequence in the linker region between protein A and beta-galactosidase had a profound effect on the formation of inclusion bodies. At 42 degrees C, inclusion bodies were formed only during the first hours after induction, and thereafter all the recombinant protein that was further produced appeared in a soluble and active state. Production at 39 and 44 degrees C resulted in inclusion body formation throughout the production period with 15 to 20% of the produced recombinant protein appearing as inclusion bodies. Cultivating cells without control of pH caused inclusion body formation throughout the induction period, and inclusion body formation increased with decreasing pH, and at least part of the insoluble protein was formed from the pool of soluble fusion protein within the cell. Changes in the amino acid sequence in the linker region between the two parts of the fusion protein abolished inclusion body formation. PMID:1908208

  2. Mass Purification of Nucleopolyhedrosis Virus Inclusion Bodies in the K-Series Centrifuge

    PubMed Central

    Breillatt, J. P.; Brantley, J. N.; Mazzone, H. M.; Martignoni, M. E.; Franklin, J. E.; Anderson, N. G.

    1972-01-01

    Nucleopolyhedrosis virus inclusion bodies specific for Hemerocampa pseudotsugata, Neodiprion sertifer, Porthetria dispar, and Heliothis zea have been purified by using a continuous-sample-flow-with-isopycnic-banding centrifuge in quantities up to 6 × 1013 polyhedral inclusion bodies per day. Continuous-flow methods for S-ρ type purification have been evolved to deal with mass isolation of bioparticles. PMID:5031562

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

    ERIC Educational Resources Information Center

    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,…

  4. p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions

    PubMed Central

    Watanabe, Yoshihisa; Tatebe, Harutsugu; Taguchi, Katsutoshi; Endo, Yasuhisa; Tokuda, Takahiko; Mizuno, Toshiki; Nakagawa, Masanori; Tanaka, Masaki

    2012-01-01

    α-Synuclein is the main component of Lewy bodies, the intraneuronal inclusion bodies characteristic of Parkinson’s disease. Although α-synuclein accumulation is caused by inhibition of proteasome and autophagy-lysosome, the degradation of α-synuclein inclusions is still unknown. Formation of Lewy body-like inclusions can be replicated in cultured cells by introducing α-synuclein fibrils generated in vitro. We used this cell culture model to investigate the autophagy of α-synuclein inclusions and impaired mitochondria. The intracellular α-synuclein inclusions immediately underwent phosphorylation and ubiquitination. Simultaneously they were encircled by an adaptor protein p62/SQSTM1 and directed to the autophagy-lysosome pathway in HEK293 cell line. Most phospho-α-synuclein-positive inclusions were degraded in 24 h, however, lysosomal dysfunction with bafilomycin A1 significantly affected their clearance. Moreover, inhibition of autophagy by Atg-5 siRNA treatment reduced the incorporation of α-synuclein inclusions into LC3-positive autophagosomes. Knockdown experiments demonstrated the requirement of p62 for α-synuclein autophagy. These results demonstrate that α-synuclein inclusions are preferred targets for p62-dependent autophagy. Next, we investigated the autophagic clearance of impaired mitochondria in α-synuclein inclusion-containing cells. Impaired mitochondria were almost completely eliminated after mitochondrial uncoupling even in the presence of α-synuclein inclusions, suggesting that mitochondrial clearance is not prevented by α-synuclein inclusions in HEK293 cells. PMID:23300799

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

    PubMed

    Capitini, Claudia; Conti, Simona; Perni, Michele; Guidi, Francesca; Cascella, Roberta; De Poli, Angela; Penco, Amanda; Relini, Annalisa; Cecchi, Cristina; Chiti, Fabrizio

    2014-01-01

    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. PMID:24497973

  6. TDP-43 Inclusion Bodies Formed in Bacteria Are Structurally Amorphous, Non-Amyloid and Inherently Toxic to Neuroblastoma Cells

    PubMed Central

    Capitini, Claudia; Conti, Simona; Perni, Michele; Guidi, Francesca; Cascella, Roberta; De Poli, Angela; Penco, Amanda; Relini, Annalisa; Cecchi, Cristina; Chiti, Fabrizio

    2014-01-01

    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. PMID:24497973

  7. Valosin-containing protein immunoreactivity in tauopathies, synucleinopathies, polyglutamine diseases and intranuclear inclusion body disease.

    PubMed

    Mori, Fumiaki; Tanji, Kunikazu; Toyoshima, Yasuko; Sasaki, Hidenao; Yoshida, Mari; Kakita, Akiyoshi; Takahashi, Hitoshi; Wakabayashi, Koichi

    2013-12-01

    Valosin-containing protein (VCP) is associated with multiple cellular functions, including ubiquitin-dependent protein degradation. Mutations in VCP are known to cause inclusion body myopathy with Paget's disease and frontotemporal dementia and familial amyotrophic lateral sclerosis (fALS; ALS14), both of which are characterized by trans-activation response DNA protein 43 (TDP-43)-positive neuronal cytoplasmic and nuclear inclusions. Recently, immunoreactivity for fALS-associated proteins (TDP-43, fused in sarcoma (FUS), optineurin and ubiquilin-2) were reported to be present in cytoplasmic and nuclear inclusions in various neurodegenerative diseases. However, the extent and frequency of VCP-immunoreactive structures in these neurodegenerative diseases are uncertain. We immunohistochemically examined the brains of 72 cases with neurodegenerative diseases and five control cases. VCP immunoreactivity was present in Lewy bodies in Parkinson's disease and dementia with Lewy bodies, and neuronal nuclear inclusions in five polyglutamine diseases and intranuclear inclusion body disease, as well as in Marinesco bodies in aged control subjects. However, other neuronal and glial cytoplasmic inclusions in tauopathies and TDP-43 proteinopathies were unstained. These findings suggest that VCP may have common mechanisms in the formation or degradation of cytoplasmic and nuclear inclusions of neurons, but not of glial cells, in several neurodegenerative conditions. PMID:23782134

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

    PubMed Central

    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. PMID:25992259

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

    PubMed

    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. PMID:26150942

  10. p62-enriched inclusion bodies in macrophages protect against atherosclerosis

    PubMed Central

    Sergin, Ismail; Bhattacharya, Somashubhra; Emanuel, Roy; Esen, Emel; Stokes, Carl J.; Evans, Trent D.; Arif, Batool; Curci, John A.; Razani, Babak

    2016-01-01

    Autophagy is a catabolic cellular mechanism that degrades dysfunctional proteins and organelles. Atherosclerotic plaque formation is enhanced in mice with macrophages that cannot undergo autophagy because of a deficiency of an autophagy component such as ATG5. We showed that exposure of macrophages to atherogenic lipids led to an increase in the abundance of the autophagy chaperone p62, which colocalized with polyubiquitinated proteins in cytoplasmic inclusions. p62 accumulation was increased in ATG5-null macrophages, which had large cytoplasmic ubiquitin-positive p62 inclusions. Aortas from atherosclerotic mice and plaques from human endarterectomy samples showed increased abundance of p62 and polyubiquitinated proteins that co-localized with plaque macrophages, suggesting that p62-enriched protein aggregates were characteristic of atherosclerosis. The formation of the cytoplasmic inclusions depended on p62 because lipid-loaded p62-null macrophages accumulated polyubiquitinated proteins in a diffuse cytoplasmic pattern. The failure of these aggregates to form was associated with increased secretion of IL-1β and enhanced macrophage apoptosis, which depended on the p62 ubiquitin-binding domain and at least partly involved p62-mediated clearance of NLRP3 inflammasomes. Consistent with our in vitro observations, p62-deficient mice formed greater numbers of more complex atherosclerotic plaques, and p62 deficiency further increased atherosclerotic plaque burden in mice with a macrophage-specific ablation of ATG5. Together, these data suggested that sequestration of cytotoxic ubiquitinated proteins by p62 protects against atherogenesis, a condition in which the clearance of protein aggregates is disrupted. PMID:26732762

  11. Limit for the Survivability from Potassium Decay of Bacterial Spores in Halite Fluid Inclusions

    NASA Astrophysics Data System (ADS)

    Kminek, G.; Bada, J. L.

    2001-12-01

    Vreeland et al.1 recently claimed to have isolated and cultured a viable spore forming halotolerant bacterium from a 250 million year old brine inclusion present in a salt crystal from the Salado formation. An earlier report suggested that viable bacterial spores could be revived from samples obtained from insects entombed in 25-40 million year old Dominican amber2. On the bases of these reports, Parkes3 raised the question of whether bacterial spores under some conditions might be effectively immortal. Sporulation, induced by an adverse change in the environmental conditions, is able to stabilize the DNA primarily against hydrolytic depurination for extended periods of time4. However, the organism is still exposed to ionizing radiation from the environment. Dormant spores have a reduced sensitivity to ionizing radiation per se, but unlike active organisms are unable to repair DNA damage encountered during long-term exposure to ionizing radiation. The accumulated damage may overwhelm any repair mechanism that starts in the early stage of spore germination5. The main radionuclide in a halite fluid inclusion is 40K, which accounts for 0.0117% of natural potassium. 40K decays via beta decay to 40Ca and via electron capture to 40Ar, releasing a primary gamma-ray. About 83.3 % of the beta's emitted are in the energy range of 0.3-1.3 MeV. We assume 7 g/l for an average concentration of natural potassium in a halite fluid inclusion, which means that the amount of 40K in a 10 μ l fluid inclusion is 8.19 ng. We have chosen a 10 μ l because this volume is typical of that used to obtain chemical data and in the attempts to extract bacteria. Less than a percent of the gamma decay energy is absorbed in a fluid inclusion of 10 μ l. Thus, we will not take the gamma decay energy into account for the further discussion. Almost all the beta energy is absorbed in the fluid inclusion. The total decay energy absorbed in a time period of 250 million years is about 87 kGy. The most

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

    PubMed

    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. PMID:27466712

  13. Transthyretin Val122Ile, accumulated Abeta, and inclusion-body myositis aspects in cultured muscle.

    PubMed

    Askanas, Valerie; Engel, W King; McFerrin, Janis; Vattemi, Gaetano

    2003-07-22

    Cultured muscle fibers (CMF) from a patient with inclusion-body myositis (IBM) and cardiac amyloidosis associated with the transthyretin (TTR) Val122Ile mutation contained aspects of the IBM phenotype: vacuolation, congophilic inclusions, and clusters of immunocolocalizing amyloid beta-peptide (Abeta) and TTR accumulations. These abnormalities are never present in normal human CMF. These perturbations were greatly increased after Abeta precursor protein gene transfer. The TTR mutation may be a genetic predisposition factor for the patient's IBM. PMID:12874414

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

    PubMed

    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

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

    PubMed Central

    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

  16. Inclusion.

    ERIC Educational Resources Information Center

    Nathanson, Jeanne H., Ed.

    1992-01-01

    This theme journal issue focuses on current activities of the Office of Special Education and Rehabilitative Services which stress inclusion of students with disabilities in the mainstream. It begins with a message from the Assistant Secretary, Robert R. Davila which examines the full meaning of an "inclusive" education. Next, Barbara Buswell and…

  17. Inclusion body disease in two captive Australian pythons (Morelia spilota variegata and Morelia spilota spilota).

    PubMed

    Carlisle-Nowak, M S; Sullivan, N; Carrigan, M; Knight, C; Ryan, C; Jacobson, E R

    1998-02-01

    Two captive Australian pythons, one carpet and one diamond python, presented with signs of central nervous system dysfunction. The carpet python was agitated. Its head was tilting and it was incoordinated and had convulsions. It was treated with antibiotics and anthelmintics but was eventually euthanased after failing to respond to therapy. The diamond python had flaccid paralysis of the caudal half. It was not treated and became disoriented and died. Hepatocytes from both pythons contained irregular 2 to 10 micron eosinophilic intracytoplasmic inclusion bodies. The brain of the diamond python was not available for examination. Occasional neurones in the carpet python brain contained similar inclusion bodies and other changes suggestive of viral infection. The clinical signs and histopathological findings in both pythons were consistent with boid inclusion body disease. PMID:9578777

  18. Inclusion body myositis, muscle blood vessel and cardiac amyloidosis, and transthyretin Val122Ile allele.

    PubMed

    Askanas, V; Engel, W K; Alvarez, R B; Frangione, B; Ghiso, J; Vidal, R

    2000-04-01

    Typical of sporadic inclusion body myositis muscle biopsies are vacuolated muscle fibers containing intracellular amyloid deposits and accumulations of "Alzheimer-characteristic" proteins. There is no muscle blood vessel or cardiac amyloidosis. We report on a 70-year-old African-American man homozygous for the transthyretin Val122Ile allele who has both sporadic inclusion body myositis and cardiac amyloidosis. His unique pathological features included transthyretin immunoreactivity in prominent muscle blood vessel amyloid and congophilic amyloid deposits within vacuolated muscle fibers. PMID:10762172

  19. X-ray powder pattern analysis of cytoplasmic polyhedrosis virus inclusion bodies

    SciTech Connect

    Di, X.; Sun, Y.K.; McCrae, M.A.; Rossmann, M.G. )

    1991-01-01

    Cytoplasmic polyhedrosis virus is an insect reovirus which is occluded in crystalline inclusion bodies that form in the mid-gut of certain insects. Inclusion bodies of cytoplasmic polyhedrosis virus from Bombyx mori were isolated and purified. These crystalline bodies, about 1-3 microns in linear size, were compacted in a capillary tube while immersed in buffer. X-ray diffraction photographs showed powder rings, extending to 8.2 A resolution, which could be indexed with a cell measuring a = b = 49.9 +/- 0.4 A, c = 41.5 +/- 0.4 A, alpha = beta = gamma = 90 degrees. The polyhedrin protein, which is the major component of the inclusion body, has a molecular weight of about 30,000 daltons and, hence, there are probably two molecules in the unit cell. Thus, the unit cell is monoclinic or possibly triclinic. A Patterson derived from the measured powder pattern intensities, assuming monoclinic symmetry, could be interpreted in terms of a molecule with two larger globes. Such a structure is roughly consistent with the breakdown of the polyhedrin into two larger fragments of molecular weight 17,000 and 14,000 when raising the pH to near 10. Under these conditions the inclusion bodies disintegrate, releasing virus and catalyzing the proteolysis of the polyhedrin.

  20. Chemical treatment of Escherichia coli. II. Direct extraction of recombinant protein from cytoplasmic inclusion bodies in intact cells.

    PubMed

    Falconer, R J; O'Neill, B K; Middelberg, A P

    1998-02-20

    A method is presented for the direct extraction of the recombinant protein Long-R3-IGF-I from inclusion bodies located in the cytoplasm of intact Escherichia coli cells. Chemical treatment with 6M urea, 3 mM EDTA, and 20 mM dithiothreitol (DTT) at pH 9.0 proved an effective combination for extracting recombinant protein from intact cells. Comparable levels of Long-R3-IGF-I were recovered by direct extraction as achieved by in vitro dissolution following mechanical disruption. However, the purity of directly extracted recombinant protein was lower due to contamination by bacterial cell components. The kinetics of direct extraction are described using a first-order equation with the time constant of 3 min. Urea appears important for permeabilization of the cell and dissolution of the inclusion body. Conversely, EDTA is involved in permeabilization of the cell wall and DTT enhances protein release. pH proved to be important with lower levels of protein release achieved at low pH values (<9). Cell concentration also had a minor effect on Long-R3-IGF-I release and caused an observable increase in viscosity. Advantages of the direct extraction method include its speed, simplicity, and efficiency at releasing product. PMID:10099214

  1. [Inclusion Body Disease (IBD of Boids)--a haematological, histological and electron microscopical study].

    PubMed

    Keilwerth, Melanie; Bühler, Ilina; Hoffmann, Rudolf; Soliman, Hatem; El-Matbouli, Mansour

    2012-01-01

    Our objective was to evaluate diagnostic tools for the detection of Inclusion Body Disease (IBD) in bold snakes. The aetiology of IBD is unknown, and the disease has non-specific clinical signs, hence there is a need for a clinically-applicable, specific diagnostic method. We examined blood smears and liver biopsies from 26 bold snakes (17 boas and nine pythons; some of which were suspected of having IBD) for the presence of characteristic inclusion bodies. We used haematology, histology and electron microscopy to characterise samples as IBD-positive or -negative. Our results indicate that examination of a simple blood smear is sufficient to diagnose IBD in boas. Inclusion bodies in lymphocytes, erythrocytes and thrombocytes were observed. In both, boas and pythons, we detected inclusion bodies within hepatocytes. We demonstrated also that IBD was more common in boas than in pythons: only samples from two Ball Pythons (Python regius) tested positive, whereas no other Pythonidae were positive. We consider that blood smears represents a rapid, non-invasive technique for detection of IBD. PMID:23045804

  2. Expression of nattokinase in Escherichia coli and renaturation of its inclusion body.

    PubMed

    Ni, He; Guo, Peng-Cheng; Jiang, Wei-Ling; Fan, Xiao-Min; Luo, Xiang-Yu; Li, Hai-Hang

    2016-08-10

    Nattokinase is an important fibrinolytic enzyme with therapeutic applications for cardiovascular diseases. The full-length and mature nattokinase genes were cloned from Bacillus subtilis var. natto and expressed in pQE30 vector in Escherichia coli. The full-length gene expressed low nattokinase activity in the intracellular soluble and the medium fractions. The mature gene expressed low soluble nattokinase activity and large amount insoluble protein in inclusion bodies without enzyme activity. Large amount of refolding solutions (RSs) at different pH values were screening and RS-10 and RS-11 at pH 9 were selected to refold nattokinase inclusion bodies. The recombinant cells were lysed with 0.1mg/mL lysozyme and ultrasonic treatment. After centrifugation, the pellete was washed twice with 20mM Tris-HCl buffer (pH 7.5) containing 1% Triton X-100 to purify the inclusion bodies. The inclusion bodies were dissolved in water at pH 12.0 and refolded with RS-10. The refolded proteins showed 42.8IU/mg and 79.3IU/mg fibrinolytic activity by the traditional dilution method (20-fold dilution into RS-10) and the directly mixing the protein solution with equal volume RS-10, respectively, compared to the 52.0IU/mg of total water-soluble proteins from B. subtilis var. natto. This work demonstrated that the inclusion body of recombinant nattokinase expressed in E. coli could be simply refolded to the natural enzyme activity level by directly mixing the protein solution with equal volume refolding solution. PMID:27234878

  3. Intracytoplasmic inclusion bodies in the chondrocytes of type I lethal achondrogenesis.

    PubMed

    Yang, S S; Heidelberger, K P; Bernstein, J

    1976-11-01

    Lethal achondrogenesis in the past has been frequently confused with achondroplasia. Clinical and radiologic advances in the last decade have led to clear differentiation of this condition from other types of bone dysplasia. It is further separated into two types on the basis of radiographic and pathologic findings. Re-evaluation of the histologic features has led to the recognition of heretofore unrecognized intracytoplasmic inclusion bodies in the chondrocytes of type 1 achondrogenesis. The finding of inclusions strengthens the differentiation of the two types and may have implications for the pathogenesis of the form of chondrodystrophy. PMID:186383

  4. [TDP-43 proteinopathies - from frontotemporal lobar degeneration to inclusion body myositis].

    PubMed

    Kierdaszuk, Biruta; Berdyński, Mariusz; Zekanowski, Cezary; Kamińska, Anna

    2012-01-01

    TDP-43, a newly described neurodegenerative protein, is of great interest to both neurologists and geneticists. At the beginning, its dysfunction was recognized in sporadic amyotrophic lateral sclerosis, frontotemporal lobar degeneration with ubiquitinated inclusions and in mixed forms. However, it was also proved that TDP-43 inclusions are in addition present in many other diseases, for example in inclusion body myositis. Furthermore, many genes and different loci may be involved in pathological TDP-43 accumulation in cells and tissues. Mutations in the TARDPB gene, progranulin gene (PGRNVCP) as well as a gene on chromosome 9p were found. The present paper is a summary on possible involvement of TDP-43 in various neurodegenerative disorders. PMID:23023438

  5. Determining astrophysical three-body radiative capture reaction rates from inclusive Coulomb break-up measurements

    NASA Astrophysics Data System (ADS)

    Casal, J.; Rodríguez-Gallardo, M.; Arias, J. M.; Gómez-Camacho, J.

    2016-04-01

    A relationship between the Coulomb inclusive break-up probability and the radiative capture reaction rate for weakly bound three-body systems is established. This direct link provides a robust procedure to estimate the reaction rate for nuclei of astrophysical interest by measuring inclusive break-up processes at different energies and angles. This might be an advantageous alternative to the determination of reaction rates from the measurement of B (E 1 ) distributions through exclusive Coulomb break-up experiments. In addition, it provides a reference to assess the validity of different theoretical approaches that have been used to calculate reaction rates. The procedure is applied to 11Li (9Li+n +n ) and 6He (4He+n +n ) three-body systems for which some data exist.

  6. Standardization of stain used for diagnosing erythrocytic inclusion body syndrome (EIBS)

    USGS Publications Warehouse

    1987-01-01

    Erythrocytic inclusion body syndrome (EIBS), a viral erythrocytic necrosis (VEN)-like disease, has been observed in several areas in the Northwest. This virus disease is clinically diagnosed by microscopic examination of blood smears for intracytoplasmic erythrocytic inclusion bodies. Fish biologists involved in EIBS diagnostic work have been using several types of hematological stains. It became apparent that standardization of the staining procedure was needed. Comparative tests were conducted on blood smears and kidney imprints with the following commonly used blood stains: (1) Leishman-Giesma, (2) Pinacyanol chloride, (3) Powell 's Giemsa, (4) Harleco's Giemsa, (5) Diff Quik differential stain, (6) Wright's.Pinacyanol chloride stain was found to be the most consistent. The following staining procedure is recommended.

  7. Arenavirus Coinfections Are Common in Snakes with Boid Inclusion Body Disease.

    PubMed

    Hepojoki, J; Salmenperä, P; Sironen, T; Hetzel, U; Korzyukov, Y; Kipar, A; Vapalahti, O

    2015-08-01

    Recently, novel arenaviruses were found in snakes with boid inclusion body disease (BIBD); these form the new genus Reptarenavirus within the family Arenaviridae. We used next-generation sequencing and de novo sequence assembly to investigate reptarenavirus isolates from our previous study. Four of the six isolates and all of the samples from snakes with BIBD contained at least two reptarenavirus species. The viruses sequenced comprise four novel reptarenavirus species and a representative of a new arenavirus genus. PMID:26041290

  8. Arenavirus Coinfections Are Common in Snakes with Boid Inclusion Body Disease

    PubMed Central

    Salmenperä, P.; Sironen, T.; Hetzel, U.; Korzyukov, Y.; Kipar, A.; Vapalahti, O.

    2015-01-01

    Recently, novel arenaviruses were found in snakes with boid inclusion body disease (BIBD); these form the new genus Reptarenavirus within the family Arenaviridae. We used next-generation sequencing and de novo sequence assembly to investigate reptarenavirus isolates from our previous study. Four of the six isolates and all of the samples from snakes with BIBD contained at least two reptarenavirus species. The viruses sequenced comprise four novel reptarenavirus species and a representative of a new arenavirus genus. PMID:26041290

  9. Singular path-independent energy integrals for elastic bodies with thin elastic inclusions

    NASA Astrophysics Data System (ADS)

    Shcherbakov, V. V.

    2016-06-01

    An equilibrium problem for a two-dimensional homogeneous linear elastic body containing a thin elastic inclusion and an interfacial crack is considered. The thin inclusion is modeled within the framework of Euler-Bernoulli beam theory. An explicit formula for the first derivative of the energy functional with respect to the crack perturbation along the interface is presented. It is shown that the formulas for the derivative associated with translation and self-similar expansion of the crack are represented as path-independent integrals along smooth contour surrounding one or both crack tips. These path-independent integrals consist of regular and singular terms and are analogs of the well-known Eshelby-Cherepanov-Rice J-integral and Knowles-Sternberg M-integral.

  10. Actin Recruitment to the Chlamydia Inclusion Is Spatiotemporally Regulated by a Mechanism That Requires Host and Bacterial Factors

    PubMed Central

    Chin, Elizabeth; Kirker, Kelly; Zuck, Meghan; James, Garth; Hybiske, Kevin

    2012-01-01

    The ability to exit host cells at the end of their developmental growth is a critical step for the intracellular bacterium Chlamydia. One exit strategy, extrusion, is mediated by host signaling pathways involved with actin polymerization. Here, we show that actin is recruited to the chlamydial inclusion as a late event, occurring after 20 hours post-infection (hpi) and only within a subpopulation of cells. This event increases significantly in prevalence and extent from 20 to 68 hpi, and actin coats strongly correlated with extrusions. In contrast to what has been reported for other intracellular pathogens, actin nucleation on Chlamydia inclusions did not ‘flash’, but rather exhibited moderate depolymerization dynamics. By using small molecule agents to selectively disrupt host signaling pathways involved with actin nucleation, modulate actin polymerization dynamics and also to disable the synthesis and secretion of chlamydial proteins, we further show that host and bacterial proteins are required for actin coat formation. Transient disruption of either host or bacterial signaling pathways resulted in rapid loss of coats in all infected cells and a reduction in extrusion formation. Inhibition of Chlamydia type III secretion also resulted in rapid loss of actin association on inclusions, thus implicating chlamydial effector proteins(s) as being central factors for engaging with host actin nucleating factors, such as formins. In conclusion, our data illuminate the host and bacterial driven process by which a dense actin matrix is dynamically nucleated and maintained on the Chlamydia inclusion. This late stage event is not ubiquitous for all infected cells in a population, and escalates in prevalence and extent throughout the developmental cycle of Chlamydia, culminating with their exit from the host cell by extrusion. The initiation of actin recruitment by Chlamydia appears to be novel, and may serve as an upstream determinant of the extrusion mechanism. PMID

  11. Radiation-Dependent Limit for the Viability of Bacterial Spores in Halite Fluid Inclusions and on Mars

    PubMed Central

    Kminek, Gerhard; Bada, Jeffrey L.; Pogliano, Kit; Ward, John F.

    2014-01-01

    Kminek, G., Bada, J. L., Pogliano, K. and Ward, J. F. Radiation-Dependent Limit for the Viability of Bacterial Spores in Halite Fluid Inclusions and on Mars. Radiat. Res. 159, 722–729 (2003). When claims for the long-term survival of viable organisms are made, either within terrestrial minerals or on Mars, considerations should be made of the limitations imposed by the naturally occurring radiation dose to which they have been exposed. We investigated the effect of ionizing radiation on different bacterial spores by measuring the inactivation constants for B. subtilis and S. marismortui spores in solution as well as for dry spores of B. subtilis and B. thuringiensis. S. marismortui is a halophilic spore that is genetically similar to the recently discovered 2-9-3 bacterium from a halite fluid inclusion, claimed to be 250 million years old (Vreeland et al., Nature 407, 897–900, 2000). B. thuringiensis is a soil bacterium that is genetically similar to the human pathogens B. anthracis and B. cereus (Helgason et al., Appl. Environ. Microbiol. 66, 2627–2630, 2000). To relate the inactivation constant to some realistic environments, we calculated the radiation regimen in a halite fluid inclusion and in the Martian subsurface over time. Our conclusion is that the ionizing dose of radiation in those environments limits the survival of viable bacterial spores over long periods. In the absence of an active repair mechanism in the dormant state, the long-term survival of spores is limited to less than 109 million years in halite fluid inclusions, to 100 to 160 million years in the Martian subsurface below 3 m, and to less than 600,000 years in the uppermost meter of Mars. PMID:12751954

  12. Isolation, Identification, and Characterization of Novel Arenaviruses, the Etiological Agents of Boid Inclusion Body Disease

    PubMed Central

    Hetzel, Udo; Sironen, Tarja; Laurinmäki, Pasi; Liljeroos, Lassi; Patjas, Aino; Henttonen, Heikki; Vaheri, Antti; Artelt, Annette; Kipar, Anja; Butcher, Sarah J.; Vapalahti, Olli

    2013-01-01

    Boid inclusion body disease (BIBD) is a progressive, usually fatal disease of constrictor snakes, characterized by cytoplasmic inclusion bodies (IB) in a wide range of cell types. To identify the causative agent of the disease, we established cell cultures from BIBD-positive and -negative boa constrictors. The IB phenotype was maintained in cultured cells of affected animals, and supernatants from these cultures caused the phenotype in cultures originating from BIBD-negative snakes. Viruses were purified from the supernatants by ultracentrifugation and subsequently identified as arenaviruses. Purified virus also induced the IB phenotype in naive cells, which fulfilled Koch's postulates in vitro. One isolate, tentatively designated University of Helsinki virus (UHV), was studied in depth. Sequencing confirmed that UHV is a novel arenavirus species that is distinct from other known arenaviruses including those recently identified in snakes with BIBD. The morphology of UHV was established by cryoelectron tomography and subtomographic averaging, revealing the trimeric arenavirus spike structure at 3.2-nm resolution. Immunofluorescence, immunohistochemistry, and immunoblotting with a polyclonal rabbit antiserum against UHV and reverse transcription-PCR (RT-PCR) revealed the presence of genetically diverse arenaviruses in a large cohort of snakes with BIBD, confirming the causative role of arenaviruses. Some snakes were also found to carry arenavirus antibodies. Furthermore, mammalian cells (Vero E6) were productively infected with UHV, demonstrating the potential of arenaviruses to cross species barriers. In conclusion, we propose the newly identified lineage of arenaviruses associated with BIBD as a novel taxonomic entity, boid inclusion body disease-associated arenaviruses (BIBDAV), in the family Arenaviridae. PMID:23926354

  13. Isolation, identification, and characterization of novel arenaviruses, the etiological agents of boid inclusion body disease.

    PubMed

    Hetzel, Udo; Sironen, Tarja; Laurinmäki, Pasi; Liljeroos, Lassi; Patjas, Aino; Henttonen, Heikki; Vaheri, Antti; Artelt, Annette; Kipar, Anja; Butcher, Sarah J; Vapalahti, Olli; Hepojoki, Jussi

    2013-10-01

    Boid inclusion body disease (BIBD) is a progressive, usually fatal disease of constrictor snakes, characterized by cytoplasmic inclusion bodies (IB) in a wide range of cell types. To identify the causative agent of the disease, we established cell cultures from BIBD-positive and -negative boa constrictors. The IB phenotype was maintained in cultured cells of affected animals, and supernatants from these cultures caused the phenotype in cultures originating from BIBD-negative snakes. Viruses were purified from the supernatants by ultracentrifugation and subsequently identified as arenaviruses. Purified virus also induced the IB phenotype in naive cells, which fulfilled Koch's postulates in vitro. One isolate, tentatively designated University of Helsinki virus (UHV), was studied in depth. Sequencing confirmed that UHV is a novel arenavirus species that is distinct from other known arenaviruses including those recently identified in snakes with BIBD. The morphology of UHV was established by cryoelectron tomography and subtomographic averaging, revealing the trimeric arenavirus spike structure at 3.2-nm resolution. Immunofluorescence, immunohistochemistry, and immunoblotting with a polyclonal rabbit antiserum against UHV and reverse transcription-PCR (RT-PCR) revealed the presence of genetically diverse arenaviruses in a large cohort of snakes with BIBD, confirming the causative role of arenaviruses. Some snakes were also found to carry arenavirus antibodies. Furthermore, mammalian cells (Vero E6) were productively infected with UHV, demonstrating the potential of arenaviruses to cross species barriers. In conclusion, we propose the newly identified lineage of arenaviruses associated with BIBD as a novel taxonomic entity, boid inclusion body disease-associated arenaviruses (BIBDAV), in the family Arenaviridae. PMID:23926354

  14. Processing of refractory meteorite inclusions (CAIs) in parent-body atmospheres

    NASA Technical Reports Server (NTRS)

    Podolak, Morris; Bunch, T. E.; Cassen, Pat; Reynolds, Ray T.; Chang, S.

    1990-01-01

    Ca-Al-rich inclusions (CAIs) in refractory meteorites are shown to have been subject to partial melting during a suitably high gas density/small scale height regime arising during gasdynamic deceleration in a temporary atmosphere around an accreting parent body. The presence of dust in such an atmosphere would have increased the pressure gradient with height, lowering the boiloff rate, and permitting dust particles to become trapped in the partially melted material. CAIs may therefore be studied as probes of a primitive atmosphere.

  15. Radiation-dependent limit for the viability of bacterial spores in halite fluid inclusions and on Mars.

    PubMed

    Kminek, Gerhard; Bada, Jeffrey L; Pogliano, Kit; Ward, John F

    2003-06-01

    When claims for the long-term survival of viable organisms are made, either within terrestrial minerals or on Mars, considerations should be made of the limitations imposed by the naturally occurring radiation dose to which they have been exposed. We investigated the effect of ionizing radiation on different bacterial spores by measuring the inactivation constants for B. subtilis and S. marismortui spores in solution as well as for dry spores of B. subtilis and B. thuringiensis. S. marismortui is a halophilic spore that is genetically similar to the recently discovered 2-9-3 bacterium from a halite fluid inclusion, claimed to be 250 million years old (Vreeland et al., Nature 407, 897-900, 2000). B. thuringiensis is a soil bacterium that is genetically similar to the human pathogens B. anthracis and B. cereus (Helgason et al., Appl. Environ. Microbiol. 66, 2627-2630, 2000). To relate the inactivation constant to some realistic environments, we calculated the radiation regimen in a halite fluid inclusion and in the Martian subsurface over time. Our conclusion is that the ionizing dose of radiation in those environments limits the survival of viable bacterial spores over long periods. In the absence of an active repair mechanism in the dormant state, the long-term survival of spores is limited to less than 109 million years in halite fluid inclusions, to 100 to 160 million years in the Martian subsurface below 3 m, and to less than 600,000 years in the uppermost meter of Mars. PMID:12751954

  16. Radiation-Dependent Limit for the Viability of Bacterial Spores in Halite Fluid Inclusions and on Mars

    NASA Technical Reports Server (NTRS)

    Kminek, Gerhard; Bada, Jeffrey L.; Pogliano, Kit; Ward, John F.

    2003-01-01

    When claims for the long-term survival of viable organisms are made, either within terrestrial minerals or on Mars, considerations should be made of the limitations imposed by the naturally occurring radiation dose to which they have been exposed. We investigated the effect of ionizing radiation on different bacterial spores by measuring the inactivation constants for B. subtilis and s. marismortui spores in solution as well as for dry spores of B. subtilis and B. thuringiensis. S. marismortui is a halophilic spore that is genetically similar to the recently discovered 2-9-3 bacterium from a halite fluid inclusion, claimed to be 250 million years old, B. thuringiensis is a soil bacterium that is genetically similar to the human pathogens B. anthracis and B. cereus. To relate the inactivation constant to some realistic environments, we calculated the radiation regimen in a halite fluid inclusion and in the Martian subsurface over time. Our conclusion is that the ionizing dose of radiation in those environments limits the survival of viable bacterial spores over long periods. In the absence of an active repair mechanism in the dormant state, the long-term survival of spores is limited to less than 109 million years in halite fluid inclusions, to 100 to 160 million years in the Martian subsurface below 3 m, and to less than 600,000 years in the upper-most meter of Mars.

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

    PubMed

    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. PMID:21061547

  18. Sporadic inclusion body myositis in Japanese is associated with the MHC ancestral haplotype 52.1.

    PubMed

    Scott, Adrian Phillip; Allcock, Richard James Nigel; Mastaglia, Frank; Nishino, Ichizo; Nonaka, Ikuya; Laing, Nigel

    2006-05-01

    In Caucasians, sporadic inclusion body myositis has been associated with the MHC ancestral haplotypes; HLA-A1, B8, DR3 (8.1AH) and HLA-B35, DR1 (35.2AH). It is not known whether these haplotypes carry susceptibility for the disease in other ethnic groups. We report here the results of HLA-B and -DRB1 typing using a high-resolution sequence-based technique in a cohort of 31 Japanese patients with definite sIBM. Patient allele frequencies were 40.3% for HLA-B*5201 (10.7% in controls: p<0.001) and 37.1% for HLA-DRB1*1502 (10% in controls: p<0.001). Both alleles were found together as part of a conserved haplotype (52.1AH) at a frequency of 37.1% in patients (8.4% in controls: p<0.001). This is the first description of a haplotypic MHC association with sporadic inclusion body myositis in Japanese patients. These findings indicate that different MHC ancestral haplotypes are associated with sIBM in different ethnic groups and further emphasize the importance of genetic factors in this condition. PMID:16564169

  19. Mechanics of swimming of multi-body bacterial swarmers using non-labeled cell tracking algorithm

    NASA Astrophysics Data System (ADS)

    Phuyal, Kiran; Kim, Min Jun

    2013-01-01

    To better understand the survival strategy of bacterial swarmers and the mechanical advantages offered by the linear chain (head-tail) attachment of the multiple bacterial bodies in an individual swarmer cell at low Reynolds number, a non-labeled cell tracking algorithm was used to quantify the mechanics of multi-body flagellated bacteria, Serratia marcescens, swimming in a motility buffer that originally exhibited the swarming motility. Swarming is a type of bacterial motility that is characterized by the collective coordinated motion of differentiated swarmer cells on a two-dimensional surface such as agar. In this study, the bacterial swarmers with multiple cell bodies (2, 3, and 4) were extracted from the swarm plate, and then tracked individually after resuspending in the motility medium. Their motion was investigated and compared with individual undifferentiated swimming bacterial cells. The swarmers when released into the motility buffer swam actively without tumbles. Their speeds, orientations, and the diffusive properties were studied by tracking the individual cell trajectories over a short distance in two-dimensional field when the cells are swimming at a constant depth in a bulk aqueous environment. At short time scales, the ballistic trajectory was dominant for both multi-body swarmers and undifferentiated cells.

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

    PubMed

    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. PMID:26134566

  1. Detection of arenavirus in a peripheral odontogenic fibromyxoma in a red tail boa (Boa constrictor constrictor) with inclusion body disease.

    PubMed

    Hellebuyck, Tom; Pasmans, Frank; Ducatelle, Richard; Saey, Veronique; Martel, An

    2015-03-01

    A captive bred red tail boa (Boa constrictor constrictor) was presented with a large intraoral mass originating from the buccal gingiva, attached to the right dentary teeth row. Based on the clinical features and histological examination, the diagnosis of a peripheral odontogenic fibromyxoma was made. Sections of liver biopsies and circulating lymphocytes contained relatively few eosinophilic intracytoplasmic inclusion bodies, indistinguishable from those observed in inclusion body disease-affected snakes. Inclusion bodies were not observed in cells comprising the neoplastic mass. Using reverse transcription polymerase chain reaction (RT-PCR), arenavirus was detected in the neoplastic tissue. Two years after surgical removal of the mass, recurrence of the neoplastic lesion was observed. Numerous large inclusion body disease inclusions were abundantly present in the neoplastic cells of the recurrent fibromyxoma. Sections of liver biopsies and circulating lymphocytes contained relatively few intracytoplasmic inclusions. The RT-PCR revealed the presence of arenavirus in blood, a liver biopsy, and neoplastic tissue. The present case describes the co-occurrence of an arenavirus infection and an odontogenic fibromyxoma in a red tail boa. PMID:25776548

  2. Efficient production of a correctly folded mouse α-defensin, cryptdin-4, by refolding during inclusion body solubilization.

    PubMed

    Tomisawa, Satoshi; Sato, Yuji; Kamiya, Masakatsu; Kumaki, Yasuhiro; Kikukawa, Takashi; Kawano, Keiichi; Demura, Makoto; Nakamura, Kiminori; Ayabe, Tokiyoshi; Aizawa, Tomoyasu

    2015-08-01

    Mammalian α-defensins contribute to innate immunity by exerting antimicrobial activity against various pathogens. To perform structural and functional analysis of α-defensins, large amounts of α-defensins are essential. Although many expression systems for the production of recombinant α-defensins have been developed, attempts to obtain large amounts of α-defensins have been only moderately successful. Therefore, in this study, we applied a previously developed aggregation-prone protein coexpression method for the production of mouse α-defensin cryptdin-4 (Crp4) in order to enhance the formation of inclusion bodies in Escherichia coli expression system. By using this method, we succeeded in obtaining a large amount of Crp4 in the form of inclusion bodies. Moreover, we attempted to refold Crp4 directly during the inclusion-body solubilization step under oxidative conditions. Surprisingly, even without any purification, Crp4 was efficiently refolded during the solubilization step of inclusion bodies, and the yield was better than that of the conventional refolding method. NMR spectra of purified Crp4 suggested that it was folded into its correct tertiary structure. Therefore, the method described in this study not only enhances the expression of α-defensin as inclusion bodies, but also eliminates the cumbersome and time-consuming refolding step. PMID:25913370

  3. Online analysis of protein inclusion bodies produced in E. coli by monitoring alterations in scattered and reflected light.

    PubMed

    Ude, Christian; Ben-Dov, Nadav; Jochums, André; Li, Zhaopeng; Segal, Ester; Scheper, Thomas; Beutel, Sascha

    2016-05-01

    The online monitoring of recombinant protein aggregate inclusion bodies during microbial cultivation is an immense challenge. Measurement of scattered and reflected light offers a versatile and non-invasive measurement technique. Therefore, we investigated two methods to detect the formation of inclusion bodies and monitor their production: (1) online 180° scattered light measurement (λ = 625 nm) using a sensor platform during cultivation in shake flask and (2) online measurement of the light reflective interference using a porous Si-based optical biosensor (SiPA). It could be shown that 180° scattered light measurement allows monitoring of alterations in the optical properties of Escherichia coli BL21 cells, associated with the formation of inclusion bodies during cultivation. A reproducible linear correlation between the inclusion body concentration of the non-fluorescent protein human leukemia inhibitory factor (hLIF) carrying a thioredoxin tag and the shift ("Δamp") in scattered light signal intensity was observed. This was also observed for the glutathione-S-transferase-tagged green fluorescent protein (GFP-GST). Continuous online monitoring of reflective interference spectra reveals a significant increase in the bacterium refractive index during hLIF production in comparison to a non-induced reference that coincide with the formation of inclusion bodies. These online monitoring techniques could be applied for fast and cost-effective screening of different protein expression systems. PMID:26940052

  4. Hypokalemia-Induced Rhabdomyolysis by Primary Aldosteronism Coexistent With Sporadic Inclusion Body Myositis

    PubMed Central

    Lee, Jong Ha; Chon, Suk

    2015-01-01

    We describes a patient with hypokalemia-induced rhabdomyolysis due to primary aldosteronism (PA), who suffered from slowly progressive muscle weakness after laparoscopic adrenalectomy, and was later diagnosed with coexisting sporadic inclusion body myositis (sIBM). A 54-year-old Asian male presented with severe muscle weakness of both lower extremities. Laboratory findings showed profound hypokalemia, and extreme elevation of the serum creatine phosphokinase levels, suggestive of hypokalemia-induced rhabdomyolysis. Further evaluation strongly suggested PA by an aldosterone-producing adenoma, which was successfully removed surgically. However, muscle weakness slowly progressed one year after the operation and a muscle biopsy demonstrated findings consistent with sIBM. This case is the first report of hypokalemia-induced rhabdomyolysis by PA coexistent with sIBM, to the best of our knowledge. PMID:26605182

  5. KNEE EXTENSOR STRENGTH EXHIBITS POTENTIAL TO PREDICT FUNCTION IN SPORADIC INCLUSION-BODY MYOSITIS

    PubMed Central

    LOWES, LINDA PAX; ALFANO, LINDSAY; VIOLLET, LAURENCE; ROSALES, XIOMARA QUINTERO; SAHENK, ZARIFE; KASPAR, BRIAN K.; CLARK, K. REED; FLANIGAN, KEVIN M.; MENDELL, JERRY R.

    2013-01-01

    Introduction In this study we address the challenging issue of potential use of muscle strength to predict function in clinical trials. This has immediate relevance to translational studies that attempt to improve quadriceps strength in sporadic inclusion-body myositis (sIBM). Methods Maximum voluntary isometric contraction testing as a measure of muscle strength and a battery of functional outcomes were tested in 85 ambulatory subjects with sIBM. Results Marked quadriceps weakness was noted in all patients. Strength was correlated with distance walked at 2 and 6 minutes. Additional correlations were found with time to get up from a chair, climb stairs, and step up on curbs. Conclusions Quadriceps (knee extensor) strength correlated with performance in this large cohort of sIBM subjects, which demonstrated its potential to predict function in this disease. These data provide initial support for use of muscle strength as a surrogate for function, although validation in a clinical trial is required. PMID:22246869

  6. Research note: the isolation of a herpes virus from captive cranes with an inclusion body disease

    USGS Publications Warehouse

    Docherty, D.E.; Henning, D.J.

    1980-01-01

    A viral agent, identified as a herpesvirus and tentatively called 'inclusion body disease of cranes' (IBDC), was isolated from captive cranes involved in a die-off at the International Crane Foundation near Baraboo, Wisconsin. Preliminary animal susceptibility tests, based on experimental infections, suggested that White Pekin ducklings up to 17 days old and adult coots were susceptible to the IBDC virus whereas 16-day-old White Leghorn chicks and 64-day-old Muscovy ducks were not. No serum antibody to IBDC virus was detected in 95 wild sandhill cranes collected in Wisconsin or Indiana in 1976 and 1977. However, 9 of 11 captive cranes in the affected area at the ICF had antibody to this agent.

  7. One-step extraction of functional recombinant aquaporin Z from inclusion bodies with optimal detergent.

    PubMed

    Wang, Lili; Zhou, Hu; Li, Zhengjun; Lim, Teck Kwang; Lim, Xin Shan; Lin, Qingsong

    2015-11-01

    Aquaporins are integral membrane channel proteins found in all kingdoms of life. The Escherichia coli aquaporin Z (AqpZ) has been shown to solely conduct water at high permeability. Functional AqpZ is generally purified from the membrane fraction. However, the quantity of the purified protein is limited. In this study, a new method is developed to achieve high yield of bioactive AqpZ protein. A mild detergent n-dodecyl-β-D-maltopyranoside (DDM) was used to solubilize the over-expressed insoluble AqpZ from inclusion bodies without a refolding process. The recovered AqpZ protein showed high water permeability comparable with AqpZ obtained from the membrane fraction. In this way, the total yield of bioactive AqpZ has been increased greatly, which will facilitate the structural and functional characterization and future applications of AqpZ. PMID:26278820

  8. Experimental infection of Boa constrictor with an orthoreovirus isolated from a snake with inclusion body disease.

    PubMed

    Darke, Sabina; Marschang, Rachel E; Hetzel, Udo; Reinacher, Manfred

    2014-06-01

    Orthoreoviruses have been associated with disease in reptiles, but have not previously been isolated from snakes with inclusion body disease (IBD). An orthoreovirus was isolated from a Boa constrictor diagnosed with IBD and then used to conduct a transmission study to determine the clinical importance of this virus. For the transmission study, 10 juvenile boas were experimentally infected with the isolated orthoreovirus and compared to 5 sham-infected control animals. Orthoreovirus was reisolated for a period of 18 wk after infection and weight gain was reduced in infected snakes. Histological examination showed a mild hepatitis in three of four virologically positive snakes up to 12 wk after infection. Results indicated that the orthoreovirus was moderately pathogenic, but, no evidence was found to indicate that it was the causal agent of IBD. In the light of the discovery of Arenaviruses in some snakes with IBD, it was proposed that orthoreoviruses may play a role in synergistic infection. PMID:25000715

  9. Inclusion body disease of cranes: comparison of pathologic findings in cranes with acquired vs. experimentally induced disease

    USGS Publications Warehouse

    Schuh, J.C.; Sileo, L.; Siegfried, L.M.; Yuill, Thomas M.

    1986-01-01

    Inclusion body disease of cranes was the cause of death in 17 immature and mature cranes of 5 different species in Wisconsin. A herpesvirus of unknown origin was the apparent cause. An isolate of this herpesvirus was used to experimentally infect 3 species of cranes. Macroscopic and microscopic lesions associated with naturally acquired and experimentally induced disease were essentially identical. Multifocal hepatic and splenic necrosis was found in all cranes evaluated. Necrosis of the gastrointestinal tract, thymus, and bursa of Fabricius also was seen in some of the cranes. Eosinophilic intranuclear inclusion bodies often were commonly associated with hepatic lesions, sometimes with the splenic lesions, and rarely with the thymic or gastrointestinal tract lesions. The lesions of this inclusion body disease were similar to those reported for cranes in Austria from which a crane herpesvirus was isolated.

  10. 40 CFR 180.1149 - Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... polyhedrosis virus of Anagrapha falcifera; exemption from the requirement of a tolerance. 180.1149 Section 180... Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from the... polyhedrosis virus of Anagrapha falcifera is exempted from the requirement of a tolerance in or on all...

  11. 40 CFR 180.1149 - Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... polyhedrosis virus of Anagrapha falcifera; exemption from the requirement of a tolerance. 180.1149 Section 180... Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from the... polyhedrosis virus of Anagrapha falcifera is exempted from the requirement of a tolerance in or on all...

  12. 40 CFR 180.1149 - Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... polyhedrosis virus of Anagrapha falcifera; exemption from the requirement of a tolerance. 180.1149 Section 180... Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from the... polyhedrosis virus of Anagrapha falcifera is exempted from the requirement of a tolerance in or on all...

  13. 40 CFR 180.1149 - Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... polyhedrosis virus of Anagrapha falcifera; exemption from the requirement of a tolerance. 180.1149 Section 180... Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from the... polyhedrosis virus of Anagrapha falcifera is exempted from the requirement of a tolerance in or on all...

  14. 40 CFR 180.1149 - Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... polyhedrosis virus of Anagrapha falcifera; exemption from the requirement of a tolerance. 180.1149 Section 180... Inclusion bodies of the multi-nuclear polyhedrosis virus of Anagrapha falcifera; exemption from the... polyhedrosis virus of Anagrapha falcifera is exempted from the requirement of a tolerance in or on all...

  15. Investigation of splicing changes and post-translational processing of LMNA in sporadic inclusion body myositis.

    PubMed

    Luo, Yue-Bei; Mitrpant, Chalermchai; Johnsen, Russell; Fabian, Vicki; Needham, Merrilee; Fletcher, Sue; Wilton, Steve D; Mastaglia, Frank L

    2013-01-01

    Some features of sporadic inclusion body myositis (s-IBM) suggest that there is acceleration of the normal ageing process in muscle tissue. LMNA encodes the nuclear lamina proteins lamin A/C through alternative splicing, and aberrant splicing of exon 11 leads to the premature ageing disease, Hutchinson-Gilford progeria syndrome. Progerin, the pathogenic isoform expressed in HGPS tissues, has also been detected at low levels in tissues of normal individuals with aging. We therefore investigated the alternative splicing of LMNA gene transcripts, and the post-translational processing of prelamin A, in s-IBM and control muscle samples. Age-related low level expression of the progerin transcript was detected in both s-IBM and control muscles, but was not increased in s-IBM and there was no increase in progerin protein or demonstrable accumulation of intermediate prelamin isoforms in the s-IBM muscles. However, an age-related shift in the balance of splicing towards lamin A-related transcripts, which was present in normal muscles, was not found in s-IBM. Our findings indicate that while there are changes in the patterns of LMNA splicing in s-IBM muscle which are probably secondary to the underlying pathological process, it is unlikely that aberrant splicing of exon 11 or defective post-translational processing of prelamin A are involved in the pathogenesis of the disease. PMID:24040437

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

    PubMed

    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. PMID:23857606

  17. Production, characterization, and application of an organic solvent-tolerant lipase present in active inclusion bodies.

    PubMed

    Li, Suxia; Lin, Kang; Pang, Huaiyu; Wu, Yixin; Xu, Jianhe

    2013-01-01

    An organic solvent-tolerant lipase from Serratia marcescens ECU1010 (rSML) was overproduced in Escherichia coli in an insoluble form. High concentrations of both biomass (50 g cell wet weight/L culture broth) and inclusion bodies (10.5 g/L) were obtained by applying a high-cell-density cultivation procedure. Activity assays indicated that the enzymatic activity of rSML reached 600 U/L. After treatment with isopropyl ether for 12 h, the maximum lipase activity reached 6,000 U/L. Scanning electron microscopy and Fourier transform infrared microspectroscopy revealed the activation mechanism of rSML in the presence of organic solvents. rSML was stable in broad ranges of temperatures and pH values, as well as in a series of organic solvents. Besides, rSML showed the best enantioselectivity for the kinetic resolution of (±)-trans-3-(4-methoxyphenyl)glycidic acid methyl ester. These features render the S. marcescens ECU1010 lipase attractive for biotechnological applications in the field of organic synthesis and pharmaceutical industry. PMID:23269633

  18. Complex mitochondrial DNA rearrangements in individual cells from patients with sporadic inclusion body myositis.

    PubMed

    Rygiel, Karolina A; Tuppen, Helen A; Grady, John P; Vincent, Amy; Blakely, Emma L; Reeve, Amy K; Taylor, Robert W; Picard, Martin; Miller, James; Turnbull, Doug M

    2016-06-20

    Mitochondrial DNA (mtDNA) rearrangements are an important cause of mitochondrial disease and age related mitochondrial dysfunction in tissues including brain and skeletal muscle. It is known that different mtDNA deletions accumulate in single cells, but the detailed nature of these rearrangements is still unknown. To evaluate this we used a complementary set of sensitive assays to explore the mtDNA rearrangements in individual cells from patients with sporadic inclusion body myositis, a late-onset inflammatory myopathy with prominent mitochondrial changes. We identified large-scale mtDNA deletions in individual muscle fibres with 20% of cytochrome c oxidase-deficient myofibres accumulating two or more mtDNA deletions. The majority of deletions removed only the major arc but ∼10% of all deletions extended into the minor arc removing the origin of light strand replication (OL) and a variable number of genes. Some mtDNA molecules contained two deletion sites. Additionally, we found evidence of mitochondrial genome duplications allowing replication and clonal expansion of these complex rearranged molecules. The extended spectrum of mtDNA rearrangements in single cells provides insight into the process of clonal expansion which is fundamental to our understanding of the role of mtDNA mutations in ageing and disease. PMID:27131788

  19. A monoclonal antibody to inclusion body disease of cranes virus enabling specific immunohistochemistry and competitive ELISA

    USGS Publications Warehouse

    Letchworth, G.J.; Fishel, J.R.; Hansen, W.R.

    1997-01-01

    Inclusion body disease of cranes (IBDC) herpesvirus kills some infected cranes and persists in convalescent animals. To enable further study and rapid identification of carrier animals, we developed a monoclonal antibody (MAb) to IBDC virus and used it in immunohistochemistry and a competitive enzyme-linked immunosorbent assay (ELISA). We used conventional techniques to make murine MAbs directed against IBDC virus purified from infected duck embryo cells. Hybridomas reacting in an ELISA with IBDC virus but not uninfected duck embryo cells were characterized by radioimmunoprecipitation, in situ immunohistochemistry, and competitive ELISA with neutralizing and nonneutralizing crane sera. MAb 2C11 immunoprecipitated 59-, 61-, and 110-kD proteins from IBDC virus-infected but not uninfected cells and stained glutaraldehyde-fixed IBDC virus plaques but not surrounding uninfected duck embryo cells in vitro. Antibody 2C11 did not react with duck embryo cells infected with falcon herpesvirus, psittacine herpesvirus, infectious laryngotracheitis, pigeon herpesvirus, or duck plague virus. A competitive ELISA using antibody 2C11 identified most sera that were positive in the neutralization test. This antibody will be useful in further characterizing IBDC virus, its pathogenesis, and its natural history.

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

    PubMed Central

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

    2016-01-01

    Introduction 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. Case Presentation 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. Conclusions 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. PMID:27247916

  1. Controlled Localization of Functionally Active Proteins to Inclusion Bodies Using Leucine Zippers

    PubMed Central

    Choi, Su-Lim; Lee, Sang Jun; Yeom, Soo-Jin; Kim, Hyun Ju; Rhee, Young Ha; Jung, Heung-Chae; Lee, Seung-Goo

    2014-01-01

    Inclusion bodies (IBs) are typically non-functional particles of aggregated proteins. However, some proteins in fusion with amyloid-like peptides, viral coat proteins, and cellulose binding domains (CBDs) generate IB particles retaining the original functions in cells. Here, we attempted to generate CBD IBs displaying functional leucine zipper proteins (LZs) as bait for localizing cytosolic proteins in E. coli. When a red fluorescent protein was tested as a target protein, microscopic observations showed that the IBs red-fluoresced strongly. When different LZ pairs with KDs of 8–1,000 µM were tested as the bait and prey, the localization of the red fluorescence appeared to change following the affinities between the LZs, as observed by fluorescence imaging and flow cytometry. This result proposed that LZ-tagged CBD IBs can be applied as an in vivo matrix to entrap cytosolic proteins in E. coli while maintaining their original activities. In addition, easy detection of localization to IBs provides a unique platform for the engineering and analyses of protein-protein interactions in E. coli. PMID:24897378

  2. Complex mitochondrial DNA rearrangements in individual cells from patients with sporadic inclusion body myositis

    PubMed Central

    Rygiel, Karolina A.; Tuppen, Helen A.; Grady, John P.; Vincent, Amy; Blakely, Emma L.; Reeve, Amy K.; Taylor, Robert W.; Picard, Martin; Miller, James; Turnbull, Doug M.

    2016-01-01

    Mitochondrial DNA (mtDNA) rearrangements are an important cause of mitochondrial disease and age related mitochondrial dysfunction in tissues including brain and skeletal muscle. It is known that different mtDNA deletions accumulate in single cells, but the detailed nature of these rearrangements is still unknown. To evaluate this we used a complementary set of sensitive assays to explore the mtDNA rearrangements in individual cells from patients with sporadic inclusion body myositis, a late-onset inflammatory myopathy with prominent mitochondrial changes. We identified large-scale mtDNA deletions in individual muscle fibres with 20% of cytochrome c oxidase-deficient myofibres accumulating two or more mtDNA deletions. The majority of deletions removed only the major arc but ∼10% of all deletions extended into the minor arc removing the origin of light strand replication (OL) and a variable number of genes. Some mtDNA molecules contained two deletion sites. Additionally, we found evidence of mitochondrial genome duplications allowing replication and clonal expansion of these complex rearranged molecules. The extended spectrum of mtDNA rearrangements in single cells provides insight into the process of clonal expansion which is fundamental to our understanding of the role of mtDNA mutations in ageing and disease. PMID:27131788

  3. Demographic and clinical features of inclusion body myositis in North America

    PubMed Central

    Paltiel, A. David; Ingvarsson, Einar; Lee, Donald K. K.; Leff, Richard L.; Nowak, Richard J.; Petschke, Kurt D.; Richards-Shubik, Seth; Zhou, Ange; Shubik, Martin; O’Connor, Kevin C.

    2016-01-01

    Objective Define the demographics, natural history, and clinical management of patients with inclusion body myositis (IBM). Background Few studies of the demographics, natural history, and clinical management of IBM have been performed in a large patient population. Methods A cross-sectional, self-reporting survey was conducted. Results The mean age of the 916 participants was 70.4 years, the male-to-female ratio was 2:1, and the majority reported difficulty with ambulation and activities of daily living. The earliest symptoms included impaired use and weakness of arms and legs. The mean time from first symptoms to diagnosis was 4.7 years. Half reported that IBM was their initial diagnosis. A composite functional index negatively associated with age, disease duration, and positively associated with participation in exercise. Conclusion These data are valuable for informing patients how IBM manifestations are expected to impair daily living and indicate that self-reporting could be used to establish outcome measures in clinical trials. PMID:25557419

  4. Efficacy of immunosuppressive treatment in a systemic lupus erythematosus patient presenting with inclusion body myositis.

    PubMed

    Varela-Rosario, Noemí; Pérez-Berenguer, Juan L; Vilá, Luis M

    2016-01-01

    Inclusion body myositis (IBM) is an inflammatory myopathy that is generally unresponsive to immunosuppressive drugs. The coexistence of IBM with other autoimmune connective tissue diseases is rare. We present a case of a 76-year-old woman with systemic lupus erythematosus (SLE) who developed proximal muscle weakness of lower extremities and mild elevation of serum creatine kinase (CK) at 495 U/L. Muscle biopsy showed changes of endomysial inflammation and rimmed vacuoles consistent with IBM. She was treated with prednisone 40 mg daily and methotrexate 12.5 mg weekly. One month later, her physical examination showed minimal proximal weakness of lower extremities. CK levels decreased to 44 U/L. Prednisone dose was gradually decreased to 5.0 mg daily. She remained stable with normal CK levels during a follow-up period of 10 months. This case, together with other reports, suggests that IBM in the setting of SLE represents a different subtype that can benefit from immunosuppressive treatment. PMID:27048400

  5. Folding and Purification of Insoluble (Inclusion Body) Proteins from Escherichia coli.

    PubMed

    Wingfield, Paul T; Palmer, Ira; Liang, Shu-Mei

    2014-01-01

    Heterologous expression of recombinant proteins in E. coli often results in the formation of insoluble and inactive protein aggregates, commonly referred to as inclusion bodies. To obtain the native (i.e., correctly folded) and hence active form of the protein from such aggregates, four steps are usually followed: (1) the cells are lysed, (2) the cell wall and outer membrane components are removed, (3) the aggregates are solubilized (or extracted) with strong protein denaturants, and (4) the solubilized, denatured proteins are folded with concomitant oxidation of reduced cysteine residues into the correct disulfide bonds to obtain the native protein. This unit features three different approaches to the final step of protein folding and purification. In the first, guanidine·HCl is used as the denaturant, after which the solubilized protein is folded (before purification) in an "oxido-shuffling" buffer system to increase the rate of protein oxidation. In the second, acetic acid is used to solubilize the protein, which is then partially purified by gel filtration before folding; the protein is then folded and oxidized by simple dialysis against water. Thirdly, folding and purification of a fusion protein using metal-chelate affinity chromatography are described. PMID:25367010

  6. Single pH buffer refolding screen for protein from inclusion bodies.

    PubMed

    Coutard, Bruno; Danchin, Etienne G J; Oubelaid, Rachid; Canard, Bruno; Bignon, Christophe

    2012-04-01

    We previously reported the set up of an automated test for screening the refolding of recombinant proteins expressed as inclusion bodies in Escherichia coli[1]. The screen used 96 refolding buffers and was validated with 24 proteins, 70% of which remained soluble in at least one buffer. In the present paper, we have analyzed in more detail these experimental data to see if the refolding process can be driven by general rules. Notably, we found that proteins with an acidic isoelectric point (pI) refolded in buffers the average pH of which was alkaline and conversely. In addition, the number of refolding buffers wherein a protein remained soluble increased with the difference between its pI and the average pH of the buffers in which it refolded. A trend analysis of the other variables (ionic strength, detergents, etc.) was also performed. On the basis of this analysis, we devised and validated a new refolding screen made of a single buffer for acidic proteins and a single buffer for alkaline proteins. PMID:22343064

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

    PubMed Central

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

    2014-01-01

    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. PMID:24843142

  8. Lymphoblastic lymphoma and leukemic blood profile in a red-tail boa (Boa constrictor constrictor) with concurrent inclusion body disease.

    PubMed

    Schilliger, Lionel; Selleri, Paolo; Frye, Fredric L

    2011-01-01

    An adult male wild-caught true red-tail boa (Boa constrictor constrictor), imported from Surinam, was presented for anorexia, extreme lethargy, and coelomic swelling in the cranial third of the body, in the anatomic location of the thymus. The snake died a few minutes after blood sampling via cardiocentesis. Hematology revealed anemia and extreme leukocytosis (820 × 10(3)/ml) characterized by a predominance (95%) of lymphocytes. Necropsy revealed enlargement of most of the visceral organs. Histology confirmed lymphoblastic lymphoma with a leukemic blood profile and diffuse infiltration of some of the heart, thymus, bone marrow, kidney, spleen, lung, and liver. Several large intracytoplasmic eosinophilic inclusion bodies surrounded by narrow clear "halos" were identified within gastric mucosal cells, proximal and distal convoluted tubule epithelial cells, and splenic cells. The final diagnosis was lymphoblast lymphoma with a leukemic blood profile and concurrent inclusion body disease. PMID:21217051

  9. Twisted tubulofilaments of inclusion body myositis muscle resemble paired helical filaments of Alzheimer brain and contain hyperphosphorylated tau.

    PubMed Central

    Askanas, V.; Engel, W. K.; Bilak, M.; Alvarez, R. B.; Selkoe, D. J.

    1994-01-01

    We immunostained muscle biopsies of 8 patients with sporadic inclusion body myositis (S-IBM), 7 patients with autosomal recessive hereditary inclusion body myopathy (H-IBM) (both diseases being characterized by similar muscle fiber vacuoles containing inclusions), and 11 normal and disease controls. We used the following well-characterized antibodies against tau protein: Tau-1, Alz-50, and anti-paired helical filament (PHF) antiserum. By light microscopy, in all S-IBM muscle biopsies virtually all vacuoles immunoreactive for ubiquitin and beta-amyloid protein also contained inclusions immunoreactive with Alz-50 and anti-PHF antiserum. With tau-1 antibody, strong immunoreactivity in the vacuoles was obtained only after dephosphorylation of muscle sections. By electronmicroscopy, all three antibodies immunodecorated exclusively cytoplasmic twisted tubulofilaments (TTFs). In H-IBM, virtually all ubiquitin and beta-amyloid-positive muscle fiber vacuoles contained inclusions immunoreactive with anti-PHF antiserum, but in only 40% of those fibers were the inclusions immunoreactive with Alz-50. In six H-IBM patients there were no tau-1 immunoreactive inclusions in any of their vacuolated muscle fibers; in one patient, 24% of the vacuolated fibers had tau-1 immunoreactivity. By demonstrating that hyperphosphorylated tau, which is characteristic of Alzheimer brain PHFs, is a component of S-IBM-muscle TTFs (which are also ultrastructurally similar to PHFs), our study: 1) provides the first demonstration of abnormally accumulated tau in nonneural tissue and 2) suggests that the cytopathogenesis in Alzheimer brain and S-IBM muscle may share some similar mechanisms. Whether the difference in tau immunoreactivity between S-IBM and most of the H-IBM patients reflects a difference in genetically determined transcriptional or posttranslational modifications of tau protein or other factors remains to be determined. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:8291607

  10. Nuclear inclusions mimicking poly(A)-binding protein nuclear 1 inclusions in a case of inclusion body myopathy associated with Paget disease of bone and frontotemporal dementia with a novel mutation in the valosin-containing protein gene.

    PubMed

    Matsubara, Shiro; Shimizu, Toshio; Komori, Takashi; Mori-Yoshimura, Madoka; Minami, Narihiro; Hayashi, Yukiko K

    2016-07-01

    A middle-aged Japanese man presented with slowly progressive asymmetric weakness of legs and arm but had neither ptosis nor dysphagia. He had a family history of similar condition suggestive of autosomal dominant inheritance. A muscle biopsy showed mixture of neurogenic atrophy and myopathy with rimmed vacuoles. Furthermore we found intranuclear inclusions that had a fine structure mimicking that of inclusions reported in oculopharyngeal muscular dystrophy (OPMD). Immunohistochemical staining for polyadenylate-binding nuclear protein 1, which is identified within the nuclear inclusions of OPMD, demonstrated nuclear positivity in this case. However, OPMD was thought unlikely based on the clinical features and results of genetic analyses. Instead, a novel mutation in valosin-containing protein, c.376A>T (p.Ile126Phe), was revealed. A diagnosis of inclusion body myopathy associated with Paget disease of bone and frontotemporal dementia was made. This is the first report of polyadenylate-binding nuclear protein 1-positive nuclear inclusions in the muscle of this condition. PMID:27209344

  11. Expression of the inclusion body myopathy 3 mutation in Drosophila depresses myosin function and stability and recapitulates muscle inclusions and weakness.

    PubMed

    Wang, Yang; Melkani, Girish C; Suggs, Jennifer A; Melkani, Anju; Kronert, William A; Cammarato, Anthony; Bernstein, Sanford I

    2012-06-01

    Hereditary myosin myopathies are characterized by variable clinical features. Inclusion body myopathy 3 (IBM-3) is an autosomal dominant disease associated with a missense mutation (E706K) in the myosin heavy chain IIa gene. Adult patients experience progressive muscle weakness. Biopsies reveal dystrophic changes, rimmed vacuoles with cytoplasmic inclusions, and focal disorganization of myofilaments. We constructed a transgene encoding E706K myosin and expressed it in Drosophila (E701K) indirect flight and jump muscles to establish a novel homozygous organism with homogeneous populations of fast IBM-3 myosin and muscle fibers. Flight and jump abilities were severely reduced in homozygotes. ATPase and actin sliding velocity of the mutant myosin were depressed >80% compared with wild-type myosin. Light scattering experiments and electron microscopy revealed that mutant myosin heads bear a dramatic propensity to collapse and aggregate. Thus E706K (E701K) myosin appears far more labile than wild-type myosin. Furthermore, mutant fly fibers exhibit ultrastructural hallmarks seen in patients, including cytoplasmic inclusions containing aberrant proteinaceous structures and disorganized muscle filaments. Our Drosophila model reveals the unambiguous consequences of the IBM-3 lesion on fast muscle myosin and fibers. The abnormalities observed in myosin function and muscle ultrastructure likely contribute to muscle weakness observed in our flies and patients. PMID:22496423

  12. An autopsy case of frontotemporal lobar degeneration with the appearance of fused in sarcoma inclusions (basophilic inclusion body disease) clinically presenting corticobasal syndrome.

    PubMed

    Matsumoto, Arifumi; Suzuki, Hiroyoshi; Fukatsu, Reiko; Shimizu, Hiroshi; Suzuki, Yasushi; Hisanaga, Kinya

    2016-02-01

    We describe an autopsy case of basophilic inclusion body disease (BIBD), a subtype of frontotemporal lobar degeneration (FTLD) with the appearance of fused in sarcoma (FUS) inclusions (FTLD-FUS), clinically presenting corticobasal syndrome (CBS). A 54-year-old man initially developed worsening of stuttering and right hand clumsiness. Neurological examinations revealed rigidity in the right upper and lower extremities, buccofacial apraxia, and right-side dominant limb-kinetic and ideomotor apraxia. Neuroimaging showed asymmetric left-dominant brain atrophy and a cerebral blood flow reduction in the ipsilateral frontal region. At 56 years, his apraxia had advanced, and ideational apraxia was observed. Furthermore, the asymmetry in the limb-kinetic and ideomotor apraxia had disappeared, and both conditions had become bilateral. He had a new onset of aphasia. His symptoms progressed and he died 9 years after the initial symptoms. The brain weighed 955 g. Diffuse brain atrophy was most obvious in the bilateral frontotemporal regions. The atrophy of the left superior frontal and precentral gyri and bilateral basal ganglia was remarkable. Histologically, there was a marked loss of neurons with gliosis in the affected areas, where basophilic neuronal cytoplasmic inclusions were observed. The inclusions were immunoreactive for FUS, p62, and TATA-binding protein-associated factor 15 (TAF15), but not for phosphorylated tau, transactive response DNA-binding protein of 43 kDa (TDP-43), neurofilament protein, or Ewing sarcoma (EWS). From these pathological findings, this case was diagnosed as having BIBD as an FTLD-FUS variant. Spinal cord lower motor neurons were spared in number, similar to primary lateral sclerosis. Mutations in FUS were undetectable. Common background pathologies for CBS include corticobasal degeneration, Alzheimer's disease, PSP, FTLD with phosphorylated TDP-43 inclusions (FTLD-TDP), Pick's disease, Lewy body disease and CJD. However, FTLD-FUS (BIBD

  13. Isolated inclusion body myopathy caused by a multisystem proteinopathy–linked hnRNPA1 mutation

    PubMed Central

    Izumi, Rumiko; Warita, Hitoshi; Niihori, Tetsuya; Takahashi, Toshiaki; Tateyama, Maki; Suzuki, Naoki; Nishiyama, Ayumi; Shirota, Matsuyuki; Funayama, Ryo; Nakayama, Keiko; Mitsuhashi, Satomi; Nishino, Ichizo; Aoki, Yoko

    2015-01-01

    Objective: To identify the genetic cause of isolated inclusion body myopathy (IBM) with autosomal dominant inheritance in 2 families. Methods: Genetic investigations were performed using whole-exome and Sanger sequencing of the heterogeneous nuclear ribonucleoprotein A1 gene (hnRNPA1). The clinical and pathologic features of patients in the 2 families were evaluated with neurologic examinations, muscle imaging, and muscle biopsy. Results: We identified a missense p.D314N mutation in hnRNPA1, which is also known to cause familial amyotrophic lateral sclerosis, in 2 families with IBM. The affected individuals developed muscle weakness in their 40s, which slowly progressed toward a limb-girdle pattern. Further evaluation of the affected individuals revealed no apparent motor neuron dysfunction, cognitive impairment, or bone abnormality. The muscle pathology was compatible with IBM, lacking apparent neurogenic change and inflammation. Multiple immunohistochemical analyses revealed the cytoplasmic aggregation of hnRNPA1 in close association with autophagosomes and myonuclei. Furthermore, the aberrant accumulation was characterized by coaggregation with ubiquitin, sequestome-1/p62, valosin-containing protein/p97, and a variety of RNA-binding proteins (RBPs). Conclusions: The present study expands the clinical phenotype of hnRNPA1-linked multisystem proteinopathy. Mutations in hnRNPA1, and possibly hnRNPA2B1, will be responsible for isolated IBM with a pure muscular phenotype. Although the mechanisms underlying the selective skeletal muscle involvement remain to be elucidated, the immunohistochemical results suggest a broad sequestration of RBPs by the mutated hnRNPA1. PMID:27066560

  14. Cholera toxin B subunit pentamer reassembled from Escherichia coli inclusion bodies for use in vaccination.

    PubMed

    Tamaki, Yukihiro; Harakuni, Tetsuya; Yamaguchi, Rui; Miyata, Takeshi; Arakawa, Takeshi

    2016-03-01

    The cholera toxin B subunit (CTB) is secreted in its pentameric form from Escherichia coli if its leader peptide is replaced with one of E. coli origin. However, the secretion of the pentamer is generally severely impaired when the molecule is mutated or fused to a foreign peptide. Therefore, we attempted to regenerate pentameric CTB from the inclusion bodies (IBs) of E. coli. Stepwise dialysis of the IBs solubilized in guanidine hydrochloride predominantly generated soluble high-molecular-mass (HMM) aggregates and only a small fraction of pentamer. Three methods to reassemble homogeneous pentameric molecules were evaluated: (i) using a pentameric coiled-coil fusion partner, expecting it to function as an assembly core; (ii) optimizing the protein concentration during refolding; and (iii) eliminating contaminants before refolding. Coiled-coil fusion had some effect, but substantial amounts of HMM aggregates were still generated. Varying the protein concentration from 0.05 mg/mL to 5mg/mL had almost no effect. In contrast, eliminating the contaminants before refolding had a robust effect, and only the pentamer was regenerated, with no detectable HMM aggregates. Surprisingly, the protein concentration at refolding was up to 5mg/mL when the contaminants were removed, with no adverse effects on refolding. The regenerated pentamer was indistinguishable in its biochemical and immunological characteristics from CTB secreted from E. coli or choleragenoid from Vibrio cholerae. This study provides a simple but very efficient strategy for pentamerizing CTB with a highly homogeneous molecular conformation, with which it may be feasible to engineer CTB derivatives and CTB fusion antigens. PMID:26828455

  15. Improving the refolding efficiency for proinsulin aspart inclusion body with optimized buffer compositions.

    PubMed

    Chen, Ying; Wang, Qi; Zhang, Chun; Li, Xiunan; Gao, Qiang; Dong, Changqing; Liu, Yongdong; Su, Zhiguo

    2016-06-01

    Successfully recovering proinsulin's native conformation from inclusion body is the crucial step to guarantee high efficiency for insulin's manufacture. Here, two by-products of disulfide-linked oligomers and disulfide-isomerized monomers were clearly identified during proinsulin aspart's refolding through multiple analytic methods. Arginine and urea are both used to assist in proinsulin refolding, however the efficacy and possible mechanism was found to be different. The oligomers formed with urea were of larger size than with arginine. With the urea concentrations increasing from 2 M to 4 M, the content of oligomers decreased greatly, but simultaneously the refolding yield at the protein concentration of 0.5 mg/mL decreased from 40% to 30% due to the increase of disulfide-isomerized monomers. In contrast, with arginine concentrations increasing up to 1 M, the refolding yield gradually increased to 50% although the content for oligomers also decreased. Moreover, it was demonstrated that not redox pairs but only oxidant was necessary to facilitate the native disulfide bonds formation for the reduced denatured proinsulin. An oxidative agent of selenocystamine could increase the yield up to 80% in the presence of 0.5 M arginine. Further study demonstrated that refolding with 2 M urea instead of 0.5 M arginine could achieve similar yield as protein concentration is slightly reduced to 0.3 mg/mL. In this case, refolded proinsulin was directly purified through one-step of anionic exchange chromatography, with a recovery of 32% and purity up to 95%. All the results could be easily adopted in insulin's industrial manufacture for improving the production efficiency. PMID:26826314

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

    PubMed Central

    Sparks, Susan; Rakocevic, Goran; Joe, Galen; Manoli, Irini; Shrader, Joseph; Harris-Love, Michael; Sonies, Barbara; Ciccone, Carla; Dorward, Heidi; Krasnewich, Donna; Huizing, Marjan; Dalakas, Marinos C; Gahl, William A

    2007-01-01

    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. PMID:17261181

  17. Molecular events linking cholesterol to Alzheimer's disease and inclusion body myositis in a rabbit model.

    PubMed

    Liu, Qing Yan; Koukiekolo, Roger; Zhang, Dong Ling; Smith, Brandon; Ly, Dao; Lei, Joy X; Ghribi, Othman

    2016-01-01

    Alzheimer's disease (AD) is the most common neurodegenerative disorder, characterized by cognitive impairment and dementia, resulting from progressive synaptic dysfunction, loss and neuronal cell death. Inclusion body myositis (IBM) is a skeletal muscle degenerative disease, displaying progressive proximal and distal muscle weakness, in association with muscle fiber atrophy, degeneration and death. Studies have shown that the late onset version of AD (LOAD) and sporadic IBM (sIBM) in muscle share many pathological features, including the presence of extracellular plaques of β-amyloid peptides and intracellular tangles of hyperphosphorylated tau proteins. High blood cholesterol is suggested to be a risk factor for LOAD. Many neuropathological changes of LOAD can be reproduced by feeding rabbits a 2% enriched cholesterol diet for 12 weeks. The cholesterol fed rabbit model also simultaneously develops sIBM like pathology, which makes it an ideal model to study the molecular mechanisms common to the development of both diseases. In the present study, we determined the changes of gene expression in rabbit brain and muscle during the progression of LOAD and sIBM pathology using a custom rabbit nucleotide microarray, followed by qRT-PCR analyses. Out of 869 unique transcripts screened, 47 genes showed differential expression between the control and the cholesterol-treated group during the 12 week period and 19 changed transcripts appeared to be common to LOAD and sIBM. The most notable changes are the upregulation of the hemoglobin gene family and the downregulation of the genes required for mitochondrial oxidative phosphorylation in both brain and muscle tissues throughout the time course. The significant overlap on the changes of gene expression in the brain and muscle of rabbits fed with cholesterol-enriched diet supports the notion that LOAD and sIBM may share a common etiology. PMID:27073745

  18. Bacterial Community Variation in Human Body Habitats Across Space and Time

    PubMed Central

    Costello, Elizabeth K.; Lauber, Christian L.; Hamady, Micah; Fierer, Noah; Gordon, Jeffrey I.; Knight, Rob

    2010-01-01

    Elucidating the biogeography of bacterial communities on the human body is critical for establishing healthy baselines from which to detect differences associated with diseases. To obtain an integrated view of the spatial and temporal distribution of the human microbiota, we surveyed bacteria from up to 27 sites in 7–9 healthy adults on four occasions. We found that community composition was determined primarily by body habitat. Within habitats, interpersonal variability was high, while individuals exhibited minimal temporal variability. Several skin locations harbored more diverse communities than the gut and mouth, and skin locations differed in their community assembly patterns. These results indicate that our microbiota, although personalized, varies systematically across body habitats and time: such trends may ultimately reveal how microbiome changes cause or prevent disease. PMID:19892944

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

    SciTech Connect

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

    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

  20. Inclusion bodies: formation and degeneration of the oocytes in the fish Channa punctatus (Bloch) in response to ammonium sulfate treatment

    SciTech Connect

    Ram, R.N.; Sathyanesan, A.G.

    1986-06-01

    In the adult Channa punctatus exposed to 500 ppm of the commonly used fertilizer ammonium sulfate for 6 months from January to June, ovarian growth was retarded significantly. The inhibition of the ovarian growth is reflected on the gonadosomatic index which was significantly reduced as compared with control. Apart from this, in treated fish, several stage-I oocytes exhibited proteinaceous extra- and intranuclear inclusion bodies which are apparently due to cumulative toxic effect of ammonium sulfate. These oocytes ultimately degenerate.

  1. Iron- and 4-hydroxy-2-alkylquinoline-containing periplasmic inclusion bodies of Pseudomonas aeruginosa: A chemical analysis

    USGS Publications Warehouse

    Royt, P.W.; Honeychuck, R.V.; Pant, R.R.; Rogers, M.L.; Asher, L.V.; Lloyd, J.R.; Carlos, W.E.; Belkin, H.E.; Patwardhan, S.

    2007-01-01

    Dark aggregated particles were seen on pellets of iron-rich, mid-logarithmic phase Pseudomonas aeruginosa. Transmission electron microscopy of these cells showed inclusion bodies in periplasmic vacuoles. Aggregated particles isolated from the spent medium of these cells contained iron as indicated by atomic absorption spectroscopy and by electron paramagnetic resonance spectroscopy that revealed Fe3+. Scanning electron microscopy/energy dispersive X-ray analysis of whole cells revealed the presence of iron-containing particles beneath the surface of the cell, indicating that the isolated aggregates were the intracellular inclusion bodies. Collectively, mass spectroscopy and nuclear magnetic resonance spectroscopy of the isolated inclusion bodies revealed the presence of 3,4-dihydroxy-2-heptylquinoline which is the Pseudomonas quinolone signaling compound (PQS) and an iron chelator; 4-hydroxy-2-heptylquinoline (pseudan VII), which is an iron chelator, antibacterial compound and precursor of PQS; 4-hydroxy-2-nonylquinoline (pseudan IX) which is an iron chelator and antibacterial compound; 4-hydroxy-2-methylquinoline (pseudan I), and 4-hydroxy-2-nonylquinoline N-oxide. ?? 2006 Elsevier Inc. All rights reserved.

  2. Inhibition of HDAC6 modifies tau inclusion body formation and impairs autophagic clearance.

    PubMed

    Leyk, Janina; Goldbaum, Olaf; Noack, Monika; Richter-Landsberg, Christiane

    2015-04-01

    Proteinaceous inclusions in nerve cells and glia are a defining neuropathological hallmark in a variety of neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD). Their occurrence may be related to malfunctions of the proteolytic degradation systems. In cultured oligodendrocytes, proteasomal inhibition leads to protein aggregate formation resembling coiled bodies, which are characteristic for PSP and CBD. Large protein aggregates are excluded from the proteasome and can only be degraded by autophagy, a lysosomal pathway. Autophagy is a highly selective process, which requires a variety of receptor proteins for ubiquitinated proteins, such as p62 and histone deacetylase 6 (HDAC6). HDAC6 is mainly localized in the cytoplasm, and alpha-tubulin is its major substrate. HDAC6 is considered as a sensor of proteasomal stress; it is involved in the autophagosomal pathway and can mediate the retrograde transport of ubiquitinated proteins along the microtubules. As we have shown recently, HDAC6 is present in oligodendrocytes and its inhibition leads to morphological alterations, microtubule bundling, modulation of acetylation, and phosphorylation of the microtubule-associated protein tau. The present study was undertaken to investigate whether HDAC6 is involved in protein aggregate formation in oligodendrocytes and whether its inhibition modifies the consequences of MG-132-induced inhibition of the ubiquitin proteasome system (UPS). The data show that HDAC6 and acetylated tau are recruited to protein aggregates after proteasomal inhibition. Pharmacological inhibition of HDAC6 by the selective inhibitor tubastatin A (TST) and its small hairpin RNA (shRNA)-mediated downregulation alters the assembly of MG-132-induced compact protein aggregates. After TST treatment, they appear more diffusely dispersed throughout the cytoplasm. This is not a protective means but promotes the onset

  3. A role for plant microtubules in the formation of transmission-specific inclusion bodies of Cauliflower mosaic virus.

    PubMed

    Martinière, Alexandre; Gargani, Daniel; Uzest, Marilyne; Lautredou, Nicole; Blanc, Stéphane; Drucker, Martin

    2009-04-01

    Interactions between microtubules and viruses play important roles in viral infection. The best-characterized examples involve transport of animal viruses by microtubules to the nucleus or other intracellular destinations. In plant viruses, most work to date has focused on interaction between viral movement proteins and the cytoskeleton, which is thought to be involved in viral cell-to-cell spread. We show here, in Cauliflower mosaic virus (CaMV)-infected plant cells, that viral electron-lucent inclusion bodies (ELIBs), whose only known function is vector transmission, require intact microtubules for their efficient formation. The kinetics of the formation of CaMV-related inclusion bodies in transfected protoplasts showed that ELIBs represent newly emerging structures, appearing at late stages of the intracellular viral life cycle. Viral proteins P2 and P3 are first produced in multiple electron-dense inclusion bodies, and are later specifically exported to transiently co-localize with microtubules, before concentrating in a single, massive ELIB in each infected cell. Treatments with cytoskeleton-affecting drugs suggested that P2 and P3 might be actively transported on microtubules, by as yet unknown motors. In addition to providing information on the intracellular life cycle of CaMV, our results show that specific interactions between host cell and virus may be dedicated to a later role in vector transmission. More generally, they indicate a new unexpected function for plant cell microtubules in the virus life cycle, demonstrating that microtubules act not only on immediate intracellular or intra-host phenomena, but also on processes ultimately controlling inter-host transmission. PMID:19077170

  4. Solubilization of growth hormone and other recombinant proteins from Escherichia coli inclusion bodies by using a cationic surfactant.

    PubMed Central

    Puri, N K; Crivelli, E; Cardamone, M; Fiddes, R; Bertolini, J; Ninham, B; Brandon, M R

    1992-01-01

    Recombinant pig growth hormone (rPGH) was solubilized from inclusion bodies by using the cationic surfactant cetyltrimethylammonium chloride (CTAC). The solubilizing action of CTAC appeared to be dependent on the presence of a positively charged head group, as a non-charged variant was inactive. Relatively low concentrations of CTAC were required for rapid solubilization, and protein-bound CTAC was easily removed by ion-exchange chromatography. Compared with solubilization and recovery of rPGH from inclusion bodies with 7.5 M-urea and 6 M-guanidinium chloride, the relative efficiency of solubilization was lower with CTAC. However, superior refolding efficiency resulted in final yields of purified rPGH being in the order of CTAC greater than urea greater than or equal to guanidinium chloride. Detailed comparison of the different rPGH preparations as well as pituitary-derived growth hormone by h.p.l.c., native PAGE, c.d. spectral analysis and radioreceptor-binding assay showed that the CTAC-derived rPGH was essentially indistinguishable from the urea and guanidinium chloride preparations. The CTAC-derived rPGH was of greater biopotency than pituitary-derived growth hormone. The advantages of CTAC over urea and guanidinium chloride for increasing recovery of monomeric rPGH by minimizing aggregation during refolding in vitro were also found with recombinant sheep interleukin-I beta and a sheep insulin-like growth factor II fusion protein. In addition, the bioactivity of the CTAC-derived recombinant interleukin-1 beta was approximately ten-fold greater than that of an equivalent amount obtained from urea and guanidinium chloride preparations. It is concluded that CTAC represents, in general, an excellent additional approach or a superior alternative to urea and in particular guanidinium chloride for solubilization and recovery of bioactive recombinant proteins from inclusion bodies. Images Fig. 1. Fig. 3. Fig. 5. Fig. 9. PMID:1497625

  5. Replication of Boid Inclusion Body Disease-Associated Arenaviruses Is Temperature Sensitive in both Boid and Mammalian Cells

    PubMed Central

    Kipar, Anja; Korzyukov, Yegor; Bell-Sakyi, Lesley; Vapalahti, Olli; Hetzel, Udo

    2014-01-01

    ABSTRACT Boid inclusion body disease (BIDB) is a fatal disease of boid snakes, the etiology of which has only recently been revealed following the identification of several novel arenaviruses in diseased snakes. BIBD-associated arenaviruses (BIBDAV) are genetically divergent from the classical Old and New World arenaviruses and also differ substantially from each other. Even though there is convincing evidence that BIBDAV are indeed the etiological agent of BIBD, the BIBDAV reservoir hosts—if any exist besides boid snakes themselves—are not yet known. In this report, we use University of Helsinki virus (UHV; a virus that we isolated from a Boa constrictor with BIBD) to show that BIBDAV can also replicate effectively in mammalian cells, including human cells, provided they are cultured at 30°C. The infection induces the formation of cytoplasmic inclusion bodies (IB), comprised mainly of viral nucleoprotein (NP), similar to those observed in BIBD and in boid cell cultures. Transferring infected cells from 30°C to 37°C ambient temperature resulted in progressive declines in IB formation and in the amounts of viral NP and RNA, suggesting that BIBDAV growth is limited at 37°C. These observations indirectly indicate that IB formation is linked to viral replication. In addition to mammalian and reptilian cells, UHV infected arthropod (tick) cells when grown at 30°C. Even though our findings suggest that BIBDAV have a high potential to cross the species barrier, their inefficient growth at mammalian body temperatures indicates that the reservoir hosts of BIBDAV are likely species with a lower body temperature, such as snakes. IMPORTANCE The newly discovered boid inclusion body disease-associated arenaviruses (BIBDAV) of reptiles have drastically altered the phylogeny of the family Arenavirus. Prior to their discovery, known arenaviruses were considered mainly rodent-borne viruses, with each arenavirus species having its own reservoir host. BIBDAV have so far been

  6. Case report: inclusion body disease of cranes: a serological follow-up to the 1978 die-off

    USGS Publications Warehouse

    Docherty, D.E.; Romaine, Renee I.

    1983-01-01

    A herpesvirus was isolated from captive cranes involved in a 1978 die-off. Neutralizing antibody to this virus was detected in this captive population as early as 1975 and consistently thereafter through 1979. Exposure to the virus evidently occurred at least 2 1/2 years before the die-off, without causing any mortality diagnosed as being caused by inclusion body disease of cranes (IBDC). Overcrowding and environmental conditions in 1978 may have contributed to the deaths of certain species of cranes in one area and not in another. Mortality ratios and serological data suggest that crane species vary in their response to IBDC virus.

  7. The receptor for advanced glycation end products promotes bacterial growth at distant body sites in Staphylococcus aureus skin infection.

    PubMed

    Achouiti, Ahmed; Van't Veer, Cornelis; de Vos, Alex F; van der Poll, Tom

    2015-09-01

    The receptor for advanced glycation endproducts (RAGE) has been implicated in the regulation of skin inflammation. We here sought to study the role of RAGE in host defense during skin infection caused by Staphylococcus (S.) aureus, the most common pathogen in this condition. Wild-type (Wt) and RAGE deficient (rage(-/-)) mice were infected subcutaneously with S. aureus and bacterial loads and local inflammation were quantified at regular intervals up to 8 days after infection. While bacterial burdens were similar in both mouse strains at the primary site of infection, rage(-/-) mice had lower bacterial counts in lungs and liver. Skin cytokine and chemokine levels did not differ between groups. In accordance with the skin model, direct intravenous infection with S. aureus was associated with lower bacterial loads in lungs and liver of rage(-/-) mice. Together these data suggest that RAGE does not impact local host defense during S. aureus skin infection, but facilitates bacterial growth at distant body sites. PMID:26086798

  8. Procalcitonin as a predictive biomarker for total body irradiation induced bacterial load and lethality in mice

    PubMed Central

    Biju, Prabath G.; Garg, Sarita; Wang, Wenze; Choudhry, Mashkoor A.; Kovacs, Elizabeth J.; Fink, Louis M.; Hauer-Jensen, Martin

    2012-01-01

    Sepsis is the leading cause of mortality in intensive care units. Early detection and intervention are critical to prevent death. The acute radiation syndrome is characterized by damage of the gastrointestinal and hematopoietic systems. Translocation of intestinal microflora combined with immune system compromise may lead to septicemia and death. This work examined the utility of procalcitonin, a clinical sepsis biomarker, in a mouse model of radiation toxicity. C57/BL6 mice were exposed to total body irradiation (TBI). Intestinal mucosal permeability was measured in vivo, and liver bacterial load and plasma levels of procalcitonin (PCT), lipopolysaccharide (LPS), and lipopolysaccharide binding protein (LBP) were measured at baseline and 3.5, 7, and 10 days after TBI. The value of early PCT in predicting subsequent lethality was determined by receiver operator characteristics (ROC) analysis. Four days after TBI a dose-dependent increase in permeability of the intestinal mucosa was observed, while bacterial translocation was present from day 7 onward. There was a high positive correlation between bacterial translocation and all sepsis biomarkers, with PCT exhibiting the strongest correlation. Moreover, plasma PCT levels were elevated already from day 3.5 onwards, whereas, LPS was elevated from day 7 and LBP only 10 days after TBI. ROC analysis revealed that PCT levels measured 3.5 days after TBI predicted lethality at 10 days. These data demonstrate the value of PCT as an early biomarker in radiation-induced bacteremia for mouse studies and suggest that clinical results from other septic conditions may apply to post-radiation septicemia in humans. PMID:22576002

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

    PubMed Central

    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

    2015-01-01

    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 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. PMID:25791609

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

    PubMed

    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

    2015-01-01

    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 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. PMID:25791609

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

    PubMed Central

    Huang, Q.; Su, Y. B.; Li, D. F.; Liu, L.; Huang, C. F.; Zhu, Z. P.; Lai, C. H.

    2015-01-01

    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

  12. Highly Effective Renaturation of a Streptokinase from Streptococcus pyogenes DT7 as Inclusion Bodies Overexpressed in Escherichia coli

    PubMed Central

    Nguyen, Sy Le Thanh; Quyen, Dinh Thi; Vu, Hong Diep

    2014-01-01

    The streptokinase (SK) is emerging as an important thrombolytic therapy agent in the treatment of patients suffering from cardiovascular diseases. We reported highly effective renaturation of a SK from S. pyogeness DT7 overexpressed in E. coli, purification, and biochemical characterization. A gene coding for the SK was cloned from S. pyogeness DT7. Because accumulation of active SK is toxic to the host cells, we have expressed it in the form of inclusion bodies. The mature protein was overexpressed in E. coli BL21 DE3/pESK under the control of the strong promoter tac induced by IPTG with a level of 60% of the total cell proteins. The activity of the rSK, renatured in phosphate buffer supplemented with Triton X-100 and glycerol, was covered with up to 41 folds of its initial activity. The purified of protein was identified with MALDI-TOF mass spectrometry through four peptide fragments, which showed 100% identification to the corresponding peptides of the putative SK from GenBank. Due to overexpression and highly effective renaturation of large amounts of inclusion bodies, the recombinant E. coli BL21 DE3/pESK system could be potentially applied for large-scale production of SK used in the therapy of acute myocardial infarction. PMID:24883307

  13. Integrated continuous dissolution, refolding and tag removal of fusion proteins from inclusion bodies in a tubular reactor.

    PubMed

    Pan, Siqi; Zelger, Monika; Jungbauer, Alois; Hahn, Rainer

    2014-09-20

    An integrated continuous tubular reactor system was developed for processing an autoprotease expressed as inclusion bodies. The inclusion bodies were suspended and fed into the tubular reactor system for continuous dissolving, refolding and precipitation. During refolding, the dissolved autoprotease cleaves itself, separating the fusion tag from the target peptide. Subsequently, the cleaved fusion tag and any uncleaved autoprotease were precipitated out in the precipitation step. The processed exiting solution results in the purified soluble target peptide. Refolding and precipitation yields performed in the tubular reactor were similar to batch reactor and process was stable for at least 20 h. The authenticity of purified peptide was also verified by mass spectroscopy. Productivity (in mg/l/h and mg/h) calculated in the tubular process was twice and 1.5 times of the batch process, respectively. Although it is more complex to setup a tubular than a batch reactor, it offers faster mixing, higher productivity and better integration to other bioprocessing steps. With increasing interest of integrated continuous biomanufacturing, the use of tubular reactors in industrial settings offers clear advantages. PMID:24950296

  14. Studies of a nuclear matrix protein restricted to normal brain cells and lead-induced intranuclear inclusion bodies of kidney

    SciTech Connect

    Shelton, K.; Egle, P.; Redford, K.; Bigbee, J.

    1986-05-01

    A nuclear matrix protein, p32/6.3, with an unusual tissue distribution, has been identified. Protein from 21 tissues was surveyed by immunoprobing Western blots. In normal adult rats p32/6.3 is found only in grey matter from the cerebrum and the cerebellum, occurring in both neurons and astrocytes. Other brain cell types have not been examined. The protein appears to be developmentally regulated. It is detectable in the brain within a few days after birth and reaches adult levels within one to two weeks. Brain p32/6.3 has been found in all animals tested including rat, mouse, dog, cow, pig, chicken and human. This conservation indicates a fundamental role for p32/6.3 in the nucleus of brain cells. Possible functions for p32/6.3 may be indicated by a second novel occurrence. Chronic lead poisoning characteristically induces intranuclear inclusion bodies in the cells lining kidney proximal tubules. p32/6.3 is a major constituent of these inclusion bodies. They are also rich in lead and other metals including calcium, iron, zinc, copper and cadmium. These diverse observations suggest that p32/6.3 may have a role in metal homeostasis in the brain of normal animals.

  15. Interferon-alpha 2b quantification in inclusion bodies using reversed phase-ultra performance liquid chromatography (RP-UPLC).

    PubMed

    Cueto-Rojas, H F; Pérez, N O; Pérez-Sánchez, G; Ocampo-Juárez, I; Medina-Rivero, E

    2010-04-15

    Interferon-alpha 2b (IFN-alpha 2b) is a recombinant therapeutic cytokine produced as inclusion bodies using a strain of Escherichia coli as expression system. After fermentation and recovery, it is necessary to know the amount of recombinant IFN-alpha 2b, in order to determine the yield and the load for solubilization, and chromatographic protein purification steps. The present work details the validation of a new short run-time and fast sample-preparation method to quantify IFN-alpha 2b in inclusion bodies using Reversed Phase-Ultra Performance Liquid Chromatography (RP-UPLC). The developed method demonstrated an accuracy of 100.28%; the relative standard deviations for method precision, repeatability and inter-day precision tests were found to be 0.57%, 1.54% and 1.83%, respectively. Linearity of the method was assessed in the range of concentrations from 0.05 mg/mL to 0.5 mg/mL, the curve obtained had a determination coefficient (r(2)) of 0.9989. Detection and quantification limits were found to be 0.008 mg/mL and 0.025 mg/mL, respectively. The method also demonstrated robustness for changes in column temperature, and specificity against host proteins and other recombinant protein expressed in the same E. coli strain. PMID:20299292

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

    PubMed Central

    Nowakowska, Justyna; Landmann, Regine; Khanna, Nina

    2014-01-01

    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. PMID:27025752

  17. Comparison of bacterial and archaeal communities in depth-resolved zones in an LNAPL body.

    PubMed

    Irianni-Renno, Maria; Akhbari, Daria; Olson, Mitchell R; Byrne, Adam P; Lefèvre, Emilie; Zimbron, Julio; Lyverse, Mark; Sale, Thomas C; De Long, Susan K

    2016-04-01

    Advances in our understanding of the microbial ecology at sites impacted by light non-aqueous phase liquids (LNAPLs) are needed to drive development of optimized bioremediation technologies, support longevity models, and develop culture-independent molecular tools. In this study, depth-resolved characterization of geochemical parameters and microbial communities was conducted for a shallow hydrocarbon-impacted aquifer. Four distinct zones were identified based on microbial community structure and geochemical data: (i) an aerobic, low-contaminant mass zone at the top of the vadose zone; (ii) a moderate to high-contaminant mass, low-oxygen to anaerobic transition zone in the middle of the vadose zone; (iii) an anaerobic, high-contaminant mass zone spanning the bottom of the vadose zone and saturated zone; and (iv) an anaerobic, low-contaminant mass zone below the LNAPL body. Evidence suggested that hydrocarbon degradation is mediated by syntrophic fermenters and methanogens in zone III. Upward flux of methane likely contributes to promoting anaerobic conditions in zone II by limiting downward flux of oxygen as methane and oxygen fronts converge at the top of this zone. Observed sulfate gradients and microbial communities suggested that sulfate reduction and methanogenesis both contribute to hydrocarbon degradation in zone IV. Pyrosequencing revealed that Syntrophus- and Methanosaeta-related species dominate bacterial and archaeal communities, respectively, in the LNAPL body below the water table. Observed phylotypes were linked with in situ anaerobic hydrocarbon degradation in LNAPL-impacted soils. PMID:26691516

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

    SciTech Connect

    Lovato, Alessandro; Gandolfi, Stefano; Carlson, Joseph; Pieper, S. C.; Schiavilla, Rocco

    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.

  19. [Lamellar Inclusion Bodies in the Retinal Pigment Epithelium of Diurnal Rodents].

    PubMed

    Samosudova, N V; Orlov, O U; Golyshev, S A

    2015-01-01

    The ultrastructure of the retinal pigment epithelium of a diurnal rodent (Brandt's vole) was described taking into account 1) the functions of the pigment epithelium as a participant in the renewal of photoreceptor outer segment and. 2) digestion of outer segment membranes into phagosomes of the retinal pigment epithelium. The myeloid bodies were observed after exposure of the pigment epithelium to light (200 lux, 4 hours) and darkness (0,1 lux, 1,5-hour). In the cytoplasm of the pigment epithelium of the vole no myeloid bodies were observed. Instead of it small lamellar bodies, which have the spiral form and size (from - 200 to 400 nm) were found. The structure of these lamellar bodies was described. Furthermore, the structures, which were presumably responsible for the transport of the digested material, were revealed. The evidence of it is the presence of 1) dense precipitate in the apical domain of the pigment epithelium and 2) microtubules which participate in transport of this precipitate. PMID:26591606

  20. Body Temperature at the Emergency Department as a Predictor of Mortality in Patients With Bacterial Infection

    PubMed Central

    Yamamoto, Shungo; Yamazaki, Shin; Shimizu, Tsunehiro; Takeshima, Taro; Fukuma, Shingo; Yamamoto, Yosuke; Tochitani, Kentaro; Tsuchido, Yasuhiro; Shinohara, Koh; Fukuhara, Shunichi

    2016-01-01

    Abstract Hypothermia is a risk factor for death in intensive care unit (ICU) patients with severe sepsis and septic shock. In the present study, we investigated the association between body temperature (BT) on arrival at the emergency department (ED) and mortality in patients with bacterial infection. We conducted a retrospective cohort study in consecutive ED patients over 15 years of age with bacterial infection who were admitted to an urban teaching hospital in Japan between 2010 and 2012. The main outcome measure was 30-day in-hospital mortality. Each patient was assigned to 1 of 6 categories based on BT at ED admission. We conducted multivariable logistic regression analysis to adjust for predictors of death. A total of 913 patients were enrolled in the study. The BT categories were <36, 36 to 36.9, 37 to 37.9, 38 to 38.9, 39 to 39.9, and ≥40 °C, with respective mortalities of 32.5%, 14.1%, 8.7%, 8.2%, 5.7%, and 5.3%. Multivariable analysis showed that the risk of death was significantly low in patients with BT 37 to 37.9 °C (adjusted odds ratio [AOR]: 0.2; 95% confidence interval [CI] 0.1–0.6, P = 0.003), 38–38.9 °C (AOR: 0.2; 95% CI 0.1–0.6, P = 0.002), 39–39.9 °C (AOR: 0.2; 95% CI 0.1–0.5, P = 0.001), and ≥40 °C (AOR: 0.1; 95% CI 0.02–0.4, P = 0.001), compared with hypothermic patients (BT <36 °C). The higher BT on arrival at ED, the better the outcomes observed in patients with bacterial infection were. PMID:27227924

  1. 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

    PubMed Central

    Chang, Li-Wen; Fu, Ann; Wozniak, Edward; Chow, Marjorie; Duke, Diane G.; Green, Linda; Kelley, Karen; Hernandez, Jorge A.; Jacobson, Elliott R.

    2013-01-01

    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. PMID:24340066

  2. 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.

    PubMed

    Chang, Li-Wen; Fu, Ann; Wozniak, Edward; Chow, Marjorie; Duke, Diane G; Green, Linda; Kelley, Karen; Hernandez, Jorge A; Jacobson, Elliott R

    2013-01-01

    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. PMID:24340066

  3. Systematic Significance of Cell Inclusions in Haemodoraceae and Allied Families: Silica Bodies and Tapetal Raphides

    PubMed Central

    PRYCHID, CHRISTINA J.; FURNESS, CAROL A.; RUDALL, PAULA J.

    2003-01-01

    This paper presents the first record of silica deposits in tissues of Haemodoraceae and adds new records of tapetal raphides in this family. Within the order Commelinales, silica is present in leaves of three families (Hanguanacaeae, Haemodoraceae and Commelinaceae), but entirely absent from the other two (Pontederiaceae and Philydraceae). Presence or absence of characteristic cell inclusions may have systematic potential in commelinid monocotyledons, although the existing topology indicates de novo gains and losses in individual families. Silica sand was observed in leaves of five out of nine genera examined of Haemodoraceae, predominantly in vascular bundle sheath cells and epidermal cells. Within Haemodoraceae, silica is limited to subfamily Conostylidoideae. The occurrence of silica in Phlebocarya supports an earlier transfer of this genus from Haemodoroideae to Conostylidoideae. The presence of raphides (calcium oxalate crystals) in the anther tapetum represents a rare character, only reported in a few monocot families of the order Commelinales, and possibly representing a mechanism for regulation of cytoplasmic free calcium levels. Tapetal raphides were observed here in Anigozanthus and Conostylis (both Haemodoraceae), and Tradescantia (Commelinaceae), thus supplementing two earlier records in Haemodoraceae, Philydraceae and Commelinaceae. PMID:14507742

  4. Iron and aluminum deposition in the meninges of the lamprey: identification of an aluminum-ferritin inclusion body

    SciTech Connect

    Youson, J.H.; Sargent, P.A.; Pearce, G.W.

    1989-01-01

    The meningeal tissue of the brain and spinal cord of larval and juvenile adults of lampreys (Petromyzon marinus) was examined by routine electron microscopy, electron microscopic histochemistry, and electron-probe x-ray microanalysis to locate sites of iron deposition. A magnetometer was used for identification of ferromagnetic iron. Ferritin particles, representing ferric iron, are present in abundance within the cytoplasmic matrices and in dense bodies of meningeal cells of both the brain and spinal cord of larvae and juveniles. These round cells of the meninges also contain abundant glycogen and lipid. Small quantities of ferrous iron are associated to the latter inclusion. Aluminum deposits are present within an electron-dense material of many ferritin-containing inclusions of meningeal cells of the larval brain. Ferromagnetic material was not detected in larval and upstream-migrant lampreys. The deposition of iron and aluminum in the meninges of lampreys may be related to physiological and environmental factors, respectively, and/or to an important interaction between the two metals.

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

    NASA Astrophysics Data System (ADS)

    Morris, Titus; Bogner, Scott

    2015-10-01

    The In-Medium Similarity Renormalization Group (IM-SRG) has been applied successfully not only to several closed shell finite nuclei, but has recently been used to produce effective shell model interactions that are competitive with phenomenological interactions in the SD shell. A recent alternative method for solving of the IM-SRG equations, called 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.

  6. Purification and characterization of naturally occurring HIV-1 (South African subtype C) protease mutants from inclusion bodies.

    PubMed

    Maseko, Sibusiso B; Natarajan, Satheesh; Sharma, Vikas; Bhattacharyya, Neelakshi; Govender, Thavendran; Sayed, Yasien; Maguire, Glenn E M; Lin, Johnson; Kruger, Hendrik G

    2016-06-01

    Human immunodeficiency virus (HIV) infections in sub-Saharan Africa represent about 56% of global infections. Many studies have targeted HIV-1 protease for the development of drugs against AIDS. Recombinant HIV-1 protease is used to screen new drugs from synthetic compounds or natural substances. Along with the wild type (C-SA) we also over-expressed and characterized two mutant forms from patients that had shown resistance to protease inhibitors. Using recombinant DNA technology, we constructed three recombinant plasmids in pGEX-6P-1 and expressed them containing a sequence encoding wild type HIV protease and two mutants (I36T↑T contains 100 amino acids and L38L↑N↑L contains 101 amino acids). These recombinant proteins were isolated from inclusion bodies by using QFF anion exchange and GST trap columns. In SDS-PAGE, we obtained these HIV proteases as single bands of approximately 11.5, 11.6 and 11.7 kDa for the wild type, I36T↑Tand L38L↑N↑L mutants, respectively. The enzyme was recovered efficiently (0.25 mg protein/L of Escherichia coli culture) and had high specific activity of 2.02, 2.20 and 1.33 μmol min(-1) mg(-1) at an optimal pH of 5 and temperature of 37 °C for the wild type, I36T↑T and L38L↑N↑L, respectively. The method employed here provides an easy and rapid purification of the HIV-1(C-SA) protease from the inclusion bodies, with high yield and high specific activities. PMID:26917227

  7. Affect of corn germ meal inclusion in pig diets bacterial ecology in the cecum and proximal colon

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inclusion of bio-fuel co-products in swine diets is becoming more common due to greater availability and increasing cereal grain costs. These co-products have lower starch and higher fiber concentrations. Twenty-four pigs were adapted to diets with either corn or solvent extracted corn germ meal (...

  8. Refolding and purification of Bothropstoxin-I, a Lys49-phospholipase A2 homologue, expressed as inclusion bodies in Escherichia coli.

    PubMed

    Ward, R J; de Oliveira, A H; Bortoleto, R K; Rosa, J C; Faça, V M; Greene, L J

    2001-02-01

    Hydrolysis of phospholipids by Group II phospholipase A2 enzymes involves a nucleophilic attack on the sn-2 ester bond by the His48 residue and stabilization of the reaction intermediate by a Ca2+ ion cofactor bound to the Asp49 residue in the protein active site region. Bothropstoxin-I (BthTX-I) is a PLA(2) variant present in the venom of the snake Bothrops jararacussu which shows a Asp49 to Lys substitution and which lacks hydrolytic activity yet damages artificial membranes by a noncatalytic Ca2+-independent mechanism. In order to better characterize this unusual mechanism of membrane damage, we have established an expression system for BthTX-I in Escherichia coli. The DNA-coding sequence for BthTX-I was subcloned into the vector pET11-d, and the BthTX-I was expressed as inclusion bodies in E. coli BL21(DE3). The native BthTX-I contains seven disulfide bonds, and a straightforward protocol has been developed to refold the recombinant protein at high protein concentration in the presence of surfactants using a size-exclusion chromatography matrix. After refolding, recovery yields of 2.5% (corresponding to 4-5 mg of refolded recombinant BthTX-I per liter of bacterial culture) were routinely obtained. After refolding, identical fluorescent and circular dichroism spectra were obtained for the recombinant BthTX-I compared to those of the native protein. Furthermore, the native and refolded recombinant protein demonstrated identical membrane-damaging properties as evaluated by measuring the release of an entrapped fluorescent marker from liposomes. PMID:11162398

  9. The many-body problem for anisotropic membrane inclusions and the self-assembly of "saddle" defects into an "egg carton".

    PubMed

    Dommersnes, Paul G; Fournier, Jean-Baptiste

    2002-12-01

    We calculate the many-body, nonpairwise interaction between N rigid, anisotropic membrane inclusions by modeling them as point-like constraints on the membrane's curvature tensor and by minimizing the membrane's curvature energy. Because multipolar distortions of higher-order decay on very short distances, our calculation gives the correct elastic interaction energy for inclusions separated by distances of the order of several times their size. As an application, we show by thermally equilibrating the many-body elastic energy using a Monte Carlo algorithm, that inclusions shaped as "saddles" attract each other and build an "egg-carton" structure. The latter is reminiscent of some patterns observed in membranes obtained from biological extracts, the origin of which is still mysterious. PMID:12496066

  10. Mutant LRRK2 Toxicity in Neurons Depends on LRRK2 Levels and Synuclein But Not Kinase Activity or Inclusion Bodies

    PubMed Central

    Skibinski, Gaia; Nakamura, Ken; Cookson, Mark R.

    2014-01-01

    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. PMID:24403142

  11. Hereditary inclusion body myopathy: A myopathy with unique topography of weakness, yet frequently misdiagnosed: Case series and review of literature

    PubMed Central

    Das, Biplab; Goyal, Manoj Kumar; Bhatkar, Sanat Ramchandra; Vinny, Pulikottil Wilson; Modi, Manish; Lal, Vivek; Gayathri, N.; Mahadevan, Anitha; Radotra, Bishan Dass

    2016-01-01

    Background: Hereditary inclusion body myopathy (HIBM) continues to be underrecognized clinically despite a characteristic topography of weakness with total sparing of quadriceps muscles and patient being wheelchair bound. We report seven patients of HIBM from four families in North India. Methods and Results: Seven patients from four different families were diagnosed to have HIBM. There was no consanguinity in any of the families. While one patient had two affected siblings, another had one affected siblings and the family history was noncontributory in two patients. Two of the siblings were available for examination and confirmed clinically to be suffering from HIBM. Among the seven patients, only one was still ambulatory at the time of diagnosis. Discussion: This is the first case report of occurrence of HIBM in North Indian population. Despite its unique clinical presentation, HIBM is frequently misdiagnosed resulting in unnecessary diagnostic and therapeutic interventions. A high index of suspicion of this rare myopathy along with proper clinical examination may go a long way in accurate prognostication and management of these patients. PMID:27011643

  12. Refolding and purification of recombinant L-asparaginase from inclusion bodies of E. coli into active tetrameric protein.

    PubMed

    Upadhyay, Arun K; Singh, Anupam; Mukherjee, K J; Panda, Amulya K

    2014-01-01

    A tetrameric protein of therapeutic importance, Escherichia coli L-asparaginase-II was expressed in Escherichia coli as inclusion bodies (IBs). Asparaginase IBs were solubilized using low concentration of urea and refolded into active tetrameric protein using pulsatile dilution method. Refolded asparaginase was purified in two steps by ion-exchange and gel filtration chromatographic techniques. The recovery of bioactive asparaginase from IBs was around 50%. The melting temperature (Tm) of the purified asparaginase was found to be 64°C. The specific activity of refolded, purified asparaginase was found to be comparable to the commercial asparaginase (190 IU/mg). Enzymatic activity of the refolded asparaginase was high even at four molar urea solutions, where the IB aggregates are completely solubilized. From the comparison of chemical denaturation data and activity at different concentrations of guanidine hydrochloride, it was observed that dissociation of monomeric units precedes the complete loss of helical secondary structures. Protection of the existing native-like protein structure during solubilization of IB aggregates with 4 M urea improved the propensity of monomer units to form oligomeric structure. Our mild solubilization technique retaining native-like structures, improved recovery of asparaginase in bioactive tetrameric form. PMID:25309524

  13. Refolding and purification of recombinant L-asparaginase from inclusion bodies of E. coli into active tetrameric protein

    PubMed Central

    Upadhyay, Arun K.; Singh, Anupam; Mukherjee, K. J.; Panda, Amulya K.

    2014-01-01

    A tetrameric protein of therapeutic importance, Escherichia coli L-asparaginase-II was expressed in Escherichia coli as inclusion bodies (IBs). Asparaginase IBs were solubilized using low concentration of urea and refolded into active tetrameric protein using pulsatile dilution method. Refolded asparaginase was purified in two steps by ion-exchange and gel filtration chromatographic techniques. The recovery of bioactive asparaginase from IBs was around 50%. The melting temperature (Tm) of the purified asparaginase was found to be 64°C. The specific activity of refolded, purified asparaginase was found to be comparable to the commercial asparaginase (190 IU/mg). Enzymatic activity of the refolded asparaginase was high even at four molar urea solutions, where the IB aggregates are completely solubilized. From the comparison of chemical denaturation data and activity at different concentrations of guanidine hydrochloride, it was observed that dissociation of monomeric units precedes the complete loss of helical secondary structures. Protection of the existing native-like protein structure during solubilization of IB aggregates with 4 M urea improved the propensity of monomer units to form oligomeric structure. Our mild solubilization technique retaining native-like structures, improved recovery of asparaginase in bioactive tetrameric form. PMID:25309524

  14. Formation of Mallory Body-like Inclusions and Cell Death Induced by Deregulated Expression of Keratin 18

    PubMed Central

    Nakamichi, Ikuo; Hatakeyama, Shigetsugu; Nakayama, Keiichi I.

    2002-01-01

    Mallory bodies (MBs) are cytoplasmic inclusions that contain keratin 8 (K8) and K18 and are present in hepatocytes of individuals with alcoholic liver disease, nonalcoholic steatohepatitis, or benign or malignant hepatocellular neoplasia. Mice fed long term with griseofulvin are an animal model of MB formation. However, the lack of a cellular model has impeded understanding of the molecular mechanism of this process. Culture of HepG2 cells with griseofulvin has now been shown to induce both the formation of intracellular aggregates containing K18 as well as an increase in the abundance of K18 mRNA. Overexpression of K18 in HepG2, HeLa, or COS-7 cells also induced the formation of intracellular aggregates that stained with antibodies to ubiquitin and with rhodamine B (characteristics of MBs formed in vivo), eventually leading to cell death. The MB-like aggregates were deposited around centrosomes and disrupted the microtubular array. Coexpression of K8 with K18 restored the normal fibrous pattern of keratin distribution and reduced the toxicity of K18. In contrast, an NH2-terminal deletion mutant of K8 promoted the formation of intracellular aggregates even in the absence of K18 overexpression. Deregulated expression of K18, or an imbalance between K8 and K18, may thus be an important determinant of MB formation, which compromises the function of centrosomes and the microtubule network and leads to cell death. PMID:12388748

  15. Atypical sporadic CJD-MM phenotype with white matter kuru plaques associated with intranuclear inclusion body and argyrophilic grain disease.

    PubMed

    Berghoff, Anna S; Trummert, Anita; Unterberger, Ursula; Ströbel, Thomas; Hortobágyi, Tibor; Kovacs, Gabor G

    2015-08-01

    We describe an atypical neuropathological phenotype of sporadic Creutzfeldt-Jakob disease in a 76-year-old man. The clinical symptoms were characterized by progressive dementia, gait ataxia, rigidity and urinary incontinence. The disease duration was 6 weeks. MRI did not show prominent atrophy or hyperintensities in cortical areas, striatum or thalamus. Biomarker examination of the cerebrospinal fluid deviated from that seen in pure Alzheimer's disease. Triphasic waves in the EEG were detected only later in the disease course, while 14-3-3 assay was positive. PRNP genotyping revealed methionine homozygosity (MM) at codon 129. Neuropathology showed classical CJD changes corresponding to the MM type 1 cases. However, a striking feature was the presence of abundant kuru-type plaques in the white matter. This rare morphology was associated with neuropathological signs of intranuclear inclusion body disease and advanced stage of argyrophilic grain disease. These alterations did not show correlation with each other, thus seemed to develop independently. This case further highlights the complexity of neuropathological alterations in the ageing brain. PMID:25783686

  16. Aquareovirus NS80 Initiates Efficient Viral Replication by Retaining Core Proteins within Replication-Associated Viral Inclusion Bodies

    PubMed Central

    Yan, Liming; Zhang, Jie; Guo, Hong; Yan, Shicui; Chen, Qingxiu; Zhang, Fuxian; Fang, Qin

    2015-01-01

    Viral inclusion bodies (VIBs) are specific intracellular compartments for reoviruses replication and assembly. Aquareovirus nonstructural protein NS80 has been identified to be the major constituent for forming globular VIBs in our previous study. In this study, we investigated the role of NS80 in viral structural proteins expression and viral replication. Immunofluorescence assays showed that NS80 could retain five core proteins or inner-capsid proteins (VP1-VP4 and VP6), but not outer-capsid proteins (VP5 and VP7), within VIBs in co-transfected or infected cells. Further co-immunoprecipitation analysis confirmed that NS80 could interact with each core protein respectively. In addition, we found that newly synthesized viral RNAs co-localized with VIBs. Furthermore, time-course analysis of viral structural proteins expression showed that the expression of NS80 was detected first, followed by the detection of inner shell protein VP3, and then of other inner-capsid proteins, suggesting that VIBs were essential for the formation of viral core frame or progeny virion. Moreover, knockdown of NS80 by shRNA not only inhibited the expression of aquareovirus structural proteins, but also inhibited viral infection. These results indicated that NS80-based VIBs were formed at earlier stage of infection, and NS80 was able to coordinate the expression of viral structural proteins and viral replication. PMID:25938226

  17. A Brazilian family with hereditary inclusion body myopathy associated with Paget disease of bone and frontotemporal dementia.

    PubMed

    Fanganiello, R D; Kimonis, V E; Côrte, C C; Nitrini, R; Passos-Bueno, M R

    2011-04-01

    Inclusion body myopathy associated with Paget disease and frontotemporal dementia (IBMPFD) is a progressive and usually misdiagnosed autosomal dominant disorder. It is clinically characterized by a triad of features: proximal and distal myopathy, early onset Paget disease of bone (PDB), and frontotemporal dementia (FTD). It is caused by missense mutations in the valosin-containing protein (VCP) gene. We describe here the clinical and molecular findings of the first Brazilian family identified with IBMPFD. Progressive myopathy affecting the limb girdles was detected by clinical examination followed by muscle biopsy and creatine kinase measurement. PDB was suggested after anatomopathological bone examination and FTD was diagnosed by clinical, neuropsychological and language evaluations. Brain magnetic resonance revealed severe atrophy of the anterior temporal lobes, including the hippocampi. A R93C mutation in VCP was detected by direct sequencing screening in subject W (age 62) and in his mother. Four more individuals diagnosed with "dementia" were reported in this family. We also present a comprehensive genotype-phenotype correlation analysis of mutations in VCP in 182 patients from 29 families described in the literature and show that while IBM is a conspicuously penetrant symptom, PDB has a lower penetrance when associated with mutations in the AAAD1 domain and FTD has a lower penetrance when associated with mutations in the Junction (L1-D1) domain. Furthermore, the R93C mutation is likely to be associated with the penetrance of all the clinical symptoms of the triad. PMID:21412659

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

    PubMed

    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. PMID:26363119

  19. Exploring the Medicinal Potential of the Fruit Bodies of Oyster Mushroom, Pleurotus ostreatus (Agaricomycetes), against Multidrug-Resistant Bacterial Isolates.

    PubMed

    Skariyachan, Sinosh; Prasanna, Apoorva; Manjunath, Sirisha P; Karanth, Soujanya S; Nazre, Ambika

    2016-01-01

    Bacterial resistance to present-generation antibiotics is increasing drastically, which has become a major public health concern. The present study focuses on demonstrating the antimicrobial potential of fruit bodies of the culinary/medicinal oyster mushroom Pleurotus ostreatus against clinical pathogens. Five bacterial isolates were collected from Sagar Hospital in Bangalore, India. The collected strains were grown on selective and differential media and antibiotic susceptibility testing was applied using 48 antibiotics by disc diffusion assay. The antibacterial efficiency of the mushroom extract against clinical pathogens, which were found to be multidrug resistant (MDR) to most of the tested antibiotics, was studied. The yield of cultivated mushrooms was evident at moist, cooler, and humid conditions. The clinical isolates of Staphylococcus aureus, Salmonella typhi, Acinetobacter sp., Proteus mirabilis, and Proteus spp. were found to be MDR to β-lactam, fluoroquinolones, sulfonamides, third- and fourth-generation cephalosporins, aminoglycosides, macrolides, tetracyclines, and carbapenems. The methanolic extracts of mushroom fruit bodies were found to be more effective than present-generation antibiotics against methicillin- and vancomycin- resistant S. aureus, S. typhi, Acinetobacter sp., and P. mirabilis at a concentration ranging from 50 to 100 µg/disc or 50 to 100 µL/well. The current study suggests that the methanolic extract of P. ostreatus can be used as a promising antibacterial agent against MDR bacterial pathogens. PMID:27481158

  20. The RNA-binding motif 45 (RBM45) protein accumulates in inclusion bodies in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP) patients.

    PubMed

    Collins, Mahlon; Riascos, David; Kovalik, Tina; An, Jiyan; Krupa, Kelly; Krupa, Kristin; Hood, Brian L; Conrads, Thomas P; Renton, Alan E; Traynor, Bryan J; Bowser, Robert

    2012-11-01

    RNA-binding protein pathology now represents one of the best characterized pathologic features of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration patients with TDP-43 or FUS pathology (FTLD-TDP and FTLD-FUS). Using liquid chromatography tandem mass spectrometry, we identified altered levels of the RNA-binding motif 45 (RBM45) protein in the cerebrospinal fluid (CSF) of ALS patients. This protein contains sequence similarities to TAR DNA-binding protein 43 (TDP-43) and fused-in-sarcoma (FUS) that are contained in cytoplasmic inclusions of ALS and FTLD-TDP or FTLD-FUS patients. To further characterize RBM45, we first verified the presence of RBM45 in CSF and spinal cord tissue extracts of ALS patients by immunoblot. We next used immunohistochemistry to examine the subcellular distribution of RBM45 and observed in a punctate staining pattern within nuclei of neurons and glia in the brain and spinal cord. We also detected RBM45 cytoplasmic inclusions in 91 % of ALS, 100 % of FTLD-TDP and 75 % of Alzheimer's disease (AD) cases. The most extensive RBM45 pathology was observed in patients that harbor the C9ORF72 hexanucleotide repeat expansion. These RBM45 inclusions were observed in spinal cord motor neurons, glia and neurons of the dentate gyrus. By confocal microscopy, RBM45 co-localizes with ubiquitin and TDP-43 in inclusion bodies. In neurons containing RBM45 cytoplasmic inclusions we often detected the protein in a punctate pattern within the nucleus that lacked either TDP-43 or ubiquitin. We identified RBM45 using a proteomic screen of CSF from ALS and control subjects for candidate biomarkers, and link this RNA-binding protein to inclusion pathology in ALS, FTLD-TDP and AD. PMID:22993125

  1. Body size change in various nematodes depending on bacterial food, sex and growth temperature.

    PubMed

    So, Shuhei; Garan, Yohei; Miyahara, Kohji; Ohshima, Yasumi

    2012-04-01

    We previously reported significant body size change in the nematode Caenorhabditis elegans, depending on the food strain of E. coli. Here, we examined this body size change in 11 other nematode species as well, and found that it is common to most of these nematodes. Furthermore, this food-dependent body size change is influenced by sex and growth temperature. PMID:24058830

  2. Body size change in various nematodes depending on bacterial food, sex and growth temperature

    PubMed Central

    So, Shuhei; Garan, Yohei; Miyahara, Kohji; Ohshima, Yasumi

    2012-01-01

    We previously reported significant body size change in the nematode Caenorhabditis elegans, depending on the food strain of E. coli. Here, we examined this body size change in 11 other nematode species as well, and found that it is common to most of these nematodes. Furthermore, this food-dependent body size change is influenced by sex and growth temperature. PMID:24058830

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

    USGS Publications Warehouse

    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.

  4. Molecular events linking cholesterol to Alzheimer’s disease and inclusion body myositis in a rabbit model

    PubMed Central

    Liu, Qing Yan; Koukiekolo, Roger; Zhang, Dong Ling; Smith, Brandon; Ly, Dao; Lei, Joy X; Ghribi, Othman

    2016-01-01

    Alzheimer’s disease (AD) is the most common neurodegenerative disorder, characterized by cognitive impairment and dementia, resulting from progressive synaptic dysfunction, loss and neuronal cell death. Inclusion body myositis (IBM) is a skeletal muscle degenerative disease, displaying progressive proximal and distal muscle weakness, in association with muscle fiber atrophy, degeneration and death. Studies have shown that the late onset version of AD (LOAD) and sporadic IBM (sIBM) in muscle share many pathological features, including the presence of extracellular plaques of β-amyloid peptides and intracellular tangles of hyperphosphorylated tau proteins. High blood cholesterol is suggested to be a risk factor for LOAD. Many neuropathological changes of LOAD can be reproduced by feeding rabbits a 2% enriched cholesterol diet for 12 weeks. The cholesterol fed rabbit model also simultaneously develops sIBM like pathology, which makes it an ideal model to study the molecular mechanisms common to the development of both diseases. In the present study, we determined the changes of gene expression in rabbit brain and muscle during the progression of LOAD and sIBM pathology using a custom rabbit nucleotide microarray, followed by qRT-PCR analyses. Out of 869 unique transcripts screened, 47 genes showed differential expression between the control and the cholesterol-treated group during the 12 week period and 19 changed transcripts appeared to be common to LOAD and sIBM. The most notable changes are the upregulation of the hemoglobin gene family and the downregulation of the genes required for mitochondrial oxidative phosphorylation in both brain and muscle tissues throughout the time course. The significant overlap on the changes of gene expression in the brain and muscle of rabbits fed with cholesterol-enriched diet supports the notion that LOAD and sIBM may share a common etiology. PMID:27073745

  5. Light chain of botulinum A neurotoxin expressed as an inclusion body from a synthetic gene is catalytically and functionally active.

    PubMed

    Ahmed, S A; Smith, L A

    2000-08-01

    Botulinum neurotoxins, the most potent of all toxins, induce lethal neuromuscular paralysis by inhibiting exocytosis at the neuromuscular junction. The light chains (LC) of these dichain neurotoxins are a new class of zinc-endopeptidases that specifically cleave the synaptosomal proteins, SNAP-25, VAMP, or syntaxin at discrete sites. To facilitate the structural and functional characterization of these unique endopeptidases, we constructed a synthetic gene for the LC of the botulinum neurotoxin serotype A (BoNT/A), overexpressed it in Escherichia coli, and purified the gene product from inclusion bodies. Our procedure can provide 1.1 g of the LC from 1 L of culture. The LC product was stable in solution at 4 degrees C for at least 6 months. This rBoNT/A LC was proteolytically active, specifically cleaving the Glu-Arg bond in a 17-residue synthetic peptide of SNAP-25, the reported cleavage site of BoNT/A. Its calculated catalytic efficiency kcat/Km was higher than that reported for the native BoNT/A dichain. Treating the rBoNT/A LC with mercuric compounds completely abolished its activity, most probably by modifying the cysteine-164 residue located in the vicinity of the active site. About 70% activity of the LC was restored by adding Zn2+ to a Zn2+-free, apo-LC preparation. The LC was nontoxic to mice and failed to elicit neutralizing epitope(s) when the animals were vaccinated with this protein. In addition, injecting rBoNT/A LC into sea urchin eggs inhibited exocytosis-dependent plasma membrane resealing. For the first time, results of our study make available a large amount of the biologically active toxin fragment in a soluble and stable form. PMID:11195972

  6. A retrospective cohort study identifying the principal pathological features useful in the diagnosis of inclusion body myositis

    PubMed Central

    Brady, Stefen; Squier, Waney; Sewry, Caroline; Hanna, Michael; Hilton-Jones, David; Holton, Janice L

    2014-01-01

    Objective The current pathological diagnostic criteria for sporadic inclusion body myositis (IBM) lack sensitivity. Using immunohistochemical techniques abnormal protein aggregates have been identified in IBM, including some associated with neurodegenerative disorders. Our objective was to investigate the diagnostic utility of a number of markers of protein aggregates together with mitochondrial and inflammatory changes in IBM. Design Retrospective cohort study. The sensitivity of pathological features was evaluated in cases of Griggs definite IBM. The diagnostic potential of the most reliable features was then assessed in clinically typical IBM with rimmed vacuoles (n=15), clinically typical IBM without rimmed vacuoles (n=9) and IBM mimics—protein accumulation myopathies containing rimmed vacuoles (n=7) and steroid-responsive inflammatory myopathies (n=11). Setting Specialist muscle services at the John Radcliffe Hospital, Oxford and the National Hospital for Neurology and Neurosurgery, London. Results Individual pathological features, in isolation, lacked sensitivity and specificity. However, the morphology and distribution of p62 aggregates in IBM were characteristic and in a myopathy with rimmed vacuoles, the combination of characteristic p62 aggregates and increased sarcolemmal and internal major histocompatibility complex class I expression or endomysial T cells were diagnostic for IBM with a sensitivity of 93% and specificity of 100%. In an inflammatory myopathy lacking rimmed vacuoles, the presence of mitochondrial changes was 100% sensitive and 73% specific for IBM; characteristic p62 aggregates were specific (91%), but lacked sensitivity (44%). Conclusions We propose an easily applied diagnostic algorithm for the pathological diagnosis of IBM. Additionally our findings support the hypothesis that many of the pathological features considered typical of IBM develop later in the disease, explaining their poor sensitivity at disease presentation and

  7. Production of soluble truncated spike protein of porcine epidemic diarrhea virus from inclusion bodies of Escherichia coli through refolding.

    PubMed

    Piao, Da-Chuan; Lee, Yoon-Seok; Bok, Jin-Duck; Cho, Chong-Su; Hong, Zhong-Shan; Kang, Sang-Kee; Choi, Yun-Jaie

    2016-10-01

    The emergence of highly pathogenic variant porcine epidemic diarrhea virus (PEDV) strains, from 2013 to 2014, in North American and Asian countries have greatly threatened global swine industry. Therefore, development of effective vaccines against PEDV variant strains is urgently needed. Recently, it has been reported that the N-terminal domain (NTD) of S1 domain of PEDV spike protein is responsible for binding to the 5-N-acetylneuraminic acid (Neu5Ac), a possible sugar co-receptor. Therefore, the NTD of S1 domain could be an attractive target for the development of subunit vaccines. In this study, the NTD spanning amino acid residues 25-229 (S25-229) of S1 domain of PEDV variant strain was expressed in Escherichia coli BL21 (DE3) in the form of inclusion bodies (IBs). S25-229 IBs were solubilized in 20 mM sodium acetate (pH 4.5) buffer containing 8 M urea and 1 mM dithiothreitol with 95% yield. Solubilized S25-229 IBs were refolded by 10-fold flash dilution and purified by one-step cation exchange chromatography with >95% purity and 20% yield. The CD spectrum of S25-229 showed the characteristic pattern of alpha helical structure. In an indirect ELISA, purified S25-229 showed strong reactivity with mouse anti-PEDV sera. In addition, immunization of mice with 20 μg of purified S25-229 elicited highly potent serum IgG titers. Finally, mouse antisera against S25-229 showed immune reactivity with native PEDV S protein in an immunofluorescence assay. These results suggest that purified S25-229 may have potential to be used as a subunit vaccine against PEDV variant strains. PMID:27260969

  8. Recombinant production of biologically active giant grouper (Epinephelus lanceolatus) growth hormone from inclusion bodies of Escherichia coli by fed-batch culture.

    PubMed

    Chung, Wen-Jen; Huang, Chi-Lung; Gong, Hong-Yi; Ou, Tsung-Yin; Hsu, Jue-Liang; Hu, Shao-Yang

    2015-06-01

    Growth hormone (GH) performs important roles in regulating somatic growth, reproduction, osmoregulation, metabolism and immunity in teleosts, and thus, it has attracted substantial attention in the field of aquaculture application. Herein, giant grouper GH (ggGH) cDNA was cloned into the pET28a vector and expressed in Shuffle® T7 Competent Escherichia coli. Recombinant N-terminal 6× His-tagged ggGH was produced mainly in insoluble inclusion bodies; the recombinant ggGH content reached 20% of total protein. For large-scale ggGH production, high-cell density E. coli culture was achieved via fed-batch culture with pH-stat. After 30h of cultivation, a cell concentration of 41.1g/l dry cell weight with over 95% plasmid stability was reached. Maximal ggGH production (4.0g/l; 22% total protein) was achieved via mid-log phase induction. Various centrifugal forces, buffer pHs and urea concentrations were optimized for isolation and solubilization of ggGH from inclusion bodies. Hydrophobic interactions and ionic interactions were the major forces in ggGH inclusion body formation. Complete ggGH inclusion body solubilization was obtained in PBS buffer at pH 12 containing 3M urea. Through a simple purification process including Ni-NTA affinity chromatography and refolding, 5.7mg of ggGH was obtained from 10ml of fed-batch culture (45% recovery). The sequence and secondary structure of the purified ggGH were confirmed by LC-MS/MS mass spectrometry and circular dichroism analysis. The cell proliferation-promoting activity was confirmed in HepG2, ZFL and GF-1 cells with the WST-1 colorimetric bioassay. PMID:25703054

  9. Structural studies on the polyhedral inclusion bodies, virions, and DNA of the nuclear polyhedrosis virus of the cotton bollworm Heliothis zea.

    PubMed Central

    Scharnhorst, D W; Saving, K L; Vuturo, S B; Cooke, P H; Weaver, R F

    1977-01-01

    The polyhedral inclusion body of the cotton bollworm nuclear polyhedrosis virus contains virions occluded in an orthogonal crystalline matrix. The virions appear as rods or, more frequently, as oval structures that form upon bending of the nucleocapsid within the viral membrane. The nucleocapsid consists at least of DNA surrounded by a capsid composed of subunits, possibly helically arranged. The viral DNA is circular and supercoiled. It is heterogenous in size with contour lengths ranging from 15 to 45 mum. Images PMID:319251

  10. L-cysteine-enhanced renaturation of bioactive soluble tumor necrosis factor ligand family member LIGHT from inclusion bodies in Escherichia coli.

    PubMed

    Tsuji, Isamu; Mastui, Hideki; Ito, Tatsuo; Kurokawa, Tomofumi; Shintani, Yasushi

    2011-12-01

    LIGHT is a membrane-bound protein that belongs to the tumor necrosis factor (TNF) superfamily ligands. In this study, we established an effective strategy for producing a bioactive soluble form of LIGHT (sLIGHT), an extracellular region (Ile⁸⁴-Val²⁴⁰) of human LIGHT. Because sLIGHT was expressed as inclusion bodies in Escherichia coli, we investigated reagents that enhance the renaturation of sLIGHT from the inclusion bodies. Interestingly, L-cysteine in the denaturation buffer containing 3.5 M guanidine hydrochloride significantly improved the renaturation efficiency of sLIGHT. The effect of L-cysteine was synergistically enhanced by L-arginine in the refolding buffer. The optimal concentrations of L-cysteine and L-arginine in the denaturation and refolding buffers were 8 mM and 0.8 M, respectively. With these buffers, approximately 90 mg of sLIGHT was purified from 200 g of frozen E. coli cells. sLIGHT thus obtained significantly induced apoptosis in the WiDr human colon adenocarcinoma cell line at nanomolar concentrations, the same amount of sLIGHT that was produced by Sf9 insect cells. These results suggest that L-cysteine in the denaturation buffer enhances the renaturation of recombinant proteins from inclusion bodies in E. coli. PMID:21745574

  11. VCP Associated Inclusion Body Myopathy and Paget Disease of Bone Knock-In Mouse Model Exhibits Tissue Pathology Typical of Human Disease

    PubMed Central

    Kitazawa, Masashi; Su, Hailing; Tanaja, Jasmin; Dec, Eric; Wallace, Douglas C.; Mukherjee, Jogeshwar; Caiozzo, Vincent; Warman, Matthew; Kimonis, Virginia E.

    2010-01-01

    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 VCPR155H/+ knock-in mice replicate the muscle, bone and brain pathology of inclusion body myopathy, thus representing a useful model for preclinical studies. PMID:20957154

  12. Bacterial antigen detection in body fluids: methods for rapid antigen concentration and reduction of nonspecific reactions.

    PubMed Central

    Doskeland, S O; Berdal, B P

    1980-01-01

    We sought procedures which would allow a rapid concentration in high yield of bacterial antigens from tissue fluids of patients and which could be applied also to protein-rich fluids like serum. Ethanol precipitation at a subzero temperature with albumin added as an antigen coprecipitant made it possible to achieve a more than 20-fold concentration of antigen in 15 min and a 200-fold concentration in 45 min. Heat-stable antigens could be concentrated from protein-rich fluids (like serum) after the sample had been deproteinized by boiling. Such heating (100 degrees C, 3 min) also liberated bacterial polysaccharides from antibody complexes and elminated the nonspecific interference of serum in enzyme-linked immunosorbent assay. PMID:7372801

  13. Mineralogy and Petrology of Amoeboid Olivine Inclusions in CO3 Chondrites: Relationship to Parent-Body Aqueous Alteration

    NASA Technical Reports Server (NTRS)

    Chizmadia, Lysa J.; Rubin, Alan E.; Wasson, John T.

    2003-01-01

    Petrographic and mineralogic studies of amoeboid olivine inclusions (AOIs) in CO3 carbonaceous chondrites reveal that they are sensitive indicators of parent-body aqueous and thermal alteration. As the petrologic subtype increases from 3.0 to 3.8, forsteritic olivine (Fa(sub 0-1)) is systematically converted into ferroan olivine (Fa(sub 60-75)). We infer that the Fe, Si and O entered the assemblage along grain boundaries, forming ferroan olivine that filled fractures and voids. As temperatures increased, Fe(+2) from the new olivine exchanged with Mg(+2) from the original AOI to form diffusive haloes around low-FeO cores. Cations of Mn(+2), Ca(+2) and Cr(+3) were also mobilized. The systematic changes in AOI textures and olivine compositional distributions can be used to refine the classification of CO3 chondrites into subtypes. In subtype 3.0, olivine occurs as small forsterite grains (Fa(sub 0-1)), free of ferroan olivine. In petrologic subtype 3.2, narrow veins of FeO-rich olivine have formed at forsterite grain boundaries. With increasing alteration, these veins thicken to form zones of ferroan olivine at the outside AOI margin and within the AOI interior. By subtype 3.7, there is a fairly broad olivine compositional distribution in the range Fa(sub 63-70), and by subtype 3.8, no forsterite remains and the high-Fa peak has narrowed, Fa(sub 64-67). Even at this stage, there is incomplete equilibration in the chondrite as a whole (e.g., data for coarse olivine grains in Isna (CO3.8) chondrules and lithic clasts show a peak at Fa(sub39)). We infer that the mineral changes in A01 identified in the low petrologic types required aqueous or hydrothermal fluids whereas those in subtypes greater than or equal to 3.3 largely reflect diffusive exchange within and between mineral grains without the aid of fluids.

  14. Blocking monocyte transmigration in in vitro system by a human antibody scFv anti-CD99. Efficient large scale purification from periplasmic inclusion bodies in E. coli expression system.

    PubMed

    Moricoli, Diego; Muller, William Anthony; Carbonella, Damiano Cosimo; Balducci, Maria Cristina; Dominici, Sabrina; Watson, Richard; Fiori, Valentina; Weber, Evan; Cianfriglia, Maurizio; Scotlandi, Katia; Magnani, Mauro

    2014-06-01

    Migration of leukocytes into site of inflammation involves several steps mediated by various families of adhesion molecules. CD99 play a significant role in transendothelial migration (TEM) of leukocytes. Inhibition of TEM by specific monoclonal antibody (mAb) can provide a potent therapeutic approach to treating inflammatory conditions. However, the therapeutic utilization of whole IgG can lead to an inappropriate activation of Fc receptor-expressing cells, inducing serious adverse side effects due to cytokine release. In this regard, specific recombinant antibody in single chain variable fragments (scFvs) originated by phage library may offer a solution by affecting TEM function in a safe clinical context. However, this consideration requires large scale production of functional scFv antibodies and the absence of toxic reagents utilized for solubilization and refolding step of inclusion bodies that may discourage industrial application of these antibody fragments. In order to apply the scFv anti-CD99 named C7A in a clinical setting, we herein describe an efficient and large scale production of the antibody fragments expressed in E. coli as periplasmic insoluble protein avoiding gel filtration chromatography approach, and laborious refolding step pre- and post-purification. Using differential salt elution which is a simple, reproducible and effective procedure we are able to separate scFv in monomer format from aggregates. The purified scFv antibody C7A exhibits inhibitory activity comparable to an antagonistic conventional mAb, thus providing an excellent agent for blocking CD99 signaling. This protocol can be useful for the successful purification of other monomeric scFvs which are expressed as periplasmic inclusion bodies in bacterial systems. PMID:24798881

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

    Cancer.gov

    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

  16. Modeling the bacterial flagellum by an elastic network of rigid bodies

    NASA Astrophysics Data System (ADS)

    Speier, C.; Vogel, R.; Stark, H.

    2011-08-01

    Bacteria such as Escherichia coli propel themselves by rotating a bundle of helical filaments, each driven by a rotary motor embedded in the cell membrane. Each filament is an assembly of thousands of copies of the protein flagellin which assumes two different states. We model the filament by an elastic network of rigid bodies that form bonds with one another according to a scheme suggested by Namba and Vondervistz (1997 Q. Rev. Biophys. 30 1-65) and add additional binding sites at the inner part of the rigid body. Our model reproduces the helical parameters of the 12 possible polymorphic configurations very well. We demonstrate that its energetical ground state corresponds to the normal helical form, usually observed in nature, only when inner and outer binding sites of the rigid body have a large axial displacement. This finding correlates directly to the elongated shape of the flagellin molecule. An Ising Hamiltonian in our model directly addresses the two states of the flagellin protein. It contains an external field that represents external parameters which allow us to alter the ground state of the filament.

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

    PubMed Central

    White, Matthew D.; Bosio, Catharine M.; Duplantis, Barry N.

    2012-01-01

    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. PMID:21626408

  18. Importin-α7 Is Involved in the Formation of Ebola Virus Inclusion Bodies but Is Not Essential for Pathogenicity in Mice.

    PubMed

    Gabriel, Gülsah; Feldmann, Friederike; Reimer, Rudolph; Thiele, Swantje; Fischer, Meike; Hartmann, Enno; Bader, Michael; Ebihara, Hideki; Hoenen, Thomas; Feldmann, Heinz

    2015-10-01

    Ebola virus (EBOV) protein 24 antagonizes the host interferon (IFN) response by hijacking select nuclear importin-α isoforms. Thereby, it blocks STAT1-mediated IFN-α/β and IFN-γ synthesis. However, owing to the lack of importin-α knockout animal models in the past, their role in EBOV pathogenesis remained largely unknown. Here, we demonstrate that importin-α7 is involved in the formation of EBOV inclusion bodies and replication. However, deletion of the gene encoding importin-α7 was not sufficient to increase survival rates among mice infected with EBOV. PMID:26185094

  19. Identification, Characterization, and In Vitro Culture of Highly Divergent Arenaviruses from Boa Constrictors and Annulated Tree Boas: Candidate Etiological Agents for Snake Inclusion Body Disease

    PubMed Central

    Stenglein, Mark D.; Sanders, Chris; Kistler, Amy L.; Ruby, J. Graham; Franco, Jessica Y.; Reavill, Drury R.; Dunker, Freeland; DeRisi, Joseph L.

    2012-01-01

    ABSTRACT Inclusion body disease (IBD) is an infectious fatal disease of snakes typified by behavioral abnormalities, wasting, and secondary infections. At a histopathological level, the disease is identified by the presence of large eosinophilic cytoplasmic inclusions in multiple tissues. To date, no virus or other pathogen has been definitively characterized or associated with the disease. Using a metagenomic approach to search for candidate etiologic agents in snakes with confirmed IBD, we identified and de novo assembled the complete genomic sequences of two viruses related to arenaviruses, and a third arenavirus-like sequence was discovered by screening an additional set of samples. A continuous boa constrictor cell line was established and used to propagate and isolate one of the viruses in culture. Viral nucleoprotein was localized and concentrated within large cytoplasmic inclusions in infected cells in culture and tissues from diseased snakes. In total, viral RNA was detected in 6/8 confirmed IBD cases and 0/18 controls. These viruses have a typical arenavirus genome organization but are highly divergent, belonging to a lineage separate from that of the Old and New World arenaviruses. Furthermore, these viruses encode envelope glycoproteins that are more similar to those of filoviruses than to those of other arenaviruses. These findings implicate these viruses as candidate etiologic agents of IBD. The presence of arenaviruses outside mammals reveals that these viruses infect an unexpectedly broad range of species and represent a new reservoir of potential human pathogens. PMID:22893382

  20. On the effect of inertia and history forces on the slow motion of a spherical solid or gaseous inclusion in a solid-body rotation flow

    NASA Astrophysics Data System (ADS)

    Candelier, Fabien; Angilella, Jean-Régis; Souhar, Mohamed

    2005-12-01

    The motion of a spherical inclusion released in a vertical solid-body rotation flow is investigated theoretically and experimentally. Solid spheres and bubbles are considered. The particle Reynolds number, the Taylor number, the Weber number and the capillary number are smaller than unity. The motion equations of the inclusion are obtained by revisiting the hydrodynamic equations. The axial (vertical) motion and the horizontal motion are uncoupled, even though they are sensitive to the rotation rate of the flow. Analytical solutions of the particle motion equation are compared to experimental results obtained by releasing a particle in a rotating tank filled with silicone oil. For solid spheres and bubbles, both the terminal velocity and the particle ejection rate (or trapping rate) predicted by the theory agree with experiments, without any empirical adjustment. In particular, the experimental device enables us to check the validity of various theories involving solid or gaseous inclusions with or without inertia or history effects. It is observed that the mobility tensor obtained by writing the fluid motion equations in the rotating frame accurately predicts the horizontal particle trajectory, like the Boussinesq-Basset equation obtained by writing the fluid motion equations in the non-rotating frame and neglecting the horizontal contribution of inertia effects.

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

    PubMed

    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. PMID:23688859

  2. Computational biomechanics of human brain with and without the inclusion of the body under different blast orientation.

    PubMed

    Salimi Jazi, Mehdi; Rezaei, Asghar; Azarmi, Fardad; Ziejewski, Mariusz; Karami, Ghodrat

    2016-07-01

    Three different human head models in a free space are exposed to blast waves coming from four different directions. The four head-neck-body models composed of model a, with the neck free in space; model b, with neck fixed at the bottom; and model c, with the neck attached to the body. The results show that the effect of the body can be ignored for the first milliseconds of the head-blast wave interactions. Also one can see that although most biomechanical responses of the brain have similar patterns in all models, the shear stresses are heavily increased after a few milliseconds in model b in which the head motion is obstructed by the fixed-neck boundary conditions. The free-floating head model results are closer to the attached-body model. PMID:26442577

  3. Establishment of Mouse Model of MYH9 Disorders: Heterozygous R702C Mutation Provokes Macrothrombocytopenia with Leukocyte Inclusion Bodies, Renal Glomerulosclerosis and Hearing Disability

    PubMed Central

    Suzuki, Nobuaki; Kunishima, Shinji; Ikejiri, Makoto; Maruyama, Shoichi; Sone, Michihiko; Takagi, Akira; Ikawa, Masahito; Okabe, Masaru; Kojima, Tetsuhito; Saito, Hidehiko; Naoe, Tomoki; Matsushita, Tadashi

    2013-01-01

    Nonmuscle myosin heavy chain IIA (NMMHCIIA) encoded by MYH9 is associated with autosomal dominantly inherited diseases called MYH9 disorders. MYH9 disorders are characterized by macrothrombocytopenia and very characteristic inclusion bodies in granulocytes. MYH9 disorders frequently cause nephritis, sensorineural hearing disability and cataracts. One of the most common and deleterious mutations causing these disorders is the R702C missense mutation. We generated knock-in mice expressing the Myh9 R702C mutation. R702C knock-in hetero mice (R702C+/− mice) showed macrothrombocytopenia. We studied megakaryopoiesis of cultured fetal liver cells of R702C+/− mice and found that proplatelet formation was impaired: the number of proplatelet tips was decreased, proplatelet size was increased, and proplatelet shafts were short and enlarged. Although granulocyte inclusion bodies were not visible by May–Grünwald Giemsa staining, immunofluorescence analysis indicated that NMMHCIIA proteins aggregated and accumulated in the granulocyte cytoplasm. In other organs, R702C+/− mice displayed albuminuria which increased with age. Renal pathology examination revealed glomerulosclerosis. Sensory hearing loss was indicated by lowered auditory brainstem response. These findings indicate that Myh9 R702C knock-in mice mirror features of human MYH9 disorders arising from the R702C mutation. PMID:23976996

  4. Quantitative relationships between huntingtin levels, polyglutamine length, inclusion body formation, and neuronal death provide novel insight into Huntington’s disease molecular pathogenesis

    PubMed Central

    Miller, Jason; Arrasate, Montserrat; Shaby, Benjamin A.; Mitra, Siddhartha; Masliah, Eliezer; Finkbeiner, Steven

    2010-01-01

    An expanded polyglutamine (polyQ) stretch in the protein huntingtin (htt) induces self-aggregation into inclusion bodies (IBs) and causes Huntington’s disease (HD). Defining precise relationships between early observable variables and neuronal death at the molecular and cellular levels should improve our understanding of HD pathogenesis. Here, we utilized an automated microscope that can track thousands of neurons individually over their entire lifetime to quantify interconnected relationships between early variables, such as htt levels, polyQ length, and IB formation, and neuronal death in a primary striatal model of HD. The resulting model revealed that: mutant htt increases the risk of death by tonically interfering with homeostatic coping mechanisms rather than producing accumulated damage to the neuron; htt toxicity is saturable; the rate limiting steps for inclusion body formation and death can be traced to different conformational changes in monomeric htt; and IB formation reduces the impact of a neuron’s starting levels of htt on its risk of death. Finally, the model that emerges from our quantitative measurements places critical limits on the potential mechanisms by which mutant htt might induce neurodegeneration, which should help direct future research. PMID:20685997

  5. The inclusion of drag forces in the restricted problem of three bodies and the instability of the triangular points.

    NASA Astrophysics Data System (ADS)

    Danby, J. M. A.

    If the restricted problem of three bodies is modified so that the third body experiences a drag force proportional to its velocity relative to a surrounding medium which is modelled so that the triangular points remain equilibria, then, in a linear analysis these points are unstable. The long-period oscillations become associated with an exponential factor eat with positive a. This has been confirmed by numerical integration of the complete nonlinear equations. Variations in the kinematics of the surrounding medium and in its density are also considered: the instability remains.

  6. The Inclusive Classroom: How Inclusive Is Inclusion?

    ERIC Educational Resources Information Center

    Reid, Claudette M.

    2010-01-01

    This paper presents the position that inclusion is limited; inclusion does not go far enough. The inclusive classroom has been assessed to be of benefit both to the teacher and student. There are, however, limits set on inclusion. In most classrooms only children with learning disability are included omitting those with severe disabilities,…

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

    PubMed Central

    Tokuda, Gaku; Lo, Nathan; Takase, Aya; Yamada, Akinori; Hayashi, Yoshinobu; Watanabe, Hirofumi

    2008-01-01

    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. PMID:19025664

  8. Real-time monitoring of bacterial infection in vivo: development of bioluminescent staphylococcal foreign-body and deep-thigh-wound mouse infection models.

    PubMed

    Kuklin, Nelly A; Pancari, Gregory D; Tobery, Timothy W; Cope, Leslie; Jackson, Jesse; Gill, Charles; Overbye, Karen; Francis, Kevin P; Yu, Jun; Montgomery, Donna; Anderson, Annaliesa S; McClements, William; Jansen, Kathrin U

    2003-09-01

    Staphylococcal infections associated with catheter and prosthetic implants are difficult to eradicate and often lead to chronic infections. Development of novel antibacterial therapies requires simple, reliable, and relevant models for infection. Using bioluminescent Staphylococcus aureus, we have adapted the existing foreign-body and deep-wound mouse models of staphylococcal infection to allow real-time monitoring of the bacterial colonization of catheters or tissues. This approach also enables kinetic measurements of bacterial growth and clearance in each infected animal. Persistence of infection was observed throughout the course of the study until termination of the experiment at day 16 in a deep-wound model and day 21 in the foreign-body model, providing sufficient time to test the effects of antibacterial compounds. The usefulness of both animal models was assessed by using linezolid as a test compound and comparing bioluminescent measurements to bacterial counts. In the foreign-body model, a three-dose antibiotic regimen (2, 5, and 24 h after infection) resulted in a decrease in both luminescence and bacterial counts recovered from the implant compared to those of the mock-treated infected mice. In addition, linezolid treatment prevented the formation of subcutaneous abscesses, although it did not completely resolve the infection. In the thigh model, the same treatment regimen resulted in complete resolution of the luminescent signal, which correlated with clearance of the bacteria from the thighs. PMID:12936968

  9. Explicit finite element modelling of heel pad mechanics in running: inclusion of body dynamics and application of physiological impact loads.

    PubMed

    Chen, Wen-Ming; Lee, Peter Vee-Sin

    2015-01-01

    Many heel pathologies including plantar heel pain may result from micro tears/trauma in the subcutaneous tissues, in which internal tissue deformation/stresses within the heel pad play an important role. Previously, many finite element models have been proposed to evaluate stresses inside the heel pad, but the majority of these models only focus on static loading boundary conditions. This study explored a dynamics modelling approach to the heel pad subjected to realistic impact loads during running. In this model, the inertial property and action of the body are described by a lumped parameter model, while the heel/shoe interactions are modelled using a viscoelastic heel pad model with contact properties. The impact force pattern, dynamic heel pad deformation and stress states predicted by the model were compared with published experimental data. Further parametrical studies revealed the model responses, in terms of internal stresses in the skin and fatty tissue, change nonlinearly when body dynamics changes. A reduction in foot's touchdown velocity resulted in a less severe impact landing and stress relief inside the heel pad, for example peak von-Mises stress in fatty tissue, was reduced by 11.3%. Applications of the model may be extendable to perform iterative analyses to further understand the complex relationships between body dynamics and stress distributions in the soft tissue of heel pad during running. This may open new opportunities to study the mechanical aetiology of plantar heel pain in runners. PMID:24980181

  10. Silica-coated magnetic nanoparticles impair proteasome activity and increase the formation of cytoplasmic inclusion bodies in vitro

    PubMed Central

    Phukan, Geetika; Shin, Tae Hwan; Shim, Jeom Soon; Paik, Man Jeong; Lee, Jin-Kyu; Choi, Sangdun; Kim, Yong Man; Kang, Seong Ho; Kim, Hyung Sik; Kang, Yup; Lee, Soo Hwan; Mouradian, M. Maral; Lee, Gwang

    2016-01-01

    The potential toxicity of nanoparticles, particularly to neurons, is a major concern. In this study, we assessed the cytotoxicity of silica-coated magnetic nanoparticles containing rhodamine B isothiocyanate dye (MNPs@SiO2(RITC)) in HEK293 cells, SH-SY5Y cells, and rat primary cortical and dopaminergic neurons. In cells treated with 1.0 μg/μl MNPs@SiO2(RITC), the expression of several genes related to the proteasome pathway was altered, and proteasome activity was significantly reduced, compared with control and with 0.1 μg/μl MNPs@SiO2(RITC)-treated cells. Due to the reduction of proteasome activity, formation of cytoplasmic inclusions increased significantly in HEK293 cells over-expressing the α–synuclein interacting protein synphilin-1 as well as in primary cortical and dopaminergic neurons. Primary neurons, particularly dopaminergic neurons, were more vulnerable to MNPs@SiO2(RITC) than SH-SY5Y cells. Cellular polyamines, which are associated with protein aggregation, were significantly altered in SH-SY5Y cells treated with MNPs@SiO2(RITC). These findings highlight the mechanisms of neurotoxicity incurred by nanoparticles. PMID:27378605

  11. Chorea as a clinical feature of the basophilic inclusion body disease subtype of fused-in-sarcoma-associated frontotemporal lobar degeneration.

    PubMed

    Kawakami, Ito; Kobayashi, Zen; Arai, Tetsuaki; Yokota, Osamu; Nonaka, Takashi; Aoki, Naoya; Niizato, Kazuhiro; Oshima, Kenichi; Higashi, Shinji; Katsuse, Omi; Hosokawa, Masato; Hasegawa, Masato; Akiyama, Haruhiko

    2016-01-01

    Choreoathetoid involuntary movements are rarely reported in patients with frontotemporal lobar degeneration (FTLD), suggesting their exclusion as a supportive feature in clinical diagnostic criteria for FTLD. Here, we identified three cases of the behavioral variant of frontotemporal dementia (bvFTD) that display chorea with fused in sarcoma (FUS)-positive inclusions (FTLD-FUS) and the basophilic inclusion body disease (BIBD) subtype. We determined the behavioral and cognitive features in this group that were distinct from other FTLD-FUS cases. We also reviewed the clinical records of 72 FTLD cases, and clarified additional clinical features that are predictive of the BIBD pathology. Symptom onset in the three patients with chorea was at 44.0 years of age (±12.0 years), and occurred in the absence of a family history of dementia. The cases were consistent with a clinical form of FTD known as bvFTD, as well as reduced neurological muscle tone in addition to chorea. The three patients showed no or mild parkinsonism, which by contrast, increased substantially in the other FTLD cases until a later stage of disease. The three patients exhibited severe caudate atrophy, which has previously been reported as a histological feature distinguishing FTLD-FUS from FTLD-tau or FTLD-TAR DNA-binding protein 43. Thus, our findings suggest that the clinical feature of choreoathetosis in bvFTD might be associated with FTLD-FUS, and in particular, with the BIBD subtype. PMID:27044537

  12. Measles Inclusion-Body Encephalitis: Neuronal Phosphorylated Tau Protein is Present in the Biopsy but not in the Autoptic Specimens of the Same Patient.

    PubMed

    Maderna, Emanuela; Fugnanesi, Valeria; Morbin, Michela; Cacciatore, Francesca; Spinello, Sonia; Godani, Massimiliano; Zoia, Riccardo; Tagliavini, Fabrizio; Giaccone, Giorgio

    2016-07-01

    Tauopathies are sporadic or familial neurodegenerative diseases characterized by the accumulation of phosphorylated tau in neurons and glial cells and include encephalitis related to measles virus such as subacute sclerosing panencephalitis. We describe a 45-year-old woman, with a history of lymphoma treated with immunosuppressant therapy who underwent an open biopsy of the right frontal cortex for a suspect of encephalitis, and died 4 days later. The neuropathological assessment on the bioptic sample revealed edema, severe gliosis and microglial activation, with lymphomonocytic perivascular cuffing and neurons containing both nuclear and cytoplasmic eosinofilic inclusions that ultrastructurally appeared as tubular and curvilinear non-membrane-bound 12-18 nm structures, leading to the diagnosis of measles inclusion-bodies encephalitis. The biopsy specimen showed several cortical neurons with intense perikaryal immunoreactivity for anti-tau antibodies recognizing phosphorylated epitopes while on autoptic specimens no phosphorylated tau immunoreactivity was detected. Our findings suggest that in specific conditions biopsy-derived human tau may be phosphorylated at sites that may result not phosphorylated in autopsy-derived specimens, most likely caused by post-mortem dephosphorylation. PMID:26462994

  13. Muscle weakness correlates with muscle atrophy and precedes the development of inclusion body or rimmed vacuoles in the mouse model of DMRV/hIBM.

    PubMed

    Malicdan, May Christine V; Noguchi, Satoru; Hayashi, Yukiko K; Nishino, Ichizo

    2008-09-17

    Distal myopathy with rimmed vacuoles (DMRV), also called hereditary inclusion body myopathy (hIBM), is characterized clinically by weakness and atrophy that initially involves the distal muscles and pathologically by the presence of rimmed vacuoles (RVs) or intracellular protein deposits in myofibers. It is caused by mutations in the UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) gene that is important in sialic acid synthesis. Recently, we generated a mouse model (Gne(-/-)hGNED176VTg) that exhibits muscle weakness and pathological changes similar to DMRV patients. To gain better understanding of the pathomechanism of DMRV, we determined temporal changes in the overall motor performance of this model mouse for DMRV in correlation with the structure and function of isolated skeletal muscles and muscle pathology. These DMRV mice exhibited muscle weakness, decreased whole muscle mass and cross-sectional area (CSA), and reduced contractile power in an age-related manner. Single-fiber CSA further supported the finding of muscle atrophy that involved both type I and type II fibers. These results suggest that atrophy is highly correlated with reduced production of force at young age, both in vivo and ex vivo, thereby implicating the important role of atrophy in the pathomechanism of DMRV. In older age, and particularly in gastrocnemius muscles, RVs and intracellular inclusions were seen in type IIA fibers, further aggravating reduction of force and specific increase in twitch-tetanus ratio. PMID:18628337

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

    PubMed

    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. PMID:24647109

  15. Penile epidermal inclusion cyst.

    PubMed

    Saini, Pradeep; Mansoor, M N; Jalali, Sanjay; Sharma, Abhishek

    2010-07-01

    We report a case of epidermal inclusion cyst of penis in a five-year-old boy, who had presented to the outpatient department of our hospital. Epidermal inclusion cysts are benign lesions that can develop in any part of the body. However, the finding of an epidermal inclusion cyst in the penis is rare. The child was operated and discharged uneventfully. The objective of reporting this case is to highlight the rare possibility of an inclusion cyst arising from penis as a late complication of circumcision. PMID:20589475

  16. 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.

    PubMed

    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. PMID:26782747

  17. Pathogenicity and Complete Genome Characterization of Fowl Adenoviruses Isolated from Chickens Associated with Inclusion Body Hepatitis and Hydropericardium Syndrome in China

    PubMed Central

    Zhao, Jing; Zhong, Qi; Zhao, Ye; Hu, Yan-xin; Zhang, Guo-zhong

    2015-01-01

    In this study, we determined and genetically characterized three fowl adenoviruses isolated from chickens with inclusion body hepatitis (IBH) and hydropericardium syndrome (HPS) in China and assessed their pathogenicity. The full genome of HBQ12, BJH13 and JSJ13 was found to be 44,081, 43,966 and 43,756 nucleotides long, respectively. Sequence alignment and phylogenetic analysis revealed that strain HBQ12 and BJH13 were clustered together belonging to fowl adenoviruses D species and serotyped as FAdV-11, whereas strain JSJ13 was classified into fowl adenoviruses C species and serotyped as FAdV-4. To our knowledge, this is the first report of FAdV-4 strain circulating in China. The pathogenicity test showed that mortality for chickens infected with HBQ12 and JSJ13 within 21 days post infection (dpi) was 8.6% and 28.6%, respectively. Necropsy displayed mild or severe hepatitis and hydropericardium at 3 and 5 dpi as well as dead chickens. Viral DNA was detected in almost all tissues sampled from dead chickens. These results revealed that fowl adenovirus strains HBQ12 and JSJ13 are capable of causing IBH and HPS in chickens, indicating that preventive measures against FAdV infection on poultry farms should be implemented in China. PMID:26167857

  18. The effect of cell disruption techniques and chaotropic agents on the downstream purification process of mecasermin produced as inclusion body in E. coli.

    PubMed

    Haddad, Leila; Babaeipour, Valiollah; Mofid, Mohammad Reza

    2015-01-01

    The isolation of the target protein from inclusion bodies (IBs) is a preliminary step to increase protein titer and to maintain its biological activity. In the present study, the effects of various cell lysis methods and the expression temperature was investigated on the improvement of the subsequent purification steps of mecasermin produced in IB. We also investigated the solubilization profile of the top-notch IB in 6 M guanidine hydrochloride (Gdn-HCl) and 8 M urea at different pH ranges. Mecasermin was expressed at various temperatures (25, 28, 30, and 37 °C) and the Escherichia coli cells were lysed by different cell lysis methods. The purity and quality of harvested IBs was evaluated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Finally, mecasermin was refolded and purified using gel filtration chromatography. The profile of SDS-PAGE analysis showed higher quality and purity after application of sonication coupled with lysozyme pretreatment for expressed mecasermin at 37 °C. Besides, from dithiothreitol application in washing step, we achieved a manifold enriched secondary IB for further purification of mecasermin. Mecasermin exhibited optimized solubility in 6 M Gdn-HCl at pH of 5.4. The findings of this study indicate an important role for cell disruption techniques to efficient purification of mecasermin. The study presents the most efficient techniques for improvement of downstream purification of mecasermin. PMID:26779275

  19. Genotype-Phenotype studies of VCP-associated Inclusion Body Myopathy with Paget Disease of Bone and/or Frontotemporal Dementia

    PubMed Central

    Mehta, Sarju G.; Khare, Manaswitha; Ramani, Rupal; Watts, Giles D. J.; Simon, Mariella; Osann, Kathryn E.; Donkervoort, Sandra; Dec, Eric; Nalbandian, Angele; Platt, Julia; Pasquali, Marzia; Wang, Annabel; Mozaffar, Tahseen; Smith, Charles D.; Kimonis, Virginia E.

    2012-01-01

    VCP disease associated with Inclusion body myopathy, Paget disease of the bone and frontotemporal dementia is a progressive autosomal dominant disorder caused by mutations in Valosin containing protein gene. To establish genotype-phenotype correlations we analyzed clinical and biochemical markers from a database of 190 members in 27 families harboring ten missense mutations. Individuals were grouped into three categories: symptomatic, presymptomatic carriers and non-carriers. The symptomatic families were further divided into ten groups based on their VCP mutations. There was marked intra and inter-familial variation; and significant genotype-phenotype correlations were difficult because of small numbers. Nevertheless when comparing the two most common mutations, R155C mutation was found to be more severe, with earlier onset of myopathy and Paget (p=0.03). Survival analysis of all subjects revealed an average life span after diagnosis of myopathy and Paget of 18 and 19 years respectively, and after dementia only 6 years. R155C had a reduced survival compared to the R155H mutation (p=0.03). We identified amyotrophic lateral sclerosis (ALS) in thirteen individuals (8.9%) and Parkinson’s disease in five individuals (3%); however there was no genotypic correlation. This study represents the largest dataset of patients with VCP disease and expands our understanding of natural history and provides genotype-phenotype correlations in this unique disease. PMID:22909335

  20. The effect of cell disruption techniques and chaotropic agents on the downstream purification process of mecasermin produced as inclusion body in E. coli

    PubMed Central

    Haddad, Leila; Babaeipour, Valiollah; Mofid, Mohammad Reza

    2015-01-01

    The isolation of the target protein from inclusion bodies (IBs) is a preliminary step to increase protein titer and to maintain its biological activity. In the present study, the effects of various cell lysis methods and the expression temperature was investigated on the improvement of the subsequent purification steps of mecasermin produced in IB. We also investigated the solubilization profile of the top-notch IB in 6 M guanidine hydrochloride (Gdn-HCl) and 8 M urea at different pH ranges. Mecasermin was expressed at various temperatures (25, 28, 30, and 37 °C) and the Escherichia coli cells were lysed by different cell lysis methods. The purity and quality of harvested IBs was evaluated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Finally, mecasermin was refolded and purified using gel filtration chromatography. The profile of SDS-PAGE analysis showed higher quality and purity after application of sonication coupled with lysozyme pretreatment for expressed mecasermin at 37 °C. Besides, from dithiothreitol application in washing step, we achieved a manifold enriched secondary IB for further purification of mecasermin. Mecasermin exhibited optimized solubility in 6 M Gdn-HCl at pH of 5.4. The findings of this study indicate an important role for cell disruption techniques to efficient purification of mecasermin. The study presents the most efficient techniques for improvement of downstream purification of mecasermin. PMID:26779275

  1. Polymorphism in the TOMM40 gene modifies the risk of developing sporadic inclusion body myositis and the age of onset of symptoms.

    PubMed

    Mastaglia, F L; Rojana-udomsart, A; James, I; Needham, M; Day, T J; Kiers, L; Corbett, J A; Saunders, A M; Lutz, M W; Roses, A D

    2013-12-01

    A polyT repeat in an intronic polymorphism (rs10524523) in the TOMM40 gene, which encodes an outer mitochondrial membrane translocase involved in the transport of amyloid-β and other proteins into mitochondria, has been implicated in Alzheimer's disease and APOE-TOMM40 genotypes have been shown to modify disease risk and age at onset of symptoms. Because of the similarities between Alzheimer's disease and sporadic inclusion body myositis (s-IBM), and the importance of amyloid-β and mitochondrial changes in s-IBM, we investigated whether variation in poly-T repeat lengths in rs10524523 also influence susceptibility and age at onset in a cohort of 90 Caucasian s-IBM patients (55 males; age 69.1 ± 9.6). In carriers of APOE ε3/ε3 or ε3/ε4, genotypes with a very long (VL) poly-T repeat were under-represented in s-IBM compared to controls and were associated with a later age at symptom onset, suggesting that these genotypes may be protective. Our study is the first to suggest that polymorphisms in genes controlling mitochondrial function can influence susceptibility to s-IBM and have disease modifying effects. However, further studies in other s-IBM populations are needed to confirm these findings, as well as expression studies of different TOMM40 alleles in muscle tissue. PMID:24103330

  2. Composition of the Cutaneous Bacterial Community in Japanese Amphibians: Effects of Captivity, Host Species, and Body Region.

    PubMed

    Sabino-Pinto, Joana; Bletz, Molly Catherine; Islam, Mohammed Mafizul; Shimizu, Norio; Bhuju, Sabin; Geffers, Robert; Jarek, Michael; Kurabayashi, Atsushi; Vences, Miguel

    2016-08-01

    The cutaneous microbiota plays a significant role in the biology of their vertebrate hosts, and its composition is known to be influenced both by host and environment, with captive conditions often altering alpha diversity. Here, we compare the cutaneous bacterial communities of 61 amphibians (both wild and captive) from Hiroshima, Japan, using high-throughput amplicon sequencing of a segment of the 16S rRNA gene. The majority of these samples came from a captive breeding facility at Hiroshima University where specimens from six species are maintained under highly standardized conditions for several generations. This allowed to identify host effects on the bacterial communities under near identical environmental conditions in captivity. We found the structure of the cutaneous bacterial community significantly differing between wild and captive individuals of newts, Cynops pyrrhogaster, with a higher alpha diversity found in the wild individuals. Community structure also showed distinct patterns when comparing different species of amphibians kept under highly similar conditions, revealing an intrinsic host effect. Bacterial communities of dorsal vs. ventral skin surfaces did not significantly differ in most species, but a trend of higher alpha diversity on the ventral surface was found in Oriental fire-bellied toads, Bombina orientalis. This study confirms the cutaneous microbiota of amphibians as a highly dynamic system influenced by a complex interplay of numerous factors. PMID:27278778

  3. The broad-spectrum antiviral compound ST-669 restricts chlamydial inclusion development and bacterial growth and localizes to host cell lipid droplets within treated cells.

    PubMed

    Sandoz, Kelsi M; Valiant, William G; Eriksen, Steven G; Hruby, Dennis E; Allen, Robert D; Rockey, Daniel D

    2014-07-01

    Novel broad-spectrum antimicrobials are a critical component of a strategy for combating antibiotic-resistant pathogens. In this study, we explored the activity of the broad-spectrum antiviral compound ST-669 for activity against different intracellular bacteria and began a characterization of its mechanism of antimicrobial action. ST-669 inhibits the growth of three different species of chlamydia and the intracellular bacterium Coxiella burnetii in Vero and HeLa cells but not in McCoy (murine) cells. The antichlamydial and anti-C. burnetii activity spectrum was consistent with those observed for tested viruses, suggesting a common mechanism of action. Cycloheximide treatment in the presence of ST-669 abrogated the inhibitory effect, demonstrating that eukaryotic protein synthesis is required for tested activity. Immunofluorescence microscopy demonstrated that different chlamydiae grow atypically in the presence of ST-669, in a manner that suggests the compound affects inclusion formation and organization. Microscopic analysis of cells treated with a fluorescent derivative of ST-669 demonstrated that the compound localized to host cell lipid droplets but not to other organelles or the host cytosol. These results demonstrate that ST-669 affects intracellular growth in a host-cell-dependent manner and interrupts proper development of chlamydial inclusions, possibly through a lipid droplet-dependent process. PMID:24777097

  4. NMR structure note: Structure of the Membrane Protein MerF, a Bacterial Mercury Transporter, Improved by the Inclusion of Chemical Shift Anisotropy Constraints

    PubMed Central

    Tian, Ye; Lu, George J.; Marassi, Francesca M.; Opella, Stanley J.

    2014-01-01

    SUMMARY MerF is a mercury transport membrane protein from the bacterial mercury detoxification system. By performing a solid-state INEPT experiment and measuring chemical shift anisotropy frequencies in aligned samples, we are able to improve on the accuracy and precision of the initial structure that we presented. MerF has four N-terminal and eleven C-terminal residues that are mobile and unstructured in phospholipid bilayers. The structure presented here has average pairwise RMSDs of 1.78 Å for heavy atoms and 0.92 Å for backbone atoms. PMID:25103921

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

    PubMed

    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. PMID:24128692

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

    PubMed

    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. PMID:26632503

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

    PubMed

    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. PMID:26232416

  8. Overexpressed Arabidopsis Annexin4 accumulates in inclusion bodylike structures

    PubMed Central

    Khachatoorian, Careen; Ramirez, Rigoberto A.; Hernandez, Fernando; Serna, Raphael; Kwok, Ernest Y.

    2015-01-01

    Large protein complexes form in the cytosol of prokaryotes and eukaryotes as assemblies of functional enzymes or aggregates of misfolded proteins. Their roles in the cell range from critical components of metabolism to disease-causing agents. We have observed a novel structure in the cells of transgenic Arabidopsis thaliana that appears to be a form of inclusion body. These long, spindle-shaped structures form when Arabidopsis are transformed to express high levels of the protein Annexin4 fused to a fluorescent protein. These structures, previously named darts, are visible in all cells of the plant throughout development. Darts take on a variety of morphologies including rings and figure-eights. These structures are not associated with the endomembrane system and are not membrane bounded. Darts appear to be insoluble aggregates of protein analogous to bacterial inclusion bodies and eukaryotic aggresomes. Similar structures have not been observed in untransformed plants, suggesting darts are artifacts of transgenic overexpression. PMID:25818562

  9. Characterization of the bacterial community associated with body wall lesions of Tripneustes gratilla (Echinoidea) using culture-independent methods.

    PubMed

    Becker, Pierre T; Gillan, David C; Eeckhaut, Igor

    2009-02-01

    The bacterial community associated with skin lesions of the sea urchin Tripneustes gratilla was investigated using 16S ribosomal RNA gene cloning and fluorescent in situ hybridization (FISH). All clones were classified in the Alphaproteobacteria, Gammaproteobacteria and Cytophaga-Flexibacter-Bacteroides (CFB) bacteria. Most of the Alphaproteobacteria were related to the Roseobacter lineage and to bacteria implicated in marine diseases. The majority of the Gammaproteobacteria were identified as Vibrio while CFB represented only 9% of the total clones. FISH analyses showed that Alphaproteobacteria, CFB bacteria and Gammaproteobacteria accounted respectively for 43%, 38% and 19% of the DAPI counts. The importance of the methods used is emphasized. PMID:19041326

  10. Two major histocompatibility complex haplotypes influence susceptibility to sporadic inclusion body myositis: critical evaluation of an association with HLA-DR3.

    PubMed

    Price, P; Santoso, L; Mastaglia, F; Garlepp, M; Kok, C C; Allcock, R; Laing, N

    2004-11-01

    Previous studies of sporadic inclusion body myositis (sIBM) have shown a strong association with HLA-DR3 and other components of the 8.1 ancestral haplotype (AH) (HLA-A1, B8, DR3), where the susceptibility locus has been mapped to the central major histocompatibility complex (MHC) region between HLA-DR and C4. Here, the association with HLA-DR3 and other genes in the central MHC and class II region was further investigated in a group of 42 sIBM patients and in an ethnically similar control group (n = 214), using single-nucleotide polymorphisms and microsatellite screening. HLA-DR3 (marking DRB1*0301 in Caucasians) was associated with sIBM (Fisher's test). However, among HLA-DR3-positive patients and controls, carriage of HLA-DR3 without microsatellite and single-nucleotide polymorphism alleles of the 8.1AH (HLA-A1, B8, DRB3*0101, DRB1*0301, DQB1*0201) was marginally less common in patients. Patients showed no increase in carriage of the 18.2AH (HLA-A30, B18, DRB3*0202, DRB1*0301, DQB1*0201) or HLA-DR3 without the central MHC of the 8.1AH, further arguing against HLA-DRB1 as the direct cause of susceptibility. Genes between HLA-DRB1 and HOX12 require further investigation. BTL-II lies in this region and is expressed in muscle. Carriage of allele 2 (exon 6) was more common in patients. BTL-II(E6)*2 is characteristic of the 35.2AH (HLA-A3, B35, DRB1*01) in Caucasians and HLA-DR1, BTL-II(E6)*2, HOX12*2, RAGE*2 was carried by several patients. The 8.1AH and 35.2AH may confer susceptibility to sIBM independently or share a critical allele. PMID:15496200

  11. BACE-1, PS-1 and sAPPβ Levels Are Increased in Plasma from Sporadic Inclusion Body Myositis Patients: Surrogate Biomarkers among Inflammatory Myopathies

    PubMed Central

    Catalán-García, Marc; Garrabou, Glòria; Morén, Constanza; Guitart-Mampel, Mariona; Gonzalez-Casacuberta, Ingrid; Hernando, Adriana; Gallego-Escuredo, Jose Miquel; Yubero, Dèlia; Villarroya, Francesc; Montero, Raquel; O-Callaghan, Albert Selva; Cardellach, Francesc; Grau, Josep Maria

    2015-01-01

    Sporadic inclusion body myositis (sIBM) is a rare disease that is difficult to diagnose. Muscle biopsy provides three prominent pathological findings: inflammation, mitochondrial abnormalities and fibber degeneration, represented by the accumulation of protein depots constituted by β-amyloid peptide, among others. We aim to perform a screening in plasma of circulating molecules related to the putative etiopathogenesis of sIBM to determine potential surrogate biomarkers for diagnosis. Plasma from 21 sIBM patients and 20 age- and gender-paired healthy controls were collected and stored at −80°C. An additional population of patients with non-sIBM inflammatory myopathies was also included (nine patients with dermatomyositis and five with polymyositis). Circulating levels of inflammatory cytokines (interleukin [IL]-6 and tumor necrosis factor [TNF]-α), mitochondrial-related molecules (free plasmatic mitochondrial DNA [mtDNA], fibroblast growth factor-21 [FGF-21] and coenzyme-Q10 [CoQ]) and amyloidogenic-related molecules (beta-secretase-1 [BACE-1], presenilin-1 [PS-1], and soluble Aβ precursor protein [sAPPβ]) were assessed with magnetic bead–based assays, real-time polymerase chain reaction, enzyme-linked immunosorbent assay (ELISA) and high-pressure liquid chromatography (HPLC). Despite remarkable trends toward altered plasmatic expression of inflammatory and mitochondrial molecules (increased IL-6, TNF-α, circulating mtDNA and FGF-21 levels and decreased content in CoQ), only amyloidogenic degenerative markers including BACE-1, PS-1 and sAPPβ levels were significantly increased in plasma from sIBM patients compared with controls and other patients with non-sIBM inflammatory myopathies (p < 0.05). Inflammatory, mitochondrial and amyloidogenic degeneration markers are altered in plasma of sIBM patients confirming their etiopathological implication in the disease. Sensitivity and specificity analysis show that BACE-1, PS-1 and sAPPβ represent a good

  12. Blocking monocyte transmigration in in vitro system by an anti-CD99 human antibody in single chain fragment variable (scFv) format. Efficient large scale purification of biological active scFv from inclusion bodies in E. coli expression system

    PubMed Central

    Moricoli, Diego; Muller, William A.; Carbonella, Damiano Cosimo; Balducci, Maria Cristina; Dominici, Sabrina; Fiori, Valentina; Watson, Richard; Weber, Evan; Cianfriglia, Maurizio; Scotlandi, Katia; Magnani, Mauro

    2015-01-01

    Migration of leukocytes into a site of inflammation involves several steps mediated by various families of adhesion molecules. CD99 play a significant role in transendothelial migration (TEM) of leukocytes. Inhibition of TEM by specific monoclonal antibody (mAb) can provide a potent therapeutic approach to treating inflammatory conditions. However, the therapeutic utilization of whole IgG can lead to an inappropriate activation of Fc receptor-expressing cells inducing serious adverse side effects due to cytokine release. In this regard, specific recombinant antibody in single chain variable fragments (scFvs) originated by phage library may offer a solution by affecting TEM function in a safe clinical context. However, this consideration requires large scale production of functional scFv antibodies under GMP conditions and hence, the absence of toxic reagents utilized for the solubilization and refolding steps of inclusion bodies that may discourage industrial application of these antibody fragments. In order to apply the scFv anti-CD99 named C7A in a clinical setting we herein describe an efficient and large scale production of the antibody fragments expressed in E.coli as insoluble protein avoiding gel filtration chromatography approach, and laborious refolding step pre- and post-purification. Using differential salt elution which is a simple, reproducible and effective procedure we are able to separate scFv in monomer format from aggregates. The purified scFv antibody C7A exhibits inhibitory activity comparable to an antagonistic conventional mAb, thus providing an excellent agent for blocking CD99 signalling. Thanks to the original purification protocol that can be extended to other scFvs that are expressed as inclusion bodies in bacterial systems, the scFv anti-CD99 C7A herein described represents the first step towards the construction of new antibody therapeutic. PMID:24798881

  13. Mass determination and estimation of subunit stoichiometry of the bacterial hook-basal body flagellar complex of Salmonella typhimurium by scanning transmission electron microscopy.

    PubMed Central

    Sosinsky, G E; Francis, N R; DeRosier, D J; Wall, J S; Simon, M N; Hainfeld, J

    1992-01-01

    The basal body, a part of the rotary motor of the bacterial flagellum, is a multiprotein assembly that consists of four rings (denoted M, S, P, and L) and an axial rod (denoted R). From analysis of scanning transmission electron microscopy images of hook-basal body preparations isolated from Salmonella typhimurium, we have determined the masses of the basal body and three of its subcomplexes. The mass of the basal body (i.e., the four rings and rod) is 4400 +/- 490 kDa (mean +/- SD; n = 54). The mass of the LPR subcomplex (i.e., L and P rings and the whole rod) is 2600 +/- 380 kDa (n = 55), that of the L and P rings and the distal part of the rod is 2100 +/- 320 kDa (n = 25), and the mass of the L and P ring subcomplex is 1700 +/- 260 kDa (n = 514). These results, together with the masses of the component proteins, indicate that the rings contain approximately 26 subunits each and that the mass of the rod is consistent with a composition of approximately 6 copies each of three of the rod proteins FlgB, FlgC, and FlgF and approximately 26 copies of FlgG as determined by Jones et al. [Jones, C. J., Macnab, R. M., Okino, H. & Aizawa, S.-I. (1990) J. Mol. Biol. 212, 377-387] using quantitative gel electrophoresis. The results of Jones et al., together with ours, account for all proteins in the basal body to within approximately 5% (or 200 kDa). Images PMID:1594581

  14. Inclusive health.

    PubMed

    Maclachlan, Malcolm; Khasnabis, Chapal; Mannan, Hasheem

    2012-01-01

    We propose the concept of Inclusive Health to encapsulate the Health for All ethos; to build on the rights-based approach to health; to promote the idea of inclusion as a verb, where a more proactive approach to addressing distinctive and different barriers to inclusion is needed; and to recognise that new initiatives in human resources for health can offer exciting and innovative ways of healthcare delivery. While Inclusive Education has become a widely recognised and accepted concept, Health for All is still contested, and new thinking is required to develop its agenda in line with contemporary developments. Inclusive Health refers both to who gets health care and to who provides it; and its ethos resonates strongly with Jefferson's assertion that 'there is nothing more unequal, than the equal treatment of unequal people'. We situate the timeliness of the Inclusive Health concept with reference to recent developments in the recognition of the rights of people with disability, in the new guidelines for community-based rehabilitation and in the World Report on Disability. These developments offer a more inclusive approach to health and, more broadly, its inter-connected aspects of wellbeing. A concept which more proactively integrates United Nations conventions that recognise the importance of difference - disability, ethnicity, gender, children - could be of benefit for global healthcare policy and practice. PMID:21895893

  15. Inclusive teaching.

    PubMed

    Billings, Diane M

    2008-07-01

    Inclusive teaching involves being responsive to the diversity represented in the classroom and assisting learners to focus on their culture, attitudes, and beliefs while learning to communicate and collaborate with each other and their patients. PMID:18649804

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

    PubMed Central

    Ersoz, Gulden; Aytacoglu, Barlas Naim; Sucu, Nehir; Tamer, Lulufer; Bayindir, Ismet; Kose, Necmi; Kaya, Ali; Dikmengil, Murat

    2006-01-01

    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 < 0.001). MDA levels were significantly higher in these two groups than

  17. Preschool Inclusion.

    ERIC Educational Resources Information Center

    Cavallaro, Claire C.; Haney, Michele

    This resource guide provides field-tested, research-based strategies for including young children with disabilities in early childhood programs. It is designed to help preservice and in-service educators, Head Start personnel, and child care providers with information to promote inclusion in children's earliest social and educational experiences.…

  18. Macrothrombocytopenia With Döhle Body-Like Granulocyte Inclusions: A Case Report of May-Hegglin Anomaly in a 33-Year-Old White Woman With an Update on the Molecular Findings of MYH9-Related Disease.

    PubMed

    Ruhoy, Steven M; Yates, Amanda

    2016-08-01

    A 33-year-old white woman arrived at the hospital to undergo a hysterectomy due to uterine fibroids. Blood smear review identified macrothrombocytopenia and Döhle body-like cytoplasmic leukocyte inclusions. Genetic testing identified a mutation in exon 39 of the myosin heavy chain gene (MHY9; OMIM 160775), which confirmed the diagnosis of May-Hegglin anomaly. May-Hegglin anomaly is one of a spectrum of MYH9 disorders that also includes Sebastian, Epstein, and Fechtner syndromes. Herein, we describe the clinical and laboratory presentation of a patient with May-Hegglin anomaly and provide an update on the molecular findings and a discussion of the genotypic-phenotypic correlations in this potentially underdiagnosed disorder. PMID:27353381

  19. A Collaborative Group Method of Inclusive Research

    ERIC Educational Resources Information Center

    Bigby, Christine; Frawley, Patsie; Ramcharan, Paul

    2014-01-01

    Background: Funding bodies in Australia and the United Kingdom require research on issues that affect the lives of people with intellectual disability to be inclusive. Debate continues about the nature and benefits of inclusive research, which has become an umbrella term encompassing a broad spectrum of approaches. Method: This study proposes one…

  20. Diversity ? Inclusion: Promoting Integration in Higher Education

    ERIC Educational Resources Information Center

    Tienda, Marta

    2013-01-01

    I argue that enrollment of a diverse student body is but a pragmatic first step toward the broader social goal of inclusion and ask whether motives for campus diversification are aligned with pedagogic goals. I address this question by focusing on inclusion, namely, organizational strategies and practices that promote meaningful social and…

  1. Singing and social inclusion.

    PubMed

    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

  2. Singing and social inclusion

    PubMed Central

    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

  3. Limits to Inclusion

    ERIC Educational Resources Information Center

    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,…

  4. The crack-inclusion interaction problem

    NASA Technical Reports Server (NTRS)

    Liu, X.-H.; Erdogan, F.

    1986-01-01

    The general plane elastostatic problem of interaction between a crack and an inclusion is considered. The Green's functions for a pair of dislocations and a pair of concentrated body forces are used to generate the crack and the inclusion. Integral equations are obtained for a line crack and an elastic line inclusion having an arbitrary relative orientation and size. The nature of stress singularity around the end points of rigid and elastic inclusions is described and three special cases of this intersection problem are studied. The problem is solved for an arbitrary uniform stress state away from the crack-inclusion region. The nonintersecting crack-inclusion problem is considered for various relative size, orientation, and stiffness parameters, and the stress intensity factors at the ends of the inclusion and the crack are calculated. For the crack-inclusion intersection case, special stress intensity factors are defined and are calculated for various values of the parameters defining the relative size and orientation of the crack and the inclusion and the stiffness of the inclusion.

  5. The crack-inclusion interaction problem

    NASA Technical Reports Server (NTRS)

    Xue-Hui, L.; Erdogan, F.

    1984-01-01

    The general plane elastostatic problem of interaction between a crack and an inclusion is considered. The Green's functions for a pair of dislocations and a pair of concentrated body forces are used to generate the crack and the inclusion. Integral equations are obtained for a line crack and an elastic line inclusion having an arbitrary relative orientation and size. The nature of stress singularity around the end points of rigid and elastic inclusions is described and three special cases of this intersection problem are studied. The problem is solved for an arbitrary uniform stress state away from the crack-inclusion region. The nonintersecting crack-inclusion problem is considered for various relative size, orientation, and stiffness parameters, and the stress intensity factors at the ends of the inclusion and the crack are calculated. For the crack-inclusion intersection case, special stress intensity factors are defined and are calculated for various values of the parameters defining the relative size and orientation of the crack and the inclusion and the stiffness of the inclusion.

  6. Basophilic inclusions and neuronal intermediate filament inclusions in amyotrophic lateral sclerosis and frontotemporal lobar degeneration.

    PubMed

    Ito, Hidefumi

    2014-12-01

    Basophilic inclusions (BIs) and neuronal intermediate filament inclusions (NIFIs) are key structures of basophilic inclusion body disease and neuronal intermediate filament inclusion disease (NIFID), respectively. BIs are sharply-defined, oval or crescent neuronal intracytoplasmic inclusions that appear pale blue-gray in color with HE staining and purple in color with Nissl but are stained poorly with silver impregnation techniques. Immunohistochemically BIs are negative for tau, trans-activation response DNA 43 (TDP-43), α-synuclein, neurofilament (NF) and α-internexin, positive for p62, and variably ubiquitinated. Noticeably, BIs are consistently fused in sarcoma (FUS) positive. NIFIs are by definition immuno-positive for class IV IFs including three NF triplet subunit proteins and α-internexin but negative for tau, TDP-43, and α-synuclein. In NIFID cases several types of inclusions have been identified. Among them, hyaline conglomerate-like inclusions are the only type that meets the above immunohistochemical features of NIFIs. This type of inclusion appears upon HE staining as multilobulated, faintly eosinophilic or pale amphophilic spherical masses with a glassy appearance. These hyaline conglomerates appear strongly argyrophilic, and robustly and consistently immuno-positive for IFs. In contrast, this type of inclusion shows no or only occasional dot-like FUS immunoreactivity. Therefore, BIs and NIFIs are distinct from each other in terms of morphological, tinctorial and immunohistochemical features. However, basophilic inclusion body disease (BIBD) and NIFID are difficult to differentiate clinically. Moreover, Pick body-like inclusions, the predominant type of inclusions seen in NIFID, are considerably similar to the BIs of BIBD in that this type of inclusion is basophilic, poorly argyrophilic, negative for IFs and intensely immuno-positive for FUS. As BIBD and NIFID share FUS accumulation as the most prominent molecular pathology, whether these two

  7. Bacterial Sialidase

    NASA Technical Reports Server (NTRS)

    2004-01-01

    Data shows that elevated sialidase in bacterial vaginosis patients correlates to premature births in women. Bacterial sialidase also plays a significant role in the unusual colonization of Pseudomonas aeruginosa in cystic fibrosis patients. Crystals of Salmonella sialidase have been reproduced and are used for studying the inhibitor-enzyme complexes. These inhibitors may also be used to inhibit a trans-sialidase of Trypanosome cruzi, a very similar enzyme to bacterial sialidase, therefore preventing T. cruzi infection, the causitive agent of Chagas' disease. The Center for Macromolecular Crystallography suggests that inhibitors of bacterial sialidases can be used as prophylactic drugs to prevent bacterial infections in these critical cases.

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

    PubMed Central

    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. PMID:26887227

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

    PubMed

    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. PMID:26887227

  10. Shock Re-equilibration of Fluid Inclusions

    NASA Technical Reports Server (NTRS)

    Madden, M. E. Elwood; Horz, F.; Bodnar, R. J.

    2004-01-01

    Fluid inclusions (microscopic volumes of fluid trapped within minerals as they precipitate) are extremely common in terrestrial minerals formed under a wide range of geological conditions from surface evaporite deposits to kimberlite pipes. While fluid inclusions in terrestrial rocks are nearly ubiquitous, only a few fluid inclusion-bearing meteorites have been documented. The scarcity of fluid inclusions in meteoritic materials may be a result of (a) the absence of fluids when the mineral was formed on the meteorite parent body or (b) the destruction of fluid inclusions originally contained in meteoritic materials by subsequent shock metamorphism. However, the effects of impact events on pre-existing fluid inclusions trapped in target and projectile rocks has received little study. Fluid inclusions trapped prior to the shock event may be altered (re-equilibrated) or destroyed due to the high pressures, temperatures, and strain rates associated with impact events. By examining the effects of shock deformation on fluid inclusion properties and textures we may be able to better constrain the pressure-temperature path experienced by terrestrial and meteoritic shocked materials and also gain a clearer understanding of why fluid inclusions are rarely found in meteorite samples.

  11. Ribonucleotides in Bacterial DNA

    PubMed Central

    Schroeder, Jeremy W.; Randall, Justin R.; Matthews, Lindsay A.; Simmons, Lyle A.

    2014-01-01

    In all living cells, DNA is the storage medium for genetic information. Being quite stable, DNA is well-suited for its role in storage and propagation of information, but RNA is also covalently included in DNA through various mechanisms. Recent studies also demonstrate useful aspects of including ribonucleotides in the genome during repair. Therefore, our understanding of the consequences of RNA inclusion into bacterial genomic DNA is just beginning, but with its high frequency of occurrence the consequences and potential benefits are likely to be numerous and diverse. In this review, we discuss the processes that cause ribonucleotide inclusion in genomic DNA, the pathways important for ribonucleotide removal and the consequences that arise should ribonucleotides remain nested in genomic DNA. PMID:25387798

  12. Bacterial Proteasomes

    PubMed Central

    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. PMID:26488274

  13. Fluid Inclusion Gas Analysis

    DOE Data Explorer

    Dilley, Lorie

    2013-01-01

    Fluid inclusion gas analysis for wells in various geothermal areas. Analyses used in developing fluid inclusion stratigraphy for wells and defining fluids across the geothermal fields. Each sample has mass spectrum counts for 180 chemical species.

  14. Inclusive Education in Bangladesh

    ERIC Educational Resources Information Center

    Ahsan, Mohammad Tariq; Burnip, Lindsay

    2007-01-01

    This article reports on inclusive education in Bangladesh for children with special needs. Bangladesh is not behind other developed countries in enacting laws and declarations in favour of inclusive education, but a lack of resources is the main barrier in implementing inclusive education. Special education and integrated education models exist in…

  15. Inclusion in Middle Tennessee

    ERIC Educational Resources Information Center

    Hayes, Brandalyn; Ashley, Mandi; Salter, Derrick

    2013-01-01

    The overall purpose of this study was to provide school districts within Tennessee with more research about how weekly hours of inclusion impact student achievement. Specifically, researchers examined which models of inclusion were in use in two school districts in Tennessee, administrators' and teachers' perceptions of inclusion, and whether or…

  16. Towards Inclusive Schooling

    ERIC Educational Resources Information Center

    Gafoor, K. Abdul

    2010-01-01

    Social inclusion is the process that will enable every person in society to participate in normal activities of societies they live in, including education, employment, public services and social recreational activities. For the development of an inclusive society, preparation of younger generation also needs to be inclusive. Our schools must…

  17. Supporting Inclusive Practice

    ERIC Educational Resources Information Center

    Knowles, Gianna

    2006-01-01

    Written to support all teaching and learning staff in developing good inclusive practice, this book provides knowledge and understanding about a range of inclusion issues, such as what an inclusive school might look like and practical guidance on supporting the development of such a school. It also explores issues surrounding: (1) Ethnicity; (2)…

  18. Footstep towards Inclusive Education

    ERIC Educational Resources Information Center

    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…

  19. Disruption of Type I Interferon Signaling by the Nonstructural Protein of Severe Fever with Thrombocytopenia Syndrome Virus via the Hijacking of STAT2 and STAT1 into Inclusion Bodies

    PubMed Central

    Ning, Yun-Jia; Feng, Kuan; Min, Yuan-Qin; Cao, Wu-Chun; Wang, Manli; Deng, Fei; Hu, Zhihong

    2015-01-01

    ABSTRACT The type I interferon (IFN) system, including IFN induction and signaling, is the critical component of the host defense line against viral infection, which, in turn, is also a vulnerable target for viral immune evasion. Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging bunyavirus. Previous data have shown that SFTSV can interfere with the early induction of type I IFNs through targeting host kinases TBK1/IKKε. In this study, we demonstrated that SFTSV also can suppress type I IFN-triggered signaling and interferon-stimulated gene (ISG) expression. Interestingly, we observed the significant inhibition of IFN signaling in cells transfected with the plasmids encoding the nonstructural protein (NSs) but not the nucleocapsid protein (NP), indicating the role of NSs as an antagonist of IFN signaling. Furthermore, coimmunoprecipitation (Co-IP) and pulldown assays indicated that NSs interacts with the cellular signal transducer and activator of transcription 2 (STAT2), and the DNA-binding domain of STAT2 may contribute to the NSs-STAT2 interaction. Combined with confocal microscopy analyses, we demonstrated that NSs sequesters STAT2 and STAT1 into viral inclusion bodies (IBs) and impairs IFN-induced STAT2 phosphorylation and nuclear translocation of both STATs, resulting in the inhibition of IFN signaling and ISG expression. SFTSV NSs-mediated hijacking of STATs in IBs represents a novel mechanism of viral suppression of IFN signaling, highlighting the role of viral IBs as the virus-built “jail” sequestering some crucial host factors and interfering with the corresponding cellular processes. IMPORTANCE SFTSV is an emerging bunyavirus which can cause a severe hemorrhagic fever-like disease with high case fatality rates in humans, posing a serious health threat. However, there are no specific antivirals available, and the pathogenesis and virus-host interactions are largely unclear. Here, we demonstrated that SFTSV can inhibit type I IFN

  20. Bacterial Keratitis

    MedlinePlus

    ... very quickly, and if left untreated, can cause blindness. The bacteria usually responsible for this type of ... to intense ultraviolet radiation exposure, e.g. snow blindness or welder's arc eye). Next Bacterial Keratitis Symptoms ...

  1. Towards Inclusive Schooling.

    ERIC Educational Resources Information Center

    Ainscow, Mel

    1997-01-01

    Uses classroom vignettes to examine reasons why schools in the United Kingdom are not yet generally successful in including students with disabilities and suggests simple ways that ordinary teachers can implement inclusive practices. These include the importance of teamwork, a school climate which encourages inclusive practices, and teacher…

  2. Conclusions on Inclusion

    ERIC Educational Resources Information Center

    Fink, John

    2004-01-01

    An inclusion classroom is defined as one that is tasked with mainstreaming special education students into a population of general education students. In this brief article, the author, a high school mathematics teacher, shares his personal experiences in teaching in an inclusion classroom. A primary focus is his relationship with the special…

  3. The Inclusion Facilitator's Guide

    ERIC Educational Resources Information Center

    Jorgensen, Cheryl M.; Schuh, Mary C.; Nisbet, Jan

    2005-01-01

    Inclusion facilitators are educators who do more than teach children with disabilities--they advocate for change in schools and communities, sparking a passion for inclusion in teachers, administrators, and families and giving them the practical guidance they need to make it work. This is an essential new role in today's schools, and this guide…

  4. Teachers' Attitudes toward Inclusion

    ERIC Educational Resources Information Center

    Elhoweris, Hala; Alsheikh, Negmeldin

    2004-01-01

    The purpose of this study was to (a) investigate current teachers' attitudes toward inclusion, and to (b) explore possible difference in the general and special education teachers' attitudes toward inclusion of students with disabilities in the integrated education classroom. A total of 10 participants from a large mid-western state university…

  5. Fluid inclusion petrography

    NASA Astrophysics Data System (ADS)

    Van den Kerkhof, Alfons M.; Hein, Ulrich F.

    2001-01-01

    A procedure of fluid inclusion studies is proposed with emphasis on the criteria of selecting fluid inclusions for detailed (microthermometry and spectroscopic) analysis. An overview of descriptive and genetic classifications of fluid inclusions in single crystals and in massive rocks is given with the intention of further differentiating the commonly used terms 'primary' and 'secondary' fluid inclusions. Some principles of fluid inclusion modification are explained. Cathodoluminescence (CL) studies of quartz with the optical high-power CL-microscope and the electron microprobe provided with a CL detector are an important help in 'fluid petrography'. CL textures are subdivided in primary, growth textures and a wide variety of secondary microtextures, which are in part induced by fluid inclusions. The latter is grouped in textures indicative of local lower crystal order (increasing defect structures) and microtextures indicative of local quartz healing (reduction of the defect structures). Microtextures showing the genetic relationship between fluid inclusions and the host mineral provide information about the possible post-entrapment changes of fluid inclusions and therewith testify their geological relevance.

  6. Index for Inclusion

    ERIC Educational Resources Information Center

    Smith, Allister

    2005-01-01

    Index for Inclusion is a programme to assist in developing learning and participation in schools. It was written by Tony Booth and Mel Ainscow from the Centre for Studies on Inclusive Education, UK. Central Normal School was pleased to have the opportunity to trial this programme.

  7. What Counts as Inclusion?

    ERIC Educational Resources Information Center

    Walton, E.; Nel, N.

    2012-01-01

    In the years since the publication in South Africa of White Paper Six: Special needs education (Department of Education (DoE) 2001) various schools in the state and independent sectors have begun to implement inclusive policies and practices. With reference to the Guidelines for full-service/inclusive schools issued in 2009, and by discussing a…

  8. Understanding Inclusion in Cyprus

    ERIC Educational Resources Information Center

    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…

  9. Inclusive Services Innovation Configuration

    ERIC Educational Resources Information Center

    Holdheide, Lynn R.; Reschly, Daniel J.

    2011-01-01

    Teacher preparation to deliver inclusive services to students with disabilities is increasingly important because of changes in law and policy emphasizing student access to, and achievement in, the general education curriculum. This innovation configuration identifies the components of inclusive services that should be incorporated in teacher…

  10. Jet inclusive cross sections

    SciTech Connect

    Del Duca, V.

    1992-11-01

    Minijet production in jet inclusive cross sections at hadron colliders, with large rapidity intervals between the tagged jets, is evaluated by using the BFKL pomeron. We describe the jet inclusive cross section for an arbitrary number of tagged jets, and show that it behaves like a system of coupled pomerons.

  11. Equity and Inclusion in Physical Education PLC

    ERIC Educational Resources Information Center

    Evans, John

    2014-01-01

    Physical Educationalists in many western and westernised societies across the globe are facing new challenges as system wide changes take place increasing the role of private bodies (e.g. Academy trusts) in the delivery of school based education. This reflective and rather personal paper considers the place and meaning of "inclusion" and…

  12. In vivo deuteration of a native bacterial biopolymer for structural elucidation using SANS

    NASA Astrophysics Data System (ADS)

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

    2004-07-01

    In order to facilitate future structural studies, biodeuteration of bacterial polyhydroxyalkanoates (PHAs) was investigated. We report here the in vivo deuteration of poly 3-hydroxyoctanoate (PHO) produced by its native host, the bacterium Pseudomonas oleovorans. Bacterial biomass was produced in bioreactor studies by growth on hydrogenated substrates and PHO was subsequently produced intracellularly (10-20% w/w) during batch fed growth on deuterated octanoic acid under oxygen limitation. GC-MS analyses of the PHO demonstrated that 13 of the 15 hydrogen atoms had been replaced with deuterium (except in position 3), the remaining two hydrogen presumably being derived from water. A SANS contrast variation study was conducted on whole cells and the results indicate the potential to discriminate inclusion bodies formed from deuterated precursor from an otherwise hydrogenated background.

  13. Cytomegalic inclusion disease in the east African headgehog.

    PubMed

    Karstad, L

    1975-04-01

    Cytomegaly with intranuclear inclusion bodies was found in the duct epithelium of the salivary glands of five of ten hedgehogs (Atelerix albiventris). Cytomegalovirus infection was presumed to be the cause. PMID:167206

  14. Fluid inclusion geothermometry

    USGS Publications Warehouse

    Cunningham, C.G., Jr.

    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. Bacterial Immunity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A variety of bacterial agents reside in and around the environment that can cause illness and death in a poultry flock. Many cause disseminated disease while others exert more local effects such as the respiratory or gastrointestinal tract. The host, for our current purposes the laying hen, has de...

  16. Bacterial ratchet motors

    PubMed Central

    Di Leonardo, R.; Angelani, L.; Dell’Arciprete, D.; Ruocco, G.; Iebba, V.; Schippa, S.; Conte, M. P.; Mecarini, F.; De Angelis, F.; Di Fabrizio, E.

    2010-01-01

    Self-propelling bacteria are a nanotechnology dream. These unicellular organisms are not just capable of living and reproducing, but they can swim very efficiently, sense the environment, and look for food, all packaged in a body measuring a few microns. Before such perfect machines can be artificially assembled, researchers are beginning to explore new ways to harness bacteria as propelling units for microdevices. Proposed strategies require the careful task of aligning and binding bacterial cells on synthetic surfaces in order to have them work cooperatively. Here we show that asymmetric environments can produce a spontaneous and unidirectional rotation of nanofabricated objects immersed in an active bacterial bath. The propulsion mechanism is provided by the self-assembly of motile Escherichia coli cells along the rotor boundaries. Our results highlight the technological implications of active matter’s ability to overcome the restrictions imposed by the second law of thermodynamics on equilibrium passive fluids. PMID:20457936

  17. Melt inclusions: Chapter 6

    USGS Publications Warehouse

    Audétat A.; Lowenstern, J. B.

    2014-01-01

    Melt inclusions are small droplets of silicate melt that are trapped in minerals during their growth in a magma. Once formed, they commonly retain much of their initial composition (with some exceptions) unless they are re-opened at some later stage. Melt inclusions thus offer several key advantages over whole rock samples: (i) they record pristine concentrations of volatiles and metals that are usually lost during magma solidification and degassing, (ii) they are snapshots in time whereas whole rocks are the time-integrated end products, thus allowing a more detailed, time-resolved view into magmatic processes (iii) they are largely unaffected by subsolidus alteration. Due to these characteristics, melt inclusions are an ideal tool to study the evolution of mineralized magma systems. This chapter first discusses general aspects of melt inclusions formation and methods for their investigation, before reviewing studies performed on mineralized magma systems.

  18. Minority Inclusion Programs.

    ERIC Educational Resources Information Center

    Gelfand, M. David

    1995-01-01

    Asserts that the United States has a long, sad history of discrimination against minority rights. Discusses the origins and history of minority inclusion or affirmative action programs. Includes a special report on the "English Only" movement. (CFR)

  19. Limitations of inclusive fitness.

    PubMed

    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. PMID:24277847

  20. Limitations of inclusive fitness

    PubMed Central

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

    2013-01-01

    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. PMID:24277847

  1. [Bacterial vaginosis].

    PubMed

    Romero Herrero, Daniel; Andreu Domingo, Antonia

    2016-07-01

    Bacterial vaginosis (BV) is the main cause of vaginal dysbacteriosis in the women during the reproductive age. It is an entity in which many studies have focused for years and which is still open for discussion topics. This is due to the diversity of microorganisms that cause it and therefore, its difficult treatment. Bacterial vaginosis is probably the result of vaginal colonization by complex bacterial communities, many of them non-cultivable and with interdependent metabolism where anaerobic populations most likely play an important role in its pathogenesis. The main symptoms are an increase of vaginal discharge and the unpleasant smell of it. It can lead to serious consequences for women, such as an increased risk of contracting sexually transmitted infections including human immunodeficiency virus and upper genital tract and pregnancy complications. Gram stain is the gold standard for microbiological diagnosis of BV, but can also be diagnosed using the Amsel clinical criteria. It should not be considered a sexually transmitted disease but it is highly related to sex. Recurrence is the main problem of medical treatment. Apart from BV, there are other dysbacteriosis less characterized like aerobic vaginitis of which further studies are coming slowly but are achieving more attention and consensus among specialists. PMID:27474242

  2. WELDED JACKETED URANIUM BODY

    DOEpatents

    Gurinsky, D.H.

    1958-08-26

    A fuel element is presented for a neutronic reactor and is comprised of a uranium body, a non-fissionable jacket surrounding sald body, thu jacket including a portion sealed by a weld, and an inclusion in said sealed jacket at said weld of a fiux having a low neutron capture cross-section. The flux is provided by combining chlorine gas and hydrogen in the intense heat of-the arc, in a "Heliarc" welding muthod, to form dry hydrochloric acid gas.

  3. Neuronal intranuclear inclusion disease in identical twins.

    PubMed

    Haltia, M; Somer, H; Palo, J; Johnson, W G

    1984-04-01

    A pair of female identical twins exhibited slurred speech, nystagmus, and oculogyral spasms starting at age 11. The patients then had episodic rage, extrapyramidal and lower motor neuron abnormalities, and grand mal seizures, but retained largely normal intelligence, until death at age 21. Severe loss of nigral and craniospinal motor neurons was noted postmortem. Round, eosinophilic, autofluorescent inclusion bodies, 3 to 10 microns in diameter, were observed in the nuclei of most nerve cell types of the central and peripheral nervous systems and retina. Ultrastructurally the inclusions appeared as masses of filaments without a limiting membrane, the constituent filaments having a diameter of 8.5 to 9.5 nm. Histochemical results suggested the presence of proteins with a high content of tryptophan. Four similar cases have been reported previously under various designations. We propose the name neuronal intranuclear inclusion disease for the disorder. PMID:6331275

  4. Bacterial Infections

    MedlinePlus

    ... body will learn to resist them causing antibiotic resistance. Later, you could get or spread an infection that those antibiotics cannot cure. NIH: National Institute of Allergy and Infectious Diseases

  5. Novel inclusion in laser crystals

    SciTech Connect

    Ma Xiaoshan; Wang Siting; Jin Zhongru; Shen Yafang; Chen Jiaguang

    1986-12-01

    In growing alexandrite crystals, a novel inclusion has been found. The inclusions are quantitatively analyzed by an electronic probe and the mechanism for forming inclusions is suggested. In our Bridgman MgF/sub 2/ crystals, the inclusions in <001> direction have also been observed.

  6. Linguistic Diversity and Social Inclusion

    ERIC Educational Resources Information Center

    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…

  7. Bacterial Games

    NASA Astrophysics Data System (ADS)

    Frey, Erwin; Reichenbach, Tobias

    Microbial laboratory communities have become model systems for studying the complex interplay between nonlinear dynamics of evolutionary selection forces, stochastic fluctuations arising from the probabilistic nature of interactions, and spatial organization. Major research goals are to identify and understand mechanisms that ensure viability of microbial colonies by allowing for species diversity, cooperative behavior and other kinds of "social" behavior. A synthesis of evolutionary game theory, nonlinear dynamics, and the theory of stochastic processes provides the mathematical tools and conceptual framework for a deeper understanding of these ecological systems. We give an introduction to the modern formulation of these theories and illustrate their effectiveness, focusing on selected examples of microbial systems. Intrinsic fluctuations, stemming from the discreteness of individuals, are ubiquitous, and can have important impact on the stability of ecosystems. In the absence of speciation, extinction of species is unavoidable, may, however, take very long times. We provide a general concept for defining survival and extinction on ecological time scales. Spatial degrees of freedom come with a certain mobility of individuals. When the latter is sufficiently high, bacterial community structures can be understood through mapping individual-based models, in a continuum approach, onto stochastic partial differential equations. These allow progress using methods of nonlinear dynamics such as bifurcation analysis and invariant manifolds. We conclude with a perspective on the current challenges in quantifying bacterial pattern formation, and how this might have an impact on fundamental research in nonequilibrium physics .

  8. Inclusion Practice Priorities Instrument.

    ERIC Educational Resources Information Center

    Montie, Jo; And Others

    This instrument was developed to assist individuals or teams to review best practice indicators regarding the development of inclusive school communities and to establish priority targets for improvement. The instrument covers three areas: (1) school community issues, (2) team issues, and (3) classroom issues. For each area, there is a review…

  9. In Search of Inclusion

    ERIC Educational Resources Information Center

    Hunter, John; O'Connor, Una

    2006-01-01

    This article provides a context within which other contributions to this issue might be read. It examines the position of special educational needs (SEN) within the evolving continuum of education in Northern Ireland, specifically within the context of educational inclusion. It describes recent changes in educational policy and legislation which…

  10. Inclusion Art Contest

    ERIC Educational Resources Information Center

    Mel, Melisa Dauzat

    2010-01-01

    The purpose of this study was to ease a school system into the transitions that occur when Inclusion is incorporated. An entire Middle School was used to collect data from. The grades in that school were 6th, 7th and 8th grade. The initial intent was to foment a community feeling among the students. The results were completely unexpected as they…

  11. Exploring Inclusive Pedagogy

    ERIC Educational Resources Information Center

    Florian, Lani; Black-Hawkins, Kristine

    2011-01-01

    This paper reports on a study designed to examine teachers' craft knowledge of their practice of "inclusion" in terms of what they do, why and how. The research approach offers an important alternative to studies of students with "additional needs" and the search to articulate the specialist knowledge and skill required to teach them. Through…

  12. Inclusion on the Bookshelf

    ERIC Educational Resources Information Center

    Jackson, Camille

    2009-01-01

    Three decades have passed since federal law mandated inclusion--ending, officially at least, a system that segregated students with disabilities from the rest of the student population. The publishing world has yet to catch up. In children's books, characters with disabilities often inhabit their own separate world, where disability is the only…

  13. Nanotubular Toughening Inclusions

    NASA Technical Reports Server (NTRS)

    Park, Cheol (Inventor); Working, Dennis C. (Inventor); Siochi, Emilie J. (Inventor); Harrison, Joycelyn S. (Inventor)

    2015-01-01

    Conventional toughening agents are typically rubbery materials or small molecular weight molecules, which mostly sacrifice the intrinsic properties of a matrix such as modulus, strength, and thermal stability as side effects. On the other hand, high modulus inclusions tend to reinforce elastic modulus very efficiently, but not the strength very well. For example, mechanical reinforcement with inorganic inclusions often degrades the composite toughness, encountering a frequent catastrophic brittle failure triggered by minute chips and cracks. Thus, toughening generally conflicts with mechanical reinforcement. Carbon nanotubes have been used as efficient reinforcing agents in various applications due to their combination of extraordinary mechanical, electrical, and thermal properties. Moreover, nanotubes can elongate more than 20% without yielding or breaking, and absorb significant amounts of energy during deformation, which enables them to also be an efficient toughening agent, as well as excellent reinforcing inclusion. Accordingly, an improved toughening method is provided by incorporating nanotubular inclusions into a host matrix, such as thermoset and thermoplastic polymers or ceramics without detrimental effects on the matrix's intrinsic physical properties.

  14. Raising Achievement through Inclusion

    ERIC Educational Resources Information Center

    Persson, Elisabeth

    2013-01-01

    In 2007, Swedish authorities introduced open publication of comparisons of students' results at the end of compulsory school. In this study, we investigated a municipality that had succeeded in breaking a negative trend from a bottom position in the ranking in 2007 to a top position in 2010, apparently through inclusive practices. The purpose…

  15. Teaching Inclusion through Alphapoems.

    ERIC Educational Resources Information Center

    Murray, Susan "BOON"

    2000-01-01

    Describes what inclusion is and how people in the helping professions may unintentionally promote exclusion. Describes the author's use of alphapoems in a course to develop college students' affective learning and advocacy toward people with disabilities. Offers examples of students' work and concludes that alphapoems served to further pedagogical…

  16. Optimization of inclusive fitness.

    PubMed

    Grafen, Alan

    2006-02-01

    The first fully explicit argument is given that broadly supports a widespread belief among whole-organism biologists that natural selection tends to lead to organisms acting as if maximizing their inclusive fitness. The use of optimization programs permits a clear statement of what this belief should be understood to mean, in contradistinction to the common mathematical presumption that it should be formalized as some kind of Lyapunov or even potential function. The argument reveals new details and uncovers latent assumptions. A very general genetic architecture is allowed, and there is arbitrary uncertainty. However, frequency dependence of fitnesses is not permitted. The logic of inclusive fitness immediately draws together various kinds of intra-genomic conflict, and the concept of 'p-family' is introduced. Inclusive fitness is thus incorporated into the formal Darwinism project, which aims to link the mathematics of motion (difference and differential equations) used to describe gene frequency trajectories with the mathematics of optimization used to describe purpose and design. Important questions remain to be answered in the fundamental theory of inclusive fitness. PMID:16046225

  17. Treatment of septicaemia and severe bacterial infections in foals with a new cefquinome formulation: a field study.

    PubMed

    Rohdich, Nadja; Zschiesche, Eva; Heckeroth, Anja; Wilhelm, Cornelia; Leendertse, Ietje; Thomas, Emmanuel

    2009-09-01

    A multicentre field study was conducted in accordance with VICH Guideline on Good Clinical Practice (VICH 2000) to confirm the efficacy and safety of a new formulation of cefquinome for the treatment of naturally occurring severe bacterial infections and septicaemia in foals. Thirty-nine foals suffering from severe bacterial infections (such as pneumonia, gastro-enteritis, arthritis, omphalitis, or wound infections) or acute septicaemia were treated twice daily with the test product (1 mg cefquinome/kg body weight) intravenously for three days and then intramuscularly for three to 11 days. Investigators examined the foals daily and scored both systemic and local clinical signs to assess the response to treatment, treatment success and relapses. On the day of inclusion a blood sample was taken from each foal for IgG determination and blood culture. In case of abnormal clinical findings additional samples were taken for bacteriology. Treatment was successful in 87.2% of cases (34 of 39 foals) and no relapses were observed. The average duration of treatment was 7.5 days. At inclusion, bacterial culture was positive in 40.5% (15 out of 37) of the blood cultures. Escherichia coli, Clostridium perfringens and Staphylococcus spp. were the most common isolates and were all susceptible to cefquinome. E. coli predominated in swabs from umbilical and open wound infections, and in rectal swabs E. coli. There was no correlation between IgG at inclusion and study outcome or treatment duration. The test product was very well tolerated by all of the foals following intravenous and intramuscular injection. The cefquinome formulation tested was effective and safe in the treatment of severe bacterial infections and septicaemia in foals under field conditions. PMID:19813447

  18. Cumberland Falls chondritic inclusions: III. Consortium study of relationship to inclusions in Allan Hills 78113 aubrite

    SciTech Connect

    Lipschutz, M.E.; Verkouteren, R.M. ); Sears, D.W.G.; Hasan, F.A. ); Prinz, M.; Weisberg, M.K.; Nehru, C.E.; Delaney, J.S. ); Grossman, L.; Boily, M. )

    1988-07-01

    The authors describe the mineralogy and report contents of Ag, Au, Bi, Cd, Co, Cs, Ga, In, Rb, Sb, Se, Te, Tl, U and Zn determined by RNAA in three primitive chondritic inclusions from the ALH A78113 aubrite. Comparison of these data with those for large, petrologic type 3 chondritic clasts from the Cumberland Falls aubrite and the discovery of small clasts in it like those in ALH A78113 indicate that all constitute a single chondritic suite. They report thermoluminescence data for Cumberland Falls chondritic inclusions and achondritic host. These results, together with mineralogic, major, minor and trace element information, demonstrate that aubrite inclusions represent a different sort of type 3 chondrite, not an LL3 chondrite altered during equilibration with aubrite host. Instead, the aubrite inclusions represent a distinct chondrite class. These inclusions reflect nebular condensation/accretion over a broad redox range and at temperatures relatively high compared with those at which other type 3 chondrites formed. Limited metamorphism and reduction occurred during condensation/accretion, prior to incorporation into aubrite host. During the impact of the chondritic parent body with the aubrite parent body, chondrite fragments were strongly shocked and cooled rapidly. They then mixed with aubrite host, possibly in a regolith, so that these aubrites now represent impact breccias.

  19. Foreign Body Granulomas.

    PubMed

    Molina-Ruiz, Ana M; Requena, Luis

    2015-07-01

    A large list of foreign substances may penetrate the skin and induce a foreign body granulomatous reaction. These particles can enter the skin by voluntary reasons or be caused by accidental inclusion of external substances secondary to cutaneous trauma. In these cases, foreign body granulomas are formed around such disparate substances as starch, cactus bristles, wood splinters, suture material, pencil lead, artificial hair, or insect mouthparts. The purpose of this article is to update dermatologists, pathologists, and other physicians on the most recent etiopathogenesis, clinical presentations, systemic associations, evaluation, and evidence-based management concerning foreign body granulomatous reactions of skin. PMID:26143429

  20. Bacterial Hydrodynamics

    NASA Astrophysics Data System (ADS)

    Lauga, Eric

    2016-01-01

    Bacteria predate plants and animals by billions of years. Today, they are the world's smallest cells, yet they represent the bulk of the world's biomass and the main reservoir of nutrients for higher organisms. Most bacteria can move on their own, and the majority of motile bacteria are able to swim in viscous fluids using slender helical appendages called flagella. Low-Reynolds number hydrodynamics is at the heart of the ability of flagella to generate propulsion at the micrometer scale. In fact, fluid dynamic forces impact many aspects of bacteriology, ranging from the ability of cells to reorient and search their surroundings to their interactions within mechanically and chemically complex environments. Using hydrodynamics as an organizing framework, I review the biomechanics of bacterial motility and look ahead to future challenges.

  1. Case report: epithelial intracytoplasmic herpes viral inclusions associated with an outbreak of duck virus enteritis

    USGS Publications Warehouse

    Barr, B.C.; Jessup, David A.; Docherty, Douglas E.; Lownestine, L.J.

    1992-01-01

    Several muscovy ducks from a free-roaming flock of 65 muscovy and mallard ducks died over a 3-week period. Three muscovy ducks were necropsied. Gross and microscopic changes were compatible with duck virus enteritis, and the virus was isolated. In addition to intranuclear viral inclusion bodies in several tissues, intracytoplasmic inclusion bodies were present in esophageal and cloacal epithelium, By electron microscopy, the membrane-bound intracytoplasmic inclusions were found to contain enveloped herpesvirus, and nuclei contained herpes viral nucleocapsids.

  2. Developing Inclusive Teachers from an Inclusive Curricular Perspective

    ERIC Educational Resources Information Center

    Opertti, Renato; Brady, Jayne

    2011-01-01

    This article defines inclusive education in light of the Education for All agenda. It then describes key considerations for developing inclusive teachers from the perspective of an inclusive curriculum which seeks to address the needs of all learners. It concludes by outlining several key policy discussion areas which must be addressed if…

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

    ERIC Educational Resources Information Center

    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. Inclusive Education in South Korea

    ERIC Educational Resources Information Center

    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…

  5. Brief, Amazing Moments of Inclusion

    ERIC Educational Resources Information Center

    Fialka, Janice

    2005-01-01

    "Real inclusion" of kinds with special needs occurs everywhere, inside the classroom as well as outside. This is a fairly basic principle, however, it is not always easy to achieve. In this article, the author describes how her family have had to "fight" for inclusive education and shares some amazing moments of inclusion with her son Micah.

  6. Bacterial vaginosis.

    PubMed Central

    Spiegel, C A

    1991-01-01

    Bacterial vaginosis (BV) is the most common of the vaginitides affecting women of reproductive age. It appears to be due to an alteration in the vaginal ecology by which Lactobacillus spp., the predominant organisms in the healthy vagina, are replaced by a mixed flora including Prevotella bivia, Prevotella disiens, Porphyromonas spp., Mobiluncus spp., and Peptostreptococcus spp. All of these organisms except Mobiluncus spp. are also members of the endogenous vaginal flora. While evidence from treatment trials does not support the notion that BV is sexually transmitted, recent studies have shown an increased risk associated with multiple sexual partners. It has also been suggested that the pathogenesis of BV may be similar to that of urinary tract infections, with the rectum serving as a reservoir for some BV-associated flora. The organisms associated with BV have also been recognized as agents of female upper genital tract infection, including pelvic inflammatory disease, and the syndrome BV has been associated with adverse outcome of pregnancy, including premature rupture of membranes, chorioamnionitis, and fetal loss; postpartum endometritis; cuff cellulitis; and urinary tract infections. The mechanisms by which the BV-associated flora causes the signs of BV are not well understood, but a role for H2O2-producing Lactobacillus spp. in protecting against colonization by catalase-negative anaerobic bacteria has been recognized. These and other aspects of BV are reviewed. PMID:1747864

  7. How to measure inclusive fitness.

    PubMed

    Creel, S

    1990-09-22

    Although inclusive fitness (Hamilton 1964) is regarded as the basic currency of natural selection, difficulty in applying inclusive fitness theory to field studies persists, a quarter-century after its introduction (Grafen 1982, 1984; Brown 1987). For instance, strict application of the original (and currently accepted) definition of inclusive fitness predicts that no one should ever attempt to breed among obligately cooperative breeders. Much of this confusion may have arisen because Hamilton's (1964) original verbal definition of inclusive fitness was not in complete accord with his justifying model. By re-examining Hamilton's original model, a modified verbal definition of inclusive fitness can be justified. PMID:1979447

  8. Bacterial tyrosinases.

    PubMed

    Claus, Harald; Decker, Heinz

    2006-01-01

    Tyrosinases are nearly ubiquitously distributed in all domains of life. They are essential for pigmentation and are important factors in wound healing and primary immune response. Their active site is characterized by a pair of antiferromagnetically coupled copper ions, CuA and CuB, which are coordinated by six histidine residues. Such a "type 3 copper centre" is the common feature of tyrosinases, catecholoxidases and haemocycanins. It is also one of several other copper types found in the multi-copper oxidases (ascorbate oxidase, laccase). The copper pair of tyrosinases binds one molecule of atmospheric oxygen to catalyse two different kinds of enzymatic reactions: (1) the ortho-hydroxylation of monophenols (cresolase activity) and (2) the oxidation of o-diphenols to o-diquinones (catecholase activity). The best-known function is the formation of melanins from L-tyrosine via L-dihydroxyphenylalanine (L-dopa). The complicated hydroxylation mechanism at the active centre is still not completely understood, because nothing is known about their tertiary structure. One main reason for this deficit is that hitherto tyrosinases from eukaryotic sources could not be isolated in sufficient quantities and purities for detailed structural studies. This is not the case for prokaryotic tyrosinases from different Streptomyces species, having been intensively characterized genetically and spectroscopically for decades. The Streptomyces tyrosinases are non-modified monomeric proteins with a low molecular mass of ca. 30kDa. They are secreted to the surrounding medium, where they are involved in extracellular melanin production. In the species Streptomyces, the tyrosinase gene is part of the melC operon. Next to the tyrosinase gene (melC2), this operon contains an additional ORF called melC1, which is essential for the correct expression of the enzyme. This review summarizes the present knowledge of bacterial tyrosinases, which are promising models in order to get more insights in

  9. Can We Build Inclusion?

    PubMed

    Kirkeby, Inge Mette; Grangaard, Sidse

    2016-01-01

    Inclusion of children with special needs in kindergartens and preschools may be approached from different angles. This paper raises the question of whether the physical framework of kindergartens makes any difference for daily life at the kindergarten at all, and whether it can support inclusion 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 pedagogical practice in general. Twelve interviews were conducted with experienced teachers from twelve different kindergartens with different amounts of space, varying from a ratio of 2.1 m2 play area per child to 5.5 m2. The results indicated that, for a group of children with special needs in particular, 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 on the answers in the interviews, we found support for answering the question in the title in the affirmative; we can build inclusion! This is because the teachers' experience indicated that, if there was sufficient space per child, there were fewer conflicts and the children managed to stay in the same activity for a much longer period. Sufficient space made it possible to divide the children into smaller groups, and use any secluded space. Therefore, it was much easier for other children to include some children with special needs. Accordingly, we can say that, sufficient space per child and an adequate layout and furnishing of the kindergarten is an advantage for all children. This is a clear example of Universal Design in which architectural

  10. Inclusive and Exclusive |Vub|

    SciTech Connect

    Petrella, Antonio; /Ferrara U. /INFN, Ferrara

    2011-11-17

    The current status of the determinations of CKM matrix element |V{sub ub}| via exclusive and inclusive charmless semileptonic B decays is reviewed. The large datasets collected at the B-Factories, and the increased precision of theoretical calculations have allowed an improvement in the determination of |V{sub ub}|. However, there are still significant uncertainties. In the exclusive approach, the most precise measurement of the pion channel branching ratio is obtained by an untagged analysis. This very good precision can be reached by tagged analyses with more data. The problem with exclusive decays is that the strong hadron dynamics can not be calculated from first principles and the determination of the form factor has to rely on light-cone sum rules or lattice QCD calculations. The current data samples allow a comparison of different FF models with data distributions. With further developments on lattice calculations, the theoretical error should shrink to reach the experimental one. The inclusive approach still provides the most precise |V{sub ub}| determinations. With new theoretical calculations, the mild (2.5{sigma}) discrepancy with respect to the |V{sub ub}| value determined from the global UT fit has been reduced. As in the exclusive approach, theoretical uncertainties represent the limiting factor to the precision of the measurement. Reducing the theoretical uncertainties to a level comparable with the statistical error is challenging. New measurements in semileptonic decays of charm mesons could increase the confidence in theoretical calculations and related uncertainties.

  11. Asymmetric inclusion process

    NASA Astrophysics Data System (ADS)

    Reuveni, Shlomi; Eliazar, Iddo; Yechiali, Uri

    2011-10-01

    We introduce and explore the asymmetric inclusion process (ASIP), an exactly solvable bosonic counterpart of the fermionic asymmetric exclusion process (ASEP). In both processes, random events cause particles to propagate unidirectionally along a one-dimensional lattice of n sites. In the ASEP, particles are subject to exclusion interactions, whereas in the ASIP, particles are subject to inclusion interactions that coalesce them into inseparable clusters. We study the dynamics of the ASIP, derive evolution equations for the mean and probability generating function (PGF) of the sites’ occupancy vector, obtain explicit results for the above mean at steady state, and describe an iterative scheme for the computation of the PGF at steady state. We further obtain explicit results for the load distribution in steady state, with the load being the total number of particles present in all lattice sites. Finally, we address the problem of load optimization, and solve it under various criteria. The ASIP model establishes bridges between statistical physics and queueing theory as it represents a tandem array of queueing systems with (unlimited) batch service, and a tandem array of growth-collapse processes.

  12. Inclusive fitness in agriculture

    PubMed Central

    Kiers, E. Toby; Denison, R. Ford

    2014-01-01

    Trade-offs between individual fitness and the collective performance of crop and below-ground symbiont communities are common in agriculture. Plant competitiveness for light and soil resources is key to individual fitness, but higher investments in stems and roots by a plant community to compete for those resources ultimately reduce crop yields. Similarly, rhizobia and mycorrhizal fungi may increase their individual fitness by diverting resources to their own reproduction, even if they could have benefited collectively by providing their shared crop host with more nitrogen and phosphorus, respectively. Past selection for inclusive fitness (benefits to others, weighted by their relatedness) is unlikely to have favoured community performance over individual fitness. The limited evidence for kin recognition in plants and microbes changes this conclusion only slightly. We therefore argue that there is still ample opportunity for human-imposed selection to improve cooperation among crop plants and their symbionts so that they use limited resources more efficiently. This evolutionarily informed approach will require a better understanding of how interactions among crops, and interactions with their symbionts, affected their inclusive fitness in the past and what that implies for current interactions. PMID:24686938

  13. Inclusive fitness in agriculture.

    PubMed

    Kiers, E Toby; Denison, R Ford

    2014-05-19

    Trade-offs between individual fitness and the collective performance of crop and below-ground symbiont communities are common in agriculture. Plant competitiveness for light and soil resources is key to individual fitness, but higher investments in stems and roots by a plant community to compete for those resources ultimately reduce crop yields. Similarly, rhizobia and mycorrhizal fungi may increase their individual fitness by diverting resources to their own reproduction, even if they could have benefited collectively by providing their shared crop host with more nitrogen and phosphorus, respectively. Past selection for inclusive fitness (benefits to others, weighted by their relatedness) is unlikely to have favoured community performance over individual fitness. The limited evidence for kin recognition in plants and microbes changes this conclusion only slightly. We therefore argue that there is still ample opportunity for human-imposed selection to improve cooperation among crop plants and their symbionts so that they use limited resources more efficiently. This evolutionarily informed approach will require a better understanding of how interactions among crops, and interactions with their symbionts, affected their inclusive fitness in the past and what that implies for current interactions. PMID:24686938

  14. Body Hair

    MedlinePlus

    ... 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, ...

  15. Bacterial differentiation.

    PubMed

    Shapiro, L; Agabian-Keshishian, N; Bendis, I

    1971-09-01

    technique can be used to select for mutants blocked in the various stages of morphogenesis. 3) Temperature-sensitive mutants of Caulobacter that are restricted in macromolecular synthesis and development at elevated temperatures have been isolated. 4) Genetic exchange in the Calflobacter genus has been demonstrated and is now being defined. Two questions related to control processes can now readily be approached experimentally. (i) Is the temporal progression of events occurring during bacterial differentiation controlled by regulator gene products? (ii) Is the differentiation cycle like a biosynthetic pathway where one event must follow another? The availability of temperature-sensitive mutants blocked at various stages of development permits access to both questions. An interesting feature of the differentiation cycle is that the polar organelle may represent a special segregated unit which is operative in the control of the differentiation process. Perhaps the sequential morphogenic changes exhibited by Caulobacter are dependent on the initial synthesis of this organelle. Because the ultimate expression of cell changes are dependent on selective protein synthesis, specific messenger RNA production-either from DNA present in an organelle or from the chromosome-may prove to be a controlling factor in cell differentiation. We have begun studies with RNA polymerase purified from Caulobacter crescentus to determine whether cell factors or alterations in the enzyme structure serve to change the specificity of transcription during the cell cycle. Control of sequential cell changes at the level of transcription has long been postulated and has recently been substantiated in the case of Bacillus sporulation (6). The Caulobacter bacteria now present another system in which direct analysis of these control mechanisms is feasible. PMID:5572165

  16. Inclusive Jets in PHP

    NASA Astrophysics Data System (ADS)

    Roloff, P.

    Differential inclusive-jet cross sections have been measured in photoproduction for boson virtualities Q^2 < 1 GeV^2 with the ZEUS detector at HERA using an integrated luminosity of 300 pb^-1. Jets were identified in the laboratory frame using the k_T, anti-k_T or SIScone jet algorithms. Cross sections are presented as functions of the jet pseudorapidity, eta(jet), and the jet transverse energy, E_T(jet). Next-to-leading-order QCD calculations give a good description of the measurements, except for jets with low E_T(jet) and high eta(jet). The cross sections have the potential to improve the determination of the PDFs in future QCD fits. Values of alpha_s(M_Z) have been extracted from the measurements based on different jet algorithms. In addition, the energy-scale dependence of the strong coupling was determined.

  17. The body and physiotherapy.

    PubMed

    Nicholls, David A; Gibson, Barbara E

    2010-11-01

    In recent years, physiotherapists have been increasingly interested in defining their professional identity. At the heart of this interest lies a fundamental question about the role that the body plays in defining physiotherapy practice. Given the importance of the body to physiotherapy, it is surprising how under-theorized the body is in existing physiotherapy literature. With a few notable exceptions, the body as a philosophical/theoretical construct has been almost entirely bypassed by the profession. In this paper the authors argue that a renewed interest in the meaning given to the body by physiotherapists is timely, and offer a sociohistorical critique of the role the body has played in defining physiotherapy practice. We challenge physiotherapists' longstanding affinity with a biomechanical view of the body, arguing that whilst this approach may have been critically important in the past, it is now increasingly clear that a more diverse and inclusive approach to the body will be needed in the future. The authors explore the notion of embodiment and suggest ways in which embodiment theory might be applied to physiotherapy practice. PMID:20795873

  18. Possibilities for an Inclusive Society in Singapore: Becoming Inclusive within

    ERIC Educational Resources Information Center

    Lim, Levan

    2009-01-01

    The envisioning of Singapore as an inclusive society has witnessed the most progressive systemic and policy developments concerning people with disabilities in recent years. The building of "heartware" in society (as in the will, values, and attitudes of its citizens) in order to realize the vision of an inclusive society, however, requires both…

  19. Student Teachers' Attitudes and Beliefs about Inclusion and Inclusive Practice

    ERIC Educational Resources Information Center

    Beacham, Nigel; Rouse, Martyn

    2012-01-01

    The beliefs and attitudes of teachers are an important element in the development of inclusive education and its associated practices. Teacher education is seen as crucial in helping to develop positive attitudes and beliefs that are thought to promote inclusion, although attempts to reform teacher education in order to address issues of inclusion…

  20. Inclusive Education: Identifying Teachers' Perceived Stressors in Inclusive Classrooms

    ERIC Educational Resources Information Center

    Brackenreed, Darlene

    2008-01-01

    This research replicates the study conducted by Forlin (2001) in Churchlands, Western Australia. Forlin's Inclusive Education Teacher Stress and Coping Questionnaire was adapted from the original questionnaire to more accurately reflect the language and practice of inclusion in Ontario (Frost & Brackenreed, 2004). The purpose of this study was to…

  1. Social Inclusion and Metrolingual Practices

    ERIC Educational Resources Information Center

    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…

  2. Serbian Teachers' Attitudes towards Inclusion

    ERIC Educational Resources Information Center

    Kalyva, Efrosini; Gojkovic, Dina; Tsakiris, Vlastaris

    2007-01-01

    This study investigated the attitudes of 72 Serbian teachers towards the inclusion of children with Special Educational Needs (SEN) in mainstream schools; they were asked to complete "My Thinking About Inclusion Questionnaire" (Stoiber, Goettinger, & Goetz, 1998). It was found that Serbian teachers held overall slightly negative attitudes towards…

  3. Principals Influence Culture of Inclusion.

    ERIC Educational Resources Information Center

    King, Sophia

    2000-01-01

    Three New American High Schools (in Pittsburgh, Baltimore, and Greenbelt, Maryland) have successfully replaced the special-education culture of separation with a culture of inclusion. A large part of moving from self-contained practices to inclusion was helping students become self-advocates, so that the world outside school and home would not…

  4. In Support of Unfinished Inclusion

    ERIC Educational Resources Information Center

    Hausstätter, Rune Sarromaa

    2014-01-01

    This article claims that the radical potential inherent in the origins of inclusive education has been altered into a tool for protecting the status quo. Drawing on ideas from the essay "The Unfinished" by Thomas Mathiesen (1971), I discuss inclusion as a potential alternative to mainstream education and argue that the potential power of…

  5. Building Inclusive Cities and Communities

    ERIC Educational Resources Information Center

    Freiler, Christa

    2008-01-01

    Canada prides itself on being an inclusive country. Immigrants from all over the world arrive in Canada's cities with their families because they feel welcome and safe. According to research, engagement towards social inclusion increased among Canadians during the last 30 last years. These changing values resulted in the creation of official…

  6. Friendship in Inclusive Physical Education

    ERIC Educational Resources Information Center

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

    2009-01-01

    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…

  7. Stiffening solids with liquid inclusions

    NASA Astrophysics Data System (ADS)

    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.

  8. Promoting a Lifetime of Inclusion.

    ERIC Educational Resources Information Center

    Renzaglia, Adelle; Karvonen, Meagan; Drasgow, Erik; Stoxen, Craig C.

    2003-01-01

    Discussion of ways to promote inclusion for individuals with severe disabilities first establishes the principle of normalization, then describes conditions and practices that reflect this principle and foster inclusion across the life span. These include universal design, person-centered planning, self-determination, and positive behavior…

  9. Early Childhood Inclusion in Turkey

    ERIC Educational Resources Information Center

    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…

  10. Inclusion in Malaysian Integrated Preschools

    ERIC Educational Resources Information Center

    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…

  11. Fluid Inclusions in Carbonaceous Chondrites

    NASA Technical Reports Server (NTRS)

    Saylor, J.; Zolensky, M. E.; Bodnar, R. J.; Le L.; Schwandt, C.

    2001-01-01

    Fluid inclusions are present in carbonaceous chondrites. Of the chondrites studied (CI1, CM1 and 2, CV3) fluid inclusions were found only in CM2s and CI1s, and by extrapolation are most likely to be found there in the future. Additional information is contained in the original extended abstract.

  12. Quality in Inclusive Preschool Classrooms

    ERIC Educational Resources Information Center

    Hestenes, Linda L.; Cassidy, Deborah J.; Shim, Jonghee; Hegde, Archana V.

    2008-01-01

    Research Findings: Quality of care for preschool children in inclusive and noninclusive classrooms was examined in two studies. In Study 1, comparisons across a large sample of classrooms (N = 1, 313) showed that inclusive classrooms were higher than noninclusive classrooms in global quality as well as on two dimensions of quality…

  13. Can VET Help Create a More Inclusive Society?

    ERIC Educational Resources Information Center

    Buddelmeyer, Hielke; Polidano, Cain

    2016-01-01

    This publication provides a summary of a program of research undertaken for the National Centre for Vocational Education Research (NCVER) by the Melbourne Institute of Applied Economic and Social Research between 2011 and 2014. Comprising six projects, the body of work focuses on the impact of education and training on social inclusion and on the…

  14. Inclusion Kinetics of Polyrotaxanes

    NASA Astrophysics Data System (ADS)

    Yokoyama, Hideaki; Takahashi, Shoko; Ito, Kohzo; Yamada, Norifumi

    Inclusion complex (IC) formation of α-cyclodextrin (α-CD) and poly(ethylene glycol) (PEG) brush in water was investigated by Surface Plasmon Resonance Spectroscopy(SPR), neutron reflectometry(NR) and grazing incident wide angle X-ray scattering(GISANS). Spontaneous IC formation of α-CD with PEG (polyrotaxanes) is believed to be due to hydrophobic interaction between the hydrophobic interior of α-CD and PEG; however, the detail of the IC formation kinetics has not been observed because IC formation results in aggregation and precipitation of the complex. SPR revealed that IC formation occurs after induction period, which often appears in crystallization. When concentration of α-CD solution is 10%, IC consisting randomly oriented α-CD polycrystal appeared. In contrast, when the concentration of α-CD solution is 5%, a uniform 10-nm-thick IC layer with α-CD stacked perpendicular to the substrate appeared. 10-nm-thick IC was also found in the diluted PEG brush in contact with a 10% α-CD solution. The characteristic 10-nm-thick layer is related to the folded crystalline structure of α-CD on PEG brush. Such crystallization was proved to be the main driving force for IC formation.

  15. Inclusion body disease in a great horned owl.

    PubMed

    Sileo, L; Carlson, H C; Crumley, S C

    1975-01-01

    The carcass of a great horned owl (Bubo virginianus), which had been found moribund in southern Ontario, was presented for necropsy. Throughout the liver and spleen were numerous white foci 1-2 mm in diameter; also noted were white plaques in the mucosae of the pharyngeal papillae and intestine. Results of light and electron microscopic studies and experimental transmission to two captive great horned owls suggested that this was a herpvirus disease similar and possibly indentical to the owl disease reported by other workers in Wiconsin and Australia. PMID:163384

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

    MedlinePlus

    ... called neuroserpin, which is found in nerve cells ( neurons ). Neuroserpin plays a role in the development and ... system. This protein helps control the growth of neurons and their connections with one another, which suggests ...

  17. The infidelity of melt inclusions?

    NASA Astrophysics Data System (ADS)

    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

  18. A Deadly Path: Bacterial Spread During Bubonic Plague.

    PubMed

    Gonzalez, Rodrigo J; Miller, Virginia L

    2016-04-01

    Yersinia pestis causes bubonic plague, a fulminant disease where host immune responses are abrogated. Recently developed in vivo models of plague have resulted in new ideas regarding bacterial spread in the body. Deciphering bacterial spread is key to understanding Y. pestis and the immune responses it encounters during infection. PMID:26875618

  19. Bacterial adenosine triphosphate as a measure of urinary tract infection

    NASA Technical Reports Server (NTRS)

    Chappelle, E. W.; Picciolo, G. L.

    1971-01-01

    Procedure detects and counts bacteria present in urine samples. Method also determines bacterial levels in other aqueous body fluids including lymph fluid, plasma, blood, spinal fluid, saliva and mucous.

  20. Nontraditional Student Perceptions of Collegiate Inclusion

    ERIC Educational Resources Information Center

    Witkowsky, Patricia; Mendez, Sylvia; Ogunbowo, Oluwafolakemi; Clayton, Grant; Hernandez, Nancy

    2016-01-01

    This study explored student responses to a Student Inclusiveness Survey (SIS), with specific attention to nontraditional student responses about collegiate inclusion. Specifically, the SIS constructs that related to inclusion, the Perceptions of Inclusiveness and Institutional Safeguarding of Inclusiveness, were analyzed descriptively, and…

  1. The impact of bacterial and viral co‐infection in severe influenza

    PubMed Central

    Blyth, Christopher C.; Webb, Steve A. R.; Kok, Jen; Dwyer, Dominic E.; van Hal, Sebastiaan J.; Foo, Hong; Ginn, Andrew N.; Kesson, Alison M.; Seppelt, Ian; Iredell, Jonathan R.

    2013-01-01

    Please cite this paper as: Blyth et al. (2013) The impact of bacterial and viral co‐infection in severe influenza. Influenza and Other Respiratory Viruses 7(2) 168–176. Background  Many questions remain concerning the burden, risk factors and impact of bacterial and viral co‐infection in patients with pandemic influenza admitted to the intensive care unit (ICU). Objectives  To examine the burden, risk factors and impact of bacterial and viral co‐infection in Australian patients with severe influenza. Patients/Methods  A cohort study conducted in 14 ICUs was performed. Patients with proven influenza A during the 2009 influenza season were eligible for inclusion. Demographics, risk factors, clinical data, microbiological data, complications and outcomes were collected. Polymerase chain reaction for additional bacterial and viral respiratory pathogens was performed on stored respiratory samples. Results  Co‐infection was identified in 23·3–26·9% of patients with severe influenza A infection: viral co‐infection, 3·2–3·4% and bacterial co‐infection, 20·5–24·7%. Staphylococcus aureus was the most frequent bacterial co‐infection followed by Streptococcus pneumoniae and Haemophilus influenzae. Patients with co‐infection were younger [mean difference in age = 8·46 years (95% CI: 0·18–16·74 years)], less likely to have significant co‐morbidities (32·0% versus 66·2%, P = 0·004) and less frequently obese [mean difference in body mass index = 6·86 (95% CI: 1·77–11·96)] compared to those without co‐infection. Conclusions  Bacterial or viral co‐infection complicated one in four patients admitted to ICU with severe influenza A infection. Despite the co‐infected patients being younger and with fewer co‐morbidities, no significant difference in outcomes was observed. It is likely that co‐infection contributed to a need for ICU admission in those without other risk factors for severe influenza disease

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

    SciTech Connect

    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

  3. Electromagnetism of Bacterial Growth

    NASA Astrophysics Data System (ADS)

    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.

  4. STIM1 Is a Novel Component of ER-Chlamydia trachomatis Inclusion Membrane Contact Sites

    PubMed Central

    Agaisse, Hervé; Derré, Isabelle

    2015-01-01

    Productive developmental cycle of the obligate intracellular bacterial pathogen Chlamydia trachomatis depends on the interaction of the replicative vacuole, named the inclusion, with cellular organelles. We have recently reported the formation of ER-Inclusion membrane contact sites (MCSs), where the endoplasmic reticulum (ER) is in apposition to the inclusion membrane. These platforms contain the C. trachomatis inclusion membrane protein IncD, the mammalian ceramide transfer protein CERT and the ER resident proteins VAPA/B and were proposed to play a role in the non-vesicular trafficking of lipids to the inclusion. Here, we identify STIM1 as a novel component of ER-Inclusion MCSs. STIM1, an ER calcium (Ca2+) sensor that relocate to ER-Plasma Membrane (PM) MCSs upon Ca2+ store depletion, associated with C. trachomatis inclusion. STIM1, but not the general ER markers Rtn3C and Sec61ß, was enriched at the inclusion membrane. Ultra-structural studies demonstrated that STIM1 localized to ER-Inclusion MCSs. Time-course experiments showed that STIM1, CERT and VAPB co-localized throughout the developmental cycle. By contrast, Orai1, the PM Ca2+ channel that interacts with STIM1 at ER-PM MCSs, did not associate with C. trachomatis inclusion. Upon ER Ca2+ store depletion, a pool of STIM1 relocated to ER-PM MCSs, while the existing ER-Inclusion MCSs remained enriched in STIM1. Finally, we have identified the CAD domain, which mediates STIM1-Orai1 interaction, as the minimal domain required for STIM1 enrichment at ER-Inclusion MCSs. Altogether this study identifies STIM1 as a novel component of ER-C. trachomatis inclusion MCSs. We discuss the potential role(s) of STIM1 during the infection process. PMID:25915399

  5. CT -- Body

    MedlinePlus

    ... News Physician Resources Professions Site Index A-Z Computed Tomography (CT) - Body Computed tomography (CT) of the body uses special x-ray ... Body? What is CT Scanning of the Body? Computed tomography, more commonly known as a CT or CAT ...

  6. Chemical behaviour of metallic inclusions in glass melts

    NASA Astrophysics Data System (ADS)

    Stachel, Dörte; Zangenberg, Frauke; Müller, Thomas E.

    2007-05-01

    Inclusions of all kinds cannot be accepted in glass articles. Most of them are non-metallic stones, silicon spheres, cords, bubbles, and sometimes metallic inclusions. Identification of every appearance is not possible, since body structure and chemical composition do not compare. Laboratory experiments regarding the behaviour of the metals copper, brass and iron and their dissolution mechanisms in glass melts at technological relevant temperatures were analysed. Besides partial oxidation processes, sulphidic species were found. The experiments have been compared with thermodynamic calculations of several reaction mechanisms.

  7. Inclusion of Fermented Foods in Food Guides around the World

    PubMed Central

    Chilton, Stephanie N.; Burton, Jeremy P.; Reid, Gregor

    2015-01-01

    Fermented foods have been a well-established part of the human diet for thousands of years, without much of an appreciation for, or an understanding of, their underlying microbial functionality, until recently. The use of many organisms derived from these foods, and their applications in probiotics, have further illustrated their impact on gastrointestinal wellbeing and diseases affecting other sites in the body. However, despite the many benefits of fermented foods, their recommended consumption has not been widely translated to global inclusion in food guides. Here, we present the case for such inclusion, and challenge health authorities around the world to consider advocating for the many benefits of these foods. PMID:25580813

  8. Tidal disruption of solid bodies

    NASA Technical Reports Server (NTRS)

    Dobrovolskis, Anthony R.

    1990-01-01

    The problem of stress, strain, and breakup in solid satellites and stray bodies subject to tidal perturbations is presently addressed in view of three novel considerations. After presenting a new analytic solution for the stress tensor in a homogeneous and compressible elastic sphere, where the inclusion of compressibility alters stresses by several percent, realistic failure criteria are noted to demonstrate the general failure of such ductile bodies as iron meteoroids by plastic shear, while brittle ice bodies fail by either tensile or shear fracture. A reexamination of crack propagation after initial failure allows the diverse breakup criteria to be reconciled.

  9. Innovation for an Inclusive Future

    NASA Astrophysics Data System (ADS)

    Springett, Mark; Rice, Mark; Carmichael, Alex; Griffiths, Richard

    This workshop will focus on setting the agenda for research, practice and policy in support of inclusive design for third generation computer-based products. The next generation of technology represents an unprecedented opportunity to improve the quality of life for groups of users who have previously faced exclusion, such as those with impairments and older citizens. At the same time it risks creating a greater digital divide and further exclusion. How we approach design for this new generation will determine whether or not the third wave will provide positive advances towards an inclusive digital world. We therefore need to put forward both a rationale for inclusive design and provide pointers towards technical development and design practice in support of inclusion. It is our belief that there is not only a strong moral case for design for inclusion but also significant commercial incentive, which may be key to persuading influential players to focus on inclusion. Therefore one of our key objectives is to describe and promote the advantages of designing ‘in from the edges’ of the user population rather than designing for a notional ‘average’ user.

  10. Phylogenetic mapping of bacterial morphology

    NASA Technical Reports Server (NTRS)

    Siefert, J. L.; Fox, G. E.

    1998-01-01

    The availability of a meaningful molecular phylogeny for bacteria provides a context for examining the historical significance of various developments in bacterial evolution. Herein, the classical morphological descriptions of selected members of the domain Bacteria are mapped upon the genealogical ancestry deduced from comparison of small-subunit rRNA sequences. For the species examined in this study, a distinct pattern emerges which indicates that the coccus shape has arisen and accumulated independently multiple times in separate lineages and typically survived as a persistent end-state morphology. At least two other morphologies persist but have evolved only once. This study demonstrates that although bacterial morphology is not useful in defining bacterial phylogeny, it is remarkably consistent with that phylogeny once it is known. An examination of the experimental evidence available for morphogenesis as well as microbial fossil evidence corroborates these findings. It is proposed that the accumulation of persistent morphologies is a result of the biophysical properties of peptidoglycan and their genetic control, and that an evolved body-plan strategy based on peptidoglycan may have been a fate-sealing step in the evolution of Bacteria. More generally, this study illustrates that significant evolutionary insights can be obtained by examining biological and biochemical data in the context of a reliable phylogenetic structure.

  11. Bacterial infections complicating tongue piercing

    PubMed Central

    Yu, Catherine HY; Minnema, Brian J; Gold, Wayne L

    2010-01-01

    Tongue piercing has become an increasingly popular form of body art. However, this procedure can occasionally be complicated by serious bacterial infections. The present article reports a case of prosthetic valve endocarditis caused by a Gemella species in a patient with a pierced tongue, and reviews 18 additional cases of local and systemic bacterial infections associated with tongue piercing. Infections localized to the oral cavity and head and neck region included molar abscess, glossal abscess, glossitis, submandibular lymphadenitis, submandibular sialadenitis, Ludwig’s angina and cephalic tetanus. Infections distal to the piercing site included eight cases of infective endocarditis, one case of chorioamnionitis and one case of cerebellar abscess. Oropharyngeal flora were isolated from all cases. While bacterial infections following tongue piercing are rare, there are reports of potentially life-threatening infections associated with the procedure. Both piercers and their clients should be aware of these potential complications, and standardized infection prevention and control practices should be adopted by piercers to reduce the risk. PMID:21358880

  12. Facial bacterial infections: folliculitis.

    PubMed

    Laureano, Ana Cristina; Schwartz, Robert A; Cohen, Philip J

    2014-01-01

    Facial bacterial infections are most commonly caused by infections of the hair follicles. Wherever pilosebaceous units are found folliculitis can occur, with the most frequent bacterial culprit being Staphylococcus aureus. We review different origins of facial folliculitis, distinguishing bacterial forms from other infectious and non-infectious mimickers. We distinguish folliculitis from pseudofolliculitis and perifolliculitis. Clinical features, etiology, pathology, and management options are also discussed. PMID:25441463

  13. Demonstrating Bacterial Flagella.

    ERIC Educational Resources Information Center

    Porter, John R.; And Others

    1992-01-01

    Describes an effective laboratory method for demonstrating bacterial flagella that utilizes the Proteus mirabilis organism and a special harvesting technique. Includes safety considerations for the laboratory exercise. (MDH)

  14. Small molecule control of bacterial biofilms

    PubMed Central

    Worthington, Roberta J.; Richards, Justin J.

    2012-01-01

    Bacterial biofilms are defined as a surface attached community of bacteria embedded in a matrix of extracellular polymeric substances that they have produced. When in the biofilm state, bacteria are more resistant to antibiotics and the host immune response than are their planktonic counterparts. Biofilms are increasingly recognized as being significant in human disease, accounting for 80% of bacterial infections in the body and diseases associated with bacterial biofilms include: lung infections of cystic fibrosis, colitis, urethritis, conjunctivitis, otitis, endocarditis and periodontitis. Additionally, biofilm infections of indwelling medical devices are of particular concern, as once the device is colonized infection is virtually impossible to eradicate. Given the prominence of biofilms in infectious diseases, there has been an increased effort toward the development of small molecules that will modulate bacterial biofilm development and maintenance. In this review, we highlight the development of small molecules that inhibit and/or disperse bacterial biofilms through non-microbicidal mechanisms. The review discuses the numerous approaches that have been applied to the discovery of lead small molecules that mediate biofilm development. These approaches are grouped into: 1) the identification and development of small molecules that target one of the bacterial signaling pathways involved in biofilm regulation, 2) chemical library screening for compounds with anti-biofilm activity, and 3) the identification of natural products that possess anti-biofilm activity, and the chemical manipulation of these natural products to obtain analogues with increased activity. PMID:22733439

  15. Body Measurement.

    ERIC Educational Resources Information Center

    Neufeld, K. Allen

    1989-01-01

    Described are activities for measuring the human body. The activities include measurements and calculations, calculating volume and density, problems related to body measurement, and using a nomogram. Several charts, illustrations, and a nomogram are provided. (YP)

  16. Experimental Simulation of Shock Reequilibration of Fluid Inclusions During Meteorite Impact

    NASA Technical Reports Server (NTRS)

    Madden, M. E. Elwood; Hoerz, R. J.; Bodnar, R. J.

    2003-01-01

    Fluid inclusions are microscopic volumes of fluid trapped within minerals as they precipitate. Fluid inclusions are common in terrestrial minerals formed under a wide array of geological settings from surface evaporite deposits to kimberlite pipes. While fluid inclusions in terrestrial rocks are the rule rather than the exception, only few fluid inclusion-bearing meteorites have been documented. The rarity of fluid inclusions in meteoritic material may be explained in two ways. First, it may reflect the absence of fluids (water?) on meteorite parent bodies. Alternatively, fluids may have been present when the rock formed, but any fluid inclusions originally trapped on the parent body were destroyed by the extreme P-T conditions meteorites often experience during impact events. Distinguishing between these two possibilities can provide significant constraints on the likelihood of life on the parent body. Just as textures, structures, and compositions of mineral phases can be significantly altered by shock metamorphism upon hypervelocity impact, fluid inclusions contained within component minerals may be altered or destroyed due to the high pressures, temperatures, and strain rates associated with impact events. Reequilibration may occur when external pressure-temperature conditions differ significantly from internal fluid isochoric conditions, and result in changes in fluid inclusion properties and/or textures. Shock metamorphism and fluid inclusion reequilibration can affect both the impacted target material and the meteoritic projectile. By examining the effects of shock deformation on fluid inclusion properties and textures we may be able to better constrain the pressure-temperature path experienced by shocked materials and also gain a clearer understanding of why fluid inclusions are rarely found in meteoritic samples.

  17. Ancient asteroids enriched in refractory inclusions.

    PubMed

    Sunshine, J M; Connolly, H C; McCoy, T J; Bus, S J; La Croix, L M

    2008-04-25

    Calcium- and aluminum-rich inclusions (CAIs) occur in all classes of chondritic meteorites and contain refractory minerals predicted to be the first condensates from the solar nebula. Near-infrared spectra of CAIs have strong 2-micrometer absorptions, attributed to iron oxide-bearing aluminous spinel. Similar absorptions are present in the telescopic spectra of several asteroids; modeling indicates that these contain approximately 30 +/- 10% CAIs (two to three times that of any meteorite). Survival of these undifferentiated, large (50- to 100-kilometer diameter) CAI-rich bodies suggests that they may have formed before the injection of radiogenic 26Al into the solar system. They have also experienced only modest post-accretionary alteration. Thus, these asteroids have higher concentrations of CAI material, appear less altered, and are more ancient than any known sample in our meteorite collection, making them prime candidates for sample return. PMID:18356491

  18. Diverse Perspectives on Inclusive School Communities

    ERIC Educational Resources Information Center

    Tsokova, Diana; Tarr, Jane

    2012-01-01

    What is an inclusive school community? How do stakeholders perceive their roles and responsibilities towards inclusive school communities? How can school communities become more inclusive through engagement with individual perspectives? "Diverse Perspectives on Inclusive School Communities" captures and presents the voices of a wide range of…

  19. Interpreting Inclusivity: An Endeavour of Great Proportions

    ERIC Educational Resources Information Center

    Berlach, Richard George; Chambers, Dianne Joy

    2011-01-01

    Commencing with a historical account of how special needs education has informed the inclusivity debate, the authors consider the knotty problem of what is meant by inclusivity. An examination of the characteristics of inclusivity is then undertaken, and a functional school-based inclusivity framework--a three-faceted model--is proposed. The model…

  20. Vimentin in Bacterial Infections

    PubMed Central

    Mak, Tim N.; Brüggemann, Holger

    2016-01-01

    Despite well-studied bacterial strategies to target actin to subvert the host cell cytoskeleton, thus promoting bacterial survival, replication, and dissemination, relatively little is known about the bacterial interaction with other components of the host cell cytoskeleton, including intermediate 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 virulence factors that target vimentin to subvert its function in order to change the host cell fate in the course of a bacterial infection. PMID:27096872

  1. Bacterial Ion Channels.

    PubMed

    Compton, Emma L R; Mindell, Joseph A

    2010-09-01

    Bacterial ion channels were known, but only in special cases, such as outer membrane porins in Escherichia coli and bacterial toxins that form pores in their target (bacterial or mammalian) membranes. The exhaustive coverage provided by a decade of bacterial genome sequencing has revealed that ion channels are actually widespread in bacteria, with homologs of a broad range of mammalian channel proteins coded throughout the bacterial and archaeal kingdoms. This review discusses four groups of bacterial channels: porins, mechano-sensitive (MS) channels, channel-forming toxins, and bacterial homologs of mammalian channels. The outer membrane (OM) of gram-negative bacteria blocks access of essential nutrients; to survive, the cell needs to provide a mechanism for nutrients to penetrate the OM. Porin channels provide this access by forming large, nonspecific aqueous pores in the OM that allow ions and vital nutrients to cross it and enter the periplasm. MS channels act as emergency release valves, allowing solutes to rapidly exit the cytoplasm and to dissipate the large osmotic disparity between the internal and external environments. MS channels are remarkable in that they do this by responding to forces exerted by the membrane itself. Some bacteria produce toxic proteins that form pores in trans, attacking and killing other organisms by virtue of their pore formation. The review focuses on those bacterial toxins that kill other bacteria, specifically the class of proteins called colicins. Colicins reveal the dangers of channel formation in the plasma membrane, since they kill their targets with exactly that approach. PMID:26443789

  2. Vimentin in Bacterial Infections.

    PubMed

    Mak, Tim N; Brüggemann, Holger

    2016-01-01

    Despite well-studied bacterial strategies to target actin to subvert the host cell cytoskeleton, thus promoting bacterial survival, replication, and dissemination, relatively little is known about the bacterial interaction with other components of the host cell cytoskeleton, including intermediate 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 virulence factors that target vimentin to subvert its function in order to change the host cell fate in the course of a bacterial infection. PMID:27096872

  3. Fungal and Bacterial Diseases.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fungal and bacterial diseases are important constraints to production. Recognition of diseases and information on their biology is important in disease management. This chapter is aimed at providing diagnostic information on fungal and bacterial diseases of sugar beet and their biology, epidemiolo...

  4. Protein inclusions produced by the entomopathogenic bacterium Xenorhabdus nematophilus subsp. nematophilus.

    PubMed Central

    Couche, G A; Gregson, R P

    1987-01-01

    The entomopathogenic bacterium Xenorhabdus nematophilus subsp. nematophilus produces two types of intracellular inclusion bodies during in vitro culture. Large cigar-shaped inclusions (designated type 1) and smaller ovoid inclusions (designated type 2) were purified from cell lysates, using differential centrifugation in discontinuous glycerol gradients and isopycnic density gradient centrifugation in sodium diatrizoate. The inclusions, composed almost exclusively of protein, are readily soluble at high and low pH values and in the presence of cation chelators such as EDTA, anionic detergents (sodium dodecyl sulfate), or protein denaturants (urea, NaBr). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified inclusions revealed a single 26-kilodalton protein (IP-1) in type 1 inclusions and a 22-kilodalton protein (IP-2) in type 2 inclusions. Analysis of these proteins by isoelectric focusing in the presence of 8 M urea showed that IP-1 is acidic and IP-2 is neutral. Furthermore, each protein occurred in multiple forms differing slightly in isoelectric point. Other variations in peptides released by trypsin digestion, immunological properties, and amino acid composition revealed significant structural differences between IP-1 and IP-2. Kinetic studies using light microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblotting procedures showed that inclusion protein synthesis occurs only during the second half of exponential culture growth. Synthesis of inclusion proteins and their aggregation to form inclusions occurred concurrently. Possible functions for these abundant proteins are discussed. Images PMID:3667532

  5. Special Teaching for Special Children? Pedagogies for Inclusion. Inclusive Education

    ERIC Educational Resources Information Center

    Lewis, Ann, Ed.; Norwich, Brahm, Ed.

    2004-01-01

    Some special needs groups (for example dyslexia) have argued strongly for the need for particular specialist approaches. In contrast, many proponents of inclusion have argued that "good teaching is good teaching for all" and that all children benefit from similar approaches. Both positions fail to scrutinise this issue rigorously and coherently,…

  6. The Inclusion of Music/the Music of Inclusion

    ERIC Educational Resources Information Center

    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.…

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

    ERIC Educational Resources Information Center

    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…

  8. The Contours of Inclusion: Inclusive Arts Teaching and Learning

    ERIC Educational Resources Information Center

    Glass, Don; Henderson, Bill; Barnum, Leah; Kronenberg, Deborah; Blair, Kati; Jenkins, Richard; Hurel, Nicole Agois

    2010-01-01

    The purpose of this publication is to share models and case examples of the process of inclusive arts curriculum design and evaluation. The first section explains the conceptual and curriculum frameworks that were used in the analysis and generation of the featured case studies (i.e. Understanding by Design, Differentiated Instruction, and…

  9. The Needs of Inclusive Preschool Teachers about Inclusive Practices

    ERIC Educational Resources Information Center

    Akalin, Selma; Demir, Seyda; Sucuoglu, Bülbin; Bakkaloglu, Hatice; Iscen, Fadime

    2014-01-01

    Problem Statement: Since the law related to the inclusion of children with special needs was passed, the number of children with special needs in preschool classrooms has increased each year. Preschool teachers face serious challenges when they teach children with diverse abilities in the same environment. Although most of them support the idea of…

  10. Developing Inclusive Practice in Scotland: The National Framework for Inclusion

    ERIC Educational Resources Information Center

    Barrett, Louise; Beaton, Mhairi; Head, George; McAuliffe, Lisa; Moscardini, Lio; Spratt, Jennifer; Sutherland, Margaret

    2015-01-01

    This paper reports on the collaborative development of a "National Framework for Inclusion" under the auspices of the Scottish Teacher Education Committee by a working party representing each of the Scottish Universities providing initial teacher education. Recent research, international legislation and Scottish education policy have…

  11. Towards understanding microvillus inclusion disease.

    PubMed

    Vogel, Georg F; Hess, Michael W; Pfaller, Kristian; Huber, Lukas A; Janecke, Andreas R; Müller, Thomas

    2016-12-01

    Microvillus inclusion disease (MVID) is characterised by onset of intractable life-threatening watery diarrhoea during infancy. Transmission electron microscopy demonstrates shortening or absence of apical microvilli, pathognomonic microvillus inclusions in mature enterocytes and subapical accumulation of periodic acid-Schiff-positive granules or vesicles confirming diagnosis. Mutations in MYO5B have been found to cause MVID. In two patients with MVID, whole-exome sequencing of DNA revealed homozygous truncating mutations in STX3. Mutations in these genes disrupt trafficking between apical cargo vesicles and the apical plasma membrane. Thus, disturbed delivery of certain brush border membrane proteins is a common defect in MVID. PMID:26830108

  12. Bacterial carbon utilization in vertical subsurface flow constructed wetlands.

    PubMed

    Tietz, Alexandra; Langergraber, Günter; Watzinger, Andrea; Haberl, Raimund; Kirschner, Alexander K T

    2008-03-01

    Subsurface vertical flow constructed wetlands with intermittent loading are considered as state of the art and can comply with stringent effluent requirements. It is usually assumed that microbial activity in the filter body of constructed wetlands, responsible for the removal of carbon and nitrogen, relies mainly on bacterially mediated transformations. However, little quantitative information is available on the distribution of bacterial biomass and production in the "black-box" constructed wetland. The spatial distribution of bacterial carbon utilization, based on bacterial (14)C-leucine incorporation measurements, was investigated for the filter body of planted and unplanted indoor pilot-scale constructed wetlands, as well as for a planted outdoor constructed wetland. A simple mass-balance approach was applied to explain the bacterially catalysed organic matter degradation in this system by comparing estimated bacterial carbon utilization rates with simultaneously measured carbon reduction values. The pilot-scale constructed wetlands proved to be a suitable model system for investigating microbial carbon utilization in constructed wetlands. Under an ideal operating mode, the bulk of bacterial productivity occurred within the first 10cm of the filter body. Plants seemed to have no significant influence on productivity and biomass of bacteria, as well as on wastewater total organic carbon removal. PMID:17991505

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

    PubMed Central

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

    2010-01-01

    Summary 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 center (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. PMID:20331642

  14. Comment on "ancient asteroids enriched in refractory inclusions".

    PubMed

    Hezel, Dominik C; Russell, Sara S

    2008-11-14

    Sunshine et al. (Reports, 25 April 2008, p. 514) reported that certain asteroids contain 30 +/- 10 volume percent calcium- and aluminum-rich inclusions (CAIs). We contend that the amount of CAIs in CV chondrites is two to three times as low as the 10 volume percent assumed by the authors; thus, we question whether the CAI-rich bodies they studied are indeed older than known asteroids or formed before the injection of (26)Al into the solar nebula. PMID:19008430

  15. Classroom Management in Inclusive Settings

    ERIC Educational Resources Information Center

    Soodak, Leslie C.

    2003-01-01

    The inclusion of children with disabilities in general education classes provides an opportunity for teachers to identify classroom management policies and practices that promote diversity and community. Community-building management strategies that facilitate friendships, collaboration, parent involvement, and address challenging behaviors in a…

  16. The Inclusive Education in Europe

    ERIC Educational Resources Information Center

    Manzano-García, Beatriz; Fernández, María Tomé

    2016-01-01

    One of the phenomena that is of most concern to educational policy in Europe is immigration due to the fact that this is the source of new educational needs. This research looks at how European educational legislation deals with this topic. For this intercultural values that make inclusive education will be evaluated, we will analyze intercultural…

  17. Managing Special and Inclusive Education

    ERIC Educational Resources Information Center

    Rayner, Steve G.

    2007-01-01

    This book is a guide to special and inclusive education and provides a comprehensive overview of this complex field. Author Stephen Rayner examines context, policy, and practice, and shows how to successfully navigate the managerial challenges involved, while contributing to the way forward through leadership in a diverse field. The author…

  18. Grasping the Promise of Inclusion.

    ERIC Educational Resources Information Center

    Rudd, Fern

    A teacher and mother of a child with mental retardation examined the history and current status of the inclusion movement. A review of the historical background considers the origins of special education, the Education for All Handicapped Children Act of 1975, major court cases, and requirements of the 1997 amendments to the Individuals with…

  19. Inclusion in the Microsoft Workforce

    ERIC Educational Resources Information Center

    Exceptional Parent, 2008

    2008-01-01

    Since 1975, Microsoft has been a worldwide leader in software, services, and solutions that help people and businesses realize their full potential. Loren Mikola, the Disability Inclusion Program Manager at Microsoft, ensures that this technology also reaches and includes the special needs population and, through the hiring of individuals with…

  20. Early Childhood Inclusion in Croatia

    ERIC Educational Resources Information Center

    Ljubešic, Marta; Šimleša, Sanja

    2016-01-01

    This article explains early childhood inclusion in Croatia from its beginnings up to challenges in current policy and practice. The first preschool education for children with disabilities dates back to the 1980s and was provided in special institutions. In the last 10 years, mainstream kindergartens have been enrolling children with disabilities…

  1. Supporting General Educators' Inclusive Practices.

    ERIC Educational Resources Information Center

    Coombs-Richardson, Rita; Mead, Jean

    2001-01-01

    This article describes Project Inclusion, a state-funded project at Southeastern Louisiana University, which provided financial support for general educators to take university courses to develop their knowledge and skills concerning students with disabilities. The courses emphasized collaboration techniques, curricular modifications, and behavior…

  2. Taking Inclusion into the Future.

    ERIC Educational Resources Information Center

    Lipsky, Dorothy Kerzner; Gartner, Alan

    1998-01-01

    According to the reauthorized Individuals with Disabilities Education Act (1997), education of disabled children should produce outcomes akin to those expected of "regular" students, and disabled students should be educated with other kids. Implementing inclusive programs will require visionary leadership, educator collaboration, refocused…

  3. Tracing Inclusion: Determining Teacher Attitudes

    ERIC Educational Resources Information Center

    Logan, Brenda E.; Wimer, Gregory

    2013-01-01

    Though there appears to be an onslaught of No Child Left Behind, there is still more emphasis on testing than ever before. With the new implementation of national common-core standards, many school districts have moved towards full inclusive classrooms. However, it is rare that teachers have any input on whether such major decisions are apropos…

  4. Inclusive Education and the Arts

    ERIC Educational Resources Information Center

    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…

  5. Early Childhood Inclusion in Israel

    ERIC Educational Resources Information Center

    Al-Yagon, Michal; Aram, Dorit; Margalit, Malka

    2016-01-01

    This article describes conceptual aspects, current policies and practices, and research representing the Israeli perspective regarding early childhood inclusion (ECI) at preschool ages (3-6 years). We review legislative, historical, attitudinal, philosophical, practical, empirical, and cultural issues regarding ECI in Israel. Finally, we focus on…

  6. Curriculum Adaptation for Inclusive Classrooms.

    ERIC Educational Resources Information Center

    Neary, Tom; And Others

    This manual on curriculum adaptation for inclusive classrooms was developed as part of the PEERS (Providing Education for Everyone in Regular Schools) Project, a 5-year collaborative systems change project in California to facilitate the integration of students with severe disabilities previously at special centers into services at regular school…

  7. Towards Inclusion: An Australian Perspective

    ERIC Educational Resources Information Center

    Forbes, Fiona

    2007-01-01

    This article outlines the views of the Australian Special Education Principals' Association (ASEPA) on inclusion and the impact this is having on Australian Government Schools from a school based perspective. ASEPA is a relatively young association and was formed in 1997 out of the need to put forward the case to support students with special…

  8. Inclusive Education: Programmes and Provisions

    ERIC Educational Resources Information Center

    Kappen, Mini Dejo

    2010-01-01

    Inclusive education is a practice of teaching handicapped children in regular classrooms with non-handicapped children to the fullest extent possible; such children may have orthopedic, intellectual, emotional, or visual difficulties or handicaps associated with hearing or learning. In India there are constitutional provisions for Inclusive…

  9. Developing Movement as Inclusive Pedagogy

    ERIC Educational Resources Information Center

    Peter, Melanie; Walter, Ofra

    2010-01-01

    This article details the emergence of a training framework to support professional development in inclusive Movement teaching. This arose from a collaborative research project in spring 2008 (supported by the Training and Development Agency, UK), between two universities in England and Israel. Movement education is surprisingly underused globally,…

  10. Early Childhood Inclusion in Australia

    ERIC Educational Resources Information Center

    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…

  11. ABC transporters: bacterial exporters.

    PubMed Central

    Fath, M J; Kolter, R

    1993-01-01

    The ABC transporters (also called traffic ATPases) make up a large superfamily of proteins which share a common function and a common ATP-binding domain. ABC transporters are classified into three major groups: bacterial importers (the periplasmic permeases), eukaryotic transporters, and bacterial exporters. We present a comprehensive review of the bacterial ABC exporter group, which currently includes over 40 systems. The bacterial ABC exporter systems are functionally subdivided on the basis of the type of substrate that each translocates. We describe three main groups: protein exporters, peptide exporters, and systems that transport nonprotein substrates. Prototype exporters from each group are described in detail to illustrate our current understanding of this protein family. The prototype systems include the alpha-hemolysin, colicin V, and capsular polysaccharide exporters from Escherichia coli, the protease exporter from Erwinia chrysanthemi, and the glucan exporters from Agrobacterium tumefaciens and Rhizobium meliloti. Phylogenetic analysis of the ATP-binding domains from 29 bacterial ABC exporters indicates that the bacterial ABC exporters can be divided into two primary branches. One branch contains the transport systems where the ATP-binding domain and the membrane-spanning domain are present on the same polypeptide, and the other branch contains the systems where these domains are found on separate polypeptides. Differences in substrate specificity do not correlate with evolutionary relatedness. A complete survey of the known and putative bacterial ABC exporters is included at the end of the review. PMID:8302219

  12. A Petrologic Study of the IAB Iron Meteorites: Constraints on the Formation of the IAB-Winonaite Parent Body

    NASA Technical Reports Server (NTRS)

    Benedix, G. K.; McCoy, T. J.; Keil, K.; Love, S. G.

    1998-01-01

    We have studied IAB iron meteorites and their silicate-bearing inclusions to elucidate the origin of their parent body. We have divided IAB irons into five categories which best describe the inclusions and other properties of the irons.

  13. Primordial Compositions of Refractory Inclusions

    SciTech Connect

    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.

  14. Body Piercing

    PubMed Central

    Koenig, Laura M; Carnes, Molly

    1999-01-01

    OBJECTIVE To review the current information on medical complications, psychological implications, and legislative issues related to body piercing, a largely unregulated industry in the United States. METHODS We conducted a MEDLINE search of English language articles from 1966 until May 1998 using the search terms “body piercing” and “ear piercing.” Bibliographies of these references were reviewed for additional citations. We also conducted an Internet search for “body piercing” on the World Wide Web. MAIN RESULTS: In this manuscript, we review the available body piercing literature. We conclude that body piercing is an increasingly common practice in the United States, that this practice carries substantial risk of morbidity, and that most body piercing in the United States is being performed by unlicensed, unregulated individuals. Primary care physicians are seeing growing numbers of patients with body pierces. Practitioners must be able to recognize, treat, and counsel patients on body piercing complications and be alert to associated psychological conditions in patients who undergo body piercing. PMID:10354260

  15. High yield soluble bacterial expression and streamlined purification of recombinant human interferon α-2a.

    PubMed

    Bis, Regina L; Stauffer, Tara M; Singh, Surinder M; Lavoie, Thomas B; Mallela, Krishna M G

    2014-07-01

    Interferon α-2a (IFNA2) is a member of the Type I interferon cytokine family, known for its antiviral and anti-proliferative functions. The role of this family in the innate immune response makes it an attractive candidate for the treatment of many viral and chronic immune-compromised diseases. Recombinant IFNA2 is clinically used to modulate hairy cell leukemia as well as hepatitis c. Historically, IFNA2 has been purified from human leukocytes as well as bacterial expression systems. In most cases, bacterial expression of IFNA2 resulted in inclusion body formation, or required numerous purification steps that decreased the protein yield. Here, we describe an expression and purification scheme for IFNA2 using a pET-SUMO bacterial expression system and a single purification step. Using the SUMO protein as the fusion tag achieved high soluble protein expression. The SUMO tag was cleaved with the Ulp1 protease leaving no additional amino acids on the fusion terminus following cleavage. Mass spectrometry, circular dichroism, 2D heteronuclear NMR, and analytical ultracentrifugation confirmed the amino acid sequence identity, secondary and tertiary protein structures, and the solution behavior of the purified IFNA2. The purified protein also had antiviral and anti-proliferative activities comparable to the WHO International Standard, NIBSC 95/650, and the IFNA2 standard available from PBL Assay Science. Combining the expression and purification protocols developed here to produce IFNA2 on a laboratory scale with the commercial fermenter technology commonly used in pharmaceutical industry may further enhance IFNA2 yields, which will promote the development of interferon-based protein drugs to treat various disorders. PMID:24794500

  16. Bacterial challenges in food

    PubMed Central

    Collee, J. G.

    1974-01-01

    Qualitative and quantitative aspects of bacterial challenges that might be encountered in food are discussed with reference to recognized and relatively unrecognized hazards. Mechanisms of pathogenicity are reviewed and the populations at risk are noted. The bacterial content of food as it is served at table merits more study. The challenge of prevention by education is discussed. Indirect bacterial challenges in our food are considered. The real challenge of diagnosis depends upon an awareness of a complex range of conditions; the importance of effective communication with efficient laboratory and epidemiological services is stressed. There is an increasing need for care in the preparation and distribution of food. PMID:4467860

  17. Bacterial Wound Culture

    MedlinePlus

    ... Home Visit Global Sites Search Help? Bacterial Wound Culture Share this page: Was this page helpful? Also known as: Aerobic Wound Culture; Anaerobic Wound Culture Formal name: Culture, wound Related ...

  18. Bacterial Nail Infection (Paronychia)

    MedlinePlus

    ... in people who work in the health care industry. Chronic paronychia is most common in adult women and those who work in places where their hands are kept moist, such as food handlers. Signs and Symptoms Bacterial nail infection most ...

  19. Small bowel bacterial overgrowth

    MedlinePlus

    Overgrowth - intestinal bacteria; Bacterial overgrowth - intestine ... Unlike the large intestine, the small intestine does not have a high number of bacteria. When there are too many bacteria in the ...

  20. Bacterial surface adaptation

    NASA Astrophysics Data System (ADS)

    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.

  1. Body Basics

    MedlinePlus

    ... more about how the body works, what basic human anatomy is, and what happens when parts of the body don't function properly. Blood Bones, Muscles, and Joints Brain and Nervous System Digestive System Endocrine System Eyes Female Reproductive System ...

  2. Biochemical characterization and bacterial expression of an odorant-binding protein from Locusta migratoria.

    PubMed

    Ban, L; Scaloni, A; D'Ambrosio, C; Zhang, L; Yahn, Y; Pelosi, P

    2003-02-01

    Analysis of soluble proteins from different body parts of Locusta migratoria revealed a fast-migrating component in native electrophoresis, unique to antennae of both sexes. N-terminal sequence analysis and cloning identified this protein as a member of the insect odorant-binding proteins, carrying a well-conserved six-cysteine motif. Mass spectrometry analysis confirmed the occurrence of two distinct polypeptide species determined by nucleotide sequencing and demonstrated that the cysteine residues are paired in an interlocked fashion. The protein was expressed in a bacterial system with yields of about 10 mg/l of culture, mostly present as inclusion bodies. However, this recombinant product was solubilized after disulfide reduction. Air oxidation yielded a species with all disulfides spontaneously formed as in the native counterpart. Both native and recombinant proteins migrated as a dimer in gel filtration chromatography. Ligand binding was measured, using N-phenyl-1-naphthylamine as the fluorescent probe; the affinity of other ligands was measured in competitive binding assays. The protein exhibited great resistance to thermal denaturation even following prolonged treatment at 100 degrees C. A structural model for this dimeric species was generated on the basis of its sequence homology with Bombyx mori pheromone-binding protein, whose three-dimensional structure has been resolved as an unbound species and in complex with its physiological ligand. This is the first report of an odorant-binding protein identified and characterized from Orthoptera. PMID:12678502

  3. Deciphering the bacterial glycocode: recent advances in bacterial glycoproteomics

    PubMed Central

    Longwell, Scott A.; Dube, Danielle H.

    2012-01-01

    Bacterial glycoproteins represent an attractive target for new antibacterial treatments, as they are frequently linked to pathogenesis and contain distinctive glycans that are absent in humans. Despite their potential therapeutic importance, many bacterial glycoproteins remain uncharacterized. This review focuses on recent advances in deciphering the bacterial glycocode, including metabolic glycan labeling to discover and characterize bacterial glycoproteins, lectin-based microarrays to monitor bacterial glycoprotein dynamics, crosslinking sugars to assess the roles of bacterial glycoproteins, and harnessing bacterial glycosylation systems for the efficient production of industrially important glycoproteins. PMID:23276734

  4. Bistability and Bacterial Infections

    PubMed Central

    Malka, Roy; Shochat, Eliezer; Rom-Kedar, Vered

    2010-01-01

    Bacterial infections occur when the natural host defenses are overwhelmed by invading bacteria. The main component of the host defense is impaired when neutrophil count or function is too low, putting the host at great risk of developing an acute infection. In people with intact immune systems, neutrophil count increases during bacterial infection. However, there are two important clinical cases in which they remain constant: a) in patients with neutropenic-associated conditions, such as those undergoing chemotherapy at the nadir (the minimum clinically observable neutrophil level); b) in ex vivo examination of the patient's neutrophil bactericidal activity. Here we study bacterial population dynamics under fixed neutrophil levels by mathematical modelling. We show that under reasonable biological assumptions, there are only two possible scenarios: 1) Bacterial behavior is monostable: it always converges to a stable equilibrium of bacterial concentration which only depends, in a gradual manner, on the neutrophil level (and not on the initial bacterial level). We call such a behavior type I dynamics. 2) The bacterial dynamics is bistable for some range of neutrophil levels. We call such a behavior type II dynamics. In the bistable case (type II), one equilibrium corresponds to a healthy state whereas the other corresponds to a fulminant bacterial infection. We demonstrate that published data of in vitro Staphylococcus epidermidis bactericidal experiments are inconsistent with both the type I dynamics and the commonly used linear model and are consistent with type II dynamics. We argue that type II dynamics is a plausible mechanism for the development of a fulminant infection. PMID:20463954

  5. Women in Astronomy: Inclusion in Introductory Textbooks.

    ERIC Educational Resources Information Center

    Larsen, Kristine M.

    1995-01-01

    Discusses the reasons for identifying the contributions of women astronomers in introductory textbooks. The evolution of inclusion over the past 4 decades is explored by examining the rates of inclusion of 10 women astronomers in textbooks. Contains 100 references. (DDR)

  6. Forging Inclusive Solutions: Experiential Earth Charter Education

    ERIC Educational Resources Information Center

    Hill, Linda D.

    2010-01-01

    Forging Inclusive Solutions describes the aims, methodology and outcomes of Inclusive Leadership Adventures, an experiential education curriculum for exploring the Earth Charter. Experiential education builds meaningful relationships, skills, awareness and an inclusive community based on the Earth Charter principles. When we meet people where they…

  7. Deconstructing Normalisation: Clearing the Way for Inclusion.

    ERIC Educational Resources Information Center

    Culham, Andrew; Nind, Melanie

    2003-01-01

    This paper considers two major movements affecting the lives of people with intellectual disabilities: normalization and inclusion. It reviews the aims, processes, and outcomes of the normalization and social role valorization movement and explores its compatibility with inclusion. Lessons from normalization are applied to the inclusion movement.…

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

    ERIC Educational Resources Information Center

    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)

  9. Ultrastructural cytochemical analysis of intranuclear arsenic inclusions

    SciTech Connect

    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.

  10. Toward an Interdisciplinary Framework for Educational Inclusivity

    ERIC Educational Resources Information Center

    DeLuca, Christopher

    2013-01-01

    This purpose of this paper is to provide a theoretical foundation for inclusion in Canadian schools for this Special Issue on Inclusive Education. In response to the need for an interdisciplinary framework, this paper uses an interpretive literature review methodology to construct a framework for educational inclusivity based on four disciplinary…

  11. Louisiana High School Principals' Attitudes toward Inclusion

    ERIC Educational Resources Information Center

    Lorio, Katherine Matranga

    2011-01-01

    Principals' attitudes toward inclusion have been shown to have a positive impact on successful inclusive practices within schools. Providing guidance, support, and training to assist the faculty and staff with inclusion had been acknowledged to make a difference for schools administrators who were making changes to include more students with…

  12. Feature Issue on Inclusion and School Restructuring.

    ERIC Educational Resources Information Center

    Vandercook, Terri; York-Barr, Jennifer

    1996-01-01

    This feature issue on inclusion and school restructuring for students with and without disabilities contains the following articles: (1) "The 7 Habits for Educators" (Terri Vandercook); (2) "The Evolution of Inclusive Education" (Jennifer York-Barr and Terri Vandercook); (3) "Creating Inclusive Schools: What Does the Research Say?" (Margaret J.…

  13. Inclusive Education: Examining Equity on Five Continents

    ERIC Educational Resources Information Center

    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…

  14. Measuring Inclusive Education Outcomes in Alberta, Canada

    ERIC Educational Resources Information Center

    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…

  15. Ingredients for Inclusion: Lessons from the Literature

    ERIC Educational Resources Information Center

    McMaster, Christopher

    2012-01-01

    This paper explores features of successful inclusion through examples found in the literature. Schools have been given the imperative to become more inclusive through various government pronouncements and initiatives, but guidance in achieving that goal has been arguably wanting. School communities that have demonstrated more inclusive practice…

  16. Teachers' Experience with Inclusive Education in Singapore

    ERIC Educational Resources Information Center

    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.…

  17. Inclusive Attitudes: A Pre-Service Analysis

    ERIC Educational Resources Information Center

    Ryan, Thomas G.

    2009-01-01

    This enquiry has shown that pre-service teachers in Ontario, Canada (n = 141), believe they have the prerequisite training and knowledge to be inclusive educators. They believe inclusion is effective and produces required results for both identified and non-identified students. Inclusion was viewed as a difficult proposition dependent upon several…

  18. Elements of Inclusion: Findings from the Field

    ERIC Educational Resources Information Center

    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…

  19. Inclusion: By Choice or by Chance?

    ERIC Educational Resources Information Center

    Armstrong, Derrick; Armstrong, Ann Cheryl; Spandagou, Ilektra

    2011-01-01

    This paper explores the development internationally of the inclusive education perspective. Inclusive education as a late modernity reform project is exemplified in the call for "Education for All". Despite the simplicity of its message, inclusion is highly contestable. We argue in this paper that the key questions raised by the concept of…

  20. Initial insights into bacterial succession during human decomposition.

    PubMed

    Hyde, Embriette R; Haarmann, Daniel P; Petrosino, Joseph F; Lynne, Aaron M; Bucheli, Sibyl R

    2015-05-01

    Decomposition is a dynamic ecological process dependent upon many factors such as environment, climate, and bacterial, insect, and vertebrate activity in addition to intrinsic properties inherent to individual cadavers. Although largely attributed to microbial metabolism, very little is known about the bacterial basis of human decomposition. To assess the change in bacterial community structure through time, bacterial samples were collected from several sites across two cadavers placed outdoors to decompose and analyzed through 454 pyrosequencing and analysis of variable regions 3-5 of the bacterial 16S ribosomal RNA (16S rRNA) gene. Each cadaver was characterized by a change in bacterial community structure for all sites sampled as time, and decomposition, progressed. Bacteria community structure is variable at placement and before purge for all body sites. At bloat and purge and until tissues began to dehydrate or were removed, bacteria associated with flies, such as Ignatzschineria and Wohlfahrtimonas, were common. After dehydration and skeletonization, bacteria associated with soil, such as Acinetobacter, were common at most body sites sampled. However, more cadavers sampled through multiple seasons are necessary to assess major trends in bacterial succession. PMID:25431049

  1. Body lice

    MedlinePlus

    ... off the body. Your provider may prescribe a skin cream or a wash that contains permethrin, malathione, or benzyl alcohol. If your case is severe, the provider may prescribe medicine that you take by mouth.

  2. Body Image

    MedlinePlus

    ... spider veins Body dysmorphic disorder (BDD) Eating disorders Anorexia nervosa Binge eating disorder Bulimia nervosa Over-exercising ... conditions? Visit our Mental health section. Fact sheets Anorexia nervosa Binge eating disorder Bulimia nervosa Cosmetics and ...

  3. Body Signals.

    ERIC Educational Resources Information Center

    Gurley-Dilger, 'Laine

    1986-01-01

    Describes an activity in which students observe pairs of students engaged in conversation. Observations of "body language" are made, shifts in stance, and duration of stance. Discusses the application of this type of study to other species. (TW)

  4. Body lice

    MedlinePlus

    ... also get lice from infected clothing, towels, or bedding. Body lice are bigger than other types of ... last if you: Bathe regularly Wash clothes and bedding at least once a week Placing clothes in ...

  5. Bog bodies.

    PubMed

    Lynnerup, Niels

    2015-06-01

    In northern Europe during the Iron Age, many corpses were deposited in bogs. The cold, wet and anaerobic environment leads in many cases to the preservation of soft tissues, so that the bodies, when found and excavated several thousand years later, are remarkably intact. Since the 19th century the bog bodies have been studied using medical and natural scientific methods, and recently many bog bodies have been re-examined using especially modern, medical imaging techniques. Because of the preservation of soft tissue, especially the skin, it has been possible to determine lesions and trauma. Conversely, the preservation of bones is less good, as the mineral component has been leached out by the acidic bog. Together with water-logging of collagenous tissue, this means that if the bog body is simply left to dry out when found, as was the case pre-19th century, the bones may literally warp and shrink, leading to potential pitfalls in paleopathological diagnostics. Bog bodies have in several instances been crucial in determining the last meal, as gut contents may be preserved, and thus augment our knowledge on pre-historic diet by adding to, for example, stable isotope analyses. This article presents an overview of our knowledge about the taphomic processes as well as the methods used in bog body research. PMID:25998635

  6. On the problem of a thin rigid inclusion embedded in a Maxwell material

    NASA Astrophysics Data System (ADS)

    Popova, T.; Rogerson, G. A.

    2016-08-01

    We consider a plane viscoelastic body, composed of Maxwell material, with a crack and a thin rigid inclusion. The statement of the problem includes boundary conditions in the form of inequalities, together with an integral condition describing the equilibrium conditions of the inclusion. An equivalent variational statement is provided and used to prove the uniqueness of the problem's solution. The analysis is carried out in respect of perfect and non-perfect bonding of the rigid inclusion. Additional smoothness properties of the solutions, namely the existence of the time derivative, are also established.

  7. Bacterial start site prediction.

    PubMed

    Hannenhalli, S S; Hayes, W S; Hatzigeorgiou, A G; Fickett, J W

    1999-09-01

    With the growing number of completely sequenced bacterial genes, accurate gene prediction in bacterial genomes remains an important problem. Although the existing tools predict genes in bacterial genomes with high overall accuracy, their ability to pinpoint the translation start site remains unsatisfactory. In this paper, we present a novel approach to bacterial start site prediction that takes into account multiple features of a potential start site, viz., ribosome binding site (RBS) binding energy, distance of the RBS from the start codon, distance from the beginning of the maximal ORF to the start codon, the start codon itself and the coding/non-coding potential around the start site. Mixed integer programing was used to optimize the discriminatory system. The accuracy of this approach is up to 90%, compared to 70%, using the most common tools in fully automated mode (that is, without expert human post-processing of results). The approach is evaluated using Bacillus subtilis, Escherichia coli and Pyrococcus furiosus. These three genomes cover a broad spectrum of bacterial genomes, since B.subtilis is a Gram-positive bacterium, E.coli is a Gram-negative bacterium and P. furiosus is an archaebacterium. A significant problem is generating a set of 'true' start sites for algorithm training, in the absence of experimental work. We found that sequence conservation between P. furiosus and the related Pyrococcus horikoshii clearly delimited the gene start in many cases, providing a sufficient training set. PMID:10446249

  8. Questioning Secondary Inclusive Education: Are Inclusive Classrooms Always Best for Students?

    ERIC Educational Resources Information Center

    Tkachyk, Ruth Elizabeth

    2013-01-01

    Educating students with special needs in inclusive settings has become a priority for westernized governments as they strive to create more inclusive societies. While recognizing the societal benefits of inclusion, teachers and parents question whether or not implementation of full inclusion will come at the expense of learners' individual…

  9. Measuring Quality in Inclusive Preschool Classrooms: Development and Validation of the Inclusive Classroom Profile (ICP)

    ERIC Educational Resources Information Center

    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. Melt inclusions in Luna 24 soil fragments

    NASA Technical Reports Server (NTRS)

    Roedder, W.; Weiblen, P. W.

    1978-01-01

    Optical examinations of 28 slides of Luna 24 soil fragments revealed melt inclusions in grains of olivine, plagioclase, spinel, and ilmenite as well as interstitial inclusions. In contrast with Apollo samples, the Luna 24 samples contain sulfide melt inclusions, which indicates that saturation with respect to an iron sulfide melt took place throughout much of the crystallization history, even while olivine was crystallizing. The Luna 24 silicate-melt inclusions have recorded a more extensive differentiation toward higher iron magmas than have the Apollo inclusions, but they have also recorded some inexplicably low aluminum values.

  11. Lipoproteins of bacterial pathogens.

    PubMed

    Kovacs-Simon, A; Titball, R W; Michell, S L

    2011-02-01

    Bacterial lipoproteins are a set of membrane proteins with many different functions. Due to this broad-ranging functionality, these proteins have a considerable significance in many phenomena, from cellular physiology through cell division and virulence. Here we give a general overview of lipoprotein biogenesis and highlight examples of the roles of lipoproteins in bacterial disease caused by a selection of medically relevant Gram-negative and Gram-positive pathogens: Mycobacterium tuberculosis, Streptococcus pneumoniae, Borrelia burgdorferi, and Neisseria meningitidis. Lipoproteins have been shown to play key roles in adhesion to host cells, modulation of inflammatory processes, and translocation of virulence factors into host cells. As such, a number of lipoproteins have been shown to be potential vaccines. This review provides a summary of some of the reported roles of lipoproteins and of how this knowledge has been exploited in some cases for the generation of novel countermeasures to bacterial diseases. PMID:20974828

  12. Distribution of fluid inclusions in igneous quartz of the Capitan pluton, New Mexico, USA

    NASA Astrophysics Data System (ADS)

    Ratajeski, Kent; Campbell, Andrew R.

    1994-02-01

    Fluid inclusions in granites have been used extensively in studies of magmatic-hydrothermal processes, but few studies have documented the distribution of secondary fluid inclusions within the nonmineralized main body of an intrusion. Previous work on fluid inclusions in Th-U-REE quartz/ fluorite vein deposits in the Capitan pluton suggests that these veins resulted from high-temperature, high-salinity, magmatic fluids in fractured and brecciated zones in the cooled outer carapace of the pluton. Petrographic examination of phenocryst quartz in forty-four thin sections of granite from the Capitan pluton reveals that these same fluids are found in secondary fluid inclusions. Fluid inclusions with daughter minerals commonly occur in the outer granophyre and aplite zones, but are usually absent in the porphyritic core. Microthermometric data and calculated salinities from fifty-eight fluid inclusions in phenocryst quartz and thirty-one fluid inclusions in miarolitic quartz (both samples taken from the granophyre zone) also correlate fairly well with the data from the Th-U-REE quartz/fluorite vein deposits, and similar types of fluid inclusions are found in all three occurrences of quartz. A major difference, however, is that vapor-rich inclusions, abundant as secondary inclusions in igneous phenocryst quartz, are extremely scarce in the vein quartz. Liquid-vapor homogenization temperatures of high-salinity inclusions in phenocryst quartz range from 204-509°C with an average of 384°C. Halite dissolution temperatures of high salinity inclusions range from 442-583°C with an average of 525°C. Total homogenization temperatures are not as well documented due to decrepitation of many of the high salinity inclusions before total homogenization. Samples of miarolitic quartz appear to contain more low-salinity fluid inclusions than the phenocryst quartz, but high-salinity inclusions are present as well. These observations suggest that the magmatic-hydrothermal fluids

  13. Antiferromagnetic inclusions in lunar glass

    USGS Publications Warehouse

    Thorpe, A.N.; Senftle, F.E.; Briggs, Charles; Alexander, Corrine

    1974-01-01

    The magnetic susceptibility of 11 glass spherules from the Apollo 15, 16, and 17 fines and two specimens of a relatively large glass spherical shell were studied as a function of temperature from room temperature to liquid helium temperatures. All but one specimen showed the presence of antiferromagnetic inclusions. Closely spaced temperature measurements of the magnetic susceptibility below 77 K on five of the specimens showed antiferromagnetic temperature transitions (Ne??el transitions). With the exception of ilmenite in one specimen, these transitions did not correspond to any transitions in known antiferromagnetic compounds. ?? 1974.

  14. Transverse SSA in inclusive DIS

    NASA Astrophysics Data System (ADS)

    Pitonyak, Daniel

    2013-10-01

    We analyze the transverse single spin asymmetry (SSA) in inclusive deep inelastic scattering (DIS), which requires a two-photon exchange to generate a non-zero effect. We present numerical results for the SSA that allow us to comment on the so-called "sign mismatch" issue invloving the Efremov-Teryaev-Qiu-Sterman (ETQS) function TF(x,x). In particular, we discuss how our results indicate a collinear twist-3 Sivers-type effect may not be the main cause of the SSAs seen in proton-proton (pp) collisions.

  15. Inclusive fitness theory and eusociality.

    PubMed

    Abbot, Patrick; Abe, Jun; Alcock, John; Alizon, Samuel; Alpedrinha, Joao A C; Andersson, Malte; Andre, Jean-Baptiste; van Baalen, Minus; Balloux, Francois; Balshine, Sigal; Barton, Nick; Beukeboom, Leo W; Biernaskie, Jay M; Bilde, Trine; Borgia, Gerald; Breed, Michael; Brown, Sam; Bshary, Redouan; Buckling, Angus; Burley, Nancy T; Burton-Chellew, Max N; Cant, Michael A; Chapuisat, Michel; Charnov, Eric L; Clutton-Brock, Tim; Cockburn, Andrew; Cole, Blaine J; Colegrave, Nick; Cosmides, Leda; Couzin, Iain D; Coyne, Jerry A; Creel, Scott; Crespi, Bernard; Curry, Robert L; Dall, Sasha R X; Day, Troy; Dickinson, Janis L; Dugatkin, Lee Alan; El Mouden, Claire; Emlen, Stephen T; Evans, Jay; Ferriere, Regis; Field, Jeremy; Foitzik, Susanne; Foster, Kevin; Foster, William A; Fox, Charles W; Gadau, Juergen; Gandon, Sylvain; Gardner, Andy; Gardner, Michael G; Getty, Thomas; Goodisman, Michael A D; Grafen, Alan; Grosberg, Rick; Grozinger, Christina M; Gouyon, Pierre-Henri; Gwynne, Darryl; Harvey, Paul H; Hatchwell, Ben J; Heinze, Jürgen; Helantera, Heikki; Helms, Ken R; Hill, Kim; Jiricny, Natalie; Johnstone, Rufus A; Kacelnik, Alex; Kiers, E Toby; Kokko, Hanna; Komdeur, Jan; Korb, Judith; Kronauer, Daniel; Kümmerli, Rolf; Lehmann, Laurent; Linksvayer, Timothy A; Lion, Sébastien; Lyon, Bruce; Marshall, James A R; McElreath, Richard; Michalakis, Yannis; Michod, Richard E; Mock, Douglas; Monnin, Thibaud; Montgomerie, Robert; Moore, Allen J; Mueller, Ulrich G; Noë, Ronald; Okasha, Samir; Pamilo, Pekka; Parker, Geoff A; Pedersen, Jes S; Pen, Ido; Pfennig, David; Queller, David C; Rankin, Daniel J; Reece, Sarah E; Reeve, Hudson K; Reuter, Max; Roberts, Gilbert; Robson, Simon K A; Roze, Denis; Rousset, Francois; Rueppell, Olav; Sachs, Joel L; Santorelli, Lorenzo; Schmid-Hempel, Paul; Schwarz, Michael P; Scott-Phillips, Tom; Shellmann-Sherman, Janet; Sherman, Paul W; Shuker, David M; Smith, Jeff; Spagna, Joseph C; Strassmann, Beverly; Suarez, Andrew V; Sundström, Liselotte; Taborsky, Michael; Taylor, Peter; Thompson, Graham; Tooby, John; Tsutsui, Neil D; Tsuji, Kazuki; Turillazzi, Stefano; Ubeda, Francisco; Vargo, Edward L; Voelkl, Bernard; Wenseleers, Tom; West, Stuart A; West-Eberhard, Mary Jane; Westneat, David F; Wiernasz, Diane C; Wild, Geoff; Wrangham, Richard; Young, Andrew J; Zeh, David W; Zeh, Jeanne A; Zink, Andrew

    2011-03-24

    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. PMID:21430721

  16. Bacterial transfer RNAs

    PubMed Central

    Shepherd, Jennifer; Ibba, Michael

    2015-01-01

    Transfer RNA is an essential adapter molecule that is found across all three domains of life. The primary role of transfer RNA resides in its critical involvement in the accurate translation of messenger RNA codons during protein synthesis and, therefore, ultimately in the determination of cellular gene expression. This review aims to bring together the results of intensive investigations into the synthesis, maturation, modification, aminoacylation, editing and recycling of bacterial transfer RNAs. Codon recognition at the ribosome as well as the ever-increasing number of alternative roles for transfer RNA outside of translation will be discussed in the specific context of bacterial cells. PMID:25796611

  17. A Ruptured Digital Epidermal Inclusion Cyst: A Sinister Presentation

    PubMed Central

    Fletcher, Phillip; Ragg, Amanda; Vane, Andrew

    2016-01-01

    Epidermal inclusion cysts are benign cutaneous lesions caused by dermal or subdermal implantation and proliferation of epidermal squamous epithelium as a result of trauma or surgery. They are typically located on the scalp, face, trunk, neck, or back; however they can be found anywhere on the body. Lesions are asymptomatic unless complicated by rupture, malignant transformation to squamous cell carcinoma, or infection at which point they can clinically appear as more sinister pathologies. We present the case of a 45-year-old laborer with a ruptured epidermal inclusion cyst, manifesting clinically and radiographically as a malignancy. Following MRI, definitive surgical management may appear to be a logical progression in management of the patient. This case however is a good example of why meticulously following surgical protocol when evaluating an unknown soft tissue mass is imperative. By following protocol, an alternate diagnosis was made and the patient has since gone on to a make a full recovery without life transforming surgery. PMID:27418992

  18. A Ruptured Digital Epidermal Inclusion Cyst: A Sinister Presentation.

    PubMed

    Bohler, Iain; Fletcher, Phillip; Ragg, Amanda; Vane, Andrew

    2016-01-01

    Epidermal inclusion cysts are benign cutaneous lesions caused by dermal or subdermal implantation and proliferation of epidermal squamous epithelium as a result of trauma or surgery. They are typically located on the scalp, face, trunk, neck, or back; however they can be found anywhere on the body. Lesions are asymptomatic unless complicated by rupture, malignant transformation to squamous cell carcinoma, or infection at which point they can clinically appear as more sinister pathologies. We present the case of a 45-year-old laborer with a ruptured epidermal inclusion cyst, manifesting clinically and radiographically as a malignancy. Following MRI, definitive surgical management may appear to be a logical progression in management of the patient. This case however is a good example of why meticulously following surgical protocol when evaluating an unknown soft tissue mass is imperative. By following protocol, an alternate diagnosis was made and the patient has since gone on to a make a full recovery without life transforming surgery. PMID:27418992

  19. A corundum-rich inclusion in the Murchison carbonaceous chondrite

    NASA Astrophysics Data System (ADS)

    Bar-Matthews, M.; Hutcheon, I. D.; MacPherson, G. J.; Grossman, L.

    1982-01-01

    Although thermodynamic calculations predict corundum to be the first condensate from a cooling gas of solar composition, a corundum-hibonite inclusion, BB-5, has for the first time been found in the Murchison carbonaceous chondrite. Ion microprobe measurements of Mg isotopic compositions yield the result, unexpected in such an early condensate, that Mg-26 excesses are small despite large Al-27/Mg-24 ratios. The extreme temperature required to melt this inclusion makes a liquid origin unlikely, except by the hypervelocity impact of refractory bodies. Alternatively, B-5 is a direct gas-solid condensate, and its uniform Mg-26 enrichment must be a characteristic of the reservoir from which it condensed. Nebular heterogeneity in magnesium isotopic composition is the preferred explanation for the formation of such a reservoir.

  20. Body Imaging

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The high-tech art of digital signal processing (DSP) was pioneered at NASA's Jet Propulsion Laboratory (JPL) in the mid-1960s for use in the Apollo Lunar Landing Program. Designed to computer enhance pictures of the Moon, this technology became the basis for the Landsat Earth resources satellites and subsequently has been incorporated into a broad range of Earthbound medical and diagnostic tools. DSP is employed in advanced body imaging techniques including Computer-Aided Tomography, also known as CT and CATScan, and Magnetic Resonance Imaging (MRI). CT images are collected by irradiating a thin slice of the body with a fan-shaped x-ray beam from a number of directions around the body's perimeter. A tomographic (slice-like) picture is reconstructed from these multiple views by a computer. MRI employs a magnetic field and radio waves, rather than x-rays, to create images. In this photograph, a patient undergoes an open MRI.

  1. Body Imaging

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The high-tech art of digital signal processing (DSP) was pioneered at NASA's Jet Propulsion Laboratory (JPL) in the mid-1960s for use in the Apollo Lunar Landing Program. Designed to computer enhance pictures of the Moon, this technology became the basis for the Landsat Earth resources satellites and subsequently has been incorporated into a broad range of Earthbound medical and diagnostic tools. DSP is employed in advanced body imaging techniques including Computer-Aided Tomography, also known as CT and CATScan, and Magnetic Resonance Imaging (MRI). CT images are collected by irradiating a thin slice of the body with a fan-shaped x-ray beam from a number of directions around the body's perimeter. A tomographic (slice-like) picture is reconstructed from these multiple views by a computer. MRI employs a magnetic field and radio waves, rather than x-rays, to create images.

  2. Body Imaging

    NASA Technical Reports Server (NTRS)

    1990-01-01

    Magnetic Resonance Imaging (MRI) and Computer-aided Tomography (CT) images are often complementary. In most cases, MRI is good for viewing soft tissue but not bone, while CT images are good for bone but not always good for soft tissue discrimination. Physicians and engineers in the Department of Radiology at the University of Michigan Hospitals are developing a technique for combining the best features of MRI and CT scans to increase the accuracy of discriminating one type of body tissue from another. One of their research tools is a computer program called HICAP. The program can be used to distinguish between healthy and diseased tissue in body images.

  3. Semi-inclusive DIS: Factorization

    SciTech Connect

    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.

  4. Topological derivatives for fundamental frequencies of elastic bodies

    NASA Astrophysics Data System (ADS)

    Kobelev, Vladimir

    2016-01-01

    In this article a new method for topological optimization of fundamental frequencies of elastic bodies, which could be considered as an improvement on the bubble method, is introduced. The method is based on generalized topological derivatives. For a body with different types of inclusion the vector genus is introduced. The dimension of the genus is the number of different elastic properties of the inclusions being introduced. The disturbances of stress and strain fields in an elastic matrix due to a newly inserted elastic inhomogeneity are given explicitly in terms of the stresses and strains in the initial body. The iterative positioning of inclusions is carried out by determination of the preferable position of the new inhomogeneity at the extreme points of the characteristic function. The characteristic function was derived using Eshelby's method. The expressions for optimal ratios of the semi-axes of the ellipse and angular orientation of newly inserted infinitesimally small inclusions of elliptical form are derived in closed analytical form.

  5. Bacterial porins stimulate bone resorption.

    PubMed Central

    Meghji, S; Henderson, B; Nair, S P; Tufano, M A

    1997-01-01

    Porins are abundant outer membrane proteins of gram-negative bacteria involved in transport of low-molecular-mass molecules. During the past decade, porins from a number of bacteria have also been shown to have proinflammatory activities including inducing the synthesis of proinflammatory mediators (cytokines, platelet-activating factor, and nitric oxide) in cultured cells and inducing inflammation in vivo. With this range of actions, it was possible that porins could also interact with bone cells to cause aberrant bone remodeling and that this could contribute to the bone destruction seen in gram-negative bone infections. By using purified preparations of Salmonella typhimurium and Pseudomonas aeruginosa porins, in the presence of polymyxin B, it was possible to induce concentration-dependent loss of calcium from cultured murine calvaria at porin concentrations in the range of 1 to 10 nM. The mechanism of action of the porins was determined by the inclusion of inhibitors of cyclooxygenase or inflammatory cytokines in the culture media. The bone-resorbing activity of both porins was not inhibited by the cyclooxygenase inhibitor indomethacin or by neutralizing the activity of tumor necrosis factor. Indeed, relatively high concentrations of these agents produced an unexpected increase in the bone resorption induced by the porins. In contrast, porin-induced bone resorption could be inhibited by relatively high concentrations of the natural inhibitor of interleukin-1 (IL-1 receptor antagonist). It appears that these porins stimulate bone resorption by a mechanism distinct from that of lipopolysaccharide, and the possibility therefore exists that porins play a role in bone destruction in gram-negative bacterial infections of bone. PMID:9119467

  6. Modeling intraocular bacterial infections.

    PubMed

    Astley, Roger A; Coburn, Phillip S; Parkunan, Salai Madhumathi; Callegan, Michelle C

    2016-09-01

    Bacterial endophthalmitis is an infection and inflammation of the posterior segment of the eye which can result in significant loss of visual acuity. Even with prompt antibiotic, anti-inflammatory and surgical intervention, vision and even the eye itself may be lost. For the past century, experimental animal models have been used to examine various aspects of the pathogenesis and pathophysiology of bacterial endophthalmitis, to further the development of anti-inflammatory treatment strategies, and to evaluate the pharmacokinetics and efficacies of antibiotics. Experimental models allow independent control of many parameters of infection and facilitate systematic examination of infection outcomes. While no single animal model perfectly reproduces the human pathology of bacterial endophthalmitis, investigators have successfully used these models to understand the infectious process and the host response, and have provided new information regarding therapeutic options for the treatment of bacterial endophthalmitis. This review highlights experimental animal models of endophthalmitis and correlates this information with the clinical setting. The goal is to identify knowledge gaps that may be addressed in future experimental and clinical studies focused on improvements in the therapeutic preservation of vision during and after this disease. PMID:27154427

  7. Phagocytosis of bacterial pathogens.

    PubMed

    Chung, Yoon-Suk Alexander; Kocks, Christine

    2012-01-01

    Phagocytosis is an evolutionarily ancient, receptor-driven process, by which phagocytic cells recognize invading microbes and destroy them after internalization. The phagocytosis receptor Eater is expressed exclusively on Drosophila phagocytes and is required for the survival of bacterial infections. In a recent study, we explored how Eater can defend fruit flies against different kinds of bacteria. We discovered that Eater bound to certain types of bacteria directly, while for others bacterial binding was dependent on prior disruption of the bacterial envelope. Similar to phagocytes, antimicrobial peptides and lysozymes are ancient components of animal immune systems. Our results suggest that cationic antimicrobial peptides, as well as lysozymes, can facilitate Eater binding to live Gram-negative bacteria. Both types of molecules promote surface-exposure of bacterial ligands that otherwise would remain buried and hidden under an outer membrane. We propose that unmasking ligands for phagocytic receptors may be a conserved mechanism operating in many animals, including humans. Thus, studying a Drosophila phagocytosis receptor may advance our understanding of innate immunity in general. PMID:22223092

  8. Bacterial extracellular lignin peroxidase

    DOEpatents

    Crawford, Donald L.; Ramachandra, Muralidhara

    1993-01-01

    A newly discovered lignin peroxidase enzyme is provided. The enzyme is obtained from a bacterial source and is capable of degrading the lignin portion of lignocellulose in the presence of hydrogen peroxide. The enzyme is extracellular, oxidative, inducible by lignin, larch wood xylan, or related substrates and capable of attacking certain lignin substructure chemical bonds that are not degradable by fungal lignin peroxidases.

  9. BACTERIAL WATERBORNE PATHOGENS

    EPA Science Inventory

    Bacterial pathogens are examples of classical etiological agents of waterborne disease. While these agents no longer serve as major threats to U.S. water supplies, they are still important pathogens in areas with substandard sanitation and poor water treatment facilities. In th...

  10. Bacterial microflora of nectarines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Microflora of fruit surfaces has been the best source of antagonists against fungi causing postharvest decays of fruit. However, there is little information on microflora colonizing surfaces of fruits other than grapes, apples, and citrus fruit. We characterized bacterial microflora on nectarine f...

  11. Bacterial leaf spot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial leaf spot has been reported in Australia (Queensland), Egypt, El Salvador, India, Japan, Nicaragua, Sudan, and the United States (Florida, Iowa, Kansas, Maryland, and Wisconsin). It occasionally causes locally severe defoliation and post-emergence damping-off and stunting. The disease is...

  12. The Bacterial Growth Curve.

    ERIC Educational Resources Information Center

    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)

  13. Pulmonary neuroendocrine tumors with nuclear inclusion.

    PubMed

    Kobayashi, Saori; Tsuta, Koji; Sekine, Shigeki; Yoshida, Akihiko; Sasaki, Naoshi; Shibuki, Yasuo; Sakurai, Hiroyuki; Watanabe, Shun-Ichi; Asamura, Hisao; Tsuda, Hitoshi

    2013-09-01

    Nuclear inclusion or pseudoinclusion is a peculiar cytological feature, and its recognition in appropriate clinicopathological settings can aid in the diagnosis of several disease entities. To the best of our knowledge, only 1 case of pulmonary neuroendocrine tumor (NET) with nuclear pseudoinclusion has been reported. A review of 227 patients who had undergone surgical resection for pulmonary NETs revealed 2 tumors with different mechanisms of nuclear inclusion. To explore the cause of nuclear inclusion, NET with nuclear inclusion was characterized immunohistochemically and ultrastructurally. Nuclear inclusions were observed in 2 of the 227 (0.9%) patients with pulmonary NETs. The first patient was a 46-year-old woman with small cell carcinoma. Tumor cells with nuclear inclusions were distributed focally. Ultrastructural analysis showed that these inclusions were pseudoinclusions. The second patient was a 62-year-old man with large-cell neuroendocrine carcinoma. Nuclear inclusions were observed in the focal area of the tumor. Immunohistochemical analysis revealed that the intra-nuclear materials consisted of biotin and aberrant cytoplasmic and nuclear accumulation of β-catenin. Mutational analysis revealed a CTNNB1 gene mutation. Although very rare, diagnostic errors may be observed in cases of pulmonary NETs with nuclear inclusions. The mechanisms of nuclear inclusion differed, with one due to herniation of the cytoplasm into the nucleus (pseudoinclusion) and the other due to accumulation of biotin resulting from a CTNNB1 gene mutation. PMID:23896262

  14. Body Levers.

    ERIC Educational Resources Information Center

    Chiappetta, Eugene L.

    1987-01-01

    Explains how the muscle and bone arrangement of the human body can be used in teaching the principles of simple machines. Presents an activity that investigates the lever system of the forearm. Includes background information on levers and suggests questions for classroom discussion. (ML)

  15. Corticosteroids for Bacterial Keratitis

    PubMed Central

    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

  16. A case of recalcitrant bacterial conjunctivitis.

    PubMed

    Aboshiha, Jonathan

    2013-01-01

    It is important to be vigilant for retained foreign bodies as a cause of recalcitrant bacterial conjunctivitis, even in the absence of foreign body sensation. A relapsing-remitting history should prompt referral to an ophthalmology department. All patients presenting with a red eye should be asked specifically about contact lens wear, and causes of conjunctivitis other than those bacterial in nature--such as viral and chlamydial infections or allergy--should be borne in mind. Contact lens wear may cause ocular complications, ranging from mild dry eye symptoms to contact lens-associated microbial keratitis, which is an ophthalmic emergency. Contact lens-associated corneal infections caused by Pseudomonas aeruginosa, which can rapidly penetrate the cornea, or Acanthamoeba can be severe and sight threatening. All patients with a history of contact lens wear and red flag symptoms such as eye pain, redness, reduction or change in vision, corneal epithelial defect, discharge, foreign body sensation or failure to respond to antibiotics should be referred for an urgent ophthalmic review. Retained contact lenses are known to cause several ocular complications, such as giant papillary conjunctivitis and ulcerative keratitis which may threaten corneal penetration. Lid eversion may reveal a hidden sub-tarsal contact lens. Use of fluorescein may also allow visualisation of any corneal epithelial defect. A careful slit lamp examination by an ophthalmologist is required to exclude this critical finding definitively. PMID:24555257

  17. [Body contouring].

    PubMed

    Krüger, N; Lübberding, S; Sattler, G

    2015-10-01

    In our contemporary postmodern society, a modified perception of the human body is accompanied by an increasing demand for body shaping procedures. The treatment needs to be effective but it is just as important that they are safe and can be easily integrated into the daily working and routine schedule. While the options for minimally invasive volume addition are largely limited to injectable implants based on hyaluronic acid or autologous fat, a multitude of options are available for volume reduction. Before deciding on the method of choice, the following needs to be considered: which indications need to be treated, the extent of the reduction in volume and how much pain and possible undesired reactions the patient is prepared to accept. PMID:26349684

  18. Making the case for inclusive design.

    PubMed

    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. PMID:23538129

  19. Dementia with Lewy bodies in Meige Syndrome

    PubMed Central

    Ho, Ho Yin; Yu, Chi Shing

    2012-01-01

    Meige syndrome is a rare form of segmental dystonia characterized by blepharospasm and oromandibular dystonia. A few case reports of Meige syndrome have been associated with Lewy body pathologies, and the syndrome was also proposed for inclusion in the spectrum of Lewy body disease. We report a case of an elderly gentleman with a history of Meige syndrome for more than 10 years who developed dementia with Lewy bodies. Updated clinical and pathological evidence of linkages between these two conditions is also presented. PMID:22984651

  20. Bacterial interference with canonical NFκB signalling

    PubMed Central

    Askarian, Fatemeh; Sangvik, Maria; Sollid, Johanna E.

    2013-01-01

    The human body is constantly challenged by a variety of commensal and pathogenic micro-organisms that trigger the immune system. Central in the first line of defence is the pattern-recognition receptor (PRR)-induced stimulation of the NFκB pathway, leading to NFκB activation. The subsequent production of pro-inflammatory cytokines and/or antimicrobial peptides results in recruitment of professional phagocytes and bacterial clearance. To overcome this, bacteria have developed mechanisms for targeted interference in every single step in the PRR–NFκB pathway to dampen host inflammatory responses. This review aims to briefly overview the PRR–NFκB pathway in relation to the immune response and give examples of the diverse bacterial evasion mechanisms including changes in the bacterial surface, decoy production and injection of effector molecules. Targeted regulation of inflammatory responses is needed and bacterial molecules developed for immune evasion could provide future anti-inflammatory agents. PMID:23873783

  1. Detection of Bacterial Spores with Lanthanide-Macrocycle Binary Complexes

    PubMed Central

    Cable, Morgan L.; Kirby, James P.; Levine, Dana J.; Manary, Micah J.; Gray, Harry B.; Ponce, Adrian

    2009-01-01

    The detection of bacterial spores via dipicolinate-triggered lanthanide luminescence has been improved in terms of detection limit, stability, and susceptibility to interferents by use of lanthanide-macrocycle binary complexes. Specifically, we compared the effectiveness of Sm, Eu, Tb and Dy complexes with the macrocycle 1,4,7,10-tetraazacyclododecane-1,7-diacetate (DO2A) to the corresponding lanthanide aquo ions. The Ln(DO2A)+ binary complexes bind dipicolinic acid (DPA), a major constituent of bacterial spores, with greater affinity and demonstrate significant improvement in bacterial spore detection. Of the four luminescent lanthanides studied, the terbium complex exhibits the greatest dipicolinate binding affinity (100-fold greater than Tb3+ alone, and 10-fold greater than other Ln(DO2A)+ complexes) and highest quantum yield. Moreover, the inclusion of DO2A extends the pH range over which Tb-DPA coordination is stable, reduces the interference of calcium ions nearly 5-fold, and mitigates phosphate interference 1000-fold compared to free terbium alone. In addition, detection of Bacillus atrophaeus bacterial spores was improved by the use of Tb(DO2A)+, yielding a 3-fold increase in the signal-to-noise ratio over Tb3+. Out of the eight cases investigated, the Tb(DO2A)+ binary complex is best for the detection of bacterial spores. PMID:19537757

  2. Bacterial Genome Instability

    PubMed Central

    Darmon, Elise

    2014-01-01

    SUMMARY Bacterial genomes are remarkably stable from one generation to the next but are plastic on an evolutionary time scale, substantially shaped by horizontal gene transfer, genome rearrangement, and the activities of mobile DNA elements. This implies the existence of a delicate balance between the maintenance of genome stability and the tolerance of genome instability. In this review, we describe the specialized genetic elements and the endogenous processes that contribute to genome instability. We then discuss the consequences of genome instability at the physiological level, where cells have harnessed instability to mediate phase and antigenic variation, and at the evolutionary level, where horizontal gene transfer has played an important role. Indeed, this ability to share DNA sequences has played a major part in the evolution of life on Earth. The evolutionary plasticity of bacterial genomes, coupled with the vast numbers of bacteria on the planet, substantially limits our ability to control disease. PMID:24600039

  3. Bacterial multidrug efflux transporters.

    PubMed

    Delmar, Jared A; Su, Chih-Chia; Yu, Edward W

    2014-01-01

    Infections caused by bacteria are a leading cause of death worldwide. Although antibiotics remain a key clinical therapy, their effectiveness has been severely compromised by the development of drug resistance in bacterial pathogens. Multidrug efflux transporters--a common and powerful resistance mechanism--are capable of extruding a number of structurally unrelated antimicrobials from the bacterial cell, including antibiotics and toxic heavy metal ions, facilitating their survival in noxious environments. Transporters of the resistance-nodulation-cell division (RND) superfamily typically assemble as tripartite efflux complexes spanning the inner and outer membranes of the cell envelope. In Escherichia coli, the CusCFBA complex, which mediates resistance to copper(I) and silver(I) ions, is the only known RND transporter specific to heavy metals. Here, we describe the current knowledge of individual pump components of the Cus system, a paradigm for efflux machinery, and speculate on how RND pumps assemble to fight diverse antimicrobials. PMID:24702006

  4. Physics of Bacterial Morphogenesis

    PubMed Central

    Sun, Sean X.; Jiang, Hongyuan

    2011-01-01

    Summary: Bacterial cells utilize three-dimensional (3D) protein assemblies to perform important cellular functions such as growth, division, chemoreception, and motility. These assemblies are composed of mechanoproteins that can mechanically deform and exert force. Sometimes, small-nucleotide hydrolysis is coupled to mechanical deformations. In this review, we describe the general principle for an understanding of the coupling of mechanics with chemistry in mechanochemical systems. We apply this principle to understand bacterial cell shape and morphogenesis and how mechanical forces can influence peptidoglycan cell wall growth. We review a model that can potentially reconcile the growth dynamics of the cell wall with the role of cytoskeletal proteins such as MreB and crescentin. We also review the application of mechanochemical principles to understand the assembly and constriction of the FtsZ ring. A number of potential mechanisms are proposed, and important questions are discussed. PMID:22126993

  5. Formation of bacterial nanocells

    NASA Astrophysics Data System (ADS)

    Vainshtein, Mikhail; Kudryashova, Ekaterina; Suzina, Natalia; Ariskina, Elena; Voronkov, Vadim

    1998-07-01

    Existence of nanobacteria received increasing attention both in environmental microbiology/geomicro-biology and in medical microbiology. In order to study a production of nanoforms by typical bacterial cells. Effects of different physical factors were investigated. Treatment of bacterial cultures with microwave radiation, or culturing in field of electric current resulted in formation a few types of nanocells. The number and type of nanoforms were determined with type and dose of the treatment. The produced nanoforms were: i) globules, ii) clusters of the globules--probably produced by liaison, iii) nanocells coated with membrane. The viability of the globules is an object opened for doubts. The nanocells discovered multiplication and growth on solidified nutrient media. The authors suggest that formation of nanocells is a common response of bacteria to stress-actions produced by different agents.

  6. Bacterial Cell Wall Components

    NASA Astrophysics Data System (ADS)

    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.

  7. [Bacterial diseases of rape].

    PubMed

    Zakharova, O M; Mel'nychuk, M D; Dankevych, L A; Patyka, V P

    2012-01-01

    Bacterial destruction of the culture was described and its agents identified in the spring and winter rape crops. Typical symptoms are the following: browning of stem tissue and its mucilagization, chlorosis of leaves, yellowing and beginning of soft rot in the place of leaf stalks affixion to stems, loss of pigmentation (violet). Pathogenic properties of the collection strains and morphological, cultural, physiological, and biochemical properties of the agents of rape's bacterial diseases isolated by the authors have been investigated. It was found that all the isolates selected by the authors are highly or moderately aggressive towards different varieties of rape. According to the complex of phenotypic properties 44% of the total number of isolates selected by the authors are related to representatives of the genus Pseudomonas, 37% - to Xanthomonas and 19% - to Pectobacterium. PMID:23293826

  8. Septins Arrange F-Actin-Containing Fibers on the Chlamydia trachomatis Inclusion and Are Required for Normal Release of the Inclusion by Extrusion

    PubMed Central

    Volceanov, Larisa; Herbst, Katharina; Biniossek, Martin; Schilling, Oliver; Haller, Dirk; Nölke, Thilo; Subbarayal, Prema; Rudel, Thomas; Zieger, Barbara

    2014-01-01

    ABSTRACT Chlamydia trachomatis is an obligate intracellular human pathogen that grows inside a membranous, cytosolic vacuole termed an inclusion. Septins are a group of 13 GTP-binding proteins that assemble into oligomeric complexes and that can form higher-order filaments. We report here that the septins SEPT2, -9, -11, and probably -7 form fibrillar structures around the chlamydial inclusion. Colocalization studies suggest that these septins combine with F actin into fibers that encase the inclusion. Targeting the expression of individual septins by RNA interference (RNAi) prevented the formation of septin fibers as well as the recruitment of actin to the inclusion. At the end of the developmental cycle of C. trachomatis, newly formed, infectious elementary bodies are released, and this release occurs at least in part through the organized extrusion of intact inclusions. RNAi against SEPT9 or against the combination of SEPT2/7/9 substantially reduced the number of extrusions from a culture of infected HeLa cells. The data suggest that a higher-order structure of four septins is involved in the recruitment or stabilization of the actin coat around the chlamydial inclusion and that this actin recruitment by septins is instrumental for the coordinated egress of C. trachomatis from human cells. The organization of F actin around parasite-containing vacuoles may be a broader response mechanism of mammalian cells to the infection by intracellular, vacuole-dwelling pathogens. PMID:25293760

  9. Bacterial transformation of terpenoids

    NASA Astrophysics Data System (ADS)

    Grishko, V. V.; Nogovitsina, Y. M.; Ivshina, I. B.

    2014-04-01

    Data on the bacterial transformation of terpenoids published in the literature in the past decade are analyzed. Possible pathways for chemo-, regio- and stereoselective modifications of terpenoids are discussed. Considerable attention is given to new technological approaches to the synthesis of terpenoid derivatives suitable for the use in the perfume and food industry and promising as drugs and chiral intermediates for fine organic synthesis. The bibliography includes 246 references.

  10. Neglected Bacterial Zoonoses

    PubMed Central

    Chikeka, Ijeuru; Dumler, J. Stephen

    2015-01-01

    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. While 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 a broad spectrum diagnoses are actively sought. Thus, 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. PMID:25964152

  11. Acute Bacterial Cholangitis

    PubMed Central

    Zimmer, Vincent; Lammert, Frank

    2015-01-01

    Background Acute bacterial cholangitis for the most part owing to common bile duct stones is common in gastroenterology practice and represents a potentially life-threatening condition often characterized by fever, abdominal pain, and jaundice (Charcot's triad) as well as confusion and septic shock (Reynolds' pentad). Methods This review is based on a systematic literature review in PubMed with the search items ‘cholangitis’, ‘choledocholithiasis’, ‘gallstone disease’, ‘biliary infection’, and ‘biliary sepsis’. Results Although most patients respond to empiric broad-spectrum antibiotic treatment, timely endoscopic biliary drainage depending on the severity of the disease is required to eliminate the underlying obstruction. Specific recommendations have been derived from the Tokyo guideline working group consensus 2006 and its update in 2013, albeit poorly evidence-based, providing a comprehensive overview of diagnosis, classification, risk stratification, and treatment algorithms in acute bacterial cholangitis. Conclusion Prompt clinical recognition and accurate diagnostic workup including adequate laboratory assessment and (aetiology-oriented) imaging are critical steps in the management of cholangitis. Treatment is directed at the two major interrelated pathophysiologic components, i.e. bacterial infection (immediate antimicrobial therapy) and bile duct obstruction (biliary drainage). As for the latter, transpapillary endoscopic drainage by stent or nasobiliary drain and/or same-session bile duct clearance, depending on individual disease severity, represent first-line treatment approaches. PMID:26468310

  12. Cytoplasmic Inclusions of TDP-43 in Neurodegenerative Diseases: A Potential Role for Caspases

    PubMed Central

    Rohn, Troy T.

    2009-01-01

    TAR DNA-binding protein-43 (TDP-43) proteinopathies are classified based upon the extent of modified TDP-43 inclusions and include a growing number of neurodegenerative diseases including amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration with ubiquitin immunoreactive, tau negative inclusions (FTLD-U) and FTLD with motor neuron disease (FTLD-MND). In addition, TDP-43 inclusions have also been identified in a number of other neurodegenerative disorders including Alzheimer's disease, corticobasal degeneration, Lewy body related diseases and Pick's disease. Current understanding suggests that in these diseases, TDP-43 is relocated from the nucleus to the cytoplasm and sequestered into inclusions that contain modified TDP-43. Major modifications of TDP-43 have been identified as being hyperphosphorylation and proteolytic cleavage by caspases. In this review a summary of the major findings regarding the proteolytic modification of TDP-43 will be discussed as well as potential toxic-gain mechanisms these fragments may cause including cytoskeletal disruptions. PMID:19554515

  13. The Blue Angel. I - The mineralogy and petrogenesis of a hibonite inclusion from the Murchison meteorite

    NASA Astrophysics Data System (ADS)

    Armstrong, J. T.; Meeker, G. P.; Huneke, J. C.; Wasserburg, G. J.

    1982-04-01

    Results are presented of a mineralogical and petrological examination of a relatively large hibonite-containing inclusion in the Murchison chondrite called the Blue Angel. Mechanisms of formation and modification of hibonite-rich inclusions are investigated. Evidence is found that modifying processes occurred before the inclusion was incorporated into its present location, which indicates a sequence of aqueous and metamorphic alteration followed by disruption and relithification of the protometeoritic material. The mineral chemistry and petrography of the Blue Angel is consistent with a three stage formation history, and extensive alteration of Ca-Al-rich inclusions may have occurred by aqueous alteration and thermal metamorphism followed by explosive mixing processes on a parent body.

  14. In vivo deuteration strategies for neutron scattering analysis of bacterial polyhydroxyoctanoate.

    PubMed

    Russell, Robert A; Holden, Peter J; Wilde, Karyn L; Garvey, Christopher J; Hammerton, Kerie M; Foster, L John R

    2008-06-01

    The cultivation of microorganisms on deuterated substrates has allowed us to control deuterium incorporation into biopolymer systems which is important for characterisation using neutron scattering techniques. Bacterial polyhydroxyoctanoate (PHO) is a polyester formed within inclusions inside bacterial cells and was deuterated in vivo under various conditions to characterise the formation of these inclusions by neutron scattering. Manipulation of deuterated media during microbial growth and PHO production phases resulted in polymer with partial or complete substitution of hydrogen by deuterium, as shown by gas chromatography. Sequential feeding of hydrogenated and deuterated forms of the same precursor was used to demonstrate that neutron scattering analysis could be used to differentiate between chemically similar phases in these polymer inclusions. PMID:18481053

  15. Production of stabilized scFv antibody fragments in the E. coli bacterial cytoplasm.

    PubMed

    Vaks, Lilach; Benhar, Itai

    2014-01-01

    Monoclonal antibodies (mAbs) are currently the fastest growing class of therapeutic proteins. Parallel to full-length IgG format the development of recombinant technologies provided the production of smaller recombinant antibody variants. The single-chain variable fragment (scFv) antibody is a minimal form of functional antibody comprised of the variable domains of immunoglobulin light and heavy chains connected by a flexible linker. In most cases, scFvs are expressed in the bacterium E. coli. The production of soluble scFvs under the reducing conditions of the E. coli bacterial cytoplasm is inefficient because of the inability of the disulfide bonds to form. Hence, scFvs are either secreted to the periplasm as soluble proteins or expressed in the cytoplasm as insoluble inclusion bodies and recovered by refolding. The cytoplasmic expression of scFvs as a C-terminal fusion to maltose-binding protein (MBP) provided the high-level production of stable, soluble, and functional fusion protein. The below protocol provides the detailed description of MBP-scFv production in E. coli utilizing two expression systems: pMalc-TNN and pMalc-NHNN. Although the MBP tag does not disrupt the most of antibody activities, the MBP-TNN-scFv product can be cleaved by TEV protease in order to obtain untagged scFv. PMID:24037842

  16. Supporting Students with Severe Disabilities in Inclusive Schools: A Descriptive Account From Schools Implementing Inclusive Practices

    ERIC Educational Resources Information Center

    Kurth, Jennifer A.; Lyon, Kristin J.; Shogren, Karrie A.

    2015-01-01

    The purpose of the present study was to investigate practices that support the inclusion of students with severe disabilities in the learning and social activities of inclusive K-8 schools to inform inclusive school reform research and practice. Eighteen K-8 students with severe disabilities in six schools recognized for their implementation of…

  17. Full Inclusion: The Benefits and Disadvantages of Inclusive Schooling. An Overview

    ERIC Educational Resources Information Center

    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…

  18. More Policies, Greater Inclusion? Exploring the Contradictions of New Labour Inclusive Education Policy

    ERIC Educational Resources Information Center

    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…

  19. Measuring the Quality of Inclusive Practices: Findings from the Inclusive Classroom Profile Pilot

    ERIC Educational Resources Information Center

    Soukakou, Elena P.; Winton, Pam J.; West, Tracey A.; Sideris, John H.; Rucker, Lia M.

    2014-01-01

    The purpose of this study was to test the reliability and validity of the Inclusive Classroom Profile (ICP), an observation measure designed to assess the quality of classroom practices in inclusive preschool programs. The measure was field tested in 51 inclusive classrooms. Results confirmed and extended previous research findings, providing…

  20. What's Working: Topics in Inclusive Education.

    ERIC Educational Resources Information Center

    What's Working, 1994

    1994-01-01

    This newsletter focuses on what works in inclusive education. A major article is "Inclusive Education: Needs of Minnesota Families--Major Findings from a 'Together We're Better' Study." This article presents findings of a statewide survey of families of children with disabilities. Findings are categorized within four areas: (1) educational…

  1. How Does Supervision Support Inclusive Teacherhood?

    ERIC Educational Resources Information Center

    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…

  2. Early Childhood Inclusion: Focus on Change.

    ERIC Educational Resources Information Center

    Guralnick, Michael J., Ed.

    Early childhood inclusion is a field characterized by a philosophy and practice that encourages full participation of children with disabilities and their families in everyday activities with their typically developing peers. Noting that success in inclusion endeavors requires substantial changes in the way our society thinks, feels, and acts,…

  3. Metaphors of Literacy: Dialogues in Inclusive Settings

    ERIC Educational Resources Information Center

    Causarano, Antonio

    2015-01-01

    This article discusses the importance of metaphors in education and in inclusive settings in particular. Metaphors are seen as the fabric of collaboration through dialogue across the curriculum. The article analyzes the dialogues among the Language Arts, Social Studies, and inclusion teacher in a large middle school in the Southwest of the United…

  4. Inclusive Focus Particles in English and Korean

    ERIC Educational Resources Information Center

    Kang, Sang-gu

    2011-01-01

    When discussing focus particles, it has been common practice to rely on the dichotomy of inclusive vs. exclusive particles, "a la" Konig (1991). Inclusive focus particles are often further divided into scalar particles, such as "also", "too", and "either", and non-scalar particles, such as "even". In this thesis, I advance a comparative analysis…

  5. Inclusive Education in Malaysia: Policy and Practice

    ERIC Educational Resources Information Center

    Jelas, Zalizan M.; Ali, Manisah Mohd

    2014-01-01

    Malaysia's move towards inclusion was given impetus by its participation in workshops and conferences set up under the auspices of the United Nations (UNESCO 1990; UN 1993; UNESCO 1994). Inclusive education was introduced in the Education Act 1996 as part of the continuum of services available for children with special needs. The purpose of…

  6. Inclusive Education: Teacher Perspectives from Malaysia

    ERIC Educational Resources Information Center

    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…

  7. International Perspectives on Inclusion: Concluding Thoughts.

    ERIC Educational Resources Information Center

    Timmons, Vianne

    2002-01-01

    This article discusses overall trends and commonalities among previous articles on inclusion in the United Kingdom, Ireland, India, South Africa, Canada, New Zealand, and Australia. It concludes that inclusive education must be viewed in a broader context that looks at school reform and the societal context. Teacher education is stressed.…

  8. Social Inclusion and Critical Consciousness in Australia

    ERIC Educational Resources Information Center

    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…

  9. The Challenges of Inclusive Education in Lebanon

    ERIC Educational Resources Information Center

    Wehbi, Samantha

    2006-01-01

    This article reports on a study conducted in Lebanon by four non-governmental organizations, primarily on the topic of inclusion of people with disabilities in education, with a secondary focus on inclusion in employment. This article reports on some of the findings of the assessment process that was four months in duration and aimed, through…

  10. The Sustainability of Inclusive School Reform

    ERIC Educational Resources Information Center

    Sindelar, Paul T.; Shearer, Deirdre K.; Yendol-Hoppey, Diane; Liebert, Todd W.

    2006-01-01

    For over a decade, University of Florida researchers worked with middle schools in a large urban and suburban south Florida district, as they developed and then worked to sustain inclusive reform. One middle school, Socrates, was notably successful, having built its inclusion model on a foundation of previous reform and a school culture…

  11. Geometric Modeling of Inclusions as Ellipsoids

    NASA Technical Reports Server (NTRS)

    Bonacuse, Peter J.

    2008-01-01

    Nonmetallic inclusions in gas turbine disk alloys can have a significant detrimental impact on fatigue life. Because large inclusions that lead to anomalously low lives occur infrequently, probabilistic approaches can be utilized to avoid the excessively conservative assumption of lifing to a large inclusion in a high stress location. A prerequisite to modeling the impact of inclusions on the fatigue life distribution is a characterization of the inclusion occurrence rate and size distribution. To help facilitate this process, a geometric simulation of the inclusions was devised. To make the simulation problem tractable, the irregularly sized and shaped inclusions were modeled as arbitrarily oriented, three independent dimensioned, ellipsoids. Random orientation of the ellipsoid is accomplished through a series of three orthogonal rotations of axes. In this report, a set of mathematical models for the following parameters are described: the intercepted area of a randomly sectioned ellipsoid, the dimensions and orientation of the intercepted ellipse, the area of a randomly oriented sectioned ellipse, the depth and width of a randomly oriented sectioned ellipse, and the projected area of a randomly oriented ellipsoid. These parameters are necessary to determine an inclusion s potential to develop a propagating fatigue crack. Without these mathematical models, computationally expensive search algorithms would be required to compute these parameters.

  12. Culturally Inclusive Curriculum in Higher Education

    ERIC Educational Resources Information Center

    Pridham, Bruce; Martin, Dona; Walker, Kym; Rosengren, Rosie; Wadley, Danielle

    2015-01-01

    The Australian National Program Standards for Teacher Education prioritises knowledge of culturally inclusive practices and challenges the educational community to present research on well-structured, inclusive, cross-curricula education partnerships. This article meets this challenge as it explores a core unit of work for undergraduate teachers…

  13. Standards & Inclusion: Can We Have Both?. [Videotape].

    ERIC Educational Resources Information Center

    Lipsky, Dorothy Kerzner; Gartner, Alan

    Designed for regular and special educators, this 40-minute video profiles inclusive schools from around the country that have successfully incorporated academic standards. It addresses many of the critical issues facing educators who are supporting students with disabilities in inclusive settings. Topics discussed include: (1) the consequences of…

  14. Questions of Inclusion in Scotland and Europe

    ERIC Educational Resources Information Center

    Allan, Julie

    2010-01-01

    This paper examines inclusion in Scotland and in Europe. It considers some of the uncertainties surrounding inclusion and the questions--many of which give cause for concern--that are currently being raised by researchers, teachers and their representative unions, parents and children. The shifting political and policy contexts and recent patterns…

  15. Inclusive Education: A Series of Issue Papers.

    ERIC Educational Resources Information Center

    Illinois Coalition on School Inclusion, Springfield.

    Thirteen brief papers address issues in inclusive education, with special emphasis on concerns and applications in Illinois. The following issues are discussed: (1) the relationship of the inclusive schools concept to that of providing a continuum of alternative placement options; (2) needed changes in current funding of special education to…

  16. Inclusion: Something More Than Sitting Together

    ERIC Educational Resources Information Center

    Bulbul, Mustafa Sahin

    2011-01-01

    Awareness about students with learning difficulty brought us to inclusive learning environments. The acceptance was to build collaborative atmospheres in the class. Unfortunately, when teachers are not enthusiastic and adequate to develop the interaction, the inclusive learning environment never occurs. This conclusion bases on this studies…

  17. Classroom Groups: Inclusion of Students with Disabilities.

    ERIC Educational Resources Information Center

    Fields, Teesue H.

    1995-01-01

    Discusses the inclusion of students with disabilities in the regular classroom group. Analyzes the structure of the classroom group to determine its receptivity to students with disabilities. Presents special problems that might be encountered during inclusion and possibilities for improving classroom group skills. (LKS)

  18. Case Studies for Inclusive Schools. Second Edition

    ERIC Educational Resources Information Center

    Anderson, Peggy L.

    2005-01-01

    Case Studies for "Inclusive Schools, Second Edition" presents a sampling of case studies that contain realistic problems concerning inclusion issues for teacher education students to solve. This format was chosen because the case study approach to learning is gaining in popularity as it provides students with an opportunity to apply information…

  19. Design, Development and Implementation of Inclusive Education

    ERIC Educational Resources Information Center

    Mooij, Ton; Smeets, Ed

    2006-01-01

    In inclusive education different pupils, including pupils with special educational needs and high ability pupils, can be stimulated to learn according to their capacities and potentials. The research question concentrates on the design features of inclusive education that will optimally promote the motivation and learning processes and outcomes of…

  20. Erasmus Mundus SEN: The Inclusive Scholarship Programme?

    ERIC Educational Resources Information Center

    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,…

  1. Tensions around Inclusion: Reframing the Moral Horizon

    ERIC Educational Resources Information Center

    Clegg, Jennifer; Murphy, Elizabeth; Almack, Kathryn; Harvey, Anna

    2008-01-01

    Background: Inclusion is one of four policies for people with disabilities in the UK. Criticisms of its three key attributes--mainstreaming, independent living and employment--are reviewed. Methods: This study of 28 young people, most with severe intellectual disabilities, investigated engagement with inclusion at their transition to adult…

  2. Inclusive Partnership: Enhancing Student Engagement in Geography

    ERIC Educational Resources Information Center

    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…

  3. Beyond Political Correctness: Toward the Inclusive University.

    ERIC Educational Resources Information Center

    Richer, Stephen, Ed.; Weir, Lorna, Ed.

    This collection of 12 essays examines the history of the discourse over political correctness (PC) in Canadian academia, focusing on the neoconservative backlash to affirmative action, inclusive policies, and feminist and anti-racist teaching in the classroom. It includes: (1) "Introduction: Political Correctness and the Inclusive University"…

  4. Inclusive Education: A Servant-Leadership Perspective

    ERIC Educational Resources Information Center

    Crippen, Carolyn

    2005-01-01

    The concept of inclusive education is common to the Canadian educational system and involves service to the students. According to Young and Levin, "The provision of appropriate education to meet the needs of each student is increasingly accepted as a right in Canadian education." Bloom, Perlmutter and Burnell define inclusion as "a philosophy…

  5. Older Women's Career Development and Social Inclusion

    ERIC Educational Resources Information Center

    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…

  6. The Inclusive Secondary School Teacher in Australia

    ERIC Educational Resources Information Center

    Pearce, Michelle

    2009-01-01

    As a result of the 1992 Disability Discrimination Act in Australia and parental support for inclusion, regular teachers now include students with disabilities in their classes. Inclusion has been more successful in primary than in secondary schools. Secondary schools remain a challenge due to their traditional focus on curriculum, examinations,…

  7. Irish Mathematics Teachers' Attitudes towards Inclusion

    ERIC Educational Resources Information Center

    Whitty, Elaine; Clarke, Marie

    2012-01-01

    This paper through the theoretical framework of constructive attitude theory explores mathematics teachers' attitudes and pedagogical strategies with reference to inclusive practice. The authors argue that though teachers may have formed positive inclusive attitudes, the translation of these into practice does not always occur and poses…

  8. Characteristics of Inclusive Classrooms in Turkey

    ERIC Educational Resources Information Center

    Melekoglu, Macid Ayhan

    2014-01-01

    In this paper, inclusive classrooms in Turkey are described in terms of the characteristics of both classroom teachers and students with special needs. Participants of this study consisted of 54 teachers working in inclusive classrooms and 54 students with mild intellectual disabilities in those classrooms in Turkey. Data for this study were…

  9. Enhancing Theory Courses with Racially Inclusive Research.

    ERIC Educational Resources Information Center

    Bramlett-Solomon, Sharon; Liebler, Carol M.

    1999-01-01

    Offers a blueprint that instructors of mass media theory courses can adopt to expose students to racially inclusive research in order to encourage students to explore and employ relevant theories when probing media and race questions. Offers examples of inclusive media research, examining six prominent theories: selective-perception theory,…

  10. Students' Perspectives on LGBTQ-Inclusive Curriculum

    ERIC Educational Resources Information Center

    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…

  11. Moving Forward in Inclusive Education Research

    ERIC Educational Resources Information Center

    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…

  12. Inclusive Education in Thailand: Practices and Challenges

    ERIC Educational Resources Information Center

    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…

  13. Beyond Prejudice: Inclusive Learning in Practice

    ERIC Educational Resources Information Center

    Smith, Vikki; Armstrong, Anne

    2005-01-01

    Promoting an inclusive learning environment that caters for all learners and their individual needs and meeting challenging targets set in this area is a huge under taking for providers across the learning and skills sector. This booklet provides an overview that illustrates the breadth and variety that the broad banner of inclusive learning…

  14. Inclusive Schooling Policy: An Educational Detective Story?

    ERIC Educational Resources Information Center

    Moss, Julianne

    2003-01-01

    Since the publication of the Salamanca statement (UNESCO 1994), inclusive schooling has formed a growing part of the deliberations of the special education community. Inclusive schooling research in Australia in the main continues to reproduce traditions of the special education field, emphasising the dominant psychological perspectives that have…

  15. Inclusive Education: Is This Horse a Trojan?

    ERIC Educational Resources Information Center

    Slee, Roger

    2006-01-01

    In Canada and elsewhere, governments are expending considerable effort in the production of inclusive education policy texts, resources allocation models, and programs. The author notes that despite of the analytic power and the political intent of inclusive education as a counterpoint to special education, its appropriation is imminent if not…

  16. Secondary General Education Teachers' Attitudes toward Inclusion

    ERIC Educational Resources Information Center

    Wilson, Valerie A.

    2014-01-01

    Since the inception of the Individuals with Disabilities Education Improvement Act, educators have struggled with including students with disabilities in the general education classroom with their nondisabled peers. The inclusion educational model was utilized in this study to explore secondary teachers' attitudes toward inclusive educational…

  17. Striving for Quality in Early Childhood Inclusion

    ERIC Educational Resources Information Center

    Brancato, Kimberly

    2013-01-01

    An essential component of best practice in the field of early childhood special education is the inclusion of children with disabilities in typical early childhood settings. As the practice of inclusion has increased in recent years it has become imperative to ensure that children with disabilities attend quality programs. The main purpose of this…

  18. Teachers' Misunderstanding: The Concept of Inclusive Education

    ERIC Educational Resources Information Center

    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…

  19. International Teachers' Attitudes towards Inclusion in Turkey

    ERIC Educational Resources Information Center

    Pursley, Lisa

    2014-01-01

    The inclusion of students with disabilities in the general education classroom has become a universally debated topic in education. More schools all over the world are seeking to use inclusion as their main service delivery model for students with disabilities. Much research has been conducted globally to gain insights into general education…

  20. Guided Reading in Inclusive Middle Years Classrooms

    ERIC Educational Resources Information Center

    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…