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Sample records for heme prosthetic group

  1. Peroxide-Dependent Analyte Conversion by the Heme Prosthetic Group, the Heme Peptide “Microperoxidase-11” and Cytochrome c on Chitosan Capped Gold Nanoparticles Modified Electrodes

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    Frieder W. Scheller

    2012-05-01

    Full Text Available In view of the role ascribed to the peroxidatic activity of degradation products of cytochrome c (cyt c in the processes of apoptosis, we investigate the catalytic potential of heme and of the cyt c derived heme peptide MP-11 to catalyse the cathodic reduction of hydrogen peroxide and to oxidize aromatic compounds. In order to check whether cyt c has an enzymatic activity in the native state where the protein matrix should suppress the inherent peroxidatic activity of its heme prosthetic group, we applied a biocompatible immobilization matrix and very low concentrations of the co-substrate H2O2. The biocatalysts were entrapped on the surface of a glassy carbon electrode in a biocompatible chitosan layer which contained gold nanoparticles. The electrochemical signal for the peroxide reduction is generated by the redox conversion of the heme group, whilst a reaction product of the substrate oxidation is cathodically reduced in the substrate indication. The catalytic efficiency of microperoxidase-11 is sufficient for sensors indicating HRP substrates, e.g., p-aminophenol, paracetamol and catechol, but also the hydroxylation of aniline and dehalogenation of 4-fluoroaniline. The lower limit of detection for p-aminophenol is comparable to previously published papers with different enzyme systems. The peroxidatic activity of cyt c immobilized in the chitosan layer for catechol was found to be below 1 per mill and for p-aminophenol about 3% as compared with that of heme or MP-11.

  2. Environmental heme utilization by heme-auxotrophic bacteria.

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    Gruss, Alexandra; Borezée-Durant, Elise; Lechardeur, Delphine

    2012-01-01

    Heme, an iron-containing porphyrin, is the prosthetic group for numerous key cellular enzymatic and regulatory processes. Many bacteria encode the biosynthetic enzymes needed for autonomous heme production. Remarkably, however, numerous other bacteria lack a complete heme biosynthesis pathway, yet encode heme-requiring functions. For such heme-auxotrophic bacteria (HAB), heme or porphyrins must be captured from the environment. Functional studies, aided by genomic analyses, provide insight into the HAB lifestyle, how they acquire and manage heme, and the uses of heme that make it worthwhile, and sometimes necessary, to capture this bioactive molecule. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. Heme Gazing: Illuminating Eukaryotic Heme Trafficking, Dynamics, and Signaling with Fluorescent Heme Sensors.

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    Hanna, David A; Martinez-Guzman, Osiris; Reddi, Amit R

    2017-04-04

    Heme (iron protoporphyrin IX) is an essential protein prosthetic group and signaling molecule required for most life on Earth. All heme-dependent processes require the dynamic and rapid mobilization of heme from sites of synthesis or uptake to hemoproteins present in virtually every subcellular compartment. The cytotoxicity and hydrophobicity of heme necessitate that heme mobilization be carefully controlled to mitigate the deleterious effects of this essential toxin. Indeed, a number of disorders, including certain cancers, cardiovascular diseases, and aging and age-related neurodegenerative diseases, are tied to defects in heme homeostasis. However, the molecules and mechanisms that mediate heme transport and trafficking, and the dynamics of these processes, are poorly understood. This is in large part due to the lack of physical tools for probing cellular heme. Herein, we discuss the recent development of fluorescent probes that can monitor and image kinetically labile heme with respect to its mobilization and role in signaling. In particular, we will highlight how heme gazing with these tools can uncover new heme trafficking factors upon being integrated with genetic screens and illuminate the concentration, subcellular distribution, and dynamics of labile heme in various physiological contexts. Altogether, the monitoring of labile heme, along with recent biochemical and cell biological studies demonstrating the reversible regulation of certain cellular processes by heme, is challenging us to reconceptualize heme from being a static cofactor buried in protein active sites to a dynamic and mobile signaling molecule.

  4. Kinetics of heme transfer by the Shr NEAT domains of Group A Streptococcus.

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    Ouattara, Mahamoudou; Pennati, Andrea; Devlin, Darius J; Huang, Ya-Shu; Gadda, Giovanni; Eichenbaum, Zehava

    2013-10-15

    The hemolytic Group A Streptococcus (GAS) is a notorious human pathogen. Shr protein of GAS participates in iron acquisition by obtaining heme from host hemoglobin and delivering it to the adjacent receptor on the surface, Shp. Heme is then conveyed to the SiaABC proteins for transport across the membrane. Using rapid kinetic studies, we investigated the role of the two heme binding NEAT modules of Shr. Stopped-flow analysis showed that holoNEAT1 quickly delivered heme to apoShp. HoloNEAT2 did not exhibit such activity; only little and slow transfer of heme from NEAT2 to apoShp was seen, suggesting that Shr NEAT domains have distinctive roles in heme transport. HoloNEAT1 also provided heme to apoNEAT2, by a fast and reversible process. To the best of our knowledge this is the first transfer observed between isolated NEAT domains of the same receptor. Sequence alignment revealed that Shr NEAT domains belong to two families of NEAT domains that are conserved in Shr orthologs from several species. Based on the heme transfer kinetics, we propose that Shr proteins modulate heme uptake according to heme availability by a mechanism where NEAT1 facilitates fast heme delivery to Shp, whereas NEAT2 serves as a temporary storage for heme on the bacterial surface. Copyright © 2013. Published by Elsevier Inc.

  5. The use and abuse of heme in apicomplexan parasites.

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    van Dooren, Giel G; Kennedy, Alexander T; McFadden, Geoffrey I

    2012-08-15

    Heme is an essential prosthetic group for most life on Earth. It functions in numerous cellular redox reactions, including in antioxidant defenses and at several stages of the electron transport chain in prokaryotes and eukaryotic mitochondria. Heme also functions as a sensor and transport molecule for gases such as oxygen. Heme is a complex organic molecule and can only be synthesized through a multienzyme pathway from simpler precursors. Most free-living organisms synthesize their own heme by a broadly conserved metabolic pathway. Parasites are adept at scavenging molecules from their hosts, and heme is no exception. In this review we examine recent advances in understanding heme usage and acquisition in Apicomplexa, a group of parasites that include the causative agents of malaria, toxoplasmosis, and several major parasites of livestock. Heme is critical to the survival of Apicomplexa, although the functions of heme in these organisms remain poorly understood. Some Apicomplexa likely scavenge heme from their host organisms, while others retain the ability to synthesize heme. Surprisingly, some Apicomplexa may be able to both synthesize and scavenge heme. Several Apicomplexa live in intracellular environments that contain high levels of heme. Since heme is toxic at high concentrations, parasites must carefully regulate intracellular heme levels and develop mechanisms to detoxify excess heme. Indeed, drugs interfering with heme detoxification serve as major antimalarials. Understanding heme requirements and regulation in apicomplexan parasites promises to reveal multiple targets for much-needed therapeutic intervention against these parasites.

  6. Corynebacterium CDC group G native and prosthetic valve endocarditis

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    Adil Sattar

    2015-08-01

    Full Text Available We report the first case of native and recurrent prosthetic valve endocarditis with Corynebacterium CDC group G, a rarely reported cause of infective endocarditis (IE. Previously, there have been only two cases reported for prosthetic valve IE caused by these organisms. A 69-year-old female with a known history of mitral valve regurgitation presented with a 3-day history of high-grade fever, pleuritic chest pain and cough. Echocardiography confirmed findings of mitral valve thickening consistent with endocarditis, which subsequently progressed to become large and mobile vegetations. Both sets of blood cultures taken on admission were positive for Corynebacterium CDC group G. Despite removal of a long-term venous access port, the patient’s presumed source of line associated bacteremia, mitral valve replacement, and aggressive antibiotic therapy, the patient had recurrence of vegetations on the prosthetic valve. She underwent replacement of her prosthetic mitral valve in the subsequent 2 weeks, before she progressed to disseminated intravascular coagulation and expired. Although they are typically considered contaminants, corynebacteria, in the appropriate clinical setting, should be recognized, identified, and treated as potentially life-threatening infections, particularly in the case of line-associated bacteremias, and native and prosthetic valve endocarditis.

  7. Corynebacterium CDC Group G Native and Prosthetic Valve Endocarditis.

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    Sattar, Adil; Yu, Siegfried; Koirala, Janak

    2015-08-11

    We report the first case of native and recurrent prosthetic valve endocarditis with Corynebacterium CDC group G, a rarely reported cause of infective endocarditis (IE). Previously, there have been only two cases reported for prosthetic valve IE caused by these organisms. A 69-year-old female with a known history of mitral valve regurgitation presented with a 3-day history of high-grade fever, pleuritic chest pain and cough. Echocardiography confirmed findings of mitral valve thickening consistent with endocarditis, which subsequently progressed to become large and mobile vegetations. Both sets of blood cultures taken on admission were positive for Corynebacterium CDC group G. Despite removal of a long-term venous access port, the patient's presumed source of line associated bacteremia, mitral valve replacement, and aggressive antibiotic therapy, the patient had recurrence of vegetations on the prosthetic valve. She underwent replacement of her prosthetic mitral valve in the subsequent 2 weeks, before she progressed to disseminated intravascular coagulation and expired. Although they are typically considered contaminants, corynebacteria, in the appropriate clinical setting, should be recognized, identified, and treated as potentially life-threatening infections, particularly in the case of line-associated bacteremias, and native and prosthetic valve endocarditis.

  8. A New F-18 Prosthetic Group via an Oxime Coupling

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    Carberry, Patrick; Lieberman, Brian P.; Ploessl, Karl; Choi, Seok R.; Haase, Danniebelle N.; Kung, Hank F.

    2011-01-01

    A novel fluorine-18 prosthetic ligand, 5-(1,3-dioxolan-2-yl)-2-(2-(2-(2- fluoroethoxy)ethoxy)ethoxy)pyridine [18F]2, has been synthesized. The prosthetic ligand is formed in high radiochemical yield (rcy = 71 ± 2 %, n = 3) with excellent radiochemical purity (rcp = 99 ± 1 %, n = 3) in a short reaction time (10 min). [18F]2 is a small, neutral, organic complex, easily synthesized in four steps from a readily available starting material. It can be anchored onto a target molecule containing an aminooxy functional group under acidic conditions by way of an oxime bond. We report herein two examples [18F]23 and [18F]24, potential imaging agents for β-amyloid plaques, which were labeled with this prosthetic group. This approach could be used for labeling proteins and peptides containing an aminooxy group. Biodistribution in male ICR mice for both oxime labeled complexes [18F]23 and [18F]24 were compared to that of the known β-amyloid plaque indicator, [18F]-AV-45, florbetapir 1. Oximes [18F]23 and [18F]24 are larger in size and therefore should reduce the blood-brain barrier (BBB) penetration. The brain uptake for oxime [18F]23 appeared to be reduced, but still retained some capability to cross the BBB. Oxime [18F]24 showed promising results after 2 min post injection (0.48 % dose/gram), however the uptake increased after 30 min post injection (0.92 % dose/gram) suggesting an in-vivo decomposition/metabolism of compound [18F]24. We have demonstrated a general protocol for the fluoride-18 labeling with a new prosthetic ligand [18F]2 that is tolerant towards several functional groups and is formed via chemoselective oxime coupling. PMID:21452846

  9. Biology of Heme in Mammalian Erythroid Cells and Related Disorders

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    Tohru Fujiwara

    2015-01-01

    Full Text Available Heme is a prosthetic group comprising ferrous iron (Fe2+ and protoporphyrin IX and is an essential cofactor in various biological processes such as oxygen transport (hemoglobin and storage (myoglobin and electron transfer (respiratory cytochromes in addition to its role as a structural component of hemoproteins. Heme biosynthesis is induced during erythroid differentiation and is coordinated with the expression of genes involved in globin formation and iron acquisition/transport. However, erythroid and nonerythroid cells exhibit distinct differences in the heme biosynthetic pathway regulation. Defects of heme biosynthesis in developing erythroblasts can have profound medical implications, as represented by sideroblastic anemia. This review will focus on the biology of heme in mammalian erythroid cells, including the heme biosynthetic pathway as well as the regulatory role of heme and human disorders that arise from defective heme synthesis.

  10. Biology of Heme in Mammalian Erythroid Cells and Related Disorders

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    Fujiwara, Tohru; Harigae, Hideo

    2015-01-01

    Heme is a prosthetic group comprising ferrous iron (Fe2+) and protoporphyrin IX and is an essential cofactor in various biological processes such as oxygen transport (hemoglobin) and storage (myoglobin) and electron transfer (respiratory cytochromes) in addition to its role as a structural component of hemoproteins. Heme biosynthesis is induced during erythroid differentiation and is coordinated with the expression of genes involved in globin formation and iron acquisition/transport. However, erythroid and nonerythroid cells exhibit distinct differences in the heme biosynthetic pathway regulation. Defects of heme biosynthesis in developing erythroblasts can have profound medical implications, as represented by sideroblastic anemia. This review will focus on the biology of heme in mammalian erythroid cells, including the heme biosynthetic pathway as well as the regulatory role of heme and human disorders that arise from defective heme synthesis. PMID:26557657

  11. Models for cytochrome P450 prosthetic heme alkylation. Reaction of diazoacetophenone with (tetraphenylporphyrinato)iron(II) chloride

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    Komives, E.A.; Tew, D.; Olmstead, M.M.; Ortiz de Montellano, P.R.

    1988-09-07

    The reaction of diazoacetophenone with (tetraphenylporphyrinato)iron(II) yields (N-(2-phenyl-2-oxoethyl)tetraphenylporphyrinato)iron(II) chloride. The structure of this product has been established by spectroscopic methods and by x-ray crystallography. The crystal structure shows that the first carbon of the N-alkyl group is 2.94 /angstrom/ from the iron atom and that the oxygen of the N-alkyl group points away from the iron. No evidence is seen for the Fe-C-N product expected from insertion of the diazo carbon into the metalloporphyrin iron-nitrogen bond or for intermediates in which the oxygen of the N-(2-phenyl-2-oxoethyl) group is coordinated to the iron. These results suggest it is unlikely that carbene-insertion or oxygen-coordinated intermediates will be detected during the N-alkylation of cytochrome P450 by diazo ketones. The results also rationalize the failure to detect iron-chelated enol species during N-alkylation of the prosthetic group of cytochrome P450 by catalytically activated phenylacetylene. 43 references, 5 figures, 4 tables.

  12. Heme metabolism in stress regulation and protein production: From Cinderella to a key player.

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    Martínez, J L; Petranovic, D; Nielsen, J

    2016-04-02

    Heme biosynthesis is a highly conserved pathway which is present in all kingdoms, from Archaea to higher organisms such as plants and mammals. The heme molecule acts as a prosthetic group for different proteins and enzymes involved in energy metabolism and reactions involved in electron transfer. Based on our recent findings and other recent reports, we here illustrate that heme is more than a co-factor. We also discuss the necessity to gain more insight into the heme biosynthesis pathway regulation, as this interacts closely with overall stress control. Understanding heme biosynthesis and its regulation could impact our ability to develop more efficient yeast cell factories for heterologous protein production.

  13. Heme metabolism in stress regulation and protein production: from Cinderella to a key player

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    Martinez Ruiz, José Luis; Petranovic, D.; Nielsen, Jens

    2016-01-01

    Heme biosynthesis is a highly conserved pathway which is present in all kingdoms, from Archaea to higher organisms such as plants and mammals. The heme molecule acts as a prosthetic group for different proteins and enzymes involved in energy metabolism and reactions involved in electron transfer....

  14. Heme and erythropoieis: more than a structural role

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    Chiabrando, Deborah; Mercurio, Sonia; Tolosano, Emanuela

    2014-01-01

    Erythropoiesis is the biological process that consumes the highest amount of body iron for heme synthesis. Heme synthesis in erythroid cells is finely coordinated with that of alpha (α) and beta (β)-globin, resulting in the production of hemoglobin, a tetramer of 2α- and 2β-globin chains, and heme as the prosthetic group. Heme is not only the structural component of hemoglobin, but it plays multiple regulatory roles during the differentiation of erythroid precursors since it controls its own synthesis and regulates the expression of several erythroid-specific genes. Heme is synthesized in developing erythroid progenitors by the stage of proerythroblast, through a series of eight enzymatic reactions divided between mitochondria and cytosol. Defects of heme synthesis in the erythroid lineage result in sideroblastic anemias, characterized by microcytic anemia associated to mitochondrial iron overload, or in erythropoietic porphyrias, characterized by porphyrin deposition in erythroid cells. Here, we focus on the heme biosynthetic pathway and on human erythroid disorders due to defective heme synthesis. The regulatory role of heme during erythroid differentiation is discussed as well as the heme-mediated regulatory mechanisms that allow the orchestration of the adaptive cell response to heme deficiency. PMID:24881043

  15. Characterizing mobility from the prosthetic limb user's perspective: Use of focus groups to guide development of the Prosthetic Limb Users Survey of Mobility.

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    Hafner, Brian J; Morgan, Sara J; Abrahamson, Daniel C; Amtmann, Dagmar

    2016-10-01

    Input from target respondents in the development of patient-reported outcome measures is necessary to ensure that the instrument is meaningful. To solicit perspectives of prosthetic limb users about their mobility experiences and to inform development of the Prosthetic Limb Users Survey of Mobility. Qualitative study. Four focus groups of lower limb prosthesis users were held in different regions of the United States. Focus group transcripts were coded, and themes were identified. Feedback from participants was used to develop a framework for measuring mobility with a lower limb prosthesis. Focus group participants (N = 37) described mobility as a confluence of factors that included characteristics of the individual, activity, and environment. Identified themes were defined as individual characteristics, forms of movement, and environmental situations. Prosthetic mobility was conceptualized as movement activities performed in an environmental or situational context. Respondent feedback used to guide development of Prosthetic Limb Users Survey of Mobility established a foundation for a new person-centered measure of mobility with a prosthetic limb. Perspectives of target respondents are needed to guide development of instruments intended to measure health outcomes. Focus groups of prosthetic limb users were conducted to solicit experiences related to mobility with a lower limb prosthesis. Results were used to inform development of a clinically meaningful, person-centered instrument. © The International Society for Prosthetics and Orthotics 2015.

  16. Molecular hijacking of siroheme for the synthesis of heme and d1 heme

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    Bali, Shilpa; Lawrence, Andrew D.; Lobo, Susana A.; Saraiva, Lígia M.; Golding, Bernard T.; Palmer, David J.; Howard, Mark J.; Ferguson, Stuart J.; Warren, Martin J.

    2011-01-01

    Modified tetrapyrroles such as chlorophyll, heme, siroheme, vitamin B12, coenzyme F430, and heme d1 underpin a wide range of essential biological functions in all domains of life, and it is therefore surprising that the syntheses of many of these life pigments remain poorly understood. It is known that the construction of the central molecular framework of modified tetrapyrroles is mediated via a common, core pathway. Herein a further branch of the modified tetrapyrrole biosynthesis pathway is described in denitrifying and sulfate-reducing bacteria as well as the Archaea. This process entails the hijacking of siroheme, the prosthetic group of sulfite and nitrite reductase, and its processing into heme and d1 heme. The initial step in these transformations involves the decarboxylation of siroheme to give didecarboxysiroheme. For d1 heme synthesis this intermediate has to undergo the replacement of two propionate side chains with oxygen functionalities and the introduction of a double bond into a further peripheral side chain. For heme synthesis didecarboxysiroheme is converted into Fe-coproporphyrin by oxidative loss of two acetic acid side chains. Fe-coproporphyrin is then transformed into heme by the oxidative decarboxylation of two propionate side chains. The mechanisms of these reactions are discussed and the evolutionary significance of another role for siroheme is examined. PMID:21969545

  17. Shr of Group A Streptococcus is a new type of composite NEAT protein involved in sequestering heme from methemoglobin

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    Ouattara, Mahamoudou; Cunha, Elizabeth Bentley; Li, Xueru; Huang, Ya-Shu; Dixon, Dabney; Eichenbaum, Zehava

    2010-01-01

    SUMMARY A growing body of evidence suggests that surface or secreted proteins with NEAr Transporter (NEAT) domains play a central role in heme acquisition and trafficking across the cell envelope of Gram-positive bacteria. Group A Streptococcus (GAS), a β-hemolytic human pathogen, expresses a NEAT protein, Shr, which binds several hemoproteins and extracellular matrix (ECM) components. Shr is a complex, membrane-anchored protein, with a unique N-terminal domain (NTD) and two NEAT domains separated by a central leucine-rich repeat region. In this study we have carried out an analysis of the functional domains in Shr. We show that Shr obtains heme in solution and furthermore reduces the heme iron; this is the first report of heme reduction by a NEAT protein. More specifically, we demonstrate that both of the constituent NEAT domains of Shr are responsible for binding heme, although they are missing a critical tyrosine residue found in the ligand-binding pocket of other heme-binding NEAT domains. Further investigations show that a previously undescribed region within the Shr NTD interacts with methemoglobin. Shr NEAT domains, however, do not contribute significantly to the binding of methemoglobin but mediate binding to the ECM components fibronectin and laminin. A protein fragment containing the NTD plus the first NEAT domain was found to be sufficient to sequester heme directly from methemoglobin. Correlating these in vitro findings to in vivo biological function, mutants analysis establishes the role of Shr in GAS growth with methemoglobin as a sole source of iron, and indicates that at least one NEAT domain is necessary for the utilization of methemoglobin. We suggest that Shr is the prototype of a new group of NEAT composite proteins involved in heme uptake found in pyogenic streptococci and Clostridium novyi. PMID:20807204

  18. Heme metabolism as an integral part of iron homeostasis

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    Paweł Lipiński

    2014-01-01

    Full Text Available Heme, a ferrous iron protoporphyrin IX complex, is employed as a prosthetic group in a number of diverse heme proteins that participate in important cellular and systemic physiological processes. Provision of an adequate amount of iron for heme biosynthesis is one of the elemental hallmarks of intracellular iron homeostasis. In the cell the bioavailability of iron for the two main iron biological pathways – heme synthesis and the biogenesis of iron-sulfur clusters ([Fe-S] – is mainly regulated by the IRP/IRE posttranscriptional system. The biogenesis of [Fe-S] centers is crucial for heme synthesis because these co-factors determine the activity of IRP1 and that of ferrochelatase, an enzyme responsible for the insertion of an iron into protoporphyrin IX to produce heme. On the other hand, delivery of iron for heme and hemoglobin synthesis in erythroblasts, precursors of erythrocytes in bone marrow, is an indispensable element of body iron homeostasis. This process relies on the recovery of iron from senescent red blood cells through the enzymatic degradation of heme molecules and recycling of iron to the circulation. Molecular coordination of these processes involves the activity of heme oxygenase 1, IRP1 and IRP2 as well as the functioning of the hepcidin-ferroportin regulatory axis. Recent studies show in mammals the existence of an expanded system of proteins involved in the transport of intact heme molecules at the cellular and systemic levels. The biological role of this system is of particular importance when the concentration of free heme reaches a toxic level in the body (intravascular hemolysis as well as locally in cells having intensive heme metabolism such as erythroblasts and macrophages.

  19. Heme A synthase in bacteria depends on one pair of cysteinyls for activity.

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    Lewin, Anna; Hederstedt, Lars

    2016-02-01

    Heme A is a prosthetic group unique for cytochrome a-type respiratory oxidases in mammals, plants and many microorganisms. The poorly understood integral membrane protein heme A synthase catalyzes the synthesis of heme A from heme O. In bacteria, but not in mitochondria, this enzyme contains one or two pairs of cysteine residues that are present in predicted hydrophilic polypeptide loops on the extracytoplasmic side of the membrane. We used heme A synthase from the eubacterium Bacillus subtilis and the hyperthermophilic archeon Aeropyrum pernix to investigate the functional role of these cysteine residues. Results with B. subtilis amino acid substituted proteins indicated the pair of cysteine residues in the loop connecting transmembrane segments I and II as being essential for catalysis but not required for binding of the enzyme substrate, heme O. Experiments with isolated A. pernix and B. subtilis heme A synthase demonstrated that a disulfide bond can form between the cysteine residues in the same loop and also between loops showing close proximity of the two loops in the folded enzyme protein. Based on the findings, we propose a classification scheme for the four discrete types of heme A synthase found so far in different organisms and propose that essential cysteinyls mediate transfer of reducing equivalents required for the oxygen-dependent catalysis of heme A synthesis from heme O. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Identification of a copper-repressible C-type heme protein of Methylococcus capsulatus (Bath). A member of a novel group of the bacterial di-heme cytochrome c peroxidase family of proteins.

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    Karlsen, Odd A; Kindingstad, Louise; Angelskår, Solveig M; Bruseth, Live J; Straume, Daniel; Puntervoll, Pål; Fjellbirkeland, Anne; Lillehaug, Johan R; Jensen, Harald B

    2005-12-01

    Genomic sequencing of the methanotrophic bacterium, Methylococcus capsulatus (Bath), revealed an open reading frame (MCA2590) immediately upstream of the previously described mopE gene (MCA2589). Sequence analyses of the deduced amino acid sequence demonstrated that the MCA2590-encoded protein shared significant, but restricted, sequence similarity to the bacterial di-heme cytochrome c peroxidase (BCCP) family of proteins. Two putative C-type heme-binding motifs were predicted, and confirmed by positive heme staining. Immunospecific recognition and biotinylation of whole cells combined with MS analyses confirmed expression of MCA2590 in M. capsulatus as a protein noncovalently associated with the cellular surface of the bacterium exposed to the cell exterior. Similar to MopE, expression of MCA2590 is regulated by the bioavailability of copper and is most abundant in M. capsulatus cultures grown under low copper conditions, thus indicating an important physiological role under these growth conditions. MCA2590 is distinguished from previously characterized members of the BCCP family by containing a much longer primary sequence that generates an increased distance between the two heme-binding motifs in its primary sequence. Furthermore, the surface localization of MCA2590 is in contrast to the periplasmic location of the reported BCCP members. Based on our experimental and bioinformatical analyses, we suggest that MCA2590 is a member of a novel group of bacterial di-heme cytochrome c peroxidases not previously characterized.

  1. Spectroscopic Studies Reveal That the Heme Regulatory Motifs of Heme Oxygenase-2 Are Dynamically Disordered and Exhibit Redox-Dependent Interaction with Heme

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    2015-01-01

    Heme oxygenase (HO) catalyzes a key step in heme homeostasis: the O2- and NADPH-cytochrome P450 reductase-dependent conversion of heme to biliverdin, Fe, and CO through a process in which the heme participates both as a prosthetic group and as a substrate. Mammals contain two isoforms of this enzyme, HO2 and HO1, which share the same α-helical fold forming the catalytic core and heme binding site, as well as a membrane spanning helix at their C-termini. However, unlike HO1, HO2 has an additional 30-residue N-terminus as well as two cysteine-proline sequences near the C-terminus that reside in heme regulatory motifs (HRMs). While the role of the additional N-terminal residues of HO2 is not yet understood, the HRMs have been proposed to reversibly form a thiol/disulfide redox switch that modulates the affinity of HO2 for ferric heme as a function of cellular redox poise. To further define the roles of the N- and C-terminal regions unique to HO2, we used multiple spectroscopic techniques to characterize these regions of the human HO2. Nuclear magnetic resonance spectroscopic experiments with HO2 demonstrate that, when the HRMs are in the oxidized state (HO2O), both the extra N-terminal and the C-terminal HRM-containing regions are disordered. However, protein NMR experiments illustrate that, under reducing conditions, the C-terminal region gains some structure as the Cys residues in the HRMs undergo reduction (HO2R) and, in experiments employing a diamagnetic protoporphyrin, suggest a redox-dependent interaction between the core and the HRM domains. Further, electron nuclear double resonance and X-ray absorption spectroscopic studies demonstrate that, upon reduction of the HRMs to the sulfhydryl form, a cysteine residue from the HRM region ligates to a ferric heme. Taken together with EPR measurements, which show the appearance of a new low-spin heme signal in reduced HO2, it appears that a cysteine residue(s) in the HRMs directly interacts with a second bound heme

  2. Radiosynthesis and biodistribution of a prosthetic group (¹⁸F-FENMA) conjugated to cyclic RGD peptides.

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    Olberg, Dag Erlend; Cuthbertson, Alan; Solbakken, Magne; Arukwe, Joseph M; Qu, Hong; Kristian, Alexandr; Bruheim, Skjalg; Hjelstuen, Ole Kristian

    2010-12-15

    We have recently reported a new N-methylaminooxy-based prosthetic group for the site-selective introduction of ¹⁸F-fluorine under mild acidic aqueous conditions into model peptides functionalized with a Michael acceptor moiety. To further investigate the utility of this methodology, the radiosynthesis of two cyclic RGD peptides was carried out, and in vivo biodistribution and microPET studies were performed in tumor-bearing mice. A cyclic RGD peptide was functionalized with the Michael acceptors trans-β-nitrostyrene carboxylic acid and 3-vinylsulfonylpropionic acid. Radiolabeling was then performed with the prosthetic group O-(2-(2-[¹⁸F]fluoroethoxy)ethyl)-N-methylhydroxylamine (¹⁸F-FENMA) yielding the ¹⁸F-conjugates in moderate yields (8.5-12%). Biodistribution, blocking, and microPET imaging studies were performed in a mouse xenograft model. The vinylsulfonyl-modified conjugate demonstrated good in vitro plasma stability. Biodistribution and microPET studies revealed excellent tumor uptake with low background in key organs and renal elimination as the predominant route of excretion. Blocking studies with coinjected nonlabeled RGD peptide confirmed the in vivo specificity for the integrin α(v)β₃. On the other hand, ¹⁸F-FENMA-nitrostyrene-RGD, although stable at conjugation pH 5, was found to rapidly degrade at physiological pH through loss of the ¹⁸F-prosthetic group.

  3. Heme on innate immunity and inflammation

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    Fabianno Ferreira Dutra

    2014-05-01

    Full Text Available Heme is an essential molecule expressed ubiquitously all through our tissues. Heme plays major functions in cellular physiology and metabolism as the prostetic group of diverse proteins. Once released from cells and from hemeproteins free heme causes oxidative damage and inflammation, thus acting as a prototypic damage-associated molecular pattern. In this context, free heme is a critical component of the pathological process of sterile and infectious hemolytic conditions including malaria, hemolytic anemias, ischemia-reperfusion and hemorrhage. The plasma scavanger proteins hemopexin and albumin reduce heme toxicity and are responsible for transporting free heme to intracellular compartments where it is catabolized by heme-oxygenase enzymes. Upon hemolysis or severe cellular damage the serum capacity to scavange heme may saturate and increase free heme to sufficient amounts to cause tissue damage in various organs. The mechanism by which heme causes reactive oxygen generation, activation of cells of the innate immune system and cell death are not fully understood. Although heme can directly promote lipid peroxidation by its iron atom, heme can also induce ROS generation and production of inflammatory mediators through the activation of selective signaling pathways. Heme activates innate immune cells such as macrophages and neutrophils through activation of innate immune receptors. The importance of these events has been demonstrated in infectious and non-infectious diseases models. In this review we will discuss the mechanisms behind heme-induced citotoxicity and inflammation and the consequences of these events on different tissues and diseases.

  4. Hemoglobin and heme scavenger receptors

    DEFF Research Database (Denmark)

    Nielsen, Marianne Jensby; Møller, Holger Jon; Moestrup, Søren Kragh

    2010-01-01

    Heme, the functional group of hemoglobin, myoglobin, and other hemoproteins, is a highly toxic substance when it appears in the extracellular milieu. To circumvent potential harmful effects of heme from hemoproteins released during physiological or pathological cell damage (such as hemolysis...

  5. Rehabilitation and Prosthetic Services

    Science.gov (United States)

    ... Sensory Aids Service » Prosthetic & Sensory Aids Service (PSAS) Rehabilitation and Prosthetic Services Menu Menu Rehabilitation and Prosthetics Rehabilitation and Prosthetic Services Home Amputation ...

  6. Heme Oxygenase-1 and breast cancer resistance protein protect against heme-induced toxicity

    NARCIS (Netherlands)

    Wagener, Frank A D T G; Dankers, Anita C A; van Summeren, Frank; Scharstuhl, Alwin; van den Heuvel, Jeroen J M W; Koenderink, Jan B; Pennings, Sebastiaan W C; Russel, Frans G M; Masereeuw, R.

    2013-01-01

    Heme is the functional group of diverse hemoproteins and crucial for many cellular processes. However, heme is increasingly recognized as a culprit for a wide variety of pathologies, including sepsis, malaria, and kidney failure. Excess of free heme can be detrimental to tissues by mediating

  7. Heme Oxygenase-1 and Breast Cancer Resistance Protein Protect Against Heme-induced Toxicity

    NARCIS (Netherlands)

    Wagener, F.A.D.T.G.; Dankers, A.C.A.; Summeren, F. van; Scharstuhl, A.; Heuvel, J.J. van den; Koenderink, J.B.; Pennings, S.W.C.; Russel, F.G.M.; Masereeuw, R.

    2013-01-01

    Heme is the functional group of diverse hemoproteins and crucial for many cellular processes. However, heme is increasingly recognized as a culprit for a wide variety of pathologies, including sepsis, malaria, and kidney failure. Excess of free heme can be detrimental to tissues by mediating

  8. SCMHBP: prediction and analysis of heme binding proteins using propensity scores of dipeptides.

    Science.gov (United States)

    Liou, Yi-Fan; Charoenkwan, Phasit; Srinivasulu, Yerukala; Vasylenko, Tamara; Lai, Shih-Chung; Lee, Hua-Chin; Chen, Yi-Hsiung; Huang, Hui-Ling; Ho, Shinn-Ying

    2014-01-01

    Heme binding proteins (HBPs) are metalloproteins that contain a heme ligand (an iron-porphyrin complex) as the prosthetic group. Several computational methods have been proposed to predict heme binding residues and thereby to understand the interactions between heme and its host proteins. However, few in silico methods for identifying HBPs have been proposed. This work proposes a scoring card method (SCM) based method (named SCMHBP) for predicting and analyzing HBPs from sequences. A balanced dataset of 747 HBPs (selected using a Gene Ontology term GO:0020037) and 747 non-HBPs (selected from 91,414 putative non-HBPs) with an identity of 25% was firstly established. Consequently, a set of scores that quantified the propensity of amino acids and dipeptides to be HBPs is estimated using SCM to maximize the predictive accuracy of SCMHBP. Finally, the informative physicochemical properties of 20 amino acids are identified by utilizing the estimated propensity scores to be used to categorize HBPs. The training and mean test accuracies of SCMHBP applied to three independent test datasets are 85.90% and 71.57%, respectively. SCMHBP performs well relative to comparison with such methods as support vector machine (SVM), decision tree J48, and Bayes classifiers. The putative non-HBPs with high sequence propensity scores are potential HBPs, which can be further validated by experimental confirmation. The propensity scores of individual amino acids and dipeptides are examined to elucidate the interactions between heme and its host proteins. The following characteristics of HBPs are derived from the propensity scores: 1) aromatic side chains are important to the effectiveness of specific HBP functions; 2) a hydrophobic environment is important in the interaction between heme and binding sites; and 3) the whole HBP has low flexibility whereas the heme binding residues are relatively flexible. SCMHBP yields knowledge that improves our understanding of HBPs rather than merely

  9. An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group.

    Science.gov (United States)

    Jeon, Jongho; Shim, Ha Eun; Mushtaq, Sajid; Choi, Mi Hee; Park, Sang Hyun; Choi, Dae Seong; Jang, Beom-Su

    2016-10-10

    Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient radiolabeling of DBCO-group-functionalized gold nanoparticles using a copper-free click reaction. Radioiodination of the stannylated precursor (2) was carried out by using [125I]NaI and chloramine T as an oxidant at room temperature for 15 min. After HPLC purification of the crude product, the purified 125I-labeled azide (1) was obtained with high radiochemical yield (75 ± 10%, n = 8) and excellent radiochemical purity (>99%). For the synthesis of radiolabeled 13-nm-sized gold nanoparticles, the DBCO-functionalized gold nanoparticles (3) were prepared by using a thiolated polyethylene glycol polymer. A copper-free click reaction between 1 and 3 gave the 125I-labeled gold nanoparticles (4) with more than 95% of radiochemical yield as determined by radio-thin-layer chromatography (radio-TLC). These results clearly indicate that the present radiolabeling method using a strain-promoted copper-free click reaction will be useful for the efficient and convenient radiolabeling of DBCO-group-containing nanomaterials.

  10. Prebiotics increase heme iron bioavailability and do not affect non-heme iron bioavailability in humans.

    Science.gov (United States)

    Weinborn, Valerie; Valenzuela, Carolina; Olivares, Manuel; Arredondo, Miguel; Weill, Ricardo; Pizarro, Fernando

    2017-05-24

    The aim of this study was to establish the effect of a prebiotic mix on heme and non-heme iron (Fe) bioavailability in humans. To this purpose, twenty-four healthy women were randomized into one of two study groups. One group ate one yogurt per day for 12 days with a prebiotic mix (prebiotic group) and the other group received the same yogurt but without the prebiotic mix (control group). Before and after the intake period, the subjects participated in Fe absorption studies. These studies used (55)Fe and (59)Fe radioactive isotopes as markers of heme Fe and non-heme Fe, respectively, and Fe absorption was measured by the incorporation of radioactive Fe into erythrocytes. The results showed that there were no significant differences in heme and non-heme Fe bioavailability in the control group. Heme Fe bioavailability of the prebiotic group increased significantly by 56% post-prebiotic intake. There were no significant differences in non-heme Fe bioavailability in this group. We concluded that daily consumption of a prebiotic mix increases heme Fe bioavailability and does not affect non-heme iron bioavailability.

  11. Separation and structure of the prosthetic group of the blue fluorescence protein from the bioluminescent bacterium Photobacterium phosphoreum

    Science.gov (United States)

    Koka, Prasad; Lee, John

    1979-01-01

    The highly fluorescent prosthetic group of the blue fluorescence protein purified from the bioluminescent bacterium Photobacterium phosphoreum has been dissociated and separated from its apoprotein by affinity chromatography on Cibacron Blue-Sepharose. It has been identified as 6,7-dimethyl-8-(1′-D-ribityl)lumazine by several methods of characterization, all of which gave results identical to those for an authentic sample. In neutral solution, absorption maxima are at 407, 275 (shoulder), and 256 nm, with a single fluorescence maximum at 491 nm. The proton magnetic resonance spectrum exhibits a singlet at 2.66 ppm corresponding to the methyl substituted at the 6 position of lumazine and a multiplet centered at 3.85 ppm corresponding to the C-2′-5′ protons of the ribityl group. A Raman spectrum was obtained by the technique of coherent anti-Stokes Raman scattering and the RF values by paper chromatography were determined in four solvent systems. The isolated compound was readily transformed into riboflavin by riboflavin synthetase. Fifty grams (dry weight) of P. phosphoreum contains at least 20 mg of this lumazine derivative, an amount comparable to that found in other microorganisms classified as riboflavin overproducers. The overproduction of this lumazine in this case apparently has to do with its function in the generation of bioluminescence. PMID:16592674

  12. Biglutaminyl-biliverdin IX alpha as a heme degradation product in the dengue fever insect-vector Aedes aegypti.

    Science.gov (United States)

    Pereira, Luiza O R; Oliveira, Pedro L; Almeida, Igor C; Paiva-Silva, Gabriela O

    2007-06-12

    Hemoglobin digestion in the midgut of hematophagous animals results in the release of its prosthetic group, heme, which is a pro-oxidant molecule. Heme enzymatic degradation is a protective mechanism that has been described in several organisms, including plants, bacteria, and mammals. This reaction is catalyzed by heme oxygenase and results in formation of carbon monoxide, ferrous ion, and biliverdin IXalpha. During digestion, a large amount of a green pigment is produced and secreted into the intestinal lumen of Aedes aegypti adult females. In the case of another blood-sucking insect, the kissing-bug Rhodnius prolixus, we have recently shown that heme degradation involves a complex pathway that generates dicysteinyl-biliverdin IX gamma. The light absorption spectrum of the Aedes purified pigment was similar to that of biliverdin, but its mobility on a reverse-phase chromatography column suggested a compound less hydrophobic than biliverdin IXalpha. Structural characterization by ESI-MS revealed that the mosquito pigment is the alpha isomer of biliverdin bound to two glutamine residues by an amide bond. This biglutaminyl-biliverdin is formed by oxidative cleavage of the heme porphyrin ring followed by two subsequent additions of glutamine residues to the biliverdin IXalpha. The role of this pathway in the adaptation of this insect vector to a blood-feeding habit is discussed.

  13. Density functional theory-broken symmetry (DFT-BS) methodology applied to electronic and magnetic properties of bioinorganic prosthetic groups.

    Science.gov (United States)

    Mouesca, Jean-Marie

    2014-01-01

    leading practitioner (for those interested, I invite them to look for his papers online (via Web of Knowledge for example) as they are simply too numerous to be cited here) of the BS methodology described below in the rich field of (bio)inorganic (transition metal) complexes and prosthetic groups and also a mentor (since my 2-year postdoctoral fellowship with him, back in 1992-1993) and a friend.

  14. Radiolabeling of phosphatidylserine-binding peptides with prosthetic groups N-[6-(4-[{sup 18}F]fluorobenzylidene)aminooxyhexyl]maleimide ([{sup 18}F]FBAM) and N-succinimidyl-4-[{sup 18}F]fluorobenzoate ([{sup 18}F]SFB)

    Energy Technology Data Exchange (ETDEWEB)

    Kapty, Janice, E-mail: jkapty@ualberta.ca [Department of Oncology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, T6G 1Z2 (Canada); Kniess, Torsten, E-mail: t.kniess@hzdr.de [Helmholtz-Zentrum Dresden-Rossendorf, Institute of Radiopharmacy, PF 510 119, 01314 Dresden (Germany); Wuest, Frank, E-mail: wuest@ualberta.ca [Department of Oncology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, T6G 1Z2 (Canada); Mercer, John R., E-mail: john.mercer@ualberta.ca [Department of Oncology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, T6G 1Z2 (Canada)

    2011-09-15

    The widely used {sup 18}F-prosthetic group N-succinimidyl-4-[{sup 18}F]fluorobenzoate ([{sup 18}F]SFB) and the recently developed N-[6-(4-[{sup 18}F]fluorobenzylidene)aminooxyhexyl]maleimide ([{sup 18}F]FBAM) were investigated for radiolabeling of two representative phosphatidylserine-binding peptides. The prosthetic groups were compared with respect to required reactions conditions for optimum labeling, radiolabeling yield and chemoselectivity. The N-terminus labeled product produced by reaction of [{sup 18}F]SFB with binding peptide LIKKPF was produced in 18% radiochemical yield while no N-terminus labeled product could be isolated following [{sup 18}F]SFB reaction with PDGLSR. When the peptides were modified by addition of a cysteine residue at the N-terminus they provided almost quantitative radiochemical yields with [{sup 18}F]FBAM. Results indicate that for the peptides in this study, [{sup 18}F]FBAM is a more useful prosthetic group compared to [{sup 18}F]SFB due to its excellent chemoselectivity and high radiochemical yield. - Highlights: > Phosphatidylserine-binding peptides were labeled with {sup 18}F. > The prosthetic groups N-[6-(4-[{sup 18}F]fluorobenzylidene)aminooxyhexyl]maleimide ([{sup 18}F]FBAM) and N-succinimidyl-4-[{sup 18}F]fluorobenzoate ([{sup 18}F]SFB) were compared for labeling selected peptides. > Purification of prosthetic groups by cartridge methods and by HPLC was compared.

  15. Porphyrin and heme metabolism and the porphyrias.

    Science.gov (United States)

    Bonkovsky, Herbert L; Guo, Jun-Tao; Hou, Weihong; Li, Ting; Narang, Tarun; Thapar, Manish

    2013-01-01

    Porphyrins and metalloporphyrins are the key pigments of life on earth as we know it, because they include chlorophyll (a magnesium-containing metalloporphyrin) and heme (iron protoporphyrin). In eukaryotes, porphyrins and heme are synthesized by a multistep pathway that involves eight enzymes. The first and rate-controlling step is the formation of delta-aminolevulinic acid (ALA) from glycine plus succinyl CoA, catalyzed by ALA synthase. Intermediate steps occur in the cytoplasm, with formation of the monopyrrole porphobilinogen and the tetrapyrroles hydroxymethylbilane and a series of porphyrinogens, which are serially decarboxylated. Heme is utilized chiefly for the formation of hemoglobin in erythrocytes, myoglobin in muscle cells, cytochromes P-450 and mitochondrial cytochromes, and other hemoproteins in hepatocytes. The rate-controlling step of heme breakdown is catalyzed by heme oxygenase (HMOX), of which there are two isoforms, called HMOX1 and HMOX2. HMOX breaks down heme to form biliverdin, carbon monoxide, and iron. The porphyrias are a group of disorders, mainly inherited, in which there are defects in normal porphyrin and heme synthesis. The cardinal clinical features are cutaneous (due to the skin-damaging effects of excess deposited porphyrins) or neurovisceral attacks of pain, sometimes with weakness, delirium, seizures, and the like (probably due mainly to neurotoxic effects of ALA). The treatment of choice for the acute hepatic porphyrias is intravenous heme therapy, which repletes a critical regulatory heme pool in hepatocytes and leads to downregulation of hepatic ALA synthase, which is a biochemical hallmark of all forms of acute porphyria in relapse.

  16. Prosthetic stomatitis with removable dentures

    Directory of Open Access Journals (Sweden)

    Rozalieva Yu.Yu.

    2012-06-01

    Full Text Available The Research Objective: To study patients with prosthetic stomatitis, who use the removable laminar dentures. Materials: The consultations and treatment of 79 patients aged 47-65 years have been conducted. The patients have been divided into two clinical groups. The first clinical group (39 persons with the performance of immediate prosthet-ics; the second control clinical group (40 persons — the permanent dentures were produced without the preliminary instruction. Results: All the patients, having the laminar dentures without the preliminary use of immediate constructions of dentures, in spite of repeated correction of them, have had changes of dentures and transitory fold. Patients have been exposed to prosthetic stomatitis of different etiology (without trauma; the single-shot or multiple correction of dentures by the method of rebasing with using of cold cure plastics has been made. Conclusion: Structural and functional changes of dentition during the prosthetic stomatitis lead to disorders, associated by the mucositis. Use of the term of «prosthetic stomatitis» reflects etiological and pathogenetic component of changes in the denture-supporting tissues

  17. Molecular biological study of early steps of heme biosynthesis in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Li, Jianming.

    1989-01-01

    The heme molecule is the prosthetic group of hemoglobins, cytochromes, catalases and peroxidases, and modified tetrapyrroles are the active moieties of the chlorophylls and vitamin B{sub 12}. The pathway of heme biosynthesis is highly conserved except for the formation of 5-aminolevulinic acid (ALA), the first common intermediate. ALA can be synthesized from glutamic acid (C{sub 5} pathway) or succinyl CoA and glycine (C{sub 4} pathway). In E. coli, the heme biosynthetic pathway consists of more than eight enzymatic steps. The genes encoding these enzymes are widely scattered on the chromosome. An E. coli heme-requiring, hemin-permeable mutant had no detectable 5-aminolevulinic acid dehydratase (ALA D) or porphobilinogen deaminase (PBG D) activities. The gene which complemented this mutation was cloned. PBG D activity was restored to normal levels, but the activity of ALA D was 20-30 fold higher than normal. A maxicell procedure confirmed that the cloned gene was hemB. The hemB gene was sequenced. Two promoter regions, two Shine-Dalgarno sequences and two possible initiation sites were identified. Extensive homologies with yeast (36%), human liver (40%) and rat liver (40%) amino acid sequences were observed, especially in the sixteen-amino acid Zn-binding region (75%) and the four amino acids surrounding the essential lysine at the active site (100% for rate and human proteins). Analysis of promoter strength and two independent analyses of codon usage indicated that the hemB gene is moderately-expressed. E. coli hemA gene was cloned and sequenced. Complemented mutants overproduced 5-ALA and porphyrins. The cloned sequence appears to encode a 46 kDa protein. The amino acid sequence of the cloned gene product showed no significant homologies with any cloned ALA synthase nor with any protein in two data banks.

  18. Supramolecular assembling systems formed by heme-heme pocket interactions in hemoproteins.

    Science.gov (United States)

    Oohora, Koji; Onoda, Akira; Hayashi, Takashi

    2012-12-14

    A native protein in a biological system spontaneously produces large and elegant assemblies via self-assembly or assembly with various biomolecules which provide non-covalent interactions. In this context, the protein plays a key role in construction of a unique supramolecular structure operating as a functional system. Our group has recently highlighted the structure and function of hemoproteins reconstituted with artificially created heme analogs. The heme molecule is a replaceable cofactor of several hemoproteins. Here, we focus on the successive supramolecular protein assemblies driven by heme-heme pocket interactions to afford various examples of protein fibers, networks and three-dimensional clusters in which an artificial heme moiety is introduced onto the surface of a hemoprotein via covalent linkage and the native heme cofactor is removed from the heme pocket. This strategy is found to be useful for constructing hybrid materials with an electrode or with nanoparticles. The new systems described herein are expected to lead to the generation of various biomaterials with functions and characteristic physicochemical properties similar to those of hemoproteins.

  19. Targeting Prostate-Specific Membrane Antigen (PSMA) with F-18-Labeled Compounds: the Influence of Prosthetic Groups on Tumor Uptake and Clearance Profile.

    Science.gov (United States)

    Bouvet, Vincent; Wuest, Melinda; Bailey, Justin J; Bergman, Cody; Janzen, Nancy; Valliant, John F; Wuest, Frank

    2017-12-01

    Prostate-specific membrane antigen (PSMA) is an important biomarker expressed in the majority of prostate cancers. The favorable positron emission tomography (PET) imaging profile of the PSMA imaging agent 2-(3-(1-carboxy-5-[(6-[(18)F]fluoro-pyridine-3-carbonyl)-amino]-pentyl)-ureido)-pentane-dioic acid [(18)F]DCFPyL in preclinical prostate cancer models and in prostate cancer patients stimulated the development and validation of other fluorine-containing PSMA inhibitors to further enhance pharmacokinetics and simplify production methods. Here, we describe the synthesis and radiopharmacological evaluation of various F-18-labeled PSMA inhibitors which were prepared through different prosthetic group chemistry strategies. Prosthetic groups N-succinimidyl-4-[(18)F]fluorobenzoate ([(18)F]SFB), 4-[(18)F]fluorobenzaldehyde, and 2-deoxy-2-[(18)F]fluoro-D-glucose ([(18)F]FDG) were used for bioconjugation reactions to PSMA-binding lysine-urea-glutamate scaffold via acylation and oxime formation. All fluorine-containing PSMA inhibitors were tested for their PSMA inhibitory potency in an in vitro competitive binding assay in comparison to an established reference compound [(125)I]TAAG-PSMA. Tumor uptake and clearance profiles of three F-18-labeled PSMA inhibitors ([(18)F]4, [(18)F]7, and [(18)F]8) were studied with dynamic PET imaging using LNCaP tumor-bearing mice. F-18-labeled PSMA inhibitors were synthesized in 32-69 % radiochemical yields using (1) acylation reaction at the primary amino group of the lysine residue with [(18)F]SFB and (2) oxime formation with 4-[(18)F]fluorobenzaldehyde and [(18)F]FDG using the respective aminooxy-functionalized lysine residue. Compound 7 displayed an IC50 value of 6 nM reflecting very high affinity for PSMA. Compounds 4 and 8 showed IC50 values of 13 and 62 nM, respectively. The IC50 value of reference compound DCFPyL was 13 nM. Dynamic PET imaging revealed the following SUV60min for radiotracer uptake in PSMA(+) LNCaP tumors: 0

  20. A simple and reusable fluorescent sensor for heme proteins based on a conjugated polymer-doped electrospun nanofibrous membrane.

    Science.gov (United States)

    Wang, Huaming; Peng, Zhou; Long, Yuanyuan; Chen, Haibo; Yang, Yufei; Li, Na; Liu, Feng

    2012-05-30

    We reported a simple and reusable fluorescent sensor for heme proteins based on the electrospun nanofibrous membrane doped with a fluorescent conjugated polymer P. The sensor showed favorable fluorescence sensing performance towards the heme proteins, including hemoglobin (Hb), myoglobin (Mb) and cytochrome c (Cyt c). The surface wettability and sensing performance of the electrospun nanofibrous membrane were investigated in detail using Hb as the model. The nanofibrous sensor showed satisfactory reversibility with less than 10% signal loss after nine quenching-regeneration cycles, and good batch-to-batch reproducibility with a relative standard deviation of 3.4% (n=3). The linear range of the sensor for Hb determination was 2.0×10(-8) to 3.0×10(-6) M with a detection limit of 1.2×10(-8) M. The quenching process is mainly based on the fluorescence resonance energy transfer mechanism between the fluorescent conjugated polymer P and the heme prosthetic groups, therefore the sensor was selective against most of the common interferents. As an example to evaluate the feasibility of the sensor in practical application, Hb in human blood samples was determined and the results were in good agreement with the data provided by the hospital. To the best of our knowledge, this is the first work using fluorescent electrospun nanofibrous sensor for protein analysis in real biological sample. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. AI and Prosthetics

    OpenAIRE

    Kyriazi, Nefeli Evdokia

    2016-01-01

    Prosthetics are very important to an amputee.The introduction of technology to prosthetics has allowed bionic limbs to emerge and change the way we were thinking about prosthetic limbs.More and more companies create new innovative models,but not affordable for anyone.3D Printing gives more options.

  2. Heme Recognition By a Staphylococcus Aureus IsdE

    Energy Technology Data Exchange (ETDEWEB)

    Grigg, J.C.; Vermeiren, C.L.; Heinrichs, D.E.; Murphy, M.E.P.

    2009-06-03

    Staphylococcus aureus is a Gram-positive bacterial pathogen and a leading cause of hospital acquired infections. Because the free iron concentration in the human body is too low to support growth, S. aureus must acquire iron from host sources. Heme iron is the most prevalent iron reservoir in the human body and a predominant source of iron for S. aureus. The iron-regulated surface determinant (Isd) system removes heme from host heme proteins and transfers it to IsdE, the cognate substrate-binding lipoprotein of an ATP-binding cassette transporter, for import and subsequent degradation. Herein, we report the crystal structure of the soluble portion of the IsdE lipoprotein in complex with heme. The structure reveals a bi-lobed topology formed by an N- and C-terminal domain bridged by a single {alpha}-helix. The structure places IsdE as a member of the helical backbone metal receptor superfamily. A six-coordinate heme molecule is bound in the groove established at the domain interface, and the heme iron is coordinated in a novel fashion for heme transporters by Met{sup 78} and His{sup 229}. Both heme propionate groups are secured by H-bonds to IsdE main chain and side chain groups. Of these residues, His{sup 299} is essential for IsdE-mediated heme uptake by S. aureus when growth on heme as a sole iron source is measured. Multiple sequence alignments of homologues from several other Gram-positive bacteria, including the human pathogens pyogenes, Bacillus anthracis, and Listeria monocytogenes, suggest that these other systems function equivalently to S. aureus IsdE with respect to heme binding and transport.

  3. Structure of the Escherichia coli O157:H7 heme oxygenase ChuS in complex with heme and enzymatic inactivation by mutation of the heme coordinating residue His-193

    Energy Technology Data Exchange (ETDEWEB)

    Suits,M.; Jaffer, N.; Jia, Z.

    2006-01-01

    Heme oxygenases catalyze the oxidation of heme to biliverdin, CO, and free iron. For pathogenic microorganisms, heme uptake and degradation are critical mechanisms for iron acquisition that enable multiplication and survival within hosts they invade. Here we report the first crystal structure of the pathogenic Escherichia coli O157:H7 heme oxygenase ChuS in complex with heme at 1.45 {angstrom} resolution. When compared with other heme oxygenases, ChuS has a unique fold, including structural repeats and a {beta}-sheet core. Not surprisingly, the mode of heme coordination by ChuS is also distinct, whereby heme is largely stabilized by residues from the C-terminal domain, assisted by a distant arginine from the N-terminal domain. Upon heme binding, there is no large conformational change beyond the fine tuning of a key histidine (His-193) residue. Most intriguingly, in contrast to other heme oxygenases, the propionic side chains of heme are orientated toward the protein core, exposing the {alpha}-meso carbon position where O{sub 2} is added during heme degradation. This unique orientation may facilitate presentation to an electron donor, explaining the significantly reduced concentration of ascorbic acid needed for the reaction. Based on the ChuS-heme structure, we converted the histidine residue responsible for axial coordination of the heme group to an asparagine residue (H193N), as well as converting a second histidine to an alanine residue (H73A) for comparison purposes. We employed spectral analysis and CO measurement by gas chromatography to analyze catalysis by ChuS, H193N, and H73A, demonstrating that His-193 is the key residue for the heme-degrading activity of ChuS.

  4. Covalent heme attachment to the protein in human heme oxygenase-1 with selenocysteine replacing the His25 proximal iron ligand.

    Science.gov (United States)

    Jiang, Yongying; Trnka, Michael J; Medzihradszky, Katalin F; Ouellet, Hugues; Wang, Yongqiang; Ortiz de Montellano, Paul R

    2009-03-01

    To characterize heme oxygenase with a selenocysteine (SeCys) as the proximal iron ligand, we have expressed truncated human heme oxygenase-1 (hHO-1) His25Cys, in which Cys-25 is the only cysteine, in the Escherichia coli cysteine auxotroph strain BL21(DE3)cys. Selenocysteine incorporation into the protein was demonstrated by both intact protein mass measurement and mass spectrometric identification of the selenocysteine-containing tryptic peptide. One selenocysteine was incorporated into approximately 95% of the expressed protein. Formation of an adduct with Ellman's reagent (DTNB) indicated that the selenocysteine in the expressed protein was in the reduced state. The heme-His25SeCys hHO-1 complex could be prepared by either (a) supplementing the overexpression medium with heme, or (b) reconstituting the purified apoprotein with heme. Under reducing conditions in the presence of imidazole, a covalent bond is formed by addition of the selenocysteine residue to one of the heme vinyl groups. No covalent bond is formed when the heme is replaced by mesoheme, in which the vinyls are replaced by ethyl groups. These results, together with our earlier demonstration that external selenolate ligands can transfer an electron to the iron [Y. Jiang, P.R. Ortiz de Montellano, Inorg. Chem. 47 (2008) 3480-3482 ], indicate that a selenyl radical is formed in the hHO-1 His25SeCys mutant that adds to a heme vinyl group.

  5. Immunogenicity of heme complexes of peptides designed to mimic the heme environment of myoglobin and hemoglobin.

    Science.gov (United States)

    Atassi, M Zouhair; Childress, Catherine

    2005-01-01

    In the preceding paper (Protein J. 25, pages 37-49, 2005), we reported the preparation and oxygen-binding properties of peptides that form stable complexes with heme mimic. The design of the peptides was based on the natural environment of the heme group in myoglobin (Mb) and in the alpha- and beta-subunits of human adult hemoglobin (Hb). In the present work, the heme-peptides were each administered into mice, either as emulsions in adjuvant (both for injections and boosters) or intravenously as solutions in phosphate-buffered saline. Antibody (Ab) responses, monitored up to 14 weeks after the first administration, showed that when the heme-peptides were injected with adjuvant they stimulated Ab responses against the immunizing peptide, which in most cases bound to the correlate protein (Mb or Hb). However these heme-peptides were non-immunogenic when administered in PBS intravenously. It is concluded that heme-peptides:(a) would not trigger an adverse immune response if used for transfusion purposes.

  6. Tyrosine oxidation in heme oxygenase: examination of long-range proton-coupled electron transfer.

    Science.gov (United States)

    Smirnov, Valeriy V; Roth, Justine P

    2014-10-01

    Heme oxygenase is responsible for the degradation of a histidine-ligated ferric protoporphyrin IX (Por) to biliverdin, CO, and the free ferrous ion. Described here are studies of tyrosyl radical formation reactions that occur after oxidizing Fe(III)(Por) to Fe(IV)=O(Por(·+)) in human heme oxygenase isoform-1 (hHO-1) and the structurally homologous protein from Corynebacterium diphtheriae (cdHO). Site-directed mutagenesis on hHO-1 probes the reduction of Fe(IV)=O(Por(·+)) by tyrosine residues within 11 Å of the prosthetic group. In hHO-1, Y58· is implicated as the most likely site of oxidation, based on the pH and pD dependent kinetics. The absence of solvent deuterium isotope effects in basic solutions of hHO-1 and cdHO contrasts with the behavior of these proteins in the acidic solution, suggesting that long-range proton-coupled electron transfer predominates over electron transfer.

  7. Magnetic resonance spectral characterization of the heme active site of Coprinus cinereus peroxidase

    Energy Technology Data Exchange (ETDEWEB)

    Lukat, G.S.; Rodgers, K.R.; Jabro, M.N.; Goff, H.M. (Univ. of Iowa, Iowa City (USA))

    1989-04-18

    Examination of the peroxidase isolated from the inkcap Basidiomycete Coprinus cinereus shows that the 42,000-dalton enzyme contains a protoheme IX prosthetic group. Reactivity assays and the electronic absorption spectra of native Coprinus peroxidase and several of its ligand complexes indicate that this enzyme has characteristics similar to those reported for horseradish peroxidase. In this paper, the authors characterize the H{sub 2}O{sub 2}-oxidized forms of Coprinus peroxidase compounds I, II, and III by electronic absorption and magnetic resonance spectroscopies. Electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) studies of this Coprinus peroxidase indicate the presence of high-spin Fe(III) in the native protein and a number of differences between the heme site of Coprinus peroxidase and horseradish peroxidase. Carbon-13 (of the ferrous CO adduct) and nitrogen-15 (of the cyanide complex) NMR studies together with proton NMR studies of the native and cyanide-complexed Caprinus peroxidase are consistent with coordination of a proximal histidine ligand. The EPR spectrum of the ferrous NO complex is also reported. Protein reconstitution with deuterated hemin has facilitated the assignment of the heme methyl resonances in the proton NMR spectrum.

  8. Influence of the covalent heme-protein bonds on the redox thermodynamics of human myeloperoxidase.

    Science.gov (United States)

    Battistuzzi, Gianantonio; Stampler, Johanna; Bellei, Marzia; Vlasits, Jutta; Soudi, Monika; Furtmüller, Paul G; Obinger, Christian

    2011-09-20

    Myeloperoxidase (MPO) is the most abundant neutrophil enzyme and catalyzes predominantly the two-electron oxidation of ubiquitous chloride to generate the potent bleaching hypochlorous acid, thus contributing to pathogen killing as well as inflammatory diseases. Its catalytic properties are closely related with unique posttranslational modifications of its prosthetic group. In MPO, modified heme b is covalently bound to the protein via two ester linkages and one sulfonium ion linkage with a strong impact on its (electronic) structure and biophysical and chemical properties. Here, the thermodynamics of the one-electron reduction of the ferric heme in wild-type recombinant MPO and variants with disrupted heme-protein bonds (M243V, E242Q, and D94V) have been investigated by thin-layer spectroelectrochemistry. It turns out that neither the oligomeric structure nor the N-terminal extension in recombinant MPO modifies the peculiar positive reduction potential (E°' = 0.001 V at 25 °C and pH 7.0) or the enthalpy or entropy of the Fe(III) to Fe(II) reduction. By contrast, upon disruption of the MPO-typical sulfonium ion linkage, the reduction potential is significantly lower (-0.182 V). The M243V mutant has an enthalpically stabilized ferric state, whereas its ferrous form is entropically favored because of the loss of rigidity of the distal H-bonding network. Exchange of an adjacent ester bond (E242Q) induced similar but less pronounced effects (E°' = -0.094 V), whereas in the D94V variant (E°' = -0.060 V), formation of the ferrous state is entropically disfavored. These findings are discussed with respect to the chlorination and bromination activity of the wild-type protein and the mutants. © 2011 American Chemical Society

  9. Threshold concepts in prosthetics

    OpenAIRE

    Hill, Sophie

    2016-01-01

    Background: Curriculum documents identify key concepts within learning prosthetics. Threshold concepts provide an alternative way of viewing the curriculum, focussing on the ways of thinking and practicing within prosthetics. Threshold concepts can be described as an opening to a different way of viewing a concept. This article forms part of a larger study exploring what students and staff experience as difficult in learning about prosthetics. Objectives: To explore possible thresh...

  10. Prevention of Prosthetic Dentistry

    Directory of Open Access Journals (Sweden)

    Eremin O.V.

    2011-03-01

    Full Text Available Prevention in prosthetic dentistry is not just a regular oral hygiene and the prevention of caries in the early stages of its development. The initial goal of orthopedic and dental should be the ability to convey to the patient's sense of pros-thetics that proteziruya one saved more. An example is included prosthetic dental arch defects with bridges or single artificial crowns on implants that will prevent movement of teeth and the continuity of the dentition

  11. How Heme Oxygenase-1 Prevents Heme-Induced Cell Death.

    Directory of Open Access Journals (Sweden)

    Lilibeth Lanceta

    Full Text Available Earlier observations indicate that free heme is selectively toxic to cells lacking heme oxygenase-1 (HO-1 but how this enzyme prevents heme toxicity remains unexplained. Here, using A549 (human lung cancer and immortalized human bronchial epithelial cells incubated with exogenous heme, we find knock-down of HO-1 using siRNA does promote the accumulation of cell-associated heme and heme-induced cell death. However, it appears that the toxic effects of heme are exerted by "loose" (probably intralysosomal iron because cytotoxic effects of heme are lessened by pre-incubation of HO-1 deficient cells with desferrioxamine (which localizes preferentially in the lysosomal compartment. Desferrioxamine also decreases lysosomal rupture promoted by intracellularly generated hydrogen peroxide. Supporting the importance of endogenous oxidant production, both chemical and siRNA inhibition of catalase activity predisposes HO-1 deficient cells to heme-mediated killing. Importantly, it appears that HO-1 deficiency somehow blocks the induction of ferritin; control cells exposed to heme show ~10-fold increases in ferritin heavy chain expression whereas in heme-exposed HO-1 deficient cells ferritin expression is unchanged. Finally, overexpression of ferritin H chain in HO-1 deficient cells completely prevents heme-induced cytotoxicity. Although two other products of HO-1 activity--CO and bilirubin--have been invoked to explain HO-1-mediated cytoprotection, we conclude that, at least in this experimental system, HO-1 activity triggers the induction of ferritin and the latter is actually responsible for the cytoprotective effects of HO-1 activity.

  12. Threshold concepts in prosthetics.

    Science.gov (United States)

    Hill, Sophie

    2017-12-01

    Curriculum documents identify key concepts within learning prosthetics. Threshold concepts provide an alternative way of viewing the curriculum, focussing on the ways of thinking and practicing within prosthetics. Threshold concepts can be described as an opening to a different way of viewing a concept. This article forms part of a larger study exploring what students and staff experience as difficult in learning about prosthetics. To explore possible threshold concepts within prosthetics. Qualitative, interpretative phenomenological analysis. Data from 18 students and 8 staff at two universities with undergraduate prosthetics and orthotics programmes were generated through interviews and questionnaires. The data were analysed using an interpretative phenomenological analysis approach. Three possible threshold concepts arose from the data: 'how we walk', 'learning to talk' and 'considering the person'. Three potential threshold concepts in prosthetics are suggested with possible implications for prosthetics education. These possible threshold concepts involve changes in both conceptual and ontological knowledge, integrating into the persona of the individual. This integration occurs through the development of memories associated with procedural concepts that combine with disciplinary concepts. Considering the prosthetics curriculum through the lens of threshold concepts enables a focus on how students learn to become prosthetists. Clinical relevance This study provides new insights into how prosthetists learn. This has implications for curriculum design in prosthetics education.

  13. Structure and Heme-Independent Peroxidase Activity of a Fully-Coordinated Mononuclear Mn(II) Complex with a Schiff-Base Tripodal Ligand Containing Three Imidazole Groups

    Energy Technology Data Exchange (ETDEWEB)

    Sarkar, Shuranjan; Lee, Hong In [Kyungpook National University, Daegu (Korea, Republic of); Moon, Do Hyun [Pohang Accelerator Laboratory, Pohang (Korea, Republic of); Lah, Myoung Soo [Ulsan National Institute of Science and Technology, Ulsan (Korea, Republic of)

    2010-11-15

    New complex [Mn(II)H{sub 1.5}L]{sub 2}[Mn(II)H{sub 3}L]{sub 2}(ClO{sub 4}){sub 5}·3H{sub 2}O, where H{sub 3}L is tris{2-(4-imidazolyl)methyliminoethyl} amine (imtren), has been prepared by reacting manganese(II) perchlorate hexahydrate with the imtren ligand in methanol. X-ray crystallographic study revealed that the imtren ligand hexadentately binds to Mn(II) ion through the three Schiff-base imine N atoms and three imidazole N atoms with a distorted octahedral geometry, and the apical tertiary amine N atom of the ligand pseudo-coordinates to Mn(II), forming overall a pseudo-seven coordination environment. The hydrogen-bonds between imidazole and imidazolate of [Mn(II)H{sub 1.5}L]{sup 0.5+} complex ions are extended to build a 2D puckered network with trigonal voids. [Mn(II)H{sub 3}L]{sup 2+} complex ions constitutes another extended 2D puckered layer without hydrogen bonds. Two layers are wedged each other to constitute overall stack of the crystal. Peroxidase activity of complex 1 was examined by observing the oxidation of 2,2'-azinobis(3-ethylbenzothiazoline)- 6-sulfonic acid (ABTS) with hydrogen peroxide in the presence of complex 1. Generation of ABTS{sup +·} was observed by UV-vis and EPR spectroscopies, indicating that the complex 1, a fully-coordinated mononuclear Mn(II) complex with nitrogen-only ligand, has a heme-independent peroxidase activity.

  14. Introduction of a covalent histidine-heme linkage in a hemoglobin: a promising tool for heme protein engineering.

    Science.gov (United States)

    Rice, Selena L; Preimesberger, Matthew R; Johnson, Eric A; Lecomte, Juliette T J

    2014-12-01

    The hemoglobins of the cyanobacteria Synechococcus and Synechocystis (GlbNs) are capable of spontaneous and irreversible attachment of the b heme to the protein matrix. The reaction, which saturates the heme 2-vinyl by addition of a histidine residue, is reproduced in vitro by preparing the recombinant apoprotein, adding ferric heme, and reducing the iron to the ferrous state. Spontaneous covalent attachment of the heme is potentially useful for protein engineering purposes. Thus, to explore whether the histidine-heme linkage can serve in such applications, we attempted to introduce it in a test protein. We selected as our target the heme domain of Chlamydomonas eugametos LI637 (CtrHb), a eukaryotic globin that exhibits less than 50% sequence identity with the cyanobacterial GlbNs. We chose two positions, 75 in the FG corner and 111 in the H helix, to situate a histidine near a vinyl group. We characterized the proteins with gel electrophoresis, absorbance spectroscopy, and NMR analysis. Both T111H and L75H CtrHbs reacted upon reduction of the ferric starting material containing cyanide as the distal ligand to the iron. With L75H CtrHb, nearly complete (>90%) crosslinking was observed to the 4-vinyl as expected from the X-ray structure of wild-type CtrHb. Reaction of T111H CtrHb also occurred at the 4-vinyl, in a 60% yield indicating a preference for the flipped heme orientation in the starting material. The work suggests that the His-heme modification will be applicable to the design of proteins with a non-dissociable heme group. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. The heme-regulatory motif of nuclear receptor Rev-erbβ is a key mediator of heme and redox signaling in circadian rhythm maintenance and metabolism.

    Science.gov (United States)

    Carter, Eric L; Ramirez, Yanil; Ragsdale, Stephen W

    2017-07-07

    Rev-erbβ is a heme-responsive transcription factor that regulates genes involved in circadian rhythm maintenance and metabolism, effectively bridging these critical cellular processes. Heme binding to Rev-erbβ indirectly facilitates its interaction with the nuclear receptor co-repressor (NCoR1), resulting in repression of Rev-erbβ target genes. Fe 3+ -heme binds in a 6-coordinate complex with axial His and Cys ligands, the latter provided by a heme-regulatory motif (HRM). Rev-erbβ was thought to be a heme sensor based on a weak K d value for the Rev-erbβ·heme complex of 2 μm determined with isothermal titration calorimetry. However, our group demonstrated with UV-visible difference titrations that the K d value is in the low nanomolar range, and the Fe 3+ -heme off-rate is on the order of 10 -6 s -1 making Rev-erbβ ineffective as a sensor of Fe 3+ -heme. In this study, we dissected the kinetics of heme binding to Rev-erbβ and provided a K d for Fe 3+ -heme of ∼0.1 nm Loss of the HRM axial thiolate via redox processes, including oxidation to a disulfide with a neighboring cysteine or dissociation upon reduction of Fe 3+ - to Fe 2+ -heme, decreased binding affinity by >20-fold. Furthermore, as measured in a co-immunoprecipitation assay, substitution of the His or Cys heme ligands in Rev-erbβ was accompanied by a significant loss of NCoR1 binding. These results demonstrate the importance of the Rev-erbβ HRM in regulating interactions with heme and NCoR1 and advance our understanding of how signaling through HRMs affects the major cellular processes of circadian rhythm maintenance and metabolism. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  16. Hepatic heme catabolism in cultured hepatocytes

    Energy Technology Data Exchange (ETDEWEB)

    Lincoln, B.C.; Bonkovsky, H.L.

    1987-05-01

    Uncertainty persists concerning the role and importance of heme oxygenase in the catabolism of heme by hepatocytes. The products of heme oxygenase catalyzed heme catabolism are equimolar amounts of biliverdin IX..cap alpha.., CO, and iron. Previous reports from studies with rodent hepatocyte cultures have suggested the possibility that non-heme oxygenase pathway(s) predominate in the breakdown of hepatic hemoprotein heme. The authors have studied this question in cultured chick embryo hepatocytes, which retain normal regulation of heme metabolism and levels of cytochromes P-450 as in intact animals. Exogenous heme added to the culture medium with control chick embryo hepatocyte cultures was quantitatively converted to biliverdin IX..cap alpha... To study endogenous heme breakdown, cellular heme was labelled by exposing cultured cells to (5-/sup 14/C) 5-aminolevulinic acid (ALA). The hepatocytes were also treated with mephenytoin that increases cytochrome P-450, total hepatic heme and heme oxygenase. At various times after labelling heme, biliverdin, and CO were isolated and counted. For at least 8 hrs, the increase in CO radioactivity corresponded to the loss of radioactivity in heme. Beyond 1 h biliverdin was unstable in culture medium, but for 1 h after labelling (dpm BVIX..cap alpha.. + dpm CO) ..delta..dpm heme. All BV detected was the ..cap alpha.. isomer. They conclude that heme oxygenase accounts for both endogenous and exogenous heme breakdown by hepatocytes.

  17. GIANT PROSTHETIC VALVE THROMBUS

    Directory of Open Access Journals (Sweden)

    Prashanth Kumar

    2015-04-01

    Full Text Available Mechanical prosthetic valves are predisposed to bleeding, thrombosis & thromboembolic complications. Overall incidence of thromboembolic complications is 1% per year who are on oral anticoagulants, whereas bleeding complications incidence is 0.5% to 6.6% per year. 1, 2 Minimization of Scylla of thromboembolic & Charybdis of bleeding complication needs a balancing act of optimal antithrombotic therapy. We are reporting a case of middle aged male patient with prosthetic mitral valve presenting in heart failure. Patient had discontinued anticoagulants, as he had subdural hematoma in the past. He presented to our institute with a giant prosthetic valve thrombus.

  18. The 1.5-Å Structure of XplA-heme, an Unusual Cytochrome P450 Heme Domain That Catalyzes Reductive Biotransformation of Royal Demolition Explosive*

    Science.gov (United States)

    Sabbadin, Federico; Jackson, Rosamond; Haider, Kamran; Tampi, Girish; Turkenburg, Johan P.; Hart, Sam; Bruce, Neil C.; Grogan, Gideon

    2009-01-01

    XplA is a cytochrome P450 of unique structural organization, consisting of a heme- domain that is C-terminally fused to its native flavodoxin redox partner. XplA, along with flavodoxin reductase XplB, has been shown to catalyze the breakdown of the nitramine explosive and pollutant hexahydro-1,3,5-trinitro-1,3,5-triazine (royal demolition explosive) by reductive denitration. The structure of the heme domain of XplA (XplA-heme) has been solved in two crystal forms: as a dimer in space group P21 to a resolution of 1.9 Å and as a monomer in space group P21212 to a resolution of 1.5 Å, with the ligand imidazole bound at the heme iron. Although it shares the overall fold of cytochromes P450 of known structure, XplA-heme is unusual in that the kinked I-helix that traverses the distal face of the heme is broken by Met-394 and Ala-395 in place of the well conserved Asp/Glu plus Thr/Ser, important in oxidative P450s for the scission of the dioxygen bond prior to substrate oxygenation. The heme environment of XplA-heme is hydrophobic, featuring a cluster of three methionines above the heme, including Met-394. Imidazole was observed bound to the heme iron and is in close proximity to the side chain of Gln-438, which is situated over the distal face of the heme. Imidazole is also hydrogen-bonded to a water molecule that sits in place of the threonine side-chain hydroxyl exemplified by Thr-252 in Cyt-P450cam. Both Gln-438 → Ala and Ala-395 → Thr mutants of XplA-heme displayed markedly reduced activity compared with the wild type for royal demolition explosive degradation when combined with surrogate electron donors. PMID:19692330

  19. Alteration of the Regiospecificity of Human Heme Oxygenase-1 by Unseating of the Heme but not Disruption of the Distal Hydrogen Bonding Network†

    Science.gov (United States)

    Wang, Jinling; Evans, John P.; Ogura, Hiroshi; La Mar, Gerd N.; Ortiz de Montellano, Paul R.

    2008-01-01

    Heme oxygenase regiospecifically oxidizes heme at the α-meso position to give biliverdin IXα, CO, and iron. The heme orientation within the active site, which is thought to determine the oxidation regiospecificity, is shown here for the human enzyme (hHO1) to be largely determined by interactions between the heme carboxylic acid groups and residues Arg183 and Lys18 but not Tyr134. Mutation of either Arg183 or Lys18 individually does not significantly alter the NADPH-cytochrome P450 reductase-dependent reaction regiochemistry, but partially shifts the oxidation to the β/δ-meso positions in the reaction supported by ascorbic acid. Mutation of Glu29 to a lysine, which places a positive charge where it can interact with a heme carboxyl if the heme rotates by ~90°, causes a slight loss of regiospecificity, but combined with the R183E and K18E mutations results primarily in β/δ-meso oxidation of the heme under all conditions. NMR analysis of heme binding to the triple K18E/E29K/R183E mutant confirms rotation of the heme in the active site. Kinetic studies demonstrate that mutations of Arg183 greatly impair the rate of the P450 reductase-dependent reaction, in accord with the earlier finding that Arg183 is involved in binding of the reductase to hHO1, but have little effect on the ascorbate reaction. Mutations of Asp140 and Tyr58 that disrupt the active site hydrogen bonding network, impair catalytic rates but do not influence the oxidation regiochemistry. The results indicate both that the oxidation regiochemistry is largely controlled by ionic interactions of the heme propionic acid groups with the protein and that shifts in regiospecificity involve rotation of the heme about an axis perpendicular to the heme plane. PMID:16388581

  20. Heme isomers substantially affect heme's electronic structure and function

    DEFF Research Database (Denmark)

    Kepp, Kasper Planeta

    2017-01-01

    to similar energy of the isomers but with a sizable (25 kJ mol-1) barrier to interconversion arising from restricted rotation around the conjugated bonds. The four isomers, EE, EZ, ZE, and ZZ, were then investigated as 4-coordinate hemes, as 5-coordinate deoxyhemes, in 6-coordinate O2-adducts of globins.......e. the effects are not method-dependent. Thus, the nature of the isomer state is an important but overlooked feature of heme chemistry and function, and previous and future studies of hemes may be reconsidered in this new context....

  1. In vivo and in vitro olefin cyclopropanation catalyzed by heme enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Coelho, Pedro S.; Brustad, Eric M.; Arnold, Frances H.; Wang, Zhan; Lewis, Jared C.

    2016-11-15

    The present invention provides methods for catalyzing the conversion of an olefin to any compound containing one or more cyclopropane functional groups using heme enzymes. In certain aspects, the present invention provides a method for producing a cyclopropanation product comprising providing an olefinic substrate, a diazo reagent, and a heme enzyme; and admixing the components in a reaction for a time sufficient to produce a cyclopropanation product. In other aspects, the present invention provides heme enzymes including variants and fragments thereof that are capable of carrying out in vivo and in vitro olefin cyclopropanation reactions. Expression vectors and host cells expressing the heme enzymes are also provided by the present invention.

  2. In vivo and in vitro olefin cyclopropanation catalyzed by heme enzymes

    Science.gov (United States)

    Coelho, Pedro S; Brustad, Eric M; Arnold, Frances H; Wang, Zhan; Lewis, Jared C

    2015-03-31

    The present invention provides methods for catalyzing the conversion of an olefin to any compound containing one or more cyclopropane functional groups using heme enzymes. In certain aspects, the present invention provides a method for producing a cyclopropanation product comprising providing an olefinic substrate, a diazo reagent, and a heme enzyme; and admixing the components in a reaction for a time sufficient to produce a cyclopropanation product. In other aspects, the present invention provides heme enzymes including variants and fragments thereof that are capable of carrying out in vivo and in vitro olefin cyclopropanation reactions. Expression vectors and host cells expressing the heme enzymes are also provided by the present invention.

  3. Effect of a heme oxygenase-1 inducer on NADPH oxidase ...

    African Journals Online (AJOL)

    Purpose: To investigated the effect of hemin, a heme oxygenase-1 (HO-1) inducer, on nicotinamide adenine dinucleotide phosphate oxidase (NOX) expression in rats with alcohol-induced liver injury. Methods: Male Wistar rats were randomly divided into four groups consisting of the control group, the ethanol (EtOH) group, ...

  4. Structural Characterization of Heme Environmental Mutants of CgHmuT that Shuttles Heme Molecules to Heme Transporters

    Directory of Open Access Journals (Sweden)

    Norifumi Muraki

    2016-05-01

    Full Text Available Corynebacteria contain a heme uptake system encoded in hmuTUV genes, in which HmuT protein acts as a heme binding protein to transport heme to the cognate transporter HmuUV. The crystal structure of HmuT from Corynebacterium glutamicum (CgHmuT reveals that heme is accommodated in the central cleft with His141 and Tyr240 as the axial ligands and that Tyr240 forms a hydrogen bond with Arg242. In this work, the crystal structures of H141A, Y240A, and R242A mutants were determined to understand the role of these residues for the heme binding of CgHmuT. Overall and heme environmental structures of these mutants were similar to those of the wild type, suggesting that there is little conformational change in the heme-binding cleft during heme transport reaction with binding and the dissociation of heme. A loss of one axial ligand or the hydrogen bonding interaction with Tyr240 resulted in an increase in the redox potential of the heme for CgHmuT to be reduced by dithionite, though the wild type was not reduced under physiological conditions. These results suggest that the heme environmental structure stabilizes the ferric heme binding in CgHmuT, which will be responsible for efficient heme uptake under aerobic conditions where Corynebacteria grow.

  5. Giardia intestinalis incorporates heme into cytosolic cytochrome b₅.

    Science.gov (United States)

    Pyrih, Jan; Harant, Karel; Martincová, Eva; Sutak, Robert; Lesuisse, Emmanuel; Hrdý, Ivan; Tachezy, Jan

    2014-02-01

    The anaerobic intestinal pathogen Giardia intestinalis does not possess enzymes for heme synthesis, and it also lacks the typical set of hemoproteins that are involved in mitochondrial respiration and cellular oxygen stress management. Nevertheless, G. intestinalis may require heme for the function of particular hemoproteins, such as cytochrome b5 (cytb5). We have analyzed the sequences of eukaryotic cytb5 proteins and identified three distinct cytb5 groups: group I, which consists of C-tail membrane-anchored cytb5 proteins; group II, which includes soluble cytb5 proteins; and group III, which comprises the fungal cytb5 proteins. The majority of eukaryotes possess both group I and II cytb5 proteins, whereas three Giardia paralogs belong to group II. We have identified a fourth Giardia cytb5 paralog (gCYTb5-IV) that is rather divergent and possesses an unusual 134-residue N-terminal extension. Recombinant Giardia cytb5 proteins, including gCYTb5-IV, were expressed in Escherichia coli and exhibited characteristic UV-visible spectra that corresponded to heme-loaded cytb5 proteins. The expression of the recombinant gCYTb5-IV in G. intestinalis resulted in the increased import of extracellular heme and its incorporation into the protein, whereas this effect was not observed when gCYTb5-IV containing a mutated heme-binding site was expressed. The electrons for Giardia cytb5 proteins may be provided by the NADPH-dependent Tah18-like oxidoreductase GiOR-1. Therefore, GiOR-1 and cytb5 may constitute a novel redox system in G. intestinalis. To our knowledge, G. intestinalis is the first anaerobic eukaryote in which the presence of heme has been directly demonstrated.

  6. Rotationally Actuated Prosthetic Hand

    Science.gov (United States)

    Norton, William E.; Belcher, Jewell G., Jr.; Carden, James R.; Vest, Thomas W.

    1991-01-01

    Prosthetic hand attached to end of remaining part of forearm and to upper arm just above elbow. Pincerlike fingers pushed apart to degree depending on rotation of forearm. Simpler in design, simpler to operate, weighs less, and takes up less space.

  7. Dietary iron controls circadian hepatic glucose metabolism through heme synthesis.

    Science.gov (United States)

    Simcox, Judith A; Mitchell, Thomas Creighton; Gao, Yan; Just, Steven F; Cooksey, Robert; Cox, James; Ajioka, Richard; Jones, Deborah; Lee, Soh-Hyun; King, Daniel; Huang, Jingyu; McClain, Donald A

    2015-04-01

    The circadian rhythm of the liver maintains glucose homeostasis, and disruption of this rhythm is associated with type 2 diabetes. Feeding is one factor that sets the circadian clock in peripheral tissues, but relatively little is known about the role of specific dietary components in that regard. We assessed the effects of dietary iron on circadian gluconeogenesis. Dietary iron affects circadian glucose metabolism through heme-mediated regulation of the interaction of nuclear receptor subfamily 1 group d member 1 (Rev-Erbα) with its cosuppressor nuclear receptor corepressor 1 (NCOR). Loss of regulated heme synthesis was achieved by aminolevulinic acid (ALA) treatment of mice or cultured cells to bypass the rate-limiting enzyme in hepatic heme synthesis, ALA synthase 1 (ALAS1). ALA treatment abolishes differences in hepatic glucose production and in the expression of gluconeogenic enzymes seen with variation of dietary iron. The differences among diets are also lost with inhibition of heme synthesis with isonicotinylhydrazine. Dietary iron modulates levels of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), a transcriptional activator of ALAS1, to affect hepatic heme. Treatment of mice with the antioxidant N-acetylcysteine diminishes PGC-1α variation observed among the iron diets, suggesting that iron is acting through reactive oxygen species signaling. © 2015 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered.

  8. Cortical neural prosthetics.

    Science.gov (United States)

    Schwartz, Andrew B

    2004-01-01

    Control of prostheses using cortical signals is based on three elements: chronic microelectrode arrays, extraction algorithms, and prosthetic effectors. Arrays of microelectrodes are permanently implanted in cerebral cortex. These arrays must record populations of single- and multiunit activity indefinitely. Information containing position and velocity correlates of animate movement needs to be extracted continuously in real time from the recorded activity. Prosthetic arms, the current effectors used in this work, need to have the agility and configuration of natural arms. Demonstrations using closed-loop control show that subjects change their neural activity to improve performance with these devices. Adaptive-learning algorithms that capitalize on these improvements show that this technology has the capability of restoring much of the arm movement lost with immobilizing deficits.

  9. Welding of Prosthetic Alloys

    Directory of Open Access Journals (Sweden)

    Wojciechowska M.

    2015-04-01

    Full Text Available This paper presents the techniques of joining metal denture elements, used in prosthetic dentistry: the traditional soldering technique with a gas burner and a new technique of welding with a laser beam; the aim of the study was to make a comparative assessment of the quality of the joints in view of the possibility of applying them in prosthetic structures. Fractographic examinations were conducted along with tensile strength and impact strength tests, and the quality of the joints was assessed compared to the solid metal. The experiments have shown that the metal elements used to make dentures, joined by the technique which employs a laser beam, have better strength properties than those achieved with a gas burner.

  10. Heme oxygenase-1, oxidation, inflammation and atherosclerosis

    Directory of Open Access Journals (Sweden)

    Jesus A Araujo

    2012-07-01

    Full Text Available Atherosclerosis is an inflammatory process of the vascular wall characterized by the infiltration of lipids and inflammatory cells. Oxidative modifications of infiltrating low density lipoproteins and induction of oxidative stress play a major role in lipid retention in the vascular wall, uptake by macrophages and generation of foam cells, a hallmark of this disorder. The vasculature has a plethora of protective resources against oxidation and inflammation, many of them regulated by the Nrf2 transcription factor. Heme oxygenase-1 (HO-1 is a Nrf2-regulated gene that plays a critical role in the prevention of vascular inflammation. It is the inducible isoform of heme oxygenase, responsible for the oxidative cleavage of heme groups leading to the generation of biliverdin, carbon monoxide and release of ferrous iron. HO-1 has important antioxidant, antiinflammatory, antiapoptotic, antiproliferative and immunomodulatory effects in vascular cells, most of which play a significant role in the protection against atherogenesis. HO-1 may also be an important feature in macrophage differentiation and polarization to certain subtypes. The biological effects of HO-1 are largely attributable to its enzymatic activity, which can be conceived as a system with three arms of action, corresponding to its three enzymatic byproducts. HO-1 mediated vascular protection may be due to a combination of systemic and vascular local effects. It is usually expressed at low levels but can be highly upregulated in the presence of several proatherogenic stimuli. The HO-1 system is amenable for use in the development of new therapies, some of them currently under experimental and clinical trials. Interestingly, in contrast to the HO-1 antiatherogenic actions, the expression of its transcriptional regulator Nrf2 leads to proatherogenic effects instead. This article reviews the evidence that supports the antiatherogenic role of HO-1, potential pathways and mechanisms mediating

  11. Resonance Raman and EPR spectroscopic studies on heme-heme oxygenase complexes.

    Science.gov (United States)

    Sun, J; Wilks, A; Ortiz de Montellano, P R; Loehr, T M

    1993-12-28

    The binding of ferrous and ferric hemes and manganese(II)- and manganese(III)-substituted hemes to heme oxygenase has been investigated by optical absorption, resonance Raman, and EPR spectroscopy. The results are consistent with the presence of a six-coordinate heme moiety ligated to an essential histidine ligand and a water molecule. The latter ionizes with a pKa approximately 8.0 to give a mixture of high-spin and low-spin six-coordinate hydroxo adducts. Addition of excess cyanide converts the heme to a hexacoordinate low-spin species. The resonance Raman spectrum of the ferrous heme-heme oxygenase complex and that of the Mn(II)protoporphyrin-heme oxygenase complex shows bands at 216 and 212 cm-1, respectively, that are assigned to the metal-histidine stretching mode. The EPR spectrum of the oxidized heme-heme oxygenase complex has a strongly axial signal with g parallel of approximately 6 and g perpendicular approximately 2. 14NO and 15NO adducts of ferrous heme-heme oxygenase exhibit EPR hyperfine splittings of approximately 20 and approximately 25 Gauss, respectively. In addition, both nitrosyl complexes show additional superhyperfine splittings of approximately 7 Gauss from spin-spin interaction with the proximal histidine nitrogen. The heme environment in the heme-heme oxygenase enzyme-substrate complex has spectroscopic properties similar to those of the heme in myoglobin. Hence, there is neither a strongly electron-donating fifth (proximal) ligand nor an electron-withdrawing network on the distal side of the heme moiety comparable to that for cytochromes P-450 and peroxidases. This observation has profound implications about the nature of the oxygen-activating process in the heme-->biliverdin reaction that are discussed in this paper.

  12. Prosthetic elbow joint

    Science.gov (United States)

    Weddendorf, Bruce C. (Inventor)

    1994-01-01

    An artificial, manually positionable elbow joint for use in an upper extremity, above-elbow, prosthetic is described. The prosthesis provides a locking feature that is easily controlled by the wearer. The instant elbow joint is very strong and durable enough to withstand the repeated heavy loadings encountered by a wearer who works in an industrial, construction, farming, or similar environment. The elbow joint of the present invention comprises a turntable, a frame, a forearm, and a locking assembly. The frame generally includes a housing for the locking assembly and two protruding ears. The forearm includes an elongated beam having a cup-shaped cylindrical member at one end and a locking wheel having a plurality of holes along a circular arc on its other end with a central bore for pivotal attachment to the protruding ears of the frame. The locking assembly includes a collar having a central opening with a plurality of internal grooves, a plurality of internal cam members each having a chamfered surface at one end and a V-shaped slot at its other end; an elongated locking pin having a crown wheel with cam surfaces and locking lugs secured thereto; two coiled compression springs; and a flexible filament attached to one end of the elongated locking pin and extending from the locking assembly for extending and retracting the locking pin into the holes in the locking wheel to permit selective adjustment of the forearm relative to the frame. In use, the turntable is affixed to the upper arm part of the prosthetic in the conventional manner, and the cup-shaped cylindrical member on one end of the forearm is affixed to the forearm piece of the prosthetic in the conventional manner. The elbow joint is easily adjusted and locked between maximum flex and extended positions.

  13. Passive prosthetic hands and tools: A literature review.

    Science.gov (United States)

    Maat, Bartjan; Smit, Gerwin; Plettenburg, Dick; Breedveld, Paul

    2018-02-01

    The group of passive prostheses consists of prosthetic hands and prosthetic tools. These can either be static or adjustable. Limited research and development on passive prostheses has been performed although many people use these prosthesis types. Although some publications describe passive prostheses, no recent review of the peer-reviewed literature on passive prostheses is available. Review the peer-reviewed literature on passive prostheses for replacement of the hand. Literature review. Four electronic databases were searched using a Boolean combination of relevant keywords. English-language articles relevant to the objective were selected. In all, 38 papers were included in the review. Publications on passive prosthetic hands describe their users, usage, functionality, and problems in activities of daily living. Publications on prosthetic tools mostly focus on sport, recreation, and vehicle driving. Passive hand prostheses receive little attention in prosthetic research and literature. Yet one out of three people with a limb deficiency uses this type of prosthesis. Literature indicates that passive prostheses can be improved on pulling and grasping functions. In the literature, ambiguous names are used for different types of passive prostheses. This causes confusion. We present a new and clear classification of passive prostheses. Clinical relevance This review provides information on the users of passive prosthetic hands and tools, their usage and the functionality. Passive prostheses receive very little attention and low appreciation in literature. Passive prosthetic hands and tools show to be useful to many unilateral amputees and should receive more attention and higher acceptance.

  14. Structural and biochemical characterization of two heme binding sites on α1-microglobulin using site directed mutagenesis and molecular simulation.

    Science.gov (United States)

    Rutardottir, Sigurbjörg; Karnaukhova, Elena; Nantasenamat, Chanin; Songtawee, Napat; Prachayasittikul, Virapong; Rajabi, Mohsen; Rosenlöf, Lena Wester; Alayash, Abdu I; Åkerström, Bo

    2016-01-01

    α1-Microglobulin (A1M) is a reductase and radical scavenger involved in physiological protection against oxidative damage. These functions were previously shown to be dependent upon cysteinyl-, C34, and lysyl side-chains, K(92, 118,130). A1M binds heme and the crystal structure suggests that C34 and H123 participate in a heme binding site. We have investigated the involvement of these five residues in the interactions with heme. Four A1M-variants were expressed: with cysteine to serine substitution in position 34, lysine to threonine substitutions in positions (92, 118, 130), histidine to serine substitution in position 123 and a wt without mutations. Heme binding was investigated by tryptophan fluorescence quenching, UV-Vis spectrophotometry, circular dichroism, SPR, electrophoretic migration shift, gel filtration, catalase-like activity and molecular simulation. All A1M-variants bound to heme. Mutations in C34, H123 or K(92, 118, 130) resulted in significant absorbance changes, CD spectral changes, and catalase-like activity, suggesting involvement of these side-groups in coordination of the heme-iron. Molecular simulation support a model with two heme-binding sites in A1M involving the mutated residues. Binding of the first heme induces allosteric stabilization of the structure predisposing for a better fit of the second heme. The results suggest that one heme-binding site is located in the lipocalin pocket and a second binding site between loops 1 and 4. Reactions with the hemes involve the side-groups of C34, K(92, 118, 130) and H123. The model provides a structural basis for the functional activities of A1M: heme binding activity of A1M. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. The 2-Cys Peroxiredoxin Alkyl Hydroperoxide Reductase C Binds Heme and Participates in Its Intracellular Availability in Streptococcus agalactiae*

    Science.gov (United States)

    Lechardeur, Delphine; Fernandez, Annabelle; Robert, Bruno; Gaudu, Philippe; Trieu-Cuot, Patrick; Lamberet, Gilles; Gruss, Alexandra

    2010-01-01

    Heme is a redox-reactive molecule with vital and complex roles in bacterial metabolism, survival, and virulence. However, few intracellular heme partners were identified to date and are not well conserved in bacteria. The opportunistic pathogen Streptococcus agalactiae (group B Streptococcus) is a heme auxotroph, which acquires exogenous heme to activate an aerobic respiratory chain. We identified the alkyl hydroperoxide reductase AhpC, a member of the highly conserved thiol-dependent 2-Cys peroxiredoxins, as a heme-binding protein. AhpC binds hemin with a Kd of 0.5 μm and a 1:1 stoichiometry. Mutagenesis of cysteines revealed that hemin binding is dissociable from catalytic activity and multimerization. AhpC reductase activity was unchanged upon interaction with heme in vitro and in vivo. A group B Streptococcus ahpC mutant displayed attenuation of two heme-dependent functions, respiration and activity of a heterologous catalase, suggesting a role for AhpC in heme intracellular fate. In support of this hypothesis, AhpC-bound hemin was protected from chemical degradation in vitro. Our results reveal for the first time a role for AhpC as a heme-binding protein. PMID:20332091

  16. [Hereditary porphyrias and heme related disorders].

    Science.gov (United States)

    Puy, Hervé; Gouya, Laurent; Deybach, Jean-charles

    2014-06-01

    Hereditary porphyrias comprise a group of eight metabolic disorders of the haem biosynthesis pathway, characterised by acute neurovisceral symptoms and/or skin lesions. Each porphyria is caused by abnormal functioning of a particular enzymatic step, resulting in specific accumulation of heme precursors. Seven porphyrias are due to a partial enzyme deficiency, while a gain-of-function mechanism has recently been identify in a novel porphyria. Acute porphyrias present with severe abdominal pain, nausea, constipation and confusion, and are sometimes complicated by seizures and severe neurological disorders, which may be life-threatening. Cutaneous porphyrias can also be present, with either acute painful photosensitivity or skin fragility and blisters. Rare recessive porphyrias usually manifest in early childhood with either severe chronic neurological symptoms or chronic haemolysis and severe cutaneous photosensitivity. Porphyrias are still under-diagnosed, but recent advances in the pathogenesis and genetics of human porphyrias are leading to better care of these patients and their families.

  17. New developments in prosthetic arm systems

    Directory of Open Access Journals (Sweden)

    Vujaklija I

    2016-07-01

    Full Text Available Ivan Vujaklija,1 Dario Farina,1 Oskar C Aszmann2 1Institute of Neurorehabilitation Systems, Bernstein Focus Neurotechnology Göttingen, University Medical Center Göttingen, Georg-August University, Göttingen, Germany; 2Christian Doppler Laboratory for Restoration of Extremity Function, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Vienna, Austria Abstract: Absence of an upper limb leads to severe impairments in everyday life, which can further influence the social and mental state. For these reasons, early developments in cosmetic and body-driven prostheses date some centuries ago, and they have been evolving ever since. Following the end of the Second World War, rapid developments in technology resulted in powered myoelectric hand prosthetics. In the years to come, these devices were common on the market, though they still suffered high user abandonment rates. The reasons for rejection were trifold – insufficient functionality of the hardware, fragile design, and cumbersome control. In the last decade, both academia and industry have reached major improvements concerning technical features of upper limb prosthetics and methods for their interfacing and control. Advanced robotic hands are offered by several vendors and research groups, with a variety of active and passive wrist options that can be articulated across several degrees of freedom. Nowadays, elbow joint designs include active solutions with different weight and power options. Control features are getting progressively more sophisticated, offering options for multiple sensor integration and multi-joint articulation. Latest developments in socket designs are capable of facilitating implantable and multiple surface electromyography sensors in both traditional and osseointegration-based systems. Novel surgical techniques in combination with modern, sophisticated hardware are enabling restoration of dexterous upper limb

  18. Developing prescribing guidelines for microprocessor-controlled prosthetic knees in the South East England.

    Science.gov (United States)

    Sedki, Imad; Fisher, Keren

    2015-06-01

    Microprocessor-controlled prosthetic knees have gained increasing popularity over the last decade. Research supports their provision to address specific problems or to achieve certain rehabilitation goals. However, there are yet no agreed protocols or prescribing criteria to assist clinicians in the identification and appropriate selection of suitable users. The aim is to reach professionals' agreement on specific prescribing guidelines for microprocessor-controlled prosthetic knees. The study involved multidisciplinary teams from the Inter Regional Prosthetic Audit Group, representing nine Prosthetic Rehabilitation Centres in the South East England region. We used the Delphi technique with a total of three rounds to reach professionals' agreement. The prescribing guidelines were agreed and will be reviewed and updated depending on new research evidence and technical advances. This project is highly useful for professionals in a clinic setting to aid in appropriate patient selection and to justify the cost of prescribing microprocessor-controlled prosthetic knees. © The International Society for Prosthetics and Orthotics 2014.

  19. The surface protein Shr of Streptococcus pyogenes binds heme and transfers it to the streptococcal heme-binding protein Shp

    Directory of Open Access Journals (Sweden)

    Lei Benfang

    2008-01-01

    Full Text Available Abstract Background The heme acquisition machinery in Streptococcus pyogenes is believed to consist of the surface proteins, Shr and Shp, and heme-specific ATP-binding cassette transporter HtsABC. Shp has been shown to rapidly transfer its heme to the lipoprotein component, HtsA, of HtsABC. The function of Shr and the heme source of Shp have not been established. Results The objective of this study was to determine whether Shr binds heme and is a heme source of Shp. To achieve the objective, recombinant Shr protein was prepared. The purified Shr displays a spectrum typical of hemoproteins, indicating that Shr binds heme and acquires heme from Escherichia coli hemoproteins in vivo. Spectral analysis of Shr and Shp isolated from a mixture of Shr and heme-free Shp (apoShp indicates that Shr and apoShp lost and gained heme, respectively; whereas Shr did not efficiently lose its heme in incubation with apoHtsA under the identical conditions. These results suggest that Shr directly transfers its heme to Shp. In addition, the rates of heme transfer from human hemoglobin to apoShp are close to those of simple ferric heme dissociation from hemoglobin, suggesting that methemoglobin does not directly transfer its heme to apoShp. Conclusion We have demonstrated that recombinant Shr can acquire heme from E. coli hemoproteins in vivo and appears to directly transfer its heme to Shp and that Shp appears not to directly acquire heme from human methemoglobin. These results suggest the possibility that Shr is a source of heme for Shp and that the Shr-to-Shp heme transfer is a step of the heme acquisition process in S. pyogenes. Further characterization of the Shr/Shp/HtsA system would advance our understanding of the mechanism of heme acquisition in S. pyogenes.

  20. Prosthetic helping hand

    Science.gov (United States)

    Vest, Thomas W. (Inventor); Carden, James R. (Inventor); Norton, William E. (Inventor); Belcher, Jewell G. (Inventor)

    1992-01-01

    A prosthetic device for below-the-elbow amputees, having a C-shaped clamping mechanism for grasping cylindrical objects, is described. The clamping mechanism is pivotally mounted to a cuff that fits on the amputee's lower arm. The present invention is utilized by placing an arm that has been amputated below the elbow into the cuff. The clamping mechanism then serves as a hand whenever it becomes necessary for the amputee to grasp a cylindrical object such as a handle, a bar, a rod, etc. To grasp the cylindrical object, the object is jammed against the opening in the C-shaped spring, causing the spring to open, the object to pass to the center of the spring, and the spring to snap shut behind the object. Various sizes of clamping mechanisms can be provided and easily interchanged to accommodate a variety of diameters. With the extension that pivots and rotates, the clamping mechanism can be used in a variety of orientations. Thus, this invention provides the amputee with a clamping mechanism that can be used to perform a number of tasks.

  1. Dioxygen reactivity of meso-hydroxylated hemes: intermediates in ...

    Indian Academy of Sciences (India)

    Keywords. Heme oxygenase; heme degradation; coupled oxidation; variable temperature paramagnetic NMR. Abstract. Heme oxygenase (HO) is the only enzyme in mammals known to catalyse the physiological degradation of unwanted heme into biliverdin, Fe ion and CO. The process involves introduction of the hydroxyl ...

  2. Protein Machineries Involved in the Attachment of Heme to Cytochrome c: Protein Structures and Molecular Mechanisms

    Directory of Open Access Journals (Sweden)

    Carlo Travaglini-Allocatelli

    2013-01-01

    Full Text Available Cytochromes c (Cyt c are ubiquitous heme-containing proteins, mainly involved in electron transfer processes, whose structure and functions have been and still are intensely studied. Surprisingly, our understanding of the molecular mechanism whereby the heme group is covalently attached to the apoprotein (apoCyt in the cell is still largely unknown. This posttranslational process, known as Cyt c biogenesis or Cyt c maturation, ensures the stereospecific formation of the thioether bonds between the heme vinyl groups and the cysteine thiols of the apoCyt heme binding motif. To accomplish this task, prokaryotic and eukaryotic cells have evolved distinctive protein machineries composed of different proteins. In this review, the structural and functional properties of the main maturation apparatuses found in gram-negative and gram-positive bacteria and in the mitochondria of eukaryotic cells will be presented, dissecting the Cyt c maturation process into three functional steps: (i heme translocation and delivery, (ii apoCyt thioreductive pathway, and (iii apoCyt chaperoning and heme ligation. Moreover, current hypotheses and open questions about the molecular mechanisms of each of the three steps will be discussed, with special attention to System I, the maturation apparatus found in gram-negative bacteria.

  3. Clinical Aspects of Combination of Aesthetic Fixed Prosthetic Appliances

    Directory of Open Access Journals (Sweden)

    Andrii Biben

    2017-10-01

    Full Text Available The objective of the research was to study the peculiarities of the combination of zirconia and ceramic occlusal surfaces when constructing aesthetic fixed prosthetic appliances. Materials and methods. The study included 70 patients with zirconia and ceramic occlusal surfaces of aesthetic fixed dental prostheses. Group I included 24 patients with a combination of zirconia and ceramic occlusal surfaces. Group II included 30 patients with a combination of ceramic occlusal surfaces. Group III included 16 patients with a combination of zirconia occlusal surfaces. All the patients were observed 12 and 24 months after prosthetic repair. Results. 12 and 24 months after prosthetic repair, the occlusal contact surface area was the largest in Group II (8.18±0.16 mm2 and 9.17±0.1 mm2, respectively. In Group I, where only one occlusive surface was made of zirconium dioxide, significantly reduced levels of abrasion were observed as compared to Group II – 8.07±0.21 mm2 and 8.65±0.23 mm2, respectively. 36 months after denture wearing, in Group III, the smallest contact surface area – 7.84±0.15 mm2as well as the lowest growth of the surface area was observed – 8.07±0.13 mm2. Conclusions. Dental prostheses with at least one ceramic occlusal surface exhibit a strong tendency to abrasion and, consequently, to an increase in the occlusal surface area resulting in an excessive load on prosthetic appliance. Moreover, functional and aesthetic values of prosthetic prosthesis sharply decrease. Therefore, we recommend to produce zirconia occlusal surface or at least to combine the same materials, as it will increase the longevity of prosthetic appliance.

  4. Heme environment in HmuY, the heme-binding protein of Porphyromonas gingivalis

    Energy Technology Data Exchange (ETDEWEB)

    Wojtowicz, Halina [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland); Wojaczynski, Jacek [Department of Chemistry, University of Wroclaw, 50-383 Wroclaw (Poland); Olczak, Mariusz [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland); Kroliczewski, Jaroslaw [Laboratory of Biophysics, Faculty of Biotechnology, University of Wroclaw, 50-148 Wroclaw (Poland); Latos-Grazynski, Lechoslaw [Department of Chemistry, University of Wroclaw, 50-383 Wroclaw (Poland); Olczak, Teresa, E-mail: Teresa.Olczak@biotech.uni.wroc.pl [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland)

    2009-05-29

    Porphyromonas gingivalis, a Gram-negative anaerobic bacterium implicated in the development and progression of chronic periodontitis, acquires heme for growth by a novel mechanism composed of HmuY and HmuR proteins. The aim of this study was to characterize the nature of heme binding to HmuY. The protein was expressed, purified and detailed investigations using UV-vis absorption, CD, MCD, and {sup 1}H NMR spectroscopy were carried out. Ferric heme bound to HmuY may be reduced by sodium dithionite and re-oxidized by potassium ferricyanide. Heme complexed to HmuY, with a midpoint potential of 136 mV, is in a low-spin Fe(III) hexa-coordinate environment. Analysis of heme binding to several single and double HmuY mutants with the methionine, histidine, cysteine, or tyrosine residues replaced by an alanine residue identified histidines 134 and 166 as potential heme ligands.

  5. Challenging Density Functional Theory Calculations with Hemes and Porphyrins

    Directory of Open Access Journals (Sweden)

    Sam P. de Visser

    2016-04-01

    Full Text Available In this paper we review recent advances in computational chemistry and specifically focus on the chemical description of heme proteins and synthetic porphyrins that act as both mimics of natural processes and technological uses. These are challenging biochemical systems involved in electron transfer as well as biocatalysis processes. In recent years computational tools have improved considerably and now can reproduce experimental spectroscopic and reactivity studies within a reasonable error margin (several kcal·mol−1. This paper gives recent examples from our groups, where we investigated heme and synthetic metal-porphyrin systems. The four case studies highlight how computational modelling can correctly reproduce experimental product distributions, predicted reactivity trends and guide interpretation of electronic structures of complex systems. The case studies focus on the calculations of a variety of spectroscopic features of porphyrins and show how computational modelling gives important insight that explains the experimental spectra and can lead to the design of porphyrins with tuned properties.

  6. Using heme as an energy boost for lactic acid bacteria.

    Science.gov (United States)

    Lechardeur, Delphine; Cesselin, Bénédicte; Fernandez, Annabelle; Lamberet, Gilles; Garrigues, Christel; Pedersen, Martin; Gaudu, Philippe; Gruss, Alexandra

    2011-04-01

    Lactic acid bacteria (LAB) are a phylogenetically diverse group named for their main attribute in food fermentations, that is, production of lactic acid. However, several LAB are genetically equipped for aerobic respiration metabolism when provided with exogenous sources of heme (and menaquinones for some species). Respiration metabolism is energetically favorable and leads to less oxidative and acid stress during growth. As a consequence, the growth and survival of several LAB can be dramatically improved under respiration-permissive conditions. Respiration metabolism already has industrial applications for the production of dairy starter cultures. In view of the growth and survival advantages conferred by respiration, and the availability of heme and menaquinones in natural environments, we recommend that respiration be accepted as a part of the natural lifestyle of numerous LAB. Copyright © 2010 Elsevier Ltd. All rights reserved.

  7. Challenging Density Functional Theory Calculations with Hemes and Porphyrins

    Science.gov (United States)

    de Visser, Sam P.; Stillman, Martin J.

    2016-01-01

    In this paper we review recent advances in computational chemistry and specifically focus on the chemical description of heme proteins and synthetic porphyrins that act as both mimics of natural processes and technological uses. These are challenging biochemical systems involved in electron transfer as well as biocatalysis processes. In recent years computational tools have improved considerably and now can reproduce experimental spectroscopic and reactivity studies within a reasonable error margin (several kcal·mol−1). This paper gives recent examples from our groups, where we investigated heme and synthetic metal-porphyrin systems. The four case studies highlight how computational modelling can correctly reproduce experimental product distributions, predicted reactivity trends and guide interpretation of electronic structures of complex systems. The case studies focus on the calculations of a variety of spectroscopic features of porphyrins and show how computational modelling gives important insight that explains the experimental spectra and can lead to the design of porphyrins with tuned properties. PMID:27070578

  8. Alterations in the heme biosynthetic pathway as an index of exposure to toxins

    Energy Technology Data Exchange (ETDEWEB)

    Marks, G.S.; Zelt, D.T.; Cole, S.P.

    1982-07-01

    Under normal circumstances the heme biosynthetic pathway is carefully controlled and porphyrins are formed in only trace amounts. When control mechanisms are disturbed by xenobiotics, porphyrins may be formed and serve as a signal of the interaction between a xenobiotic and the heme biosynthetic pathway. For example, porphyrinuria was an early manifestation of a hexachlorobenzene-induced porphyria outbreak in Turkey. In humans exposed to polybrominated biphenyls and to 2,3,7,8-tetrachlorodibenzo-p-dioxin the urinary porphyrin pattern was significantly different from normal in a large number of exposed individuals. The question is raised whether measurement of urinary porphyrin profiles by improved methods will enable an estimate to be made of the extent of exposure to haloaromatic hydrocarbons in the human population. A wide variety of xenobiotics interact with the prosthetic heme of cytochrome P-450 forming novel N-alkylporphyrins. Identification of these N-alkylporphyrins in body fluids might provide a means of assessing exposure to a variety of xenobiotics in human populations.

  9. Crystal structure of rat heme oxygenase-1 in complex with biliverdin-iron chelate. Conformational change of the distal helix during the heme cleavage reaction.

    Science.gov (United States)

    Sugishima, Masakazu; Sakamoto, Hiroshi; Higashimoto, Yuichiro; Noguchi, Masato; Fukuyama, Keiichi

    2003-08-22

    The crystal structure of rat heme oxygenase-1 in complex with biliverdin-iron chelate (biliverdin(Fe)-HO-1), the immediate precursor of the final product, biliverdin, has been determined at a 2.4-A resolution. The electron density in the heme pocket clearly showed that the tetrapyrrole ring of heme is cleaved at the alpha-meso edge. Like the heme bound to HO-1, biliverdin-iron chelate is located between the distal and proximal helices, but its accommodation state seems to be less stable in light of the disordering of the solvent-exposed propionate and vinyl groups. The middle of the distal helix is shifted away from the center of the active site in biliverdin(Fe)-HO-1, increasing the size of the heme pocket. The hydrogen-bonding interaction between Glu-29 and Gln-38, considered to restrain the orientation of the proximal helix in the heme-HO-1 complex, was lost in biliverdin(Fe)-HO-1, leading to relaxation of the helix. Biliverdin has a distorted helical conformation; the lactam oxygen atom of its pyrrole ring-A interacted with Asp-140 through a hydrogen-bonding solvent network. Because of the absence of a distal water ligand, the iron atom is five-coordinated with His-25 and four pyrrole nitrogen atoms. The coordination geometry deviates considerably from a square pyramid, suggesting that the iron may be readily dissociated. We speculate that the opened conformation of the heme pocket facilitates sequential product release, first iron then biliverdin, and that because of biliverdin's increased flexibility, iron release triggers its slow dissociation.

  10. In vitro Activation of heme oxygenase-2 by menadione and its analogs.

    Science.gov (United States)

    Vukomanovic, Dragic; Rahman, Mona N; Bilokin, Yaroslav; Golub, Andriy G; Brien, James F; Szarek, Walter A; Jia, Zongchao; Nakatsu, Kanji

    2014-02-18

    Previously, we reported that menadione activated rat, native heme oxygenase-2 (HO-2) and human recombinant heme oxygenase-2 selectively; it did not activate spleen, microsomal heme oxygenase-1. The purpose of this study was to explore some structure-activity relationships of this activation and the idea that redox properties may be an important aspect of menadione efficacy. Heme oxygenase activity was determined in vitro using rat spleen and brain microsomes as the sources of heme oxygenase-1 and -2, respectively, as well as recombinant, human heme oxygenase-2. Menadione analogs with bulky aliphatic groups at position-3, namely vitamins K1 and K2, were not able to activate HO-2. In contrast, several compounds with similar bulky but less lipophilic moieties at position-2 (and -3) were able to activate HO-2 many fold; these compounds included polar, rigid, furan-containing naphthoquinones, furan-benzoxazine naphthoquinones, 2-(aminophenylphenyl)-3-piperidin-1-yl naphthoquinones. To explore the idea that redox properties might be involved in menadione efficacy, we tested analogs such as 1,4-dimethoxy-2-methylnaphthalene, pentafluoromenadione, monohalogenated naphthoquinones, α-tetralone and 1,4-naphthoquinone. All of these compounds were inactive except for 1,4-naphthoquinone. Menadione activated full-length recombinant human heme oxygenase-2 (FL-hHO-2) as effectively as rat brain enzyme, but it did not activate rat spleen heme oxygenase. These observations are consistent with the idea that naphthoquinones such as menadione bind to a receptor in HO-2 and activate the enzyme through a mechanism that may involve redox properties.

  11. Heme Compounds in Dinosaur Trabecular Bone

    National Research Council Canada - National Science Library

    Mary H. Schweitzer; Mark Marshall; Keith Carron; D. Scott Bohle; Scott C. Busse; Ernst V. Arnold; Darlene Barnard; J. R. Horner; Jean R. Starkey

    1997-01-01

    Six independent lines of evidence point to the existence of heme-containing compounds and/or hemoglobin breakdown products in extracts of trabecular tissues of the large theropod dinosaur Tyrannosaurus rex...

  12. Prescription and repair rates of prosthetic limbs in the VA healthcare system: implications for national prosthetic parity.

    Science.gov (United States)

    Etter, Katherine; Borgia, Matthew; Resnik, Linda

    2014-05-22

    Abstract Purpose: To quantify prescription and repair rates of prosthetic limbs in the Department of Veterans Affairs (VA) and explore differences by level, type, and age. Methods: Veterans (N = 32 440) with an initial prosthetic prescription between 2000 and 2010 were classified by amputation level and type. Annual rates of prescription and repair were calculated using person-time and compared by group. Results: Veterans with upper limb amputation had lower annual prescription and repair rates (0.28 and 0.21) compared with those with lower limb amputation (0.40 and 0.56). Myoelectric devices users had higher prescription rates. However, body-powered users had higher repair rates. Prescription and repair rates for microprocessor knee joints were higher than for fluid and friction devices. Veterans under 65 had 0.07 and 0.16 higher rates of prescription and repair than those over 65 (p < 0.0001). Conclusions: Because the VA is unconstrained by co-pays or caps, data on prosthetic prescription and repair can be used to estimate rates that might occur if national prosthetic parity laws were adopted. Given the rates found, it is likely that annual costs would exceed the typical annual and/or lifetime caps in most insurance plans. In states without prosthetic parity laws, such costs likely limit access to needed devices. Implications for Rehabilitation For the almost 2 million people in the United States living with an amputation or congenital limb loss, purchasing and maintaining a prosthetic limb can be costly, with insurances often imposing annual or lifetime caps. Data on prosthetic purchasing and repair is limited and reliant on self-reported information. Because the VA is unconstrained by co-pays or caps, claims data on prosthetic prescription and repair can be used to estimate rates that might occur if national prosthetic parity laws were adopted. Given the rates found, it is likely that annual costs would exceed the typical annual and/or lifetime caps

  13. Candida infection of a prosthetic shoulder joint

    Energy Technology Data Exchange (ETDEWEB)

    Lichtman, E.A.

    1983-09-01

    A heroin addict developed a Candida parapsilosis infection in a prosthetic shoulder joint. Radiographs showed loose fragments of cement with prosthetic loosening. The patient was treated with removal of the prosthesis and intravenous amphotericin B followed by oral ketoconazole.

  14. Rothia prosthetic knee joint infection.

    Science.gov (United States)

    Trivedi, Manish N; Malhotra, Prashant

    2015-08-01

    Rothia species - Gram-positive pleomorphic bacteria that are part of the normal oral and respiratory flora - are commonly associated with dental cavities and periodontal disease although systemic infections have been described. We describe a 53-year-old female with rheumatoid arthritis complicated by prosthetic knee joint infection due to Rothia species, which was successfully treated by surgical removal of prosthesis and prolonged antimicrobial therapy. The issue of antibiotic prophylaxis before dental procedures among patients with prosthetic joint replacements is discussed. Copyright © 2012. Published by Elsevier B.V.

  15. Structural mechanisms of nonplanar hemes in proteins

    Energy Technology Data Exchange (ETDEWEB)

    Shelnutt, J.A.

    1997-05-01

    The objective is to assess the occurrence of nonplanar distortions of hemes and other tetrapyrroles in proteins and to determine the biological function of these distortions. Recently, these distortions were found by us to be conserved among proteins belonging to a functional class. Conservation of the conformation of the heme indicates a possible functional role. Researchers have suggested possible mechanisms by which heme distortions might influence biological properties; however, no heme distortion has yet been shown conclusively to participate in a structural mechanism of hemoprotein function. The specific aims of the proposed work are: (1) to characterize and quantify the distortions of the hemes in all of the more than 300 hemoprotein X-ray crystal structures in terms of displacements along the lowest-frequency normal coordinates, (2) to determine the structural features of the protein component that generate and control these nonplanar distortions by using spectroscopic studies and molecular-mechanics calculations for the native proteins, their mutants and heme-peptide fragments, and model porphyrins, (3) to determine spectroscopic markers for the various types of distortion, and, finally, (4) to discover the functional significance of the nonplanar distortions by correlating function with porphyrin conformation for proteins and model porphyrins.

  16. Heme Oxygenases in Cardiovascular Health and Disease

    Science.gov (United States)

    Ayer, Anita; Zarjou, Abolfazl; Agarwal, Anupam; Stocker, Roland

    2016-01-01

    Heme oxygenases are composed of two isozymes, Hmox1 and Hmox2, that catalyze the degradation of heme to carbon monoxide (CO), ferrous iron, and biliverdin, the latter of which is subsequently converted to bilirubin. While initially considered to be waste products, CO and biliverdin/bilirubin have been shown over the last 20 years to modulate key cellular processes, such as inflammation, cell proliferation, and apoptosis, as well as antioxidant defense. This shift in paradigm has led to the importance of heme oxygenases and their products in cell physiology now being well accepted. The identification of the two human cases thus far of heme oxygenase deficiency and the generation of mice deficient in Hmox1 or Hmox2 have reiterated a role for these enzymes in both normal cell function and disease pathogenesis, especially in the context of cardiovascular disease. This review covers the current knowledge on the function of both Hmox1 and Hmox2 at both a cellular and tissue level in the cardiovascular system. Initially, the roles of heme oxygenases in vascular health and the regulation of processes central to vascular diseases are outlined, followed by an evaluation of the role(s) of Hmox1 and Hmox2 in various diseases such as atherosclerosis, intimal hyperplasia, myocardial infarction, and angiogenesis. Finally, the therapeutic potential of heme oxygenases and their products are examined in a cardiovascular disease context, with a focus on how the knowledge we have gained on these enzymes may be capitalized in future clinical studies. PMID:27604527

  17. Unique structure and stability of HmuY, a novel heme-binding protein of Porphyromonas gingivalis.

    Directory of Open Access Journals (Sweden)

    Halina Wójtowicz

    2009-05-01

    Full Text Available Infection, survival, and proliferation of pathogenic bacteria in humans depend on their capacity to impair host responses and acquire nutrients in a hostile environment. Among such nutrients is heme, a co-factor for oxygen storage, electron transport, photosynthesis, and redox biochemistry, which is indispensable for life. Porphyromonas gingivalis is the major human bacterial pathogen responsible for severe periodontitis. It recruits heme through HmuY, which sequesters heme from host carriers and delivers it to its cognate outer-membrane transporter, the TonB-dependent receptor HmuR. Here we report that heme binding does not significantly affect the secondary structure of HmuY. The crystal structure of heme-bound HmuY reveals a new all-beta fold mimicking a right hand. The thumb and fingers pinch heme iron through two apical histidine residues, giving rise to highly symmetric octahedral iron co-ordination. The tetrameric quaternary arrangement of the protein found in the crystal structure is consistent with experiments in solution. It shows that thumbs and fingertips, and, by extension, the bound heme groups, are shielded from competing heme-binding proteins from the host. This may also facilitate heme transport to HmuR for internalization. HmuY, both in its apo- and in its heme-bound forms, is resistant to proteolytic digestion by trypsin and the major secreted proteases of P. gingivalis, gingipains K and R. It is also stable against thermal and chemical denaturation. In conclusion, these studies reveal novel molecular properties of HmuY that are consistent with its role as a putative virulence factor during bacterial infection.

  18. Performance characteristics of anthropomorphic prosthetic hands.

    Science.gov (United States)

    Belter, Joseph T; Dollar, Aaron M

    2011-01-01

    In this paper we set forth a review of performance characteristics for both common commercial prosthetics as well as anthropomorphic research devices. Based on these specifications as well as surveyed results from prosthetic users, ranges of hand attributes are evaluated and discussed. End user information is used to describe the performance requirements for prosthetic hands for clinical use. © 2011 IEEE

  19. Prosthetic Hand Lifts Heavy Loads

    Science.gov (United States)

    Carden, James R.; Norton, William; Belcher, Jewell G.; Vest, Thomas W.

    1991-01-01

    Prosthetic hand designed to enable amputee to lift diverse heavy objects like rocks and logs. Has simple serrated end effector with no moving parts. Prosthesis held on forearm by system of flexible straps. Features include ruggedness, simplicity, and relatively low cost.

  20. Pre-prosthetic surgical alterations in maxillectomy to enhance the prosthetic prognoses as part of rehabilitation of oral cancer patient.

    Science.gov (United States)

    El Fattah, H; Zaghloul, A; Pedemonte, E; Escuin, T

    2012-03-01

    After maxillectomy, prosthetic restoration of the resulting defect is an essential step because it signals the beginning of patient's rehabilitation. The obturator used to restore the defect should be comfortable, restore adequate speech, deglutition, mastication, and be cosmetically acceptable, success will depend on the size and location of the defect and the quantity and integrity of the remaining structures, in addition to pre-prosthetic surgical preparation of defect site. Preoperative cooperation between the oncologist surgeon and the maxillofacial surgeon may allow obturation of a resultant defect by preservation of the premaxilla or the tuberosity on the defect side and maintaining the alveolar bone or teeth adjacent to the defect. This study evaluates the importance of pre-prosthetic surgical alterations at the time maxillectomy on the enhancement of the prosthetic prognoses as part of the rehabilitation of oral cancer patient. The study was carried out between 2003- 2008, on 66 cancer patients(41 male-25 female) age ranged from 33 to 72 years, at National Cancer Institute, Cairo University, whom underwent maxillectomy surgery to remove malignant tumor as a part of cancer treatment. Patients were divided in two groups. Group A: Resection of maxilla followed by preprosthetic surgical preparation. Twenty-four cancer patients (13 male - 11 female). Group B: Resection of maxilla without any preprosthetic surgical preparation. Forty-two cancer patients (28 male-14 female). Outcome variables measured included facial contour and aesthetic results, speech understandability, ability to eat solid foods, oronasal separation, socializing outside the home, and return-to-work status. Flap success and donor site morbidity were also studied. To improve the prosthetic restoration of maxillary defect resulting maxillary resection as part treatment of maxillofacial tumor depends on the close cooperation between prosthodontist and surgeon, by combination of pre-prosthetic

  1. The development of measurement tools for prosthetic eye research.

    Science.gov (United States)

    Pine, Keith Raymond; Sloan, Brian; Jacobs, Robert John

    2013-01-01

    The aim was to develop tools to measure the condition of ocular prostheses and the socket's response to prosthetic eyewear. A novel staining technique for displaying deposits on prosthetic eyes was developed. Equal interval perceptual grading scales for measuring inferior palpebral conjunctival inflammation, and anterior and posterior stained surface deposits on prosthetic eyes were developed from 800 photographs of 43 volunteers. The photographs for each scale were chosen by the authors. A group of four ophthalmologists, three optometrists and three senior students was consulted about selection criteria and asked to position the photographs along a 1.5 m rule to determine equal intervals. Photographs judged not to represent exactly equal perceptual intervals were exchanged with others from the original pool. The final scales (a five-photograph scale for inflammation and two 11 photograph scales for deposits) were assessed for inter-rater reliability and test-retest reliability by groups of senior optometry students. Standard deviations for inter-rater reliability tests were 0.52 scale units for the inflammation scale, 0.99 for the anterior surface deposits scale and 1.03 for the posterior surface deposits scale. The standard deviation of the test-retest differences for inflammation was 0.6 scale units and for both anterior and posterior surface deposits it was 0.71. A novel technique for displaying and measuring the intensity and extent of deposit formation on prosthetic eye surfaces has been described. The two equal interval perceptual grading scales that have been developed to quantify the extent of deposit formation together with the equal interval perceptual scale for grading severity of palpebral conjunctival inflammation will for the first time allow the effects of prosthetic eye wear to be evaluated. Further research to validate the scale for palpebral conjunctival inflammation in a clinical setting is recommended. The technique for staining deposits on

  2. Resonance Raman studies of the heme active site of the homodimeric myoglobin from Nassa mutabilis: a peculiar case.

    Science.gov (United States)

    Smulevich, G; Mantini, A R; Paoli, M; Coletta, M; Geraci, G

    1995-06-06

    A spectroscopic investigation by resonance Raman has been carried out at pH 7.0 in 0.1 M phosphate buffer on the cooperative homodimeric myoglobin from Nassa mutabilis. The study has been performed on the unligated ferrous form, as well as on the ligated species MbO2 and MbC, and on the ferric form met-Mb. Two v(C = C) vinyl stretching modes have been observed in all the investigated forms, reflecting different degrees of vinyl conjugation with the porphyrin ring, as a consequence of a strongly asymmetric environment for the two side groups of the heme. Furthermore, the ferric form displays a hexacoordinate low-spin heme, which suggests the presence of an endogenous ligand bound to the Fe atom. The frequency of the v(Fe-Im) stretching mode of Mb from Nassa mutabilis shifts down by 4 cm-1 as compared with that of horse heart myoglobin, reflecting a protein-induced proximal strain as a result of heme-heme interaction due to the close proximity of the two hemes in the dimer. The lower frequency of the v(Fe-Im) stretching mode agrees well with the lower affinity for oxygen binding found for Nassa mutabilis Mb and with the slight heme core expansion with respect to horse heart Mb, suggesting a critical role for the Fe-His bond on the heme's function and structure.

  3. Characterization of Heme Proteins Involved in Microbial Exoelectric Activity and Small Molecule-Sensing

    KAUST Repository

    Vogler, Malvina M.

    2018-01-01

    Heme proteins, also termed cytochromes, are a widespread class of metalloproteins containing an Fe-protoporphyrin IX cofactor. They perform numerous functions in nature such as oxygen-transport by hemoglobin, monooxygenation reactions catalyzed by Cytochrome P-450, and electron transfer reactions during photosynthesis. The differences between proteincofactor binding characteristics and the cofactor environment greatly influence the extensive range of functions. In this dissertation, proteins from the Mtr pathway of Shewanella oneidensis are characterized. These c-type cytochromes contain multiple heme cofactors per protein molecule that covalently attach to the protein amino acid sequence and are involved in electron transfer to extracellular metal oxides during anaerobic conditions. Successful recombinant expression of pathway components MtrC and MtrA is achieved in Escherichia coli. Heme-dependent gel staining and UV/Vis spectroscopy show characteristic c-type cytochrome characteristics. Mass spectrometry confirms that the correct extensive post-translational modifications were performed and the ten heme groups were incorporated per protein of MtrC and MtrA and the correct lipid-anchor was attached to extracellular MtrC. Raman spectroscopy measurements of MtrA provide intriguing structural information and highlight the strong influence of the heme cofactors within the protein structure. Next, an Arabidopsis thaliana protein is analyzed. It was previously identified via a motif search of the plant genome, based on conserved residues in the H4 NOX pocket. Here, the incorporation of a heme b cofactor is confirmed. UV/Vis spectroscopy under anaerobic conditions demonstrates reversible binding of nitric oxide to the heme iron and depicts the previously published characteristic absorption maxima for other H-NOX proteins.

  4. Investigations of ultrafast ligand rebinding to heme and heme proteins using temperature and strong magnetic field perturbations

    Science.gov (United States)

    Zhang, Zhenyu

    This thesis is written to summarize investigations of the mechanisms that underlie the kinetics of diatomic ligand rebinding to the iron atom of the heme group, which is chelated inside heme proteins. The family of heme proteins is a major object of studies for several branches of scientific research activity. Understanding the ligand binding mechanisms and pathways is one of the major goals for biophysics. My interests mainly focus on the physics of this ligand binding process. Therefore, to investigate the problem, isolated from the influence of the protein matrix, Fe-protophorphyrin IX is chosen as the prototype system in my studies. Myoglobin, the most extensively and intensively studied protein, is another ideal system that allows coupling the protein polypeptide matrix into the investigation. A technique to synchro-lock two laser pulse trains electronically is applied to our pump-probe spectroscopic studies. Based on this technique, a two color, fs/ps pump-probe system is developed which extends the temporal window for our investigation to 13ns and fills a gap existing in previous pump-probe investigations. In order to apply this newly-developed pump-probe laser system to implement systematic studies on the kinetics of diatomic ligand (NO, CO, O2) rebinding to heme and heme proteins, several experimental setups are utilized. In Chapter 1, the essential background knowledge, which helps to understand the iron-ligand interaction, is briefly described. In Chapter 2, in addition to a description of the preparation protocols of protein samples and details of the method for data analysis, three home-made setups are described, which include: a picosecond laser regenerative amplifier, a pump-probe application along the bore (2-inch in diameter) of a superconducting magnet and a temperature-controllable cryostat for spinning sample cell. Chapter 3 presents high magnetic field studies of several heme-ligand or protein-ligand systems. Pump-probe spectroscopy is used to

  5. Modeling and computations of the intramolecular electron transfer process in the two-heme protein cytochrome c4

    DEFF Research Database (Denmark)

    Natzmutdinov, Renat R.; Bronshtein, Michael D.; Zinkicheva, Tamara T.

    2012-01-01

    performed computational modeling of the intramolecular ET process by a combination of density functional theory (DFT) and quantum mechanical charge transfer theory to disclose reasons for this difference. We first address the electronic structures of the model heme core with histidine and methionine axial...... force were determined using dielectric continuum models. We then calculated the electronic transmission coefficient of the intramolecular ET rate using perturbation theory combined with the electronic wave functions determined by the DFT calculations for different heme group orientations and Fe......The di-heme protein Pseudomonas stutzeri cytochrome c4 (cyt c4) has emerged as a useful model for studying long-range protein electron transfer (ET). Recent experimental observations have shown a dramatically different pattern of intramolecular ET between the two heme groups in different local...

  6. Uncemented allograft-prosthetic composite reconstruction of the proximal femur

    Directory of Open Access Journals (Sweden)

    Li Min

    2014-01-01

    Full Text Available Background: Allograft-prosthetic composite can be divided into three groups names cemented, uncemented, and partially cemented. Previous studies have mainly reported outcomes in cemented and partially cemented allograft-prosthetic composites, but have rarely focused on the uncemented allograft-prosthetic composites. The objectives of our study were to describe a surgical technique for using proximal femoral uncemented allograft-prosthetic composite and to present the radiographic and clinical results. Materials and Methods: Twelve patients who underwent uncemented allograft-prosthetic composite reconstruction of the proximal femur after bone tumor resection were retrospectively evaluated at an average followup of 24.0 months. Clinical records and radiographs were evaluated. Results: In our series, union occurred in all the patients (100%; range 5-9 months. Until the most recent followup, there were no cases with infection, nonunion of the greater trochanter, junctional bone resorption, dislocation, allergic reaction, wear of acetabulum socket, recurrence, and metastasis. But there were three periprosthetic fractures which were fixed using cerclage wire during surgery. Five cases had bone resorption in and around the greater trochanter. The average Musculoskeletal Tumor Society (MSTS score and Harris hip score (HHS were 26.2 points (range 24-29 points and 80.6 points (range 66.2-92.7 points, respectively. Conclusions: These results showed that uncemented allograft-prosthetic composite could promote bone union through compression at the host-allograft junction and is a good choice for proximal femoral resection. Although this technology has its own merits, long term outcomes are yet not validated.

  7. Kidney injury and heme oxygenase-1

    Directory of Open Access Journals (Sweden)

    Hai-xing MAI

    2012-02-01

    Full Text Available     Heme oxygenase-1 (HO-1 is one of the main pathways to degrade heme in mammals, and the main degradation products are free iron (Fe2+, carbon monoxide (CO, and bilirubin. Heme plays an important role in promoting cell survival, circulation of intracellular substrates, and immune regulation. Previous studies suggest that HO-1 pathway is an important internal factor in determining the susceptibility and severity of acute kidney injury (AKI. The induction of HO-1 expression can attenuate the severity of renal ischemia-reperfusion injury (IRI, and the inhibition of HO-1 expression will aggravate IRI. The present article summarizes the latest advances in research abroad and at home on protective mechanism by which HO-1 prevents AKI to further deepen our understanding of the role of HO-1 in the treatment of AKI.   

  8. Structures of the Substrate-free and Product-bound Forms of HmuO, a Heme Oxygenase from Corynebacterium diphtheriae

    Science.gov (United States)

    Unno, Masaki; Ardèvol, Albert; Rovira, Carme; Ikeda-Saito, Masao

    2013-01-01

    Heme oxygenase catalyzes the degradation of heme to biliverdin, iron, and carbon monoxide. Here, we present crystal structures of the substrate-free, Fe3+-biliverdin-bound, and biliverdin-bound forms of HmuO, a heme oxygenase from Corynebacterium diphtheriae, refined to 1.80, 1.90, and 1.85 Å resolution, respectively. In the substrate-free structure, the proximal and distal helices, which tightly bracket the substrate heme in the substrate-bound heme complex, move apart, and the proximal helix is partially unwound. These features are supported by the molecular dynamic simulations. The structure implies that the heme binding fixes the enzyme active site structure, including the water hydrogen bond network critical for heme degradation. The biliverdin groups assume the helical conformation and are located in the heme pocket in the crystal structures of the Fe3+-biliverdin-bound and the biliverdin-bound HmuO, prepared by in situ heme oxygenase reaction from the heme complex crystals. The proximal His serves as the Fe3+-biliverdin axial ligand in the former complex and forms a hydrogen bond through a bridging water molecule with the biliverdin pyrrole nitrogen atoms in the latter complex. In both structures, salt bridges between one of the biliverdin propionate groups and the Arg and Lys residues further stabilize biliverdin at the HmuO heme pocket. Additionally, the crystal structure of a mixture of two intermediates between the Fe3+-biliverdin and biliverdin complexes has been determined at 1.70 Å resolution, implying a possible route for iron exit. PMID:24106279

  9. Prosthetics & Orthotics Manufacturing Initiative (POMI)

    Science.gov (United States)

    2012-12-21

    have promise in allowing sockets to be adjusted after manufacture. The most likely configuration involves placing shape-memory foams , which are...demonstrated that the liquid carbon dioxide-based system could provide cooling through the thermal resistances of the viscoelastic liner now commonly worn...When a patient’s residual limb is covered by the prosthetic system, including a viscoelastic liner, the limb is robbed of all natural mechanisms

  10. Bar-holding prosthetic limb

    Science.gov (United States)

    Vest, Thomas W. (Inventor); Norton, William E. (Inventor); Belcher, Jewell G. (Inventor); Carden, James R. (Inventor)

    1992-01-01

    A prosthetic device for below-the-elbow amputees is disclosed. The device has a removable effector, which is attached to the end of an arm cuff. The effector is comprised of a pair of C-shaped members that are oriented so as to face each other. Working in concert, the C-shaped members are able to hold a bar such as a chainsaw handle. A flat spring is fitted around the C-shaped members to hold them together.

  11. Molecular Simulations of Porphyrins and Heme Proteins

    Energy Technology Data Exchange (ETDEWEB)

    SHELNUTT,JOHN A.

    2000-01-18

    An overview of the use of classical mechanical molecular simulations of porphyrins, hydroporphyrins, and heme proteins is given. The topics cover molecular mechanics calculations of structures and conformer energies of porphyrins, energies of barriers for interconversion between stable conformers, molecular dynamics of porphyrins and heme proteins, and normal-coordinate structural analysis of experimental and calculated porphyrin structures. Molecular mechanics and dynamics are currently a fertile area of research on porphyrins. In the future, other computational methods such as Monte Carlo simulations, which have yet to be applied to porphyrins, will come into use and open new avenues of research into molecular simulations of porphyrins.

  12. Identification of the receptor scavenging hemopexin-heme complexes

    DEFF Research Database (Denmark)

    Hvidberg, Vibeke; Maniecki, Maciej B; Jacobsen, Christian

    2005-01-01

    Heme released from heme-binding proteins on internal hemorrhage, hemolysis, myolysis, or other cell damage is highly toxic due to oxidative and proinflammatory effects. Complex formation with hemopexin, the high-affinity heme-binding protein in plasma and cerebrospinal fluid, dampens these effects...

  13. Red meat and colon cancer : how dietary heme initiates hyperproliferation

    NARCIS (Netherlands)

    IJssennagger, N.

    2012-01-01

    Colorectal cancer is a leading cause of cancer deaths in Western countries. The risk to develop colorectal cancer is associated with the intake of red meat. Red meat contains the porphyrin pigment heme. Heme is an irritant for the colonic wall and it is previously shown that the addition of heme to

  14. The Quantum Mixed-Spin Heme State of Barley Peroxidase: A Paradigm for Class III Peroxidases

    Energy Technology Data Exchange (ETDEWEB)

    Howes, B.D.; Ma, J.; Marzocchi, M.P.; Schiodt, C.B.; Shelnutt, J.A.; Smulevich, G.; Welinder, K.G.; Zhang, J.

    1999-03-23

    Electronic absorption and resonance Raman (RR) spectra of the ferric form of barley grain peroxidase (BP 1) at various pH values both at room temperature and 20 K are . reported, together with EPR spectra at 10 K. The ferrous forms and the ferric complex with fluoride have also been studied. A quantum mechanically mixed-spin (QS) state has been identified. The QS heme species co-exists with 6- and 5-cHS heroes; the relative populations of these three spin states are found to be dependent on pH and temperature. However, the QS species remains in all cases the dominant heme spin species. Barley peroxidase appears to be further characterized by a splitting of the two vinyl stretching modes, indicating that the vinyl groups are differently conjugated with the porphyrin. An analysis of the presently available spectroscopic data for proteins from all three peroxidase classes suggests that the simultaneous occurrence of the QS heme state as well as the splitting of the two vinyl stretching modes is confined to class III enzymes. The former point is discussed in terms of the possible influences of heme deformations on heme spin state. It is found that moderate saddling alone is probably not enough to cause the QS state, although some saddling maybe necessary for the QS state.

  15. Detection and identification of heme c-modified peptides by histidine affinity chromatography, high-performance liquid chromatography-mass spectrometry, and database searching.

    Science.gov (United States)

    Merkley, Eric D; Anderson, Brian J; Park, Jea; Belchik, Sara M; Shi, Liang; Monroe, Matthew E; Smith, Richard D; Lipton, Mary S

    2012-12-07

    Multiheme c-type cytochromes (proteins with covalently attached heme c moieties) play important roles in extracellular metal respiration in dissimilatory metal-reducing bacteria. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of c-type cytochromes is hindered by the presence of multiple heme groups, since the heme c modified peptides are typically not observed or, if observed, not identified. Using a recently reported histidine affinity chromatography (HAC) procedure, we enriched heme c tryptic peptides from purified bovine heart cytochrome c, two bacterial decaheme cytochromes, and subjected these samples to LC-MS/MS analysis. Enriched bovine cytochrome c samples yielded 3- to 6-fold more confident peptide-spectrum matches to heme c containing peptides than unenriched digests. In unenriched digests of the decaheme cytochrome MtoA from Sideroxydans lithotrophicus ES-1, heme c peptides for 4 of the 10 expected sites were observed by LC-MS/MS; following HAC fractionation, peptides covering 9 out of 10 sites were obtained. Heme c peptide spiked into E. coli lysates at mass ratios as low as 1×10(-4) was detected with good signal-to-noise after HAC and LC-MS/MS analysis. In addition to HAC, we have developed a proteomics database search strategy that takes into account the unique physicochemical properties of heme c peptides. The results suggest that accounting for the double thioether link between heme c and peptide, and the use of the labile heme fragment as a reporter ion, can improve database searching results. The combination of affinity chromatography and heme-specific informatics yielded increases in the number of peptide-spectrum matches of 20-100-fold for bovine cytochrome c.

  16. Structure of the cytochrome complex SoxXA of Paracoccus pantotrophus, a heme enzyme initiating chemotrophic sulfur oxidation.

    Science.gov (United States)

    Dambe, Tresfore; Quentmeier, Armin; Rother, Dagmar; Friedrich, Cornelius; Scheidig, Axel J

    2005-12-01

    The sulfur-oxidizing enzyme system (Sox) of the chemotroph Paracoccus pantotrophus is composed of several proteins, which together oxidize hydrogen sulfide, sulfur, thiosulfate or sulfite and transfers the gained electrons to the respiratory chain. The hetero-dimeric cytochrome c complex SoxXA functions as heme enzyme and links covalently the sulfur substrate to the thiol of the cysteine-138 residue of the SoxY protein of the SoxYZ complex. Here, we report the crystal structure of the c-type cytochrome complex SoxXA. The structure could be solved by molecular replacement and refined to a resolution of 1.9A identifying the axial heme-iron coordination involving an unusual Cys-251 thiolate of heme2. Distance measurements between the three heme groups provide deeper insight into the electron transport inside SoxXA and merge in a better understanding of the initial step of the aerobic sulfur oxidation process in chemotrophic bacteria.

  17. Cognitive Control Signals for Neural Prosthetics

    National Research Council Canada - National Science Library

    S. Musallam; B. D. Corneil; B. Greger; H. Scherberger; R. A. Andersen

    2004-01-01

    Recent development of neural prosthetics for assisting paralyzed patients has focused on decoding intended hand trajectories from motor cortical neurons and using this signal to control external devices...

  18. Computer Aided Facial Prosthetics Manufacturing System

    Directory of Open Access Journals (Sweden)

    Peng H.K.

    2016-01-01

    Full Text Available Facial deformities can impose burden to the patient. There are many solutions for facial deformities such as plastic surgery and facial prosthetics. However, current fabrication method of facial prosthetics is high-cost and time consuming. This study aimed to identify a new method to construct a customized facial prosthetic. A 3D scanner, computer software and 3D printer were used in this study. Results showed that the new developed method can be used to produce a customized facial prosthetics. The advantages of the developed method over the conventional process are low cost, reduce waste of material and pollution in order to meet the green concept.

  19. Circuit For Control Of Electromechanical Prosthetic Hand

    Science.gov (United States)

    Bozeman, Richard J., Jr.

    1995-01-01

    Proposed circuit for control of electromechanical prosthetic hand derives electrical control signals from shoulder movements. Updated, electronic version of prosthesis, that includes two hooklike fingers actuated via cables from shoulder harness. Circuit built around favored shoulder harness, provides more dexterous movement, without incurring complexity of computer-controlled "bionic" or hydraulically actuated devices. Additional harness and potentiometer connected to similar control circuit mounted on other shoulder. Used to control stepping motor rotating hand about prosthetic wrist to one of number of angles consistent with number of digital outputs. Finger-control signals developed by circuit connected to first shoulder harness transmitted to prosthetic hand via sliprings at prosthetic wrist joint.

  20. Inhibition of Heme Peroxidases by Melamine

    Directory of Open Access Journals (Sweden)

    Pattaraporn Vanachayangkul

    2012-01-01

    Full Text Available In 2008 melamine-contaminated infant formula and dairy products in China led to over 50,000 hospitalizations of children due to renal injuries. In North America during 2007 and in Asia during 2004, melamine-contaminated pet food products resulted in numerous pet deaths due to renal failure. Animal studies have confirmed the potent renal toxicity of melamine combined with cyanuric acid. We showed previously that the solubility of melamine cyanurate is low at physiologic pH and ionic strength, provoking us to speculate how toxic levels of these compounds could be transported through the circulation without crystallizing until passing into the renal filtrate. We hypothesized that melamine might be sequestered by heme proteins, which could interfere with heme enzyme activity. Four heme peroxidase enzymes were selected for study: horseradish peroxidase (HRP, lactoperoxidase (LPO, and cyclooxygenase-1 and -2 (COX-1 and -2. Melamine exhibited noncompetitive inhibition of HRP (9.5±0.7mM, and LPO showed a mixed model of inhibition (14.5±4.7mM. The inhibition of HRP and LPO was confirmed using a chemiluminescent peroxidase assay. Melamine also exhibited COX-1 inhibition, but inhibition of COX-2 was not detected. Thus, our results demonstrate that melamine inhibits the activity of three heme peroxidases.

  1. Unsaturated glycerophospholipids mediate heme crystallization: biological implications for hemozoin formation in the kissing bug Rhodnius prolixus.

    Directory of Open Access Journals (Sweden)

    Renata Stiebler

    Full Text Available Hemozoin (Hz is a heme crystal produced by some blood-feeding organisms, as an efficient way to detoxify heme derived from hemoglobin digestion. In the triatomine insect Rhodnius prolixus, Hz is essentially produced by midgut extracellular phospholipid membranes known as perimicrovillar membranes (PMVM. Here, we investigated the role of commercial glycerophospholipids containing serine, choline and ethanolamine as headgroups and R. prolixus midgut lipids (RML in heme crystallization. All commercial unsaturated forms of phospholipids, as well as RML, mediated fast and efficient β-hematin formation by means of two kinetically distinct mechanisms: an early and fast component, followed by a late and slow one. The fastest reactions observed were induced by unsaturated forms of phosphatidylethanolamine (uPE and phosphatidylcholine (uPC, with half-lives of 0.04 and 0.7 minutes, respectively. β-hematin crystal morphologies were strikingly distinct among groups, with uPE producing homogeneous regular brick-shaped crystals. Interestingly, uPC-mediated reactions resulted in two morphologically distinct crystal populations: one less representative group of regular crystals, resembling those induced by uPE, and the other largely represented by crystals with numerous sharp edges and tapered ends. Heme crystallization reactions induced by RML were efficient, with a heme to β-hematin conversion rate higher than 70%, but clearly slower (t1/2 of 9.9-17.7 minutes than those induced by uPC and uPE. Interestingly, crystals produced by RML were homogeneous in shape and quite similar to those mediated by uPE. Thus, β-hematin formation can be rapidly and efficiently induced by unsaturated glycerophospholipids, particularly uPE and uPC, and may play a role on biological heme crystallization in R. prolixus midgut.

  2. Selective criteria for successful long-term prosthetic use.

    Science.gov (United States)

    Mueller, M J; Delitto, A

    1985-07-01

    The purpose of this study was to identify criteria contributing to successful long-term prosthetic use in patients with an amputation secondary to vascular disease. All elderly patients with a unilateral below-knee amputation or an above-knee amputation, secondary to vascular disease, seen in our clinic between 1977 and 1982 were included in this telephone survey. Of those contacted, 37 of 38 below-knee amputees (BKAs) and 7 of 18 above-knee amputees (AKAs) still wore their prostheses at least part of every day (success). We used a two-tailed chi-square to compare the success of the BKAs with the success of the AKAs. The BKAs were successful more often (X2 = 24.81, df = 1, p less than .001). All AKAs also were characterized according to age, time from prescription, obesity, ambulatory status, strength, range of motion, sex, general compliance, and medical problems after prosthetic prescription. Of these criteria, only compliance and medical problems after prescription showed a significant difference between successful and nonsuccessful long-term AKA prosthetic users (X2 = 5.76, df = 1, p less than .05 for each criterion). As the demands of quality assurance and diagnostic related groupings increase, these results can assist the physical therapy clinician in setting realistic goals for the geriatric amputee and help predict if the patient will be a successful prosthetic user.

  3. Multicopper manganese oxidase accessory proteins bind Cu and heme.

    Science.gov (United States)

    Butterfield, Cristina N; Tao, Lizhi; Chacón, Kelly N; Spiro, Thomas G; Blackburn, Ninian J; Casey, William H; Britt, R David; Tebo, Bradley M

    2015-12-01

    Multicopper oxidases (MCOs) catalyze the oxidation of a diverse group of metal ions and organic substrates by successive single-electron transfers to O2 via four bound Cu ions. MnxG, which catalyzes MnO2 mineralization by oxidizing both Mn(II) and Mn(III), is unique among multicopper oxidases in that it carries out two energetically distinct electron transfers and is tightly bound to accessory proteins. There are two of these, MnxE and MnxF, both approximately 12kDa. Although their sequences are similar to those found in the genomes of several Mn-oxidizing Bacillus species, they are dissimilar to those of proteins with known function. Here, MnxE and MnxF are co-expressed independent of MnxG and are found to oligomerize into a higher order stoichiometry, likely a hexamer. They bind copper and heme, which have been characterized by electron paramagnetic resonance (EPR), X-ray absorption spectroscopy (XAS), and UV-visible (UV-vis) spectrophotometry. Cu is found in two distinct type 2 (T2) copper centers, one of which appears to be novel; heme is bound as a low-spin species, implying coordination by two axial ligands. MnxE and MnxF do not oxidize Mn in the absence of MnxG and are the first accessory proteins to be required by an MCO. This may indicate that Cu and heme play roles in electron transfer and/or Cu trafficking. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Consumer Guide for Amputees: A Guide to Lower Limb Prosthetics

    Science.gov (United States)

    Consumer Guide for Amputees: A Guide to Lower Limb Prosthetics: Part I -- Prosthetic Design: Basic Concepts Volume 8 · ... wanted to have available a comprehensive explanation of limb prosthetics written in easily understood language for amputee consumers. ...

  5. Endogenous Estrogen-Mediated Heme Oxygenase Regulation in Experimental Menopause

    Directory of Open Access Journals (Sweden)

    Anikó Pósa

    2015-01-01

    Full Text Available Estrogen deficiency is one of the main causes of age-associated diseases in the cardiovascular system. Female Wistar rats were divided into four experimental groups: pharmacologically ovariectomized, surgically ovariectomized, and 24-month-old intact aging animals were compared with a control group. The activity and expression of heme oxygenases (HO in the cardiac left ventricle, the concentrations of cardiac interleukin-6 (IL-6 and tumor necrosis factor-α (TNF-α, the myeloperoxidase (MPO activity in the cardiac left ventricle, and the effects of heme oxygenase blockade (by 24-hour and 1-hour pretreatment with tin-protoporphyrin IX, SnPP on the epinephrine and phentolamine-induced electrocardiogram ST segment changes in vivo were investigated. The cardiac HO activity and the expression of HO-1 and HO-2 were significantly decreased in the aged rats and after ovariectomy. Estrogen depletion was accompanied by significant increases in the expression of IL-6 and TNF-α. The aged and ovariectomized animals exhibited a significantly elevated MPO activity and a significant ST segment depression. After pretreatment with SnPP augmented ST segment changes were determined. These findings demonstrate that the sensitivity to cardiac ischemia in estrogen depletion models is associated with suppression of the activity and expression of the HO system and increases in the secretion of proinflammatory cytokines and biomarkers.

  6. Endogenous Estrogen-Mediated Heme Oxygenase Regulation in Experimental Menopause

    Science.gov (United States)

    Pósa, Anikó; Veszelka, Médea; Berkó, Anikó Magyariné; Baráth, Zoltán; Ménesi, Rudolf; Pávó, Imre; László, Ferenc; Varga, Csaba

    2015-01-01

    Estrogen deficiency is one of the main causes of age-associated diseases in the cardiovascular system. Female Wistar rats were divided into four experimental groups: pharmacologically ovariectomized, surgically ovariectomized, and 24-month-old intact aging animals were compared with a control group. The activity and expression of heme oxygenases (HO) in the cardiac left ventricle, the concentrations of cardiac interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), the myeloperoxidase (MPO) activity in the cardiac left ventricle, and the effects of heme oxygenase blockade (by 24-hour and 1-hour pretreatment with tin-protoporphyrin IX, SnPP) on the epinephrine and phentolamine-induced electrocardiogram ST segment changes in vivo were investigated. The cardiac HO activity and the expression of HO-1 and HO-2 were significantly decreased in the aged rats and after ovariectomy. Estrogen depletion was accompanied by significant increases in the expression of IL-6 and TNF-α. The aged and ovariectomized animals exhibited a significantly elevated MPO activity and a significant ST segment depression. After pretreatment with SnPP augmented ST segment changes were determined. These findings demonstrate that the sensitivity to cardiac ischemia in estrogen depletion models is associated with suppression of the activity and expression of the HO system and increases in the secretion of proinflammatory cytokines and biomarkers. PMID:26064421

  7. Rotationally actuated prosthetic helping hand

    Science.gov (United States)

    Norton, William E. (Inventor); Belcher, Jewell G., Jr. (Inventor); Carden, James R. (Inventor); West, Thomas W. (Inventor)

    1991-01-01

    A prosthetic device has been developed for below-the-elbow amputees. The device consists of a cuff, a stem, a housing, two hook-like fingers, an elastic band for holding the fingers together, and a brace. The fingers are pivotally mounted on a housing that is secured to the amputee's upper arm with the brace. The stem, which also contains a cam, is rotationally mounted within the housing and is secured to the cuff, which fits over the amputee's stump. By rotating the cammed stem between the fingers with the lower arm, the amputee can open and close the fingers.

  8. Cochlear Implant Using Neural Prosthetics

    Science.gov (United States)

    Gupta, Shweta; Singh, Shashi kumar; Dubey, Pratik Kumar

    2012-10-01

    This research is based on neural prosthetic device. The oldest and most widely used of these electrical, and often computerized, devices is the cochlear implant, which has provided hearing to thousands of congenitally deaf people in this country. Recently, the use of the cochlear implant is expanding to the elderly, who frequently suffer major hearing loss. More cutting edge are artificial retinas, which are helping dozens of blind people see, and ìsmartî artificial arms and legs that amputees can maneuver by thoughts alone, and that feel more like real limbs.Research, which curiosity led to explore frog legs dancing during thunderstorms, a snail shapedorgan in the inner ear, and how various eye cells react to light, have fostered an understanding of how to ìtalkî to the nervous system. That understanding combined with the miniaturization of electronics and enhanced computer processing has enabled prosthetic devices that often can bridge the gap in nerve signaling that is caused by disease or injury.

  9. PLANNING OF IMPLANTO‐PROSTHETIC STRUCTURES IN TOTAL EDENTATION ACCORDING TO BIOMECHANICAL CRITERIA

    OpenAIRE

    Vasile, N.; V. IBRIC‐CIORANU

    2013-01-01

    Scope of the study: Analysis of the biomechanical aspects of the implanto‐prosthetic structures for planning the treatment in cases of total edentation.Materials and method: The study was performed in the Clinics of Prosthetics and Oral Implantology, Military Emergency Hospital of Sibiu, on a group of mandibularly totally edentated patients. Planning of the implanto‐pros‐ thetic treatment had in view the peculiarities of total man‐ dibular edentation and observance of the biomechanical princi...

  10. Undergraduate prosthetics and orthotics teaching methods: A baseline for international comparison.

    Science.gov (United States)

    Aminian, Gholamreza; O'Toole, John M; Mehraban, Afsoon Hassani

    2015-08-01

    Education of Prosthetics and Orthotics is a relatively recent professional program. While there has been some work on various teaching methods and strategies in international medical education, limited publication exists within prosthetics and orthotics. To identify the teaching and learning methods that are used in Bachelor-level prosthetics and orthotics programs that are given highest priority by expert prosthetics and orthotics instructors from regions enjoying a range of economic development. Mixed method. The study partly documented by this article utilized a mixed method approach (qualitative and quantitative methods) within which each phase provided data for other phases. It began with analysis of prosthetics and orthotics curricula documents, which was followed by a broad survey of instructors in this field and then a modified Delphi process. The expert instructors who participated in this study gave high priority to student-centered, small group methods that encourage critical thinking and may lead to lifelong learning. Instructors from more developed nations placed higher priority on student's independent acquisition of prosthetics and orthotics knowledge, particularly in clinical training. Application of student-centered approaches to prosthetics and orthotics programs may be preferred by many experts, but there appeared to be regional differences in the priority given to different teaching methods. The results of this study identify the methods of teaching that are preferred by expert prosthetics and orthotics instructors from a variety of regions. This treatment of current instructional techniques may inform instructor choice of teaching methods that impact the quality of education and improve the professional skills of students. © The International Society for Prosthetics and Orthotics 2014.

  11. Heme oxygenase-1-generated biliverdin ameliorates experimental murine colitis.

    Science.gov (United States)

    Berberat, Pascal O; A-Rahim, Yousif I; Yamashita, Kenichiro; Warny, Michel M; Csizmadia, Eva; Robson, Simon C; Bach, Fritz H

    2005-04-01

    Heme oxygenase-1 (HO-1) seems to have an important protective role in acute and chronic inflammation. The products of heme catalysis, biliverdin/bilirubin, carbon monoxide (CO), and iron (that induces apoferritin) mediate the beneficial effects of HO-1. Blockade of HO-1 activity results in exacerbation of experimental colitis. We tested whether HO-1 has protective effects in the development of colitis and determined that specific enzymatic products of HO-1 are responsible for these effects. Colitis was induced by oral administration of dextran sodium sulfate (5%) to C57BL/6 mice for 7 days. HO-1 was up-regulated by cobalt-protoporphyrin (5 mg/kg, intraperitoneally). Biliverdin, exogenous CO, or the iron chelator desferrioxamine was administered to other groups. Cobalt-protoporphyrin treatment resulted in significant up-regulation of HO-1 protein in mucosal and submucosal cells. Induction of HO-1 was associated with significantly less loss of body weight in mice with induced colitis (-12% versus -22% in the control animals, P biliverdin administration (50 micromol/kg, 3 times per day, intraperitoneally). We conclude that heightened HO-1 expression or administration of biliverdin ameliorates dextran sodium sulfate-induced experimental colitis. Novel therapeutic strategies based on HO-1 and/or biliverdin administration may have use in inflammatory bowel disease.

  12. Modulation of Na+/K+ ATPase Activity by Hydrogen Peroxide Generated through Heme in L. amazonensis.

    Directory of Open Access Journals (Sweden)

    Nathália Rocco-Machado

    Full Text Available Leishmania amazonensis is a protozoan parasite that occurs in many areas of Brazil and causes skin lesions. Using this parasite, our group showed the activation of Na+/K+ ATPase through a signaling cascade that involves the presence of heme and protein kinase C (PKC activity. Heme is an important biomolecule that has pro-oxidant activity and signaling capacity. Reactive oxygen species (ROS can act as second messengers, which are required in various signaling cascades. Our goal in this work is to investigate the role of hydrogen peroxide (H2O2 generated in the presence of heme in the Na+/K+ ATPase activity of L. amazonensis. Our results show that increasing concentrations of heme stimulates the production of H2O2 in a dose-dependent manner until a concentration of 2.5 μM heme. To confirm that the effect of heme on the Na+/K+ ATPase is through the generation of H2O2, we measured enzyme activity using increasing concentrations of H2O2 and, as expected, the activity increased in a dose-dependent manner until a concentration of 0.1 μM H2O2. To investigate the role of PKC in this signaling pathway, we observed the production of H2O2 in the presence of its activator phorbol 12-myristate 13-acetate (PMA and its inhibitor calphostin C. Both showed no effect on the generation of H2O2. Furthermore, we found that PKC activity is increased in the presence of H2O2, and that in the presence of calphostin C, H2O2 is unable to activate the Na+/K+ ATPase. 100 μM of Mito-TEMPO was capable of abolishing the stimulatory effect of heme on Na+/K+ ATPase activity, indicating that mitochondria might be the source of the hydrogen peroxide production induced by heme. The modulation of L. amazonensis Na+/K+ ATPase by H2O2 opens new possibilities for understanding the signaling pathways of this parasite.

  13. A comparison of vacuum and KBM prosthetic fitting for unilateral transtibial amputees using the Gait Profile Score.

    Science.gov (United States)

    Kuntze Ferreira, Alana Elisabeth; Neves, Eduardo Borba

    2015-02-01

    The purpose of this study was to compare gait deviations between Kondylen Bettung Münster (KBM) and vacuum prosthetic fitting using the Gait Profile Score (GPS), the Movement Analysis Profile (MAP) and temporal-spatial parameters. Seventeen transtibial amputees that received their prosthesis from the Brazilian governmental health system participated in this study. Twelve of them used KBM prosthetic fitting on their prosthesis and five used vacuum prosthetic fitting. Kinematic and temporal-spatial parameters data were captured by a six-camera Motion Analysis system (Santa Rosa, CA). The results showed that the vacuum group walked faster than the KBM group but the differences in temporal-spatial parameters between them were not significant. The GPS for the intact limb (IL) and the overall GPS differentiated between the groups of prosthetic fitting. Hip flexion/extension and knee flexion/extension were higher in KBM group than in the vacuum group, although only knee flexion/extension for the intact limb revealed significant difference between the groups. In KBM group, the major deviations were in hip flexion/extension for both limbs, knee flexion/extension for both limbs and ankle dorsi/plantar flexion for the prosthetic limb. The vacuum group showed deviations especially in ankle dorsi/plantar flexion for both limbs, knee flexion/extension for the prosthetic limb and hip rotation for the prosthetic limb. Besides, the vacuum group was more symmetrical than the KBM group. This study concluded that subjects who used vacuum prosthetic fitting presented smaller gait deviations and a more symmetrical gait than those who used KBM prosthetic fitting. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Brain responses to acupuncture stimulation in the prosthetic hand of an amputee patient.

    Science.gov (United States)

    Lee, In-Seon; Jung, Won-Mo; Lee, Ye-Seul; Wallraven, Christian; Chae, Younbyoung

    2015-10-01

    This report describes the brain responses to acupuncture in an upper limb amputee patient. A 62-year-old male had previously undergone a lower left arm amputation following an electrical accident. Using functional MRI, we investigated brain responses to acupuncture stimulation in the aforementioned amputee under three conditions: (a) intact hand, (b) prosthetic hand (used by the patient), and (c) fake fabric hand. The patient described greater de qi sensation when he received acupuncture stimulation in his prosthetic hand compared to a fake hand, with both stimulations performed in a similar manner. We found enhanced brain activation in the insula and sensorimotor cortex in response to acupuncture stimulation in the amputee's prosthetic hand, while there was only minimal activation in the visual cortex in response to acupuncture stimulation in a fake hand. The enhanced brain responses to acupuncture stimulation of the patient's prosthetic hand might be derived from cortical reorganisation, as he has been using his prosthetic hand for over 40 years. Our findings suggest the possible use of acupuncture stimulation in a prosthetic hand as an enhanced sensory feedback mechanism, which may represent a new treatment approach for phantom limb pain. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  15. Role of Heme and Heme-Proteins in Trypanosomatid Essential Metabolic Pathways

    Directory of Open Access Journals (Sweden)

    Karina E. J. Tripodi

    2011-01-01

    Full Text Available Around the world, trypanosomatids are known for being etiological agents of several highly disabling and often fatal diseases like Chagas disease (Trypanosoma cruzi, leishmaniasis (Leishmania spp., and African trypanosomiasis (Trypanosoma brucei. Throughout their life cycle, they must cope with diverse environmental conditions, and the mechanisms involved in these processes are crucial for their survival. In this review, we describe the role of heme in several essential metabolic pathways of these protozoans. Notwithstanding trypanosomatids lack of the complete heme biosynthetic pathway, we focus our discussion in the metabolic role played for important heme-proteins, like cytochromes. Although several genes for different types of cytochromes, involved in mitochondrial respiration, polyunsaturated fatty acid metabolism, and sterol biosynthesis, are annotated at the Tritryp Genome Project, the encoded proteins have not yet been deeply studied. We pointed our attention into relevant aspects of these protein functions that are amenable to be considered for rational design of trypanocidal agents.

  16. Mimicking Heme Enzymes in the Solid State: Metal-Organic Materials with Selectively Encapsulated Heme

    Energy Technology Data Exchange (ETDEWEB)

    Larsen, Randy W; Wojtas, Lukasz; Perman, Jason; Musselman, Ronald L; Zaworotko, Michael J; Vetromile, Carissa M [USF

    2011-06-13

    To carry out essential life processes, nature has had to evolve heme enzymes capable of synthesizing and manipulating complex molecules. These proteins perform a plethora of chemical reactions utilizing a single iron porphyrin active site embedded within an evolutionarily designed protein pocket. We herein report the first class of metal–organic materials (MOMs) that mimic heme enzymes in terms of both structure and reactivity. The MOMzyme-1 class is based upon a prototypal MOM, HKUST-1, into which catalytically active metalloporphyrins are selectively encapsulated in a “ship-in-a-bottle” fashion within one of the three nanoscale cages that exist in HKUST-1. MOMs offer unparalleled levels of permanent porosity and their modular nature affords enormous diversity of structures and properties. The MOMzyme-1 class could therefore represent a new paradigm for heme biomimetic catalysis since it combines the activity of a homogeneous catalyst with the stability and recyclability of heterogeneous catalytic systems within a single material.

  17. Mimicking heme enzymes in the solid state: metal-organic materials with selectively encapsulated heme.

    Science.gov (United States)

    Larsen, Randy W; Wojtas, Lukasz; Perman, Jason; Musselman, Ronald L; Zaworotko, Michael J; Vetromile, Carissa M

    2011-07-13

    To carry out essential life processes, nature has had to evolve heme enzymes capable of synthesizing and manipulating complex molecules. These proteins perform a plethora of chemical reactions utilizing a single iron porphyrin active site embedded within an evolutionarily designed protein pocket. We herein report the first class of metal-organic materials (MOMs) that mimic heme enzymes in terms of both structure and reactivity. The MOMzyme-1 class is based upon a prototypal MOM, HKUST-1, into which catalytically active metalloporphyrins are selectively encapsulated in a "ship-in-a-bottle" fashion within one of the three nanoscale cages that exist in HKUST-1. MOMs offer unparalleled levels of permanent porosity and their modular nature affords enormous diversity of structures and properties. The MOMzyme-1 class could therefore represent a new paradigm for heme biomimetic catalysis since it combines the activity of a homogeneous catalyst with the stability and recyclability of heterogeneous catalytic systems within a single material.

  18. Holo- And Apo- Structures of Bacterial Periplasmic Heme Binding Proteins

    Energy Technology Data Exchange (ETDEWEB)

    Ho, W.W.; Li, H.; Eakanunkul, S.; Tong, Y.; Wilks, A.; Guo, M.; Poulos, T.L.

    2009-06-01

    An essential component of heme transport in Gram-negative bacterial pathogens is the periplasmic protein that shuttles heme between outer and inner membranes. We have solved the first crystal structures of two such proteins, ShuT from Shigella dysenteriae and PhuT from Pseudomonas aeruginosa. Both share a common architecture typical of Class III periplasmic binding proteins. The heme binds in a narrow cleft between the N- and C-terminal binding domains and is coordinated by a Tyr residue. A comparison of the heme-free (apo) and -bound (holo) structures indicates little change in structure other than minor alterations in the heme pocket and movement of the Tyr heme ligand from an 'in' position where it can coordinate the heme iron to an 'out' orientation where it points away from the heme pocket. The detailed architecture of the heme pocket is quite different in ShuT and PhuT. Although Arg{sup 228} in PhuT H-bonds with a heme propionate, in ShuT a peptide loop partially takes up the space occupied by Arg{sup 228}, and there is no Lys or Arg H-bonding with the heme propionates. A comparison of PhuT/ShuT with the vitamin B{sub 12}-binding protein BtuF and the hydroxamic-type siderophore-binding protein FhuD, the only two other structurally characterized Class III periplasmic binding proteins, demonstrates that PhuT/ShuT more closely resembles BtuF, which reflects the closer similarity in ligands, heme and B{sub 12}, compared with ligands for FhuD, a peptide siderophore.

  19. 21 CFR 890.3420 - External limb prosthetic component.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false External limb prosthetic component. 890.3420... External limb prosthetic component. (a) Identification. An external limb prosthetic component is a device... total prosthesis. Examples of external limb prosthetic components include the following: Ankle, foot...

  20. Development of prosthetic arm with pneumatic prosthetic hand and tendon-driven wrist.

    Science.gov (United States)

    Takeda, Hiroyuki; Tsujiuchi, Nobutaka; Koizumi, Takayuki; Kan, Hiroto; Hirano, Masanori; Nakamura, Yoichiro

    2009-01-01

    Recently, various prosthetic arms have been developed, but few are both attractive and functional. Considering human coexistence, prosthetic arms must be both safe and flexible. In this research, we developed a novel prosthetic arm with a five-fingered prosthetic hand using our original pneumatic actuators and a slender tendon-driven wrist using a wire drive and two small motors. Because the prosthetic hand's driving source is comprised of small pneumatic actuators, the prosthetic hand is safe when it makes contact with people; it can also operate flexibly. In addition, the arm has a tendon-driven wrist to expand its motion space and to perform many operations. First, we explain the pneumatic hand's drive mechanism and its tendon-driven wrist. Next, we identify the characteristics of the hand and the wrist and construct a control system for this arm and verify its control performance.

  1. Structural characterization of human heme oxygenase-1 in complex with azole-based inhibitors.

    Science.gov (United States)

    Rahman, Mona N; Vlahakis, Jason Z; Roman, Gheorghe; Vukomanovic, Dragic; Szarek, Walter A; Nakatsu, Kanji; Jia, Zongchao

    2010-03-01

    The development of inhibitors specific for heme oxygenases (HO) aims to provide powerful tools in understanding the HO system. Based on the lead structure (2S, 4S)-2-[2-(4-chlorophenyl)ethyl]-2-[(1H-imidazol-1-yl)methyl]-4-[((4-aminophenyl)thio)methyl]-1,3-dioxolane (azalanstat, QC-1) we have synthesized structural modifications to develop novel and selective HO inhibitors. The structural study of human HO-1 (hHO-1) in complex with a select group of the inhibitors was initiated using X-ray crystallographic techniques. Comparison of the structures of four such compounds each in complex with hHO-1 revealed a common binding mode, despite having different structural fragments. The compounds bind to the distal side of heme through an azole "anchor" which coordinates with the heme iron. An expansion of the distal pocket, mainly due to distal helix flexibility, allows accommodation of the compounds without displacing heme or the critical Asp140 residue. Rather, binding displaces a catalytically critical water molecule and disrupts an ordered hydrogen-bond network involving Asp140. The presence of a triazole "anchor" may provide further stability via a hydrogen bond with the protein. A hydrophobic pocket acts to stabilize the region occupied by the phenyl or adamantanyl moieties of these compounds. Further, a secondary hydrophobic pocket is formed via "induced fit" to accommodate bulky substituents at the 4-position of the dioxolane ring. Copyright 2009 Elsevier Inc. All rights reserved.

  2. Protein oxidation mediated by heme-induced active site conversion specific for heme-regulated transcription factor, iron response regulator.

    Science.gov (United States)

    Kitatsuji, Chihiro; Izumi, Kozue; Nambu, Shusuke; Kurogochi, Masaki; Uchida, Takeshi; Nishimura, Shin-ichiro; Iwai, Kazuhiro; O'Brian, Mark R; Ikeda-Saito, Masao; Ishimori, Koichiro

    2016-01-05

    The Bradyrhizobium japonicum transcriptional regulator Irr (iron response regulator) is a key regulator of the iron homeostasis, which is degraded in response to heme binding via a mechanism that involves oxidative modification of the protein. Here, we show that heme-bound Irr activates O2 to form highly reactive oxygen species (ROS) with the "active site conversion" from heme iron to non-heme iron to degrade itself. In the presence of heme and reductant, the ROS scavenging experiments show that Irr generates H2O2 from O2 as found for other hemoproteins, but H2O2 is less effective in oxidizing the peptide, and further activation of H2O2 is suggested. Interestingly, we find a time-dependent decrease of the intensity of the Soret band and appearance of the characteristic EPR signal at g = 4.3 during the oxidation, showing the heme degradation and the successive formation of a non-heme iron site. Together with the mutational studies, we here propose a novel "two-step self-oxidative modification" mechanism, during which O2 is activated to form H2O2 at the heme regulatory motif (HRM) site and the generated H2O2 is further converted into more reactive species such as ·OH at the non-heme iron site in the His-cluster region formed by the active site conversion.

  3. Advanced upper limb prosthetic devices: implications for upper limb prosthetic rehabilitation.

    Science.gov (United States)

    Resnik, Linda; Meucci, Marissa R; Lieberman-Klinger, Shana; Fantini, Christopher; Kelty, Debra L; Disla, Roxanne; Sasson, Nicole

    2012-04-01

    The number of catastrophic injuries caused by improvised explosive devices in the Afghanistan and Iraq Wars has increased public, legislative, and research attention to upper limb amputation. The Department of Veterans Affairs (VA) has partnered with the Defense Advanced Research Projects Agency and DEKA Integrated Solutions to optimize the function of an advanced prosthetic arm system that will enable greater independence and function. In this special communication, we examine current practices in prosthetic rehabilitation including trends in adoption and use of prosthetic devices, financial considerations, and the role of rehabilitation team members in light of our experiences with a prototype advanced upper limb prosthesis during a VA study to optimize the device. We discuss key challenges in the adoption of advanced prosthetic technology and make recommendations for service provision and use of advanced upper limb prosthetics. Rates of prosthetic rejection are high among upper limb amputees. However, these rates may be reduced with sufficient training by a highly specialized, multidisciplinary team of clinicians, and a focus on patient education and empowerment throughout the rehabilitation process. There are significant challenges emerging that are unique to implementing the use of advanced upper limb prosthetic technology, and a lack of evidence to establish clinical guidelines regarding prosthetic prescription and treatment. Finally, we make recommendations for future research to aid in the identification of best practices and development of policy decisions regarding insurance coverage of prosthetic rehabilitation. Copyright © 2012 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.

  4. Relationship between natural and heme-mediated antibody polyreactivity

    Energy Technology Data Exchange (ETDEWEB)

    Hadzhieva, Maya; Vassilev, Tchavdar [Stephan Angelov Institute of Microbiology, Bulgarian Academy of Sciences, Sofia 1113 (Bulgaria); Bayry, Jagadeesh; Kaveri, Srinivas; Lacroix-Desmazes, Sébastien [Sorbonne Universités, UPMC Univ Paris 06, UMR-S 1138, Centre de Recherche des Cordeliers, F-75006 Paris (France); INSERM, UMR-S 1138, F-75006 Paris (France); Université Paris Descartes, Sorbonne Paris Cité, UMR-S 1138, F-75006 Paris (France); Dimitrov, Jordan D., E-mail: jordan.dimitrov@crc.jussieu.fr [Sorbonne Universités, UPMC Univ Paris 06, UMR-S 1138, Centre de Recherche des Cordeliers, F-75006 Paris (France); INSERM, UMR-S 1138, F-75006 Paris (France); Université Paris Descartes, Sorbonne Paris Cité, UMR-S 1138, F-75006 Paris (France)

    2016-03-25

    Polyreactive antibodies represent a considerable fraction of the immune repertoires. Some antibodies acquire polyreactivity post-translationally after interaction with various redox-active substances, including heme. Recently we have demonstrated that heme binding to a naturally polyreactive antibody (SPE7) results in a considerable broadening of the repertoire of recognized antigens. A question remains whether the presence of certain level of natural polyreactivity of antibodies is a prerequisite for heme-induced further extension of antigen binding potential. Here we used a second monoclonal antibody (Hg32) with unknown specificity and absence of intrinsic polyreactivity as a model to study the potential of heme to induce polyreactivity of antibodies. We demonstrated that exposure to heme greatly extends the antigen binding potential of Hg32, suggesting that the intrinsic binding promiscuity is not a prerequisite for the induction of polyreactivity by heme. In addition we compared the kinetics and thermodynamics of the interaction of heme-exposed antibodies with a panel of unrelated antigens. These analyses revealed that the two heme-sensitive antibodies adopt different mechanisms of binding to the same set of antigens. This study contributes to understanding the phenomenon of induced antibody polyreactivity. The data may also be of importance for understanding of physiological and pathological roles of polyreactive antibodies. - Highlights: • Exposure of certain monoclonal IgE antibodies to heme results in gain of antigen binding polyreactivity. • Natural polyreactivity of antibodies is dispensable for acquisition of polyreactivity through interaction with heme. • Heme-induced monoclonal IgE antibodies differ in their thermodynamic mechanisms of antigen recognition.

  5. Lymphatic opacification in the prosthetic hip

    Energy Technology Data Exchange (ETDEWEB)

    Bloom, R.A.; Gheorghiu, D. (Hadassah Univ. Hospital, Jerusalem (Israel). Dept. of Radiology); Krausz, Y. (Hadassah Univ. Hospital, Jerusalem (Israel). Dept. of Nuclear Medicine)

    1991-01-01

    A retrospective analysis of 52 patients with hip pain following total hip replacement was made. Each of them was evaluated by plain radiographs, technetium 99m pyrophosphate scans, arthrography with plain film substraction technique, and culture of joint fluid. In 30 cases there was evidence of prosthetic loosening, and in 21 of these lymphangeal opacification during arthrography was seen. In 15 cases with lymphongeal opacification the daignosis of prosthetic loosening was subsequently confirmed by prosthetic revision. In none of the 22 cases in which no evidence of prosthetic loosening was seen was there lymphatic opacification. It is concluded that lymphatic opacification during arthrography for pain following total hip prosthesis is a valuable ancillary sign of loosening. (orig.).

  6. Computed Tomography of Prosthetic Heart Valves

    NARCIS (Netherlands)

    Habets, J.

    2012-01-01

    Prosthetic heart valve (PHV) dysfunction is an infrequent but potentially life-threatening disease with a heterogeneous clinical presentation. Patients with PHV dysfunction clinically can present with symptoms of congestive heart failure (dyspnea, fatigue, edema), fever, angina pectoris, dizziness

  7. Successful Thrombolysis of Aortic Prosthetic Valve Thrombosis ...

    African Journals Online (AJOL)

    Arun Kumar Agnihotri

    threatening. Standard surgical treatment using cardiopulmonary bypass carries high maternal and fetal complications. Here we report a case of an antenatal female in first trimester with aortic prosthetic valve thrombosis (PVT), who was successfully ...

  8. DME Prosthetics Orthotics, and Supplies Fee Schedule

    Data.gov (United States)

    U.S. Department of Health & Human Services — Durable Medical Equipment, Prosthetics-Orthotics, and Supplies Fee Schedule. The list contains the fee schedule amounts, floors, and ceilings for all procedure codes...

  9. PLANNING OF IMPLANTO‐PROSTHETIC STRUCTURES IN TOTAL EDENTATION ACCORDING TO BIOMECHANICAL CRITERIA

    Directory of Open Access Journals (Sweden)

    N. VASILE

    2013-07-01

    Full Text Available Scope of the study: Analysis of the biomechanical aspects of the implanto‐prosthetic structures for planning the treatment in cases of total edentation.Materials and method: The study was performed in the Clinics of Prosthetics and Oral Implantology, Military Emergency Hospital of Sibiu, on a group of mandibularly totally edentated patients. Planning of the implanto‐pros‐ thetic treatment had in view the peculiarities of total man‐ dibular edentation and observance of the biomechanical principles. Selection of the type of occlusion restoration considered the presence of parafunctions and the nature of the antagonistic arch. Special attention was paid to the direction and intensity of the forces acting in the region of the future prosthetic work. When bruxism was manifested, its preimplantary removal was compulsory.Results and discussion: In fixed prosthetic restaurati‐ ons applied on implants, distribuition of forces obviously depend on the quality of osteo‐acceptance, as well, and also on the elasticity degree of the prosthetic work. In the case of mobilized prosthetic restorations supported on implants, rigidization of implants from the anterior region of the mandible may be obtained by means of a bar. In decreasing order of their elasticity, the materials employed are: acrylate, composites, noble and seminoble alloys, other metals, ceramics. Ceramics confers maximum stability to implants, yet without redeeming the forces. For amortizing the forces and for a progressive charging of the implants, it is recommended that the first (temporary prosthetic restorations should be made of either acrylate or compo‐ sites.Conclusions. Observance of the occlusological princi‐ ples in cases of occlusal reconstruction represents the determining element which assures the osteo‐acceptance of implants and the integrity of prosthetic restaurations.

  10. Limits of stability in persons with transtibial amputation with respect to prosthetic alignment alterations.

    Science.gov (United States)

    Kolarova, Barbora; Janura, Miroslav; Svoboda, Zdenek; Elfmark, Milan

    2013-11-01

    To evaluate the limits of stability (LOS) in persons with transtibial amputation (TTA), and to determine the effects of prosthetic alignment alterations on motor control strategies. Before-and-after trial. A kinesiology laboratory at a university hospital. Male patients with TTA (n=10) and controls (n=17). Prosthetic alignment. For the LOS test, the maximum excursion, endpoint excursion, direction control, movement velocity, and reaction time with inclination in the forward direction, toward the amputated leg/right leg, and in the backward direction, and toward the nonamputated leg/left leg. Measurements were performed using the following 5 prosthetic alignments: the optimal alignment, with the prosthesis shorter by 1cm, with the prosthesis longer by 1cm, and with the prosthetic foot in 5° of extra plantar flexion and 5° of extra dorsiflexion. Compared with the control group, maximum excursion and direction control were lower (P<.05) in patients with TTA with backward body inclination for all tested prosthetic alignments. Direction control in backward inclination was reduced (P<.05) compared with other tested directions for all assessed prosthetic alignments. Differences between the tested alignments were not significant in any of the tested directions. Patients with TTA have decreased voluntary body inclination backward within the LOS for all tested prosthetic alignments. Compared with controls, changes in prosthetic foot settings by means of rotation in the sagittal plane had a larger impact on movement strategy in patients with TTA than did changes to the length of the prosthesis. Copyright © 2013 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.

  11. Autologous alternative veins may not provide better outcomes than prosthetic conduits for below-knee bypass when great saphenous vein is unavailable.

    Science.gov (United States)

    Avgerinos, Efthymios D; Sachdev, Ulka; Naddaf, Abdallah; Doucet, Dannielle R; Mohapatra, Abhisekh; Leers, Steven A; Chaer, Rabih A; Makaroun, Michel S

    2015-08-01

    There is a need to better define the role of alternative autologous vein (AAV) segments over contemporary prosthetic conduits in patients with critical limb ischemia when great saphenous vein (GSV) is not available for use as the bypass conduit. Consecutive patients who underwent bypass to infrageniculate targets between 2007 and 2011 were categorized in three groups: GSV, AAV, and prosthetic. The primary outcome was graft patency. The secondary outcome was limb salvage. Cox proportional hazards regression was used to adjust for baseline confounding variables. A total of 407 infrainguinal bypasses to below-knee targets were analyzed; 255 patients (63%) received a single-segment GSV, 106 patients (26%) received an AAV, and 46 patients (11%) received a prosthetic conduit. Baseline characteristics were similar among groups, with the exception of popliteal targets and anticoagulation use being more frequent in the prosthetic group. Primary patency at 2 and 5 years was estimated at 47% and 32%, respectively, for the GSV group; 24% and 23% for the AAV group; and 43% and 38% for the prosthetic group. Primary assisted patency at 2 and 5 years was estimated at 71% and 55%, respectively, for the GSV group; 53% and 51% for the AAV group; and 45% and 40% for the prosthetic group. Secondary patency at 2 and 5 years was estimated at 75% and 60%, respectively, for the GSV group; 57% and 55% for the AAV group; and 46% and 41% for the prosthetic group. In Cox analysis, primary patency (hazard ratio [HR], 0.55; P prosthetic grafts except for the primary patency, for which prosthetic was protective (HR, 0.38; P Limb salvage was similar among groups. AAV conduits may not offer a significant patency advantage in midterm follow-up over prosthetic bypasses. Copyright © 2015 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.

  12. Analysis of maxillofacial prosthetics at university dental hospitals in the capital region of Korea.

    Science.gov (United States)

    Kim, Jee-Hwan; Shin, Soo-Yeon; Paek, Janghyun; Lee, Jong-Ho; Kwon, Ho-Beom

    2016-06-01

    The purpose of this study was to investigate the demographic patterns of maxillofacial prosthetic treatment to identify the characteristics and geographic distribution of patients with maxillofacial prosthetics in the capital region of Korea. This retrospective analytical multicenter study was performed by chart reviews. This study included patients who visited the department of prosthodontics at four university dental hospitals for maxillofacial prosthetic rehabilitation. Patients with facial and congenital defects or with insufficient medical data were excluded. The patients were classified into three categories based on the location of the defect. Patients' sex, age, and residential area were analyzed. Pearson's chi-square test with a significance level of 0.05 was used to analyze the variables. Among 540 patients with maxillofacial prosthetics, there were 284 (52.59%) male patients and 256 (47.41%) female patients. The number of the patients varied greatly by hospital. Most patients were older than 70, and the most common defect was a hard palate defect. Chi-square analysis did not identify any significant differences in sex, age, and distance to hospital for any defect group (P>.05). The results of this study indicated that there was imbalance in the distribution of patients with maxillofacial prosthetic among the hospitals in the capital region of Korea. Considerations on specialists and insurance policies for the improvement of maxillofacial prosthetics in Korea are required.

  13. Incidence of Prosthetic Complications associated with Implant-borne Prosthesis in a Sleep Disorder Center.

    Science.gov (United States)

    Suneel, Venkatesh B; Kotian, Santhosh; Jujare, Ravikanth H; Shetty, Adarsh K; Nidhi, Sneh; Grover, Shehkar

    2017-09-01

    Obstructive sleep apnea (OSA) is one of the common prevalent conditions present worldwide. The process of abnormal habits related to clenching and grinding of teeth is referred to as bruxism and is characterized under the heading of parafunctional activity of the masticatory system. Osseointegrated dental implants represent advancements in the field of odontology. Despite its high success rate, failure and complications are often associated with dental implant treatment due to a number of factors. Hence, we aimed for the present study to assess the incidence of prosthetic complications in patients rehabilitated with implant-borne prosthesis in a sleep disorder unit. The present study included the assessment of all the patients who underwent prosthetic rehabilitation by dental implants. An experienced registered prosthodontist was given duty for examination of all the cases from the record file data. Prosthetic complications in the patients were identified using photographs, radiographs, and all other relevant data of the patients obtained from the record files. All types of complications and other factors were recorded separately and analyzed. While correlating the prosthetic complications in OSA patients grouped based on number of dental implants, nonsignificant results were obtained. Significant correlation was observed while comparing the prosthetic complications divided based on type of prosthesis. Fracture of the porcelain was observed in four and eight cases respectively, of screwed and cemented dental implant cases. Some amount of significant correlation existed between the incidences of prosthetic complications and OSA. Proper history of the patients undergoing dental implant procedures should be taken to avoid failure.

  14. Surgical and Prosthetic Rehabilitation of Combination Syndrome

    Directory of Open Access Journals (Sweden)

    Paolo Carlino

    2014-01-01

    Full Text Available The aim of this report is to analyze the clinical symptoms, ethologic factors, and prosthetic rehabilitation in a case of Combination Syndrome (CS. The treatment of CS can be conventional or surgical, with or without the bone reconstruction of maxilla. The correct prosthetic treatment helps this kind of patients to restore the physiologic occlusion plane to allow a correct masticatory and aesthetic function. Management of this kind of patients can be a challenge for a dental practitioner.

  15. Prosthetic Mesh Repair for Incarcerated Inguinal Hernia

    Directory of Open Access Journals (Sweden)

    Cihad Tatar

    2016-08-01

    Full Text Available Background: Incarcerated inguinal hernia is a commonly encountered urgent surgical condition, and tension-free repair is a well-established method for the treatment of noncomplicated cases. However, due to the risk of prosthetic material-related infections, the use of mesh in the repair of strangulated or incarcerated hernia has often been subject to debate. Recent studies have demonstrated that biomaterials represent suitable materials for performing urgent hernia repair. Certain studies recommend mesh repair only for cases where no bowel resection is required; other studies, however, recommend mesh repair for patients requiring bowel resection as well. Aim: The aim of this study was to compare the outcomes of different surgical techniques performed for strangulated hernia, and to evaluate the effect of mesh use on postoperative complications. Study Design: Retrospective cross-sectional study. Methods: This retrospective study was performed with 151 patients who had been admitted to our hospital’s emergency department to undergo surgery for a diagnosis of incarcerated inguinal hernia. The patients were divided into two groups based on the applied surgical technique. Group 1 consisted of 112 patients treated with mesh-based repair techniques, while Group 2 consisted of 39 patients treated with tissue repair techniques. Patients in Group 1 were further divided into two sub-groups: one consisting of patients undergoing bowel resection (Group 3, and the other consisting of patients not undergoing bowel resection (Group 4. Results: In Group 1, it was observed that eight (7.14% of the patients had wound infections, while two (1.78% had hematomas, four (3.57% had seromas, and one (0.89% had relapse. In Group 2, one (2.56% of the patients had a wound infection, while three (7.69% had hematomas, one (2.56% had seroma, and none had relapses. There were no statistically significant differences between the two groups with respect to wound infection

  16. Heme Distortions in Sperm-Whale Carbonmonoxy Myoglobin: Correlations between Rotational Strengths and Heme Distortions in MD-Generated Structures

    Energy Technology Data Exchange (ETDEWEB)

    KIEFL,CHRISTOPH; SCREERAMA,NARASIMHA; LU,YI; QIU,YAN; SHELNUTT,JOHN A.; WOODY,ROBERT W.

    2000-07-13

    The authors have investigated the effects of heme rotational isomerism in sperm-whale carbonmonoxy myoglobin using computational techniques. Several molecular dynamics simulations have been performed for the two rotational isomers A and B, which are related by a 180{degree} rotation around the {alpha}-{gamma} axis of the heme, of sperm-whale carbonmonoxy myoglobin in water. Both neutron diffraction and NMR structures were used as starting structures. In the absence of an experimental structure, the structure of isomer B was generated by rotating the heme in the structure of isomer A. Distortions of the heme from planarity were characterized by normal coordinate structural decomposition and by the angle of twist of the pyrrole rings from the heme plane. The heme distortions of the neutron diffraction structure were conserved in the MD trajectories, but in the NMR-based trajectories, where the heme distortions are less well defined, they differ from the original heme deformations. The protein matrix induced similar distortions on the heroes in orientations A and B. The results suggest that the binding site prefers a particular macrocycle conformation, and a 180{degree} rotation of the heme does not significantly alter the protein's preference for this conformation. The intrinsic rotational strengths of the two Soret transitions, separated according to their polarization in the heme plane, show strong correlations with the ruf-deformation and the average twist angle of the pyrrole rings. The total rotational strength, which includes contributions from the chromophores in the protein, shows a weaker correlation with heme distortions.

  17. Heme acquisition mechanisms of Porphyromonas gingivalis - strategies used in a polymicrobial community in a heme-limited host environment.

    Science.gov (United States)

    Smalley, J W; Olczak, T

    2017-02-01

    Porphyromonas gingivalis, a main etiologic agent and key pathogen responsible for initiation and progression of chronic periodontitis requires heme as a source of iron and protoporphyrin IX for its survival and the ability to establish an infection. Porphyromonas gingivalis is able to accumulate a defensive cell-surface heme-containing pigment in the form of μ-oxo bisheme. The main sources of heme for P. gingivalis in vivo are hemoproteins present in saliva, gingival crevicular fluid, and erythrocytes. To acquire heme, P. gingivalis uses several mechanisms. Among them, the best characterized are those employing hemagglutinins, hemolysins, and gingipains (Kgp, RgpA, RgpB), TonB-dependent outer-membrane receptors (HmuR, HusB, IhtA), and hemophore-like proteins (HmuY, HusA). Proteins involved in intracellular heme transport, storage, and processing are less well characterized (e.g. PgDps). Importantly, P. gingivalis may also use the heme acquisition systems of other bacteria to fulfill its own heme requirements. Porphyromonas gingivalis displays a novel paradigm for heme acquisition from hemoglobin, whereby the Fe(II)-containing oxyhemoglobin molecule must first be oxidized to methemoglobin to facilitate heme release. This process not only involves P. gingivalis arginine- and lysine-specific gingipains, but other proteases (e.g. interpain A from Prevotella intermedia) or pyocyanin produced by Pseudomonas aeruginosa. Porphyromonas gingivalis is then able to fully proteolyze the more susceptible methemoglobin substrate to release free heme or to wrest heme from it directly through the use of the HmuY hemophore. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. Dietary heme-mediated PPARα activation does not affect the heme-induced epithelial hyperproliferation and hyperplasia in mouse colon.

    Directory of Open Access Journals (Sweden)

    Noortje Ijssennagger

    Full Text Available Red meat consumption is associated with an increased colon cancer risk. Heme, present in red meat, injures the colon surface epithelium by luminal cytotoxicity and reactive oxygen species. This surface injury is overcompensated by hyperproliferation and hyperplasia of crypt cells. Transcriptome analysis of mucosa of heme-fed mice showed, besides stress- and proliferation-related genes, many upregulated lipid metabolism-related PPARα target genes. The aim of this study was to investigate the role of PPARα in heme-induced hyperproliferation and hyperplasia. Male PPARα KO and WT mice received a purified diet with or without heme. As PPARα is proposed to protect against oxidative stress and lipid peroxidation, we hypothesized that the absence of PPARα leads to more surface injury and crypt hyperproliferation in the colon upon heme-feeding. Heme induced luminal cytotoxicity and lipid peroxidation and colonic hyperproliferation and hyperplasia to the same extent in WT and KO mice. Transcriptome analysis of colonic mucosa confirmed similar heme-induced hyperproliferation in WT and KO mice. Stainings for alkaline phosphatase activity and expression levels of Vanin-1 and Nrf2-targets indicated a compromised antioxidant defense in heme-fed KO mice. Our results suggest that the protective role of PPARα in antioxidant defense involves the Nrf2-inhibitor Fosl1, which is upregulated by heme in PPARα KO mice. We conclude that PPARα plays a protective role in colon against oxidative stress, but PPARα does not mediate heme-induced hyperproliferation. This implies that oxidative stress of surface cells is not the main determinant of heme-induced hyperproliferation and hyperplasia.

  19. Dioxygen reactivity of meso-hydroxylated hemes: intermediates in ...

    Indian Academy of Sciences (India)

    464. Scheme 2. in which the HO-bound heme could be converted to verdoheme and biliverdin by ascorbic acid or hydra- zine in the absence of NADPH and cytochrome. P450 reductase via the formation of similar interme- diates observed during the biological oxidation of heme by HO.2–7 The process involves initial meso.

  20. Heme oxygenase-1 and carbon monoxide in pulmonary medicine

    NARCIS (Netherlands)

    Slebos, DJ; Ryter, SW; Choi, AMK

    2003-01-01

    Heme oxygenase-1 (HO-1), an inducible stress protein, confers cytoprotection against oxidative stress in vitro and in vivo. In addition to its physiological role in heme degradation, HO-1 may influence a number of cellular processes, including growth, inflammation, and apoptosis. By virtue of

  1. Structures of the substrate-free and product-bound forms of HmuO, a heme oxygenase from corynebacterium diphtheriae: x-ray crystallography and molecular dynamics investigation.

    Science.gov (United States)

    Unno, Masaki; Ardèvol, Albert; Rovira, Carme; Ikeda-Saito, Masao

    2013-11-29

    Heme oxygenase catalyzes the degradation of heme to biliverdin, iron, and carbon monoxide. Here, we present crystal structures of the substrate-free, Fe(3+)-biliverdin-bound, and biliverdin-bound forms of HmuO, a heme oxygenase from Corynebacterium diphtheriae, refined to 1.80, 1.90, and 1.85 Å resolution, respectively. In the substrate-free structure, the proximal and distal helices, which tightly bracket the substrate heme in the substrate-bound heme complex, move apart, and the proximal helix is partially unwound. These features are supported by the molecular dynamic simulations. The structure implies that the heme binding fixes the enzyme active site structure, including the water hydrogen bond network critical for heme degradation. The biliverdin groups assume the helical conformation and are located in the heme pocket in the crystal structures of the Fe(3+)-biliverdin-bound and the biliverdin-bound HmuO, prepared by in situ heme oxygenase reaction from the heme complex crystals. The proximal His serves as the Fe(3+)-biliverdin axial ligand in the former complex and forms a hydrogen bond through a bridging water molecule with the biliverdin pyrrole nitrogen atoms in the latter complex. In both structures, salt bridges between one of the biliverdin propionate groups and the Arg and Lys residues further stabilize biliverdin at the HmuO heme pocket. Additionally, the crystal structure of a mixture of two intermediates between the Fe(3+)-biliverdin and biliverdin complexes has been determined at 1.70 Å resolution, implying a possible route for iron exit.

  2. Cetacean Swimming with Prosthetic Limbs

    Science.gov (United States)

    Bode-Oke, Ayodeji; Ren, Yan; Dong, Haibo; Fish, Frank

    2016-11-01

    During entanglement in fishing gear, dolphins can suffer abrasions and amputations of flukes and fins. As a result, if the dolphin survives the ordeal, swimming performance is altered. Current rehabilitation technques is the use of prosthesis to regain swimming ability. In this work, analyses are focused on two dolphins with locomotive impairment; Winter (currently living in Clearwater Marine Aquarium in Florida) and Fuji (lived in Okinawa Churaumi Aquarium in Japan). Fuji lost about 75% of its fluke surface to necrosis (death of cells) and Winter lost its tail due to amputation. Both dolphins are aided by prosthetic tails that mimic the shape of a real dolphin tail. Using 3D surface reconstruction techniques and a high fidelity Computational Fluid Dynamics (CFD) flow solver, we were able to elucidate the kinematics and hydrodynamics and fluke deformation of these swimmers to clarify the effectiveness of prostheses in helping the dolphins regain their swimming ability. Associated with the performance, we identified distinct features in the wake structures that can explain this gap in the performance compared to a healthy dolphin. This work was supported by ONR MURI Grant Number N00014-14-1-0533.

  3. Functional characterization of the Shigella dysenteriae heme ABC transporter.

    Science.gov (United States)

    Burkhard, Kimberly A; Wilks, Angela

    2008-08-05

    The heme ATP binding cassette (ABC) transporter, ShuUV, of Shigella dysenteriae has been incorporated into proteoliposomes. Functional characterization of ShuUV revealed that ATP hydrolysis and transport of heme from the periplasmic binding protein, ShuT, to the cytoplasmic binding protein, ShuS, are coupled. Site-directed mutagenesis of ShuT residues proposed to be required for stabilization of the complex abolished heme transport. Furthermore, residues His-252 and His-262, located in the translocation channel of ShuU, were required for the release of heme from ShuT and translocation to ShuS. The initial functional characterization of an in vitro heme uptake system provides a platform for future spectroscopic studies.

  4. Kinematics in the terminal swing phase of unilateral transfemoral amputees: microprocessor-controlled versus swing-phase control prosthetic knees.

    Science.gov (United States)

    Mâaref, Khaled; Martinet, Noël; Grumillier, Constance; Ghannouchi, Slaheddine; André, Jean Marie; Paysant, Jean

    2010-06-01

    To analyze the spatiotemporal parameters in the terminal swing phase of the prosthetic limb in unilateral transfemoral amputees (TFAs) compared with a group of asymptomatic subjects, and to identify a latency period (LP) in the TFA between the full extension of the prosthetic knee and the initial ground contact of the ipsilateral foot. To study the correlation between the LP and the duration of the swing phase. To evaluate the influence of the type of knee, the time since amputation, and the amputation level on the latency period. Three-dimensional gait analysis with an optoelectronic device. Gait analysis laboratory of a re-education and functional rehabilitation service. TFA (n=29) and able-bodied (n=15) subjects. Not applicable. Spatiotemporal and kinematics gait parameters. The swing phase and the LP of the prosthetic limb, associated with a consequently longer single-limb stance phase in the intact limb, were significantly longer than those measured in the intact limbs of these subjects, as well as those measured on both lower limbs of the able-bodied subjects (Pprosthetic side. The LP measured in the prosthetic limb of TFA with a swing-phase control prosthetic knee is significantly greater than in those using the microprocessor-controlled prosthetic knee (Plimb of TFA, the LP is significantly greater in the prosthetic limb. It can explain the lengthened swing phase on the prosthetic side of those subjects. The use of a microprocessor-controlled prosthetic knee allows the LP to be reduced. This LP appears to be necessary to insure the stability of the prosthetic knee. We suggest calling this time "confidence time." Copyright 2010 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.

  5. Increased Plasma Levels of Heme Oxygenase-1 in Human Brucellosis.

    Science.gov (United States)

    Chen, Zhe; Zhang, Yu-Xue; Fu, Dong-Wei; Gao, Qing-Feng; Ge, Feng-Xia; Liu, Wei-Hua

    2016-08-01

    Brucellosis is associated with inflammation and the oxidative stress response. Heme oxygenase-1 (HO-1) is a cytoprotective stress-responsive enzyme that has anti-inflammatory and anti-oxidant effects. Nevertheless, the role of HO-1 in human brucellosis has not yet been studied. The aim of this study was to examine the plasma levels of HO-1 in patients with brucellosis and to evaluate the ability of plasma HO-1 levels as an auxiliary diagnosis, a severity predictor, and a monitor for brucellosis treatments. A total of 75 patients with brucellosis were divided into the acute, subacute, chronic active, and chronic stable groups. An additional 20 volunteers were included as the healthy control group. The plasma HO-1 levels and other laboratory parameters were measured in all groups. Furthermore, the plasma levels of HO-1 in the acute group were compared before and after treatment. The plasma HO-1 levels were considerably increased in the acute (4.97 ± 3.55), subacute (4.98 ± 3.23), and chronic active groups (4.43 ± 3.00) with brucellosis compared to the healthy control group (1.03 ± 0.63) (p brucellosis (r = 0.707, p brucellosis status and may be used as a supplementary plasma marker for diagnosing brucellosis and monitoring its treatment.

  6. Percutaneous management of prosthetic valve thrombosis.

    Science.gov (United States)

    Hariram, Vuppaladadhiam

    2014-01-01

    Thrombosis of a prosthetic valve is a serious complication in patients with prosthetic heart valves. Thrombolysis is the initial choice of treatment. Patients who do not respond to thrombolysis are subjected to surgery which carries a high risk. We report a case series of 5 patients with prosthetic mitral valve thrombosis who did not respond to thrombolysis and were subjected to percutaneous manipulation of the prosthetic valves successfully and improved. Five patients who were diagnosed to have prosthetic mitral valve thrombosis and failed to respond to a minimum of 36 h of thrombolysis (persistent symptoms with increased gradients, abnormal findings on fluoroscopy),were subjected to percutaneous treatment after receiving proper consent. None of them had a visible thrombus on transthoracic echocardiogram. All patients underwent transseptal puncture following which a 6F JR4 guiding catheter was passed into the left atrium. The valve leaflets were repeatedly hit gently under fluoroscopic guidance till they regained their normal mobility. Mean age was 38.8 years. Average peak and mean gradients prior to the procedure were 38 and 25 and after the procedure were 12 and 6 mm of Hg respectively. All patients had successful recovery of valve motion on fluoroscopy with normalization of gradients and complete resolution of symptoms. None of the patients had any focal neurological deficits, embolic manifestations or bleeding complications. Percutaneous manipulation of prosthetic valves in selected patients with prosthetic valve thrombosis who do not respond to thrombolytic therapy is feasible and can be used as an alternative to surgery. Copyright © 2014 Cardiological Society of India. Published by Elsevier B.V. All rights reserved.

  7. Reconsidering evidence-based practice in prosthetic rehabilitation: a shared enterprise.

    Science.gov (United States)

    van Twillert, Sacha; Geertzen, Jan; Hemminga, Titia; Postema, Klaas; Lettinga, Ant

    2013-06-01

    A divide is experienced between producers and users of evidence in prosthetic rehabilitation. To discuss the complexity inherent in establishing evidence-based practice in a prosthetic rehabilitation team illustrated by the case of prosthetic prescription for elderly dysvascular transfemoral amputee patients. A qualitative research design was used, in which data from multiple sources was triangulated to extract themes for discussion. This discussion paper draws on empirical material gathered by individual and focus-group interviews with members of a prosthetic rehabilitation team, information on technological advancements presented on websites of orthopaedic industry, guidelines and literature study. A prosthetic rehabilitation team needs to deal with lack of evidence, contradictory results, various classification systems, diverging interests of different stakeholders and many modifying factors, and all of this in a continuous technological changing environment. Integrating research designs with different strengths but not sharing the same biases may help researchers to deal with the multimorbidity and multifaceted disability of the target group. Articulating clinical knowledge, patients' needs and values in a systematic way provides depth, detail, nuance and context for evidence-based practice issues in prosthetic rehabilitation. Reconsidering the relationship between evidence, technology and rehabilitation practice is an imperative shared enterprise for clinicians and researchers. Scientific, clinical and patient-related knowledge are seen as important knowledge practices that should inform and strengthen each other. This discussion paper puts the academic clinical debate on evidence-based practice in prosthetics and orthotics in another light. By demonstrating the complexities surrounding evidence-based practice, it is argued and illustrated how both researchers and clinicians can contribute to optimal patient care in which evidence, technology and

  8. Prosthetic fitting after rotationplasty of the knee.

    Science.gov (United States)

    So, Noel F; Andrews, Karen L; Anderson, Kimberly; Gozola, Michael A; Shives, Thomas C; Rose, Peter S; Shaughnessy, William J; Sim, Franklin H

    2014-04-01

    The objectives of this study were to describe the authors' experience with the timeline for prosthetic fitting after rotationplasty of the knee and to determine factors that may be associated with earlier prosthetic fitting. The authors conducted a retrospective observational study of 12 patients who underwent rotationplasty of the knee and received prosthetic care at this institution. All patients had oncologic causes for surgery. The median age at amputation was 10 yrs. The overall survival rate was 92%. Five patients received a preliminary bypass prosthesis. All 12 patients were successfully fitted with a definitive prosthesis. Three patients were fitted within 90 days; two of these three patients did not require chemotherapy. The median time for definitive prosthetic fitting in the ten patients requiring chemotherapy was 230.5 days (range, 85-425 days). Nine patients had documentation supporting a return to sport/premorbid physical recreational activities. In the authors' experience, chemotherapy was associated with delayed definitive prosthetic fitting. Typically, the patients who required rotationplasty for cancer completed fitting with a definitive prosthesis in 6 mos. The findings of this study validate previous reports and confirm that most rotationplasty patients have excellent outcomes with return to premorbid physical activities.

  9. Heme oxygenase-1 end-products carbon monoxide and biliverdin ameliorate murine collagen induced arthritis.

    Science.gov (United States)

    Bonelli, M; Savitskaya, A; Steiner, C-W; Rath, E; Bilban, M; Wagner, O; Bach, F H; Smolen, J S; Scheinecker, C

    2012-01-01

    Heme oxygenase-1 (HO-1) which degrades Heme to free iron, biliverdin and carbon monoxide (CO) plays an important role in inflammation. There are, however, conflicting data concerning the role of HO-1 in rheumatoid arthritis (RA) and the therapeutic potential of individual heme degradation products remains to be determined. We therefore investigated the effect of CO and biliverdin upon therapeutic administration in the murine collagen induced arthritis (CIA) model of RA. CIA was induced in DBA/1 mice. Anti-CII antibody levels were determined by ELISA. Mice were scored for paw swelling and grip strength. After the first clinical signs of arthritis one group of animals was treated with biliverdin, the second group was treated with CO. After 60 days all animals were sacrificed and analysed for histomorphological signs of arthritis. All animals immunised with CII developed serum anti-CII antibodies. Antibody levels were decreased in the CO-treated group. Both, Biliverdin and the CO-treated animals, showed an improvement in clinical disease activity. Histological analysis revealed significantly less inflammation, erosion and reduced numbers of osteoclasts in CO-treated animals only, whereas cartilage degradation was prevented in both biliverdin and CO-treated animals. Our data demonstrate a beneficial effect of CO, in particular, and biliverdin, on inflammation and bone destruction in the CIA mouse model.

  10. The mitochondrial heme exporter FLVCR1b mediates erythroid differentiation.

    Science.gov (United States)

    Chiabrando, Deborah; Marro, Samuele; Mercurio, Sonia; Giorgi, Carlotta; Petrillo, Sara; Vinchi, Francesca; Fiorito, Veronica; Fagoonee, Sharmila; Camporeale, Annalisa; Turco, Emilia; Merlo, Giorgio R; Silengo, Lorenzo; Altruda, Fiorella; Pinton, Paolo; Tolosano, Emanuela

    2012-12-01

    Feline leukemia virus subgroup C receptor 1 (FLVCR1) is a cell membrane heme exporter that maintains the balance between heme levels and globin synthesis in erythroid precursors. It was previously shown that Flvcr1-null mice died in utero due to a failure of erythropoiesis. Here, we identify Flvcr1b, a mitochondrial Flvcr1 isoform that promotes heme efflux into the cytoplasm. Flvcr1b overexpression promoted heme synthesis and in vitro erythroid differentiation, whereas silencing of Flvcr1b caused mitochondrial heme accumulation and termination of erythroid differentiation. Furthermore, mice lacking the plasma membrane isoform (Flvcr1a) but expressing Flvcr1b had normal erythropoiesis, but exhibited hemorrhages, edema, and skeletal abnormalities. Thus, FLVCR1b regulates erythropoiesis by controlling mitochondrial heme efflux, whereas FLVCR1a expression is required to prevent hemorrhages and edema. The aberrant expression of Flvcr1 isoforms may play a role in the pathogenesis of disorders characterized by an imbalance between heme and globin synthesis.

  11. Isoporphyrin Intermediate in Heme Oxygenase Catalysis

    Science.gov (United States)

    Evans, John P.; Niemevz, Fernando; Buldain, Graciela; de Montellano, Paul Ortiz

    2008-01-01

    Human heme oxygenase-1 (hHO-1) catalyzes the O2- and NADPH-dependent oxidation of heme to biliverdin, CO, and free iron. The first step involves regiospecific insertion of an oxygen atom at the α-meso carbon by a ferric hydroperoxide and is predicted to proceed via an isoporphyrin π-cation intermediate. Here we report spectroscopic detection of a transient intermediate during oxidation by hHO-1 of α-meso-phenylheme-IX, α-meso-(p-methylphenyl)-mesoheme-III, and α-meso-(p-trifluoromethylphenyl)-mesoheme-III. In agreement with previous experiments (Wang, J., Niemevz, F., Lad, L., Huang, L., Alvarez, D. E., Buldain, G., Poulos, T. L., and Ortiz de Montellano, P. R. (2004) J. Biol. Chem. 279, 42593–42604), only the α-biliverdin isomer is produced with concomitant formation of the corresponding benzoic acid. The transient intermediate observed in the NADPH-P450 reductase-catalyzed reaction accumulated when the reaction was supported by H2O2 and exhibited the absorption maxima at 435 and 930 nm characteristic of an isoporphyrin. Product analysis by reversed phase high performance liquid chromatography and liquid chromatography electrospray ionization mass spectrometry of the product generated with H2O2 identified it as an isoporphyrin that, on quenching, decayed to benzoylbiliverdin. In the presence of H218O2, one labeled oxygen atom was incorporated into these products. The hHO-1-isoporphyrin complexes were found to have half-lives of 1.7 and 2.4 h for the p-trifluoromethyl- and p-methyl-substituted phenylhemes, respectively. The addition of NADPH-P450 reductase to the H2O2-generated hHO-1-isoporphyrin complex produced α-biliverdin, confirming its role as a reaction intermediate. Identification of an isoporphyrin intermediate in the catalytic sequence of hHO-1, the first such intermediate observed in hemoprotein catalysis, completes our understanding of the critical first step of heme oxidation. PMID:18487208

  12. Comparison of patient-reported outcomes after traumatic upper extremity amputation: Replantation versus prosthetic rehabilitation.

    Science.gov (United States)

    Pet, Mitchell A; Morrison, Shane D; Mack, Jacob S; Sears, Erika D; Wright, Thomas; Lussiez, Alisha D; Means, Kenneth R; Higgins, James P; Ko, Jason H; Cederna, Paul S; Kung, Theodore A

    2016-12-01

    After major upper extremity traumatic amputation, replantation is attempted based upon the assumption that outcomes for a replanted limb exceed those for revision amputation with prosthetic rehabilitation. While some reports have examined functional differences between these patients, it is increasingly apparent that patient perceptions are also critical determinants of success. Currently, little patient-reported outcomes data exists to support surgical decision-making in the setting of major upper extremity traumatic amputation. Therefore, the purpose of this study is to directly compare patient-reported outcomes after replantation versus prosthetic rehabilitation. At three tertiary care centers, patients with a history of traumatic unilateral upper extremity amputation at or between the radiocarpal and elbow joints were identified. Patients who underwent either successful replantation or revision amputation with prosthetic rehabilitation were contacted. Patient-reported health status was evaluated with both DASH and MHQ instruments. Intergroup comparisons were performed for aggregate DASH score, aggregate MHQ score on the injured side, and each MHQ domain. Nine patients with successful replantation and 22 amputees who underwent prosthetic rehabilitation were enrolled. Aggregate MHQ score for the affected extremity was significantly higher for the Replantation group compared to the Prosthetic Rehabilitation group (47.2 vs. 35.1, pprosthetic rehabilitation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. Determining 1-Yr Prosthetic Use for Mobility Prognoses for Community-Dwelling Adults with Lower-Limb Amputation: Development of a Clinical Prediction Rule.

    Science.gov (United States)

    Wong, Christopher Kevin; Young, Rebecca S; Ow-Wing, Carlyn; Karimi, Parisa

    2016-05-01

    The objective of this study was to develop a prognostic clinical prediction rule to identify people not achieving community walking level prosthetic use after 1 yr. This is a prospective longitudinal cohort study of community-dwelling adults with lower-limb amputations recruited from support groups and prosthetic clinics. Participants completed Activities-specific Balance Confidence and Houghton prosthetic use for mobility self-report scales and the Berg Balance Scale. The clinical prediction rule was developed using multivariate logistic regression, receiver operating curves, and probability statistics to identify people not achieving community walking level prosthetic use (Houghton scores prosthetic walking level with excellent accuracy (area under the curve >0.96) using four criteria: initial Houghton, Activities-specific Balance Confidence, and Berg Balance Scale tasks 9 (retrieve object from floor) and 10 (look behind over shoulders). Failure to exceed cutoff scores in two or more criteria yielded posttest probability of not reaching community walking prosthetic use 1 yr later for 90% of participants or higher. Accurate 1-yr prosthetic use for mobility prognoses can be obtained by screening prosthetic use, balance confidence, and balance ability to identify community-dwelling people with lower-limb amputations unlikely to achieve community walking prosthetic use.

  14. Heme synthase (ferrochelatase) catalyzes the removal of iron from heme and demetalation of metalloporphyrins.

    Science.gov (United States)

    Taketani, Shigeru; Ishigaki, Mutsumi; Mizutani, Atsushi; Uebayashi, Masashi; Numata, Masahiro; Ohgari, Yoshiko; Kitajima, Sakihito

    2007-12-25

    The red pigments in meat products, including cooked cured ham, arise from the reaction of myoglobin with nitric oxide generated from exogenous nitrite. Since carcinogenic nitrosoamines may be generated by the treatment of meats with nitrite, the production of nitrite-free meat products is an attractive alternative. Raw dry-cured (Parma) hams are produced by the treatment of meats with salts other than nitrite. Analysis of pigments in raw dry-cured hams reveals that the main pigment is zinc protoporphyrin, suggesting that the conversion of heme to zinc protoporphyrin occurs via an iron-removal reaction from myoglobin heme during the processing of raw hams. Purification of the iron-removal enzyme showed that it was identical to ferrochelatase. Recombinant ferrochelatase in combination with NADH-cytochrome b5 reductase catalyzed NADH-dependent iron-removal reaction from hemin and hemoproteins. Metal ions such as zinc and cobalt were also removed from the corresponding metalloporphyrins. The addition of zinc ions led to the formation of zinc protoporphyrin. In cultured cells, the conversion of zinc mesoporphyrin to mesoheme was observed to be dependent on ferrochelatase and could be markedly induced during erythroid differentiation. This is the first demonstration of a new enzyme reaction, the reverse reaction of ferrochelatase, which may contribute to a new route of the recycling of protoporphyrin and heme in cells.

  15. The effect of proteins from animal source foods on heme iron bioavailability in humans.

    Science.gov (United States)

    Pizarro, Fernando; Olivares, Manuel; Valenzuela, Carolina; Brito, Alex; Weinborn, Valerie; Flores, Sebastián; Arredondo, Miguel

    2016-04-01

    Forty-five women (35-45 year) were randomly assigned to three iron (Fe) absorption sub-studies, which measured the effects of dietary animal proteins on the absorption of heme Fe. Study 1 was focused on heme, red blood cell concentrate (RBCC), hemoglobin (Hb), RBCC+beef meat; study 2 on heme, heme+fish, chicken, and beef; and study 3 on heme and heme+purified animal protein (casein, collagen, albumin). Study 1: the bioavailability of heme Fe from Hb was similar to heme only (∼13.0%). RBCC (25.0%) and RBCC+beef (21.3%) were found to be increased 2- and 1.6-fold, respectively, when compared with heme alone (pProteins from animal source foods and their digestion products did not enhance heme Fe absorption. Copyright © 2015. Published by Elsevier Ltd.

  16. Contribution of prosthetic treatment considerations for dental extractions of permanent teeth

    Science.gov (United States)

    Fernández-Barrera, Miguel Ángel; Casanova-Rosado, Juan Fernando; Mendoza-Rodríguez, Martha; Escoffié-Ramírez, Mauricio; Casanova-Rosado, Alejandro José; Navarrete-Hernández, José de Jesús; Maupomé, Gerardo

    2016-01-01

    Background. Tooth loss is an easily identifiable outcome that summarizes a complex suite of factors in an individual’s history of dental disease and its treatment by dental services over a lifetime. Assessment of overall tooth loss data is essential for epidemiologically evaluating the adequacy of dental care provided at a systems level, as well as for placing in context tooth loss for non-disease causes. For example, when derived from prosthetic treatment planning, the latter may unfortunately lead to some teeth being extracted (pulled) for the sake of better comprehensive clinical results. The objective of the present manuscript was to identify the contribution to overall tooth loss, by extraction of permanent teeth because of prosthetic treatment reasons. Material and Methods. A cross-sectional study included sex, age, total number of extractions performed by subject, sextant (anterior vs. posterior), group of teeth (incisors, canines, premolars and molars), upper or lower arch, and the main reason underlying extraction (extraction for any reason vs. prosthetic treatment), in patients 18 years of age and older seeking care at a dental school clinic in Mexico. A multivariate logistic regression model was generated. Results. A total of 749 teeth were extracted in 331 patients; 161 teeth (21.5% of total) were extracted for explicit prosthetic treatment indications. As age increased, the likelihood of having an extraction for prosthetic reasons increased 3% (OR = 1.03, p extractions increased, the risk of having an extraction for prosthetic reasons decreased (OR = 0.94, p extractions of permanent teeth were performed for prosthetic reasons in this dental school clinical environment; age, sex, type of tooth, and the total number of extractions moderated such pattern. PMID:27441103

  17. Can We Achieve Intuitive Prosthetic Elbow Control Based on Healthy Upper Limb Motor Strategies?

    Directory of Open Access Journals (Sweden)

    Manelle Merad

    2018-02-01

    Full Text Available Most transhumeral amputees report that their prosthetic device lacks functionality, citing the control strategy as a major limitation. Indeed, they are required to control several degrees of freedom with muscle groups primarily used for elbow actuation. As a result, most of them choose to have a one-degree-of-freedom myoelectric hand for grasping objects, a myoelectric wrist for pronation/supination, and a body-powered elbow. Unlike healthy upper limb movements, the prosthetic elbow joint angle, adjusted prior to the motion, is not involved in the overall upper limb movements, causing the rest of the body to compensate for the lack of mobility of the prosthesis. A promising solution to improve upper limb prosthesis control exploits the residual limb mobility: like in healthy movements, shoulder and prosthetic elbow motions are coupled using inter-joint coordination models. The present study aims to test this approach. A transhumeral amputated individual used a prosthesis with a residual limb motion-driven elbow to point at targets. The prosthetic elbow motion was derived from IMU-based shoulder measurements and a generic model of inter-joint coordinations built from healthy individuals data. For comparison, the participant also performed the task while the prosthetic elbow was implemented with his own myoelectric control strategy. The results show that although the transhumeral amputated participant achieved the pointing task with a better precision when the elbow was myoelectrically-controlled, he had to develop large compensatory trunk movements. Automatic elbow control reduced trunk displacements, and enabled a more natural body behavior with synchronous shoulder and elbow motions. However, due to socket impairments, the residual limb amplitudes were not as large as those of healthy shoulder movements. Therefore, this work also investigates if a control strategy whereby prosthetic joints are automatized according to healthy individuals

  18. Absorption by Isolated Ferric Heme Nitrosyl Cations In Vacuo

    DEFF Research Database (Denmark)

    Wyer, Jean; Nielsen, Steen Brøndsted

    2012-01-01

    Keywords:biophysics;gas-phase spectroscopy;heme proteins;mass spectrometry;nitric oxide Almost innocent: In photobiophysical studies of ferric heme nitrosyl complexes, the absorption spectra of six-coordinate complexes with NO and Met or Cys are similar to that of the five-coordinate complex ion ......(heme)(NO)+. Since the absorption spectra of related proteins with histidine as the proximal ligand are similar to those of the gaseous complexes, the protein microenvironment has little effect on the lowest-energy transition of the porphyrin macrocycle....

  19. Discovery of Intracellular Heme-binding Protein HrtR, Which Controls Heme Efflux by the Conserved HrtB-HrtA Transporter in Lactococcus lactis*

    Science.gov (United States)

    Lechardeur, Delphine; Cesselin, Bénédicte; Liebl, Ursula; Vos, Marten H.; Fernandez, Annabelle; Brun, Célia; Gruss, Alexandra; Gaudu, Philippe

    2012-01-01

    Most commensal and food bacteria lack heme biosynthesis genes. For several of these, the capture of environmental heme is a means of activating aerobic respiration metabolism. Our previous studies in the Gram-positive bacterium Lactococcus lactis showed that heme exposure strongly induced expression of a single operon, called here hrtRBA, encoding an ortholog of the conserved membrane hrt (heme-regulated transporter) and a unique transcriptional regulator that we named HrtR. We show that HrtR expressed as a fusion protein is a heme-binding protein. Heme iron interaction with HrtR is non-covalent, hexacoordinated, and involves two histidines, His-72 and His-149. HrtR specifically binds a 15-nt palindromic sequence in the hrtRBA promoter region, which is needed for hrtRBA repression. HrtR-DNA binding is abolished by heme addition, which activates expression of the HrtB-HrtA (HrtBA) transporter in vitro and in vivo. The use of HrtR as an intracellular heme sensor appears to be conserved among numerous commensal bacteria, in contrast with numerous Gram-positive pathogens that use an extracellular heme-sensing system, HssRS, to regulate hrt. Finally, we show for the first time that HrtBA permease controls heme toxicity by its direct and specific efflux. The use of an intracellular heme sensor to control heme efflux constitutes a novel paradigm for bacterial heme homeostasis. PMID:22084241

  20. Discovery of intracellular heme-binding protein HrtR, which controls heme efflux by the conserved HrtB-HrtA transporter in Lactococcus lactis.

    Science.gov (United States)

    Lechardeur, Delphine; Cesselin, Bénédicte; Liebl, Ursula; Vos, Marten H; Fernandez, Annabelle; Brun, Célia; Gruss, Alexandra; Gaudu, Philippe

    2012-02-10

    Most commensal and food bacteria lack heme biosynthesis genes. For several of these, the capture of environmental heme is a means of activating aerobic respiration metabolism. Our previous studies in the Gram-positive bacterium Lactococcus lactis showed that heme exposure strongly induced expression of a single operon, called here hrtRBA, encoding an ortholog of the conserved membrane hrt (heme-regulated transporter) and a unique transcriptional regulator that we named HrtR. We show that HrtR expressed as a fusion protein is a heme-binding protein. Heme iron interaction with HrtR is non-covalent, hexacoordinated, and involves two histidines, His-72 and His-149. HrtR specifically binds a 15-nt palindromic sequence in the hrtRBA promoter region, which is needed for hrtRBA repression. HrtR-DNA binding is abolished by heme addition, which activates expression of the HrtB-HrtA (HrtBA) transporter in vitro and in vivo. The use of HrtR as an intracellular heme sensor appears to be conserved among numerous commensal bacteria, in contrast with numerous Gram-positive pathogens that use an extracellular heme-sensing system, HssRS, to regulate hrt. Finally, we show for the first time that HrtBA permease controls heme toxicity by its direct and specific efflux. The use of an intracellular heme sensor to control heme efflux constitutes a novel paradigm for bacterial heme homeostasis.

  1. Bruxism and prosthetic treatment: a critical review.

    Science.gov (United States)

    Johansson, Anders; Omar, Ridwaan; Carlsson, Gunnar E

    2011-07-01

    Based on the findings from available research on bruxism and prosthetic treatment published in the dental literature, an attempt was made to draw conclusions about the existence of a possible relationship between the two, and its clinical relevance. MEDLINE/PubMed searches were conducted using the terms 'bruxism' and 'prosthetic treatment', as well as combinations of these and related terms. The few studies judged to be relevant were critically reviewed, in addition to papers found during an additional manual search of reference lists within selected articles. Bruxism is a common parafunctional habit, occurring both during sleep and wakefulness. Usually it causes few serious effects, but can do so in some patients. The etiology is multifactorial. There is no known treatment to stop bruxism, including prosthetic treatment. The role of bruxism in the process of tooth wear is unclear, but it is not considered a major cause. As informed by the present critical review, the relationship between bruxism and prosthetic treatment is one that relates mainly to the effect of the former on the latter. Bruxism may be included among the risk factors, and is associated with increased mechanical and/or technical complications in prosthodontic rehabilitation, although it seems not to affect implant survival. When prosthetic intervention is indicated in a patient with bruxism, efforts should be made to reduce the effects of likely heavy occlusal loading on all the components that contribute to prosthetic structural integrity. Failure to do so may indicate earlier failure than is the norm. Copyright © 2011 Japan Prosthodontic Society. Published by Elsevier Ltd. All rights reserved.

  2. YC-1 activation of human soluble guanylyl cyclase has both heme-dependent and heme-independent components

    Science.gov (United States)

    Martin, E.; Lee, Y. C.; Murad, F.

    2001-01-01

    YC-1 [3-(5'-hydroxymethyl-2'furyl)-1-benzyl indazole] is an allosteric activator of soluble guanylyl cyclase (sGC). YC-1 increases the catalytic rate of the enzyme and sensitizes the enzyme toward its gaseous activators nitric oxide or carbon monoxide. In other studies the administration of YC-1 to experimental animals resulted in the inhibition of the platelet-rich thrombosis and a decrease of the mean arterial pressure, which correlated with increased cGMP levels. However, details of YC-1 interaction with sGC and enzyme activation are incomplete. Although evidence in the literature indicates that YC-1 activation of sGC is strictly heme-dependent, this report presents evidence for both heme-dependent and heme-independent activation of sGC by YC-1. The oxidation of the sGC heme by 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one completely inhibited the response to NO, but only partially attenuated activation by YC-1. We also observed activation by YC-1 of a mutant sGC, which lacks heme. These findings indicate that YC-1 activation of sGC can occur independently of heme, but that activation is substantially increased when the heme moiety is present in the enzyme.

  3. Optimising the prescription of prosthetic technologies (opptec): Outcome measures for evidence based prosthetic practice and use

    LENUS (Irish Health Repository)

    Ryall, Dr Nicola

    2010-01-01

    This study provided a forum for patients and service providers to voice their opinions in what they believe to be the important predictors and outcomes involved in successful rehabilitation following limb loss. To develop a consensus on the most important outcomes and factors to address for both the lower limb and upper limb prosthetic prescription process, the above data relating to lower limb and upper prosthetics were subsequently used in the next phase of the research involving two Delphi surveys of 23 and 53 experts within the lower limb and upper limb amputation and prosthetic field respectively, including users, service providers and researchers.\\r\

  4. Mesofluidic controlled robotic or prosthetic finger

    Science.gov (United States)

    Lind, Randall F; Jansen, John F; Love, Lonnie J

    2013-11-19

    A mesofluidic powered robotic and/or prosthetic finger joint includes a first finger section having at least one mesofluidic actuator in fluid communication with a first actuator, a second mesofluidic actuator in fluid communication with a second actuator and a second prosthetic finger section pivotally connected to the first finger section by a joint pivot, wherein the first actuator pivotally cooperates with the second finger to provide a first mechanical advantage relative to the joint point and wherein the second actuator pivotally cooperates with the second finger section to provide a second mechanical advantage relative to the joint point.

  5. [Prosthetic rehabilitation: needs in Senegalese dental offices].

    Science.gov (United States)

    Mbodj, E B; Diouf, M; Faye, D; Ndiaye, A; Seck, M T; Ndiaye, C; Diallo, P D

    2011-12-01

    Knowledge of dental prosthetic needs will develop strategies for prevention and treatment through a package of individual, community and professional policies. The aim of this study was to evaluate prosthetic needs in Senegalese dental offices. The survey was conducted among people aged 15 years and more attending Senegalese dental clinics. The mean number of missing teeth was 4.4. Only 55.3% of the sample expressed the need for dentures and 81.8% had a diagnosed need for prosthesis. A statistically significant difference was noticed between the needs diagnosed and the expressed needs (p dental offices.

  6. Implication for using heme methyl hyperfine shifts as indicators of heme seating as related to stereoselectivity in the catabolism of heme by heme oxygenase: in-plane heme versus axial his rotation.

    Science.gov (United States)

    Ogura, Hiroshi; Evans, John P; de Montellano, Paul R Ortiz; La Mar, Gerd N

    2008-01-08

    The triple mutant of the solubilized, 265-residue construct of human heme oxygenase, K18E/E29K/R183E-hHO, has been shown to redirect the exclusive alpha-regioselectivity of wild-type hHO to primarily beta,delta-selectivity in the cleavage of heme (Wang, J., Evans, J. P., Ogura, H., La Mar, G. N., and Ortiz de Montellano, P. R. (2006) Biochemistry 45, 61-73). The 1H NMR hyperfine shift pattern for the substrate and axial His CbetaH's and the substrate-protein contacts of the cyanide-inhibited protohemin and 2,4-dimethyldeuterohemin complexes of the triple mutant have been analyzed in detail and compared to data for the WT complex. It is shown that protein contacts for the major solution isomers for both substrates in the mutant dictate approximately 90 degrees in-plane clockwise rotation relative to that in the WT. The conventional interpretation of the pattern of substrate methyl hyperfine shifts, however, indicates substrate rotations of only approximately 50 degrees . This paradox is resolved by demonstrating that the axial His25 imidazole ring also rotates counterclockwise with respect to the protein matrix in the mutant relative to that in the WT. The axial His25 CbetaH hyperfine shifts are shown to serve as independent probes of the imidazole plane orientation relative to the protein matrix. The analysis indicates that the pattern of heme methyl hyperfine shifts cannot be used alone to determine the in-plane orientation of the substrate as it relates to the stereospecificity of heme cleavage, without explicit consideration of the orientation of the axial His imidazole plane relative to the protein matrix.

  7. Impairment of heme synthesis in myelin as potential trigger of multiple sclerosis.

    Science.gov (United States)

    Morelli, Alessandro; Ravera, Silvia; Calzia, Daniela; Panfoli, Isabella

    2012-06-01

    The pathogenesis of multiple sclerosis (MS), a disease characterized by demyelination and subsequent axonal degeneration, is as yet unknown. Also, the nature of the disease is as yet not established, since doubts have been cast on its autoimmune origin. Genetic and environmental factors have been implied in MS, leading to the idea of an overall multifactorial origin. An unexpected role in energizing the axon has been reported for myelin, supposed to be the site of consumption of most of oxygen in brain. Myelin would be able to perform oxidative phosphorylation to supply the axons with ATP, thanks to the expression therein of mitochondrial F(o)F(1)-ATP synthase, and respiratory chains. Interestingly, myelin expresses the pathway of heme synthesis, hence of cytochromes, that rely on heme group, in turn depending on Fe availability. Poisoning by these pollutants shares the common characteristic to bring about demyelination both in animal models and in man. Carbon monoxide (CO) and lead poisoning which cause functional imbalance of the heme group, as well as of heme synthesis, cause myelin damage. On the other hand, a lack of essential metals such as iron and copper, produces dramatic myelin decrease. Myelin is a primary target, of iron shortage, indicating that in myelin Fe-dependent processes are more active than in other tissues. The predominant spread of MS in industrialized countries where pollution by heavy metals, and CO poisoning is widespread, suggests a relationship among toxic action of metal pollutants and MS. According to the present hypothesis, MS can be primarily triggered by environmental factors acting on a genetic susceptibility, while the immune response may be a consequence of a primary oxidative damage due to reactive oxygen species produced consequently to an imbalance of cytochromes and respiratory chains in the sheath. Copyright © 2012 Elsevier Ltd. All rights reserved.

  8. Spectroscopic characterization of mononitrosyl complexes in heme-nonheme diiron centers within the myoglobin scaffold (FeBMbs): relevance to denitrifying NO reductase†

    Science.gov (United States)

    Hayashi, Takahiro; Miner, Kyle D.; Yeung, Natasha; Lin, Ying-Wu; Lu, Yi; Moënne-Loccoz, Pierre

    2011-01-01

    Denitrifying NO reductases are evolutionarily related to the superfamily of heme-copper terminal oxidases. These transmembrane protein complexes utilize a heme-nonheme diiron center to reduce two NO molecules to N2O. To understand this reaction, the diiron site has been modeled using sperm whale myoglobin as a scaffold and mutating distal residues Leu-29 and Phe-43 to histidines, and Val-68 to a glutamic acid to create a nonheme FeB site. The impact of incorporation of metal ions at this engineered site on the reaction of the ferrous heme with one NO was examined by UV-vis absorption, EPR, resonance Raman, and FTIR spectroscopies. UV-vis absorption and resonance Raman spectra demonstrate that the first NO molecule binds to the ferrous heme, but while the apoproteins and CuI- or ZnII-loaded proteins show characteristic EPR signatures of S = 1/2 six-coordinate heme {FeNO}7 species observable at liquid nitrogen temperature, the FeII-loaded proteins are EPR silent at ≥ 30 K. Vibrational modes from the heme [Fe-N-O] unit are identified in the RR and FTIR spectra using 15NO and 15N18O. The apo- and CuI-bound proteins exhibit ν(FeNO) and ν(NO) that are only marginally distinct from those reported for native myoglobin. However, binding of FeII at the FeB site shifts the heme ν(FeNO) by +17 cm-1 and the ν(NO) by -50 cm-1 to 1549 cm-1. This low ν(NO) is without precedent for a six-coordinate heme {FeNO}7 species and suggests that the NO group adopts a strong nitroxyl character stabilized by electrostatic interaction with the nearby nonheme FeII. Detection of a similarly low ν(NO) in the ZnII-loaded protein supports this interpretation. PMID:21634416

  9. 21 CFR 895.101 - Prosthetic hair fibers.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Prosthetic hair fibers. 895.101 Section 895.101...) MEDICAL DEVICES BANNED DEVICES Listing of Banned Devices § 895.101 Prosthetic hair fibers. Prosthetic hair fibers are devices intended for implantation into the human scalp to simulate natural hair or conceal...

  10. Comparative roll-over analysis of prosthetic feet

    NARCIS (Netherlands)

    Curtze, Carolin; Hof, At L.; van Keeken, Helco G.; Halbertsma, Jan P. K.; Postema, Klaas; Otten, Bert

    2009-01-01

    A prosthetic foot is a key element of a prosthetic leg, literally forming the basis for a stable and efficient amputee gait. We determined the roll-over characteristics of a broad range of prosthetic feet and examined the effect of a variety of shoes on these characteristics. The body weight of a

  11. Staphylococcus capitis isolated from prosthetic joint infections.

    Science.gov (United States)

    Tevell, S; Hellmark, B; Nilsdotter-Augustinsson, Å; Söderquist, B

    2017-01-01

    Further knowledge about the clinical and microbiological characteristics of prosthetic joint infections (PJIs) caused by different coagulase-negative staphylococci (CoNS) may facilitate interpretation of microbiological findings and improve treatment algorithms. Staphylococcus capitis is a CoNS with documented potential for both human disease and nosocomial spread. As data on orthopaedic infections are scarce, our aim was to describe the clinical and microbiological characteristics of PJIs caused by S. capitis. This retrospective cohort study included three centres and 21 patients with significant growth of S. capitis during revision surgery for PJI between 2005 and 2014. Clinical data were extracted and further microbiological characterisation of the S. capitis isolates was performed. Multidrug-resistant (≥3 antibiotic groups) S. capitis was detected in 28.6 % of isolates, methicillin resistance in 38.1 % and fluoroquinolone resistance in 14.3 %; no isolates were rifampin-resistant. Heterogeneous glycopeptide-intermediate resistance was detected in 38.1 %. Biofilm-forming ability was common. All episodes were either early post-interventional or chronic, and there were no haematogenous infections. Ten patients experienced monomicrobial infections. Among patients available for evaluation, 86 % of chronic infections and 70 % of early post-interventional infections achieved clinical cure; 90 % of monomicrobial infections remained infection-free. Genetic fingerprinting with repetitive sequence-based polymerase chain reaction (rep-PCR; DiversiLab®) displayed clustering of isolates, suggesting that nosocomial spread might be present. Staphylococcus capitis has the potential to cause PJIs, with infection most likely being contracted during surgery or in the early postoperative period. As S. capitis might be an emerging nosocomial pathogen, surveillance of the prevalence of PJIs caused by S. capitis could be recommended.

  12. Graduates’ perceptions of prosthetic and orthotic education and clinical practice in Tanzania and Malawi

    Directory of Open Access Journals (Sweden)

    Lina Magnusson

    2016-02-01

    Full Text Available Background: Maintaining and improving the quality of prosthetics and orthotics education at the Tanzania Training Centre for Orthopaedic Technologists is essential for the provision of appropriate prosthetics and orthotics services in African countries.Objectives: To describe how Tanzanian and Malawian graduates’ of the Diploma in Orthopaedic Technology perceive their education and how it could be improved or supplemented to facilitate clinical practice of graduates.Methods: Nineteen graduates from the diploma course in orthopaedic technology were interviewed and phenomenographic analysis was applied to the data.Results: Seven descriptive categories emerged, namely varied awareness of the profession before starting education, well-equipped teaching facilities, aspects lacking in the learning context, need for changes in the curriculum, enabling people to walk is motivating, obstacles in working conditions and the need for continuous professional development. All participants perceived possible improvements to the content and learning environment.Conclusions: Prosthetic and orthotic education can be better provided by modifying the content of the diploma programme by dedicating more time to the clinical management of different patient groups and applied biomechanics as well as reducing the programme content focusing on technical aspects of prosthetic and orthotic practice. Graduates were not prepared for the rural working conditions and the graduates desired continued training.Keywords: orthotic; prosthetic; education; Malawi; Tanzania; assistive device; assistive technology; developing countries; low-income country

  13. Immunolocalization of heme oxygenase-1 in periodontal diseases

    Directory of Open Access Journals (Sweden)

    G Gayathri

    2014-01-01

    Conclusion: The results of our study is an increasing evidence of involvement of antioxidant enzymes like heme oxygenase-1 in periodontal inflammation and their implication for treatment of chronic periodontitis.

  14. Heme Deficiency in Alzheimer's Disease: A Possible Connection to Porphyria

    Science.gov (United States)

    Dwyer, Barney E.; Stone, Meghan L.; Zhu, Xiongwei; Perry, George; Smith, Mark A.

    2006-01-01

    Mechanisms that cause Alzheimer's disease (AD), an invariably fatal neurodegenerative disease, are unknown. Important recent data indicate that neuronal heme deficiency may contribute to AD pathogenesis. If true, factors that contribute to the intracellular heme deficiency could potentially alter the course of AD. The porphyrias are metabolic disorders characterized by enzyme deficiencies in the heme biosynthetic pathway. We hypothesize that AD may differ significantly in individuals possessing the genetic trait for an acute hepatic porphyria. We elaborate on this hypothesis and briefly review the characteristics of the acute hepatic porphyrias that may be relevant to AD. We note the proximity of genes encoding enzymes of the heme biosynthesis pathway to genetic loci linked to sporadic, late-onset AD. In addition, we suggest that identification of individuals carrying the genetic trait for acute porphyria may provide a unique resource for investigating AD pathogenesis and inform treatment and management decisions. PMID:17047301

  15. Gas-phase spectroscopy of ferric heme-NO complexes

    DEFF Research Database (Denmark)

    Wyer, J.A.; Jørgensen, Anders; Pedersen, Bjarke

    2013-01-01

    and significantly blue-shifted compared to ferric heme nitrosyl proteins (maxima between 408 and 422 nm). This is in stark contrast to the Q-band absorption where the protein microenvironment is nearly innocent in perturbing the electronic structure of the porphyrin macrocycle. Photodissociation is primarily...... through loss of NO. In contrast to the Q-band region, two-photon absorption was seen in the Soret band despite NO loss only requiring ∼1 eV. A model based on intersystem crossing to a long-lived triplet state where a barrier has to be surmounted is suggested. Finally, we summarise the measured absorption...... maxima of heme and its complexes with amino acids and NO. Not so innocent: Weakly bound complexes between ferric heme and NO were synthesised in the gas phase, and their absorption measured from photodissociation yields. Opposite absorption trends in the Soret-band are seen upon NO addition to heme ions...

  16. FORUM: Bioinspired Heme, Heme/non-heme Diiron, Heme/copper and Inorganic NOx Chemistry: ·NO(g) Oxidation, Peroxynitrite-Metal Chemistry and ·NO(g) Reductive Coupling

    Science.gov (United States)

    Schopfer, Mark P.; Wang, Jun; Karlin, Kenneth D.

    2010-01-01

    The focus of this Forum review highlights work from our own laboratories and those of others in the area of biochemical and biologically inspired inorganic chemistry dealing with nitric oxide (nitrogen monoxide, ·NO(g)) and its biological roles and reactions. The latter focus is on (i) oxidation of ·NO(g) to nitrate by nitric oxide dioxygenases (NOD’s), and (ii) reductive coupling of two molecules of ·NO(g) to give N2O(g). In the former case, NOD’s are described and the highlighting of possible peroxynitrite-heme intermediates and consequences of this are given by discussion of recent works with myoglobin and a synthetic heme model system for NOD action. Summaries of recent copper complex chemistries with ·NO(g) and O2(g) leading to peroxynitrite species are given. The coverage of biological reductive coupling of ·NO(g) deals with bacterial nitric oxide reductases (NOR’s) with heme/non-heme diiron active sites, and on heme/Cu oxidases such as cytochrome c oxidase which can mediate the same chemistry. Recent designed protein and synthetic model compound (heme/non-heme diiron or heme/copper) as functional mimics are discussed in some detail. We also highlight examples from the chemical literature, not necessarily involving biologically relevant metal ions, which describe the oxidation of ·NO(g) to nitrate (or nitrite) and possible peroxynitrite intermediates, or reductive coupling of ·NO(g) to give nitrous oxide. PMID:20666386

  17. Isocyanides inhibit human heme oxygenases at the verdoheme stage.

    Science.gov (United States)

    Evans, John P; Kandel, Sylvie; Ortiz de Montellano, Paul R

    2009-09-22

    Heme oxygenases (HO) catalyze the oxidative cleavage of heme to generate biliverdin, CO, and free iron. In humans, heme oxygenase-1 (hHO-1) is overexpressed in tumor tissues, where it helps to protect cancer cells from anticancer agents, while HOs in fungal pathogens, such as Candida albicans, function as the primary means of iron acquisition. Thus, HO can be considered a potential therapeutic target for certain diseases. In this study, we have examined the equilibrium binding of three isocyanides, isopropyl, n-butyl, and benzyl, to the two major human HO isoforms (hHO-1 and hHO-2), Candida albicans HO (CaHmx1), and human cytochrome P450 CYP3A4 using electronic absorption spectroscopy. Isocyanides coordinate to both ferric and ferrous HO-bound heme, with tighter binding by the more hydrophobic isocyanides and 200-300-fold tighter binding to the ferrous form. Benzyl isocyanide was the strongest ligand to ferrous heme in all the enzymes. Because the dissociation constants (KD) of the ligands for ferrous heme-hHO-1 were below the limit of accuracy for equilibrium titrations, stopped-flow kinetic experiments were used to measure the binding parameters of the isocyanides to ferrous hHO-1. Steady-state activity assays showed that benzyl isocyanide was the most potent uncompetitive inhibitor with respect to heme with a KI = 0.15 microM for hHO-1. Importantly, single turnover assays revealed that the reaction was completely stopped by coordination of the isocyanide to the verdoheme intermediate rather than to the ferric heme complex. Much tighter binding of the inhibitor to the verdoheme intermediate differentiates it from inhibition of, for example, CYP3A4 and offers a possible route to more selective inhibitor design.

  18. Isocyanides Inhibit Human Heme Oxygenases at the Verdoheme Stage†

    Science.gov (United States)

    Evans, John P.; Kandel, Sylvie; Ortiz de Montellano, Paul R.

    2010-01-01

    Heme oxygenases (HO) catalyze the oxidative cleavage of heme to generate biliverdin, CO, and free iron. In humans, heme oxygenase-1 (hHO-1) is overexpressed in tumor tissues, where it helps to protect cancer cells from anticancer agents, while HOs in fungal pathogens, such as Candida albicans, function as the primary means of iron acquisition. Thus, HO can be considered a potential therapeutic target for certain diseases. In this study, we have examined the equilibrium binding of three isocyanides; isopropyl, n-butyl, and benzyl, to the two major human HO isoforms (hHO-1 and hHO-2), Candida albicans HO (CaHmx1), and human cytochrome P450 CYP3A4 using electronic absorption spectroscopy. Isocyanides coordinate to both ferric and ferrous HO-bound heme, with tighter binding by the more hydrophobic isocyanides, and 200-300-fold tighter binding to the ferrous form. Benzyl isocyanide was the strongest ligand to ferrous heme in all the enzymes. Because the dissociation constants (KD) of the ligands for ferrous heme-hHO-1 were below the limit of accuracy for equilibrium titrations, stopped-flow kinetic experiments were used to measure the binding parameters of the isocyanides to ferrous hHO-1. Steady-state activity assays showed that benzyl isocyanide was the most potent uncompetitive inhibitor with respect to heme with a KI = 0.15 μM for hHO-1. Importantly, single turnover assays revealed that the reaction was completely stopped by coordination of the isocyanide to the verdoheme intermediate rather than to the ferric heme complex. Much tighter binding of the inhibitor to the verdoheme intermediate differentiates it from inhibition of, for example, CYP3A4 and offers a possible route to more selective inhibitor design. PMID:19694439

  19. Genome-wide analysis reveals novel genes essential for heme homeostasis in Caenorhabditis elegans.

    Directory of Open Access Journals (Sweden)

    Scott Severance

    2010-07-01

    Full Text Available Heme is a cofactor in proteins that function in almost all sub-cellular compartments and in many diverse biological processes. Heme is produced by a conserved biosynthetic pathway that is highly regulated to prevent the accumulation of heme--a cytotoxic, hydrophobic tetrapyrrole. Caenorhabditis elegans and related parasitic nematodes do not synthesize heme, but instead require environmental heme to grow and develop. Heme homeostasis in these auxotrophs is, therefore, regulated in accordance with available dietary heme. We have capitalized on this auxotrophy in C. elegans to study gene expression changes associated with precisely controlled dietary heme concentrations. RNA was isolated from cultures containing 4, 20, or 500 microM heme; derived cDNA probes were hybridized to Affymetrix C. elegans expression arrays. We identified 288 heme-responsive genes (hrgs that were differentially expressed under these conditions. Of these genes, 42% had putative homologs in humans, while genomes of medically relevant heme auxotrophs revealed homologs for 12% in both Trypanosoma and Leishmania and 24% in parasitic nematodes. Depletion of each of the 288 hrgs by RNA-mediated interference (RNAi in a transgenic heme-sensor worm strain identified six genes that regulated heme homeostasis. In addition, seven membrane-spanning transporters involved in heme uptake were identified by RNAi knockdown studies using a toxic heme analog. Comparison of genes that were positive in both of the RNAi screens resulted in the identification of three genes in common that were vital for organismal heme homeostasis in C. elegans. Collectively, our results provide a catalog of genes that are essential for metazoan heme homeostasis and demonstrate the power of C. elegans as a genetic animal model to dissect the regulatory circuits which mediate heme trafficking in both vertebrate hosts and their parasites, which depend on environmental heme for survival.

  20. Consumer satisfaction in prosthetics and orthotics facilities

    NARCIS (Netherlands)

    Geertzen, J.H.B.; Gankema, H.G.J.; Groothoff, J.W.; Dijkstra, P.U.

    The aim of this study was to assess consumer/patient satisfaction with the services of the prosthetics and orthotics (P&O) facilities in the north of the Netherlands, using a modified SERVQUAL questionnaire. In this questionnaire, consumer interests and experiences are assessed on a 5-point Likert

  1. Successful Thrombolysis of Aortic Prosthetic Valve Thrombosis ...

    African Journals Online (AJOL)

    Arun Kumar Agnihotri

    She delivered a normal baby uneventfully in follow up at full term of pregnancy with no complications. Fibrinolytic therapy for mechanical valve thrombosis is a reasonable alternative to surgery in first trimester of pregnancy. KEY WORDS: Prosthetic valve thrombosis; Echocardiography; Streptokinase;. Thrombolysis; Fetus.

  2. Prosthetic Management of Patients Presenting with Juvenile ...

    African Journals Online (AJOL)

    Conclusion: The main objective of prosthetic rehabilitation is the restoration of the integrity of the dental arches. However, the choice of teeth replaced in the partial dentures provided for the patients in this study was based on the choice of the patients, and their choice was determined by aesthetics and affordability.

  3. Promoting prosthetics in the Latino community.

    Science.gov (United States)

    Jaramillo, R; Barabe, J G; Cupp, D

    1995-01-01

    This article describes an effective approach to informing the Latino community about prosthetics. Unlike English, little information on this subject is available in Spanish. The process of obtaining, fabricating, and wearing a prosthesis was interwoven into the teleplay "Milagros." The story concept, video production, and the Latino population's cultural characteristics are discussed. The audience welcomed the opportunity to share the information with others.

  4. Chromosomal localization of the human heme oxygenase genes: Heme oxygenase-1 (HMOX1) maps to chromosome 22q12 and heme oxygenase-2 (HMOX2) maps to chromosome 16p13. 3

    Energy Technology Data Exchange (ETDEWEB)

    Kutty, R.K.; Kutty, G.; Rodriguez, I.R.; Chader, G.J.; Wiggert, B. (National Institutes of Health, Bethesda, MD (United States))

    1994-04-01

    Heme oxygenase catalyzes the oxidation of heme to biliverdin, the precursor of the bile pigment bilirubin, and carbon monoxide, a putative neurotransmitter. The authors have employed polymerase chain reaction and fluorescence in situ hybridization to determine the chromosome localization of the genes coding for the two known heme oxygenase isozymes. Heme oxygenase-1 (HMOX1), the inducible form, was localized to human chromosome 22q12, while heme oxygenase-2 (HMOX2), the constitutive form, was localized to chromosome 16p13.3. 14 refs., 3 figs.

  5. Global changes in Staphylococcus aureus gene expression during human prosthetic joint infection

    DEFF Research Database (Denmark)

    Xu, Yijuan; Nielsen, Per Halkjær; Nielsen, Jeppe Lund

    2016-01-01

    Global changes in Staphylococcus aureus gene expression during human prosthetic joint infection Xu, Yijuan1; Nielsen, Per H.1; Nielsen, Jeppe L.1; Thomsen, Trine R. 1,2; Nielsen, Kåre L.1 and the PRIS study group 1: Center for Microbial Communities, Department of Biotechnology, Chemistry...... and Environmental Engineering, Aalborg University, Denmark 2: Danish Technological Institute, Aarhus, Denmark Aim: ”The aim of this study was to gain insight into the in vivo expression of virulence and metabolic genes of Staphylococcus aureus in a prosthetic joint infection in a human subject” Method: ”Deep RNA...... sequencing (RNA-seq) was used for transcriptome profile of joint fluid obtained from a patient undergoing surgery due to acute S. aureus prosthetic joint infection. The S. aureus gene expression in the infection was compared with exponential culture of a S. aureus isolate obtained from the same sample using...

  6. Acquisition of iron from transferrin regulates reticulocyte heme synthesis

    Energy Technology Data Exchange (ETDEWEB)

    Ponka, P.; Schulman, H.M.

    1985-11-25

    Fe-salicylaldehyde isonicotinoylhydrazone (SIH), which can donate iron to reticulocytes without transferrin as a mediator, has been utilized to test the hypothesis that the rate of iron uptake from transferrin limits the rate of heme synthesis in erythroid cells. Reticulocytes take up VZFe from (VZFe)SIH and incorporate it into heme to a much greater extent than from saturating concentrations of (VZFe)transferrin. Also, Fe-SIH stimulates (2- UC)glycine into heme when compared to the incorporation observed with saturating levels of Fe-transferrin. In addition, delta-aminolevulinic acid does not stimulate VZFe incorporation into heme from either (VZFe)transferrin or (VZFe)SIH but does reverse the inhibition of VZFe incorporation into heme caused by isoniazid, an inhibitor of delta-aminolevulinic acid synthase. Taken together, these results suggest the hypothesis that some step(s) in the pathway of iron from extracellular transferrin to intracellular protoporphyrin limits the overall rate of heme synthesis in reticulocytes.

  7. A Comparative Study of O2, CO and CN Binding to Heme IX Protein Models

    Directory of Open Access Journals (Sweden)

    Francisco Torrens

    2004-07-01

    Full Text Available Parametrization of a molecular-mechanics program to include terms specificfor five- and six-coordinate transition metal complexes results in computer-simulatedstructures of heme complexes. The principal new feature peculiar to five and sixcoordination is a term that measures the effect of electron-pair repulsion modified by theligand electronegativity and takes into account the different structural possibilities. Themodel system takes into account the structural differences of the fixing centre in thehaemoglobin subunits. The customary proximal histidine is added. The prosthetic groupheme IX is wholly considered in our model. The calculations show clearly that certainconformations are much more favourable that others for fixing O2. From the O2 bindingin haemoglobin, myoglobin and simple Fe porphyrin models it is concluded that the bentO2 ligand is best viewed as bound superoxide O2–. Axial ligands are practically free-rotating. A small modification of the model in both crystal and protein matrix affects theorientation of the ligands in experimental systems.

  8. Pregnancy after Prosthetic Aortic Valve Replacement: How Do We Monitor Prosthetic Valvular Function during Pregnancy?

    Directory of Open Access Journals (Sweden)

    Nicole Sahasrabudhe

    2018-01-01

    Full Text Available Background. With modern medicine, many women after structural heart repair are deciding to experience pregnancy. There is a need for further study to identify normal echocardiographic parameters to better assess prosthetic valvular function in pregnancy. In addition, a multidisciplinary approach is essential in managing pregnant patients with complex cardiac conditions. Case. A 22-year-old nulliparous woman with an aortic valve replacement 18 months prior to her pregnancy presented to prenatal care at 20-week gestation. During her prenatal care, serial echocardiography showed a significant increase in the mean gradient across the prosthetic aortic valve. Multidisciplinary management and a serial echocardiography played an integral role in her care that resulted in a successful spontaneous vaginal delivery without complications. Conclusion. Further characterization of the normal echocardiographic parameters in pregnant patients with prosthetic valves is critical to optimize prenatal care for this patient population. This case report is novel in that serial echocardiograms were obtained throughout prenatal care, which showed significant changes across the prosthetic aortic valve. Teaching Points. (1 Further study is needed to identify normal echocardiographic parameters to best assess prosthetic valvular function in pregnancy. (2 Multidisciplinary management is encouraged to optimize prenatal care for women with prosthetic aortic valve replacements.

  9. Do pH and flavonoids influence hypochlorous acid-induced catalase inhibition and heme modification?

    Science.gov (United States)

    Krych-Madej, Justyna; Gebicka, Lidia

    2015-09-01

    Hypochlorous acid (HOCl), highly reactive oxidizing and chlorinating species, is formed in the immune response to invading pathogens by the reaction of hydrogen peroxide with chloride catalyzed by the enzyme myeloperoxidase. Catalase, an important antioxidant enzyme, catalyzing decomposition of hydrogen peroxide to water and molecular oxygen, hampers in vitro HOCl formation, but is also one of the main targets for HOCl. In this work we have investigated HOCl-induced catalase inhibition at different pH, and the influence of flavonoids (catechin, epigallocatechin gallate and quercetin) on this process. It has been shown that HOCl-induced catalase inhibition is independent on pH in the range 6.0-7.4. Preincubation of catalase with epigallocatechin gallate and quercetin before HOCl treatment enhances the degree of catalase inhibition, whereas catechin does not affect this process. Our rapid kinetic measurements of absorption changes around the heme group have revealed that heme modification by HOCl is mainly due to secondary, intramolecular processes. The presence of flavonoids, which reduce active catalase intermediate, Compound I to inactive Compound II have not influenced the kinetics of HOCl-induced heme modification. Possible mechanisms of the reaction of hypochlorous acid with catalase are proposed and the biological consequences are discussed. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Expression and activity analysis reveal that heme oxygenase (decycling) 1 is associated with blue egg formation.

    Science.gov (United States)

    Wang, Z P; Liu, R F; Wang, A R; Li, J Y; Deng, X M

    2011-04-01

    Biliverdin is responsible for the coloration of blue eggs and is secreted onto the eggshell by the shell gland. Previous studies confirmed that a significant difference exists in biliverdin content between blue eggs and brown eggs, although the reasons are still unknown. Because the pigment is derived from oxidative degradation of heme catalyzed by heme oxygenase (HO), this study compared heme oxygenase (decycling) 1 (HMOX1), the gene encoding HO expression and HO activity, in the shell glands of the Dongxiang blue-shelled chicken (n = 12) and the Dongxiang brown-shelled chicken (n = 12). Results showed that HMOX1 was highly expressed at the mRNA (1.58-fold; P 0.05). Taken together, these results show that blue egg formation is associated with high expression of HMOX1 in the shell gland of Dongxiang blue-shelled chickens, and suggest that differential expression of HMOX1 in the 2 groups of chickens is most likely to arise from an alteration in the trans-acting factor.

  11. Comparative roll-over analysis of prosthetic feet.

    Science.gov (United States)

    Curtze, Carolin; Hof, At L; van Keeken, Helco G; Halbertsma, Jan P K; Postema, Klaas; Otten, Bert

    2009-08-07

    A prosthetic foot is a key element of a prosthetic leg, literally forming the basis for a stable and efficient amputee gait. We determined the roll-over characteristics of a broad range of prosthetic feet and examined the effect of a variety of shoes on these characteristics. The body weight of a person acting on a prosthetic foot during roll-over was emulated by means of an inverted pendulum-like apparatus. Parameters measured were the effective radius of curvature, the forward travel of the center of pressure, and the instantaneous radius of curvature of the prosthetic feet. Finally, we discuss how these parameters relate to amputee gait.

  12. HemeBIND: a novel method for heme binding residue prediction by combining structural and sequence information

    Directory of Open Access Journals (Sweden)

    Hu Jianjun

    2011-05-01

    Full Text Available Abstract Background Accurate prediction of binding residues involved in the interactions between proteins and small ligands is one of the major challenges in structural bioinformatics. Heme is an essential and commonly used ligand that plays critical roles in electron transfer, catalysis, signal transduction and gene expression. Although much effort has been devoted to the development of various generic algorithms for ligand binding site prediction over the last decade, no algorithm has been specifically designed to complement experimental techniques for identification of heme binding residues. Consequently, an urgent need is to develop a computational method for recognizing these important residues. Results Here we introduced an efficient algorithm HemeBIND for predicting heme binding residues by integrating structural and sequence information. We systematically investigated the characteristics of binding interfaces based on a non-redundant dataset of heme-protein complexes. It was found that several sequence and structural attributes such as evolutionary conservation, solvent accessibility, depth and protrusion clearly illustrate the differences between heme binding and non-binding residues. These features can then be separately used or combined to build the structure-based classifiers using support vector machine (SVM. The results showed that the information contained in these features is largely complementary and their combination achieved the best performance. To further improve the performance, an attempt has been made to develop a post-processing procedure to reduce the number of false positives. In addition, we built a sequence-based classifier based on SVM and sequence profile as an alternative when only sequence information can be used. Finally, we employed a voting method to combine the outputs of structure-based and sequence-based classifiers, which demonstrated remarkably better performance than the individual classifier alone

  13. Outdoor weathering of facial prosthetic elastomers differing in Durometer hardness.

    Science.gov (United States)

    Willett, Emily S; Beatty, Mark W

    2015-03-01

    Facial prosthetic elastomers with wide ranges in hardness are available, yet material weatherability is unknown. The purpose of this study was to assess color, Durometer hardness, and tensile property changes after 3000 hours of outdoor weathering. Unpigmented elastomers with Durometer hardness 5, 30, 50, 70, and A-2186 were polymerized into dumbbells (ASTM D412) and disks, 34 mm in diameter by 6 mm thick. Materials were subjected to outdoor or time passage environments for 3000 hours. CIELab color (n=5), Durometer hardness (n=5), and tensile mechanical properties (n=10) were measured at 0 and 3000 hours, and group differences were assessed by material and weathering condition (ANOVA/Tukey, α=.05). Except for A-2186, the mean Durometer changes for all materials were 1 unit or less, with no significant differences observed between time passage and weathered groups (P≥.05). Three-thousand-hour tensile mechanical property results demonstrated nonsignificant differences between time passage and weathered materials but significantly changed properties from immediately tested materials (Phardness 5 and 30 and A-2186. With a few exceptions, outdoor weathering produced relatively small changes in color, Durometer hardness, or tensile properties compared with time passage. Copyright © 2015 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

  14. Heme Degradation by Heme Oxygenase Protects Mitochondria but Induces ER Stress via Formed Bilirubin

    Directory of Open Access Journals (Sweden)

    Andrea Müllebner

    2015-04-01

    Full Text Available Heme oxygenase (HO, in conjunction with biliverdin reductase, degrades heme to carbon monoxide, ferrous iron and bilirubin (BR; the latter is a potent antioxidant. The induced isoform HO-1 has evoked intense research interest, especially because it manifests anti-inflammatory and anti-apoptotic effects relieving acute cell stress. The mechanisms by which HO mediates the described effects are not completely clear. However, the degradation of heme, a strong pro-oxidant, and the generation of BR are considered to play key roles. The aim of this study was to determine the effects of BR on vital functions of hepatocytes focusing on mitochondria and the endoplasmic reticulum (ER. The affinity of BR to proteins is a known challenge for its exact quantification. We consider two major consequences of this affinity, namely possible analytical errors in the determination of HO activity, and biological effects of BR due to direct interaction with protein function. In order to overcome analytical bias we applied a polynomial correction accounting for the loss of BR due to its adsorption to proteins. To identify potential intracellular targets of BR we used an in vitro approach involving hepatocytes and isolated mitochondria. After verification that the hepatocytes possess HO activity at a similar level as liver tissue by using our improved post-extraction spectroscopic assay, we elucidated the effects of increased HO activity and the formed BR on mitochondrial function and the ER stress response. Our data show that BR may compromise cellular metabolism and proliferation via induction of ER stress. ER and mitochondria respond differently to elevated levels of BR and HO-activity. Mitochondria are susceptible to hemin, but active HO protects them against hemin-induced toxicity. BR at slightly elevated levels induces a stress response at the ER, resulting in a decreased proliferative and metabolic activity of hepatocytes. However, the proteins that are targeted

  15. Christ Siemens Touraine syndrome: Two case reports and felicitous approaches to prosthetic management.

    Science.gov (United States)

    Renahan, Navanith; Raj, Renju; Varma, R Balagopal; Kumar, J Suresh

    2015-01-01

    Ectodermal dysplasia is a heterogeneous group of inherited disorders, which exhibit a classic triad of hypohydrosis, hypotrichosis, and hypodontia. Hypohidrotic or anhidrotic ectodermal dysplasia or Christ Siemens Touraine syndrome is the most common condition among ectodermal dysplasia patients. This is a case report on two Christ Siemens Touraine syndrome cases and two different approaches to prosthetic management.

  16. Christ Siemens Touraine syndrome: Two case reports and felicitous approaches to prosthetic management

    Directory of Open Access Journals (Sweden)

    Navanith Renahan

    2015-01-01

    Full Text Available Ectodermal dysplasia is a heterogeneous group of inherited disorders, which exhibit a classic triad of hypohydrosis, hypotrichosis, and hypodontia. Hypohidrotic or anhidrotic ectodermal dysplasia or Christ Siemens Touraine syndrome is the most common condition among ectodermal dysplasia patients. This is a case report on two Christ Siemens Touraine syndrome cases and two different approaches to prosthetic management.

  17. Distal anastomotic vein adjunct usage in infrainguinal prosthetic bypasses.

    Science.gov (United States)

    McPhee, James T; Goodney, Philip P; Schanzer, Andres; Shaykevich, Shimon; Belkin, Michael; Menard, Matthew T

    2013-04-01

    Single-segment saphenous vein remains the optimal conduit for infrainguinal revascularization. In its absence, prosthetic conduit may be used. Existing data regarding the significance of anastomotic distal vein adjunct (DVA) usage with prosthetic grafts are based on small series. This is a retrospective cohort analysis derived from the regional Vascular Study Group of New England as well as the Brigham and Women's hospital database. A total of 1018 infrainguinal prosthetic bypass grafts were captured in the dataset from 73 surgeons at 15 participating institutions. Propensity scoring and 3:1 matching was performed to create similar exposure groups for analysis. Outcome measures of interest included: primary patency, freedom from major adverse limb events (MALEs), and amputation free survival at 1 year as a function of vein patch utilization. Time to event data were compared with the log-rank test; multivariable Cox proportional hazard models were used to evaluate the adjusted association between vein cuff usage and the primary end points. DVA was defined as a vein patch, cuff, or boot in any configuration. Of the 1018 bypass operations, 94 (9.2%) had a DVA whereas 924 (90.8%) did not (no DVA). After propensity score matching, 88 DVAs (25%) and 264 no DVAs (75%) were analyzed. On univariate analysis of the matched cohort, the DVA and no DVA groups were similar in terms of mean age (70.0 vs 69.0; P = .55), male sex (58.0% vs 58.3%; P > .99), and preoperative characteristics such as living at home (93.2% vs 94.3%; P = .79) and independent ambulatory status (72.7% vs 75.7%; P = .64). The DVA and no DVA groups had similar rates of major comorbidities such as hypertension chronic obstructive pulmonary disease, diabetes mellitus, coronary artery disease, and dialysis dependence (P > .05 for all). Likewise, they had similar rates of distal origin grafts (13.6% vs 12.5%; P = .85), critical limb ischemia indications (P = .53), and prior arterial bypass (58% vs 47%; P = .08

  18. HEME OXYGENASE-1 AND FATTY LIVER DISEASE.

    Directory of Open Access Journals (Sweden)

    Daniela Nicolosi

    2016-04-01

    Full Text Available Fatty liver diseases are a spectrum of liver pathologies characterized by abnormal hepatocellular accumulations of lipids. This condition may occur in both adults and children, particularly those who are obese or have insulin resistance or following abuse of alcohol consumption. They are classified in Non-Alcoholic Fatty Liver Disease (NAFLD and Alcoholic Fatty Liver Disease (AFLD. Steatohepatitis is a specific pattern of injury within the spectrum of NAFLD and this pattern is associated with fibrotic progression and cirrhosis. The role of oxidative stress in liver steatosis production and its progression to inflammation leading to steatohepatitis has been discussed in relation to alterations in metabolic and pro-inflammatory pathway. One of the main enzymes responsible for antioxidant activity in the presence of liver damage is the Heme Oxygenase-1(HO-1.The products of the HO-1-catalyzed reaction, particularly carbon monoxide (CO and biliverdin/bilirubin have been shown to exert protective effects in several organs against oxidative and other noxious stimuli. In this context, it is interesting to note that induction of HO-1 expression contributes to protection against liver damage in various experimental models. The focus of this review is on the significance of targeted induction of HO-1 as a potential therapeutic strategy to protect the liver against fatty liver diseases.

  19. Dynamics of the His79-heme Alkaline Transition of Yeast Iso-1-cytochrome c Probed by Conformationally-gated Electron Transfer with Co(II)bis(terpyridine)†

    Science.gov (United States)

    Cherney, Melisa M.; Junior, Carolyn C.; Bergquist, Bryan B.; Bowler, Bruce E.

    2013-01-01

    Alkaline conformers of cytochrome c may be involved in both its electron transport and apoptotic functions. We use cobalt(II)bis(terpyridine), Co(terpy)22+, as a reagent for conformationally-gated electron transfer (gated ET) experiments to study the alkaline conformational transition of K79H variants of yeast iso-1-cytochrome c expressed in Escherichia coli, WT*K79H, with alanine at position 72, and Saccharomyces cerevisiae, yK79H, with trimethyllysine (Tml) at position 72. Co(terpy)22+ is well-suited to the 100 ms to 1 s time scale of the His79-mediated alkaline conformational transition of these variants. Reduction of the His79-heme alkaline conformer by Co(terpy)22+ occurs primarily by gated ET, which involves conversion to the native state followed by reduction, with a small fraction of the His79- heme alkaline conformer directly reduced by Co(terpy)22+. The gated ET experiments show that the mechanism of formation of the His79-heme alkaline conformer involves only two ionizable groups. In previous work, we showed that the mechanism of the His73-mediated alkaline conformational transition requires three ionizable groups. Thus, the mechanism of heme crevice opening depends upon the position of the ligand mediating the process. The microscopic rate constants provided by gated ET studies show that mutation of Tml72 (yK79H variant) in the heme crevice loop to Ala72 (WT*K79H variant) affects the dynamics of heme crevice opening through a small destabilization of both the native conformer and the transition state relative to the His79-heme alkaline conformer. Previous pH jump data had indicated that the Tml72→Ala mutation primarily stabilized the transition state for the His79-mediated alkaline conformational transition. PMID:23899348

  20. Human heme oxygenase-1 efficiently catabolizes heme in the absence of biliverdin reductase.

    Science.gov (United States)

    Reed, James R; Huber, Warren J; Backes, Wayne L

    2010-11-01

    Heme oxygenase 1 (HO-1) uses molecular oxygen and electrons from NADPH cytochrome P450 reductase to convert heme to CO, ferrous iron, and biliverdin (BV). Enzymatic studies with the purified 30-kDa form of HO-1 routinely use a coupled assay containing biliverdin reductase (BVR), which converts BV to bilirubin (BR). BVR is believed to be required for optimal HO-1 activity. The goal of this study was to determine whether HO-1 activity could be monitored directly by following BV generation or iron release (using the ferrous iron chelator, ferrozine) in the absence of BVR. Using assays for each of the three end products, we found that HO-1 activity was stimulated in the presence of catalase and comparable rates were measured with each assay. Absorbance scans revealed characteristic spectra for BR, BV, and/or the ferrozine-iron complex. The optimal conditions were slightly different for the direct and coupled assays. BSA activated the coupled but inhibited the direct assays, and the assays had different pH optima. By measuring the activity of BVR directly using BV as a substrate, these differences were attributed to different enzymatic properties of BVR and HO-1. Thus, BVR is not needed to measure the activity of HO-1 when catalase is present. In fact, the factors affecting catalysis by HO-1 are better understood using the direct assays because the coupled assay can be influenced by properties of BVR.

  1. Determination of heme in microorganisms using HPLC-MS/MS and cobalt(III) protoporphyrin IX inhibition of heme acquisition in Escherichia coli.

    Science.gov (United States)

    Fyrestam, Jonas; Östman, Conny

    2017-10-17

    One of the main threats to the achievements in modern medicine is antimicrobial resistance. Molecular targeting of bacterial acquisition mechanisms of heme has been suggested to be an alternative to antibiotics. In the present study, HPLC-MS/MS combined with a simple clean-up based on liquid-liquid extraction has been developed and evaluated for simultaneous determination of heme and porphyrin heme precursors in microorganisms. Experimental design was used to optimize the extraction parameters, to obtain a method with high recovery, low matrix effects, and high precision. The effects of additives in the culture medium on the biosynthesis of heme were studied using Escherichia coli as a model microorganism. 5-Aminolaevulinic acid and hemin increased the heme concentration in E. coli by a factor of 1.5 and 4.5, respectively. Addition of 5-aminolaevulinic acid bypassed the E. coli negative feedback control of heme biosynthesis, which led to high amounts of intracellular porphyrins. The high heme concentration obtained when hemin was used as a culture additive shows that E. coli has an uptake of heme from its surroundings. In contrast, addition of cobalt protoporphyrin IX to the growth medium reduced the amount of heme in E. coli, demonstrating this compound's ability to mimic real heme and inhibit the heme acquisition mechanisms.

  2. Antichaperone activity and heme degradation effect of methyl tert-butyl ether (MTBE) on normal and diabetic hemoglobins.

    Science.gov (United States)

    Najdegerami, Ismaeil Hossein; Maghami, Parvaneh; Sheikh-Hasani, Vahid; Hosseinzadeh, Ghader; Sheibani, Nader; Moosavi-Movahedi, Ali A

    2017-05-01

    Because of the extensive use of methyl tert-butyl ether (MTBE) as an additive to increase the octane quality of gasoline, the environmental pollution by this compound has increased in recent decades. Environmental release of MTBE may lead to its entry to the blood stream through inhalation or drinking of contaminated water, and its interactions with biological molecules such as proteins. The present study was proposed to comparatively investigate the interactions of MTBE with hemoglobin (Hb) from diabetic and nondiabetic individuals using various spectroscopic methods including UV-visible, fluorescence, chemiluminescence, and circular dichroism. These results demonstrated the effects of MTBE on heme degradation of Hb and the reaction of these degradation products with water generating reactive oxygen species. Interaction of Hb with MTBE enhanced its aggregation rate and decreased lag time, indicating the antichaperone activity of MTBE upon interaction with Hb. Furthermore, the diabetic Hb showed more severe effects of MTBE, including heme degradation, reactive oxygen species production, unfolding, and antichaperone behavior than the nondiabetic Hb. The results from molecular docking suggested that the special interaction site of MTBE in the vicinity of Hb heme group is responsible for heme degradation. Copyright © 2016 John Wiley & Sons, Ltd.

  3. [Streptococcus equisimilis associated septic arthritis/prosthetic joint infection].

    Science.gov (United States)

    Sipahi, Oğuz Reşat; Ozkören Calik, Sebnem; Pullukçu, Hüsnü; Işikgöz Taşbakan, Meltem; Arda, Bilgin; Tünger, Alper; Ulusoy, Sercan

    2008-07-01

    Group C streptococci are flora members of skin, nasopharynx, gastrointestinal and genitourinary systems. They are rare causes of human pharyngitis, arthritis, pneumonia, meningitis and bacteremia. In this report, a 71-years old male patient with Streptococcus equisimilis arthritis/prosthetic joint infection has been presented. The patient was admitted to the emergency service with the complaints of erythema, swelling and tenderness on right knee which had total knee prosthesis. Examination of synovial fluid punction sample yielded abundant amount of leukocytes (> 1000 cells/mm3). Empirical ampicillin-sulbactam (1 g q6h, parenterally) therapy was initiated. Bacteria which have been cultivated from synovial fluid specimen were identified as S. equisimilis. The isolate was found to be susceptible to penicilin, erythromycin and teicoplanin, and resistant to chloramphenicol and tetracycline. Although clinical presentation improved during the first ten days, symptoms recurred after the 10th day and the therapy was switched to teicoplanin. The recurrence was thought to be the result of antibiotic tolerence. The patient was treated successfully with teicoplanin, and no relapse or reinfection was observed during one year of follow-up. To our knowledge this is the first case of S. equisimilis arthritis reported from Turkey and first case of S. equisimilis associated prosthetic joint infection.

  4. A surface EMG test tool to measure proportional prosthetic control.

    Science.gov (United States)

    Sturma, Agnes; Roche, Aidan D; Göbel, Peter; Herceg, Malvina; Ge, Nan; Fialka-Moser, Veronika; Aszmann, Oskar

    2015-06-01

    In upper limb amputees, prosthetic control training is recommended before and after fitting. During rehabilitation, the focus is on selective proportional control signals. For functional monitoring, many different tests are available. None can be used in the early phase of training. However, an early assessment is needed to judge if a patient has the potential to control a certain prosthetic set-up. This early analysis will determine if further training is needed or if other strategies would be more appropriate. Presented here is a tool that is capable of predicting achievable function in voluntary EMG control. This tool is applicable to individual muscle groups to support preparation of training and fitting. In four of five patients, the sEMG test tool accurately predicted the suitability for further myoelectric training based on SHAP outcome measures. (P1: "Poor" function in the sEMG test tool corresponded to 54/100 in the SHAP test; P2: Good: 85; P3: Good: 81; P4: Average: 78). One patient scored well during sEMG testing, but was unmotivated during SHAP testing. (Good: 50) Therefore, the surface EMG test tool may predict achievable control skills to a high extent, validated with the SHAP, but requires further clinical testing to validate this technique.

  5. Biophysical Characterisation of Globins and Multi-Heme Cytochromes Using Electron Paramagnetic Resonance and Optical Spectroscopy

    Science.gov (United States)

    Desmet, Filip

    Heme proteins of different families were investigated in this work, using a combination of pulsed and continuous-wave electron paramagnetic resonance (EPR) spectroscopy, optical absorption spectroscopy, resonance Raman spectroscopy and laser flash photolysis. The first class of proteins that were investigated, were the globins. The globin-domain of the globin-coupled sensor of the bacterium Geobacter sulfurreducens was studied in detail using different pulsed EPR techniques (HYSCORE and Mims ENDOR). The results of this pulsed EPR study are compared with the results of the optical investigation and the crystal structure of the protein. The second globin, which was studied, is the Protoglobin of Methanosarcina acetivorans, various mutants of this protein were studied using laser flash photolysis and Raman spectroscopy to unravel the link between this protein's unusual structure and its ligand-binding kinetics. In addition to this, the CN -bound form of this protein was investigated using EPR and the influence of the strong deformation of the heme on the unusual low gz values is discussed. Finally, the neuroglobins of three species of fishes, Danio rerio, Dissostichus mawsoni and Chaenocephalus aceratus are studied. The influence of the presence or absence of two cysteine residues in the C-D and D-region of the protein on the EPR spectrum, and the possible formation of a disulfide bond is studied. The second group of proteins that were studied in this thesis belong to the family of the cytochromes. First the Mouse tumor suppressor cytochrome b561 was studied, the results of a Raman and EPR investigation are compared to the Human orthologue of the protein. Secondly, the tonoplast cytochrome b561 of Arabidopsis was investigated in its natural form and in two double-mutant forms, in which the heme at the extravesicular side was removed. The results of this investigation are then compared with two models in literature that predict the localisation of the hemes in this

  6. Retinal Prosthetics, Optogenetics, and Chemical Photoswitches

    Science.gov (United States)

    2015-01-01

    Three technologies have emerged as therapies to restore light sensing to profoundly blind patients suffering from late-stage retinal degenerations: (1) retinal prosthetics, (2) optogenetics, and (3) chemical photoswitches. Prosthetics are the most mature and the only approach in clinical practice. Prosthetic implants require complex surgical intervention and provide only limited visual resolution but can potentially restore navigational ability to many blind patients. Optogenetics uses viral delivery of type 1 opsin genes from prokaryotes or eukaryote algae to restore light responses in survivor neurons. Targeting and expression remain major problems, but are potentially soluble. Importantly, optogenetics could provide the ultimate in high-resolution vision due to the long persistence of gene expression achieved in animal models. Nevertheless, optogenetics remains challenging to implement in human eyes with large volumes, complex disease progression, and physical barriers to viral penetration. Now, a new generation of photochromic ligands or chemical photoswitches (azobenzene-quaternary ammonium derivatives) can be injected into a degenerated mouse eye and, in minutes to hours, activate light responses in neurons. These photoswitches offer the potential for rapidly and reversibly screening the vision restoration expected in an individual patient. Chemical photoswitch variants that persist in the cell membrane could make them a simple therapy of choice, with resolution and sensitivity equivalent to optogenetics approaches. A major complexity in treating retinal degenerations is retinal remodeling: pathologic network rewiring, molecular reprogramming, and cell death that compromise signaling in the surviving retina. Remodeling forces a choice between upstream and downstream targeting, each engaging different benefits and defects. Prosthetics and optogenetics can be implemented in either mode, but the use of chemical photoswitches is currently limited to downstream

  7. Retinal prosthetics, optogenetics, and chemical photoswitches.

    Science.gov (United States)

    Marc, Robert; Pfeiffer, Rebecca; Jones, Bryan

    2014-10-15

    Three technologies have emerged as therapies to restore light sensing to profoundly blind patients suffering from late-stage retinal degenerations: (1) retinal prosthetics, (2) optogenetics, and (3) chemical photoswitches. Prosthetics are the most mature and the only approach in clinical practice. Prosthetic implants require complex surgical intervention and provide only limited visual resolution but can potentially restore navigational ability to many blind patients. Optogenetics uses viral delivery of type 1 opsin genes from prokaryotes or eukaryote algae to restore light responses in survivor neurons. Targeting and expression remain major problems, but are potentially soluble. Importantly, optogenetics could provide the ultimate in high-resolution vision due to the long persistence of gene expression achieved in animal models. Nevertheless, optogenetics remains challenging to implement in human eyes with large volumes, complex disease progression, and physical barriers to viral penetration. Now, a new generation of photochromic ligands or chemical photoswitches (azobenzene-quaternary ammonium derivatives) can be injected into a degenerated mouse eye and, in minutes to hours, activate light responses in neurons. These photoswitches offer the potential for rapidly and reversibly screening the vision restoration expected in an individual patient. Chemical photoswitch variants that persist in the cell membrane could make them a simple therapy of choice, with resolution and sensitivity equivalent to optogenetics approaches. A major complexity in treating retinal degenerations is retinal remodeling: pathologic network rewiring, molecular reprogramming, and cell death that compromise signaling in the surviving retina. Remodeling forces a choice between upstream and downstream targeting, each engaging different benefits and defects. Prosthetics and optogenetics can be implemented in either mode, but the use of chemical photoswitches is currently limited to downstream

  8. Prosthetic Hand With Two Gripping Fingers

    Science.gov (United States)

    Norton, William E.; Belcher, Jewell B.; Vest, Thomas W.; Carden, James R.

    1993-01-01

    Prosthetic hand developed for amputee who retains significant portion of forearm. Outer end of device is end effector including two fingers, one moved by rotating remaining part of forearm about its longitudinal axis. Main body of end effector is end member supporting fingers, roller bearing assembly, and rack-and-pinion mechanism. Advantage of rack-and-pinion mechanism enables user to open or close gap between fingers with precision and force.

  9. Prosthetic management of an ocular defect

    Directory of Open Access Journals (Sweden)

    Siddesh Kumar Chintal

    2010-01-01

    Full Text Available The disfigurement associated with the loss of an eye can cause significant physical and emotional problems. Various treatment modalities are available, one of which is implants. Although implant has a superior outcome, it may not be advisable in all patients due to economic factors. The present article describes the prosthetic management of an ocular defect with a custom-made ocular prosthesis.

  10. The Prosthetic Workflow in the Digital Era

    OpenAIRE

    Lidia Tordiglione; Michele De Franco; Giovanni Bosetti

    2016-01-01

    The purpose of this retrospective study was to clinically evaluate the benefits of adopting a full digital workflow for the implementation of fixed prosthetic restorations on natural teeth. To evaluate the effectiveness of these protocols, treatment plans were drawn up for 15 patients requiring rehabilitation of one or more natural teeth. All the dental impressions were taken using a Planmeca PlanScan® (Planmeca OY, Helsinki, Finland) intraoral scanner, which provided digital casts on which t...

  11. Heme oxygenase-1 deletion affects stress erythropoiesis.

    Directory of Open Access Journals (Sweden)

    Yu-An Cao

    Full Text Available Homeostatic erythropoiesis leads to the formation of mature red blood cells under non-stress conditions, and the production of new erythrocytes occurs as the need arises. In response to environmental stimuli, such as bone marrow transplantation, myelosuppression, or anemia, erythroid progenitors proliferate rapidly in a process referred to as stress erythropoiesis. We have previously demonstrated that heme oxygenase-1 (HO-1 deficiency leads to disrupted stress hematopoiesis. Here, we describe the specific effects of HO-1 deficiency on stress erythropoiesis.We used a transplant model to induce stress conditions. In irradiated recipients that received hmox(+/- or hmox(+/+ bone marrow cells, we evaluated (i the erythrocyte parameters in the peripheral blood; (ii the staining intensity of CD71-, Ter119-, and CD49d-specific surface markers during erythroblast differentiation; (iii the patterns of histological iron staining; and (iv the number of Mac-1(+-cells expressing TNF-α. In the spleens of mice that received hmox(+/- cells, we show (i decreases in the proerythroblast, basophilic, and polychromatophilic erythroblast populations; (ii increases in the insoluble iron levels and decreases in the soluble iron levels; (iii increased numbers of Mac-1(+-cells expressing TNF-α; and (iv decreased levels of CD49d expression in the basophilic and polychromatophilic erythroblast populations.As reflected by effects on secreted and cell surface proteins, HO-1 deletion likely affects stress erythropoiesis through the retention of erythroblasts in the erythroblastic islands of the spleen. Thus, HO-1 may serve as a therapeutic target for controlling erythropoiesis, and the dysregulation of HO-1 may be a predisposing condition for hematologic diseases.

  12. Adaptive sports technology and biomechanics: prosthetics.

    Science.gov (United States)

    De Luigi, Arthur Jason; Cooper, Rory A

    2014-08-01

    With the technologic advances in medicine and an emphasis on maintaining physical fitness, the population of athletes with impairments is growing. It is incumbent upon health care practitioners to make every effort to inform these individuals of growing and diverse opportunities and to encourage safe exercise and athletic participation through counseling and education. Given the opportunities for participation in sports for persons with a limb deficiency, the demand for new, innovative prosthetic designs is challenging the clinical and technical expertise of the physician and prosthetist. When generating a prosthetic prescription, physicians and prosthetists should consider the needs and preferences of the athlete with limb deficiency, as well as the functional demands of the chosen sporting activity. The intent of this article is to provide information regarding the current advancements in the adaptive sports technology and biomechanics in the field of prosthetics, and to assist clinicians and their patients in facilitating participation in sporting activities. Copyright © 2014 American Academy of Physical Medicine and Rehabilitation. Published by Elsevier Inc. All rights reserved.

  13. The Prosthetic Experience Between Body and Technology

    DEFF Research Database (Denmark)

    Søndergaard, Morten

    2018-01-01

    In this paper, I argue that a prosthetic aesthetic instigated by experimental art practices operate with and within a ‘second nature’ – in-between science and art. Drawing on theories from Dewey and Edelman and examples from Da Vinci, Brancusi, Man Ray, Dali and Stelarc, I am calling for an exper...... in art, which is performing a complex re-design of (the idea and representation of) technology and the body.......In this paper, I argue that a prosthetic aesthetic instigated by experimental art practices operate with and within a ‘second nature’ – in-between science and art. Drawing on theories from Dewey and Edelman and examples from Da Vinci, Brancusi, Man Ray, Dali and Stelarc, I am calling...... for an experience-based analysis of experimental practices operating between body and technology. These practices, which, rather than falling into the category of science fiction or horror cinema as some recent critique from post-human studies would have it, are pointing towards a genealogy of prosthetic experience...

  14. Stabilization of cytochrome b5 by a conserved tyrosine in the secondary sphere of heme active site: A spectroscopic and computational study

    Science.gov (United States)

    Hu, Shan; He, Bo; Wang, Xiao-Juan; Gao, Shu-Qin; Wen, Ge-Bo; Lin, Ying-Wu

    2017-03-01

    Heme proteins perform a large array of biological functions, with the heme group bound non-covalently or covalently. To probe the stabilization role of conserved tyrosine residue in the secondary sphere of heme site in heme proteins, we herein used cytochrome b5 (Cyt b5) as a model protein, and mutated Tyr30 to Phe or His by removal of Tyr30 associated H-bond network and hydrophobic interaction. We performed thermal-induced unfolding studies for the two mutants, Y30F Cyt b5 and Y30H Cyt b5, as monitored by both UV-Vis and CD spectroscopy, as well as heme transfer studies from these proteins to apo-myoglobin, with wild-type Cyt b5 under the same conditions for comparison. The reduced stability of both mutants indicates that both the H-bonding and hydrophobic interactions associated with Tyr30 contribute to the protein stability. Moreover, we performed molecular modeling studies, which revealed that the hydrophobic interaction in the local region of Y30F Cyt b5 was well-remained, whereas Y30H Cyt b5 formed an H-bond network. These observations suggest that the conserved Tyr30 in Cyt b5 is not replaceable due to the presence of both the H-bond network and hydrophobic interaction in the secondary sphere of the heme active site. As demonstrated here for Cyt b5, it may be of practical importance for design of artificial heme proteins by engineering a Tyr in the secondary sphere with improved properties and functions.

  15. "Look and feel your best": representations of artificial limb users in prosthetic company advertisements.

    Science.gov (United States)

    Murray, Craig D; Forshaw, Mark J

    2014-01-01

    Artificial limbs (prosthetics) are considered important for keeping the person physically active and avoiding an array of negative health outcomes associated with non-use. Increasingly, the potential users of these limbs are the focus of commercial prosthetic company advertisements. It has been argued that it is important to examine such media representations, not least because people's beliefs regarding health and illness are often forged from the discourses and constructions available to them in such material, but because these representations mediate individual lived experience. This article provides a thematic analysis, drawing upon discourse analysis and semiotics, of textual-pictorial representations of artificial limb users in the advertisements of prosthetic companies. The data set was comprised of advertisements that appeared over a 2-year period in inMotion, an international magazine produced and distributed by a major amputee advocacy group. The findings indicate that dominant societal constructions of work, gender and family are drawn on in depicting artificial limb users. These offer generally positive representations that draw on socially pervasive stereotypes. The findings are discussed in relation to literature concerning the experience and meaning of prosthesis use, and the implications for health professionals working with this group are set out. Implications for Rehabilitation People who lose a limb are increasingly being exposed to advertisements from prosthetic companies. Such advertisements have the potential to foster unrealistic expectations regarding rehabilitation following amputation. Healthcare professionals need to be mindful of how these advertisements mediate lived experience and impact on rehabilitation when planning personal care plans.

  16. Effect of a combined anti-thrombotic therapy of thrombosis on prosthetic heart valves.

    Science.gov (United States)

    Wei, Wei; Dong, Taiming; Zheng, Zhichao; Huang, Shuping

    2015-03-01

    To evaluate the curative effects and risks of a medical therapy with combined anti-thrombotic agents for thrombosis on prosthetic heart valves. Twenty-two patients who suffered from thrombosis on prosthetic valves with stable hemodynamics were divided into the inpatient group and the outpatient group. Thrombosis on the valves were demonstrated by transesophageal echocardiographies (TEE). A combined anti-thrombotic therapy with clopidogrel and warfarin were prescribed for all the patients during the whole treatment. Low molecular weight heparin (LMWH) was given twice daily during the first 5 days for the inpatients. The patients accepted regular follow-ups for observation of the functions of prosthetic valves, changes of thrombi, coagulation status and general clinical status. There were 5 men and 17 women. Thirteen patients suffered from thrombosis on the mechanical mitral valves (MVs), five on the mechanical tricuspid valves (TVs), one on the mechanical aortic valve and tricuspid bio-prosthetic valve, one on the mechanical aortic valve, one on the mitral bio-prosthetic valve, and one on the tricuspid bio-prosthetic valve. After an average of 36.4±23.1 days' observation, 16 (73%) patients' valvular function recovered normal without TTE detectable thrombi, 6 (27%) patients' valvular function remained abnormal including three patients without TTE detectable thrombi during follow-ups. No significant differences of thrombi changes and period of thrombi disappearance were observed between the inpatient group and the outpatient group. For patients with mitral thrombosis, sizes of the left atriums (LAs) decreased an average of 4.1 mm after treatment (95% CI, 1.2-6.9 mm). No significant changes of other chambers and left ventricular ejection fractions (LVEF) were observed. For patients with tricuspid thrombosis, LVEF improved an average of 10.5% after treatment (95% CI, 0.1-17.9%). No significant changes of chambers were observed. None experienced major bleedings except

  17. Heme oxygenase activity correlates with serum indices of iron homeostasis in healthy nonsmokers

    Science.gov (United States)

    Heme oxygenase (HO) catalyzes the breakdown of heme to carbon monoxide, iron, and biliverdin. While the use of genetically altered animal models in investigation has established distinct associations between HO activity and systemic iron availability, studies have not yet confirm...

  18. Novel Insights into the Vasoprotective Role of Heme Oxygenase-1

    Directory of Open Access Journals (Sweden)

    Emanuela Marcantoni

    2012-01-01

    Full Text Available Cardiovascular risk factors contribute to enhanced oxidative stress which leads to endothelial dysfunction. These events trigger platelet activation and their interaction with leukocytes and endothelial cells, thus contributing to the induction of chronic inflammatory processes at the vascular wall and to the development of atherosclerotic lesions and atherothrombosis. In this scenario, endogenous antioxidant pathways are induced to restrain the development of vascular disease. In the present paper, we will discuss the role of heme oxygenase (HO-1 which is an enzyme of the heme catabolism and cleaves heme to form biliverdin and carbon monoxide (CO. Biliverdin is reduced enzymatically to the potent antioxidant bilirubin. Recent evidence supports the involvement of HO-1 in the antioxidant and antiinflammatory effect of cyclooxygenase(COX-2-dependent prostacyclin in the vasculature. Moreover, the role of HO-1 in estrogen vasoprotection is emerging. Finally, possible strategies to develop novel therapeutics against cardiovascular disease by targeting the induction of HO-1 will be discussed.

  19. Human heme oxygenase oxidation of 5- and 15-phenylhemes.

    Science.gov (United States)

    Wang, Jinling; Niemevz, Fernando; Lad, Latesh; Huang, Liusheng; Alvarez, Diego E; Buldain, Graciela; Poulos, Thomas L; de Montellano, Paul R Ortiz

    2004-10-08

    Human heme oxygenase-1 (hHO-1) catalyzes the O2-dependent oxidation of heme to biliverdin, CO, and free iron. Previous work indicated that electrophilic addition of the terminal oxygen of the ferric hydroperoxo complex to the alpha-meso-carbon gives 5-hydroxyheme. Earlier efforts to block this reaction with a 5-methyl substituent failed, as the reaction still gave biliverdin IXalpha. Surprisingly, a 15-methyl substituent caused exclusive cleavage at the gamma-meso-rather than at the normal, unsubstituted alpha-meso-carbon. No CO was formed in these reactions, but the fragment cleaved from the porphyrin eluded identification. We report here that hHO-1 cleaves 5-phenylheme to biliverdin IXalpha and oxidizes 15-phenylheme at the alpha-meso position to give 10-phenylbiliverdin IXalpha. The fragment extruded in the oxidation of 5-phenylheme is benzoic acid, one oxygen of which comes from O2 and the other from water. The 2.29- and 2.11-A crystal structures of the hHO-1 complexes with 1- and 15-phenylheme, respectively, show clear electron density for both the 5- and 15-phenyl rings in both molecules of the asymmetric unit. The overall structure of 15-phenylheme-hHO-1 is similar to that of heme-hHO-1 except for small changes in distal residues 141-150 and in the proximal Lys18 and Lys22. In the 5-phenylheme-hHO-1 structure, the phenyl-substituted heme occupies the same position as heme in the heme-HO-1 complex but the 5-phenyl substituent disrupts the rigid hydrophobic wall of residues Met34, Phe214, and residues 26-42 near the alpha-meso carbon. The results provide independent support for an electrophilic oxidation mechanism and support a role for stereochemical control of the reaction regiospecificity.

  20. Control of Prosthetic Hands via the Peripheral Nervous System

    National Research Council Canada - National Science Library

    Ciancio, Anna Lisa; Cordella, Francesca; Barone, Roberto; Romeo, Rocco Antonio; Bellingegni, Alberto Dellacasa; Sacchetti, Rinaldo; Davalli, Angelo; Di Pino, Giovanni; Ranieri, Federico; Di Lazzaro, Vincenzo; Guglielmelli, Eugenio; Zollo, Loredana

    2016-01-01

    ...), and their experimental validation on amputees. The study opens with an in-depth analysis of control solutions and sensorization features of research and commercially available prosthetic hands...

  1. Design of a prosthetic hand with remote actuation.

    Science.gov (United States)

    Scott, Kurt; Perez-Gracia, Alba

    2012-01-01

    One of the main issues of prosthetic hands is to be able to fulfill all the specifications about speed, torque, weight and inertia while placing all the components within the prosthetic hand. This is especially true when full dexterity is required in the prosthesis. In this paper, a new design for a prosthetic hand is presented, which uses remote actuation in order to satisfy most of those requirements. The actuators are to be located in the back of the subject and the transmission is implemented via cables. Other characteristics of this new prosthetic hand include torque limitation and the possibility of switching between underactuated and fully actuated functions.

  2. Factors Associated with Prosthetic Looseness in Lower Limb Amputees.

    Science.gov (United States)

    Phonghanyudh, Thong; Sutpasanon, Taweesak; Hathaiareerug, Chanasak; Devakula, M L Buddhibongsa; Kumnerddee, Wipoo

    2015-12-01

    To determine the factors associated with prosthetic looseness in lower limb amputees in Sisaket province. The present was a cross-sectional descriptive study. Subjects were lower limb amputees who previously obtained prostheses and required prosthetic replacements at the mobile prosthetic laboratory unit under the Prostheses Foundation of H.R.H. the Princess Mother at Khun Han Hospital, Sisaket province, in February 2013. Data including participant characteristics, prosthetic looseness data, and various variables were collected by direct semi-structured interview. Energy expenditures in physical activities were measured using the Thai version of the short format international physical activity questionnaire. Data between participants with and without prosthetic looseness were compared to determine prosthetic loosening associated factors. Among 101 participants enrolled, 33 (32.7%) had prosthetic looseness with average onset of 1.76 ± 1.67 years. Diabetes mellitus was the only significant factor associated with prosthetic looseness from both univariate and multivariate analyses (HR = 7.05, p = 0.002 and HR = 5.93, p = 0.007 respectively). Among the lower limb amputees in Sisaket province, diabetes mellitus was the only factor associated with prosthetic looseness. Therefore, diabetic screening should be supplemented in lower limb amputee assessment protocol. In addition, we recommend that amputees with diabetes mellitus should receive prosthesis check out at approximately

  3. The role of osteoblasts in peri-prosthetic osteolysis.

    LENUS (Irish Health Repository)

    O'Neill, S C

    2013-08-01

    Peri-prosthetic osteolysis and subsequent aseptic loosening is the most common reason for revising total hip replacements. Wear particles originating from the prosthetic components interact with multiple cell types in the peri-prosthetic region resulting in an inflammatory process that ultimately leads to peri-prosthetic bone loss. These cells include macrophages, osteoclasts, osteoblasts and fibroblasts. The majority of research in peri-prosthetic osteolysis has concentrated on the role played by osteoclasts and macrophages. The purpose of this review is to assess the role of the osteoblast in peri-prosthetic osteolysis. In peri-prosthetic osteolysis, wear particles may affect osteoblasts and contribute to the osteolytic process by two mechanisms. First, particles and metallic ions have been shown to inhibit the osteoblast in terms of its ability to secrete mineralised bone matrix, by reducing calcium deposition, alkaline phosphatase activity and its ability to proliferate. Secondly, particles and metallic ions have been shown to stimulate osteoblasts to produce pro inflammatory mediators in vitro. In vivo, these mediators have the potential to attract pro-inflammatory cells to the peri-prosthetic area and stimulate osteoclasts to absorb bone. Further research is needed to fully define the role of the osteoblast in peri-prosthetic osteolysis and to explore its potential role as a therapeutic target in this condition.

  4. Redox properties of lysine- and methionine-coordinated hemes ensure downhill electron transfer in NrfH2A4 nitrite reductase.

    Science.gov (United States)

    Todorovic, Smilja; Rodrigues, Maria Luísa; Matos, Daniela; Pereira, Inês A C

    2012-05-17

    The multiheme NrfHA nitrite reductase is a menaquinol:nitrite oxidoreductase that catalyzes the 6-electron reduction of nitrite to ammonia in a reaction that involves eight protons. X-ray crystallography of the enzyme from Desulfovibrio vulgaris revealed that the biological unit, NrfH2A4, houses 28 c-type heme groups, 22 of them with low spin and 6 with pentacoordinated high spin configuration. The high spin hemes, which are the electron entry and exit points of the complex, carry a highly unusual coordination for c-type hemes, lysine and methionine as proximal ligands in NrfA and NrfH, respectively. Employing redox titrations followed by X-band EPR spectroscopy and surface-enhanced resonance Raman spectroelectrochemistry, we provide the first experimental evidence for the midpoint redox potential of the NrfH menaquinol-interacting methionine-coordinated heme (-270 ± 10 mV, z = 0.96), identified by the use of the inhibitor HQNO, a structural analogue of the physiological electron donor. The redox potential of the catalytic lysine-coordinated high spin heme of NrfA is -50 ± 10 mV, z = 0.9. These values determined for the integral NrfH2A4 complex indicate that a driving force for a downhill electron transfer is ensured in this complex.

  5. Undergraduate prosthetics and orthotics programme objectives:a baseline for international comparison and curricular development.

    Science.gov (United States)

    Aminian, Gholamreza; O'Toole, John Mitchell

    2011-12-01

    Prosthetics and orthotics is a relatively recent addition to the suite of undergraduate professional preparation programmes. There has been limited publication regarding international patterns of curriculum development, particularly concerning how objectives differ across global regions. This paper compares current prosthetics and orthotics curricula from a range of regions and identifies both common and distinctive objectives. Mixed method: document analysis followed by modified Delphi process. Documents were analysed qualitatively to compare various curricula and emergent features were evaluated by a group of expert prosthetics and orthotics instructors. There was substantial agreement that programmes should improve student knowledge and understanding. They should establish and extend student fabrication, communication skills and professional co-operation. However, there appeared to be regional differences in the priority given to critical thinking and clinical reasoning; integration of theory and practice and particular approaches to teaching prosthetics and orthotics. This study revealed substantial consensus regarding the importance of clear programme objectives dealing with student abilities, professional skills and contemporary understanding. However, this study also revealed regional differences that may well reward further investigation.

  6. Graduates' perceptions of prosthetic and orthotic education and clinical practice in Tanzania and Malawi.

    Science.gov (United States)

    Magnusson, Lina; Shangali, Harold G; Ahlström, Gerd

    2016-01-01

    Maintaining and improving the quality of prosthetics and orthotics education at the Tanzania Training Centre for Orthopaedic Technologists is essential for the provision of appropriate prosthetics and orthotics services in African countries. To describe how Tanzanian and Malawian graduates' of the Diploma in Orthopaedic Technology perceive their education and how it could be improved or supplemented to facilitate clinical practice of graduates. Nineteen graduates from the diploma course in orthopaedic technology were interviewed and phenomenographic analysis was applied to the data. Seven descriptive categories emerged, namely varied awareness of the profession before starting education, well-equipped teaching facilities, aspects lacking in the learning context, need for changes in the curriculum, enabling people to walk is motivating, obstacles in working conditions and the need for continuous professional development. All participants perceived possible improvements to the content and learning environment. Prosthetic and orthotic education can be better provided by modifying the content of the diploma programme by dedicating more time to the clinical management of different patient groups and applied biomechanics as well as reducing the programme content focusing on technical aspects of prosthetic and orthotic practice. Graduates were not prepared for the rural working conditions and the graduates desired continued training.

  7. Reconsidering evidence-based practice in prosthetic rehabilitation : a shared enterprise

    NARCIS (Netherlands)

    van Twillert, S.; Geertzen, J.; Hemminga, T.; Postema, K.; Lettinga, A.

    Background: A divide is experienced between producers and users of evidence in prosthetic rehabilitation. Objective: To discuss the complexity inherent in establishing evidence-based practice in a prosthetic rehabilitation team illustrated by the case of prosthetic prescription for elderly

  8. Prosthetic management of a patient with Treacher Collins syndrome

    Directory of Open Access Journals (Sweden)

    Madhan R

    2006-01-01

    Full Text Available Treacher Collins syndrome encompasses a group of closely related defects of the head and neck. It is a rare syndrome characterized by bilaterally symmetrical abnormalities derived from the first and second brachial arches and the nasal placode. It is an autosomal dominant disorder and its occurence ranges from 1 in 25,000 to 1 in 50,000 live births. The facial appearance of these patients can be improved by either surgical or prosthetic rehabilitation. In this case report we are presenting the features of a 13-year-old boy with Treacher Collins syndrome. A multidisplinary approach was followed in managing the situation. The various treatment options and the steps involved in making an auricular prosthesis are also discussed.

  9. Malaria parasite-synthesized heme is essential in the mosquito and liver stages and complements host heme in the blood stages of infection.

    Directory of Open Access Journals (Sweden)

    Viswanathan Arun Nagaraj

    Full Text Available Heme metabolism is central to malaria parasite biology. The parasite acquires heme from host hemoglobin in the intraerythrocytic stages and stores it as hemozoin to prevent free heme toxicity. The parasite can also synthesize heme de novo, and all the enzymes in the pathway are characterized. To study the role of the dual heme sources in malaria parasite growth and development, we knocked out the first enzyme, δ-aminolevulinate synthase (ALAS, and the last enzyme, ferrochelatase (FC, in the heme-biosynthetic pathway of Plasmodium berghei (Pb. The wild-type and knockout (KO parasites had similar intraerythrocytic growth patterns in mice. We carried out in vitro radiolabeling of heme in Pb-infected mouse reticulocytes and Plasmodium falciparum-infected human RBCs using [4-(14C] aminolevulinic acid (ALA. We found that the parasites incorporated both host hemoglobin-heme and parasite-synthesized heme into hemozoin and mitochondrial cytochromes. The similar fates of the two heme sources suggest that they may serve as backup mechanisms to provide heme in the intraerythrocytic stages. Nevertheless, the de novo pathway is absolutely essential for parasite development in the mosquito and liver stages. PbKO parasites formed drastically reduced oocysts and did not form sporozoites in the salivary glands. Oocyst production in PbALASKO parasites recovered when mosquitoes received an ALA supplement. PbALASKO sporozoites could infect mice only when the mice received an ALA supplement. Our results indicate the potential for new therapeutic interventions targeting the heme-biosynthetic pathway in the parasite during the mosquito and liver stages.

  10. Tactile sensing means for prosthetic limbs

    Science.gov (United States)

    Scott, W. L. (Inventor)

    1973-01-01

    An improved prosthetic device characterized by a frame and a socket for mounting on the stump of a truncated human appendage is described. Flexible digits extend from the distal end and transducers located within the digits act as sensing devices for detecting tactile stimuli. The transducers are connected through a power circuit with a slave unit supported by a strap and fixed to the stump. The tactile stimuli detected at the sensing devices are reproduced and applied to the skin of the appendage in order to stimulate the sensory organs located therein.

  11. Smart Prosthetic Hand Technology - Phase 2

    Science.gov (United States)

    2011-05-01

    Belgrade/USC Hand (Bekey, Tomovic and Zeljkovic 1990) has small conductive plastic potentiometers compact (35x10x3mm) with good resolution (320ohms...upper limb loss and their reported research priorities. Journal of Prosthetic and Orthotics 8 (1), 2–11. Bekey, G. A., Tomovic , R., Zeljkovic, I...G.R. Tomovic , and I. Zeljkovic. "Control Architecture for the Belgrade/USC Hand." In Dextrous Robot Hands, by S.T. Venkataram and T. Iberall, 136-149

  12. Injectible bodily prosthetics employing methacrylic copolymer gels

    Energy Technology Data Exchange (ETDEWEB)

    Mallapragada, Surya K.; Anderson, Brian C.

    2007-02-27

    The present invention provides novel block copolymers as structural supplements for injectible bodily prosthetics employed in medical or cosmetic procedures. The invention also includes the use of such block copolymers as nucleus pulposus replacement materials for the treatment of degenerative disc disorders and spinal injuries. The copolymers are constructed by polymerization of a tertiary amine methacrylate with either a (poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) polymer, such as the commercially available Pluronic.RTM. polymers, or a poly(ethylene glycol) methyl ether polymer.

  13. Heme utilization by pathogenic bacteria: not all pathways lead to biliverdin.

    Science.gov (United States)

    Wilks, Angela; Ikeda-Saito, Masao

    2014-08-19

    The eukaryotic heme oxygenases (HOs) (E.C. 1.14.99.3) convert heme to biliverdin, iron, and carbon monoxide (CO) in three successive oxygenation steps. Pathogenic bacteria require iron for survival and infection. Extracellular heme uptake from the host plays a critical role in iron acquisition and virulence. In the past decade, several HOs required for the release of iron from extracellular heme have been identified in pathogenic bacteria, including Corynebacterium diphtheriae, Neisseriae meningitides, and Pseudomonas aeruginosa. The bacterial enzymes were shown to be structurally and mechanistically similar to those of the canonical eukaryotic HO enzymes. However, the recent discovery of the structurally and mechanistically distinct noncanonical heme oxygenases of Staphylococcus aureus and Mycobacterium tuberculosis has expanded the reaction manifold of heme degradation. The distinct ferredoxin-like structural fold and extreme heme ruffling are proposed to give rise to the alternate heme degradation products in the S. aureus and M. tuberculosis enzymes. In addition, several "heme-degrading factors" with no structural homology to either class of HOs have recently been reported. The identification of these "heme-degrading proteins" has largely been determined on the basis of in vitro heme degradation assays. Many of these proteins were reported to produce biliverdin, although no extensive characterization of the products was performed. Prior to the characterization of the canonical HO enzymes, the nonenzymatic degradation of heme and heme proteins in the presence of a reductant such as ascorbate or hydrazine, a reaction termed "coupled oxidation", served as a model for biological heme degradation. However, it was recognized that there were important mechanistic differences between the so-called coupled oxidation of heme proteins and enzymatic heme oxygenation. In the coupled oxidation reaction, the final product, verdoheme, can readily be converted to biliverdin

  14. Heme and HO-1 inhibition of HCV, HBV, and HIV

    Directory of Open Access Journals (Sweden)

    Warren N Schmidt

    2012-10-01

    Full Text Available Hepatitis C virus, human immunodeficiency virus, and hepatitis B virus are chronic viral infections that cause considerable morbidity and mortality throughout the world. In the decades following the identification and sequencing of these viruses, in vitro experiments demonstrated that heme oxygenase-1, its oxidative products, and related compounds of the heme oxygenase system are virucidal for all three viruses. The purpose of this review is to critically evaluate and summarize the seminal studies that described and characterized this remarkable behavior. It will also discuss more recent work that discovered the antiviral mechanisms and target sites of these unique antiviral agents. In spite of the fact that these viruses are diverse pathogens with quite profound differences in structure and life cycle, it is significant that heme and related compounds show striking similarity for viral target sites across all three species. Collectively, these findings strongly indicate that we should move forward and develop heme and related tetrapyrroles into versatile antiviral agents that could be used therapeutically in patients with single or multiple viral infections.

  15. Heme: From quantum spin crossover to oxygen manager of life

    DEFF Research Database (Denmark)

    Kepp, Kasper Planeta

    2016-01-01

    discusses the spectroscopic and computational data that have helped to elucidate the nature of this remarkable molecular system, how it works, and how it is tuned by a range of molecular strategies. This tuning enables heme to carry out the two essential functions required for oxygen management of life, i...

  16. Cysteine-independent activation/inhibition of heme oxygenase-2.

    Science.gov (United States)

    Vukomanovic, Dragic; Rahman, Mona N; Maines, Mahin D; Ozolinš, Terence Rs; Szarek, Walter A; Jia, Zongchao; Nakatsu, Kanji

    2016-03-01

    Reactive thiols of cysteine (cys) residues in proteins play a key role in transforming chemical reactivity into a biological response. The heme oxygenase-2 (HO-2) isozyme contains two cys residues that have been implicated in binding of heme and also the regulation of its activity. In this paper, we address the question of a role for cys residues for the HO-2 inhibitors or activators designed in our laboratory. We tested the activity of full length recombinant human heme oxygenase-2 (FL-hHO-2) and its analog in which cys265 and cys282 were both replaced by alanine to determine the effect on activation by menadione (MD) and inhibition by QC-2350. Similar inhibition by QC-2350 and almost identical activation by MD was observed for both recombinant FL-hHO-2s. Our findings are interpreted to mean that thiols of FL-hHO-2s are not involved in HO-2 activation or inhibition by the compounds that have been designed and identified by us. Activation or inhibition of HO-2 by our compounds should be attributed to a mechanism other than altering binding affinity of HO-2 for heme through cys265 and cys282.

  17. Identification of two genes potentially associated in iron-heme ...

    Indian Academy of Sciences (India)

    2013-03-15

    Mar 15, 2013 ... Classic characteristics are poor predictors of the risk of thromboembolism. Thus, better markers for the carotid atheroma plaque formation and symptom causing are needed. Our objective was to study by microarray analysis gene expression of genes involved in homeostasis of iron and heme in carotid ...

  18. Identification of two genes potentially associated in iron-heme ...

    Indian Academy of Sciences (India)

    Classic characteristics are poor predictors of the risk of thromboembolism. Thus, better markers for the carotid atheroma plaque formation and symptom causing are needed. Our objective was to study by microarray analysis gene expression of genes involved in homeostasis of iron and heme in carotid atheroma plaque ...

  19. Cysteine-independent activation/inhibition of heme oxygenase-2

    Directory of Open Access Journals (Sweden)

    Dragic Vukomanovic

    2016-01-01

    Full Text Available Reactive thiols of cysteine (cys residues in proteins play a key role in transforming chemical reactivity into a biological response. The heme oxygenase-2 (HO-2 isozyme contains two cys residues that have been implicated in binding of heme and also the regulation of its activity. In this paper, we address the question of a role for cys residues for the HO-2 inhibitors or activators designed in our laboratory. We tested the activity of full length recombinant human heme oxygenase-2 (FL-hHO-2 and its analog in which cys265 and cys282 were both replaced by alanine to determine the effect on activation by menadione (MD and inhibition by QC-2350. Similar inhibition by QC-2350 and almost identical activation by MD was observed for both recombinant FL-hHO-2s. Our findings are interpreted to mean that thiols of FL-hHO-2s are not involved in HO-2 activation or inhibition by the compounds that have been designed and identified by us. Activation or inhibition of HO-2 by our compounds should be attributed to a mechanism other than altering binding affinity of HO-2 for heme through cys265 and cys282.

  20. Age-related accumulation of non-heme ferric and ferrous iron in mouse ovarian stroma visualized by sensitive non-heme iron histochemistry.

    Science.gov (United States)

    Asano, Yoshiya

    2012-03-01

    Sensitive non-heme iron histochemistry--namely, the perfusion-Perls method and perfusion-Turnbull method--was applied to study the distribution and age-related accumulation of non-heme ferric iron and ferrous iron in mouse ovary. Light and electron microscopic studies revealed that non-heme ferric iron is distributed predominantly in stromal tissue, especially in macrophages. By contrast, the distribution of non-heme ferrous iron was restricted to a few ovoid macrophages. Aged ovaries exhibited remarkable non-heme iron accumulation in all stromal cells. In particular, non-heme ferrous iron level was increased in stromal tissue, suggestive of increased levels of redox-active iron, which can promote oxidative stress. Moreover, intense localization of both non-heme ferric and ferrous iron was observed in aggregated large stromal cells that were then characterized as ceroid-laden enlarged macrophages with frothy cytoplasm. Intraperitoneal iron overload in adult mice resulted in non-heme iron deposition in the entire stroma and generation of enlarged macrophages, suggesting that excessive iron accumulation induced macrophage morphological changes. The data indicated that non-heme iron accumulation in ovarian stromal tissue may be related to aging of the ovary due to increasing oxidative stress.

  1. Contribution of prosthetic treatment considerations for dental extractions of permanent teeth

    Directory of Open Access Journals (Sweden)

    Miguel Ángel Fernández-Barrera

    2016-07-01

    Full Text Available Background. Tooth loss is an easily identifiable outcome that summarizes a complex suite of factors in an individual’s history of dental disease and its treatment by dental services over a lifetime. Assessment of overall tooth loss data is essential for epidemiologically evaluating the adequacy of dental care provided at a systems level, as well as for placing in context tooth loss for non-disease causes. For example, when derived from prosthetic treatment planning, the latter may unfortunately lead to some teeth being extracted (pulled for the sake of better comprehensive clinical results. The objective of the present manuscript was to identify the contribution to overall tooth loss, by extraction of permanent teeth because of prosthetic treatment reasons. Material and Methods. A cross-sectional study included sex, age, total number of extractions performed by subject, sextant (anterior vs. posterior, group of teeth (incisors, canines, premolars and molars, upper or lower arch, and the main reason underlying extraction (extraction for any reason vs. prosthetic treatment, in patients 18 years of age and older seeking care at a dental school clinic in Mexico. A multivariate logistic regression model was generated. Results. A total of 749 teeth were extracted in 331 patients; 161 teeth (21.5% of total were extracted for explicit prosthetic treatment indications. As age increased, the likelihood of having an extraction for prosthetic reasons increased 3% (OR = 1.03, p < 0.001. Women (OR = 1.57, p < 0.05 were more likely to be in this situation, and molars (OR = 2.70, p < 0.001 were most at risk. As the total number of extractions increased, the risk of having an extraction for prosthetic reasons decreased (OR = 0.94, p < 0.05. Conclusions. A significant amount (21.5% of the extractions of permanent teeth were performed for prosthetic reasons in this dental school clinical environment; age, sex, type of tooth, and the total number of extractions

  2. Identification and characterization of a heme periplasmic-binding protein in Haemophilus ducreyi.

    Science.gov (United States)

    St Denis, Melissa; Sonier, Brigitte; Robinson, Renée; Scott, Fraser W; Cameron, D William; Lee, B Craig

    2011-08-01

    Haemophilus ducreyi, a gram-negative and heme-dependent bacterium, is the causative agent of chancroid, a genital ulcer sexually transmitted infection. Heme acquisition in H. ducreyi proceeds via a receptor mediated process in which the initial event involves binding of hemoglobin and heme to their cognate outer membrane proteins, HgbA and TdhA, respectively. Following this specific interaction, the fate of the periplasmic deposited heme is unclear. Using protein expression profiling of the H. ducreyi periplasmic proteome, a periplasmic-binding protein, termed hHbp, was identified whose expression was enhanced under heme-limited conditions. The gene encoding this protein was situated in a locus displaying genetic characteristics of an ABC transporter. The purified protein bound heme in a dose-dependent and saturable manner and this binding was specifically competitively inhibited by heme. The hhbp gene functionally complemented an Escherichia coli heme uptake mutant. Expression of the heme periplasmic-binding protein was detected in a limited survey of H. ducreyi and H. influenzae clinical strains. These results indicate that the passage of heme into the cytoplasm of H. ducreyi involves a heme dedicated ABC transporter.

  3. 38 CFR 17.150 - Prosthetic and similar appliances.

    Science.gov (United States)

    2010-07-01

    ... appliances. 17.150 Section 17.150 Pensions, Bonuses, and Veterans' Relief DEPARTMENT OF VETERANS AFFAIRS MEDICAL Prosthetic, Sensory, and Rehabilitative Aids § 17.150 Prosthetic and similar appliances. Artificial limbs, braces, orthopedic shoes, hearing aids, wheelchairs, medical accessories, similar...

  4. Prosthetic Frequently Asked Questions for the New Amputee

    Science.gov (United States)

    ... it with your favorite color or pattern. The prosthesis is an extension of you and your style – wear it proudly! Technology continues to change the prosthetic market. With advances in the microprocessor knee and foot, and advanced hands and sockets, prosthetics ...

  5. Prosthetic Rehabilitation in Children: An Alternative Clinical Technique

    Directory of Open Access Journals (Sweden)

    Nádia Carolina Teixeira Marques

    2013-01-01

    Full Text Available Complete and partial removable dentures have been used successfully in numerous patients with oligodontia and/or anodontia. However, there is little information in the literature regarding the principles and guidelines to prosthetic rehabilitation for growing children. This case report describes the management of a young child with oligodontia as well as the treatment planning and the prosthetic rehabilitation technique.

  6. Principles of obstacle avoidance with a transfemoral prosthetic limb

    NARCIS (Netherlands)

    van Keeken, Helco G.; Vrieling, Aline H.; Hof, At L.; Postema, Klaas; Otten, Bert

    2012-01-01

    In this study, conditions that enable a prosthetic knee flexion strategy in transfemoral amputee subjects during obstacle avoidance were investigated. This study explored the hip torque principle and the static ground principle as object avoidance strategies. A prosthetic limb simulator device was

  7. Stiffness and hysteresis properties of some prosthetic feet

    NARCIS (Netherlands)

    van Jaarsveld, H.W.L.; Grootenboer, H.J.; de Vries, J.; Koopman, Hubertus F.J.M.

    1990-01-01

    A prosthetic foot is an important element of a prosthesis, although it is not always fully recognized that the properties of the foot, along with the prosthetic knee joint and the socket, are in part responsible for the stability and metabolic energy cost during walking. The stiffness and the

  8. Bioavailability of a heme-iron concentrate product added to chocolate biscuit filling in adolescent girls living in a rural area of Mexico.

    Science.gov (United States)

    González-Rosendo, Guillermina; Polo, Javier; Rodríguez-Jerez, José Juan; Puga-Díaz, Rubén; Reyes-Navarrete, Eduardo G; Quintero-Gutiérrez, Adrián G

    2010-04-01

    A heme-iron concentrate product derived from swine hemoglobin was used to enrich the chocolate-flavored filling of biscuits and the bioavailability of this source of heme-iron was assessed in adolescent girls. The placebo control (PC) group consisted of 35 teenagers with the highest baseline hemoglobin concentrations. The supplemented groups were randomized to receive biscuits fortified with iron sulfate (IS, n = 37) or heme-iron concentrate (HIC, n = 40). Both groups were supplemented with 10.3 mg Fe/d for 7 wk. Blood chemistry and hematology analyses were performed at baseline and at the end of the study. The baseline prevalence of anemia (hemoglobin <12 g/dl) in the entire group was 3.9% and by the end of the study it had fallen to 2.3%. The hemoglobin levels in both supplemented groups increased (P < 0.05) during the study period from 13.6 and 13.5 g/dl for HIC and IS, respectively, at baseline to 14 g/dl at the end of the study. Serum ferritin concentrations decreased by the end of the study in both the PC and IS groups (P < 0.05), but not in the heme group. In conclusion, iron bioavailability from HIC-fortified biscuits was calculated to be 23.7% higher than that observed for IS, as shown by the differences observed in serum ferritin levels during the study. The iron contained in the heme-iron concentrate was well absorbed and tolerated by the adolescents included in the study.

  9. Prosthetic implantation of the human vestibular system.

    Science.gov (United States)

    Golub, Justin S; Ling, Leo; Nie, Kaibao; Nowack, Amy; Shepherd, Sarah J; Bierer, Steven M; Jameyson, Elyse; Kaneko, Chris R S; Phillips, James O; Rubinstein, Jay T

    2014-01-01

    A functional vestibular prosthesis can be implanted in human such that electrical stimulation of each semicircular canal produces canal-specific eye movements while preserving vestibular and auditory function. A number of vestibular disorders could be treated with prosthetic stimulation of the vestibular end organs. We have previously demonstrated in rhesus monkeys that a vestibular neurostimulator, based on the Nucleus Freedom cochlear implant, can produce canal-specific electrically evoked eye movements while preserving auditory and vestibular function. An investigational device exemption has been obtained from the FDA to study the feasibility of treating uncontrolled Ménière's disease with the device. The UW/Nucleus vestibular implant was implanted in the perilymphatic space adjacent to the three semicircular canal ampullae of a human subject with uncontrolled Ménière's disease. Preoperative and postoperative vestibular and auditory function was assessed. Electrically evoked eye movements were measured at 2 time points postoperatively. Implantation of all semicircular canals was technically feasible. Horizontal canal and auditory function were largely, but not totally, lost. Electrode stimulation in 2 of 3 canals resulted in canal-appropriate eye movements. Over time, stimulation thresholds increased. Prosthetic implantation of the semicircular canals in humans is technically feasible. Electrical stimulation resulted in canal-specific eye movements, although thresholds increased over time. Preservation of native auditory and vestibular function, previously observed in animals, was not demonstrated in a single subject with advanced Ménière's disease.

  10. Is the prosthetic homologue necessary for embodiment?

    Directory of Open Access Journals (Sweden)

    Chelsea Dornfeld

    2016-12-01

    Full Text Available Embodiment is the process by which patients with limb loss come to accept their peripheral device as a natural extension of self. However, there is little guidance as to how exacting the prosthesis must be in order for embodiment to take place: is it necessary for the prosthetic hand to look just like the absent hand? Here, we describe a protocol for testing whether an individual would select a hand that looks like their own from among a selection of 5 hands, and whether the hand selection (regardless of homology is consistent across multiple exposures to the same (but reordered set of candidate hands. Pilot results using healthy volunteers reveals that hand selection is only modestly consistent, and that selection of the prosthetic homologue is atypical (61 of 192 total exposures. Our protocol can be executed in minutes, and makes use of readily available equipment and softwares. We present both a face-to-face and a virtual protocol, for maximum flexibility of implementation.

  11. The use of underactuation in prosthetic grasping

    Directory of Open Access Journals (Sweden)

    P. J. Kyberd

    2011-02-01

    Full Text Available Underactuation as a method of driving prosthetic hands has a long history. The pragmatic requirements of such a device to be light enough to be worn and used regularly have meant that any multi degree of freedom prosthetic hand must have fewer actuators than the usable degrees of freedom. Aesthetics ensures that while the hand needs five fingers, five actuators have considerable mass, and only in recent years has it even been possible to construct a practical anthropomorphic hand with five motors. Thus there is an important trade off as to which fingers are driven, and which joints on which fingers are actuated, and how the forces are distributed to create a functional device. This paper outlines some of the historical solutions created for this problem and includes those designs of recent years that are now beginning to be used in the commercial environment.

    This paper was presented at the IFToMM/ASME International Workshop on Underactuated Grasping (UG2010, 19 August 2010, Montréal, Canada.

  12. The Prosthetic Workflow in the Digital Era

    Directory of Open Access Journals (Sweden)

    Lidia Tordiglione

    2016-01-01

    Full Text Available The purpose of this retrospective study was to clinically evaluate the benefits of adopting a full digital workflow for the implementation of fixed prosthetic restorations on natural teeth. To evaluate the effectiveness of these protocols, treatment plans were drawn up for 15 patients requiring rehabilitation of one or more natural teeth. All the dental impressions were taken using a Planmeca PlanScan® (Planmeca OY, Helsinki, Finland intraoral scanner, which provided digital casts on which the restorations were digitally designed using Exocad® (Exocad GmbH, Germany, 2010 software and fabricated by CAM processing on 5-axis milling machines. A total of 28 single crowns were made from monolithic zirconia, 12 vestibular veneers from lithium disilicate, and 4 three-quarter vestibular veneers with palatal extension. While the restorations were applied, the authors could clinically appreciate the excellent match between the digitally produced prosthetic design and the cemented prostheses, which never required any occlusal or proximal adjustment. Out of all the restorations applied, only one exhibited premature failure and was replaced with no other complications or need for further scanning. From the clinical experience gained using a full digital workflow, the authors can confirm that these work processes enable the fabrication of clinically reliable restorations, with all the benefits that digital methods bring to the dentist, the dental laboratory, and the patient.

  13. The Prosthetic Workflow in the Digital Era.

    Science.gov (United States)

    Tordiglione, Lidia; De Franco, Michele; Bosetti, Giovanni

    2016-01-01

    The purpose of this retrospective study was to clinically evaluate the benefits of adopting a full digital workflow for the implementation of fixed prosthetic restorations on natural teeth. To evaluate the effectiveness of these protocols, treatment plans were drawn up for 15 patients requiring rehabilitation of one or more natural teeth. All the dental impressions were taken using a Planmeca PlanScan® (Planmeca OY, Helsinki, Finland) intraoral scanner, which provided digital casts on which the restorations were digitally designed using Exocad® (Exocad GmbH, Germany, 2010) software and fabricated by CAM processing on 5-axis milling machines. A total of 28 single crowns were made from monolithic zirconia, 12 vestibular veneers from lithium disilicate, and 4 three-quarter vestibular veneers with palatal extension. While the restorations were applied, the authors could clinically appreciate the excellent match between the digitally produced prosthetic design and the cemented prostheses, which never required any occlusal or proximal adjustment. Out of all the restorations applied, only one exhibited premature failure and was replaced with no other complications or need for further scanning. From the clinical experience gained using a full digital workflow, the authors can confirm that these work processes enable the fabrication of clinically reliable restorations, with all the benefits that digital methods bring to the dentist, the dental laboratory, and the patient.

  14. Dietary heme mediated PPARα activation does not affect the heme-induced epithelial hyperproliferation and hyperplasia in mouse colon

    NARCIS (Netherlands)

    IJssennagger, Noortje; Wit, de Nicole; Muller, Michael; Meer, van der Roelof

    2012-01-01

    Red meat consumption is associated with an increased colon cancer risk. Heme, present in red meat, injures the colon surface epithelium by luminal cytotoxicity and reactive oxygen species. This surface injury is overcompensated by hyperproliferation and hyperplasia of crypt cells. Transcriptome

  15. Dietary heme-mediated PPARa activation does not affect the heme-induced epithelial hyperproliferation and hyperplasia in mouse colon

    NARCIS (Netherlands)

    IJssenagger, N.; Wit, de N.J.W.; Muller, M.R.; Meer, van der R.

    2012-01-01

    Red meat consumption is associated with an increased colon cancer risk. Heme, present in red meat, injures the colon surface epithelium by luminal cytotoxicity and reactive oxygen species. This surface injury is overcompensated by hyperproliferation and hyperplasia of crypt cells. Transcriptome

  16. Bacterial Nitric Oxide Synthase Is Required for the Staphylococcus aureus Response to Heme Stress.

    Science.gov (United States)

    Surdel, Matthew C; Dutter, Brendan F; Sulikowski, Gary A; Skaar, Eric P

    2016-08-12

    Staphylococcus aureus is a pathogen that causes significant morbidity and mortality worldwide. Within the vertebrate host, S. aureus requires heme as a nutrient iron source and as a cofactor for multiple cellular processes. Although required for pathogenesis, excess heme is toxic. S. aureus employs a two-component system, the heme sensor system (HssRS), to sense and protect against heme toxicity. Upon activation, HssRS induces the expression of the heme-regulated transporter (HrtAB), an efflux pump that alleviates heme toxicity. The ability to sense and respond to heme is critical for the pathogenesis of numerous Gram-positive organisms, yet the mechanism of heme sensing remains unknown. Compound '3981 was identified in a high-throughput screen as an activator of staphylococcal HssRS that triggers HssRS independently of heme accumulation. '3981 is toxic to S. aureus; however, derivatives of '3981 were synthesized that lack toxicity while retaining HssRS activation, enabling the interrogation of the heme stress response without confounding toxic effects of the parent molecule. Using '3981 derivatives as probes of the heme stress response, numerous genes required for '3981-induced activation of HssRS were uncovered. Specifically, multiple genes involved in the production of nitric oxide were identified, including the gene encoding bacterial nitric oxide synthase (bNOS). bNOS protects S. aureus from oxidative stress imposed by heme. Taken together, this work identifies bNOS as crucial for the S. aureus heme stress response, providing evidence that nitric oxide synthesis and heme sensing are intertwined.

  17. A Novel, “Double-Clamp” Binding Mode for Human Heme Oxygenase-1 Inhibition

    Science.gov (United States)

    Rahman, Mona N.; Vlahakis, Jason Z.; Vukomanovic, Dragic; Lee, Wallace; Szarek, Walter A.; Nakatsu, Kanji; Jia, Zongchao

    2012-01-01

    The development of heme oxygenase (HO) inhibitors is critical in dissecting and understanding the HO system and for potential therapeutic applications. We have established a program to design and optimize HO inhibitors using structure-activity relationships in conjunction with X-ray crystallographic analyses. One of our previous complex crystal structures revealed a putative secondary hydrophobic binding pocket which could be exploited for a new design strategy by introducing a functional group that would fit into this potential site. To test this hypothesis and gain further insights into the structural basis of inhibitor binding, we have synthesized and characterized 1-(1H-imidazol-1-yl)-4,4-diphenyl-2-butanone (QC-308). Using a carbon monoxide (CO) formation assay on rat spleen microsomes, the compound was found to be ∼15 times more potent (IC50 = 0.27±0.07 µM) than its monophenyl analogue, which is already a potent compound in its own right (QC-65; IC50 = 4.0±1.8 µM). The crystal structure of hHO-1 with QC-308 revealed that the second phenyl group in the western region of the compound is indeed accommodated by a definitive secondary proximal hydrophobic pocket. Thus, the two phenyl moieties are each stabilized by distinct hydrophobic pockets. This “double-clamp” binding offers additional inhibitor stabilization and provides a new route for improvement of human heme oxygenase inhibitors. PMID:22276118

  18. A novel, "double-clamp" binding mode for human heme oxygenase-1 inhibition.

    Science.gov (United States)

    Rahman, Mona N; Vlahakis, Jason Z; Vukomanovic, Dragic; Lee, Wallace; Szarek, Walter A; Nakatsu, Kanji; Jia, Zongchao

    2012-01-01

    The development of heme oxygenase (HO) inhibitors is critical in dissecting and understanding the HO system and for potential therapeutic applications. We have established a program to design and optimize HO inhibitors using structure-activity relationships in conjunction with X-ray crystallographic analyses. One of our previous complex crystal structures revealed a putative secondary hydrophobic binding pocket which could be exploited for a new design strategy by introducing a functional group that would fit into this potential site. To test this hypothesis and gain further insights into the structural basis of inhibitor binding, we have synthesized and characterized 1-(1H-imidazol-1-yl)-4,4-diphenyl-2-butanone (QC-308). Using a carbon monoxide (CO) formation assay on rat spleen microsomes, the compound was found to be ∼15 times more potent (IC(50) = 0.27±0.07 µM) than its monophenyl analogue, which is already a potent compound in its own right (QC-65; IC(50) = 4.0±1.8 µM). The crystal structure of hHO-1 with QC-308 revealed that the second phenyl group in the western region of the compound is indeed accommodated by a definitive secondary proximal hydrophobic pocket. Thus, the two phenyl moieties are each stabilized by distinct hydrophobic pockets. This "double-clamp" binding offers additional inhibitor stabilization and provides a new route for improvement of human heme oxygenase inhibitors.

  19. A novel, "double-clamp" binding mode for human heme oxygenase-1 inhibition.

    Directory of Open Access Journals (Sweden)

    Mona N Rahman

    Full Text Available The development of heme oxygenase (HO inhibitors is critical in dissecting and understanding the HO system and for potential therapeutic applications. We have established a program to design and optimize HO inhibitors using structure-activity relationships in conjunction with X-ray crystallographic analyses. One of our previous complex crystal structures revealed a putative secondary hydrophobic binding pocket which could be exploited for a new design strategy by introducing a functional group that would fit into this potential site. To test this hypothesis and gain further insights into the structural basis of inhibitor binding, we have synthesized and characterized 1-(1H-imidazol-1-yl-4,4-diphenyl-2-butanone (QC-308. Using a carbon monoxide (CO formation assay on rat spleen microsomes, the compound was found to be ∼15 times more potent (IC(50 = 0.27±0.07 µM than its monophenyl analogue, which is already a potent compound in its own right (QC-65; IC(50 = 4.0±1.8 µM. The crystal structure of hHO-1 with QC-308 revealed that the second phenyl group in the western region of the compound is indeed accommodated by a definitive secondary proximal hydrophobic pocket. Thus, the two phenyl moieties are each stabilized by distinct hydrophobic pockets. This "double-clamp" binding offers additional inhibitor stabilization and provides a new route for improvement of human heme oxygenase inhibitors.

  20. Ibuprofen Impairs Allosterically Peroxynitrite Isomerization by Ferric Human Serum Heme-Albumin*

    Science.gov (United States)

    Ascenzi, Paolo; di Masi, Alessandra; Coletta, Massimo; Ciaccio, Chiara; Fanali, Gabriella; Nicoletti, Francesco P.; Smulevich, Giulietta; Fasano, Mauro

    2009-01-01

    Human serum albumin (HSA) participates in heme scavenging; in turn, heme endows HSA with myoglobin-like reactivity and spectroscopic properties. Here, the allosteric effect of ibuprofen on peroxynitrite isomerization to NO3− catalyzed by ferric human serum heme-albumin (HSA-heme-Fe(III)) is reported. Data were obtained at 22.0 °C. HSA-heme-Fe(III) catalyzes peroxynitrite isomerization in the absence and presence of CO2; the values of the second order catalytic rate constant (kon) are 4.1 × 105 and 4.5 × 105 m−1 s−1, respectively. Moreover, HSA-heme-Fe(III) prevents peroxynitrite-mediated nitration of free added l-tyrosine. The pH dependence of kon (pKa = 6.9) suggests that peroxynitrous acid reacts preferentially with the heme-Fe(III) atom, in the absence and presence of CO2. The HSA-heme-Fe(III)-catalyzed isomerization of peroxynitrite has been ascribed to the reactive pentacoordinated heme-Fe(III) atom. In the absence and presence of CO2, ibuprofen impairs dose-dependently peroxynitrite isomerization by HSA-heme-Fe(III) and facilitates the nitration of free added l-tyrosine; the value of the dissociation equilibrium constant for ibuprofen binding to HSA-heme-Fe(III) (L) ranges between 7.7 × 10−4 and 9.7 × 10−4 m. Under conditions where [ibuprofen] is ≫L, the kinetics of HSA-heme-Fe(III)-catalyzed isomerization of peroxynitrite is superimposable to that obtained in the absence of HSA-heme-Fe(III) or in the presence of non-catalytic HSA-heme-Fe(III)-cyanide complex and HSA. Ibuprofen binding impairs allosterically peroxynitrite isomerization by HSA-heme-Fe(III), inducing the hexacoordination of the heme-Fe(III) atom. These results represent the first evidence for peroxynitrite isomerization by HSA-heme-Fe(III), highlighting the allosteric modulation of HSA-heme-Fe(III) reactivity by heterotropic interaction(s), and outlining the role of drugs in modulating HSA functions. The present results could be relevant for the drug-dependent protective role

  1. Transfection of the Human Heme Oxygenase Gene Into Rabbit Coronary Microvessel Endothelial Cells: Protective Effect Against Heme and Hemoglobin Toxicity

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    Abraham, N. G.; Lavrovsky, Y.; Schwartzman, M. L.; Stoltz, R. A.; Levere, R. D.; Gerritsen, M. E.

    1995-07-01

    Heme oxygenase (HO) is a stress protein and has been suggested to participate in defense mechanisms against agents that may induce oxidative injury such as metals, endotoxin, heme/hemoglobin, and various cytokines. Overexpression of HO in cells might therefore protect against oxidative stress produced by certain of these agents, specifically heme and hemoglobin, by catalyzing their degradation to bilirubin, which itself has antioxidant properties. We report here the successful in vitro transfection of rabbit coronary microvessel endothelial cells with a functioning gene encoding the human HO enzyme. A plasmid containing the cytomegalovirus promoter and the human HO cDNA complexed to cationic liposomes (Lipofectin) was used to transfect rabbit endothelial cells. Cells transfected with human HO exhibited an ≈3.0-fold increase in enzyme activity and expressed a severalfold induction of human HO mRNA as compared with endogenous rabbit HO mRNA. Transfected and nontransfected cells expressed factor VIII antigen and exhibited similar acetylated low-density lipoprotein uptake (two important features that characterize endothelial cells) with >85% of cells staining positive for each marker. Moreover, cells transfected with the human HO gene acquired substantial resistance to toxicity produced by exposure to recombinant hemoglobin and heme as compared with nontransfected cells. The protective effect of HO overexpression against heme/hemoglobin toxicity in endothelial cells shown in these studies provides direct evidence that the inductive response of human HO to such injurious stimuli represents an important tissue adaptive mechanism for moderating the severity of cell damage produced by these blood components.

  2. Heme oxygenase-2 gene deletion attenuates oxidative stress in neurons exposed to extracellular hemin

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    Benvenisti-Zarom Luna

    2004-09-01

    Full Text Available Abstract Background Hemin, the oxidized form of heme, accumulates in intracranial hematomas and is a potent oxidant. Growing evidence suggests that it contributes to delayed injury to surrounding tissue, and that this process is affected by the heme oxygenase enzymes. In a prior study, heme oxygenase-2 gene deletion increased the vulnerability of cultured cortical astrocytes to hemin. The present study tested the effect of HO-2 gene deletion on protein oxidation, reactive oxygen species formation, and cell viability after mixed cortical neuron/astrocyte cultures were incubated with neurotoxic concentrations of hemin. Results Continuous exposure of wild-type cultures to 1–10 μM hemin for 14 h produced concentration-dependent neuronal death, as detected by both LDH release and fluorescence intensity after propidium iodide staining, with an EC50 of 1–2 μM; astrocytes were not injured by these low hemin concentrations. Cell death was consistently reduced by at least 60% in knockout cultures. Exposure to hemin for 4 hours, a time point that preceded cell lysis, increased protein oxidation in wild-type cultures, as detected by staining of immunoblots for protein carbonyl groups. At 10 μM hemin, carbonylation was increased 2.3-fold compared with control sister cultures subjected to medium exchanges only; this effect was reduced by about two-thirds in knockout cultures. Cellular reactive oxygen species, detected by fluorescence intensity after dihydrorhodamine 123 (DHR staining, was markedly increased by hemin in wild-type cultures and was localized to neuronal cell bodies and processes. In contrast, DHR fluorescence intensity in knockout cultures did not differ from that of sham-washed controls. Neuronal death in wild-type cultures was almost completely prevented by the lipid-soluble iron chelator phenanthroline; deferoxamine had a weaker but significant effect. Conclusions These results suggest that HO-2 gene deletion protects neurons in mixed

  3. Metabolite-driven Regulation of Heme Uptake by the Biliverdin IXβ/δ-Selective Heme Oxygenase (HemO) of Pseudomonas aeruginosa.

    Science.gov (United States)

    Mouriño, Susana; Giardina, Bennett J; Reyes-Caballero, Hermes; Wilks, Angela

    2016-09-23

    Pseudomonas aeruginosa acquires extracellular heme via the Phu (Pseudomonas heme uptake) and Has (heme assimilation system) systems. We have previously shown the catalytic actions of heme oxygenase (HemO) along with the cytoplasmic heme transport protein PhuS control heme flux into the cell. To further investigate the role of the PhuS-HemO couple in modulating heme uptake, we have characterized two HemO variants, one that is catalytically inactive (HemO H26A/K34A/K132A or HemOin) and one that has altered regioselectivity (HemO N19K/K34A/F117Y/K132A or HemOα), producing biliverdin IXα (BVIXα). HemOα similar to wild type was able to interact and acquire heme from holo-PhuS. In contrast, the HemOin variant did not interact with holo-PhuS and showed no enzymatic activity. Complementation of a hemO deletion strain with the hemOin or hemOα variants in combination with [(13)C]heme isotopic labeling experiments revealed that the absence of BVIXβ and BVIXδ leads to a decrease in extracellular levels of hemophore HasA. We propose BVIXβ and/or BVIXδ transcriptionally or post-transcriptionally regulates HasA. Thus, coupling the PhuS-dependent flux of heme through HemO to feedback regulation of the cell surface signaling system through HasA allows P. aeruginosa to rapidly respond to fluctuating extracellular heme levels independent of the iron status of the cell. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Structure of the heme/hemoglobin outer membrane receptor ShuA from Shigella dysenteriae: heme binding by an induced fit mechanism.

    Science.gov (United States)

    Cobessi, David; Meksem, Ahmed; Brillet, Karl

    2010-02-01

    Shigella dysentriae and other Gram-negative human pathogens are able to use iron from heme bound to hemoglobin for growing. We solved at 2.6 A resolution the 3D structure of the TonB-dependent heme/hemoglobin outer membrane receptor ShuA from S. dysenteriae. ShuA binds to hemoglobin and transports heme across the outer membrane. The structure consists of a C-terminal domain that folds into a 22-stranded transmembrane beta-barrel, which is filled by the N-terminal plug domain. One distal histidine ligand of heme is located at the apex of the plug, exposed to the solvent. His86 is situated 9.86 A apart from His420, the second histidine involved in the heme binding. His420 is in the extracellular loop L7. The heme coordination by His86 and His420 involves conformational changes. The comparisons with the hemophore receptor HasR of Serratia marcescens bound to HasA-Heme suggest an extracellular induced fit mechanism for the heme binding. The loop L7 contains hydrophobic residues which could interact with the hydrophobic porphyring ring of heme. The energy required for the transport by ShuA is derived from the proton motive force after interactions between the periplasmic N-terminal TonB-box of ShuA and the inner membrane protein, TonB. In ShuA, the TonB-box is buried and cannot interact with TonB. The structural comparisons with HasR suggest its conformational change upon the heme binding for interacting with TonB. The signaling of the heme binding could involve a hydrogen bond network going from His86 to the TonB-box. (c) 2009 Wiley-Liss, Inc.

  5. Effects of kinematic vibrotactile feedback on learning to control a virtual prosthetic arm.

    Science.gov (United States)

    Hasson, Christopher J; Manczurowsky, Julia

    2015-03-24

    After a limb is lost a prosthesis can restore function. For maximum utility, prosthetic limbs should accept movement commands and provide force and motion feedback, which can be conveyed with vibrotactile feedback (VIBF). While prior studies have shown that force-based VIBF benefits control, the merits of motion-based VIBF are unclear. Our goal was to clarify the effectiveness of position- and velocity-based VIBF for prosthetic arm control. Healthy adults with normal limb function practiced a goal-directed task with a virtual myoelectric prosthetic arm. A linear resonant actuator on the wrist provided VIBF. Two groups with nine subjects each received amplitude modulated VIBF in addition to visual feedback while practicing the task. In one group, the VIBF was proportional to the virtual arm's position, and in the other group, velocity. A control group of nine subjects received only visual feedback. Subjects practiced for 240 trials, followed by 180 trials with feedback manipulations for the VIBF groups. Performance was characterized by end-point error, movement time, and a composite skill measure that combined these quantities. A second experiment with a new group of five subjects assessed discrimination capabilities between different position- and velocity-based VIBF profiles. With practice all groups improved their skill in controlling the virtual prosthetic arm. Subjects who received additional position- and velocity-based VIBF learned at the same rate as the control group, who received only visual feedback (learning rate time constant: about 40 trials). When visual feedback was subsequently removed leaving only VIBF, performance was no better than with no feedback at all. When VIBF was removed leaving only visual feedback, about half of the participants performed better, instead of worse. The VIBF discrimination tests showed that subjects could detect virtual arm angular position and velocity differences of about 5 deg and 20 deg/s, respectively. Kinematic

  6. Interaction of nitric oxide with human heme oxygenase-1.

    Science.gov (United States)

    Wang, Jinling; Lu, Shen; Moënne-Loccoz, Pierre; Ortiz de Montellano, Paul R

    2003-01-24

    NO and CO may complement each other as signaling molecules in some physiological situations. We have examined the binding of NO to human heme oxygenase-1 (hHO-1), an enzyme that oxidizes heme to biliverdin, CO, and free iron, to determine whether inhibition of hHO-1 by NO can contribute to the signaling interplay of NO and CO. An Fe(3+)-NO hHO-1-heme complex is formed with NO or the NO donors NOC9 or 2-(N,N-diethylamino)-diazenolate-2-oxide.sodium salt. Resonance Raman spectroscopy shows that ferric hHO-1-heme forms a 6-coordinated, low spin complex with NO. The nu(N-O) vibration of this complex detected by Fourier transform IR is only 4 cm(-1) lower than that of the corresponding metmyoglobin (met-Mb) complex but is broader, suggesting a greater degree of ligand conformational freedom. The Fe(3+)-NO complex of hHO-1 is much more stable than that of met-Mb. Stopped-flow studies indicate that k(on) for formation of the hHO-1-heme Fe(3+)-NO complex is approximately 50-times faster, and k(off) 10 times slower, than for met-Mb, resulting in K(d) = 1.4 microm for NO. NO thus binds 500-fold more tightly to ferric hHO-1-heme than to met-Mb. The hHO-1 mutations E29A, G139A, D140A, S142A, G143A, G143F, and K179A/R183A do not significantly diminish the tight binding of NO, indicating that NO binding is not highly sensitive to mutations of residues that normally stabilize the distal water ligand. As expected from the K(d) value, the enzyme is reversibly inhibited upon exposure to pathologically, and possibly physiologically, relevant concentrations of NO. Inhibition of hHO-1 by NO may contribute to the pleiotropic responses to NO and CO.

  7. A COMPARATIVE STUDY ON THE EFFECTS OF THE POLYMER PROSTHETIC BASE PRESSURE OVER THE PROSTHETIC FIELD

    Directory of Open Access Journals (Sweden)

    A.T. Cigu

    2016-04-01

    Full Text Available Partially mobile polymeric prostheses constitute absolutely necessary therapeutical means in all forms of partial edentations. As known, polymeric partial prostheses constitute temporary solutions for the treatment of edentaton states. Nowadays, this treatment includes traditional acrylic prostheses, as well as elastic prostheses made of different material systems. Such a system is Valplast, which uses a polyamide for the realization of the prosthetic base. Both the rigid and the elastic materials are nowadays under debate, different – positive or negative – opinions being uttered in relation with their utilization. The scope of the present study is to support the intensive application of the elastic materials. Extremely important is the identification of the intrinsic qualities of the materials influencing the behaviour in the oral cavity, especially the effects of pressure upon the biological structures of the prosthetic field.

  8. The dppBCDF gene cluster of Haemophilus influenzae: Role in heme utilization

    Directory of Open Access Journals (Sweden)

    Morton Daniel J

    2009-08-01

    Full Text Available Abstract Background Haemophilus influenzae requires a porphyrin source for aerobic growth and possesses multiple mechanisms to obtain this essential nutrient. This porphyrin requirement may be satisfied by either heme alone, or protoporphyrin IX in the presence of an iron source. One protein involved in heme acquisition by H. influenzae is the periplasmic heme binding protein HbpA. HbpA exhibits significant homology to the dipeptide and heme binding protein DppA of Escherichia coli. DppA is a component of the DppABCDF peptide-heme permease of E. coli. H. influenzae homologs of dppBCDF are located in the genome at a point distant from hbpA. The object of this study was to investigate the potential role of the H. influenzae dppBCDF locus in heme utilization. Findings An insertional mutation in dppC was constructed and the impact of the mutation on the utilization of both free heme and various proteinaceous heme sources as well as utilization of protoporphyrin IX was determined in growth curve studies. The dppC insertion mutant strain was significantly impacted in utilization of all tested heme sources and protoporphyin IX. Complementation of the dppC mutation with an intact dppCBDF gene cluster in trans corrected the growth defects seen in the dppC mutant strain. Conclusion The dppCBDF gene cluster constitutes part of the periplasmic heme-acquisition systems of H. influenzae.

  9. Alteration by irradiation and storage at amount of heme iron in poultry meat; Alteracoes provocadas pela irradiacao e armazenamento nos teores de ferro heme em carne de frango

    Energy Technology Data Exchange (ETDEWEB)

    Souza, Adriana Regia Marques de; Arthur, Valter Arthur [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Irradiacao de Alimentos e Radioentomologia; Canniatti-Brazaca, Solange Guidolin [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Agroindustria, Alimentos e Nutricao]. E-mail: sgcbraza@esalq.usp.br

    2007-04-15

    Studies of irradiation and storage effects in chicken were carried out to discover the influence in iron heme, non-heme amount, color and total pigments. Chicken thighs and chicken breast were studied. These were irradiated to 0, 1 and 2 kGy stored by 14 days to 4 deg C in refrigerator. Determining the heme content and non-heme of meat was done using the colorimeter method and the Ferrozine reagent. The values of iron heme were influenced both by the irradiation and the storage, reducing the amount throughout the course of time. The iron non-heme was also influenced by the doses and the storage time, however the values increased throughout the course of time, because of the conversion of iron heme in non-heme. The color did not show that it was influenced by the studied doses, except for the storage, and the total number of pigments was affected by the irradiation and the time, reducing the values with the increase of storage. Irradiation was shown to be a good method to conserve iron. (author)

  10. Increased Heme Levels in the Heart Lead to Exacerbated Ischemic Injury.

    Science.gov (United States)

    Sawicki, Konrad Teodor; Shang, Meng; Wu, Rongxue; Chang, Hsiang-Chun; Khechaduri, Arineh; Sato, Tatsuya; Kamide, Christine; Liu, Ting; Naga Prasad, Sathyamangla V; Ardehali, Hossein

    2015-07-31

    Heme is an essential iron-containing molecule for cardiovascular physiology, but in excess it may increase oxidative stress. Failing human hearts have increased heme levels, with upregulation of the rate-limiting enzyme in heme synthesis, δ-aminolevulinic acid synthase 2 (ALAS2), which is normally not expressed in cardiomyocytes. We hypothesized that increased heme accumulation (through cardiac overexpression of ALAS2) leads to increased oxidative stress and cell death in the heart. We first showed that ALAS2 and heme levels are increased in the hearts of mice subjected to coronary ligation. To determine the causative role of increased heme in the development of heart failure, we generated transgenic mice with cardiac-specific overexpression of ALAS2. While ALAS2 transgenic mice have normal cardiac function at baseline, their hearts display increased heme content, higher oxidative stress, exacerbated cell death, and worsened cardiac function after coronary ligation compared to nontransgenic littermates. We confirmed in cultured cardiomyoblasts that the increased oxidative stress and cell death observed with ALAS2 overexpression is mediated by increased heme accumulation. Furthermore, knockdown of ALAS2 in cultured cardiomyoblasts exposed to hypoxia reversed the increases in heme content and cell death. Administration of the mitochondrial antioxidant MitoTempo to ALAS2-overexpressing cardiomyoblasts normalized the elevated oxidative stress and cell death levels to baseline, indicating that the effects of increased ALAS2 and heme are through elevated mitochondrial oxidative stress. The clinical relevance of these findings was supported by the finding of increased ALAS2 induction and heme accumulation in failing human hearts from patients with ischemic cardiomyopathy compared to nonischemic cardiomyopathy. Heme accumulation is detrimental to cardiac function under ischemic conditions, and reducing heme in the heart may be a novel approach for protection against the

  11. Redox and light control the heme-sensing activity of AppA.

    Science.gov (United States)

    Yin, Liang; Dragnea, Vladimira; Feldman, George; Hammad, Loubna A; Karty, Jonathan A; Dann, Charles E; Bauer, Carl E

    2013-08-27

    The DNA binding activity of the photosystem-specific repressor PpsR is known to be repressed by the antirepressor AppA. AppA contains a blue-light-absorbing BLUF domain and a heme-binding SCHIC domain that controls the interaction of AppA with PpsR in response to light and heme availability. In this study, we have solved the structure of the SCHIC domain and identified the histidine residue that is critical for heme binding. We also demonstrate that dark-adapted AppA binds heme better than light-excited AppA does and that heme bound to the SCHIC domain significantly reduces the length of the BLUF photocycle. We further show that heme binding to the SCHIC domain is affected by the redox state of a disulfide bridge located in the Cys-rich carboxyl-terminal region. These results demonstrate that light, redox, and heme are integrated inputs that control AppA's ability to disrupt the DNA binding activity of PpsR. Photosynthetic bacteria must coordinate synthesis of the tetrapyrroles cobalamin, heme, and bacteriochlorophyll, as overproduction of the latter two is toxic to cells. A key regulator controlling tetrapyrrole biosynthesis is PpsR, and the activity of PpsR is controlled by the heme-binding and light-regulated antirepressor AppA. We show that AppA binds heme only under dark conditions and that heme binding significantly affects the length of the AppA photocycle. Since AppA interacts with PpsR only in the dark, bound heme thus stimulates the antirepressor activity of PpsR. This causes the redirection of tetrapyrrole biosynthesis away from heme into the bacteriochlorophyll branch.

  12. Viability of Controlling Prosthetic Hand Utilizing Electroencephalograph (EEG) Dataset Signal

    Science.gov (United States)

    Miskon, Azizi; A/L Thanakodi, Suresh; Raihan Mazlan, Mohd; Mohd Haziq Azhar, Satria; Nooraya Mohd Tawil, Siti

    2016-11-01

    This project presents the development of an artificial hand controlled by Electroencephalograph (EEG) signal datasets for the prosthetic application. The EEG signal datasets were used as to improvise the way to control the prosthetic hand compared to the Electromyograph (EMG). The EMG has disadvantages to a person, who has not used the muscle for a long time and also to person with degenerative issues due to age factor. Thus, the EEG datasets found to be an alternative for EMG. The datasets used in this work were taken from Brain Computer Interface (BCI) Project. The datasets were already classified for open, close and combined movement operations. It served the purpose as an input to control the prosthetic hand by using an Interface system between Microsoft Visual Studio and Arduino. The obtained results reveal the prosthetic hand to be more efficient and faster in response to the EEG datasets with an additional LiPo (Lithium Polymer) battery attached to the prosthetic. Some limitations were also identified in terms of the hand movements, weight of the prosthetic, and the suggestions to improve were concluded in this paper. Overall, the objective of this paper were achieved when the prosthetic hand found to be feasible in operation utilizing the EEG datasets.

  13. Predicting prosthetic prescription after major lower-limb amputation.

    Science.gov (United States)

    Resnik, Linda; Borgia, Matthew

    2015-01-01

    We describe prosthetic limb prescription in the first year following lower-limb amputation and examine the relationship between amputation level, geographic region, and prosthetic prescription. We analyzed 2005 to 2010 Department of Veterans Affairs (VA) Inpatient and Medical Encounters SAS data sets, Vital Status death data, and National Prosthetic Patient Database data for 9,994 Veterans who underwent lower-limb amputation at a VA hospital. Descriptive statistics and bivariates were examined. Cox proportional hazard models identified factors associated with prosthetic prescription. Analyses showed that amputation level was associated with prosthetic prescription. The hazard ratios (HRs) were 1.41 for ankle amputation and 0.46 for transfemoral amputation compared with transtibial amputation. HRs for geographic region were Northeast = 1.49, Upper Midwest = 1.26, and West = 1.39 compared with the South (p prosthetic prescription. Being married was positively associated. After adjusting for patient characteristics, people with ankle amputation were most likely to be prescribed a prosthesis and people with transfemoral amputation were least likely. Geographic variation in prosthetic prescription exists in the VA and further research is needed to explain why.

  14. Performance of conventional and X2® prosthetic knees during slope descent.

    Science.gov (United States)

    Bell, Elizabeth M; Pruziner, Alison L; Wilken, Jason M; Wolf, Erik J

    2016-03-01

    Individuals with transfemoral amputation often have difficulty descending sloped surfaces due to increased lower extremity range of motion and torque requirements. The X2®, a new microprocessor-controlled prosthetic knee, claims to improve gait over sloped terrain. The aim of this study was to evaluate how experienced prosthesis users descended a sloped surface using the X2®, compared to a conventional knee, either mechanical (MECH) or microprocessor (MP). Descent technique and biomechanics were assessed in 21 service members with unilateral transfemoral amputation as they descended an instrumented 10° slope at a self-selected walking velocity. Use of the X2® in the MECH group resulted in greater hill assessment scores (8.5 to 11.0, P=0.026), due primarily to decreased reliance on handrail use. The use of the X2® in the MP group increased prosthetic knee flexion to a median of 6.4° at initial contact (P=0.002) and 73.7° in swing (P=0.005), contributing to longer prosthetic limb steps (P=0.024) and increased self-selected velocity (P=0.041). Additionally, the use of the X2® in the MP group increased prosthetic limb impact peaks (11.6N/kg, P=0.004), improving impact peak symmetry to -1.3% (P=0.004). Decreased reliance on handrail use as MECH users descended in the X2® indicate improved function and perhaps greater confidence in the device. Additional biomechanical improvements for existing MP users suggest potential longer-term benefits with regard to intact limb health and overuse injuries. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Perceived self-efficacy and specific self-reported outcomes in persons with lower-limb amputation using a non-microprocessor-controlled versus a microprocessor-controlled prosthetic knee.

    Science.gov (United States)

    Möller, Saffran; Hagberg, Kerstin; Samulesson, Kersti; Ramstrand, Nerrolyn

    2018-04-01

    To measure self-efficacy in a group of individuals who have undergone a lower-limb amputation and investigate the relationship between self-efficacy and prosthetic-specific outcomes including prosthetic use, mobility, amputation-related problems and global health. A second purpose was to examine if differences exist in outcomes based upon the type of prosthetic knee unit being used. Cross-sectional study using the General Self-Efficacy (GSE) Scale and the Questionnaire for Persons with a Transfemoral Amputation (Q-TFA). Forty-two individuals participated in the study. Twenty-three used a non-microprocessor-controlled prosthetic knee joint (non-MPK) and 19 used a microprocessor-controlled prosthetic knee joint (MPK). The study sample had quite high GSE scores (32/40). GSE scores were significantly correlated to the Q-TFA prosthetic use, mobility and problem scores. High GSE scores were related to higher levels of prosthetic use, mobility, global scores and negatively related to problem score. No significant difference was observed between individuals using a non-MPK versus MPK joints. Individuals with high self-efficacy used their prosthesis to a higher degree and high self-efficacy was related to higher level of mobility, global scores and fewer problems related to the amputation in individuals who have undergone a lower-limb amputation and were using a non-MPK or MPK knee. Implications for rehabilitation Perceived self-efficacy has has been shown to be related to quality of life, prosthetic mobility and capability as well as social activities in daily life. Prosthetic rehabilitation is primary focusing on physical improvement rather than psychological interventions. More attention should be directed towards the relationship between self-efficacy and prosthetic related outcomes during prosthetic rehabilitation after a lower-limb amputation.

  16. Modulation of function in a minimalist heme-binding membrane protein.

    Science.gov (United States)

    Shinde, Sandip; Cordova, Jeanine M; Woodrum, Brian W; Ghirlanda, Giovanna

    2012-04-01

    De novo designed heme-binding proteins have been used successfully to recapitulate features of natural hemoproteins. This approach has now been extended to membrane-soluble model proteins. Our group designed a functional hemoprotein, ME1, by engineering a bishistidine binding site into a natural membrane protein, glycophorin A (Cordova et al. in J Am Chem Soc 129:512-518, 2007). ME1 binds iron(III) protoporphyrin IX with submicromolar affinity, has a redox potential of -128 mV, and displays peroxidase activity. Here, we show the effect of aromatic residues in modulating the redox potential in the context of a membrane-soluble model system. We designed aromatic interactions with the heme through a single-point mutant, G25F, in which a phenylalanine is designed to dock against the porphyrin ring. This mutation results in roughly tenfold tighter binding to iron(III) protoporphyrin IX (K(d,app) = 6.5 × 10(-8) M), and lowers the redox potential of the cofactor to -172 mV. This work demonstrates that specific design features aimed at controlling the properties of bound cofactors can be introduced in a minimalist membrane hemoprotein model. The ability to modulate the redox potential of cofactors embedded in artificial membrane proteins is crucial for the design of electron transfer chains across membranes in functional photosynthetic devices. © SBIC 2012

  17. Acute stress-induced antinociception is cGMP-dependent but heme oxygenase-independent

    Energy Technology Data Exchange (ETDEWEB)

    Carvalho-Costa, P.G. [Programa de Graduação em Psicobiologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Branco, L.G.S. [Departamento de Morfologia, Fisiologia e Patologia Básica, Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Leite-Panissi, C.R.A. [Programa de Graduação em Psicobiologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Departamento de Morfologia, Fisiologia e Patologia Básica, Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil)

    2014-09-19

    Endogenous carbon monoxide (CO), which is produced by the enzyme heme oxygenase (HO), participates as a neuromodulator in physiological processes such as thermoregulation and nociception by stimulating the formation of 3′,5′-cyclic guanosine monophosphate (cGMP). In particular, the acute physical restraint-induced fever of rats can be blocked by inhibiting the enzyme HO. A previous study reported that the HO-CO-cGMP pathway plays a key phasic antinociceptive role in modulating noninflammatory acute pain. Thus, this study evaluated the involvement of the HO-CO-cGMP pathway in antinociception induced by acute stress in male Wistar rats (250-300 g; n=8/group) using the analgesia index (AI) in the tail flick test. The results showed that antinociception induced by acute stress was not dependent on the HO-CO-cGMP pathway, as neither treatment with the HO inhibitor ZnDBPG nor heme-lysinate altered the AI. However, antinociception was dependent on cGMP activity because pretreatment with the guanylate cyclase inhibitor 1H-[1,2,4] oxadiazolo [4,3-a] quinoxaline-1-one (ODQ) blocked the increase in the AI induced by acute stress.

  18. Bile fluorescence, heme oxygenase induction, and increased biliverdin excretion by mixtures of environmental toxicants

    Energy Technology Data Exchange (ETDEWEB)

    Hurk, Peter van den [Department of Biological Sciences, Institute of Environmental Toxicology, Clemson University, Pendleton, SC (United States)]. E-mail: pvdhurk@clemson.edu

    2006-05-01

    The measurement of bile fluorescence has become a popular biomarker to demonstrate the exposure of fish to polycyclic aromatic hydrocarbons. Conflicting data has been published on how to normalize bile fluorescence. To investigate if normalization to biliverdin is a suitable method, experiments were performed to study the mechanisms related to biliverdin excretion in fish. In two separate experiments channel catfish (Ictalurus punctatus) were dosed with mixtures of benzo[a]pyrene with cadmium, chlorinated phenols or borneol. The results showed linear relationships between bile protein and biliverdin for each treatment group, but the slope of this relationship was significantly increased when fish received more chemical stress. Thus, under increasing toxicant stress, more biliverdin was excreted per amount of protein. To investigate if the increased biliverdin excretion was related to increased heme degradation, enzymatic activity of heme oxygenase (HO) was measured in liver homogenates of the dosed fish. The fish dosed with chemical mixtures had a significantly higher HO activity than the control fish, and in both experiments a significant correlation was observed between HO activity and biliverdin concentration in the bile. It is concluded that mixtures of environmental pollutants can induce HO activity and that this chemical stress leads to increased biliverdin excretion. The elucidation of this mechanistic pathway warrants that bile fluorescence should not be expressed per biliverdin absorption, and that expression per bile protein would be a more reliable method.

  19. Bile fluorescence, heme oxygenase induction, and increased biliverdin excretion by mixtures of environmental toxicants.

    Science.gov (United States)

    van den Hurk, Peter

    2006-05-01

    The measurement of bile fluorescence has become a popular biomarker to demonstrate the exposure of fish to polycyclic aromatic hydrocarbons. Conflicting data has been published on how to normalize bile fluorescence. To investigate if normalization to biliverdin is a suitable method, experiments were performed to study the mechanisms related to biliverdin excretion in fish. In two separate experiments channel catfish (Ictalurus punctatus) were dosed with mixtures of benzo[a]pyrene with cadmium, chlorinated phenols or borneol. The results showed linear relationships between bile protein and biliverdin for each treatment group, but the slope of this relationship was significantly increased when fish received more chemical stress. Thus, under increasing toxicant stress, more biliverdin was excreted per amount of protein. To investigate if the increased biliverdin excretion was related to increased heme degradation, enzymatic activity of heme oxygenase (HO) was measured in liver homogenates of the dosed fish. The fish dosed with chemical mixtures had a significantly higher HO activity than the control fish, and in both experiments a significant correlation was observed between HO activity and biliverdin concentration in the bile. It is concluded that mixtures of environmental pollutants can induce HO activity and that this chemical stress leads to increased biliverdin excretion. The elucidation of this mechanistic pathway warrants that bile fluorescence should not be expressed per biliverdin absorption, and that expression per bile protein would be a more reliable method.

  20. A Miniature Force Sensor for Prosthetic Hands

    Science.gov (United States)

    Platt, Robert; Chu, Mars; Diftler, Myron; Martin, Toby; Valvo, Michael

    2006-01-01

    Tactile sensing is an important part of the development of new prosthetic hands. A number of approaches to establishing an afferent pathway back to the patient for tactile information are becoming available including tactors and direct stimulation of the afferent nerves. Tactile information can also be used by low-level control systems that perform simple tasks for the patient such as establishing a stable grasp and maintaining the grasping forces needed to hold an object. This abstract reports on the design of a small fingertip load cell based on semi-conductor strain gauges. Since this load cell is so small (measuring only 8.5mm in diameter and 6.25 mm in height), it easily fits into the tip of an anthropomorphic mechatronic hand. This load cell is tested by comparing a time series of force and moment data with reference data acquired from a much larger high-precision commercial load cell.

  1. [Improving the speech with a prosthetic construction].

    Science.gov (United States)

    Stalpers, M J; Engelen, M; van der Stappen, J A A M; Weijs, W L J; Takes, R P; van Heumen, C C M

    2016-03-01

    A 12-year-old boy had problems with his speech due to a defect in the soft palate. This defect was caused by the surgical removal of a synovial sarcoma. Testing with a nasometer revealed hypernasality above normal values. Given the size and severity of the defect in the soft palate, the possibility of improving the speech with speech therapy was limited. At a centre for special dentistry an attempt was made with a prosthetic construction to improve the performance of the palate and, in that way, the speech. This construction consisted of a denture with an obturator attached to it. With it, an effective closure of the palate could be achieved. New measurements with acoustic nasometry showed scores within the normal values. The nasality in the speech largely disappeared. The obturator is an effective and relatively easy solution for palatal insufficiency resulting from surgical resection. Intrusive reconstructive surgery can be avoided in this way.

  2. Construct Validity of the Prosthetic Limb Users Survey of Mobility (PLUS-M) in Adults With Lower Limb Amputation.

    Science.gov (United States)

    Hafner, Brian J; Gaunaurd, Ignacio A; Morgan, Sara J; Amtmann, Dagmar; Salem, Rana; Gailey, Robert S

    2017-02-01

    To assess construct validity of the Prosthetic Limb Users Survey of Mobility (PLUS-M), a self-report mobility measure for people with lower limb amputation (LLA). Cross-sectional study. Private prosthetic clinics (n=37). Current lower limb prosthesis users (N=199; mean age ± SD, 55.4±14.3y; 71.4% men) were assessed before receiving a replacement prosthesis, prosthetic socket, and/or prosthetic knee. Not applicable. Convergent construct validity was examined using correlations between participants' PLUS-M T-scores and measures of physical function, mobility, and balance, including the Amputee Mobility Predictor (AMP), timed Up and Go (TUG), Patient-Reported Outcomes Measurement Information System-Physical Function (PROMIS-PF), Prosthesis Evaluation Questionnaire-Mobility Subscale (PEQ-MS), and Activities-specific Balance Confidence (ABC) Scale. Known-groups construct validity was evaluated by comparing differences in PLUS-M T-scores among participants grouped by Medicare Functional Classification Level (MFCL). PLUS-M T-scores demonstrated a moderate positive relationship with AMP scores (ρ=.54, Pprosthetic mobility. Correlations between PLUS-M and measures of physical function, mobility, and balance indicate convergent construct validity. Similarly, significant differences in PLUS-M T-scores across MFCL groups provide evidence of known-groups construct validity. In summary, evidence indicates that PLUS-M has good construct validity among people with LLA. Copyright © 2016 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.

  3. SpyB, a Small Heme-Binding Protein, Affects the Composition of the Cell Wall in Streptococcus pyogenes.

    Science.gov (United States)

    Edgar, Rebecca J; Chen, Jing; Kant, Sashi; Rechkina, Elena; Rush, Jeffrey S; Forsberg, Lennart S; Jaehrig, Bernhard; Azadi, Parastoo; Tchesnokova, Veronika; Sokurenko, Evgeni V; Zhu, Haining; Korotkov, Konstantin V; Pancholi, Vijay; Korotkova, Natalia

    2016-01-01

    Streptococcus pyogenes (Group A Streptococcus or GAS) is a hemolytic human pathogen associated with a wide variety of infections ranging from minor skin and throat infections to life-threatening invasive diseases. The cell wall of GAS consists of peptidoglycan sacculus decorated with a carbohydrate comprising a polyrhamnose backbone with immunodominant N-acetylglucosamine side-chains. All GAS genomes contain the spyBA operon, which encodes a 35-amino-acid membrane protein SpyB, and a membrane-bound C3-like ADP-ribosyltransferase SpyA. In this study, we addressed the function of SpyB in GAS. Phenotypic analysis of a spyB deletion mutant revealed increased bacterial aggregation, and reduced sensitivity to β-lactams of the cephalosporin class and peptidoglycan hydrolase PlyC. Glycosyl composition analysis of cell wall isolated from the spyB mutant suggested an altered carbohydrate structure compared with the wild-type strain. Furthermore, we found that SpyB associates with heme and protoporphyrin IX. Heme binding induces SpyB dimerization, which involves disulfide bond formation between the subunits. Thus, our data suggest the possibility that SpyB activity is regulated by heme.

  4. Gait analysis in lower-limb amputation and prosthetic rehabilitation.

    Science.gov (United States)

    Esquenazi, Alberto

    2014-02-01

    Gait analysis combined with sound clinical judgment plays an important role in elucidating the factors involved in the pathologic prosthetic gait and the selection and effects of available interventions to optimize it. Detailed clinical evaluation of walking contributes to the analysis of the prosthetic gait, but evaluation in the gait laboratory using kinetic and kinematic data is often necessary to quantify and identify the particular contributions of the variables impacting the gait with confidence and assess the results of such intervention. The same approach can be considered when selecting prosthetic components and assessing leg length in this patient population. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Thrombolytic therapy in prosthetic valve thrombosis during early pregnancy

    Directory of Open Access Journals (Sweden)

    B.C. Srinivas

    2012-01-01

    Full Text Available Regardless of the improvements in the design of prosthetic heart valves and the use of anticoagulation, systemic embolism and valve thrombosis remains the most dreaded complications of mechanical heart valve replacement. A course of thrombolytic therapy may be considered as a first-line therapy for prosthetic heart valve thrombosis. The safety of thrombolysis in early pregnancy is not known. We describe a primigravida with mitral valve replacement status presenting with acute prosthetic valve thrombosis and treated successfully with intravenous streptokinase.

  6. Mechanical testing of pericardium for manufacturing prosthetic heart valves.

    Science.gov (United States)

    Aguiari, Paola; Fiorese, Michele; Iop, Laura; Gerosa, Gino; Bagno, Andrea

    2016-01-01

    Mammalian pericardia are currently used for the production of percutaneous prosthetic heart valves. The characteristics of biological tissues largely influence the durability of prosthetic devices used in the percutaneous approach and in traditional surgery, too. This paper reviews methodologies employed to assess and compare mechanical properties of pericardial patches from different mammalian species in order to identify the biomaterials adequate for manufacturing prosthetic heart valves. © The Author 2015. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.

  7. Functional imaging: monitoring heme oxygenase-1 gene expression in vivo

    Science.gov (United States)

    Zhang, Weisheng; Reilly-Contag, Pamela; Stevenson, David K.; Contag, Christopher H.

    1999-07-01

    The regulation of genetic elements can be monitored in living animals using photoproteins as reporters. Heme oxygenase (HO) is the key catabolic enzyme in the heme degradation pathway. Here, HO expression serves as a model for in vivo functional imaging of transcriptional regulation of a clinically relevant gene. HO enzymatic activity is inhibited by heme analogs, metalloporphyrins, but many members of this family of compounds also activate transcription of the HO-1 promoter. The degree of transcriptional activation by twelve metalloporphyrins, differing at the central metal and porphyrin ring substituents, was evaluated in both NIH 3T3 stable lines and transgenic animals containing HO-1 promoter-luciferase gene fusions. In the correlative cell culture assays, the metalloporphyrins increased transcription form the full length HO promoter fusion to varying degrees, but none increased transcription from a truncated HO-1 promoter. These results suggested that one or both of the two distal enhancer elements located at -4 and -10 Kb upstream from transcriptional start are required for HO-1 induction by heme and its analogs. The full-length HO-1-luc fusion was then evaluated as a transgene in mice. It was possible to monitor the effects of the metalloporphyrins, SnMP and ZnPP, in living animals over time. This spatiotemporal analyses of gene expression in vivo implied that alterations in porphyrin ring substituents and the central metal may affect the extent of gene activation. These data further indicate that using photoprotein reporters, subtle differences in gene expression can be monitored in living animals.

  8. Cyanide binding to human plasma heme-hemopexin: A comparative study

    Energy Technology Data Exchange (ETDEWEB)

    Ascenzi, Paolo, E-mail: ascenzi@uniroma3.it [Laboratorio Interdipartimentale di Microscopia Elettronica, Universita Roma Tre, Roma (Italy); Istituto Nazionale di Biostrutture e Biosistemi, Roma (Italy); Leboffe, Loris [Istituto Nazionale di Biostrutture e Biosistemi, Roma (Italy); Polticelli, Fabio [Dipartimento di Biologia, Universita Roma Tre, Roma (Italy)

    2012-11-16

    Highlights: Black-Right-Pointing-Pointer Cyanide binding to ferric HHPX-heme-Fe. Black-Right-Pointing-Pointer Cyanide binding to ferrous HHPX-heme-Fe. Black-Right-Pointing-Pointer Dithionite-mediated reduction of ferric HHPX-heme-Fe-cyanide. Black-Right-Pointing-Pointer Cyanide binding to HHPX-heme-Fe is limited by ligand deprotonation. Black-Right-Pointing-Pointer Cyanide dissociation from HHPX-heme-Fe-cyanide is limited by ligand protonation. -- Abstract: Hemopexin (HPX) displays a pivotal role in heme scavenging and delivery to the liver. In turn, heme-Fe-hemopexin (HPX-heme-Fe) displays heme-based spectroscopic and reactivity properties. Here, kinetics and thermodynamics of cyanide binding to ferric and ferrous hexa-coordinate human plasma HPX-heme-Fe (HHPX-heme-Fe(III) and HHPX-heme-Fe(II), respectively), and for the dithionite-mediated reduction of the HHPX-heme-Fe(III)-cyanide complex, at pH 7.4 and 20.0 Degree-Sign C, are reported. Values of thermodynamic and kinetic parameters for cyanide binding to HHPX-heme-Fe(III) and HHPX-heme-Fe(II) are K = (4.1 {+-} 0.4) Multiplication-Sign 10{sup -6} M, k{sub on} = (6.9 {+-} 0.5) Multiplication-Sign 10{sup 1} M{sup -1} s{sup -1}, and k{sub off} = 2.8 Multiplication-Sign 10{sup -4} s{sup -1}; and H = (6 {+-} 1) Multiplication-Sign 10{sup -1} M, h{sub on} = 1.2 Multiplication-Sign 10{sup -1} M{sup -1} s{sup -1}, and h{sub off} = (7.1 {+-} 0.8) Multiplication-Sign 10{sup -2} s{sup -1}, respectively. The value of the rate constant for the dithionite-mediated reduction of the HHPX-heme-Fe(III)-cyanide complex is l = 8.9 {+-} 0.8 M{sup -1/2} s{sup -1}. HHPX-heme-Fe reactivity is modulated by proton acceptor/donor amino acid residue(s) (e.g., His236) assisting the deprotonation and protonation of the incoming and outgoing ligand, respectively.

  9. Dietary Heme Induces Gut Dysbiosis, Aggravates Colitis, and Potentiates the Development of Adenomas in Mice

    Directory of Open Access Journals (Sweden)

    Marco Constante

    2017-09-01

    Full Text Available Dietary heme can be used by colonic bacteria equipped with heme-uptake systems as a growth factor and thereby impact on the microbial community structure. The impact of heme on the gut microbiota composition may be particularly pertinent in chronic inflammation such as in inflammatory bowel disease (IBD, where a strong association with gut dysbiosis has been consistently reported. In this study we investigated the influence of dietary heme on the gut microbiota and inferred metagenomic composition, and on chemically induced colitis and colitis-associated adenoma development in mice. Using 16S rRNA gene sequencing, we found that mice fed a diet supplemented with heme significantly altered their microbiota composition, characterized by a decrease in α-diversity, a reduction of Firmicutes and an increase of Proteobacteria, particularly Enterobacteriaceae. These changes were similar to shifts seen in dextran sodium sulfate (DSS-treated mice to induce colitis. In addition, dietary heme, but not systemically delivered heme, contributed to the exacerbation of DSS-induced colitis and facilitated adenoma formation in the azoxymethane/DSS colorectal cancer (CRC mouse model. Using inferred metagenomics, we found that the microbiota alterations elicited by dietary heme resulted in non-beneficial functional shifts, which were also characteristic of DSS-induced colitis. Furthermore, a reduction in fecal butyrate levels was found in mice fed the heme supplemented diet compared to mice fed the control diet. Iron metabolism genes known to contribute to heme release from red blood cells, heme uptake, and heme exporter proteins, were significantly enriched, indicating a shift toward favoring the growth of bacteria able to uptake heme and protect against its toxicity. In conclusion, our data suggest that luminal heme, originating from dietary components or gastrointestinal bleeding in IBD and, to lesser extent in CRC, directly contributes to microbiota dysbiosis

  10. Vernal keratoconjunctivitis: culmination of management using immunosuppression, surgical and prosthetic therapy over quarter century.

    Science.gov (United States)

    Das, Shilpa; Pasari, Anand S; Sangwan, Virender S

    2016-11-23

    A 22-year-old male patient presented in 1988 with active vernal keratoconjunctivitis. He was treated with topical mast cell stabilisers and corticosteroids. Chronic inflammation despite topical treatment necessitated oral immunosuppressants. Active disease came under control with this; however, the patient gradually developed limbal stem cell deficiency. He underwent bilateral pannus resection with amniotic membrane transplantation that resulted in improved ocular surface. In 2007, patient was found to have significant bilateral posterior subcapsular cataracts and underwent bilateral cataract surgery with intraocular lens implantation with good visual outcome. In 2016, he was provided with scleral lens prosthetic device, which further improved vision. At last follow-up, more than 25 years after his initial visit, his visual acuity was 20/25 in both eyes with a stable surface. With a comprehensive approach using immunosuppression, surgical therapy and scleral lens prosthetic device, chronic vernal keratoconjunctivitis can be well managed as illustrated in this case. 2016 BMJ Publishing Group Ltd.

  11. Characterization of microbes in prosthetic joint specimens by culture-independent molecular methods

    DEFF Research Database (Denmark)

    Xu, Yijuan; Rudkjøbing, Vibeke Børsholt; Simonsen, Ole

    was to investigate the microbial diversity in surgical samples (eg. synovial fluid, periprosthetic tissue, removed prosthesis) from 22 prosthetic patients using a range of culture-independent molecular methods including broad range 16S rRNA gene PCR, cloning, phylogeny, quantitative PCR (qPCR), and fluorescence......Prosthetic joint infection (PJI) is one of the most challenging complications of joint alloplasty. Formation of biofilm is a prominent feature of PJIs and constitutes a challenge to current sampling procedures and culture practices to obtain a reliable diagnosis. The aim of the study...... in situ hybridization (FISH). Concomitant samples were cultured by standard methods. Overall, the results of culture-based and molecular methods showed concordant results for 13 patients and discrepant results for 6 patients. In the remaining cases, culture methods identified one species or a group...

  12. Microsomal prostaglandin E synthase type 2 (mPGES2) is a glutathione-dependent heme protein, and dithiothreitol dissociates the bound heme to produce active prostaglandin E2 synthase in vitro.

    Science.gov (United States)

    Takusagawa, Fusao

    2013-04-05

    An x-ray study indicated that microsomal prostaglandin E synthase type 2 (mPGES2) is a heme-bound protein and catalyzes prostaglandin (PG) H2 degradation, but not PGE2 formation (Yamada, T., and Takusagawa, F. (2007) Biochemistry 46, 8414-8424). In response to the x-ray study, Watanabe et al. claimed that mPGES2 is a heme-free protein and that both the heme-free and heme-bound proteins have PGE2 synthesis activity in the presence of dithiothreitol (Watanabe, K., Ito, S., and Yamamoto, S. (2008) Biochem. Biophys. Res. Commun. 367, 782-786). To resolve the contradictory results, the heme-binding scheme of mPGES2 was further characterized in vivo and in vitro by absorption and fluorescence spectroscopies. A substantial amount of heme-bound mPGES2 was detected in cell extracts. The heme content in mPGES2 was increased along with an increase in Fe(3+) in the culture medium. Heme-free mPGES2 was converted to the heme-bound form by mixing it with pig liver extract, indicating that mPGES2 is capable of forming a complex with heme in mammalian cells. Heme binds to mPGES2 only in the presence of glutathione. The newly determined heme dissociation constant (2.9 nM) supports strongly that mPGES2 is a heme-bound protein in vivo. The bound heme was not dissociated by oxidation by H2O2 or reduction by glutathione or 2-mercaptoethanol. However, reduction by dithiothreitol (an artificial reducing compound) induced the bound heme to dissociate from mPGES2 and released heme-free mPGES2, which exhibited PGE2 synthesis activity in vitro. Imidazole bound to mPGES2 by stacking on the bound heme and inhibited heme oxidation by H2O2 and reduction by dithiothreitol.

  13. Nitric oxide heme interactions in nitrophorin from Cimex lectularius

    Energy Technology Data Exchange (ETDEWEB)

    Christmann, R.; Auerbach, H., E-mail: auerbach@physik.uni-kl.de [University of Kaiserslautern, Department of Physics (Germany); Berry, R. E.; Walker, F. A. [The University of Arizona, Department of Chemistry and Biochemistry (United States); Schünemann, V. [University of Kaiserslautern, Department of Physics (Germany)

    2016-12-15

    The nitrophorin from the bedbug Cimex lectularius (cNP) is a nitric oxide (NO) carrying protein. Like the nitrophorins (rNPs) from the kissing bug Rhodnius prolixus, cNP forms a stable heme Fe(III)-NO complex, where the NO can be stored reversibly for a long period of time. In both cases, the NPs are found in the salivary glands of blood-sucking bugs. The insects use the nitrophorins to transport the NO to the victim’s tissues, resulting in vasodilation and reduced blood coagulation. However, the structure of cNP is significantly different to those of the rNPs from Rhodnius prolixus. Furthermore, the cNP can bind a second NO molecule to the proximal heme cysteine when present at higher concentrations. High field Mössbauer spectroscopy on {sup 57}Fe enriched cNP complexed with NO shows reduction of the heme iron and formation of a ferrous nitric oxide (Fe(II)-NO) complex. Density functional theory calculations reproduce the experimental Mössbauer parameters and confirm this observation.

  14. Effects of prosthesis use on the capability to control myoelectric robotic prosthetic hands.

    Science.gov (United States)

    Atzori, Manfredo; Hager, Anne-Gabrielle Mittaz; Elsig, Simone; Giatsidis, Giorgio; Bassetto, Franco; Muller, Henning

    2015-08-01

    The natural control of robotic prosthetic hands with non-invasive techniques is still a challenge: myoelectric prostheses currently give some control capabilities; the application of pattern recognition techniques is promising and recently started to be applied in practice but still many questions are open in the field. In particular, the effects of clinical factors on movement classification accuracy and the capability to control myoelectric prosthetic hands are analyzed in very few studies. The effect of regularly using prostheses on movement classification accuracy has been previously studied, showing differences between users of myoelectric and cosmetic prostheses. In this paper we compare users of myoelectric and body-powered prostheses and intact subjects. 36 machine-learning methods are applied on 6 amputees and 40 intact subjects performing 40 movements. Then, statistical analyses are performed in order to highlight significant differences between the groups of subjects. The statistical analyses do not show significant differences between the two groups of amputees, while significant differences are obtained between amputees and intact subjects. These results constitute new information in the field and suggest new interpretations to previous hypotheses, thus adding precious information towards natural control of robotic prosthetic hands.

  15. Dual role of the active-center cysteine in human peroxiredoxin 1: Peroxidase activity and heme binding.

    Science.gov (United States)

    Watanabe, Yuta; Ishimori, Koichiro; Uchida, Takeshi

    2017-02-12

    HBP23, a 23-kDa heme-binding protein identified in rats, is a member of the peroxiredoxin (Prx) family, the primary peroxidases involved in hydrogen peroxide catabolism. Although HBP23 has a characteristic Cys-Pro heme-binding motif, the significance of heme binding to Prx family proteins remains to be elucidated. Here, we examined the effect of heme binding to human peroxiredoxin-1 (PRX1), which has 97% amino acid identity to HBP23. PRX1 was expressed in Escherichia coli and purified to homogeneity. Spectroscopic titration demonstrated that PRX1 binds heme with a 1:1 stoichiometry and a dissociation constant of 0.17 μM. UV-vis spectra of heme-PRX1 suggested that Cys52 is the axial ligand of ferric heme. PRX1 peroxidase activity was lost upon heme binding, reflecting the fact that Cys52 is not only the heme-binding site but also the active center of peroxidase activity. Interestingly, heme binding to PRX1 caused a decrease in the toxicity and degradation of heme, significantly suppressing H2O2-dependent heme peroxidase activity and degradation of PRX1-bound heme compared with that of free hemin. By virtue of its cytosolic abundance (∼20 μM), PRX1 thus functions as a scavenger of cytosolic hemin (dual role; Cys-dependent peroxidase activity and cytosolic heme scavenger. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Heme-mediated cell activation: the inflammatory puzzle of sickle cell anemia.

    Science.gov (United States)

    Guarda, Caroline Conceição da; Santiago, Rayra Pereira; Fiuza, Luciana Magalhães; Aleluia, Milena Magalhães; Ferreira, Júnia Raquel Dutra; Figueiredo, Camylla Vilas Boas; Yahouedehou, Setondji Cocou Modeste Alexandre; Oliveira, Rodrigo Mota de; Lyra, Isa Menezes; Gonçalves, Marilda de Souza

    2017-06-01

    Hemolysis triggers the onset of several clinical manifestations of sickle cell anemia (SCA). During hemolysis, heme, which is derived from hemoglobin (Hb), accumulates due to the inability of detoxification systems to scavenge sufficiently. Heme exerts multiple harmful effects, including leukocyte activation and migration, enhanced adhesion molecule expression by endothelial cells and the production of pro-oxidant molecules. Area covered: In this review, we describe the effects of heme on leukocytes and endothelial cells, as well as the features of vascular endothelial cells related to vaso-occlusion in SCA. Expert commentary: Free Hb, heme and iron, potent cytotoxic intravascular molecules released during hemolysis, can exacerbate, modulate and maintain the inflammatory response, a main feature of SCA. Endothelial cells in the vascular environment, as well as leukocytes, can become activated via the molecular signaling effects of heme. Due to the hemolytic nature of SCA, hemolysis represents an interesting therapeutic target for heme-scavenging purposes.

  17. Impact of prosthetic material on mid- and long-term outcome of dental implants supporting single crowns and fixed partial dentures: A systematic review and meta-analysis.

    Science.gov (United States)

    Abou-Ayash, Samir; Strasding, Malin; Rücker, Gerta; Att, Wael

    2017-01-01

    The impact of prosthetic material selection on implant survival is not clear. The current criteria for choosing a prosthetic material seem to be based on clinician preferences. This systematic review aims to evaluate the impact of restorative materials on the mid- and long-term survival of implants supporting single crowns and fixed partial dentures. Hand and MEDLINE searches were performed to identify relevant literature for single crowns (SC) and fixed partial dentures (FPD). Further inclusion criteria were a mean follow-up period of at least 3 years, the inclusion of at least 10 patients in a relevant study cohort, and a clear description of prosthesis type and prosthetic material. A total of 63 studies for the SC group and 11 studies for the FPD group were included. Full arch restorations were not included. The materials utilised in the SC group were metal-ceramic (precious and non-precious), lithium-disilicate, veneered zirconia, veneered alumina, and nanoceramics. The materials used in the FPD group were metal-ceramic (precious), veneered titanium, metal-resin (precious), and veneered zirconia. No significant impact on the prosthetic material relating to mid- or long-term implant survival was identified. Furthermore, there were no statistically significant differences between the survival rates of the dental prostheses made from different materials (SC and FPD group). Single crowns made of nanoceramics showed a higher risk for decementation relative to other materials (0.80, 95% CI [0.67; 0.89]; P material selection has no influence on mid- and long-term survival of implants restored with single crowns and fixed partial dentures. Similarly, the prosthetic material seems to have no significant impact on prosthetic survival rates. Further research is required to provide more evidence regarding the impact of the prosthetic material on long-term outcome. Conflict-of-interest statement: The authors declare that they have no conflict of interest.

  18. Human and rodent amyloid-beta peptides differentially bind heme: relevance to the human susceptibility to Alzheimer's disease.

    Science.gov (United States)

    Atamna, Hani; Frey, William H; Ko, Novie

    2009-07-01

    Amyloid-beta (Abeta) peptides are implicated in the neurodegeneration of Alzheimer's disease (AD). We previously investigated the mechanism of neurotoxicity of Abeta and found that human Abeta (huAbeta) binds and depletes heme, forming an Abeta-heme complex with peroxidase activity. Rodent Abeta (roAbeta) is identical to huAbeta, except for three amino acids within the proposed heme-binding motif (Site-H). We studied and compared heme-binding between roAbeta and huAbeta. Unlike roAbeta, huAbeta binds heme tightly (K(d)=140+/-60 nM) and forms a peroxidase. The plot of bound (huAbeta-heme) vs. unbound heme fits best to a two site binding hyperbola, suggesting huAbeta possesses two heme-binding sites. Consistently, a second high affinity heme-binding site was identified in the lipophilic region (site-L) of huAbeta (K(d)=210+/-80 nM). The plot of (roAbeta-heme) vs. unbound heme, on the other hand, was different as it fits best to a sigmoidal binding curve, indicating different binding and lower affinity of roAbeta for heme (K(d)=1 microM). The effect of heme-binding to site-H on heme-binding to site-L in roAbeta and huAbeta is discussed. While both roAbeta and huAbeta form aggregates equally, rodents lack AD-like neuropathology. High huAbeta/heme ratio increases the peroxidase activity. These findings suggest that depletion of regulatory heme and formation of Abeta-heme peroxidase contribute to huAbeta's neurotoxicity in the early stages of AD. Phylogenic variations in the amino acid sequence of Abeta explain tight heme-binding to huAbeta and likely contribute to the increased human susceptibility to AD.

  19. Irradiation of bovine meat: effect of heme-iron concentration.; Irradiacao de carne bovina: efeito na concentracao de ferro heme

    Energy Technology Data Exchange (ETDEWEB)

    Mistura, Liliana Perazzini Furtado

    2002-07-01

    The irradiation is often used, nowadays, for meat conservation and it is important to know how much this process interferes with the nutritional quality of the meat. In this study round cut meat, ground and steaks (from a local supermarket) was irradiated with doses of O; 1; 2; 3; 4; 5; 7,5 and 10 kGy (JS-7500 Nordium Inc -Canada) and the interference of irradiation and the process of food preparation on heme-iron (H Fe) content was determined. Half of the sample was kept raw and the other half was grilled in a pre-warmed oven at 250 deg C for 9 min and a controlled humidity of 70%. The chemical composition, the total iron (T Fe) (EM) and the heme iron concentration were determined (Hornsey,1956) and the sensorial quality evaluated. The average T Fe concentration of raw and ground , ground and grilled, raw steaks and grilled steak meat, on dry and degreased basis was 113 mug/g, 121 mug/g , 91 mug/g and 77 mug/g; and the H Fe concentration 105 mug/g (93% of T Fe) , 88 mug/g (73% of T Fe), 90 mug/g (99% of T Fe) and 52 mug/g (68% of T Fe) respectively. Data were evaluated by ANOVA with fixed effects and multiple comparisons. The irradiation neither altered the chemical composition nor the proportion of heme iron of meat. The preparation conditions (temperature, cooking time, environment humidity, meat presentation) of the sample interfered more with the heme iron content than the irradiation. With the sensorial analysis we verified that meats irradiated with doses of 3 kGy were better evaluated in softness and succulency attributes than the others. Meat submitted to irradiation doses up to 3 kGy were accepted by the specialists' panel. (author)

  20. Altered heme catabolism by heme oxygenase-1 caused by mutations in human NADPH cytochrome P450 reductase

    Energy Technology Data Exchange (ETDEWEB)

    Pandey, Amit V., E-mail: amit@pandeylab.org [Pediatric Endocrinology, Diabetology and Metabolism, Department of Clinical Research, University of Bern, Tiefenaustrasse 120c, CH-3004 Bern (Switzerland); Flueck, Christa E.; Mullis, Primus E. [Pediatric Endocrinology, Diabetology and Metabolism, Department of Clinical Research, University of Bern, Tiefenaustrasse 120c, CH-3004 Bern (Switzerland)

    2010-09-24

    Research highlights: {yields} Mutations in POR identified from patients lead to reduced HO-1 activities. {yields} POR mutation Y181D affecting FMN binding results in total loss of HO-1 activity. {yields} POR mutations A287P, C569Y and V608F, lost 50-70% activity. {yields} Mutations in FAD binding domain, R457H, Y459H and V492E lost all HO-1 activity. {yields} POR polymorphisms P228L, R316W, G413S, A503V and G504R have normal activity. -- Abstract: Human heme oxygenase-1 (HO-1) carries out heme catabolism supported by electrons supplied from the NADPH through NADPH P450 reductase (POR, CPR). Previously we have shown that mutations in human POR cause a rare form of congenital adrenal hyperplasia. In this study, we have evaluated the effects of mutations in POR on HO-1 activity. We used purified preparations of wild type and mutant human POR and in vitro reconstitution with purified HO-1 to measure heme degradation in a coupled assay using biliverdin reductase. Here we show that mutations in POR found in patients may reduce HO-1 activity, potentially influencing heme catabolism in individuals carrying mutant POR alleles. POR mutants Y181D, A457H, Y459H, V492E and R616X had total loss of HO-1 activity, while POR mutations A287P, C569Y and V608F lost 50-70% activity. The POR variants P228L, R316W and G413S, A503V and G504R identified as polymorphs had close to WT activity. Loss of HO-1 activity may result in increased oxidative neurotoxicity, anemia, growth retardation and iron deposition. Further examination of patients affected with POR deficiency will be required to assess the metabolic effects of reduced HO-1 activity in affected individuals.

  1. Resonance Raman Spectra of Five-Coordinate Heme-Nitrosyl Cytochromes c': Effect of the Proximal Heme-NO Environment.

    Science.gov (United States)

    Servid, Amy E; McKay, Alison L; Davis, Cherry A; Garton, Elizabeth M; Manole, Andreea; Dobbin, Paul S; Hough, Michael A; Andrew, Colin R

    2015-06-02

    Five-coordinate heme nitrosyl complexes (5cNO) underpin biological heme-NO signal transduction. Bacterial cytochromes c' are some of the few structurally characterized 5cNO proteins, exhibiting a distal to proximal 5cNO transition of relevance to NO sensing. Establishing how 5cNO coordination (distal vs proximal) depends on the heme environment is important for understanding this process. Recent 5cNO crystal structures of Alcaligenes xylosoxidans cytochrome c' (AXCP) and Shewanella frigidimarina cytochrome c' (SFCP) show a basic residue (Arg124 and Lys126, respectively) near the proximal NO binding sites. Using resonance Raman (RR) spectroscopy, we show that structurally characterized 5cNO complexes of AXCP variants and SFCP exhibit a range of ν(NO) (1651-1671 cm(-1)) and ν(FeNO) (519-536 cm(-1)) vibrational frequencies, depending on the nature of the proximal heme pocket and the sample temperature. While the AXCP Arg124 residue appears to have little impact on 5cNO vibrations, the ν(NO) and ν(FeNO) frequencies of the R124K variant are consistent with (electrostatically) enhanced Fe(II) → (NO)π* backbonding. Notably, RR frequencies for SFCP and R124A AXCP are significantly displaced from the backbonding trendline, which in light of recent crystallographic data and density functional theory modeling may reflect changes in the Fe-N-O angle and/or extent of σ-donation from the NO(π*) to the Fe(II) (dz(2)) orbital. For R124A AXCP, correlation of vibrational and crystallographic data is complicated by distal and proximal 5cNO populations. Overall, this study highlights the complex structure-vibrational relationships of 5cNO proteins that allow RR spectra to distinguish 5cNO coordination in certain electrostatic and steric environments.

  2. The impact of gender, level of amputation and diabetes on prosthetic fit rates following major lower extremity amputation.

    Science.gov (United States)

    Davie-Smith, Fiona; Paul, Lorna; Nicholls, Natalie; Stuart, Wesley P; Kennon, Brian

    2017-02-01

    Diabetes mellitus is a leading cause of major lower extremity amputation. To examine the influence of gender, level of amputation and diabetes mellitus status on being fit with a prosthetic limb following lower extremity amputation for peripheral arterial disease. Retrospective analysis of the Scottish Physiotherapy Amputee Research Group dataset. Within the cohort with peripheral arterial disease ( n = 1735), 64% were men ( n = 1112) and 48% ( n = 834) had diabetes mellitus. Those with diabetes mellitus were younger than those without: mean 67.5 and 71.1 years, respectively ( p amputation:trans-femoral amputation ratio was 2.33 in those with diabetes mellitus, and 0.93 in those without. A total of 41% of those with diabetes mellitus were successfully fit with a prosthetic limb compared to 38% of those without diabetes mellitus. Male gender positively predicted fitting with a prosthetic limb at both trans-tibial amputation ( p = 0.001) and trans-femoral amputation ( p = 0.001) levels. Bilateral amputations and increasing age were negative predictors of fitting with a prosthetic limb ( p amputation level ( p amputation was at trans-femoral amputation level. Of those with lower extremity amputation as a result of peripheral arterial disease, those with diabetes mellitus were younger, and more had trans-tibial amputation. Although both age and amputation level are good predictors of fitting with a prosthetic limb, successful limb fit rates were no better than those without diabetes mellitus. Clinical relevance This is of clinical relevance to those who are involved in the decision-making process of prosthetic fitting following major amputation for dysvascular and diabetes aetiologies.

  3. [A new implant system for orbital prosthetic rehabilitation: "epiplating mono"].

    Science.gov (United States)

    Schneider, M; Federspil, P A; Neumann, A; Schick, B

    2014-06-01

    A New Implant System for Orbital Prosthetic Rehabilitation: "Epiplating Mono" Prosthetic or episthetic rehabilitation of ear, eye and nose are currently most common performed using magnetic fixation. While at the beginning single implants have been used, now-a-days a more extended approach with plate fixation are recommended to enhance the stability of the anchored magnets. A newly designed implant system epiplating mono is presented that combines the structure of a single implant with additional fixation elements. In a pilot study this new implant system was used in 4 patients for prosthetic orbital rehabilitation. Further experiences with this new implant system are required necessitating long-term experiences of implant stability to define the value of the presented epiplating mono system for prosthetic rehabilitation. © Georg Thieme Verlag KG Stuttgart · New York.

  4. Innovations in prosthetic interfaces for the upper extremity.

    Science.gov (United States)

    Kung, Theodore A; Bueno, Reuben A; Alkhalefah, Ghadah K; Langhals, Nicholas B; Urbanchek, Melanie G; Cederna, Paul S

    2013-12-01

    Advancements in modern robotic technology have led to the development of highly sophisticated upper extremity prosthetic limbs. High-fidelity volitional control of these devices is dependent on the critical interface between the patient and the mechanical prosthesis. Recent innovations in prosthetic interfaces have focused on several control strategies. Targeted muscle reinnervation is currently the most immediately applicable prosthetic control strategy and is particularly indicated in proximal upper extremity amputations. Investigation into various brain interfaces has allowed acquisition of neuroelectric signals directly or indirectly from the central nervous system for prosthetic control. Peripheral nerve interfaces permit signal transduction from both motor and sensory nerves with a higher degree of selectivity. This article reviews the current developments in each of these interface systems and discusses the potential of these approaches to facilitate motor control and sensory feedback in upper extremity neuroprosthetic devices.

  5. Esthetics with prosthetics in case of maxillary canine transposition: A ...

    African Journals Online (AJOL)

    2011-03-16

    .app.medknow. For suggestions and comments do write back to us. Announcement. How to cite this article: Yadav S, Sheorain AK, Madan N, Bajaj P. Esthetics with prosthetics in case of maxillary canine transposition: A ...

  6. Fixed prosthetic treatment in patients with cleft lip and palate

    Directory of Open Access Journals (Sweden)

    Bajevska Jagoda

    2017-01-01

    Full Text Available Introduction. The prosthetic treatment of patients with cleft palate includes various treatment options such as fixed partial dentures, removable partial prosthesis, etc. The type of prosthetic appliance is determined by the oral health of each individual and the circumstances. We presented three adult patients with the cleft lip and palate subjected to prosthetic treatment. Case report. From the possible prosthetic solutions according to the conditions in the oral cavity and the circumstances, fixed partial dentures veneered with composite or ceramic were chosen. A proper relationship between the teeth was reached with the fixed partial dentures, and function established, the phonetics improved and satisfying aesthetics effect accomplished improving the profile appearance of the patient’s face. Plastic surgery of the nose was performed after that. Conclusion. Multidisclipinary treatment is necessary for favourable long-term outcome in cleft lip and palate patients.

  7. Prosthetic motility in pegged versus unpegged integrated porous orbital implants.

    Science.gov (United States)

    Guillinta, Paulo; Vasani, Sunil N; Granet, David B; Kikkawa, Don O

    2003-03-01

    To objectively measure and compare prosthetic motility in pegged versus unpegged orbital implants and to determine subjective patient assessment of motility after the pegging procedure. A prospective case series of 10 patients with integrated porous orbital implants, who had secondary motility peg placement procedure, were studied. Infrared oculography was used to quantitatively assess pegged and unpegged prosthetic eye motility in horizontal and vertical excursions. For horizontal excursions, prosthetic motility in unpegged implants retained an average of 49.6% of measured motility of the contralateral normal eye, which increased to 86.5% with peg placement (Ppeg placement (P>0.3). Nine of 10 patients judged their motility as "significantly improved," and 1 patient gave a rating of "some improvement" after peg placement. Four of 10 patients had granulomas around the peg sites. Objective assessment of prosthetic motility shows a significant increase in horizontal gaze after motility peg placement.

  8. Antithrombotic Therapy in Patients with Prosthetic Heart Valves

    Directory of Open Access Journals (Sweden)

    Mohamed HA

    2009-01-01

    Full Text Available Patients with mechanical valve prostheses require a lifelong anticoagulant treatment. The combined use of Warfarin and low-dose aspirin appears to reduce the risk of valve thrombosis and systemic embolism at a low risk of bleeding. The management of women with prosthetic heart valves during pregnancy poses a particular challenge, as there are no available controlled clinical trials to provide guidelines for effective antithrombotic therapy. Oral anticoagulants, such as Warfarin, cause foetal embryopathy; unfractionated heparin and low-molecular-weight heparin have been reported to be ineffective in preventing thromboembolic complications.This article discusses the available data and the most recent guidelines in the antithrombotic management of patients with prosthetic valves, and antithrombotic therapy in various clinical situations such as pregnant women with prosthetic heart valves, and patients with prosthetic heart valves undergoing noncardiac surgery.

  9. Adipose-Derived Stem Cell Modulation of T-Cell Regulation Correlates with Heme Oxgenase-1 Pathway Changes.

    Science.gov (United States)

    Chien, Ching-Ming; Chen, Yung-Wei; Chen, Chien-Chang; Wu, Yi-Chia; Huang, Shu-Hung; Lee, Su-Shin; Lai, Cheng-Sheng; Lin, Sin-Daw; Wang, Ching-Jen; Kuo, Yur-Ren

    2016-11-01

    The authors' previous proteome study revealed that haptoglobin was involved in adipose-derived stem cell modulation of allotransplant survival and T-cell regulation to induce immune tolerance. This study investigated whether adipose-derived stem cells could modulate T-cell regulation through haptoglobin and the downstream heme oxgenase-1 pathway in vitro. Splenocytes were isolated from Lewis rat spleens and then CD3 T cells were purified using anti-CD3 beads. Adipose-derived stem cells were harvested from Lewis rats and co-cultured with the T cells. After Transwell co-culture at different periods, the authors analyzed cell proliferation with a bromodeoxyuridine assay. Cell extractions and culture supernatants were collected for further analysis. Heme oxgenase-1 and related protein expression levels from the adipose-derived stem cells and T cells were detected using Western blotting. The related cytokine expression levels were analyzed with enzyme-linked immunosorbent assay kits. Flow cytometry was used to detect the regulatory T-cell proportion. The adipose-derived stem cells significantly suppressed T-cell proliferation. The regulatory T-cell percentages were significantly increased in the adipose-derived stem cells that were co-cultured with T cells compared with T cells alone without adipose-derived stem cell co-culture. Heme oxgenase-1 expression in concanavalin A-stimulated T cells that were co-cultured with adipose-derived stem cells revealed a significant increase compared with concanavalin A-stimulated T cells alone. Cytokine assays of the culture supernatants revealed that transforming growth factor-β and interleukin-10 were significantly increased and interferon-γ was statistically decreased in the adipose-derived stem cell-co-cultured T-cell group compared with other groups; however, blockade with a heme oxgenase-1 inhibitor (zinc protoporphyrin IX) protected against these changes. Adipose-derived stem cells modulate T-cell proliferation and enhance

  10. Adenoviral transfer of the heme oxygenase-1 gene protects striatal astrocytes from heme-mediated oxidative injury.

    Science.gov (United States)

    Teng, Zhi-Ping; Chen, Jing; Chau, Lee-Young; Galunic, Nicholas; Regan, Raymond F

    2004-11-01

    Heme oxygenase-1 (HO-1) is induced in the CNS after hemorrhage, and may have an effect on injury to surrounding tissue. Hemin, the preferred substrate of HO, is a neurotoxin that is present in intracranial hematomas. In a prior study, we observed that HO inhibitors increased the vulnerability of cultured cortical astrocytes to heme-mediated oxidative injury. To investigate the effect of HO more specifically, we used an adenoviral vector encoding the human HO-1 gene to specifically increase HO-1 expression. Incubation with 100 MOI of the HO-1 adenovirus (Adv-HHO-1) for 24 h increased both HO-1 protein and HO activity; a control adenovirus lacking the HO-1 gene had no effect. Using a DNA probe that was specific for human HO-1, 80.5 +/- 7.2% of astrocytes were observed to be infected by in situ hybridization. The cell death produced by 30-60 microM hemin was significantly reduced by pretreatment with 100 MOI Adv-HHO-1, as assessed by LDH release, propidium iodide exclusion, and MTT reduction assay. The threefold increase in cell protein oxidation produced by hemin was also attenuated in cultures pretreated with Adv-HHO-1. These results support the hypothesis that HO-1 protects astrocytes from heme-mediated oxidative injury. Specifically increasing astrocytic HO-1 by gene transfer may have a beneficial effect on hemorrhagic CNS injury.

  11. Heme, an Essential Nutrient from Dietary Proteins, Critically Impacts Diverse Physiological and Pathological Processes

    Directory of Open Access Journals (Sweden)

    Jagmohan Hooda

    2014-03-01

    Full Text Available Heme constitutes 95% of functional iron in the human body, as well as two-thirds of the average person’s iron intake in developed countries. Hence, a wide range of epidemiological studies have focused on examining the association of dietary heme intake, mainly from red meat, with the risks of common diseases. High heme intake is associated with increased risk of several cancers, including colorectal cancer, pancreatic cancer and lung cancer. Likewise, the evidence for increased risks of type-2 diabetes and coronary heart disease associated with high heme intake is compelling. Furthermore, recent comparative metabolic and molecular studies of lung cancer cells showed that cancer cells require increased intracellular heme biosynthesis and uptake to meet the increased demand for oxygen-utilizing hemoproteins. Increased levels of hemoproteins in turn lead to intensified oxygen consumption and cellular energy generation, thereby fueling cancer cell progression. Together, both epidemiological and molecular studies support the idea that heme positively impacts cancer progression. However, it is also worth noting that heme deficiency can cause serious diseases in humans, such as anemia, porphyrias, and Alzheimer’s disease. This review attempts to summarize the latest literature in understanding the role of dietary heme intake and heme function in diverse diseases.

  12. Grasping Pattern Recognition and Grasping Force Estimation For Prosthetic Hands

    OpenAIRE

    Zhang Bing-Ke; Zhong Guo-Liang; Deng Hua

    2016-01-01

    Human’s movement can be decoded by surface electromyography (EMG), and the prosthetic hand can be controlled freely through EMG signal. This paper proposes a grasping pattern and force synchronized decoding method for prosthetic hands. Considering pattern recognition and force estimation simultaneously, this paper analyzes whether different muscle contraction levels affect pattern recognition and whether different grasping modes have impact on force estimation, then proposes two schemes to co...

  13. 24 DOF EMG controlled hybrid actuated prosthetic hand.

    Science.gov (United States)

    Atasoy, A; Kaya, E; Toptas, E; Kuchimov, S; Kaplanoglu, E; Ozkan, M

    2016-08-01

    A complete mechanical design concept of an electromyogram (EMG) controlled hybrid prosthetic hand, with 24 degree of freedom (DOF) anthropomorphic structure is presented. Brushless DC motors along with Shape Memory Alloy (SMA) actuators are used to achieve dexterous functionality. An 8 channel EMG is used for detecting 7 basic hand gestures for control purposes. The prosthetic hand will be integrated with the Neural Network (NNE) based controller in the next phase of the study.

  14. Possibilities of prosthetic upper limb fitting in cycling

    OpenAIRE

    Doležalová, Hana

    2011-01-01

    Bachelor thesis give an overview of possible solutions in upper limbs prosthetic fitting which is suitable for cycling. And provide enough information on modifications that should be performed on a bicycle so that it can be used by humans with upper extremity prostheses. It can be an essential guide for anyone looking for a solution that would allow a person with an amputated upper limb again sit on the bike. Keywords: amputation, upper limb prosthesis, prosthetic fitting, cycling

  15. Calcium-Dependent Conformation of a Heme and Fingerprint Peptide of the Di-Heme Cytochrome c Peroxidase from Paracoccus Pantotrophus

    Energy Technology Data Exchange (ETDEWEB)

    PAULETA,SOFIA R.; LU,YI; GOODHEW,CELIA F.; MOURA,ISABEL; PETTIGREW,GRAHAM W.; SHELNUTT,JOHN A.

    2000-12-18

    The structural changes in the heme macrocycle and substituents caused by binding of Ca{sup 2+} to the diheme cytochrome c peroxidase from Paracoccuspantotrophus were clarified by resonance Raman spectroscopy of the inactive filly oxidized form of the enzyme. The changes in the macrocycle vibrational modes are consistent with a Ca{sup 2+}-dependent increase in the out-of-plane distortion of the low-potential heme, the proposed peroxidatic heme. Most of the increase in out-of-plane distortion occurs when the high affinity site I is occupied, but a small further increase in distortion occurs when site II is also occupied by Ca{sup 2+}or Mg{sup 2+}. This increase in the heme distortion also explains the red shift in the Soret absorption band that occurs upon Ca{sup 2+} binding. Changes also occur in the low frequency substituent modes of the heme, indicating that a structural change in the covalently attached fingerprint pentapeptide of the LP heme occurs upon CM{sup 2+} binding to site I. These structural changes, possibly enhanced in the semi-reduced form of the enzyme, may lead to loss of the sixth ligand at the peroxidatic heme and activation of the enzyme.

  16. [Surgical management of limb prosthetic vascular graft exposure].

    Science.gov (United States)

    Wu, Hao; Wu, Wei-wei; Bai, Ming; Zeng, Rong; Song, Xiao-jun; Chen, Yu; Liu, Chang-wei

    2012-12-01

    To investigate the risk factors and surgical management of limb prosthetic vascular graft exposure. The clinical data of 17 patients suffering from limb prosthetic vascular graft exposure in Peking Union Medical College Hospital from August 2006 to December 2011 were retrospectively analyzed. Among 17 cases, 5 suffered from prosthetic vascular graft exposure after vascular bypass for the lower limb ischemia, whereas 12 were affected after the artificial graft arteriovenous fistula construction in the upper limbs for hemodialysis. The surgical procedures mainly included the local debridement as well as the local flap and transferred muscle-cutaneous flap reconstruction to preserve the prosthetic vascular graft. All 17 patients underwent local flap or muscle-cutaneous flap coverage procedure. After the surgery, the prosthetic vascular graft was successfully salvaged in 14 cases. The total successful rate was 82.4%. The surgery failed in three patients, in whom the prosthetic vascular grafts were finally removed. Local flap and transferred muscle-cutaneous flap reconstruction is an effective surgical management to salvage the exposed grafts.

  17. Instantaneous stiffness and hysteresis of dynamic elastic response prosthetic feet.

    Science.gov (United States)

    Webber, Christina M; Kaufman, Kenton

    2017-10-01

    Dynamic elastic response prosthetic feet are designed to mimic the functional characteristics of the native foot/ankle joint. Numerous designs of dynamic elastic response feet exist which make the prescription process difficult, especially because of the lack of empirical evidence describing the objective performance characteristics of the feet. To quantify the mechanical properties of available dynamic elastic response prosthetic feet, specifically the stiffness and hysteresis. Mechanical testing of dynamic elastic response prosthetic feet. Static Proof Testing in accordance with ISO 10328 was conducted on seven dynamic elastic response prosthetic feet. Load-displacement data were used to calculate the instantaneous stiffness in both the heel and forefoot regions, as well as hysteresis associated with each foot. Heel stiffness was greater than forefoot stiffness for all feet. The heel of the glass composite prosthetic foot was stiffer than the carbon fiber feet and it exhibited less hysteresis. Two different carbon fiber feet had the stiffest forefoot regions. Mechanical testing is a reproducible method that can be used to provide objective evidence about dynamic elastic response prosthetic foot performance and aid in the prescription process. Clinical relevance The quantitative stiffness and hysteresis data from this study can be used by prosthetists to aid the prescription process and make it more objective.

  18. Rotationplasty with vascular reconstruction for prosthetic knee joint infection.

    Science.gov (United States)

    Fujiki, Masahide; Miyamoto, Shimpei; Nakatani, Fumihiko; Kawai, Akira; Sakuraba, Minoru

    2015-01-01

    Rotationplasty is used most often as a function-preserving salvage procedure after resection of sarcomas of the lower extremity; however, it is also used after infection of prosthetic knee joints. Conventional vascular management during rotationplasty is to preserve and coil major vessels, but recently, transection and reanastomosis of the major vessels has been widely performed. However, there has been little discussion regarding the optimal vascular management of rotationplasty after infection of prosthetic knee joints because rotationplasty is rarely performed for this indication. We reviewed four patients who had undergone resection of osteosarcomas of the femur, placement of a prosthetic knee joint, and rotationplasty with vascular reconstruction from 2010 to 2013. The mean interval between prosthetic joint replacement and rotationplasty was 10.4 years and the mean interval between the diagnosis of prosthesis infection and rotationplasty was 7.9 years. Rotationplasty was successful in all patients; however, in one patient, arterial thrombosis developed and necessitated urgent surgical removal and arterial reconstruction. All patients were able to walk independently with a prosthetic limb after rehabilitation. Although there is no consensus regarding the most appropriate method of vascular management during rotationplasty for revision of infected prosthetic joints, vascular transection and reanastomosis is a useful option.

  19. Rotationplasty with Vascular Reconstruction for Prosthetic Knee Joint Infection

    Directory of Open Access Journals (Sweden)

    Masahide Fujiki

    2015-01-01

    Full Text Available Rotationplasty is used most often as a function-preserving salvage procedure after resection of sarcomas of the lower extremity; however, it is also used after infection of prosthetic knee joints. Conventional vascular management during rotationplasty is to preserve and coil major vessels, but recently, transection and reanastomosis of the major vessels has been widely performed. However, there has been little discussion regarding the optimal vascular management of rotationplasty after infection of prosthetic knee joints because rotationplasty is rarely performed for this indication. We reviewed four patients who had undergone resection of osteosarcomas of the femur, placement of a prosthetic knee joint, and rotationplasty with vascular reconstruction from 2010 to 2013. The mean interval between prosthetic joint replacement and rotationplasty was 10.4 years and the mean interval between the diagnosis of prosthesis infection and rotationplasty was 7.9 years. Rotationplasty was successful in all patients; however, in one patient, arterial thrombosis developed and necessitated urgent surgical removal and arterial reconstruction. All patients were able to walk independently with a prosthetic limb after rehabilitation. Although there is no consensus regarding the most appropriate method of vascular management during rotationplasty for revision of infected prosthetic joints, vascular transection and reanastomosis is a useful option.

  20. Characterization of the outer membrane receptor ShuA from the heme uptake system of Shigella dysenteriae. Substrate specificity and identification of the heme protein ligands.

    Science.gov (United States)

    Burkhard, Kimberly A; Wilks, Angela

    2007-05-18

    Shigella dysenteriae, like many bacterial pathogens, has evolved outer membrane receptor-mediated pathways for the uptake and utilization of heme as an iron source. As a first step toward understanding the mechanism of heme uptake we have undertaken a site-directed mutagenesis, spectroscopic, and kinetic analysis of the outer membrane receptor ShuA of S. dysenteriae. Purification of the outer membrane receptor gave a single band of molecular mass 73 kDa on SDS-PAGE. Initial spectroscopic analysis of the protein in either detergent micelles or lipid bicelles revealed residual heme bound to the receptor, with a Soret maximum at 413 nm. Titration of the protein with exogenous heme gave a Soret peak at 437 nm in detergent micelles, and 402 nm in lipid bicelles. However, transfer of heme from hemoglobin yields a Soret maximum at 413 nm identical to that of the isolated protein. Further spectroscopic and kinetic analysis revealed that hemoglobin in the oxidized state is the most likely physiological substrate for ShuA. In addition, mutation of the conserved histidines, H86A or H420A, resulted in a loss of the ability of the receptor to efficiently extract heme from hemoglobin. In contrast the double mutant H86A/H420A was unable to extract heme from hemoglobin. These findings taken together confirm that both His-86 and His-420 are essential for substrate recognition, heme coordination, and transfer. Furthermore, the full-length TonB was shown to form a 1:1 complex with either apo-ShuA H86A/H420A or the wild-type ShuA. These observations provide a basis for future studies on the coordination and transport of heme by the TonB-dependent outer membrane receptors.

  1. The five near-iron transporter (NEAT) domain anthrax hemophore, IsdX2, scavenges heme from hemoglobin and transfers heme to the surface protein IsdC.

    Science.gov (United States)

    Honsa, Erin Sarah; Fabian, Marian; Cardenas, Ana Maria; Olson, John S; Maresso, Anthony William

    2011-09-23

    Pathogenic bacteria require iron to replicate inside mammalian hosts. Recent studies indicate that heme acquisition in Gram-positive bacteria is mediated by proteins containing one or more near-iron transporter (NEAT) domains. Bacillus anthracis is a spore-forming, Gram-positive pathogen and the causative agent of anthrax disease. The rapid, extensive, and efficient replication of B. anthracis in host tissues makes this pathogen an excellent model organism for the study of bacterial heme acquisition. B. anthracis secretes two NEAT hemophores, IsdX1 and IsdX2. IsdX1 contains a single NEAT domain, whereas IsdX2 has five, a novel property among hemophores. To understand the functional significance of harboring multiple, non-identical NEAT domains, we purified each individual NEAT domain of IsdX2 as a GST fusion and analyzed the specific function of each domain as it relates to heme acquisition and transport. NEAT domains 1, 3, 4, and 5 all bind heme, with domain 5 having the highest affinity. All NEATs associate with hemoglobin, but only NEAT1 and -5 can extract heme from hemoglobin, seemingly by a specific and active process. NEAT1, -3, and -4 transfer heme to IsdC, a cell wall-anchored anthrax NEAT protein. These results indicate that IsdX2 has all the features required to acquire heme from the host and transport heme to the bacterial cell wall. Additionally, these results suggest that IsdX2 may accelerate iron import rates by acting as a "heme sponge" that enhances B. anthracis replication in iron-starved environments.

  2. The Five Near-iron Transporter (NEAT) Domain Anthrax Hemophore, IsdX2, Scavenges Heme from Hemoglobin and Transfers Heme to the Surface Protein IsdC*

    Science.gov (United States)

    Honsa, Erin Sarah; Fabian, Marian; Cardenas, Ana Maria; Olson, John S.; Maresso, Anthony William

    2011-01-01

    Pathogenic bacteria require iron to replicate inside mammalian hosts. Recent studies indicate that heme acquisition in Gram-positive bacteria is mediated by proteins containing one or more near-iron transporter (NEAT) domains. Bacillus anthracis is a spore-forming, Gram-positive pathogen and the causative agent of anthrax disease. The rapid, extensive, and efficient replication of B. anthracis in host tissues makes this pathogen an excellent model organism for the study of bacterial heme acquisition. B. anthracis secretes two NEAT hemophores, IsdX1 and IsdX2. IsdX1 contains a single NEAT domain, whereas IsdX2 has five, a novel property among hemophores. To understand the functional significance of harboring multiple, non-identical NEAT domains, we purified each individual NEAT domain of IsdX2 as a GST fusion and analyzed the specific function of each domain as it relates to heme acquisition and transport. NEAT domains 1, 3, 4, and 5 all bind heme, with domain 5 having the highest affinity. All NEATs associate with hemoglobin, but only NEAT1 and -5 can extract heme from hemoglobin, seemingly by a specific and active process. NEAT1, -3, and -4 transfer heme to IsdC, a cell wall-anchored anthrax NEAT protein. These results indicate that IsdX2 has all the features required to acquire heme from the host and transport heme to the bacterial cell wall. Additionally, these results suggest that IsdX2 may accelerate iron import rates by acting as a “heme sponge” that enhances B. anthracis replication in iron-starved environments. PMID:21808055

  3. Prosthetic status and treatment needs for lost masticatory function in haemodialysis patients.

    Science.gov (United States)

    Wilczyńska-Borawska, Magdalena; Małyszko, Jolanta; Cylwik-Rokicka, Dorota; Myśliwiec, Michał

    2012-02-29

    Premature loss of permanent teeth leads to stomatognathic system disability. It is a very serious but underrated problem for patients with chronic renal failure. The aim of study was analyse the degree of loss of masticatory function and number of teeth present for haemodialysis patients, and to define patients' needs for prosthetic treatment, which could restore correct occlusal condition. Sixty-nine haemodialysis patients treated at the Nephrology and Transplantology Clinic with the Dialysis Centre at the Medical University of Bialystok, Poland. We checked: 1) the total number of teeth and number of teeth separately for upper and lower jaws, 2) the existing prosthetic restorations and 3) the preserved masticatory function. More male than female patients were in possession of full dentition.All patients with at least 28 natural teeth with retained occlusal contacts whilst chewing were males (4; 10% males; 5.7% of the whole group). There were 15 edentulous patients: 7 males (10%) and 8 females (11.5%). Hundered percent of female patients presented with various degrees of tooth loss and needed prosthetic treatment. Nearly 70% of tested haemodialysis patients did not have a reconstructed masticatory function. The population of haemodialysis patients from the North East part of Poland are patients with severe stomatognathic system dysfunctions. It is of importance for dentists, as well as nephrologists, to understand the essence of the problem, as the general health of a patient cannot be improved without ensuring functional comfort of such as important system as the masticatory one.

  4. Clinical relevance of porphyrin determinations in porphyria

    NARCIS (Netherlands)

    Beukeveld, Gerhardus Johannes Jozef

    1995-01-01

    This thesis deals with hereditaryd iseasesin the synthesiso f heme. Heme presents a prosthetic group in a number of important proteins, like hemoglobin, cytochromes and the cytochrome P450 oxidases. In chapter 1 the heme synthesis and its regulation are described. Intermediary products in the

  5. O{sub 2}-mediated oxidation of ferrous nitrosylated human serum heme-albumin is limited by nitrogen monoxide dissociation

    Energy Technology Data Exchange (ETDEWEB)

    Ascenzi, Paolo, E-mail: ascenzi@uniroma3.it [Interdepartmental Laboratory of Electron Microscopy, University Roma Tre, Via della Vasca Navale 79, I-00146 Roma (Italy); National Institute for Infectious Diseases I.R.C.C.S. ' Lazzaro Spallanzani' , Via Portuense 292, I-00149 Roma (Italy); Gullotta, Francesca; Gioia, Magda; Coletta, Massimo [Department of Experimental Medicine and Biochemical Sciences, University of Roma ' Tor Vergata' , Via Montpellier 1, I-00133 Roma (Italy); Interuniversity Consortium for the Research on the Chemistry of Metals in Biological Systems, Piazza Umberto I 1, I-87100 Bari (Italy); Fasano, Mauro [Department of Structural and Functional Biology, and Center of Neuroscience, University of Insubria, Via Alberto da Giussano 12a, I-21052 Busto Arsizio, VA (Italy)

    2011-03-04

    Research highlights: {yields} Human serum heme-albumin displays globin-like properties. {yields} O{sub 2}-mediated oxidation of ferrous nitrosylated human serum heme-albumin. {yields} Allosteric modulation of human serum heme-albumin reactivity. {yields} Rifampicin is an allosteric effector of human serum heme-albumin. {yields} Human serum heme-albumin is a ROS and NOS scavenger. -- Abstract: Human serum heme-albumin (HSA-heme-Fe) displays globin-like properties. Here, kinetics of O{sub 2}-mediated oxidation of ferrous nitrosylated HSA-heme-Fe (HSA-heme-Fe(II)-NO) is reported. Values of the first-order rate constants for O{sub 2}-mediated oxidation of HSA-heme-Fe(II)-NO (i.e., for ferric HSA-heme-Fe formation) and for NO dissociation from HSA-heme-Fe(II)-NO (i.e., for NO replacement by CO) are k = 9.8 x 10{sup -5} and 8.3 x 10{sup -4} s{sup -1}, and h = 1.3 x 10{sup -4} and 8.5 x 10{sup -4} s{sup -1}, in the absence and presence of rifampicin, respectively, at pH = 7.0 and T = 20.0 {sup o}C. The coincidence of values of k and h indicates that NO dissociation represents the rate limiting step of O{sub 2}-mediated oxidation of HSA-heme-Fe(II)-NO. Mixing HSA-heme-Fe(II)-NO with O{sub 2} does not lead to the formation of the transient adduct(s), but leads to the final ferric HSA-heme-Fe derivative. These results reflect the fast O{sub 2}-mediated oxidation of ferrous HSA-heme-Fe and highlight the role of drugs in modulating allosterically the heme-Fe-atom reactivity.

  6. Dietary hemoglobin rescues young piglets from severe iron deficiency anemia: Duodenal expression profile of genes involved in heme iron absorption.

    Directory of Open Access Journals (Sweden)

    Robert Staroń

    Full Text Available Heme is an efficient source of iron in the diet, and heme preparations are used to prevent and cure iron deficiency anemia in humans and animals. However, the molecular mechanisms responsible for heme absorption remain only partially characterized. Here, we employed young iron-deficient piglets as a convenient animal model to determine the efficacy of oral heme iron supplementation and investigate the pathways of heme iron absorption. The use of bovine hemoglobin as a dietary source of heme iron was found to efficiently counteract the development of iron deficiency anemia in piglets, although it did not fully rebalance their iron status. Our results revealed a concerted increase in the expression of genes responsible for apical and basolateral heme transport in the duodenum of piglets fed a heme-enriched diet. In these animals the catalytic activity of heme oxygenase 1 contributed to the release of elemental iron from the protoporphyrin ring of heme within enterocytes, which may then be transported by the strongly expressed ferroportin across the basolateral membrane to the circulation. We hypothesize that the well-recognized high bioavailability of heme iron may depend on a split pathway mediating the transport of heme-derived elemental iron and intact heme from the interior of duodenal enterocytes to the bloodstream.

  7. A Prosthetic Foot Emulator to Optimize Prescription of Prosthetic Feet in Veterans and Service Members with Leg Amputations

    Science.gov (United States)

    2017-09-01

    on Microprocessor -Controlled Prosthetic Knees . Journal of Prosthetics and Orthotics. 2013;25(4S):53-55. 28. Raschke SU, Orendurff MS, Mattie JL, et al... Knee -Ankle Prosthesis Leg on Everyday Community Mobility and Social Interaction Role Site Principal Investigator (A Jayaraman, PI) (5% effort...Funding $2.5 M Purpose To evaluate new prosthesis that has powered controls at both the knee and ankle joints and a new way of controlling this device

  8. Surface EMG in advanced hand prosthetics.

    Science.gov (United States)

    Castellini, Claudio; van der Smagt, Patrick

    2009-01-01

    One of the major problems when dealing with highly dexterous, active hand prostheses is their control by the patient wearing them. With the advances in mechatronics, building prosthetic hands with multiple active degrees of freedom is realisable, but actively controlling the position and especially the exerted force of each finger cannot yet be done naturally. This paper deals with advanced robotic hand control via surface electromyography. Building upon recent results, we show that machine learning, together with a simple downsampling algorithm, can be effectively used to control on-line, in real time, finger position as well as finger force of a highly dexterous robotic hand. The system determines the type of grasp a human subject is willing to use, and the required amount of force involved, with a high degree of accuracy. This represents a remarkable improvement with respect to the state-of-the-art of feed-forward control of dexterous mechanical hands, and opens up a scenario in which amputees will be able to control hand prostheses in a much finer way than it has so far been possible.

  9. Identification of dynamic load for prosthetic structures.

    Science.gov (United States)

    Zhang, Dequan; Han, Xu; Zhang, Zhongpu; Liu, Jie; Jiang, Chao; Yoda, Nobuhiro; Meng, Xianghua; Li, Qing

    2017-12-01

    Dynamic load exists in numerous biomechanical systems, and its identification signifies a critical issue for characterizing dynamic behaviors and studying biomechanical consequence of the systems. This study aims to identify dynamic load in the dental prosthetic structures, namely, 3-unit implant-supported fixed partial denture (I-FPD) and teeth-supported fixed partial denture. The 3-dimensional finite element models were constructed through specific patient's computerized tomography images. A forward algorithm and regularization technique were developed for identifying dynamic load. To verify the effectiveness of the identification method proposed, the I-FPD and teeth-supported fixed partial denture structures were investigated to determine the dynamic loads. For validating the results of inverse identification, an experimental force-measuring system was developed by using a 3-dimensional piezoelectric transducer to measure the dynamic load in the I-FPD structure in vivo. The computationally identified loads were presented with different noise levels to determine their influence on the identification accuracy. The errors between the measured load and identified counterpart were calculated for evaluating the practical applicability of the proposed procedure in biomechanical engineering. This study is expected to serve as a demonstrative role in identifying dynamic loading in biomedical systems, where a direct in vivo measurement may be rather demanding in some areas of interest clinically. Copyright © 2017 John Wiley & Sons, Ltd.

  10. Prosthetic rehabilitation of the upper limb amputee

    Directory of Open Access Journals (Sweden)

    Bernard O′Keeffe

    2011-01-01

    Full Text Available The loss of all or part of the arm is a catastrophic event for a patient and a significant challenge to rehabilitation professionals and prosthetic engineers. The large, upper extremity amputee population in India has, historically, been poorly served, with most having no access to support or being provided with ineffective prostheses. In recent years, the arrival of organisations like Otto Bock has made high quality service standards and devices accessible to more amputees. This review attempts to provide surgeons and other medical professionals with an overview of the multidisciplinary, multistage rehabilitation process and the solution options available. With worldwide upper extremity prosthesis rejection rates at significant levels, the review also describes some of the factors which influence the outcome. This is particularly relevant in the Indian context where the service can involve high cost investments. It is the responsibility of all contributing professionals to guide vulnerable patients through the process and try to maximise the benefit that can be obtained within the resources available.

  11. Peri-prosthetic fracture vibration testing

    Energy Technology Data Exchange (ETDEWEB)

    Cruce, Jesse R [Los Alamos National Laboratory; Erwin, Jenny R [Los Alamos National Laboratory; Remick, Kevin R [Los Alamos National Laboratory; Cornwell, Phillip J [Los Alamos National Laboratory; Menegini, R. Michael [INDIANA UNIV.; Racanelli, Joe [STRYKER ORTHOPARDICS

    2010-11-08

    The purpose of this study was to establish a test setup and vibration analysis method to predict femoral stem seating and prevent bone fracture using accelerometer and force response data from an instrumented stem and impactor. This study builds upon earlier studies to identify a means to supplement a surgeon's tactile and auditory senses by using damage identification techniques normally used for civil and mechanical structures. Testing was conducted using foam cortical shell sawbones prepared for stems of different geometries. Each stem was instrumented with an accelerometer. Two impactor designs were compared: a monolithic impactor and a two-piece impactor, each with an integrated load cell and accelerometer. Acceleration and force measurements were taken in the direction of impaction. Comparisons between different methods of applying an impacting force were made, including a drop tower and a surgical hammer. The effect of varying compliance on the data was also investigated. The ultimate goal of this study was to assist in the design of an integrated portable data acquisition system capable of being used in future cadaveric testing. This paper will discuss the experimental setup and the subsequent results of the comparisons made between impactors, prosthetic geometries, compliances, and impact methods. The results of this study can be used for both future replicate testing as well as in a cadaveric environment.

  12. Monitoring oral iron therapy with protoporphyrin/heme ratios in pregnant women.

    Science.gov (United States)

    Madan, N; Prasannaraj, P; Rusia, U; Sundaram, K R; Nath, L M; Sood, S K

    1999-06-01

    Assessment of the efficacy of iron therapy has usually been done in populations/patients by monitoring changes in hemoglobin concentration, serum iron, percent transferrin saturation, and serum ferritin. In this study the protoporphyrin heme (P/H) ratio (a measure of free erythrocyte protoporphyrin) was measured before and after iron therapy in three groups of pregnant women, who received 60 mg (group A), 120 mg (group B), and 240 mg (group C) of elemental iron with folic acid (0.5 mg) per day for a period of 12 weeks, to evaluate its efficacy to monitor iron therapy. The three groups were comparable regarding the initial mean Hb concentration and serum ferritin levels. The initial mean P/H ratios were markedly elevated in all three groups and were different in the three groups, being highest in group A (113.2+/-92.6), intermediate in group B (87.5+/-62.5), and lowest in group C (69.8+/-43.3). The initial P/H ratio was significantly higher in group A than in group C (p<0.05). This probably affected the efficacy of iron therapy in the three groups. The P/H ratio decreased significantly in each of the three groups after iron therapy (A and B: p<0.001; C p<0.01). Mean Hb concentration and serum ferritin increased in all three groups post therapy; however, the magnitude of change in P/H ratio in all three groups was much greater. This indicated that the predominant contributory factor for anemia was iron deficiency in this group of pregnant women. Serum iron and percent transferrin saturation are difficult to interpret in our population, as iron is freely available over the counter and is prescribed as soon as anemia is detected in patients; therefore, the reduction in P/H ratio may be used to monitor response to iron therapy in population groups.

  13. Chlamydomonas reinhardtii LFO1 Is an IsdG Family Heme Oxygenase.

    Science.gov (United States)

    Lojek, Lisa J; Farrand, Allison J; Wisecaver, Jennifer H; Blaby-Haas, Crysten E; Michel, Brian W; Merchant, Sabeeha S; Rokas, Antonis; Skaar, Eric P

    2017-01-01

    Heme is essential for respiration across all domains of life. However, heme accumulation can lead to toxicity if cells are unable to either degrade or export heme or its toxic by-products. Under aerobic conditions, heme degradation is performed by heme oxygenases, enzymes which utilize oxygen to cleave the tetrapyrrole ring of heme. The HO-1 family of heme oxygenases has been identified in both bacterial and eukaryotic cells, whereas the IsdG family has thus far been described only in bacteria. We identified a hypothetical protein in the eukaryotic green alga Chlamydomonas reinhardtii, which encodes a protein containing an antibiotic biosynthesis monooxygenase (ABM) domain consistent with those associated with IsdG family members. This protein, which we have named LFO1, degrades heme, contains similarities in predicted secondary structures to IsdG family members, and retains the functionally conserved catalytic residues found in all IsdG family heme oxygenases. These data establish LFO1 as an IsdG family member and extend our knowledge of the distribution of IsdG family members beyond bacteria. To gain further insight into the distribution of the IsdG family, we used the LFO1 sequence to identify 866 IsdG family members, including representatives from all domains of life. These results indicate that the distribution of IsdG family heme oxygenases is more expansive than previously appreciated, underscoring the broad relevance of this enzyme family. IMPORTANCE This work establishes a protein in the freshwater alga Chlamydomonas reinhardtii as an IsdG family heme oxygenase. This protein, LFO1, exhibits predicted secondary structure and catalytic residues conserved in IsdG family members, in addition to a chloroplast localization sequence. Additionally, the catabolite that results from the degradation of heme by LFO1 is distinct from that of other heme degradation products. Using LFO1 as a seed, we performed phylogenetic analysis, revealing that the IsdG family is

  14. Iron-coordinating tyrosine is a key determinant of NEAT domain heme transfer.

    Science.gov (United States)

    Grigg, Jason C; Mao, Cherry X; Murphy, Michael E P

    2011-10-28

    In humans, heme iron is the most abundant iron source, and bacterial pathogens such as Staphylococcus aureus acquire it for growth. IsdB of S. aureus acquires Fe(III)-protoporphyrin IX (heme) from hemoglobin for transfer to IsdC via IsdA. These three cell-wall-anchored Isd (iron-regulated surface determinant) proteins contain conserved NEAT (near iron transport) domains. The purpose of this work was to delineate the mechanism of heme binding and transfer between the NEAT domains of IsdA, IsdB, and IsdC using a combination of structural and spectroscopic studies. X-ray crystal structures of IsdA NEAT domain (IsdA-N1) variants reveal that removing the native heme-iron ligand Tyr166 is compensated for by iron coordination by His83 on the distal side and that no single mutation of distal loop residues is sufficient to perturb the IsdA-heme complex. Also, alternate heme-iron coordination was observed in structures of IsdA-N1 bound to reduced Fe(II)-protoporphyrin IX and Co(III)-protoporphyrin IX. The IsdA-N1 structural data were correlated with heme transfer kinetics from the NEAT domains of IsdB and IsdC. We demonstrated that the NEAT domains transfer heme at rates comparable to full-length proteins. The second-order rate constant for heme transfer from IsdA-N1 was modestly affected (15-fold (to 100-fold excess IsdC). We propose a heme transfer model wherein NEAT domain complexes pass heme iron directly from an iron-coordinating Tyr of the donor protein to the homologous Tyr residues of the acceptor protein. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. Biosocial profile of New Zealand prosthetic eye wearers.

    Science.gov (United States)

    Pine, Keith R; Sloan, Brian; Jacobs, Robert J

    2012-10-12

    To describe the biosocial profile of New Zealand (NZ) artificial eye wearers and establish a basis for future research and international comparison. This retrospective study surveyed 431 NZ artificial eye wearers to investigate their ethnicity, gender, age, causes of eye loss, age of current prosthesis, ocular prosthetic maintenance regimes and the extent and severity of discharge associated with prosthesis wear. Approximately 3000 people wear artificial eyes in NZ. Accidents were the main cause of eye loss prior to 1990 and medical conditions have been the main cause since. In the 1960s, the ratio of men to women losing an eye from accidents was 5:1, but during the past decade the ratio was 1.4:1. Socket discharge occurred at least twice daily for one-third of the study group. Approximately 1 in 1440 people wear artificial eyes in NZ. Decline of eye loss due to accidents is consistent with decreasing workplace and traffic accidents and may be due to improved medical management, workplace safety standards and safer roads. Mucoid discharge is prevalent in the anophthalmic population of NZ and an evidence based treatment protocol for discharge associated with prosthesis wear is needed. Research into this distressing condition is planned.

  16. Pilot-scale tests of HEME and HEPA dissolution process

    Energy Technology Data Exchange (ETDEWEB)

    Qureshi, Z.H.; Strege, D.K.

    1994-06-01

    A series of pilot-scale demonstration tests for the dissolution of High Efficiency Mist Eliminators (HEME`s) and High Efficiency Particulate Airfilters (HEPA) were performed on a 1/5th linear scale. These fiberglass filters are to be used in the Defense Waste Processing Facility (DWPF) to decontaminate the effluents from the off-gases generated during the feed preparation process and vitrification. When removed, these filters will be dissolved in the Decontamination Waste Treatment Tank (DWTT) using 5 wt% NaOH solution. The contaminated fiberglass is converted to an aqueous stream which will be transferred to the waste tanks. The filter metal structure will be rinsed with process water before its disposal as low-level solid waste. The pilot-scale study reported here successfully demonstrated a simple one step process using 5 wt% NaOH solution. The proposed process requires the installation of a new water spray ring with 30 nozzles. In addition to the reduced waste generated, the total process time is reduced to 48 hours only (66% saving in time). The pilot-scale tests clearly demonstrated that the dissolution process of HEMEs has two stages - chemical digestion of the filter and mechanical erosion of the digested filter. The digestion is achieved by a boiling 5 wt% caustic solutions, whereas the mechanical break down of the digested filter is successfully achieved by spraying process water on the digested filter. An alternate method of breaking down the digested filter by increased air sparging of the solution was found to be marginally successful are best. The pilot-scale tests also demonstrated that the products of dissolution are easily pumpable by a centrifugal pump.

  17. Heme oxygenase-1 accelerates tumor angiogenesis of human pancreatic cancer.

    Science.gov (United States)

    Sunamura, Makoto; Duda, Dan G; Ghattas, Maivel H; Lozonschi, Lucian; Motoi, Fuyuhiko; Yamauchi, Jun-Ichiro; Matsuno, Seiki; Shibahara, Shigeki; Abraham, Nader G

    2003-01-01

    Angiogenesis is necessary for the continued growth of solid tumors, invasion and metastasis. Several studies clearly showed that heme oxygenase-1 (HO-1) plays an important role in angiogenesis. In this study, we used the vital microscope system, transparent skinfold model, lung colonization model and transduced pancreatic cancer cell line (Panc-1)/human heme oxygenase-1 (hHO-1) cells, to precisely analyze, for the first time, the effect of hHO-1 gene on tumor growth, angiogenesis and metastasis. Our results revealed that HO-1 stimulates angiogenesis of pancreatic carcinoma in severe combined immune deficient mice. Overexpression of human hHO-1 after its retroviral transfer into Panc-1 cells did not interfere with tumor growth in vitro. While in vivo the development of tumors was accelerated upon transfection with hHO-1. On the other hand, inhibition of heme oxygenase (HO) activity by stannous mesoporphyrin was able transiently to delay tumor growth in a dose dependent manner. Tumor angiogenesis was markedly increased in Panc-1/hHO-1 compared to mock transfected and wild type. Lectin staining and Ki-67 proliferation index confirmed these results. In addition hHO-1 stimulated in vitro tumor angiogenesis and increased endothelial cell survival. In a lung colonization model, overexpression of hHO-1 increased the occurrence of metastasis, while inhibition of HO activity by stannous mesoporphyrin completely inhibited the occurrence of metastasis. In conclusion, overexpression of HO-1 genes potentiates pancreatic cancer aggressiveness, by increasing tumor growth, angiogenesis and metastasis and that the inhibition of the HO system may be of useful benefit for the future treatment of the disease.

  18. A new method for photodynamic disinfection of prosthetic constructions and impressions in prosthetic dentistry.

    Science.gov (United States)

    Vlahova, Angelina P; Kisov, Christo K; Popova, Elka V; Haydushka, Irina A; Mantareva, Vanya N

    2012-01-01

    Photodynamic therapy is a topical treatment of pathogens that involves the use of a photoactive dye (photosensitizer), which is non-toxic when not exposed to light and activated by light of a specific wavelength in the presence of oxygen. The highly cytotoxic oxygen species generated by the induced photophysical processes inactivate the pathogenic cells. The PURPOSE of this study was to present a new method we developed for photodynamic disinfection of prostheses and impressions in prosthetic dentistry and to assess its effectiveness in comparison with some conventional methods of disinfection. The method was developed on the basis of series of experimental studies (30 experiments for each type of disinfectant, 30 controls with no disinfection for each material, and 30 direct cultures of each test microorganism--MRSA, P. aeruginosa and C. albicans) using standard test specimens made of prosthesis plastic and impression materials. The new method of photodynamic disinfection with GaPc1 as photosensitizer was 100% efficient in C-silicones, A-silicones and polyethers, but not in alginates (40%). To plastics the photodynamic method shows the same efficiency as the conventional disinfectants of hypochlorite solutions and denture cleansing tablets (100% effect). The method of photodynamic disinfection we developed is a good therapeutic choice against orally transmitted diseases in prosthetic dentistry.

  19. Quantification of Shear Stresses Within a Transtibial Prosthetic Socket.

    Science.gov (United States)

    Schiff, Adam; Havey, Robery; Carandang, Gerard; Wickman, Amy; Angelico, John; Patwardhan, Avinash; Pinzur, Michael

    2014-08-01

    There is a paucity of objectively recorded data delineating the pattern of weightbearing distribution within the prosthetic socket of patients with transtibial amputation. Our current knowledge is based primarily on information obtained from finite element analysis computer models. Four high-functioning transtibial amputees were fit with similar custom prosthetic sockets. Three load cells were incorporated into each socket at high stress contact areas predicted by computer modeling. Dynamic recording of prosthetic socket loading was accomplished during rising from a sitting position, stepping from a 2-leg stance to a 1-leg stance, and during the initiation of walking. By comparing the loads measured at each of the 3 critical locations, anterior/posterior shear, superior/inferior shear, and end weightbearing were recorded. The same load pattern in all 4 subjects was found during each of the 3 functional activities. The load transmission at the distal end of the amputation residual limbs was negligible. Consistent forces were observed in both the anterior/posterior and superior/inferior planes. Correlation coefficients were used to compare the loads measured in each of the 4 subjects, which ranged from a low of .82 to a high of .98, where a value approaching 1.0 implies a linear relationship amongst subjects. This experimental model appears to have accurately recorded loading within a transtibial prosthetic socket consistent with previously reported finite element analysis computer models. This clinical model will allow objective measurement of weightbearing within the prosthetic socket of transtibial amputees and allow objective comparison of weightbearing distribution within the prosthetic sockets of patients who have undergone creation of different versions of a transtibial amputation residual limb and prosthetic socket designs. © The Author(s) 2014.

  20. Efficacy of proprioceptive neuromuscular facilitation techniques versus traditional prosthetic training for improving ambulatory function in transtibial amputees

    Directory of Open Access Journals (Sweden)

    Pallavi Sahay, MPT

    2014-06-01

    Full Text Available The objective of this randomized controlled trial was to evaluate the efficacy of proprioceptive neuromuscular facilitation (PNF techniques in comparison to traditional prosthetic training (TPT in improving ambulatory function in transtibial amputees. Thirty study participants (19 men and 11 women with unilateral transtibial amputation participated in the study. They were randomly allocated to either the traditional training group (i.e., TPT (n = 15 or the PNF training group (n = 15. The treatment in the TPT group consisted of weight-bearing, weight-shifting, balance, and gait exercises for 30 minutes daily for 10 treatment sessions. In the PNF group, the same activities were performed by employing PNF principles and techniques. The outcome measures were gait parameters (e.g., stride width, step length, and stride length and the Locomotor Capabilities Index (LCI. The between-group comparisons at the end of the trial showed that the PNF group showed significant improvement in gait parameters and in the LCI, compared to the TPT group (p < 0.05. The results of the study suggested that prosthetic training based on proprioceptive feedback is more effective than the traditional prosthetic programme in improving ambulatory function.

  1. Hydrogen peroxide-mediated conversion of coproheme to heme b by HemQ-lessons from the first crystal structure and kinetic studies.

    Science.gov (United States)

    Hofbauer, Stefan; Mlynek, Georg; Milazzo, Lisa; Pühringer, Dominic; Maresch, Daniel; Schaffner, Irene; Furtmüller, Paul G; Smulevich, Giulietta; Djinović-Carugo, Kristina; Obinger, Christian

    2016-12-01

    Heme biosynthesis in Gram-positive bacteria follows a recently described coproporphyrin-dependent pathway with HemQ catalyzing the decarboxylation of coproheme to heme b. Here we present the first crystal structure of a HemQ (homopentameric coproheme-HemQ from Listeria monocytogenes) at 1.69 Å resolution and the conversion of coproheme to heme b followed by UV-vis and resonance Raman spectroscopy as well as mass spectrometry. The ferric five-coordinated coproheme iron of HemQ is weakly bound by a neutral proximal histidine H174. In the crystal structure of the resting state, the distal Q187 (conserved in Firmicutes HemQ) is H-bonded with propionate p2 and the hydrophobic distal cavity lacks solvent water molecules. Two H 2 O 2 molecules are shown to be necessary for decarboxylation of the propionates p2 and p4, thereby forming the corresponding vinyl groups of heme b. The overall reaction is relatively slow (k cat /K M = 1.8 × 10 2 m -1 ·s -1 at pH 7.0) and occurs in a stepwise manner with a three-propionate intermediate. We present the noncovalent interactions between coproheme and the protein and propose a two-step reaction mechanism. Furthermore, the structure of coproheme-HemQ is compared to that of the phylogenetically related heme b-containing chlorite dismutases. Structural data are available in the PDB under the accession number 5LOQ. © 2016 The Authors The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

  2. Cloning and Expression of cDNA for Rat Heme Oxygenase

    Science.gov (United States)

    Shibahara, Shigeki; Muller, Rita; Taguchi, Hayao; Yoshida, Tadashi

    1985-12-01

    Two cDNA clones for rat heme oxygenase have been isolated from a rat spleen cDNA library in λ gt11 by immunological screening using a specific polyclonal antibody. One of these clones has an insert of 1530 nucleotides that contains the entire protein-coding region. To confirm that the isolated cDNA encodes heme oxygenase, we transfected monkey kidney cells (COS-7) with the cDNA carried in a simian virus 40 vector. The heme oxygenase was highly expressed in endoplasmic reticulum of transfected cells. The nucleotide sequence of the cloned cDNA was determined and the primary structure of heme oxygenase was deduced. Heme oxygenase is composed of 289 amino acids and has one hydrophobic segment at its carboxyl terminus, which is probably important for the insertion of heme oxygenase into endoplasmic reticulum. The cloned cDNA was used to analyze the induction of heme oxygenase in rat liver by treatment with CoCl2 or with hemin. RNA blot analysis showed that both CoCl2 and hemin increased the amount of hybridizable mRNA, suggesting that these substances may act at the transcriptional level to increase the amount of heme oxygenase.

  3. Stanniocalcin 1 binds hemin through a partially conserved heme regulatory motif

    Energy Technology Data Exchange (ETDEWEB)

    Westberg, Johan A., E-mail: johan.westberg@helsinki.fi [Department of Pathology, Haartman Institute, University of Helsinki and HUSLAB, P.O. Box 21, Haartmaninkatu 3, FI-00014 Helsinki (Finland); Jiang, Ji, E-mail: ji.jiang@helsinki.fi [Department of Pathology, Haartman Institute, University of Helsinki and HUSLAB, P.O. Box 21, Haartmaninkatu 3, FI-00014 Helsinki (Finland); Andersson, Leif C., E-mail: leif.andersson@helsinki.fi [Department of Pathology, Haartman Institute, University of Helsinki and HUSLAB, P.O. Box 21, Haartmaninkatu 3, FI-00014 Helsinki (Finland)

    2011-06-03

    Highlights: {yields} Stanniocalcin 1 (STC1) binds heme through novel heme binding motif. {yields} Central iron atom of heme and cysteine-114 of STC1 are essential for binding. {yields} STC1 binds Fe{sup 2+} and Fe{sup 3+} heme. {yields} STC1 peptide prevents oxidative decay of heme. -- Abstract: Hemin (iron protoporphyrin IX) is a necessary component of many proteins, functioning either as a cofactor or an intracellular messenger. Hemoproteins have diverse functions, such as transportation of gases, gas detection, chemical catalysis and electron transfer. Stanniocalcin 1 (STC1) is a protein involved in respiratory responses of the cell but whose mechanism of action is still undetermined. We examined the ability of STC1 to bind hemin in both its reduced and oxidized states and located Cys{sup 114} as the axial ligand of the central iron atom of hemin. The amino acid sequence differs from the established (Cys-Pro) heme regulatory motif (HRM) and therefore presents a novel heme binding motif (Cys-Ser). A STC1 peptide containing the heme binding sequence was able to inhibit both spontaneous and H{sub 2}O{sub 2} induced decay of hemin. Binding of hemin does not affect the mitochondrial localization of STC1.

  4. The effect of irradiation and thermal process on beef heme iron concentration and color properties

    Energy Technology Data Exchange (ETDEWEB)

    Mistura, Liliana Perazzini Furtado; Colli, Celia [Universidade de Sao Paulo (USP), SP (Brazil). Faculdade de Ciencias Farmaceuticas. Dept. de Alimentos e Nutricao Experimental], e-mail: cecolli@usp.br

    2009-01-15

    The aim of this study was to evaluate the influence of irradiation and thermal process on the heme iron (heme-Fe) concentration and color properties of Brazilian cattle beef. Beef samples (patties and steaks) were irradiated at 0-10 kGy and cooked in a combination oven at 250 deg C for 9 minutes with 70% humidity. Total iron and heme iron (heme-Fe) concentrations were determined. The data were compared by multiple comparisons and fixed- effects ANOVA. Irradiation at doses higher than 5 kGy significantly altered the heme-Fe concentration. However, the sample preparation conditions interfered more in the heme-Fe content than did the irradiation. Depending on the animal species, meat heme iron levels between 35 and 52% of the total iron are used for dietetic calculations. In this study the percentage of heme-iron was, on average, 70% of the total iron showing that humidity is an important factor for its preservation. The samples were analyzed instrumentally for CIE L{sup *}, a{sup *}, and b{sup *} values. (author)

  5. Dietary heme adversely affects experimental colitis in rats, despite heat-shock protein induction

    NARCIS (Netherlands)

    Schepens, Marloes A. A.; Vink, Carolien; Schonewille, Arjan J.; Dijkstra, Gerard; van der Meer, Roelof; Bovee-Oudenhoven, Ingeborg M. J.

    Objective: Research on dietary modulation of inflammatory bowel disease is in its infancy. Dietary heme, mimicking red meat, is cytotoxic to colonic epithelium and thus may aggravate colitis. Alternatively, heme-induced colonic stress might also result in potential protective heat-shock proteins

  6. Long-term dietary heme iron and red meat intake in relation to endometrial cancer risk

    NARCIS (Netherlands)

    Genkinger, J.M.; Friberg, E.; Goldbohm, R.A.; Wolk, A.

    2012-01-01

    Background: Heme and total iron, present in meat, have been hypothesized to promote carcinogenesis. Few prospective studies have examined the associations between intakes of heme and total iron, types of meat, and endometrial cancer risk. Objective: We evaluated the associations between intakes of

  7. Unsaturated Glycerophospholipids Mediate Heme Crystallization: Biological Implications for Hemozoin Formation in the Kissing Bug Rhodnius prolixus

    DEFF Research Database (Denmark)

    Stiebler, R.; Majerowicz, David; Knudsen, Jens

    2014-01-01

    Hemozoin (Hz) is a heme crystal produced by some blood-feeding organisms, as an efficient way to detoxify heme derived from hemoglobin digestion. In the triatomine insect Rhodnius prolixus, Hz is essentially produced by midgut extracellular phospholipid membranes known as perimicrovillar membranes...

  8. Dietary heme modulates microbiota and mucosa of mouse colon without significant host-microbe cross talk

    NARCIS (Netherlands)

    IJssennagger, Noortje; Rijnierse, A.; Muller, Michael; Meer, van der Roelof

    2013-01-01

    Previously, we showed that dietary heme injured the colonic surface epithelium and induced hyperproliferation by changing the surface to crypt signaling. In this study we investigated whether bacteria play a role in this changed signaling. Dietary heme increased the Bacteroidetes and decreased the

  9. Natural chlorophyll but not chlorophyllin prevents heme-induced cytotoxic and hyperproliferative effects in rat colon

    NARCIS (Netherlands)

    Vogel, de J.; Jonker-Termont, D.S.M.L.; Katan, M.B.; Meer, van der R.

    2005-01-01

    Diets high in red meat and low in green vegetables are associated with an increased risk of colon cancer. In rats, dietary heme, mimicking red meat, increases colonic cytotoxicity and proliferation of the colonocytes, whereas addition of chlorophyll from green vegetables inhibits these heme-induced

  10. Heme and chlorophyll intake and risk of colorectal cancer in the Netherlands cohort study

    NARCIS (Netherlands)

    Balder, H.F.; Vogel, J. de; Jansen, M.C.J.F.; Weijenberg, M.P.; Brandt, P.A. van den; Westenbrink, S.; Meer, R.D. van der; Goldbohm, R.A.

    2006-01-01

    Background: The evidence for red meat as a determinant of colorectal cancer remains equivocal, which might be explained by differences in heme content. Heme is the prooxidant, iron-containing porphyrin pigment of meat and its content depends on the type of meat. Chlorophyll from green vegetables

  11. Kinematic and kinetic comparisons of transfemoral amputee gait using C-Leg and Mauch SNS prosthetic knees.

    Science.gov (United States)

    Segal, Ava D; Orendurff, Michael S; Klute, Glenn K; McDowell, Martin L; Pecoraro, Janice A; Shofer, Jane; Czerniecki, Joseph M

    2006-01-01

    The C-Leg (Otto Bock, Duderstadt, Germany) is a microprocessor-controlled prosthetic knee that may enhance amputee gait. This intrasubject randomized study compared the gait biomechanics of transfemoral amputees wearing the C-Leg with those wearing a common noncomputerized prosthesis, the Mauch SNS (Ossur, Reykjavik, Iceland). After subjects had a 3-month acclimation period with each prosthetic knee, typical gait biomechanical data were collected in a gait laboratory. At a controlled walking speed (CWS), peak swing phase knee-flexion angle decreased for the C-Leg group compared with the Mauch SNS group (55.2 degrees +/- 6.5 degrees vs 64.41 degrees +/- 5.8 degrees , respectively; p = 0.005); the C-Leg group was similar to control subjects' peak swing knee-flexion angle (56.0 degrees +/- 3.4 degrees ). Stance knee-flexion moment increased for the C-Leg group compared with the Mauch SNS group (0.142 +/- 0.05 vs 0.067 +/- 0.07 N"m, respectively; p = 0.01), but remained significantly reduced compared with control subjects (0.477 +/- 0.1 N"m). Prosthetic limb step length at CWS was less for the C-Leg group compared with the Mauch SNS group (0.66 +/- 0.04 vs 0.70 +/- 0.06 m, respectively; p = 0.005), which resulted in increased symmetry between limbs for the C-Leg group. Subjects also walked faster with the C-Leg versus the Mauch SNS (1.30 +/- 0.1 vs 1.21 +/- 0.1 m/s, respectively; p = 0.004). The C-Leg prosthetic limb vertical ground reaction force decreased compared with the Mauch SNS (96.3 +/- 4.7 vs 100.3 +/- 7.5 % body weight, respectively; p = 0.0092).

  12. Cigarette smoking and dental implant tooth replacement therapy: A questionnaire survey among patients receiving implant prosthetic treatment

    Directory of Open Access Journals (Sweden)

    Kubic-Filiks Beata

    2014-12-01

    Full Text Available Chronic nicotinism has negative effects, both local and systemic. Its local effects are related to both the immediate thermal influence, as well as the toxic action of the substances contained in the smoke. In addition, the microflora colonizing dental plague is changed. The damage and the inflammatory processes that are incurred, affect the bone tissue of the alveolar processes, the mucosa, gums, and the tooth enamel. In this study, the tobacco smoking-related profile of patients being treated by way of implants was determined. Moreover, the relationship between cigarette smoking and pain sensation was assessed in patients undergoing surgical and prosthetic procedures in the oral cavity. The questionnaire survey covered 464 patients receiving prosthetic treatment at the “Dental” Non-Public Health Care Centre in Tomaszow Mazowiecki. The patients answered questions concerning their sex, age, the period of smoking, number of cigarettes smoked per day and the sensation of pain during bone reconstruction, implant placement and prosthetic procedures. The most numerous group of patients treated with implants were women: either non-smoking or smoking for less than 20 years at a level of less than 20 cigarettes a day, and men aged 40-60 years who have been smoking for over 20 years, at more than 20 cigarettes a day. The results of the survey reveal that non-smoking patients felt pain during bone reconstruction, implant placement and prosthetic procedures more frequently.

  13. Recent advancements in prosthetic hand technology.

    Science.gov (United States)

    Saikia, Angana; Mazumdar, Sushmi; Sahai, Nitin; Paul, Sudip; Bhatia, Dinesh; Verma, Suresh; Rohilla, Punit Kumar

    2016-07-01

    Recently, significant advances over the past decade have been made in robotics, artificial intelligence and other cognitive related fields, allowing development of highly sophisticated bio-mimetic robotics systems. In addition, enormous number of robots have been designed and assembled by explicitly realising their biological oriented behaviours. To enhance skill behaviours and adequate grasping abilities in these devices, a new phase of dexterous hands has been developed recently with bio-mimetically oriented and bio-inspired functionalities. The aim in writing this review paper is to present a detailed insight towards the development of the bio-mimetic based dexterous robotic multi-fingered artificial hand. An "ideal" upper limb prosthesis should be perceived as a part of their natural body by the amputee and should replicate sensory-motor capabilities of the amputated limb. Upper-limb amputations are most often the result of sudden trauma to the body, although they also can be caused by malignancy, congenital deficiencies and vascular diseases. This paper discusses the different bio-mimetic approaches using a framework that permits for a common description of biological and technical based hand manipulation behaviour. In particular, the review focuses on a number of developments in the inspired robotic systems. In conclusion, the study found that a huge amount of research efforts in terms of kinematics, dynamics, modelling and control methodologies are being put in to improve the present hand technology, thereby providing more functionality to the prosthetic limb of the amputee. This would improve their quality-of-life and help in performing activities of daily living (ADL) tasks with comparative ease in the near future.

  14. Cyanide does more to inhibit heme enzymes, than merely serving as an active-site ligand

    Energy Technology Data Exchange (ETDEWEB)

    Parashar, Abhinav [Center for Biomedical Research, VIT University, Vellore, Tamil Nadu, 632014 India (India); Venkatachalam, Avanthika [REDOx Lab, PSG Institute of Advanced Studies, Avinashi Road, Peelamedu, Coimbatore, Tamil Nadu, 641004 (India); Gideon, Daniel Andrew [Center for Biomedical Research, VIT University, Vellore, Tamil Nadu, 632014 India (India); Manoj, Kelath Murali, E-mail: satyamjayatu@yahoo.com [REDOx Lab, PSG Institute of Advanced Studies, Avinashi Road, Peelamedu, Coimbatore, Tamil Nadu, 641004 (India)

    2014-12-12

    Highlights: • Cyanide (CN) is a well-studied toxic principle, known to inhibit heme-enzymes. • Inhibition is supposed to result from CN binding at the active site as a ligand. • Diverse heme enzymes’ CN inhibition profiles challenge prevailing mechanism. • Poor binding efficiency of CN at low enzyme concentrations and ligand pressures. • CN-based diffusible radicals cause ‘non-productive electron transfers’ (inhibition). - Abstract: The toxicity of cyanide is hitherto attributed to its ability to bind to heme proteins’ active site and thereby inhibit their activity. It is shown herein that the long-held interpretation is inadequate to explain several observations in heme-enzyme reaction systems. Generation of cyanide-based diffusible radicals in heme-enzyme reaction milieu could shunt electron transfers (by non-active site processes), and thus be detrimental to the efficiency of oxidative outcomes.

  15. Regiospecificity determinants of human heme oxygenase: differential NADPH- and ascorbate-dependent heme cleavage by the R183E mutant.

    Science.gov (United States)

    Wang, Jinling; Lad, Latesh; Poulos, Thomas L; Ortiz de Montellano, Paul R

    2005-01-28

    The ability of the human heme oxygenase-1 (hHO-1) R183E mutant to oxidize heme in reactions supported by either NADPH-cytochrome P450 reductase or ascorbic acid has been compared. The NADPH-dependent reaction, like that of wild-type hHO-1, yields exclusively biliverdin IXalpha. In contrast, the R183E mutant with ascorbic acid as the reductant produces biliverdin IXalpha (79 +/- 4%), IXdelta (19 +/- 3%), and a trace of IXbeta. In the presence of superoxide dismutase and catalase, the yield of biliverdin IXdelta is decreased to 8 +/- 1% with a corresponding increase in biliverdin IXalpha. Spectroscopic analysis of the NADPH-dependent reaction shows that the R183E ferric biliverdin complex accumulates, because reduction of the iron, which is required for sequential iron and biliverdin release, is impaired. Reversal of the charge at position 183 makes reduction of the iron more difficult. The crystal structure of the R183E mutant, determined in the ferric and ferrous-NO bound forms, shows that the heme primarily adopts the same orientation as in wild-type hHO-1. The structure of the Fe(II).NO complex suggests that an altered active site hydrogen bonding network supports catalysis in the R183E mutant. Furthermore, Arg-183 contributes to the regiospecificity of the wild-type enzyme, but its contribution is not critical. The results indicate that the ascorbate-dependent reaction is subject to a lower degree of regiochemical control than the NADPH-dependent reaction. Ascorbate may be able to reduce the R183E ferric and ferrous dioxygen complexes in active site conformations that cannot be reduced by NADPH-cytochrome P450 reductase.

  16. [Genistein attenuates monocrotaline-induced pulmonary arterial hypertension in rats by up-regulating heme oxygenase-1 expression].

    Science.gov (United States)

    Zhang, Yukun; Wang, Daoxin; Zhu, Tao; Li, Changyi

    2012-02-01

    To study the effect of genistein on the expression of heme oxygenase-1 (HO-1) in rats with pulmonary arterial hypertension (PAH) induced by monocrotaline (MCT). Sixty male Sprague-Dawley rats were randomly divided into 4 groups (n=15), namely the control group, model group, low-dose (20 µg/kg) genistein group and high-dose (80 µg/kg) genistein group. The hemodynamic parameters were measured and the remodeling of pulmonary small arteries was observed by electron microscope (EM). The expression of HO-1 in the lung tissues were detected by Western blotting. Compared with the model group, genistein treatment significantly reduced the elevated mean pulmonary arterial pressure, improved the right ventricular hypertrophy index, and increased the expression of HO-1 in a dose-dependent manner. Genistein attentuates pulmonary arterial hypertension in MCT-treated rats possibly by up-regulation of HO-1 in the lung tissues.

  17. Methods for characterization of mechanical and electrical prosthetic vacuum pumps.

    Science.gov (United States)

    Komolafe, Oluseeni; Wood, Sean; Caldwell, Ryan; Hansen, Andrew; Fatone, Stefania

    2013-01-01

    Despite increasingly widespread adoption of vacuum-assisted suspension systems in prosthetic clinical practices, there remain gaps in the body of scientific knowledge guiding clinicians' choices of existing products. In this study, we identified important pump-performance metrics and developed techniques to objectively characterize the evacuation performance of prosthetic vacuum pumps. The sensitivity of the proposed techniques was assessed by characterizing the evacuation performance of two electrical (Harmony e-Pulse [Ottobock; Duderstadt, Germany] and LimbLogic VS [Ohio Willow Wood; Mt. Sterling, Ohio]) and three mechanical (Harmony P2, Harmony HD, and Harmony P3 [Ottobock]) prosthetic pumps in bench-top testing. Five fixed volume chambers ranging from 33 cm(3) (2 in.(3)) to 197 cm(3) (12 in.(3)) were used to represent different air volume spaces between a prosthetic socket and a liner-clad residual limb. All measurements were obtained at a vacuum gauge pressure of 57.6 kPa (17 inHg). The proposed techniques demonstrated sensitivity to the different electrical and mechanical pumps and, to a lesser degree, to the different setting adjustments of each pump. The sensitivity was less pronounced for the mechanical pumps, and future improvements for testing of mechanical vacuum pumps were proposed. Overall, this study successfully offers techniques feasible as standards for assessing the evacuation performance of prosthetic vacuum pump devices.

  18. Real-time decision fusion for multimodal neural prosthetic devices.

    Science.gov (United States)

    White, James Robert; Levy, Todd; Bishop, William; Beaty, James D

    2010-03-02

    The field of neural prosthetics aims to develop prosthetic limbs with a brain-computer interface (BCI) through which neural activity is decoded into movements. A natural extension of current research is the incorporation of neural activity from multiple modalities to more accurately estimate the user's intent. The challenge remains how to appropriately combine this information in real-time for a neural prosthetic device. Here we propose a framework based on decision fusion, i.e., fusing predictions from several single-modality decoders to produce a more accurate device state estimate. We examine two algorithms for continuous variable decision fusion: the Kalman filter and artificial neural networks (ANNs). Using simulated cortical neural spike signals, we implemented several successful individual neural decoding algorithms, and tested the capabilities of each fusion method in the context of decoding 2-dimensional endpoint trajectories of a neural prosthetic arm. Extensively testing these methods on random trajectories, we find that on average both the Kalman filter and ANNs successfully fuse the individual decoder estimates to produce more accurate predictions. Our results reveal that a fusion-based approach has the potential to improve prediction accuracy over individual decoders of varying quality, and we hope that this work will encourage multimodal neural prosthetics experiments in the future.

  19. Real-time decision fusion for multimodal neural prosthetic devices.

    Directory of Open Access Journals (Sweden)

    James Robert White

    Full Text Available BACKGROUND: The field of neural prosthetics aims to develop prosthetic limbs with a brain-computer interface (BCI through which neural activity is decoded into movements. A natural extension of current research is the incorporation of neural activity from multiple modalities to more accurately estimate the user's intent. The challenge remains how to appropriately combine this information in real-time for a neural prosthetic device. METHODOLOGY/PRINCIPAL FINDINGS: Here we propose a framework based on decision fusion, i.e., fusing predictions from several single-modality decoders to produce a more accurate device state estimate. We examine two algorithms for continuous variable decision fusion: the Kalman filter and artificial neural networks (ANNs. Using simulated cortical neural spike signals, we implemented several successful individual neural decoding algorithms, and tested the capabilities of each fusion method in the context of decoding 2-dimensional endpoint trajectories of a neural prosthetic arm. Extensively testing these methods on random trajectories, we find that on average both the Kalman filter and ANNs successfully fuse the individual decoder estimates to produce more accurate predictions. CONCLUSIONS: Our results reveal that a fusion-based approach has the potential to improve prediction accuracy over individual decoders of varying quality, and we hope that this work will encourage multimodal neural prosthetics experiments in the future.

  20. Heel-region properties of prosthetic feet and shoes.

    Science.gov (United States)

    Klute, Glenn K; Berge, Jocelyn S; Segal, Ava D

    2004-07-01

    The properties of the prosthetic components prescribed to amputees have the potential to ameliorate or exacerbate their comfort, mobility, and health. To measure the difference in heel-region structural properties of currently available prosthetic feet and shoes, we simulated the period of initial heel-ground contact with a pendulum apparatus. The energy dissipation capacity of the various prosthetic feet ranged from 33.6% to 52.6% of the input energy. Donning a shoe had a large effect. Energy dissipation of a Seattle Lightfoot 2 prosthetic foot was 45.3%, while addition of a walking, running, and orthopedic shoe increased energy dissipation to 63.0%, 73.0%, and 82.4%, respectively. The force versus deformation response to impact was modeled as a hardening spring in parallel with a position-dependent damping element. A nonlinear least-squares curve fit produced model coefficients useful for predicting the heel-region impact response of both prosthetic feet and shoes.

  1. Posttranslational Modification of Heme b in a Bacterial Peroxidase: The Role of Heme to Protein Ester Bonds in Ligand Binding and Catalysis.

    Science.gov (United States)

    Nicolussi, Andrea; Auer, Markus; Weissensteiner, Julia; Schütz, Georg; Katz, Sonja; Maresch, Daniel; Hofbauer, Stefan; Bellei, Marzia; Battistuzzi, Gianantonio; Furtmüller, Paul G; Obinger, Christian

    2017-08-29

    The existence of covalent heme to protein bonds is the most striking structural feature of mammalian peroxidases, including myeloperoxidase and lactoperoxidase (LPO). These autocatalytic posttranslational modifications (PTMs) were shown to strongly influence the biophysical and biochemical properties of these oxidoreductases. Recently, we reported the occurrence of stable LPO-like counterparts with two heme to protein ester linkages in bacteria. This study focuses on the model wild-type peroxidase from the cyanobacterium Lyngbya sp. PCC 8106 (LspPOX) and the mutants D109A, E238A, and D109A/E238A that could be recombinantly produced as apoproteins in Escherichia coli, fully reconstituted to the respective heme b proteins, and posttranslationally modified by hydrogen peroxide. This for the first time allows not only a direct comparison of the catalytic properties of the heme b and PTM forms but also a study of the impact of D109 and E238 on PTM and catalysis, including Compound I formation and the two-electron reduction of Compound I by bromide, iodide, and thiocyanate. It is demonstrated that both heme to protein ester bonds can form independently and that elimination of E238, in contrast to exchange of D109, does not cause significant structural rearrangements or changes in the catalytic properties neither in heme b nor in the PTM form. The obtained findings are discussed with respect to published structural and functional data of human peroxidases.

  2. Direct Tests of Enzymatic Heme Degradation by the Malaria Parasite Plasmodium falciparum*

    Science.gov (United States)

    Sigala, Paul A.; Crowley, Jan R.; Hsieh, Samantha; Henderson, Jeffrey P.; Goldberg, Daniel E.

    2012-01-01

    Malaria parasites generate vast quantities of heme during blood stage infection via hemoglobin digestion and limited de novo biosynthesis, but it remains unclear if parasites metabolize heme for utilization or disposal. Recent in vitro experiments with a heme oxygenase (HO)-like protein from Plasmodium falciparum suggested that parasites may enzymatically degrade some heme to the canonical HO product, biliverdin (BV), or its downstream metabolite, bilirubin (BR). To directly test for BV and BR production by P. falciparum parasites, we DMSO-extracted equal numbers of infected and uninfected erythrocytes and developed a sensitive LC-MS/MS assay to quantify these tetrapyrroles. We found comparable low levels of BV and BR in both samples, suggesting the absence of HO activity in parasites. We further tested live parasites by targeted expression of a fluorescent BV-binding protein within the parasite cytosol, mitochondrion, and plant-like plastid. This probe could detect exogenously added BV but gave no signal indicative of endogenous BV production within parasites. Finally, we recombinantly expressed and tested the proposed heme degrading activity of the HO-like protein, PfHO. Although PfHO bound heme and protoporphyrin IX with modest affinity, it did not catalyze heme degradation in vivo within bacteria or in vitro in UV absorbance and HPLC assays. These observations are consistent with PfHO's lack of a heme-coordinating His residue and suggest an alternative function within parasites. We conclude that P. falciparum parasites lack a canonical HO pathway for heme degradation and thus rely fully on alternative mechanisms for heme detoxification and iron acquisition during blood stage infection. PMID:22992734

  3. Heme Attenuation Ameliorates Irritant Gas Inhalation-Induced Acute Lung Injury.

    Science.gov (United States)

    Aggarwal, Saurabh; Lam, Adam; Bolisetty, Subhashini; Carlisle, Matthew A; Traylor, Amie; Agarwal, Anupam; Matalon, Sadis

    2016-01-10

    Exposure to irritant gases, such as bromine (Br2), poses an environmental and occupational hazard that results in severe lung and systemic injury. However, the mechanism(s) of Br2 toxicity and the therapeutic responses required to mitigate lung damage are not known. Previously, it was demonstrated that Br2 upregulates the heme degrading enzyme, heme oxygenase-1 (HO-1). Since heme is a major inducer of HO-1, we determined whether an increase in heme and heme-dependent oxidative injury underlies the pathogenesis of Br2 toxicity. C57BL/6 mice were exposed to Br2 gas (600 ppm, 30 min) and returned to room air. Thirty minutes postexposure, mice were injected intraperitoneally with a single dose of the heme scavenging protein, hemopexin (Hx) (3 μg/gm body weight), or saline. Twenty-four hours postexposure, saline-treated mice had elevated total heme in bronchoalveolar lavage fluid (BALF) and plasma and acute lung injury (ALI) culminating in 80% mortality after 10 days. Hx treatment significantly lowered heme, decreased evidence of ALI (lower protein and inflammatory cells in BALF, lower lung wet-to-dry weight ratios, and decreased airway hyperreactivity to methacholine), and reduced mortality. In addition, Br2 caused more severe ALI and mortality in mice with HO-1 gene deletion (HO-1-/-) compared to wild-type controls, while transgenic mice overexpressing the human HO-1 gene (hHO-1) showed significant protection. This is the first study delineating the role of heme in ALI caused by Br2. The data suggest that attenuating heme may prove to be a useful adjuvant therapy to treat patients with ALI.

  4. Enhanced heme function and mitochondrial respiration promote the progression of lung cancer cells.

    Science.gov (United States)

    Hooda, Jagmohan; Cadinu, Daniela; Alam, Md Maksudul; Shah, Ajit; Cao, Thai M; Sullivan, Laura A; Brekken, Rolf; Zhang, Li

    2013-01-01

    Lung cancer is the leading cause of cancer-related mortality, and about 85% of the cases are non-small-cell lung cancer (NSCLC). Importantly, recent advance in cancer research suggests that altering cancer cell bioenergetics can provide an effective way to target such advanced cancer cells that have acquired mutations in multiple cellular regulators. This study aims to identify bioenergetic alterations in lung cancer cells by directly measuring and comparing key metabolic activities in a pair of cell lines representing normal and NSCLC cells developed from the same patient. We found that the rates of oxygen consumption and heme biosynthesis were intensified in NSCLC cells. Additionally, the NSCLC cells exhibited substantially increased levels in an array of proteins promoting heme synthesis, uptake and function. These proteins include the rate-limiting heme biosynthetic enzyme ALAS, transporter proteins HRG1 and HCP1 that are involved in heme uptake, and various types of oxygen-utilizing hemoproteins such as cytoglobin and cytochromes. Several types of human tumor xenografts also displayed increased levels of such proteins. Furthermore, we found that lowering heme biosynthesis and uptake, like lowering mitochondrial respiration, effectively reduced oxygen consumption, cancer cell proliferation, migration and colony formation. In contrast, lowering heme degradation does not have an effect on lung cancer cells. These results show that increased heme flux and function are a key feature of NSCLC cells. Further, increased generation and supply of heme and oxygen-utilizing hemoproteins in cancer cells will lead to intensified oxygen consumption and cellular energy production by mitochondrial respiration, which would fuel cancer cell proliferation and progression. The results show that inhibiting heme and respiratory function can effectively arrest the progression of lung cancer cells. Hence, understanding heme function can positively impact on research in lung cancer

  5. Studies of multi-heme cytochromes from Geobacter sulfurreducens

    Energy Technology Data Exchange (ETDEWEB)

    Londer, Yuri; Pokkuluri, P. Raj; Orshonsky, Valerie; Duke, Norma; Schiffer, Marianne

    2004-03-17

    The Geobacteraceae family predominates in the reduction of uranium in subsurface environments. We are focusing on the model organism, Geobacter sulfurreducens; its genome contains a large number (>100) of cytochromes c that function in metal reduction pathways. Intensive functional genomics and physiological studies are in progress in Prof. Derek Lovley's laboratory, and the complete genome sequence of this organism has been determined by Methe et al. 2003. We are studying cytochromes from the c{sub 7} family that are required for the reduction of Fe(III). Previously, we expressed in E. coli (Londer et al., 2002) and determined the three-dimensional structure at 1.45 {angstrom} resolution (Pokkuluri et al., 2004a) of the three-heme cytochrome c{sub 7} (PpcA, coded by ORF01023) characterized by Lloyd et al., 2003. Further we identified in the G. sulfurreducens genome ORFs for several of its homologs (Pokkuluri et al., 2004a). Four of the ORFs are the same size as PpcA; three other ORFs are polymers of c{sub 7}-type domains, two of which consist of four domains and one of nine domains, that contain 12 and 27 hemes respectively.

  6. Disrupted postnatal lung development in heme oxygenase-1 deficient mice

    Directory of Open Access Journals (Sweden)

    Zhang Huayan

    2010-10-01

    Full Text Available Abstract Background Heme oxygenase (HO degrades cellular heme to carbon monoxide, iron and biliverdin. The HO-1 isoform is both inducible and cyto-protective during oxidative stress, inflammation and lung injury. However, little is known about its precise role and function in lung development. We hypothesized that HO-1 is required for mouse postnatal lung alveolar development and that vascular expression of HO-1 is essential and protective during postnatal alveolar development. Methods Neonatal lung development in wildtype and HO-1 mutant mice was evaluated by histological and molecular methods. Furthermore, these newborn mice were treated with postnatal dexamethasone (Dex till postnatal 14 days, and evaluated for lung development. Results Compared to wildtype littermates, HO-1 mutant mice exhibited disrupted lung alveolar structure including simplification, disorganization and reduced secondary crest formation. These defects in alveolar development were more pronounced when these mice were challenged with Dex treatment. Expression levels of both vascular endothelial and alveolar epithelial markers were also further decreased in HO-1 mutants after Dex treatment. Conclusions These experiments demonstrate that HO-1 is required in normal lung development and that HO-1 disruption and dexamethasone exposure are additive in the disruption of postnatal lung growth. We speculate that HO-1 is involved in postnatal lung development through modulation of pulmonary vascular development.

  7. A novel and sensitive assay for heme oxygenase activity.

    Science.gov (United States)

    Iwamori, Saki; Sato, Emiko; Saigusa, Daisuke; Yoshinari, Kouichi; Ito, Sadayoshi; Sato, Hiroshi; Takahashi, Nobuyuki

    2015-10-01

    Heme oxygenase (HO) is a renoprotective protein in the microsome that degrades heme and produces biliverdin. Biliverdin is then reduced to a potent antioxidant bilirubin by biliverdin reductase in the cytosol. Because HO activity does not necessarily correlate with HO mRNA or protein levels, a reliable assay is needed to determine HO activity. Spectrophotometric measurement is tedious and requires a relatively large amount of kidney samples. Moreover, bilirubin is unstable and spontaneously oxidized to biliverdin in vitro. We developed a novel and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify biliverdin to measure HO activity in mice. Biliverdin and its internal standard, a deuterated biliverdin-d4, have MS/MS fragments with m/z transitions of 583 to 297 and 587 to 299, respectively. We prepared lysates of mouse kidneys, and added excess hemin, NADPH, and bilirubin oxidase to convert all bilirubin produced to biliverdin. After 30-min incubation at 37 or 4°C, the samples were analyzed by LC-MS/MS. The difference in the amount of biliverdin between the two temperatures is HO activity. Treating mice with cobalt protoporphyrin, which induces the expression of HO, increased HO activity as determined by biliverdin production. Measuring the production of biliverdin using LC-MS/MS is a more sensitive and specific way to determine HO activity than the spectrophotometric method and allows the detection of subtle changes in renal or other HO activity. Copyright © 2015 the American Physiological Society.

  8. Heme oxygenase-1 comes back to endoplasmic reticulum

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hong Pyo [School of Biological Sciences, Ulsan University (Korea, Republic of); Pae, Hyun-Ock [Department of Immunology, Wonkwang University School of Medicine (Korea, Republic of); Back, Sung Hun; Chung, Su Wol [School of Biological Sciences, Ulsan University (Korea, Republic of); Woo, Je Moon [Department of Opthalmology, Ulasn University Hospital (Korea, Republic of); Son, Yong [Department of Anesthesiology and Pain Medicine, Wonkwang University School of Medicine (Korea, Republic of); Chung, Hun-Taeg, E-mail: chung@ulsan.ac.kr [School of Biological Sciences, Ulsan University (Korea, Republic of)

    2011-01-07

    Research highlights: {yields} Although multiple compartmentalization of HO-1 has been documented, the functional implication of this enzyme at these subcellular organelles is only partially elucidated. {yields} HO-1 expression at ER is induced by a diverse set of conditions that cause ER stressors. {yields} CO may induce HO-1 expression in human ECs by activating Nrf2 through PERK phosphorylation in a positive-feedback manner. {yields} ER-residing HO-1 and its cytoprotective activity against ER stress is discussed. -- Abstract: Originally identified as a rate-limiting enzyme for heme catabolism, heme oxygenase-1 (HO-1) has expanded its roles in anti-inflammation, anti-apoptosis and anti-proliferation for the last decade. Regulation of protein activity by location is well appreciated. Even though multiple compartmentalization of HO-1 has been documented, the functional implication of this enzyme at these subcellular organelles is only partially elucidated. In this review we discuss the endoplasmic reticulum (ER)-residing HO-1 and its cytoprotective activity against ER stress.

  9. Astroglia overexpressing heme oxygenase-1 predispose co-cultured PC12 cells to oxidative injury.

    Science.gov (United States)

    Song, Linyang; Song, Wei; Schipper, Hyman M

    2007-08-01

    The mechanisms responsible for the progressive degeneration of dopaminergic neurons and pathologic iron deposition in the substantia nigra pars compacta of patients with Parkinson's disease (PD) remain unclear. Heme oxygenase-1 (HO-1), the rate-limiting enzyme in the oxidative degradation of heme to ferrous iron, carbon monoxide, and biliverdin, is upregulated in affected PD astroglia and may contribute to abnormal mitochondrial iron sequestration in these cells. To determine whether glial HO-1 hyper-expression is toxic to neuronal compartments, we co-cultured dopaminergic PC12 cells atop monolayers of human (h) HO-1 transfected, sham-transfected, or non-transfected primary rat astroglia. We observed that PC12 cells grown atop hHO-1 transfected astrocytes, but not the astroglia themselves, were significantly more susceptible to dopamine (1 microM) + H(2)O(2) (1 microM)-induced death (assessed by nuclear ethidium monoazide bromide staining and anti-tyrosine hydroxylase immunofluorescence microscopy) relative to control preparations. In the experimental group, PC12 cell death was attenuated significantly by the administration of the HO inhibitor, SnMP (1.5 microM), the antioxidant, ascorbate (200 microM), or the iron chelators, deferoxamine (400 microM), and phenanthroline (100 microM). Exposure to conditioned media derived from HO-1 transfected astrocytes also augmented PC12 cell killing in response to dopamine (1 microM) + H(2)O(2) (1 microM) relative to control media. In PD brain, overexpression of HO-1 in nigral astroglia and accompanying iron liberation may facilitate the bioactivation of dopamine to neurotoxic free radical intermediates and predispose nearby neuronal constituents to oxidative damage. (c) 2007 Wiley-Liss, Inc.

  10. Nanoscale Properties of Neural Cell Prosthetic and Astrocyte Response

    Science.gov (United States)

    Flowers, D. A.; Ayres, V. M.; Delgado-Rivera, R.; Ahmed, I.; Meiners, S. A.

    2009-03-01

    Preliminary data from in-vivo investigations (rat model) suggest that a nanofiber prosthetic device of fibroblast growth factor-2 (FGF-2)-modified nanofibers can correctly guide regenerating axons across an injury gap with aligned functional recovery. Scanning Probe Recognition Microscopy (SPRM) with auto-tracking of individual nanofibers is used for investigation of the key nanoscale properties of the nanofiber prosthetic device for central nervous system tissue engineering and repair. The key properties under SPRM investigation include nanofiber stiffness and surface roughness, nanofiber curvature, nanofiber mesh density and porosity, and growth factor presentation and distribution. Each of these factors has been demonstrated to have global effects on cell morphology, function, proliferation, morphogenesis, migration, and differentiation. The effect of FGF-2 modification on the key nanoscale properties is investigated. Results from the nanofiber prosthetic properties investigations are correlated with astrocyte response to unmodified and FGF-2 modified scaffolds, using 2D planar substrates as a control.

  11. Prosthetic hand control using motion discrimination from EMG signals.

    Science.gov (United States)

    Kurisu, Naoyuki; Tsujiuchi, Nobutaka; Koizumi, Takayuki

    2009-01-01

    In this report, we improve the motion discrimination method from electromyogram (EMG) for a prosthetic hand and propose prosthetic hand control. In the past, we proved that a motion discrimination method using conic models could discriminate three hand motions without the incorrect discriminations that the elbow motions cause. In this research, to increase discrimination accuracy of motion discrimination using conic models, we propose a feature extraction method using quadratic polynomials. Additionally, because many prosthetic hands using motion discrimination have constant motion speed that can't be controlled, we propose an angular velocity generation method using multiple regression models. We verified these methods by controlling the 3D hand model. In the experiment, the proposed method could discriminate five motions at a rate of above 90 percent without the incorrect discriminations that elbow motions cause. Moreover, the wrist joint angle of the 3D hand model could be controlled by standard variation of 3[deg] or less.

  12. Three-Dimensional Printing of Prosthetic Hands for Children.

    Science.gov (United States)

    Burn, Matthew B; Ta, Anderson; Gogola, Gloria R

    2016-05-01

    Children with hand reductions, whether congenital or traumatic, have unique prosthetic needs. They present a challenge because of their continually changing size due to physical growth as well as changing needs due to psychosocial development. Conventional prosthetics are becoming more technologically advanced and increasingly complex. Although these are welcome advances for adults, the concomitant increases in weight, moving parts, and cost are not beneficial for children. Pediatric prosthetic needs may be better met with simpler solutions. Three-dimensional printing can be used to fabricate rugged, light-weight, easily replaceable, and very low cost assistive hands for children. Copyright © 2016 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.

  13. Prosthetic mitral valve thrombosis in pregnancy: from thrombolysis to anticoagulation.

    Science.gov (United States)

    Cardoso, Gonçalo; Aguiar, Carlos; Andrade, Maria João; Patrício, Lino; Freire, Isabel; Serrano, Fátima; Anjos, Rui; Mendes, Miguel

    2015-01-01

    Pregnant women with mechanical prosthetic heart valves are at increased risk for valve thrombosis. Management decisions for this life-threatening complication are complex. Open-heart surgery has a very high risk of maternal mortality and fetal loss. Bleeding and embolic risks associated with thrombolytic agents, the limited efficacy of thrombolysis in certain subgroups, and a lack of experience in the setting of pregnancy raise important concerns. We report a case of mitral prosthetic valve thrombosis in early pregnancy, which was successfully treated with streptokinase. Ten years later, the same patient had an uneventful pregnancy, throughout which acenocoumarol was maintained. With this case we review the prevention (with oral anticoagulant therapy) and treatment of prosthetic valve thrombosis during pregnancy, which is important for both obstetrician and cardiologist. Copyright © 2014 Sociedade Portuguesa de Cardiologia. Published by Elsevier España. All rights reserved.

  14. Surgical-prosthetic treatment of large mandibular cysts

    Directory of Open Access Journals (Sweden)

    Džambas Ljubiša D.

    2003-01-01

    Full Text Available This paper presents a combined surgical-prosthetic procedure of reconstructing mandibular bone defect in a 53 year old patient, following enucleation of a mandibular cyst (Cystectomy Partsch II. After a thorough diagnostic evaluation, a surgical procedure was planned with the particular attention to the nature of the disease, patient’s condition, size and extension of the cyst, tissue loss, and the possibilities of prosthetic management of a mandibular bone defect with partial postresection dental prosthesis. It is of great importance to point to the significance of teamwork of a maxillofacial surgeon and a specialist in prosthodontics. This kind of cooperation provided very effective and less risky soft tissue, as well as bone tissue regeneration (osteogenesis. The patient’s recovery was fast, and he could return to his daily activities and work without significant changes regarding quality of life after surgery and prosthetic treatment.

  15. Suitability of ‘Guidelines for Screening of Prosthetic Candidates: Lower Limb’ for the Eastern Cape, South Africa: A qualitative study

    Directory of Open Access Journals (Sweden)

    Luphiwo L. Mduzana

    2018-01-01

    Full Text Available Background: Major lower limb amputation has a severe impact on functional mobility. Mobility can be salvaged with a prosthesis, but this is not always the best option. It is often difficult to decide whether to refer someone for a prosthesis or not. A prosthetic screening tool ‘Guidelines for Screening of Prosthetic Candidates: Lower Limb’ was developed and is used for prosthetic prescription in parts of the Western Cape province of South Africa.Objectives: This study aimed to explore the suitability of the tool ‘Guidelines for Screening of Prosthetic Candidates: Lower Limb’ for use in the Eastern Cape province of South Africa.Method: A qualitative study was conducted with conveniently sampled occupational therapists (OTs (n = 10, physiotherapists (PTs (n = 12 and prosthetists (n = 6 in government employment in the Buffalo City Metro Municipality. Participants were trained in the use of the tool and used it for four weeks with patients. Their experiences of the tool were assessed through three focus group discussions with emergent themes being identified during inductive data analysis.Findings: Participants indicated that the tool could assist with prosthetic prescription, goal setting, communication and teamwork. They thought that the tool was multidisciplinary in nature, comprehensive and practical. Findings showed a lack of teamwork in this study setting. Resistance to change and a lack of time might also hamper implementation of the tool.Conclusion: The tool can assist with managing the backlog for prostheses and to guide prosthetic prescription in the Eastern Cape province.Clinical implications: A prosthesis can help to salvage functional mobility after lower limb amputation. However, not all people who had above knee amputation manage to walk with a prosthesis. The tool reported on in this article provides information that can guide prosthetic prescription and rehabilitation goals.

  16. Radiological evaluation of prosthetic fit in below-the-knee amputees

    Energy Technology Data Exchange (ETDEWEB)

    Newton, R.L.; Schreiber, M.H.; Morgan, D.

    1988-06-01

    Fourteen artificial limbs in eight adult patients with below the knee amputations were evaluated for prosthetic fit by several radiological parameters. The best objective means of evaluation of prosthetic adequacy is piston action.

  17. Go it alone: four-electron oxidations by mononuclear non-heme iron enzymes.

    Science.gov (United States)

    Peck, Spencer C; van der Donk, Wilfred A

    2017-04-01

    This review discusses the current mechanistic understanding of a group of mononuclear non-heme iron-dependent enzymes that catalyze four-electron oxidation of their organic substrates without the use of any cofactors or cosubstrates. One set of enzymes acts on α-ketoacid-containing substrates, coupling decarboxylation to oxygen activation. This group includes 4-hydroxyphenylpyruvate dioxygenase, 4-hydroxymandelate synthase, and CloR involved in clorobiocin biosynthesis. A second set of enzymes acts on substrates containing a thiol group that coordinates to the iron. This group is comprised of isopenicillin N synthase, thiol dioxygenases, and enzymes involved in the biosynthesis of ergothioneine and ovothiol. The final group of enzymes includes HEPD and MPnS that both carry out the oxidative cleavage of the carbon-carbon bond of 2-hydroxyethylphosphonate but generate different products. Commonalities amongst many of these enzymes are discussed and include the initial substrate oxidation by a ferric-superoxo-intermediate and a second oxidation by a ferryl species.

  18. Maxillofacial prosthetics training and practice profiles in the United States.

    Science.gov (United States)

    Sheets, James L; Yuan, Judy Chia-Chun; Sukotjo, Cortino; Davis, Betsy K; Wee, Alvin G

    2017-10-01

    The motivation of maxillofacial prosthodontists to go into fellowship training and specific procedures in maxillofacial prosthetics practice once they have completed training has not been previously evaluated. The purpose of this study was to survey maxillofacial prosthodontists in the United States to investigate their reasons for pursuing maxillofacial prosthetic training and their practice profiles. In June 2015, a survey was sent to all US maxillofacial prosthodontists asking for descriptive demographics, their reasoning as to what prompted entrance into a maxillofacial prosthetic program, and their practice pattern. Frequencies, percentages, means, and standard deviations were calculated and reported. The survey response rate was 60.4%. The main reason for pursuing maxillofacial training was primarily personal satisfaction, prosthodontic residency exposure, and mentorship rather than media exposure and compensation. The time spent in prosthodontic practice varied among practitioners, with the majority of practice time spent accomplishing standard prosthodontic procedures (65.59%) versus maxillofacial (25.53%) or surgical procedures (9.67%). Of 12 clinical maxillofacial procedures inquired about, the most prevalent were obturators, dental oncology, and mandibular resections. This study reveals that personal satisfaction, mentorship, and prosthodontic residency exposure were the reasons most prosthodontists pursued an additional year of maxillofacial prosthetic fellowship. Most were very satisfied with their training and chosen career path and would recommend an additional year of training. The majority of maxillofacial prosthodontists provided maxillofacial prosthetic treatment for approximately one fourth of their practice time. The most common procedures performed were obturators, dental oncology, and mandibular resections. Copyright © 2017 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

  19. Crystal structure and redox properties of a novel cyanobacterial heme protein with a His/Cys heme axial ligation and a Per-Arnt-Sim (PAS)-like domain.

    Science.gov (United States)

    Motomura, Taiki; Suga, Michihiro; Hienerwadel, Rainer; Nakagawa, Akiko; Lai, Thanh-Lan; Nitschke, Wolfgang; Kuma, Takahiro; Sugiura, Miwa; Boussac, Alain; Shen, Jian-Ren

    2017-06-09

    Photosystem II catalyzes light-induced water oxidation leading to the generation of dioxygen indispensable for sustaining aerobic life on Earth. The Photosystem II reaction center is composed of D1 and D2 proteins encoded by psbA and psbD genes, respectively. In cyanobacteria, different psbA genes are present in the genome. The thermophilic cyanobacterium Thermosynechococcus elongatus contains three psbA genes: psbA1, psbA2, and psbA3, and a new c-type heme protein, Tll0287, was found to be expressed in a strain expressing the psbA2 gene only, but the structure and function of Tll0287 are unknown. Here we solved the crystal structure of Tll0287 at a 2.0 Å resolution. The overall structure of Tll0287 was found to be similar to some kinases and sensor proteins with a Per-Arnt-Sim-like domain rather than to other c-type cytochromes. The fifth and sixth axial ligands for the heme were Cys and His, instead of the His/Met or His/His ligand pairs observed for most of the c-type hemes. The redox potential, E½, of Tll0287 was -255 ± 20 mV versus normal hydrogen electrode at pH values above 7.5. Below this pH value, the E½ increased by ≈57 mV/pH unit at 15 °C, suggesting the involvement of a protonatable group with a pKred = 7.2 ± 0.3. Possible functions of Tll0287 as a redox sensor under microaerobic conditions or a cytochrome subunit of an H2S-oxidizing system are discussed in view of the environmental conditions in which psbA2 is expressed, as well as phylogenetic analysis, structural, and sequence homologies. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Prosthetic Rehabilitation of Ocular Defect resulting from Pediatric Retinoblastoma.

    Science.gov (United States)

    Janya, Suma; Gubrellay, Priyanka; Purwar, Anupam; Khanna, Shally

    2014-01-01

    Ocular defects result from tumor, congenital anomaly and external injury not only lead to serious impairment of function and esthetics but also make the patient psychologically disabled. Prosthetic rehabilitation attempts to restore these disfgurements may improve esthetic, level of function, general psychologic improvement and quality of life. This clinical report details an attempt to rehabilitate a pediatric patient who has undergone orbital enucleation resulting from retinoblastoma with the aid of custom ocular prosthesis using commercially available prefabricated eye shell. How to cite this article: Janya S, Gubrellay P, Purwar A, Khanna S. Prosthetic Rehabilitation of Ocular Defect resulting from Pediatric Retinoblastoma. Int J Clin Pediatr Dent 2014; 7(3):209-212.

  1. Prosthetic limbs on display: from maker to user

    Directory of Open Access Journals (Sweden)

    Sophie Goggins

    2017-11-01

    Full Text Available Prosthetic devices have been used in museums to tell clinical, technical and personal stories. Here we reflect on the ways artificial limbs and their users were represented in recent museum projects at the Royal College of Surgeons of England and at National Museums Scotland. We consider how these meaningful artefacts illuminate three overlapping themes in museum scholarship and practice: the representation of disabled people and disability in museums; reflections on conflict-acquired limb loss; and the presence or otherwise of user or patient voice in interpretation. In working with and representing people who design and wear prosthetics we advocate a balance between narratives of technique and of use.

  2. Towards a Smart Semi-Active Prosthetic Leg: Preliminary Assessment and Testing

    OpenAIRE

    Awad, MI; Abouhossein, A; Dehghani-Sanij, AA; Richardson, R.; Moser, D.; Zahedi, S; Bradley, D.

    2016-01-01

    This paper presents a development of a semi-active prosthetic knee, which can work in both active and passive modes based on the energy required during the gait cycle of various activities of daily livings (ADLs). The prosthetic limb is equipped with various sensors to measure the kinematic and kinetic parameters of both prosthetic limbs. This prosthetic knee is designed to be back-drivable in passive mode to provide a potential use in energy regeneration when there negative energy across the...

  3. Reduced heme levels underlie the exponential growth defect of the Shewanella oneidensis hfq mutant.

    Directory of Open Access Journals (Sweden)

    Christopher M Brennan

    Full Text Available The RNA chaperone Hfq fulfills important roles in small regulatory RNA (sRNA function in many bacteria. Loss of Hfq in the dissimilatory metal reducing bacterium Shewanella oneidensis strain MR-1 results in slow exponential phase growth and a reduced terminal cell density at stationary phase. We have found that the exponential phase growth defect of the hfq mutant in LB is the result of reduced heme levels. Both heme levels and exponential phase growth of the hfq mutant can be completely restored by supplementing LB medium with 5-aminolevulinic acid (5-ALA, the first committed intermediate synthesized during heme synthesis. Increasing expression of gtrA, which encodes the enzyme that catalyzes the first step in heme biosynthesis, also restores heme levels and exponential phase growth of the hfq mutant. Taken together, our data indicate that reduced heme levels are responsible for the exponential growth defect of the S. oneidensis hfq mutant in LB medium and suggest that the S. oneidensis hfq mutant is deficient in heme production at the 5-ALA synthesis step.

  4. Heme Oxygenase, Inflammation, and Fibrosis: The Good, the Bad, and the Ugly?

    Science.gov (United States)

    Lundvig, Ditte M. S.; Immenschuh, Stephan; Wagener, Frank A. D. T. G.

    2012-01-01

    Upon injury, prolonged inflammation and oxidative stress may cause pathological wound healing and fibrosis, leading to formation of excessive scar tissue. Fibrogenesis can occur in most organs and tissues and may ultimately lead to organ dysfunction and failure. The underlying mechanisms of pathological wound healing still remain unclear, and are considered to be multifactorial, but so far, no efficient anti-fibrotic therapies exist. Extra- and intracellular levels of free heme may be increased in a variety of pathological conditions due to release from hemoproteins. Free heme possesses pro-inflammatory and oxidative properties, and may act as a danger signal. Effects of free heme may be counteracted by heme-binding proteins or by heme degradation. Heme is degraded by heme oxygenase (HO) that exists as two isoforms: inducible HO-1 and constitutively expressed HO-2. HO generates the effector molecules biliverdin/bilirubin, carbon monoxide, and free iron/ferritin. HO deficiency in mouse and man leads to exaggerated inflammation following mild insults, and accumulating epidemiological and preclinical studies support the widely recognized notion of the cytoprotective, anti-oxidative, and anti-inflammatory effects of the activity of the HO system and its effector molecules. In this review, we address the potential effects of targeted HO-1 induction or administration of HO-effector molecules as therapeutic targets in fibrotic conditions to counteract inflammatory and oxidative insults. This is exemplified by various clinically relevant conditions, such as hypertrophic scarring, chronic inflammatory liver disease, chronic pancreatitis, and chronic graft rejection in transplantation. PMID:22586396

  5. Regulation of human heme oxygenase-1 gene expression under thermal stress.

    Science.gov (United States)

    Okinaga, S; Takahashi, K; Takeda, K; Yoshizawa, M; Fujita, H; Sasaki, H; Shibahara, S

    1996-06-15

    Heme oxygenase-1 is an essential enzyme in heme catabolism, and its human gene promoter contains a putative heat shock element (HHO-HSE). This study was designed to analyze the regulation of human heme oxygenase-1 gene expression under thermal stress. The amounts of heme oxygenase-1 protein were not increased by heat shock (incubation at 42 degrees C) in human alveolar macrophages and in a human erythroblastic cell line, YN-1-0-A, whereas heat shock protein 70 (HSP70) was noticeably induced. However, heat shock factor does bind in vitro to HHO-HSE and the synthetic HHO-HSE by itself is sufficient to confer the increase in the transient expression of a reporter gene upon heat shock. The deletion of the sequence, located downstream from HHO-HSE, resulted in the activation of a reporter gene by heat shock. These results suggest that HHO-HSE is potentially functional but is repressed in vivo. Interestingly, heat shock abolished the remarkable increase in the levels of heme oxygenase-1 mRNA in YN-1-0-A cells treated with hemin or cadmium, in which HSP70 mRNA was noticeably induced. Furthermore, transient expression assays showed that heat shock inhibits the cadmium-mediated activation of the heme oxygenase-1 promoter, whereas the HSP70 gene promoter was activated upon heat shock. Such regulation of heme oxygenase-1 under thermal stress may be of physiologic significance in erythroid cells.

  6. Regulation of heme metabolism in normal and sideroblastic bone marrow cells in culture

    Energy Technology Data Exchange (ETDEWEB)

    Ibraham, N.G.; Lutton, J.D.; Hoffman, R.; Levere, R.D.

    1985-05-01

    Heme metabolism was examined in developing in vitro erythroid colonies (CFUE) and in bone marrow samples taken directly from four normal donors and four patients with sideroblastic anemia. Maximum activities of delta-aminolevulinic acid synthase (ALAS), ALA dehydratase (ALAD), and /sup 14/C-ALA incorporation into heme were achieved in normal marrow CFUE after 8 days of culture, whereas heme oxygenase progressively decreased to low levels of activity during the same period. Assays on nucleated bone marrow cells taken directly from patients revealed that ALAS activity was considerably reduced in idiopathic sideroblastic anemia (IASA) and X-linked sideroblastic anemia (X-SA) bone marrow specimens, whereas the activity increased more than twofold (normal levels) when cells were assayed from 8-day CFUE. In all cases, ALAD activity appeared to be within normal levels. Measurement of heme synthesis revealed that normal levels of /sup 14/C-ALA incorporation into heme were achieved in IASA cells but were reduced in X-SA cells. In marked contrast to levels in normal cells, heme oxygenase was found to be significantly elevated (two- to fourfold) in bone marrow cells taken directly from patients with IASA and X-SA. Results from this study demonstrate that IASA and X-SA bone marrow cells have disturbances in ALAS and heme metabolism, and that erythropoiesis (CFUE) can be restored to normal levels when cells are cultured in methylcellulose.

  7. Heme-induced Trypanosoma cruzi proliferation is mediated by CaM kinase II

    Energy Technology Data Exchange (ETDEWEB)

    Souza, C.F. [Laboratorio de Imunomodulacao e Protozoologia, Instituto Oswaldo Cruz, Fiocruz (Brazil); Carneiro, A.B.; Silveira, A.B. [Laboratorio de Sinalizacao Celular, Instituto de Bioquimica Medica, UFRJ (Brazil); Laranja, G.A.T. [Laboratorio de Interacao Tripanosomatideos e Vetores, Departamento de Bioquimica, IBRAG, UERJ, 20551-030 Rio de Janeiro (Brazil); Silva-Neto, M.A.C. [Laboratorio de Sinalizacao Celular, Instituto de Bioquimica Medica, UFRJ (Brazil); INCT, Entomologia Molecular (Brazil); Costa, S.C. Goncalves da [Laboratorio de Imunomodulacao e Protozoologia, Instituto Oswaldo Cruz, Fiocruz (Brazil); Paes, M.C., E-mail: mcpaes@uerj.br [Laboratorio de Interacao Tripanosomatideos e Vetores, Departamento de Bioquimica, IBRAG, UERJ, 20551-030 Rio de Janeiro (Brazil); INCT, Entomologia Molecular (Brazil)

    2009-12-18

    Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted through triatomine vectors during their blood-meal on vertebrate hosts. These hematophagous insects usually ingest approximately 10 mM of heme bound to hemoglobin in a single meal. Blood forms of the parasite are transformed into epimastigotes in the crop which initiates a few hours after parasite ingestion. In a previous work, we investigated the role of heme in parasite cell proliferation and showed that the addition of heme significantly increased parasite proliferation in a dose-dependent manner . To investigate whether the heme effect is mediated by protein kinase signalling pathways, parasite proliferation was evaluated in the presence of several protein kinase (PK) inhibitors. We found that only KN-93, a classical inhibitor of calcium-calmodulin-dependent kinases (CaMKs), blocked heme-induced cell proliferation. KN-92, an inactive analogue of KN-93, was not able to block this effect. A T. cruzi CaMKII homologue is most likely the main enzyme involved in this process since parasite proliferation was also blocked when Myr-AIP, an inhibitory peptide for mammalian CaMKII, was included in the cell proliferation assay. Moreover, CaMK activity increased in parasite cells with the addition of heme as shown by immunological and biochemical assays. In conclusion, the present results are the first strong indications that CaMKII is involved in the heme-induced cell signalling pathway that mediates parasite proliferation.

  8. Proinflammatory Responses of Heme in Alveolar Macrophages: Repercussion in Lung Hemorrhagic Episodes

    Directory of Open Access Journals (Sweden)

    Rafael L. Simões

    2013-01-01

    Full Text Available Clinical and experimental observations have supported the notion that free heme released during hemorrhagic and hemolytic episodes may have a major role in lung inflammation. With alveolar macrophages (AM being the main line of defense in lung environments, the influence of free heme on AM activity and function was investigated. We observed that heme in a concentration range found during hemolytic episodes (3–30 μM elicits AM to present a proinflammatory profile, stimulating reactive oxygen species (ROS and nitric oxide (NO generation and inducing IL-1β, IL-6, and IL-10 secretion. ROS production is NADPH oxidase-dependent, being inhibited by DPI and apocynin, and involves p47 subunit phosphorylation. Furthermore, heme induces NF-κB nuclear translocation, iNOS, and also HO-1 expression. Moreover, AM stimulated with free heme show enhanced phagocytic and bactericidal activities. Taken together, the data support a dual role for heme in the inflammatory response associated with lung hemorrhage, acting as a proinflammatory molecule that can either act as both an adjuvant of the innate immunity and as an amplifier of the inflammatory response, leading tissue injury. The understanding of heme effects on pulmonary inflammatory processes can lead to the development of new strategies to ameliorate tissue damage associated with hemorrhagic episodes.

  9. XAS study of the active site of a bacterial heme-sensor

    Energy Technology Data Exchange (ETDEWEB)

    Della Longa, S [Dipartimento di Medicina Sperimentale, Universita dell' Aquila via Vetoio, loc. Coppito II 67100 L' Aquila (Italy); Arcovito, A [Istituto di Biochimica e Biochimica Clinica, Universita Cattolica del Sacro Cuore, Largo F. Vito 1, 00168, Roma (Italy); Brunori, M; Castiglione, N; Cutruzzola, F; Giardina, G; Rinaldo, S [Dipartimento di Scienze Biochimiche ' A. Rossi Fanelli' , Sapienza Universit/a di Roma, P. le A. Moro 5, 00185 Roma (Italy); D' Angelo, P, E-mail: dlonga@caspur.i [Dipartimento di Chimica, Sapienza Universita di Roma, P. le A.Moro 5, 00185 Roma (Italy)

    2009-11-15

    Denitrifying bacteria control NO and NO{sub 2} cytosolic levels by regulating the expression of denitrification gene clusters via REDOX signalling of specific transcriptional factors that may act as NO sensors in vivo. A protein belonging to the subclass DNR (dissimilative nitrate respiration regulator) from Pseudomonas aeruginosa has been recently suggested to be a heme containing protein. Very recently the three dimensional structure of the apo-form of DNR (in the absence of heme) has been determined by X-Ray crystallography, whereas the holo-form (in the presence of heme) has not yet been crystallized. We have investigated the heme local structure in solution of ferric and ferrous holo-DNR by XAS. The Fe K-edge XANES spectrum of the ferric adduct displays typical features of a low-spin hexacoordinate Fe-heme complex, having two histidines ligated. After chemical reduction, relevant changes of the XANES fingerprints suggest a repositioning of the heme inside the hydrophobic core of the protein in agreement with previously reported structural and spectroscopic evidence. Partial release of the axial ligands leaves the Fe(II)heme available, and very reactive, to bind exogenous ligands like NO, thus supporting its role as the cofactor involved in NO sensing activity.

  10. Protein expressed by the ho2 gene of the cyanobacterium Synechocystis sp. PCC 6803 is a true heme oxygenase. Properties of the heme and enzyme complex.

    Science.gov (United States)

    Zhang, Xuhong; Migita, Catharina T; Sato, Michihiko; Sasahara, Masanao; Yoshida, Tadashi

    2005-02-01

    Two isoforms of a heme oxygenase gene, ho1 and ho2, with 51% identity in amino acid sequence have been identified in the cyanobacterium Synechocystis sp. PCC 6803. Isoform-1, Syn HO-1, has been characterized, while isoform-2, Syn HO-2, has not. In this study, a full-length ho2 gene was cloned using synthetic DNA and Syn HO-2 was demonstrated to be highly expressed in Escherichia coli as a soluble, catalytically active protein. Like Syn HO-1, the purified Syn HO-2 bound hemin stoichiometrically to form a heme-enzyme complex and degraded heme to biliverdin IXalpha, CO and iron in the presence of reducing systems such as NADPH/ferredoxin reductase/ferredoxin and sodium ascorbate. The activity of Syn HO-2 was found to be comparable to that of Syn HO-1 by measuring the amount of bilirubin formed. In the reaction with hydrogen peroxide, Syn HO-2 converted heme to verdoheme. This shows that during the conversion of hemin to alpha-meso-hydroxyhemin, hydroperoxo species is the activated oxygen species as in other heme oxygenase reactions. The absorption spectrum of the hemin-Syn HO-2 complex at neutral pH showed a Soret band at 412 nm and two peaks at 540 nm and 575 nm, features observed in the hemin-Syn HO-1 complex at alkaline pH, suggesting that the major species of iron(III) heme iron at neutral pH is a hexa-coordinate low spin species. Electron paramagnetic resonance (EPR) revealed that the iron(III) complex was in dynamic equilibrium between low spin and high spin states, which might be caused by the hydrogen bonding interaction between the distal water ligand and distal helix components. These observations suggest that the structure of the heme pocket of the Syn HO-2 is different from that of Syn HO-1.

  11. The estimated magnitude and direct hospital costs of prosthetic joint infections in the United States, 1997 to 2004.

    Science.gov (United States)

    Hellmann, Matthew; Mehta, Supriya D; Bishai, David M; Mears, Simon C; Zenilman, Jonathan M

    2010-08-01

    To estimate the number and cost of prosthetic joint infection hospitalizations in civilian US hospitals, we analyzed the 1997 to 2004 National Hospital Discharge Survey for the 996.66 International Classification of Diseases discharge code (infection or inflammatory reaction secondary to internal joint prosthesis). The annual number of such hospitalizations averaged 17 589 from 1997 to 2000 and 29 225 from 2001 to 2004. The annual adjusted diagnostic-related group cost for such infection increased from $195 million to $283 million (1997-2004). The mean diagnostic-related group reimbursement ($9034 per hospitalization) did not vary over time or by comorbidity. The nearly doubled number of prosthetic joint infection-related hospitalizations may have been caused by an increased implant rate, changes in patient population, implant procedures, or causative organisms. Copyright (c) 2010 Elsevier Inc. All rights reserved.

  12. Multi-Axis Prosthetic Knee Resembles Alpine Skiing Movements of an Intact Leg.

    Science.gov (United States)

    Demšar, Ivan; Duhovnik, Jože; Lešnik, Blaž; Supej, Matej

    2015-12-01

    The purpose of the study was to analyse the flexion angles of the ski boot, ankle and knee joints of an above-knee prosthesis and to compare them with an intact leg and a control group of skiers. One subject with an above-knee amputation of the right leg and eight healthy subjects simulated the movement of a skiing turn by performing two-leg squats in laboratory conditions. By adding additional loads in proportion to body weight (BW; +1/3 BW, +2/3 BW, +3/3 BW), various skiing regimes were simulated. Change of Flexion Angle (CoFA) and Range of Motion (RoM) in the ski boot, ankle and knee joints were calculated and compared. An average RoM in the skiing boot on the side of prosthesis (4.4 ± 1.1°) was significantly lower compared to an intact leg (5.9 ± 1.8°) and the control group (6.5 ± 2.3°). In the ankle joint, the average RoM was determined to be 13.2±2.9° in the prosthesis, 12.7 ± 2.8° in an intact leg and 14.8±3.6 in the control group. However, the RoM of the knee joint in the prosthesis (42.2 ± 4.2°) was significantly larger than that of the intact leg (34.7 ± 4.4°). The average RoM of the knee joint in the control group was 47.8 ± 5.4°. The influences of additional loads on the kinematics of the lower extremities were different on the side of the prosthesis and on the intact leg. In contrast, additional loads did not produce any significant differences in the control group. Although different CoFAs in the ski boot, ankle and knee joints were used, an above-knee prosthesis with a built-in multi-axis prosthetic knee enables comparable leg kinematics in simulated alpine skiing. Key pointsThe RoM in the ski boot on the side of the prosthetic leg was smaller than the RoM of the intact leg and the control group of healthy subjects.The RoM in the ankle joint of prosthetic leg was comparable to that of the intact leg and the control group of healthy subjects.The RoM in the prosthetic knee joint was greater than the RoM in the knee joint of the intact

  13. Allocation of Heme is Differentially Regulated by Ferrochelatase Isoforms in Arabidopsis Cells

    Directory of Open Access Journals (Sweden)

    Nino Asuela Espinas

    2016-08-01

    Full Text Available Heme is involved in various biological processes as a cofactor of hemoproteins located in various organelles. In plant cells, heme is synthesized by two isoforms of plastid-localized ferrochelatase, FC1 and FC2. In this study, by characterizing Arabidopsis T-DNA insertional mutants, we showed that the allocation of heme is differentially regulated by ferrochelatase isoforms in plant cells. Analyses of weak (fc1-1 and null (fc1-2 mutants suggest that FC1-producing heme is required for initial growth of seedling development. In contrast, weak (fc2-1 and null (fc2-2 mutants of FC2 showed pale green leaves and retarded growth, indicating that FC2-producing heme is necessary for chloroplast development. During the initial growth stage, FC2 deficiency caused reduction of plastid cytochromes. In addition, although FC2 deficiency marginally affected the assembly of photosynthetic reaction center complexes, it caused relatively larger but insufficient light-harvesting antenna to reaction centers, resulting in lower efficiency of photosynthesis. In the later vegetative growth, however, fc2-2 recovered photosynthetic growth, showing that FC1-producing heme may complement the FC2 deficiency. On the other hand, reduced level of cytochromes in microsomal fraction was discovered in fc1-1, suggesting that FC1-producing heme is mainly allocated to extraplastidic organelles. Furthermore, the expression of FC1 is induced by the treatment of an elicitor flg22 while that of FC2 was reduced, and fc1-1 abolished the flg22-dependent induction of FC1 expression and peroxidase activity. Consequently, our results clarified that FC2 produces heme for the photosynthetic machinery in the chloroplast, while FC1 is the housekeeping enzyme providing heme cofactor to the entire cell. In addition, FC1 can partly complement FC2 deficiency and is also involved in defense against stressful conditions.

  14. Proton-directed redox control of O-O bond activation by heme hydroperoxidase models.

    Science.gov (United States)

    Soper, Jake D; Kryatov, Sergey V; Rybak-Akimova, Elena V; Nocera, Daniel G

    2007-04-25

    Hangman metalloporphyrin complexes poise an acid-base group over a redox-active metal center and in doing so allow the "pull" effect of the secondary coordination environment of the heme cofactor of hydroperoxidase enzymes to be modeled. Stopped-flow investigations have been performed to decipher the influence of a proton-donor group on O-O bond activation. Low-temperature reactions of tetramesitylporphyrin (TMP) and Hangman iron complexes containing acid (HPX-CO2H) and methyl ester (HPX-CO2Me) functional groups with peroxyacids generate high-valent Fe=O active sites. Reactions of peroxyacids with (TMP)FeIII(OH) and methyl ester Hangman (HPX-CO2Me)FeIII(OH) give both O-O heterolysis and homolysis products, Compound I (Cpd I) and Compound II (Cpd II), respectively. However, only the former is observed when the hanging group is the acid, (HPX-CO2H)FeIII(OH), because odd-electron homolytic O-O bond cleavage is inhibited. This proton-controlled, 2e- (heterolysis) vs 1e- (homolysis) redox specificity sheds light on the exceptional catalytic performance of the Hangman metalloporphyrin complexes and provides tangible benchmarks for using proton-coupled multielectron reactions to catalyze O-O bond-breaking and bond-making reactions.

  15. Comparison between microprocessor-controlled ankle/foot and conventional prosthetic feet during stair negotiation in people with unilateral transtibial amputation.

    Science.gov (United States)

    Agrawal, Vibhor; Gailey, Robert S; Gaunaurd, Ignacio A; O'Toole, Christopher; Finnieston, Adam A

    2013-01-01

    Contrary to stance-phase dorsiflexion of conventional prosthetic feet, the microprocessor-controlled Proprio foot permits swing-phase dorsiflexion on stairs. The purpose of this study was to compare Symmetry in External Work (SEW) between a microprocessor-controlled foot and conventional prosthetic feet in two groups with unilateral transtibial amputation (Medicare Functional Classification Levels K-Level-2 and K-Level-3) during stair ascent and descent. Ten subjects were evaluated while wearing three conventional prosthetic feet- solid ankle cushion heel (SACH), stationary attachment flexible endoskeleton (SAFE), and Talux-and the Proprio foot using a study socket and were given a 10- to 14-day accommodation period with each foot. Ground reaction forces were collected using F-scan sensors during stair ascent and descent. The SEW between the intact and amputated limbs was calculated for each foot. During stair ascent, the Proprio foot resulted in a higher interlimb symmetry than conventional prosthetic feet, with significant differences between the Pro prio and SACH/SAFE feet. The swing-phase dorsiflexion appeared to promote greater interlimb symmetry because it facilitated forward motion of the body, resulting in a heel-to-toe center of pressure trajectory. During stair descent, all feet had low symmetry without significant differences between feet. The movement strategy used when descending stairs, which is to roll over the edge of a step, had a greater influence on symmetry than the dorsiflexion features of prosthetic feet.

  16. Do distal arteriovenous fistulae improve patency rates of prosthetic infrapopliteal arterial bypasses?

    Science.gov (United States)

    Syrek, J R; Calligaro, K D; Dougherty, M J; Raviola, C A; Rua, I; DeLaurentis, D A

    1998-03-01

    We retrospectively analyzed if distal anastomotic adjunctive arteriovenous fistulae (AVF) improved patency rates of prosthetic bypasses to infrapopliteal arteries. Between July 1, 1991 and June 30, 1996, we performed 43 polytetrafluoroethylene (PTFE) bypasses to infrapopliteal (19 peroneal, 13 anterior tibial, 11 posterior tibial) arteries. All bypasses were performed for limb salvage when autologous vein was not available for a conduit. Adjunctive AVFs were performed in 21 bypasses (PTFE-AVF) and 22 bypasses did not have a fistula (PTFE-ONLY). Patients were allocated to the PTFE-AVF or PTFE-ONLY groups at the discretion of the surgeons, with adjunctive AVFs being performed for small arteries with poor run-off. There were no significant differences in age, sex, site of the proximal anastomosis, or indication for surgery (p > 0.05). There were statistically significant differences in the site of distal anastomosis and quality of arterial run-off based on the Society for Vascular Surgery Ad Hoc Committee on Reporting Standards criteria (p 0.05) and secondary (61% versus 48%) (p > 0.05) patency rates in the PTFE-AVF group versus the PTFE-ONLY group, although limb salvage rates were similar (74% versus 71%) (p > 0.05). Two AVFs required ligation because of steal resulting in diminished distal perfusion. These results support the use of adjunctive distal AVFs to improve overall two-year patency rates of prosthetic infrapopliteal arterial bypasses.

  17. Preoperative Urine Culture Results Correlate Poorly With Bacteriology of Urologic Prosthetic Device Infections.

    Science.gov (United States)

    Kavoussi, Nicholas L; Siegel, Jordan A; Viers, Boyd R; Pagliara, Travis J; Hofer, Matthias D; Cordon, Billy H; Shakir, Nabeel; Scott, Jeremy M; Morey, Allen F

    2017-01-01

    Although preoperative negative urine culture results and treatment of urinary tract infections are generally advised before artificial urinary sphincter (AUS) and penile prosthesis (PP) surgery to prevent device infection, limited evidence exists to support this practice. To evaluate the relation between preoperative urine culture results and the bacteriology of prosthetic device infections. Men undergoing AUS and/or PP placement at a tertiary referral center from 2007 through 2015 were analyzed. A total of 713 devices were implanted in 681 patients (337 AUSs in 314 patients and 376 PPs in 367 patients), of whom 259 (36%) did not have preoperative urine culture and were excluded. The remaining 454 patients received standard broad-spectrum perioperative antibiotics. Two patient groups were identified based on preoperative urine cultures: group 1 had negative urine culture results and group 2 had untreated asymptomatic positive urine culture results identified postoperatively. Device infection was diagnosed clinically and cultures obtained from the explanted device and tissue spaces were compared with preoperative urine culture results. Although multivariate analysis showed that patients undergoing AUS placement had a 4.5-fold greater risk of positive urine culture results (114 of 250, 45%) compared with those undergoing PP placement (36 of 204, 18%; P bacteriology of prosthetic device infections. Copyright © 2016 International Society for Sexual Medicine. Published by Elsevier Inc. All rights reserved.

  18. [Study concerning the influence of the casting parameters of dental alloys on the cast prosthetic rehabilitation].

    Science.gov (United States)

    Viţalariu, Anca Mihaela; Tatarciuc, Monica Silvia

    2008-01-01

    The integration of a cast rehabilitation into the oral environment and its clinical longevity are mainly determined by the accuracy reproduction of the technological parameters. The aim of this study was to evaluate the structural defects induced by the modification of the technological conditions of casting the conventional and titanium alloys. We used 35 cast prosthetic rehabilitation, divided into 7 groups. For each group we modified one parameter: the width of the wax pattern, the design and topography of the casting channels, the absence of the cleanliness of the wax pattern, the preparation of the investment material, the thermal system for the mold preparation and reduced quantity of metal used for casting. For the 7-th group, represented by titanium cast rehabilitation, we modified the design and dimensions of the casting channels. The changes in the technological conditions induced porosity, incomplete frameworks or plus on the prosthetic appliances. The fundamental theoretical knowledge of the clinician and dental technician and the strictly application of the technologies assure the premises of the satisfaction of the patient.

  19. Coordination Chemistry of Linear Oligopyrrolic Fragments Inspired by Heme Metabolites

    Science.gov (United States)

    Gautam, Ritika

    Linear oligopyrroles are degradation products of heme, which is converted in the presence of heme oxygenase to bile pigments, such as biliverdin and bilirubin. These tetrapyrrolic oligopyrroles are ubiquitously present in biological systems and find applications in the fields of catalysis and sensing. These linear tetrapyrrolic scaffolds are further degraded into linear tripyrrolic and dipyrrolic fragments. Although these lower oligopyrroles are abundantly present, their coordination chemistry requires further characterization. This dissertation focuses mainly on two classes of bioinspired linear oligopyrroles, propentdyopent and tripyrrindione, and their transition metal complexes, which present a rich ligand-based redox chemistry. Chapter 1 offers an overview of heme degradation to different classes of linear oligopyrroles and properties of their transition metal complexes. Chapter 2 is focused on the tripyrrin-1,14-dione scaffold of the urinary pigment uroerythrin, which coordinates divalent transition metals palladium and copper with square planar geometry. Specifically, the tripyrrin-1, 14-dione ligand binds Cu(II) and Pd(II) as a dianionic organic radical under ambient conditions. The electrochemical study confirms the presence of ligand based redox chemistry, and one electron oxidation or reduction reactions do not alter the planar geometry around the metal center. The X-Ray analysis and the electron paramagnetic resonance (EPR) studies of the complexes in the solid and solution phase reveals intermolecular interactions between the ligand based unpaired electrons and therefore formation of neutral pi-pi dimers. In Chapter 3, the antioxidant activity and the fluorescence sensor properties of the tripyrrin-1,14-dione ligand in the presence of superoxide are described. We found that the tripyrrindione ligand undergoes one-electron reduction in the presence of the superoxide radical anion (O2•- ) to form highly fluorescent H3TD1•- radical anion, which emits

  20. Heme Oxygenase-1 Promotes Delayed Wound Healing in Diabetic Rats

    Directory of Open Access Journals (Sweden)

    Qing-Ying Chen

    2016-01-01

    Full Text Available Diabetic ulcers are one of the most serious and costly chronic complications for diabetic patients. Hyperglycemia-induced oxidative stress may play an important role in diabetes and its complications. The aim of the study was to explore the effect of heme oxygenase-1 on wound closure in diabetic rats. Diabetic wound model was prepared by making an incision with full thickness in STZ-induced diabetic rats. Wounds from diabetic rats were treated with 10% hemin ointment for 21 days. Increase of HO-1 protein expression enhanced anti-inflammation and antioxidant in diabetic rats. Furthermore, HO-1 increased the levels of VEGF and ICAM-1 and expressions of CBS and CSE protein. In summary, HO-1 promoted the wound closure by augmenting anti-inflammation, antioxidant, and angiogenesis in diabetic rats.

  1. Heme oxygenase system and hypertension: a comprehensive insight.

    Science.gov (United States)

    Tiwari, Shuchita; Ndisang, Joseph Fomusi

    2014-01-01

    Hypertension is a complex interplay of interrelated etiologies, and the leading risk factor for many cardiovascular morbidity and mortality worldwide. Cardinal pathophysiological features of hypertension include enhanced vascular inflammation, vascular remodeling, vascular contractility and increased oxidative stress. In response to oxidative, inflammatory or other noxious stimuli, many physiological pathways like the heme oxygenase (HO) system are activated in an attempt to counteract tissue insults. However, the pathophysiological activation of the HO system only results to a transient increase of HO activity that fall below the necessary threshold capable of activating the downstream signaling components of the HO system like the soluble guanylyl cyclase (sGC)/cyclic guanosine monophosphate (cGMP) secondary messenger system. Therefore, a more robust potentiation of the HO system by pharmacological agents such as hemin, heme-arginate, cobalt protoporphyrin or through retroviral HO-1 gene delivery would be needed to surmount the threshold for cytoprotection. The HO system modulates cellular homeostasis. Importantly, the HO system plays a vital role in a wide spectrum of physiologic including the regulation of blood vessel tone. Alterations in the activity and expression of HO has been correlated to pathophysiology of hypertension and related complications such as hypertrophy, myocardial infarction and heart failure. Moreover, the cytoprotection exerted by HO is attributable to its catabolic products namely, carbon monoxide, bilirubin/biliverdin, and ferritin that are known to modulate immune, inflammatory and oxidative insults. The growing incidence of hypertension and associated cardiometabolic complications has prompted the need for the exploration of alternative therapeutic strategies like substances capable of potentiating the HO system. This review briefly, highlights the functional significance of the HO system and its downstream signaling molecules

  2. Heme oxygenase-1 promoter polymorphisms and risk of spina bifida.

    Science.gov (United States)

    Fujioka, Kazumichi; Yang, Wei; Wallenstein, Matthew B; Zhao, Hui; Wong, Ronald J; Stevenson, David K; Shaw, Gary M

    2015-09-01

    Spina bifida is the most common form of neural tube defects (NTDs). Etiologies of NTDs are multifactorial, and oxidative stress is believed to play a key role in NTD development. Heme oxygenase (HO), the rate-limiting enzyme in heme degradation, has multiple protective properties including mediating antioxidant processes, making it an ideal candidate for study. The inducible HO isoform (HO-1) has two functional genetic polymorphisms: (GT)n dinucleotide repeats and A(-413)T SNP (rs2071746), both of which can affect its promoter activity. However, no study has investigated a possible association between HO-1 genetic polymorphisms and risk of NTDs. This case-control study included 152 spina bifida cases (all myelomeningoceles) and 148 non-malformed controls obtained from the California Birth Defects Monitoring Program reflecting births during 1990 to 1999. Genetic polymorphisms were determined by polymerase chain reaction and amplified fragment length polymorphisms/restriction fragment length polymorphisms using genomic DNA extracted from archived newborn blood spots. Genotype and haplotype frequencies of two HO-1 promoter polymorphisms between cases and controls were compared. For (GT)n dinucleotide repeat lengths and the A(-413)T SNP, no significant differences in allele frequencies or genotypes were found. Linkage disequilibrium was observed between the HO-1 polymorphisms (D': 0.833); however, haplotype analyses did not show increased risk of spina bifida overall or by race/ethnicity. Although, an association was not found between HO-1 polymorphisms and risk of spina bifida, we speculate that the combined effect of low HO-1 expression and exposures to known environmental oxidative stressors (low folate status or diabetes), may overwhelm antioxidant defenses and increase risk of NTDs and warrants further study. © 2015 Wiley Periodicals, Inc.

  3. Rehabilitation regimes based upon psychophysical studies of prosthetic vision

    Science.gov (United States)

    Chen, S. C.; Suaning, G. J.; Morley, J. W.; Lovell, N. H.

    2009-06-01

    Human trials of prototype visual prostheses have successfully elicited visual percepts (phosphenes) in the visual field of implant recipients blinded through retinitis pigmentosa and age-related macular degeneration. Researchers are progressing rapidly towards a device that utilizes individual phosphenes as the elementary building blocks to compose a visual scene. This form of prosthetic vision is expected, in the near term, to have low resolution, large inter-phosphene gaps, distorted spatial distribution of phosphenes, restricted field of view, an eccentrically located phosphene field and limited number of expressible luminance levels. In order to fully realize the potential of these devices, there needs to be a training and rehabilitation program which aims to assist the prosthesis recipients to understand what they are seeing, and also to adapt their viewing habits to optimize the performance of the device. Based on the literature of psychophysical studies in simulated and real prosthetic vision, this paper proposes a comprehensive, theoretical training regime for a prosthesis recipient: visual search, visual acuity, reading, face/object recognition, hand-eye coordination and navigation. The aim of these tasks is to train the recipients to conduct visual scanning, eccentric viewing and reading, discerning low-contrast visual information, and coordinating bodily actions for visual-guided tasks under prosthetic vision. These skills have been identified as playing an important role in making prosthetic vision functional for the daily activities of their recipients.

  4. Different Intraorbital Implant Situations and Ocular Prosthetic Rehabilitation.

    Science.gov (United States)

    Aggarwal, Himanshi; Kumar, Pradeep; Eachempati, Prashanti; Alvi, Habib A

    2016-12-01

    Enucleation is the removal of the entire globe of the eye and a portion of the optic nerve, while evisceration involves the removal of the contents of the globe leaving the sclera, extraocular muscles, and optic nerve. Following enucleation or evisceration, intraorbital implants are routinely placed to enhance the prosthetic outcome in addition to restoring the lost orbital volume. Current practice employs intraorbital implants made of nonporous silicone, hydroxyapatite, or porous polyethylene. Intraorbital implant selection and placement, being a highly demanding procedure in terms of knowledge, skill, and expertise, may be associated with a multiplicity of technical and surgical errors. Complications are usually minimal with these implants, but they do occur. The literature reveals many articles related to intraorbital implants, their benefits, and complications; however, the literature regarding the effect of various intraorbital implant situations on the subsequent prosthetic rehabilitation is markedly scarce. Moreover, the need for interdisciplinary surgical and prosthetic interventions required for successful rehabilitation in cases of compromised implant situations has been underemphasized. Hence, this review aimed to evaluate the effect of different intraorbital implant situations on ocular rehabilitation and the required interdisciplinary surgical and prosthetic treatment approach for rehabilitation of enucleated/eviscerated sockets with compromised implant situations, to provide a critical appraisal, and to present a simplified management strategy. © 2015 by the American College of Prosthodontists.

  5. PROSTHETIC MICROVENOUS GRAFTING IN THE RAT FEMORAL VEIN

    NARCIS (Netherlands)

    VANDERLEI, B; ROBINSON, PH; Bartels, H.

    Male Wistar rats were used to evaluate microvenous prosthetic grafting techniques and microvenous prostheses in the femoral vein. With the end-to-end technique to implant microvenous prostheses, there was extensive exposure of vessel wall collagen especially at the suture sites. Thrombus formation

  6. Prosthetic Hand For Holding Rods, Tools, And Handles

    Science.gov (United States)

    Belcher, Jewell G., Jr.; Vest, Thomas W.

    1995-01-01

    Prosthetic hand with quick-grip/quick-release lever broadens range of specialized functions available to lower-arm amputee by providing improved capabilities for gripping rods, tools, handles, and like. Includes two stationary lower fingers opposed by one pivoting upper finger. Lever operates in conjunction with attached bracket.

  7. Enhanced visual feedback for slip prevention with a prosthetic hand.

    Science.gov (United States)

    Engeberg, Erik D; Meek, Sanford

    2012-12-01

    Upper limb amputees have no direct sense of the grip force applied by a prosthetic hand; thus, precise control of the applied grip force is difficult for amputees. Since there is little object deformation when rigid objects are grasped, it is difficult for amputees to visually gauge the applied grip force in this situation. To determine if the applied grip force from a prosthetic hand can be visually displayed and used to more efficaciously grasp objects. Experimental controlled trial. Force feedback is used in the control algorithm for the prosthetic hand and supplied visually to the user through a bicolor LED experimentally mounted to the thumb. Several experiments are performed by able-bodied test subjects to rate the usefulness of the additional visual feedback when manipulating a clearly visible, brittle object that can break if grasped too firmly. A hybrid force-velocity sliding mode controller is used with and without additional visual force feedback supplied to the operators. Subjective evaluations and success rates from the test subjects indicate a statistically significant reduction in breaking the grasped object when using the prosthesis with the extra visual feedback. The additional visual force feedback can effectively facilitate the manipulation of brittle objects. Clinical relevance The novel approach of this research is the implementation of a noninvasive, effective and economic technique to visually indicate the grip force applied by a prosthetic hand to upper limb amputees. This technique provides a statistically significant improvement when handling brittle objects.

  8. Development of a prototype over-actuated biomimetic prosthetic hand.

    Science.gov (United States)

    Williams, Matthew R; Walter, Wayne

    2015-01-01

    The loss of a hand can greatly affect quality of life. A prosthetic device that can mimic normal hand function is very important to physical and mental recuperation after hand amputation, but the currently available prosthetics do not fully meet the needs of the amputee community. Most prosthetic hands are not dexterous enough to grasp a variety of shaped objects, and those that are tend to be heavy, leading to discomfort while wearing the device. In order to attempt to better simulate human hand function, a dexterous hand was developed that uses an over-actuated mechanism to form grasp shape using intrinsic joint mounted motors in addition to a finger tendon to produce large flexion force for a tight grip. This novel actuation method allows the hand to use small actuators for grip shape formation, and the tendon to produce high grip strength. The hand was capable of producing fingertip flexion force suitable for most activities of daily living. In addition, it was able to produce a range of grasp shapes with natural, independent finger motion, and appearance similar to that of a human hand. The hand also had a mass distribution more similar to a natural forearm and hand compared to contemporary prosthetics due to the more proximal location of the heavier components of the system. This paper describes the design of the hand and controller, as well as the test results.

  9. Prosthetic rehabilitation of a patient with a resected right ...

    African Journals Online (AJOL)

    2015-10-01

    Oct 1, 2015 ... [3] In literature, a mandibular or palatal prosthesis with a guide ramp is one of the methods for reducing mandibular deviation.[1,2,6]. This report describes the implant‑supported prosthetic treatment of a patient with a palatinal guidance ramp‑positioning apparatus after mandibular resection caused by an ...

  10. ORIGINAL ARTICLE Prosthetic management of an 11-year-old ...

    African Journals Online (AJOL)

    Ogunbodede

    Hickey A.J., Vergo T.J. Prothetic treatments for patients with ectodermal dysplasia J. Prosthet. Dent 2001; 86(4):364-368. 13. Bergendral. B. Prothetic habilitation of a young patient with hypohidrotic ectodermal dysplasia and oligodontia: a case report of 20 years of treatment. Int J Prosthodont 2001; 14(5). 471-479. 14.

  11. Consumer satisfaction with the services of prosthetics and orthotics facilities

    NARCIS (Netherlands)

    Bosmans, Joline; Geertzen, Jan; Dijkstra, Pieter U.

    2009-01-01

    Consumer satisfaction with the services provided in a prosthetics and orthotics (PO) facility has seldom been studied. The aim of this study was to analyze consumer satisfaction regarding the services provided by 15 PO facilities in The Netherlands. Consumers (n=1,364) of these PO facilities who

  12. Laser photonics application in prosthetic dentistry for denture design optimization

    Science.gov (United States)

    Grosmann, M. H.; Kiryushin, M. A.; Larkin, A. I.; Lebedenko, A. I.; Lebedenko, I. Yu.; Lopatina, N. A.; Osincev, A. V.; Shchepinov, V. P.; Shchepinova, I. V.

    2010-06-01

    The aim of this work is to demonstrate that holographic and speckle interferometry—laser photonics methods are compatible and useful for prosthetic stomatology. These methods allow to study the deformation of the mandible after insertion of mini-implants of various forms, and to give the practical medical recommendations.

  13. Development of a 3D-Printed Robotic Prosthetic Arm

    Energy Technology Data Exchange (ETDEWEB)

    Gomez Martinez, M.; Garcia-Miquel, A.; Vidal Martinez, N.

    2016-07-01

    Current prostheses are not affordable to the general public. 3D printing technology may allow low-cost production of such devices, making them more readily accessible to people in need. This contribution presents the set-up and the considerations that have to be taken into account to develop a functional artificial upper limb prototype. The robotic prosthetic arm reported herein was produced entirely using 3D printing technology to demonstrate its feasibility on a limited budget. The project was developed to integrate two different functional modes: a prosthetic application and a remote application. The prosthetic application is intended to emulate existing prosthetic devices using myoelectric sensors. The remote application is conceived as a tool for prevention, by providing the general public with a device that could carry out activities that entail a risk of severe physical injury. This is achieved using a hand-tracking system that allows the robotic arm to copy the user’s movements remotely and in real time. The outcome of the validation tests has been considerably successful for both applications and the total costs are on target. (Author)

  14. Prosthetic valve endocarditis after transcatheter aortic valve implantation

    DEFF Research Database (Denmark)

    Olsen, Niels Thue; De Backer, Ole; Thyregod, Hans G H

    2015-01-01

    BACKGROUND: Transcatheter aortic valve implantation (TAVI) is an advancing mode of treatment for inoperable or high-risk patients with aortic stenosis. Prosthetic valve endocarditis (PVE) after TAVI is a serious complication, but only limited data exist on its incidence, outcome, and procedural...

  15. Increased Mortality After Prosthetic Joint Infection in Primary THA

    DEFF Research Database (Denmark)

    Gundtoft, Per Hviid; Pedersen, Alma Becic; Varnum, Claus

    2017-01-01

    Background: Revision for prosthetic joint infection (PJI) has a major effect on patients’ health but it remains unclear if early PJI after primary THA is associated with a high mortality. Questions/Purposes: (1) Do patients with a revision for PJI within 1 year of primary THA have increased...

  16. Specialisation and specialist education in prosthetic dentistry in Europe

    NARCIS (Netherlands)

    Owall, B.; Welfare, R.; Garefis, P.; Hedzelek, W.; Hobkirk, J.; Isidor, F.; Jerolimov, V.; Jokstad, A.; Kalk, W.; Kronstrom, M.; van der Kuij, P.; Mericske-Stern, R.; Naert, I.; Narhi, T.; Nilner, K.; Polyzois, G.; Setz, J.; User, A.; Zonnenberg, A.

    2006-01-01

    This presentation reports on the results of a meeting of prosthodontists from selected European countries. The aim of the meeting was to analyse and promote specialisation and specialist education in Prosthetic Dentistry in Europe. Representatives for Europe were selected from the European

  17. The Design And Development Of Adjustable Prosthetic Device For A ...

    African Journals Online (AJOL)

    Using a six bar link knee mechanism an artificial adjustable leg is produced for an amputee. The prosthetic device when fitted to an amputee provided easy movement for the left leg amputee. It is produced using readily available materials involving aluminum, steel, polyethylene, leader, glue etc. the device has a special ...

  18. Development of a prototype over-actuated biomimetic prosthetic hand.

    Directory of Open Access Journals (Sweden)

    Matthew R Williams

    Full Text Available The loss of a hand can greatly affect quality of life. A prosthetic device that can mimic normal hand function is very important to physical and mental recuperation after hand amputation, but the currently available prosthetics do not fully meet the needs of the amputee community. Most prosthetic hands are not dexterous enough to grasp a variety of shaped objects, and those that are tend to be heavy, leading to discomfort while wearing the device. In order to attempt to better simulate human hand function, a dexterous hand was developed that uses an over-actuated mechanism to form grasp shape using intrinsic joint mounted motors in addition to a finger tendon to produce large flexion force for a tight grip. This novel actuation method allows the hand to use small actuators for grip shape formation, and the tendon to produce high grip strength. The hand was capable of producing fingertip flexion force suitable for most activities of daily living. In addition, it was able to produce a range of grasp shapes with natural, independent finger motion, and appearance similar to that of a human hand. The hand also had a mass distribution more similar to a natural forearm and hand compared to contemporary prosthetics due to the more proximal location of the heavier components of the system. This paper describes the design of the hand and controller, as well as the test results.

  19. Diagnostic flowcharts in osteomyelitis, spondylodiscitis and prosthetic joint infection

    NARCIS (Netherlands)

    Jutte, P.; Lazzeri, E.; Sconfienza, L. M.; Cassar-Pullicino, V.; Trampuz, A.; Petrosillo, N.; Signore, A.

    Infections of the bone, spine and prosthetic joints are serious and complex conditions to diagnose and to treat. Structured diagnostic workup may very well improve the accuracy and speed of diagnosis, thereby improving the outcome since treatment may very well be more successful and less harmful if

  20. Prosthetic prescription in the Netherlands : an observational study

    NARCIS (Netherlands)

    van der Linde, H; Geertzen, JHB; Hofstad, CJ; Postema, K; Van Limbeek, Jacques

    2003-01-01

    Prosthetic prescription for lower limb amputees and the methodology used are primarily based on empirical knowledge. Clinical expertise plays an important role that can lead to an adequate prescription; however, a clear evidence based motivation for the choices made cannot be given. This can lead to

  1. Determining asymmetry of roll-over shapes in prosthetic walking

    NARCIS (Netherlands)

    Curtze, C.; Otten, Bert; Hof, A.L.; Postema, K.

    2011-01-01

    How does the inherent asymmetry of the locomotor system in people with lower-limb amputation affect the ankle-foot roll-over shape of prosthetic walking? In a single-case design, we evaluated the walking patterns of six people with lower-limb amputation (3 transtibial and 3 transfemoral) and three

  2. Renal Haemosiderosis in Patients with· Prosthetic Heart Valves

    African Journals Online (AJOL)

    1974-04-13

    Apr 13, 1974 ... Prosthetic Heart Valves. A. G. ROSE. SUMMARY. Renal haemosiderosis is the anatomical indicator of intra- vascular haemolysis. The incidence of renal haemosiderosis was studied in 66 patients with valve prostheses, 32 patients with advanced rheumatic-type valvular deformities and in 21 consecutive ...

  3. ORIGINAL ARTICLE Prosthetic management of an 11-year-old ...

    African Journals Online (AJOL)

    Ogunbodede

    Result: Upper removable partial denture and lower removable complete denture was fabricated and fitted satisfactory. Conclusion: The psychological and social embarrassment suffered by children with ectodermal dysplasia associated with missing teeth can be greatly improved with early prosthetic rehabilitation with ...

  4. Prosthetic Need between Different Age and Gender ‎With Patient Attending College of Dentistry,Tikrit ‎University

    Directory of Open Access Journals (Sweden)

    Reem Ahmed Shihab‎

    2017-12-01

    Full Text Available Background: the most important factor that affects speak, mastication and aesthetics is tooth loss which in turn imitated on the quality of life. The goal of our research was to evaluate the prosthetic need in sample attending prosthetic department and compare between age and gender. Material and methods: A sample of 244 patients was randomly chosen for questionnaire in the (Removable Prosthodontics Clinic into College of Dentistry-Tikrit University. All subjects were divided in six groups according to their age and also divided according to gender. Intraoral examinations were performed and reasons for extraction were recorded. All statistic calculations were performed using SPSS 23 (Statistical Package for Social Sciences. Results: The total number of 244 patients was involved in this study: 165 (67.6% males and 79 (32.4% females. We found that the higher percentage of patients were in the age group of (20-29 for males which was (23%, 56 patient, while the higher female percentage was found in the age group of (40-49 which was 11.5% . Reasons for extraction recorded the higher percentage due to Caries for both males (75.2% & females (81 % as suspected. Conclusion:The number of patient attending prosthetic department male more than female. Dental caries were the principal cause for extractions in younger patients followed by periodontitis. Society needs more motivation about dental health and care.

  5. [Specificities of prosthetic and orthotic rehabilitation in amputees with head injury].

    Science.gov (United States)

    Teofilovski, Mirko; Parapid, Biljana; Rakić, Miodrag; Popović, Nikola; Teofilovski-Parapid, Gordana

    2011-12-01

    BACKGROUND/AIM. The prosthetic-orthotic rehabilitation (POR) of amputees with head injury within the polytrauma presents a specific entity. The number of traumas caused by the traffic and the low-intensity war conflicts, increases constantly. The aim of our study was to examine the influence of complications on the POR duration and outcome in polytrauma amputees with head injury (PTAHI) recording complications at the beginning and during the POR. The study was carried out on the patients divided into two groups of 35 polytrauma male patients each of corresponding age with unilateral transfemoral amputation caused by the war injury. The experimental group consisted of the amputees with head injury. Standard clinical techniques and procedures, as well as special functional evaluation techniques were used. The PATHI started POR with a greater number of complications (average rate 7.29 vs 5.11 per patient; W = 928.000: Z = 3.730: p = 0.000). There was a highly significant positive correlation between this number and the Barthel Score value change (Fx, H, p admision, the amount of complications defined for the value 4 of POR outcome was significantly higher than values 2 and 3, respectively (H = 8.948; df = 2; p = 0.011). The PTAHI developed significantly more frequently complications during rehabilitation (X2 = 1.061; df = 1; p < 0.01). The proportion of the examinees with the value 4 who developed complications during rehabilitations was significantly higher than those with value 2 (Fp = 3.406; df1 = 2; df2 = 67; p = 0.038). The rehabilitation of the PTAHI lasted significantly longer (average 259.09 vs 183.63 days; W = 923.500; Z = -3.748; p = 0.000). The PTAHI including head injuries started prosthetic-orthotic rehabilitation with more prosthetic complications and their psychological status was worse, resulting in the longer duration of rehabilitation whereas the outcome itself was poor. The value 4 of the prosthetic-orthotic rehabilitation outcome can be expected

  6. Specificities of prosthetic and orthotic rehabilitation in amputees with head injury

    Directory of Open Access Journals (Sweden)

    Teofilovski Mirko

    2011-01-01

    Full Text Available Background/Aim. The prosthetic-orthotic rehabilitation (POR of amputees with head injury within the polytrauma presents a specific entity. The number of traumas caused by the traffic and the low-intensity war conflicts, increases constantly. The aim of our study was to examine the influence of complications on the POR duration and outcome in polytrauma amputees with head injury (PTAHI recording complications at the beginning and during the POR. Methods. The study was carried out on the patients divided into two groups of 35 polytrauma male patients each of corresponding age with unilateral transfemoral amputation caused by the war injury. The experimental group consisted of the amputees with head injury. Standard clinical techniques and procedures, as well as special functional evaluation techniques were used. Results. The PATHI started POR with a greater number of complications (average rate 7.29 vs 5.11 per patient; W = 928.000: Z = 3.730: p = 0.000. There was a highly significant positive correlation between this number and the Barthel Score value change (Fx, H, p < 0.01, and negative significant correlation considering prosthetic use and functional capacity test values (Fx, H p < 0.05. On admision, the amount of complications defined for the value 4 of POR outcome was significantly higher than values 2 and 3, respectively (H = 8.948; df = 2; p = 0.011. The PTAHI developed significantly more frequently complications during rehabilitation (X2 = 1.061; df = 1; p < 0.01. The proportion of the examinees with the value 4 who developed complications during rehabilitations was significantly higher than those with value 2 (Fp = 3.406; df1 = 2; df2 = 67; p = 0.038. The rehabilitation of the PTAHI lasted significantly longer (average 259.09 vs 183.63 days; W = 923.500; Z = -3.748; p = 0.000. Conclusion. The PTAHI including head injuries started prostheticorthotic rehabilitation with more prosthetic complications and their psychological status was worse

  7. Evaluation of 3D printed anatomically scalable transfemoral prosthetic knee.

    Science.gov (United States)

    Ramakrishnan, Tyagi; Schlafly, Millicent; Reed, Kyle B

    2017-07-01

    This case study compares a transfemoral amputee's gait while using the existing Ossur Total Knee 2000 and our novel 3D printed anatomically scalable transfemoral prosthetic knee. The anatomically scalable transfemoral prosthetic knee is 3D printed out of a carbon-fiber and nylon composite that has a gear-mesh coupling with a hard-stop weight-actuated locking mechanism aided by a cross-linked four-bar spring mechanism. This design can be scaled using anatomical dimensions of a human femur and tibia to have a unique fit for each user. The transfemoral amputee who was tested is high functioning and walked on the Computer Assisted Rehabilitation Environment (CAREN) at a self-selected pace. The motion capture and force data that was collected showed that there were distinct differences in the gait dynamics. The data was used to perform the Combined Gait Asymmetry Metric (CGAM), where the scores revealed that the overall asymmetry of the gait on the Ossur Total Knee was more asymmetric than the anatomically scalable transfemoral prosthetic knee. The anatomically scalable transfemoral prosthetic knee had higher peak knee flexion that caused a large step time asymmetry. This made walking on the anatomically scalable transfemoral prosthetic knee more strenuous due to the compensatory movements in adapting to the different dynamics. This can be overcome by tuning the cross-linked spring mechanism to emulate the dynamics of the subject better. The subject stated that the knee would be good for daily use and has the potential to be adapted as a running knee.

  8. A novel method of defective vascular reconstruction using 2-octyl-cyanoacrylate and homemade prosthetic component.

    Science.gov (United States)

    Liang, Xiangdang; Cai, Hongfei; Sun, Geng; Liu, Zihao; Chen, Wen; Tang, Peifu

    2014-01-01

    Currently, adhesive technique is popular in vascular repair but not widely used for defective vessels. This study aimed to determine the feasibility and effectiveness of repairing defective vessels with 2-octyl-cyanoacrylate and a homemade prosthetic component. Homemade prosthetic component consisting of expanded polytetrofluoroethylene (ePTFE), terylene film, and homemade soluble hollow stent mixed with adhesive can replace autologous graft and suture in repairing defective vessels, can fix vessels better using the stent without occlusive bleeding. Forty male mongrel dogs were used, 20 for biomechanical tests and 20 for animal experiments. In the biomechanical test, dogs were randomly divided into two groups (n = 10 each), one group repaired on the two sides of the carotid arteries with 2-octyl-cyanoacrylate and homemade component and another group repaired with suture and ePTFE. Of the 40 specimens, 10 were used for adhesive and 10 for suture specimens for tension strength test, whereas the remaining specimens were used for bursting pressure test. In animal experiments, dogs were also divided into adhesive and suture groups (n = 10), only of the left carotid artery. Recording the operational time, bleeding or not. Vessels were tested using color Doppler ultrasound, the inner diameter was measured, and the degree of stenosis at 8 weeks was evaluated digital subtraction angiography (DSA) were also performed. Specimens were then analyzed histologically. In the adhesive and suture groups, the specimens could afford atension strength of (23.80 ± 1.51) N versus (24.60 ± 1.08) N (P > 0.05), the bursting pressure was (52.03 ± 2.43) kPa versus (50.04 ± 3.51) kPa (P > 0.05), and the mean time of anastomosis was (15.20 ± 0.55) minutes versus (25.97 ± 0.58) minutes (P dog in the adhesive group was bleeding from the suture. One dog from each group presented with thrombosis at 1 week. After measuring using ultrasound, the stenosis degree of all dogs were no more than 30

  9. Cross cultural equivalence testing of the Prosthetic Evaluation Questionnaire (PEQ) for an Arabic speaking population.

    Science.gov (United States)

    Day, Sarah Jane; Buis, Arjan

    2012-06-01

    There are currently no published prosthetic-related outcome measurement tools (OMT) available in the Arabic language. The aim of this study was to translate the Prosthetic Evaluation Questionnaire (PEQ) into Arabic, ensuring cross-cultural equivalence with the original English language version. Psychometric property testing. The PEQ was culturally and linguistically adapted from English to Arabic using a process of forward translation, backward translation, committee review and pre-testing. Pre-testing was carried out in a clinical trial where subjects each completed the questionnaire in Arabic and English, and underwent random probe questioning. The data were compared and analysed, using intraclass correlation (ICC) and Bland Altman plots. Seven patients gave consent and completed the study. For all nine PEQ scales, the ICC point estimate scores were above 0.8, indicating a good degree of correlation. However, for some scales, the 95% confidence interval was wide, indicating a large level of variation. The Bland Altman plots displayed a good distribution around the mean for most of the scales, although the results were affected by the small sample size. The results of the analysis showed that the Arabic version of the PEQ was linguistically equivalent to the original version, although further testing with a larger sample group is recommended. The availability of a prosthetic outcome measurement tool in Arabic will enable clinicians to collect evidence that can be used to monitor and improve patient care. As there is currently little information available about amputees in the Gulf region, this tool will be a useful resource to both clinicians and decision makers.

  10. Replantation versus Prosthetic Fitting in Traumatic Arm Amputations: A Systematic Review.

    Directory of Open Access Journals (Sweden)

    Iris A Otto

    Full Text Available Traumatic arm amputations can be treated with replantation or surgical formalization of the stump with or without subsequent prosthetic fitting. In the literature, many authors suggest the superiority of replantation. This systematic review compared available literature to analyze whether replantation is functionally and psychologically more profitable than formalization and prosthetic fitting in patients with traumatic arm amputation.Functional outcome and satisfaction levels were recorded of patients with amputation levels below elbow, through elbow, and above elbow.Functional outcomes of 301 replantation patients and 172 prosthesis patients were obtained. In the replantation group, good or excellent functional scores were reported in 39% of above elbow, 55% of through elbow, and 50% of below elbow amputation cases. Nearly 100% of patients were satisfied with the replanted limb. In the prosthesis group, full use of the prosthesis was attained in 48% of above elbow and in 89% of below elbow amputation patients. Here, 29% of patients elected not to use the prosthesis for reasons including pain and functional superfluity. In both replantation patients and prosthesis wearers, a below elbow amputation yielded better functional results than higher amputation levels.Replantation of a traumatically amputated arm leads to good function and higher satisfaction rates than a prosthesis, regardless of the objective functional outcome. Sensation and psychological well-being seem the two major advantages of replantation over a prosthesis. The current review of the available literature shows that in carefully selected cases replantation could be the preferred option of treatment.

  11. Replantation versus Prosthetic Fitting in Traumatic Arm Amputations: A Systematic Review.

    Science.gov (United States)

    Otto, Iris A; Kon, Moshe; Schuurman, Arnold H; van Minnen, L Paul

    2015-01-01

    Traumatic arm amputations can be treated with replantation or surgical formalization of the stump with or without subsequent prosthetic fitting. In the literature, many authors suggest the superiority of replantation. This systematic review compared available literature to analyze whether replantation is functionally and psychologically more profitable than formalization and prosthetic fitting in patients with traumatic arm amputation. Functional outcome and satisfaction levels were recorded of patients with amputation levels below elbow, through elbow, and above elbow. Functional outcomes of 301 replantation patients and 172 prosthesis patients were obtained. In the replantation group, good or excellent functional scores were reported in 39% of above elbow, 55% of through elbow, and 50% of below elbow amputation cases. Nearly 100% of patients were satisfied with the replanted limb. In the prosthesis group, full use of the prosthesis was attained in 48% of above elbow and in 89% of below elbow amputation patients. Here, 29% of patients elected not to use the prosthesis for reasons including pain and functional superfluity. In both replantation patients and prosthesis wearers, a below elbow amputation yielded better functional results than higher amputation levels. Replantation of a traumatically amputated arm leads to good function and higher satisfaction rates than a prosthesis, regardless of the objective functional outcome. Sensation and psychological well-being seem the two major advantages of replantation over a prosthesis. The current review of the available literature shows that in carefully selected cases replantation could be the preferred option of treatment.

  12. Dutch evidence-based guidelines for amputation and prosthetics of the lower extremity : Rehabilitation process and prosthetics. Part 2

    NARCIS (Netherlands)

    Geertzen, Jan; van der Linde, Harmen; Rosenbrand, Kitty; Conradi, Marcel; Deckers, Jos; Koning, Jan; Rietman, Hans S.; van der Schaaf, Dick; van der Ploeg, Rein; Schapendonk, Johannes; Schrier, Ernst; Duijzentkunst, Rob Smit; Spruit-van Eijk, Monica; Versteegen, Gerbrig; Voesten, Harrie

    2015-01-01

    Background: A structured, multidisciplinary approach in the rehabilitation process after amputation is needed that includes a greater focus on the involvement of both (para)medics and prosthetists. There is considerable variation in prosthetic prescription concerning the moment of initial prosthesis

  13. Unilateral lower-limb loss: prosthetic device use and functional outcomes in servicemembers from Vietnam war and OIF/OEF conflicts.

    Science.gov (United States)

    Gailey, Robert; McFarland, Lynne V; Cooper, Rory A; Czerniecki, Joseph; Gambel, Jeffrey M; Hubbard, Sharon; Maynard, Charles; Smith, Douglas G; Raya, Michele; Reiber, Gayle E

    2010-01-01

    Rehabilitation goals following major combat-associated limb loss in World War II and the Vietnam war focused on treatment of the injury and a return to civilian life. The goal for Operation Iraqi Freedom/Operation Enduring Freedom (OIF/OEF) servicemembers is to restore function to the greatest possible degree and, if they desire, return them to Active Duty, by providing them with extensive rehabilitation services and a variety of prosthetic devices. Our study determines the usefulness of these diverse types of prosthetic devices for restoring functional capability and documents prosthesis use and satisfaction. We compare servicemembers and veterans with major combat-associated unilateral lower-limb loss: 178 from the Vietnam war and 172 from OIF/OEF conflicts. Of survey participants with unilateral lower-limb loss, 84% of the Vietnam group and 94% of the OIF/OEF group currently use at least one prosthetic device. Reasons for rejection varied by type of device, but common reasons were pain, prosthesis too heavy, and poor fit. Abandonment is infrequent (11% Vietnam group, 4% OIF/OEF group). Future efforts should aim to improve prosthetic-device design, decrease pain, and improve quality of life for these veterans and servicemembers.

  14. The Role of Heme and Reactive Oxygen Species in Proliferation and Survival of Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Marcia Cristina Paes

    2011-01-01

    Full Text Available Trypanosoma cruzi, the protozoan responsible for Chagas disease, has a complex life cycle comprehending two distinct hosts and a series of morphological and functional transformations. Hemoglobin degradation inside the insect vector releases high amounts of heme, and this molecule is known to exert a number of physiological functions. Moreover, the absence of its complete biosynthetic pathway in T. cruzi indicates heme as an essential molecule for this trypanosomatid survival. Within the hosts, T. cruzi has to cope with sudden environmental changes especially in the redox status and heme is able to increase the basal production of reactive oxygen species (ROS which can be also produced as byproducts of the parasite aerobic metabolism. In this regard, ROS sensing is likely to be an important mechanism for the adaptation and interaction of these organisms with their hosts. In this paper we discuss the main features of heme and ROS susceptibility in T. cruzi biology.

  15. Identification and phylogenetic analysis of heme synthesis genes in trypanosomatids and their bacterial endosymbionts.

    Directory of Open Access Journals (Sweden)

    João M P Alves

    Full Text Available It has been known for decades that some insect-infecting trypanosomatids can survive in culture without heme supplementation while others cannot, and that this capability is associated with the presence of a betaproteobacterial endosymbiont in the flagellate's cytoplasm. However, the specific mechanisms involved in this process remained obscure. In this work, we sequence and phylogenetically analyze the heme pathway genes from the symbionts and from their hosts, as well as from a number of heme synthesis-deficient Kinetoplastida. Our results show that the enzymes responsible for synthesis of heme are encoded on the symbiont genomes and produced in close cooperation with the flagellate host. Our evidence suggests that this synergistic relationship is the end result of a history of extensive gene loss and multiple lateral gene transfer events in different branches of the phylogeny of the Trypanosomatidae.

  16. Effects of prosthetic foot forefoot flexibility on gait of unilateral transtibial prosthesis users.

    Science.gov (United States)

    Klodd, Elizabeth; Hansen, Andrew; Fatone, Stefania; Edwards, Mark

    2010-01-01

    Five solid-ankle experimental prosthetic feet were used in this double-blind randomized crossover study to determine the effects of forefoot flexibility on gait of 14 unilateral transtibial prosthesis users. Flexibility in experimental feet was altered by changing the number of flexural hinges in their forefoot sections. When experimental prosthetic foot conditions were compared, measured prosthetic ankle dorsiflexion range of motion increased as much as 3.3° with increasing flexibility (p prosthetic ankle moments as high as 0.53 Nm/kg in late stance phase of walking as flexibility decreased (p prosthetic sides increased as much as 9% of body weight when subjects used the foot with the greatest flexibility (p = 0.001). The results of this study suggest solid-ankle prosthetic foot designs with overly flexible forefoot sections can cause a "drop-off" effect in late stance phase and during the transition of loading between prosthetic and contralateral limbs.

  17. Heme Binding Proteins of Bartonella henselae Are Required when Undergoing Oxidative Stress During Cell and Flea Invasion

    Science.gov (United States)

    Liu, MaFeng; Ferrandez, Yann; Bouhsira, Emilie; Monteil, Martine; Franc, Michel; Boulouis, Henri-Jean; Biville, Francis

    2012-01-01

    Bartonella are hemotropic bacteria responsible for emerging zoonoses. These heme auxotroph alphaproteobacteria must import heme for their growth, since they cannot synthesize it. To import exogenous heme, Bartonella genomes encode for a complete heme uptake system enabling transportation of this compound into the cytoplasm and degrading it to release iron. In addition, these bacteria encode for four or five outer membrane heme binding proteins (Hbps). The structural genes of these highly homologous proteins are expressed differently depending on oxygen, temperature and heme concentrations. These proteins were hypothesized as being involved in various cellular processes according to their ability to bind heme and their regulation profile. In this report, we investigated the roles of the four Hbps of Bartonella henselae, responsible for cat scratch disease. We show that Hbps can bind heme in vitro. They are able to enhance the efficiency of heme uptake when co-expressed with a heme transporter in Escherichia coli. Using B. henselae Hbp knockdown mutants, we show that these proteins are involved in defense against the oxidative stress, colonization of human endothelial cell and survival in the flea. PMID:23144761

  18. Impact of heme on specific antibody production in mice: promotive, inhibitive or null outcome is determined by its concentration

    Directory of Open Access Journals (Sweden)

    Guofu Li

    2017-05-01

    Full Text Available Free heme is an endogenous danger signal that provokes innate immunity. Active innate immunity provides a precondition of an effective adaptive immune response. However, heme catabolites, CO, biliverdin and bilirubin trigger immunosuppression. Furthermore, free heme induces expression of heme oxygenase-1 to increase production of CO, biliverdin and bilirubin. As such, free heme can play a paradoxical role in adaptive immunity. What is the outcome of the animal immune response to an antigen in the presence of free heme? This question remains to be explored. Here, we report the immunization results of rats and mice after intraperitoneal injection of formulations containing BSA and heme. When the heme concentrations were below 1 μM, between 1 μM and 5 μM and above 5 μM, production of anti-BSA IgG and IgM was unaffected, enhanced and suppressed, respectively. The results suggest that heme can influence adaptive immunity by double concentration-thresholds. If the heme concentrations are less than the first threshold, there is no effect on adaptive immunity; if the concentrations are more than the first but less than the second threshold, there is promotion effect; and if the concentrations are more than the second threshold, there is an inhibitory effect. A hypothesis is also presented here to explain the mechanism.

  19. Stability of the prosthetic screws of three types of craniofacial prostheses retention systems.

    Science.gov (United States)

    Lanata-Flores, Antonio Gabriel; Sigua-Rodriguez, Eder Alberto; Goulart, Douglas Rangel; Bomfim-Azevedo, Veber Luiz; Olate, Sergio; de Albergaria-Barbosa, José Ricardo

    2016-12-01

    This study aimed to evaluate the stability of prosthetic screws from three types of craniofacial prostheses retention systems (bar-clip, ball/O-ring, and magnet) when submitted to mechanical cycling. Twelve models of acrylic resin were used with implants placed 20 mm from each other and separated into three groups: (1) bar-clip (Sistema INP, São Paulo, Brazil), (2) ball/O-ring (Sistema INP), and (3) magnet (Metalmag, São Paulo, Brazil), with four samples in each group. Each sample underwent a mechanical cycling removal and insertion test (f=0.5 Hz) to determine the torque and the detorque values of the retention screws. A servo-hydraulic MTS machine (810-Flextest 40; MTS Systems, Eden Prairie, MN, USA) was used to perform the cycling with 2.5 mm and a displacement of 10 mm/s. The screws of the retention systems received an initial torque of 30 Ncm and the torque values required for loosening the screw values were obtained in three cycles (1,080, 2,160, and 3,240). The screws were retorqued to 30 Ncm before each new cycle. The sample was composed of 24 screws grouped as follows: bar-clip (n=8), ball/O-ring (n=8), and magnet (n=8). There were significant differences between the groups, with greater detorque values observed in the ball/O-ring group when compared to the bar-clip and magnet groups for the first cycle. However, the detorque value was greater in the bar-clip group for the second cycle. The results of this study indicate that all prosthetic screws will loosen slightly after an initial tightening torque, also the bar-clip retention system demonstrated greater loosening of the screws when compared with ball/O-ring and magnet retention systems.

  20. Faster heme loss from hemoglobin E than HbS, in acidic pH: Effect ...

    Indian Academy of Sciences (India)

    We report studies on loss of heme at or below pH 3.0 from two clinically important hemoglobin variants, HbE and HbS, in the presence and absence of phopholipid membranes. The kinetics of heme loss has been studied at pH 3.0 to simulate the same at a faster rate than at physiological pH, for spectroscopic investigation.

  1. Heme-Containing Metal-Organic Frameworks for the Oxidative Degradation of Chemical Warfare Agents

    Science.gov (United States)

    2016-04-14

    coordinate cobaltous porphyrin site in a metal – organic framework: structural , EPR, and O, Inorg. Chem. Front., (04 2016): 536. doi: 10.1039...SECURITY CLASSIFICATION OF: This project sought to employ heme-containing metal - organic framework (MOF) materials to carry out the oxidative...15-Apr-2015 14-Jan-2016 Final Report: Heme-Containing Metal - Organic Frameworks for the Oxidative Degradation of Chemical Warfare Agents The views

  2. Heme A synthesis and CcO activity are essential forTrypanosoma cruziinfectivity and replication.

    Science.gov (United States)

    Merli, Marcelo L; Cirulli, Brenda A; Menéndez-Bravo, Simón M; Cricco, Julia A

    2017-06-27

    Trypanosoma cruzi , the causative agent of Chagas disease, presents a complex life cycle and adapts its metabolism to nutrients' availability. Although T. cruzi is an aerobic organism, it does not produce heme. This cofactor is acquired from the host and is distributed and inserted into different heme-proteins such as respiratory complexes in the parasite's mitochondrion. It has been proposed that T. cruzi's energy metabolism relies on a branched respiratory chain with a cytochrome c oxidase-type aa 3 (C c O) as the main terminal oxidase. Heme A, the cofactor for all eukaryotic C c O, is synthesized via two sequential enzymatic reactions catalyzed by heme O synthase (HOS) and heme A synthase (HAS). Previously, TcCox10 and TcCox15 ( Trypanosoma cruzi Cox10 and Cox15 proteins) were identified in T. cruzi They presented HOS and HAS activity, respectively, when they were expressed in yeast. Here, we present the first characterization of TcCox15 in T. cruzi , confirming its role as HAS. It was differentially detected in the different T. cruzi stages, being more abundant in the replicative forms. This regulation could reflect the necessity of more heme A synthesis, and therefore more C c O activity at the replicative stages. Overexpression of a non-functional mutant caused a reduction in heme A content. Moreover, our results clearly showed that this hindrance in the heme A synthesis provoked a reduction on C c O activity and, in consequence, an impairment on T. cruzi survival, proliferation and infectivity. This evidence supports that T. cruzi depends on the respiratory chain activity along its life cycle, being C c O an essential terminal oxidase. © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

  3. Functional analysis and expression of the mono-heme containing cytochrome c subunit of Alternative Complex III in Chloroflexus aurantiacus.

    Science.gov (United States)

    Gao, Xinliu; Majumder, Erica Wunderlich; Kang, Yisheng; Yue, Hai; Blankenship, Robert E

    2013-07-15

    The filamentous anoxygenic phototrophic bacterium Chloroflexus aurantiacus possesses an unusual electron transfer complex called Alternative Complex III instead of the cytochrome bc or bf type complex found in nearly all other known groups of phototrophs. Earlier work has confirmed that Alternative Complex III behaves as a menaquinol:auracyanin oxidoreductase in the photosynthetic electron transfer chain. In this work, we focus on elucidating the contribution of individual subunits to the overall function of Alternative Complex III. The monoheme subunit ActE has been expressed and characterized in Escherichia coli. A partially dissociated Alternative Complex III missing subunit ActE and subunit ActG was obtained by treatment with the chaotropic agent KSCN, and was then reconstituted with the expressed ActE. Enzymatic activity of the partially dissociated Alternative Complex III was greatly reduced and was largely restored in the reconstituted complex. The redox potential of the heme in the recombinant ActE was +385mV vs. NHE, similar to the highest potential heme in the intact complex. The results strongly suggest that the monoheme subunit, ActE, is the terminal electron carrier for Alternative Complex III. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. Simvastatin protects against the development of monocrotaline-induced pulmonary hypertension in rats via a heme oxygenase-1-dependent pathway.

    Science.gov (United States)

    Zhang, Wei-Hua; Zhang, Yun-Jian; Liu, Chun-Ping; Yu, Bing-Xiang; Lu, Wei-Xuan

    2011-10-01

    Heme oxygease-1 (HO-1) is the rate-limiting enzyme in heme catabolism. Induction of HO-1 has been shown to have vasodilatory, anti-inflammatory, and proapoptotic effects. More recently, experimental studies suggested the potential of simvastatin as a novel therapy for pulmonary hypertension (PH); however, the underlying mechanism remains to be investigated. The aim of this study was to evaluate whether HO-1 is required for the pulmonary vascular protective effects of simvastatin. Simvastatin (2 mg/kg/day) was administered once daily to rats for 4 weeks after monocrotaline (MCT) injection. Zn-protoporphyrin (Znpp), a potent inhibitor of HO, was used to confirm the role of HO-1. The hemodynamic changes, right heart hypertrophy, interleukin-6 (IL-6) level, and HO-1 protein expression in lungs were measured at day 28. Simvastatin significantly ameliorated mean pulmonary arterial hypertension (20.6 mm Hg). In addition, perivascular infiltration of inflammatory cells and the level of IL-6 were decreased in simvastatin treatment group. Simvastatin also increased significantly lung HO-1 protein expression. Inhibiting HO-1 using Znpp resulted in a loss of the effect of simvastatin in MCT rats. These results suggest that HO-1 expression is critical for the vascular protective effects of simvastatin in MCT-induced PH rats.

  5. Gastroprotective effect of ghrelin against indomethacin-induced gastric injury in rats: possible role of heme oxygenase-1 pathway.

    Science.gov (United States)

    Allam, Mona M; El-Gohary, Ola A

    2017-07-01

    Ghrelin has been shown to ameliorate gastric injury by several mechanisms in experimental animal models. The present study aimed to investigate the effect of pretreatment with ghrelin on indomethacin-induced gastric injury in rats and the role of heme oxygenase-1(HO-1) pathway as a novel mechanism underlying the gastroprotective effect of ghrelin. In all groups studied, ulcer score (U.S), ulcer index (U.I) and preventive index (P.I) were evaluated and the gastric inflammatory biomarkers including levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and myeloperoxidase (MPO) activity as well as prostaglandin E2 (PGE2), malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), HO-1 and bilirubin as an indicator of heme oxygenase activity were measured. Indomethacin induced significant elevation in U.S and U.I as well as the inflammatory and the oxidative markers and reduced the PGE2 in addition to HO-1 level and activity. Pretreatment with ghrelin reversed these results. In order to elucidate the possible role of HO-1 in mediating the protective effects of ghrelin, tin protoporphyrin (SnPP) HO-1 blocker was administrated; it significantly attenuated the gastroprotective effect of ghrelin. In conclusion HO-1 activity significantly contributes toward ghrelin-mediated gastroprotection.

  6. A mechatronics platform to study prosthetic hand control using EMG signals.

    Science.gov (United States)

    Geethanjali, P

    2016-09-01

    In this paper, a low-cost mechatronics platform for the design and development of robotic hands as well as a surface electromyogram (EMG) pattern recognition system is proposed. This paper also explores various EMG classification techniques using a low-cost electronics system in prosthetic hand applications. The proposed platform involves the development of a four channel EMG signal acquisition system; pattern recognition of acquired EMG signals; and development of a digital controller for a robotic hand. Four-channel surface EMG signals, acquired from ten healthy subjects for six different movements of the hand, were used to analyse pattern recognition in prosthetic hand control. Various time domain features were extracted and grouped into five ensembles to compare the influence of features in feature-selective classifiers (SLR) with widely considered non-feature-selective classifiers, such as neural networks (NN), linear discriminant analysis (LDA) and support vector machines (SVM) applied with different kernels. The results divulged that the average classification accuracy of the SVM, with a linear kernel function, outperforms other classifiers with feature ensembles, Hudgin's feature set and auto regression (AR) coefficients. However, the slight improvement in classification accuracy of SVM incurs more processing time and memory space in the low-level controller. The Kruskal-Wallis (KW) test also shows that there is no significant difference in the classification performance of SLR with Hudgin's feature set to that of SVM with Hudgin's features along with AR coefficients. In addition, the KW test shows that SLR was found to be better in respect to computation time and memory space, which is vital in a low-level controller. Similar to SVM, with a linear kernel function, other non-feature selective LDA and NN classifiers also show a slight improvement in performance using twice the features but with the drawback of increased memory space requirement and time

  7. Prospective comparative study for the evaluation of prosthetic rehabilitation users with transtibial amputation.

    Science.gov (United States)

    Prim, Gabriel de Souza; Santos, Francisco Assis Souza; Vieira, Milton; Nassar, Victor

    2016-10-01

    Individuals with transtibial amputations have difficulties in performing march and stay in balance, directly affecting their quality of life. The use of prostheses can enable the rehabilitation of the individual, but we question how effective are for certain tasks and how they can still improve. To evaluate the prosthesis for transtibial amputation, a comparative study was conducted with two groups: Amputee and NOT Amputees. With the help of Motion Capture technology was held measuring the angles of static balance, walking speed and scores in the execution of daily activities. The results indicate that dispersions of larger static equilibrium angles belonging to the group amputees. In terms of average speed march and in scores of Daily Activities, there was better performance for the group of NOT amputees. From this it was also identified that the technical characteristics of transtibial prosthetic could impact rehabilitation of its members.

  8. Homeostatic response under carcinogen withdrawal, heme oxygenase 1 expression and cell cycle association

    Directory of Open Access Journals (Sweden)

    Batlle Alcira

    2006-12-01

    Full Text Available Abstract Background Chronic injury deregulates cellular homeostasis and induces a number of alterations leading to disruption of cellular processes such as cell cycle checkpoints and apoptosis, driving to carcinogenesis. The stress protein heme oxygenase-1 (HO-1 catalyzes heme degradation producing biliverdin, iron and CO. Induction of HO-1 has been suggested to be essential for a controlled cell growth. The aim of this work was to analyze the in vivo homeostatic response (HR triggered by the withdrawal of a potent carcinogen, p-dimethylaminoazobenzene (DAB, after preneoplastic lesions were observed. We analyzed HO-1 cellular localization and the expression of HO-1, Bcl-2 and cell cycle related proteins under these conditions comparing them to hepatocellular carcinoma (HC. Methods The intoxication protocol was designed based on previous studies demonstrating that preneoplastic lesions were evident after 89 days of chemical carcinogen administration. Male CF1 mice (n = 18 were used. HR group received DAB (0.5 % w/w in the diet for 78 days followed by 11 days of carcinogen deprivation. The HC group received the carcinogen and control animals the standard diet during 89 days. The expression of cell cycle related proteins, of Bcl-2 and of HO-1 were analyzed by western blot. The cellular localization and expression of HO-1 were detected by immnunohistochemistry. Results Increased expression of cyclin E/CDK2 was observed in HR, thus implicating cyclin E/CDK2 in the liver regenerative process. p21cip1/waf1 and Bcl-2 induction in HC was restituted to basal levels in HR. A similar response profile was found for HO-1 expression levels, showing a lower oxidative status in the carcinogen-deprived liver. The immunohistochemical studies revealed the presence of macrophages surrounding foci of necrosis and nodular lesions in HR indicative of an inflammatory response. Furthermore, regenerative cells displayed changes in type, size and intensity of HO-1

  9. Crystal structure of human heme oxygenase-1 in a complex with biliverdin.

    Science.gov (United States)

    Lad, Latesh; Friedman, Jonathan; Li, Huying; Bhaskar, B; Ortiz de Montellano, Paul R; Poulos, Thomas L

    2004-04-06

    Heme oxygenase oxidatively cleaves heme to biliverdin, leading to the release of iron and CO through a process in which the heme participates both as a cofactor and as a substrate. Here we report the crystal structure of the product, iron-free biliverdin, in a complex with human HO-1 at 2.19 A. Structural comparisons of the human biliverdin-HO-1 structure with its heme complex and the recently published rat HO-1 structure in a complex with the biliverdin-iron chelate [Sugishima, M., Sakamoto, H., Higashimoto, Y., Noguchi, M., and Fukuyama, K. (2003) J. Biol. Chem. 278, 32352-32358] show two major differences. First, in the absence of an Fe-His bond and solvent structure in the active site, the distal and proximal helices relax and adopt an "open" conformation which most likely encourages biliverdin release. Second, iron-free biliverdin occupies a different position and orientation relative to heme and the biliverdin-iron complex. Biliverdin adopts a more linear conformation and moves from the heme site to an internal cavity. These structural results provide insight into the rate-limiting step in HO-1 catalysis, which is product, biliverdin, release.

  10. Two Coregulated Efflux Transporters Modulate Intracellular Heme and Protoporphyrin IX Availability in Streptococcus agalactiae

    Science.gov (United States)

    Fernandez, Annabelle; Lechardeur, Delphine; Derré-Bobillot, Aurélie; Couvé, Elisabeth; Gaudu, Philippe; Gruss, Alexandra

    2010-01-01

    Streptococcus agalactiae is a major neonatal pathogen whose infectious route involves septicemia. This pathogen does not synthesize heme, but scavenges it from blood to activate a respiration metabolism, which increases bacterial cell density and is required for full virulence. Factors that regulate heme pools in S. agalactiae are unknown. Here we report that one main strategy of heme and protoporphyrin IX (PPIX) homeostasis in S. agalactiae is based on a regulated system of efflux using two newly characterized operons, gbs1753 gbs1752 (called pefA pefB), and gbs1402 gbs1401 gbs1400 (called pefR pefC pefD), where pef stands for ‘porphyrin-regulated efflux’. In vitro and in vivo data show that PefR, a MarR-superfamily protein, is a repressor of both operons. Heme or PPIX both alleviate PefR-mediated repression. We show that bacteria inactivated for both Pef efflux systems display accrued sensitivity to these porphyrins, and give evidence that they accumulate intracellularly. The ΔpefR mutant, in which both pef operons are up-regulated, is defective for heme-dependent respiration, and attenuated for virulence. We conclude that this new efflux regulon controls intracellular heme and PPIX availability in S. agalactiae, and is needed for its capacity to undergo respiration metabolism, and to infect the host. PMID:20421944

  11. Characterization of Human and Yeast Mitochondrial Glycine Carriers with Implications for Heme Biosynthesis and Anemia.

    Science.gov (United States)

    Lunetti, Paola; Damiano, Fabrizio; De Benedetto, Giuseppe; Siculella, Luisa; Pennetta, Antonio; Muto, Luigina; Paradies, Eleonora; Marobbio, Carlo Marya Thomas; Dolce, Vincenza; Capobianco, Loredana

    2016-09-16

    Heme is an essential molecule in many biological processes, such as transport and storage of oxygen and electron transfer as well as a structural component of hemoproteins. Defects of heme biosynthesis in developing erythroblasts have profound medical implications, as represented by sideroblastic anemia. The synthesis of heme requires the uptake of glycine into the mitochondrial matrix where glycine is condensed with succinyl coenzyme A to yield δ-aminolevulinic acid. Herein we describe the biochemical and molecular characterization of yeast Hem25p and human SLC25A38, providing evidence that they are mitochondrial carriers for glycine. In particular, the hem25Δ mutant manifests a defect in the biosynthesis of δ-aminolevulinic acid and displays reduced levels of downstream heme and mitochondrial cytochromes. The observed defects are rescued by complementation with yeast HEM25 or human SLC25A38 genes. Our results identify new proteins in the heme biosynthetic pathway and demonstrate that Hem25p and its human orthologue SLC25A38 are the main mitochondrial glycine transporters required for heme synthesis, providing definitive evidence of their previously proposed glycine transport function. Furthermore, our work may suggest new therapeutic approaches for the treatment of congenital sideroblastic anemia. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. Two coregulated efflux transporters modulate intracellular heme and protoporphyrin IX availability in Streptococcus agalactiae.

    Directory of Open Access Journals (Sweden)

    Annabelle Fernandez

    2010-04-01

    Full Text Available Streptococcus agalactiae is a major neonatal pathogen whose infectious route involves septicemia. This pathogen does not synthesize heme, but scavenges it from blood to activate a respiration metabolism, which increases bacterial cell density and is required for full virulence. Factors that regulate heme pools in S. agalactiae are unknown. Here we report that one main strategy of heme and protoporphyrin IX (PPIX homeostasis in S. agalactiae is based on a regulated system of efflux using two newly characterized operons, gbs1753 gbs1752 (called pefA pefB, and gbs1402 gbs1401 gbs1400 (called pefR pefC pefD, where pef stands for 'porphyrin-regulated efflux'. In vitro and in vivo data show that PefR, a MarR-superfamily protein, is a repressor of both operons. Heme or PPIX both alleviate PefR-mediated repression. We show that bacteria inactivated for both Pef efflux systems display accrued sensitivity to these porphyrins, and give evidence that they accumulate intracellularly. The DeltapefR mutant, in which both pef operons are up-regulated, is defective for heme-dependent respiration, and attenuated for virulence. We conclude that this new efflux regulon controls intracellular heme and PPIX availability in S. agalactiae, and is needed for its capacity to undergo respiration metabolism, and to infect the host.

  13. Anti-Inflammatory Effect of Angelica gigas via Heme Oxygenase (HO)-1 Expression

    Science.gov (United States)

    Cho, Joon Hyeong; Kwon, Jung Eun; Cho, Youngmi; Kim, Inhye; Kang, Se Chan

    2015-01-01

    Angelica gigas (AG) is effective against various medical conditions such as bacterial infection, inflammation, and cancer. It contains a number of coumarin compounds and the group of interest is the pyranocoumarin, which comprises decursin and decursinol angelate. This group has an effect on controlling inflammation, which is caused by excessive nitric oxide (NO) production. Heme oxygenases (HOs), particularly HO-1, play a role in regulating the production of NO. Thus, this study aimed to investigate the anti-inflammatory effects of AG by measuring HO-1 expression. Treatments with CH2Cl2 layer and Angelica gigas extract (AGE) showed the highest NO inhibition effects. Decursin, decursinol angelate, and nodakenin were isolated from the CH2Cl2 layer of AGE. Decursin also demonstrated the highest anti-oxidative effect among the coumarins. Although decursin had the best NO inhibition and anti-oxidative effects, the effects of AGE treatment far surpassed that of decursin. This is owing to the combination effect of the coumarins present within AGE, which is a solvent extract of AG. The expression of HO-1 is an effective indicator of the anti-inflammatory effects of AG. Based on the results of the coumarin compounds, HO-1 expression was found to be dose dependent and specific to decursin. PMID:26083119

  14. Oral health-related quality of life and prosthetic status of nursing home residents with or without dementia

    Directory of Open Access Journals (Sweden)

    Klotz AL

    2017-04-01

    Full Text Available Anna-Luisa Klotz,1 Alexander Jochen Hassel,1 Johannes Schröder,2,3 Peter Rammelsberg,1 Andreas Zenthöfer1 1Department of Prosthodontics, Dental School, 2Institute of Gerontology, 3Section of Geriatric Psychiatry, University of Heidelberg, Heidelberg, Germany Purpose: The objective of this cross-sectional study was to evaluate the effect of prosthetic status on the oral health-related quality of life (OHRQoL of nursing home residents with or without dementia.Methods: The study was performed in 14 nursing homes across the federal state of Baden-Württemberg, Germany. All eligible participants were included, and general and medical information and information about their dental and prosthetic statuses were collected. The Geriatric Oral Health Assessment Index (GOHAI was administered to evaluate OHRQoL. The Mini-Mental State Examination (MMSE served to classify participants into living or not living with dementia according to the established cutoff value for dementia (MMSE <24. Parametric bivariate statistics and logistic regression models were used to analyze data at P<0.05.Results: A total of 169 participants were included in this study. The mean age of the participants was 82.9 years. A total of some 70% of the sample was living with dementia. The mean GOHAI score along the sample was 49.1 (8.3, and 41% of the sample reported substantially compromised OHRQoL (GOHAI <50. OHRQoL was statistically similar for people with or without dementia (P=0.234; objective oral health was also similar in both groups (P>0.05. The number of teeth (odds ratio [OR]: 2.0, the type of prosthetic status (OR: 6.5, and denture-related treatment needs (OR: 2.4 were the major factors significantly affecting OHRQoL (P<0.05.Conclusion: The OHRQoL of elderly nursing home residents is substantially compromised. Several prosthetic treatment needs for residents living with or without dementia were identified. Edentulism without tooth replacement and having <5 teeth resulted

  15. The Thr-His Connection on the Distal Heme of Catalase-Related Hemoproteins: A Hallmark of Reaction with Fatty Acid Hydroperoxides.

    Science.gov (United States)

    Mashhadi, Zahra; Newcomer, Marcia E; Brash, Alan R

    2016-11-03

    This review focuses on a group of heme peroxidases that retain the catalase fold in structure, yet show little or no reaction with hydrogen peroxide. Instead of having a role in oxidative defense, these enzymes are involved in secondary metabolite biosynthesis. The prototypical enzyme is catalase-related allene oxide synthase, an enzyme that converts a specific fatty acid hydroperoxide to the corresponding allene oxide (epoxide). Other catalase-related enzymes form allylic epoxides, aldehydes, or a bicyclobutane fatty acid. In all catalases (including these relatives), a His residue on the distal face of the heme is absolutely required for activity. Its immediate neighbor in sequence as well as in 3 D space is conserved as Val in true catalases and Thr in the fatty acid hydroperoxide-metabolizing enzymes. Thr-His on the distal face of the heme is critical in switching the substrate specificity from H2 O2 to fatty acid hydroperoxide. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Induction of heme oxygenase-1 protects mouse liver from apoptotic ischemia/reperfusion injury

    Science.gov (United States)

    Issan, Y.; Katz, Y.; Sultan, M.; Safran, M.; Michal, Laniado-Schwartzman; Nader, G. Abraham; Kornowski, R.; Grief, F.; Pappo, O.; Hochhauser, E.

    2017-01-01

    Ischemia/reperfusion (I/R) injury is the main cause of primary graft dysfunction of liver allografts. Cobalt-protoporphyrin (CoPP)–dependent induction of heme oxygenase (HO)-1 has been shown to protect the liver from I/R injury. This study analyzes the apoptotic mechanisms of HO-1-mediated cytoprotection in mouse liver exposed to I/R injury. HO-1 induction was achieved by the administration of CoPP (1.5 mg/kg body weight i.p.). Mice were studied in in vivo model of hepatic segmental (70 %) ischemia for 60 min and reperfusion injury. Mice were randomly allocated to four main experimental groups (n = 10 each): (1) A control group undergoing sham operation. (2) Similar to group 1 but with the administration of CoPP 72 h before the operation. (3) Mice undergoing in vivo hepatic I/R. (4) Similar to group 3 but with the administration of CoPP 72 h before ischemia induction. When compared with the I/R mice group, in the I/R+CoPP mice group, the increased hepatic expression of HO-1 was associated with a significant reduction in liver enzyme levels, fewer apoptotic hepatocytes cells were identified by morphological criteria and by immunohistochemistry for caspase-3, there was a decreased mean number of proliferating cells (positively stained for Ki67), and a reduced hepatic expression of: C/EBP homologous protein (an index of endoplasmic reticulum stress), the NF-κB’s regulated genes (CIAP2, MCP-1 and IL-6), and increased hepatic expression of IκBa (the inhibitory protein of NF-κB). HO-1 over-expression plays a pivotal role in reducing the hepatic apoptotic IR injury. HO-1 may serve as a potential target for therapeutic intervention in hepatic I/R injury during liver transplantation. PMID:23435964

  17. The use of a direct manufacturing prosthetic socket system in a rural community in South Africa: A pilot study and lessons for future research.

    Science.gov (United States)

    Ennion, Liezel; Johannesson, Anton; Rhoda, Anthea

    2017-10-01

    Challenges exist with the provision of appropriate mobility assistive devices in rural areas. The use of the direct manufacturing prosthetic socket system is a possible solution to these challenges. The objective of this study was to test and explore the clients' perspectives with the application of this device. Within a mixed-methods approach, a longitudinal sequential explanatory design was applied. The Orthotic and Prosthetic User's Survey was administered to explore the use of the direct manufacturing prosthetic socket system in terms of function, health-related quality of life and client satisfaction. A conveniently selected sample of 21 individuals who suffered a unilateral trans-tibial amputation was included. Data were collected at 1, 3 and 6 months post fitting, and two focus group discussions were also administered. Of the 21 participants recruited, 11 returned for follow up. Although participants reported favourably about the prosthesis, their scores were generally worse than the norms with regard to function and quality of life. Participants highlighted the need for improvement in the cosmetic appearance of the prosthesis. The direct manufacturing prosthetic socket system could be considered as an alternative technique of socket manufacturing for individuals living in rural areas due to the shorter manufacture time and promising initial results, but further research on this topic with a bigger sample is recommended. Clinical relevance The direct manufacturing prosthetic socket system may be considered as an alternative to the traditional prosthetic socket manufacturing technique used in South Africa. As this device requires only one visit and therefore decreased travel by the patients to the hospitals, it could be applicable to more amputees who cannot return to hospital post discharge.

  18. Prosthetic memory and post-memory: cultural encounters with the past in designing a museum

    Directory of Open Access Journals (Sweden)

    Mălina Ciocea

    2015-07-01

    Full Text Available This paper1 investigates the sources of representations on the communist period and the type of engagement with the past in an experiential museum, in the context of the National Network of Romanian Museums’ project for a laboratory-museum of Romanian Communism. Our analysis of focus-groups in October-November 2012 explores the public’s expectations in terms of museum experience and engagement with objects and the potential of an experiential museum to facilitate deliberation about the past. We use the conceptual framework of recent studies on postmemory (Hirsch, 2008 and prosthetic memory (Landsberg, 2004, 2009 to focus on ways of building the experiential archive needed to produce prosthetic memory. We consider that such an analysis is relevant for two interconnected problems: the bidirectional relationship between a projected museum of communism and a prospective public, and the methodological insights available for investigating this relation. With regard to the first problem, this paper makes a case for treating museums as a memory device rather than a lieu de memoire and analyses the role of the museum in relation to cultural memory. With regard to the second problem, it offers an example of conducting research on prospective publics which departs from traditional marketing approaches, adopting theoretical insights and analytical categories from specific conceptualizations in the field of memory studies.

  19. A CLINICAL STUDY ON THE PERIODONTAL-IMPLANTO-PROSTHETIC REHABILITATION IN PATIENTS WITH PERIODONTAL DISEASES

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    Diana RADU-GHICA

    2012-06-01

    Full Text Available Scope of the study: A comparative analysis on the reliability of the methods of oral rehabilitation of partial edentation through fixed prostheses on dental support vs. prostheses on implants in patients with periodontal diseases. Materials and method: The experimental group was formed of 56 patients (25 men and 31 women, with ages between 23 and 68 years, and different classes of partial edentation in patients with a periodontopathed field. Mention should be made of the fact that, in the case of terminal edentations, a corresponding number of implants has been used, so that to reduce, as much as possible, the number of intermediates involved in the bridge bodies entering the structure of the prosthetic works with dental-implantary support. Results: The survival ratio for the implants substituting the periodontally-induced dental losses was of 90.5%, while that of the implants for the replacement of the teeth lost from other causes (caries, fractures, traumatisms was of 96.5%. Conclusions: The present study supports the assertion that, if the specific conditions of the clinical case under discussion permit it, fixed prosthesizing on a mixed implanto-dental support is more indicated, if considering that this type of prosthesis causes less negative modifications of the prosthetic field, comparatively with the partially-mobile one.

  20. 'Limbitless Solutions': the Prosthetic Arm, Iron Man and the Science Fiction of Technoscience.

    Science.gov (United States)

    Smith, Susan

    2016-12-01

    Early last year, a non-profit organisation called 'Limbitless Solutions' modelled a 3D printed prosthetic arm on a fighting suit that features in the popular superhero film series, Iron Man (2008-2013). In addition, 'Limbitless Solutions' resourcefully deployed the fictional character and inventor of the Iron Man suit, weapons specialist and philanthropist, Tony Stark, played by Robert Downey Jr, in a celebrity/superhero endorsed promotional short film, showing 'Tony', the 'real Iron Man', gifting the futuristic military styled 'gauntlet' to Alex, a 7-year-old boy with a partially developed right arm. Engaging with scholarly work on the science fiction of technoscience, prostheses and the posthuman, and disability and DIY assistive technology, I analyse 'Limbitless Solutions' use of science fiction in a high-profile media event that problematically portrays an impaired child 'in need' of 'repair' and subsequently 'fixed' by technology. Overall, the aim is to integrate science fiction tropes, such as the wounded hero, the fighting suit and prosthetic arm, with disability studies, to highlight the sustained challenges that emerging theories of disability and technology face as contemporary economic, political and ideological forces endorse and promote militarised images of cyborg assimilation over human variation and physical difference. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  1. Minimally invasive prosthetic procedures in the rehabilitation of a bulimic patient affected by dental erosion

    Science.gov (United States)

    Derchi, Giacomo; Peñarrocha, David; Barone, Antonio; Covani, Ugo

    2015-01-01

    The population affected by dental erosion due to bulimia is generally very young. This population group has a high aesthetic requirement; the dentition in these patients is severely damaged, especially in the anterior maxillary quadrant. In terms of treatment, it is still controversial whether an adhesive rehabilitation is preferable to a longer-lasting but more aggressive conventional treatment, such as full-crown coverage of the majority of teeth. This case report describes the prosthetic rehabilitation of a young female patient previously affected by bulimia nervosa and presenting erosion of the maxillary teeth. The prosthetic rehabilitation was performed through indirect adhesive restorations of the anterior teeth and direct restorations of the posterior teeth. A clinical follow-up after 4 years showed that the occlusion remained satisfactorily restored. Posterior direct composite resin restorations and anterior indirect adhesive composite restorations proved to be an effective time and money-saving procedure to rehabilitate patients affected by dental erosion. Adhesive rehabilitation provides a functional and good aesthetic result while preserving tooth structure. Key words:Bulimia, dental erosion, composite resin, veneers. PMID:25810832

  2. Rapid, convenient method for screening imidazole-containing compounds for heme oxygenase inhibition.