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Sample records for avian uncoupling protein

  1. Up-regulation of avian uncoupling protein in cold-acclimated and hyperthyroid ducklings prevents reactive oxygen species production by skeletal muscle mitochondria

    Directory of Open Access Journals (Sweden)

    Servais Stéphane

    2010-04-01

    Full Text Available Abstract Background Although identified in several bird species, the biological role of the avian homolog of mammalian uncoupling proteins (avUCP remains extensively debated. In the present study, the functional properties of isolated mitochondria were examined in physiological or pharmacological situations that induce large changes in avUCP expression in duckling skeletal muscle. Results The abundance of avUCP mRNA, as detected by RT-PCR in gastrocnemius muscle but not in the liver, was markedly increased by cold acclimation (CA or pharmacological hyperthyroidism but was down-regulated by hypothyroidism. Activators of UCPs, such as superoxide with low doses of fatty acids, stimulated a GDP-sensitive proton conductance across the inner membrane of muscle mitochondria from CA or hyperthyroid ducklings. The stimulation was much weaker in controls and not observed in hypothyroid ducklings or in any liver mitochondrial preparations. The production of endogenous mitochondrial reactive oxygen species (ROS was much lower in muscle mitochondria from CA and hyperthyroid ducklings than in the control or hypothyroid groups. The addition of GDP markedly increased the mitochondrial ROS production of CA or hyperthyroid birds up to, or above, the level of control or hypothyroid ducklings. Differences in ROS production among groups could not be attributed to changes in antioxidant enzyme activities (superoxide dismutase or glutathione peroxidase. Conclusion This work provides the first functional in vitro evidence that avian UCP regulates mitochondrial ROS production in situations of enhanced metabolic activity.

  2. Role of Uncoupling Proteins in Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Valle, Adamo; Oliver, Jordi; Roca, Pilar, E-mail: pilar.roca@uib.es [Grupo Multidisciplinar de Oncología Traslacional, Institut Universitari d' Investigació en Ciències de la Salut, Universitat de les Illes Balears/Cra. Valldemossa km 7.5, E-07122, Palma de Mallorca, Illes Balears (Spain)

    2010-04-16

    Uncoupling proteins (UCPs) are a family of inner mitochondrial membrane proteins whose function is to allow the re-entry of protons to the mitochondrial matrix, by dissipating the proton gradient and, subsequently, decreasing membrane potential and production of reactive oxygen species (ROS). Due to their pivotal role in the intersection between energy efficiency and oxidative stress, UCPs are being investigated for a potential role in cancer. In this review we compile the latest evidence showing a link between uncoupling and the carcinogenic process, paying special attention to their involvement in cancer initiation, progression and drug chemoresistance.

  3. Mitochondrial uncoupling proteins and energy metabolism

    Directory of Open Access Journals (Sweden)

    Rosa Anna Busiello

    2015-02-01

    Full Text Available Understanding the metabolic factors that contribute to energy metabolism (EM is critical for the development of new treatments for obesity and related diseases. Mitochondrial oxidative phosphorylation is not perfectly coupled to ATP synthesis, and the process of proton-leak plays a crucial role. Proton-leak accounts for a significant part of the resting metabolic rate and therefore enhancement of this process represents a potential target for obesity treatment. Since their discovery, uncoupling proteins have stimulated great interest due to their involvement in mitochondrial-inducible proton-leak. Despite the widely accepted uncoupling/thermogenic effect of uncoupling protein one (UCP1, which was the first in this family to be discovered, the reactions catalyzed by its homologue UCP3 and the physiological role remain under debate.This review provides an overview of the role played by UCP1 and UCP3 in mitochondrial uncoupling/functionality as well as EM and suggests that they are a potential therapeutic target for treating obesity and its related diseases such as type II diabetes mellitus.

  4. Rapid turnover of mitochondrial uncoupling protein 3

    OpenAIRE

    2010-01-01

    UCP3 (uncoupling protein 3) and its homologues UCP2 and UCP1 are regulators of mitochondrial function. UCP2 is known to have a short half-life of approx. 1 h, owing to its rapid degradation by the cytosolic 26S proteasome, whereas UCP1 is turned over much more slowly by mitochondrial autophagy. In the present study we investigate whether UCP3 also has a short half-life, and whether the proteasome is involved inUCP3 degradation. UCP3 half-life was examined in the mouse C2C12 myoblast cell line...

  5. Uncoupling protein and nonalcoholic fatty liver disease

    Institute of Scientific and Technical Information of China (English)

    JIN Xi; XIANG Zun; CHEN Yi-peng; MA Kui-fen; YE Yue-fang; LI You-ming

    2013-01-01

    Objective To review the current advances on the role of uncoupling protein (UCP) in the pathogenesis and progress of nonalcoholic fatty liver disease (NAFLD).Data sources A comprehensive search of the PubMed literature without restriction on the publication date was carried out using keywords such as UCP and NAFLD.Study selection Articles containing information related to NAFLD and UCP were selected and carefully analyzed.Results The typical concepts,up-to-date findings,and existing controversies of UCP2 in NAFLD were summarized.Besides,the effect of a novel subtype of UCP (hepatocellular down regulated mitochondrial carrier protein,HDMCP) in NAFLD was also analyzed.Finally,the concept that any mitochondrial inner membrane carrier protein may have,more or less,the uncoupling ability was reinforced.Conclusions Considering the importance of NAFLD in clinics and UCP in energy metabolism,we believe that this review may raise research enthusiasm on the effect of UCP in NAFLD and provide a novel mechanism and therapeutic target for NAFLD.

  6. Mitochondrial uncoupling proteins regulate angiotensin-converting enzyme expression

    DEFF Research Database (Denmark)

    Dhamrait, Sukhbir S.; Maubaret, Cecilia; Pedersen-bjergaard, Ulrik

    2016-01-01

    Uncoupling proteins (UCPs) regulate mitochondrial function, and thus cellular metabolism. Angiotensin-converting enzyme (ACE) is the central component of endocrine and local tissue renin–angiotensin systems (RAS), which also regulate diverse aspects of whole-body metabolism and mitochondrial...

  7. Mitochondrial uncoupling proteins regulate angiotensin-converting enzyme expression

    DEFF Research Database (Denmark)

    Dhamrait, Sukhbir S; Maubaret, Cecilia; Pedersen-Bjergaard, Ulrik

    2016-01-01

    Uncoupling proteins (UCPs) regulate mitochondrial function, and thus cellular metabolism. Angiotensin-converting enzyme (ACE) is the central component of endocrine and local tissue renin-angiotensin systems (RAS), which also regulate diverse aspects of whole-body metabolism and mitochondrial...

  8. Molecular cloning of amphioxus uncoupling protein and assessment of its uncoupling activity using a yeast heterologous expression system

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Kun [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China); Sun, Guoxun [Department of Hematology, Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001 (China); Lv, Zhiyuan; Wang, Chen [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China); Jiang, Xueyuan, E-mail: xueyuanjiang@yahoo.com.cn [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China); Li, Donghai, E-mail: lidonghai@gmail.com [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China); Zhang, Chenyu, E-mail: cyzhang@nju.edu.cn [Jiangsu Diabetes Research Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu (China)

    2010-10-01

    Research highlights: {yields} Invertebrates, for example amphioxus, do express uncoupling proteins. {yields} Both the sequence and the uncoupling activity of amphioxus UCP resemble UCP2. {yields} UCP1 is the only UCP that can form dimer on yeast mitochondria. -- Abstract: The present study describes the molecular cloning of a novel cDNA fragment from amphioxus (Branchiostoma belcheri) encoding a 343-amino acid protein that is highly homologous to human uncoupling proteins (UCP), this protein is therefore named amphioxus UCP. This amphioxus UCP shares more homology with and is phylogenetically more related to mammalian UCP2 as compared with UCP1. To further assess the functional similarity of amphioxus UCP to mammalian UCP1 and -2, the amphioxus UCP, rat UCP1, and human UCP2 were separately expressed in Saccharomyces cerevisiae, and the recombinant yeast mitochondria were isolated and assayed for the state 4 respiration rate and proton leak, using pYES2 empty vector as the control. UCP1 increased the state 4 respiration rate by 2.8-fold, and the uncoupling activity was strongly inhibited by GDP, while UCP2 and amphioxus UCP only increased the state 4 respiration rate by 1.5-fold and 1.7-fold in a GDP-insensitive manner, moreover, the proton leak kinetics of amphioxus UCP was very similar to UCP2, but much different from UCP1. In conclusion, the amphioxus UCP has a mild, unregulated uncoupling activity in the yeast system, which resembles mammalian UCP2, but not UCP1.

  9. The Role of Uncoupling Proteins in Diabetes Mellitus

    Directory of Open Access Journals (Sweden)

    Jing Liu

    2013-01-01

    Full Text Available Uncoupling proteins (UCPs are anion carriers expressed in the mitochondrial inner membrane that uncouple oxygen consumption by the respiratory chain from ATP synthesis. The physiological functions of UCPs have long been debated since the new UCPs (UCP2 to 5 were discovered, and the role of UCPs in the pathogeneses of diabetes mellitus is one of the hottest topics. UCPs are thought to be activated by superoxide and then decrease mitochondrial free radicals generation; this may provide a protective effect on diabetes mellitus that is under the oxidative stress conditions. UCP1 is considered to be a candidate gene for diabetes because of its role in thermogenesis and energy expenditure. UCP2 is expressed in several tissues and acts in the negative regulation of insulin secretion by β-cells and in fatty acid metabolism. UCP3 plays a role in fatty acid metabolism and energy homeostasis and modulates insulin sensitivity. Several gene polymorphisms of UCP1, UCP2, and UCP3 were reported to be associated with diabetes. The progress in the role of UCP1, UCP2, and UCP3 on diabetes mellitus is summarized in this review.

  10. Mitochondrial uncoupling proteins in human physiology and disease.

    Science.gov (United States)

    Hagen, T; Vidal-Puig, A

    2002-02-01

    Uncoupling proteins are mitochondrial carrier proteins that catalyse a regulated proton leak across the inner mitochondrial membrane, diverting free energy from ATP synthesis by the mitochondrial F0F1-ATP synthase to the production of heat. Uncoupling protein 1 (UCP1), which is exclusively expressed in brown adipose tissue, is the mediator of thermogenesis in response to beta-adrenergic stimulation. Using gene a knockout mouse model, UCP1 has been shown to be required for cold acclimation. Two homologues of UCP1, UCP2 and UCP3, have been identified recently and show a much wider tissue distribution. UCP2 and UCP3 have been postulated to play a role in the regulation of cold acclimation, energy expenditure and diet-induced thermogenesis in humans, who, in contrast to rodents, have very little brown fat in adult life. However, evidence is accumulating that thermogenesis and regulation of body weight may not be the physiological functions of UCP2 and UCP3. For instance, mice deficient for UCP2 or UCP3 are not cold-intolerant and do not develop obesity. Alternative functions were suggested, primarily based on findings in UCP2 and UCP3 gene knockout mice. Both UCP2- and UCP3-deficient mice were found to overproduce reactive oxygen species and UCP2-deficient mice to hypersecrete insulin. Thus, the UCP1 homologues may play a role in regulating mitochondrial production of reactive oxygen species and b-cell function. In this review, we discuss the role of UCP1, UCP2 and UCP3 in human physiology and disease, primarily based on findings from the various animal models that have been generated.

  11. Retinoids activate proton transport by the uncoupling proteins UCP1 and UCP2.

    OpenAIRE

    1999-01-01

    In mammalian brown adipose tissue, thermogenesis is explained by uncoupling mitochondrial respiration from ATP synthesis. Uncoupling protein-1 (UCP1) is responsible for this uncoupled state, because it allows proton re-entry into the matrix and thus dissipates the proton gradient generated by the respiratory chain. Proton transport by UCP1 is regulated negatively by nucleotides and positively by fatty acids. Adrenergic stimulation of brown adipocytes stimulates lipolysis and therefore enhance...

  12. Protection of chickens against avian hepatitis E virus (avian HEV) infection by immunization with recombinant avian HEV capsid protein.

    Science.gov (United States)

    Guo, H; Zhou, E M; Sun, Z F; Meng, X J

    2007-04-12

    Avian hepatitis E virus (avian HEV) is an emerging virus associated with hepatitis-splenomegaly syndrome in chickens in North America. Avian HEV is genetically and antigenically related to human HEV, the causative agent of hepatitis E in humans. In the lack of a practical animal model, avian HEV infection in chickens has been used as a model to study human HEV replication and pathogenesis. A 32 kDa recombinant ORF2 capsid protein of avian HEV expressed in Escherichia coli was found having similar antigenic structure as that of human HEV containing major neutralizing epitopes. To determine if the capsid protein of avian HEV can be used as a vaccine, 20 chickens were immunized with purified avian HEV recombinant protein with aluminum as adjuvant and another 20 chickens were mock immunized with KLH precipitated in aluminum as controls. Both groups of chickens were subsequently challenged with avian HEV. All the tested mock-immunized control chickens developed typical avian HEV infection characterized by viremia, fecal virus shedding and seroconversion to avian HEV antibodies. Gross hepatic lesions were also found in portion of these chickens. In contrast, none of the tested chickens immunized with avian HEV capsid protein had detectable viremia, fecal virus shedding or observable gross hepatitis lesions. The results from this study suggested that immunization of chickens with avian HEV recombinant ORF2 capsid protein with aluminum as adjuvant can induce protective immunity against avian HEV infection. Chickens are a useful small animal model to study anti-HEV immunity and pathogenesis.

  13. Metabolically inert perfluorinated fatty acids directly activate uncoupling protein 1 in brown-fat mitochondria

    OpenAIRE

    Shabalina, Irina G.; Kalinovich, Anastasia V.; Cannon, Barbara; Nedergaard, Jan

    2015-01-01

    The metabolically inert perfluorinated fatty acids perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) can display fatty acid-like activity in biological systems. The uncoupling protein 1 (UCP1) in brown adipose tissue is physiologically (re)activated by fatty acids, including octanoate. This leads to bioenergetically uncoupled energy dissipation (heat production, thermogenesis). We have examined here the possibility that PFOA/PFOS can directly (re)activate UCP1 in isolated mouse b...

  14. Uncoupling protein expression in skeletal muscle and adipose tissue in response to in vivo porcine somatotropin treatment

    Science.gov (United States)

    The uncoupling proteins are thought to be involved in waste heat production, reducing the energy efficiency of growth in animals. Previous studies have detected their presence in swine and their regulation by the endocrine system. This study attempted to determine whether the uncoupling proteins 2...

  15. A novel amino acid and metabolomics signature in mice overexpressing muscle uncoupling protein 3

    Science.gov (United States)

    Uncoupling protein 3 (UCP3) is highly expressed in skeletal muscle and is known to lower mitochondrial reactive oxygen species and promote fatty acid oxidation; however, the global impact of UCP3 activity on skeletal muscle and whole body metabolism has not been extensively studied. We utilized unt...

  16. Uncoupling Protein 3 Content Is Decreased in Skeletal Muscle of Patients With Type 2 Diabetes

    NARCIS (Netherlands)

    Schrauwen, P.; Hesselink, M.K.C.; Blaak, E.E.; Borghouts, L.B.; Schaart, G.; Saris,; Keizer,

    2001-01-01

    Recently, a role for uncoupling protein-3 (UCP3) in carbohydrate metabolism and in type 2 diabetes has been suggested. Mice overexpressing UCP3 in skeletal muscle showed reduced fasting plasma glucose levels, improved glucose tolerance after an oral glucose load, and reduced fasting plasma insulin l

  17. Glutathionylation Acts as a Control Switch for Uncoupling Proteins UCP2 and UCP3*

    OpenAIRE

    2011-01-01

    The mitochondrial uncoupling proteins 2 and 3 (UCP2 and -3) are known to curtail oxidative stress and participate in a wide array of cellular functions, including insulin secretion and the regulation of satiety. However, the molecular control mechanism(s) governing these proteins remains elusive. Here we reveal that UCP2 and UCP3 contain reactive cysteine residues that can be conjugated to glutathione. We further demonstrate that this modification controls UCP2 and UCP3 function. Both reactiv...

  18. Hydroxynonenal-stimulated activity of the uncoupling protein in Acanthamoeba castellanii mitochondria under phosphorylating conditions.

    Science.gov (United States)

    Woyda-Ploszczyca, Andrzej; Jarmuszkiewicz, Wieslawa

    2013-05-01

    The influence of 4-hydroxy-2-nonenal (HNE), a lipid peroxidation end product, on the activity of the amoeba Acanthamoeba castellanii uncoupling protein (AcUCP) in isolated phosphorylating mitochondria was studied. Under phosphorylating conditions, exogenously added HNE induced GTP-sensitive AcUCP-mediated mitochondrial uncoupling. The HNE-induced proton leak decreased the yield of oxidative phosphorylation in an HNE concentration-dependent manner. The present study describes how the contributions of ATP synthase and HNE-induced AcUCP in phosphorylating respiration vary when the rate of succinate oxidation is decreased by limiting succinate uptake or inhibiting complex III activity within the range of a constant membrane potential. In phosphorylating mitochondria, at a given HNE concentration (100 μM), the efficiency of AcUCP in mitochondrial uncoupling increased as the respiratory rate decreased because the AcUCP contribution remained constant while the ATP synthase contribution decreased with the respiratory rate. HNE-induced uncoupling can be inhibited by GTP only when ubiquinone is sufficiently oxidized, indicating that in phosphorylating A. castellanii mitochondria, the sensitivity of AcUCP activity to GTP depends on the redox state of the membranous ubiquinone.

  19. Role of mitochondrial uncoupling protein 2 in cancer cell resistance to gemcitabine.

    Science.gov (United States)

    Dalla Pozza, Elisa; Fiorini, Claudia; Dando, Ilaria; Menegazzi, Marta; Sgarbossa, Anna; Costanzo, Chiara; Palmieri, Marta; Donadelli, Massimo

    2012-10-01

    Cancer cells exhibit an endogenous constitutive oxidative stress higher than that of normal cells, which renders tumours vulnerable to further reactive oxygen species (ROS) production. Mitochondrial uncoupling protein 2 (UCP2) can mitigate oxidative stress by increasing the influx of protons into the mitochondrial matrix and reducing electron leakage and mitochondrial superoxide generation. Here, we demonstrate that chemical uncouplers or UCP2 over-expression strongly decrease mitochondrial superoxide induction by the anticancer drug gemcitabine (GEM) and protect cancer cells from GEM-induced apoptosis. Moreover, we show that GEM IC(50) values well correlate with the endogenous level of UCP2 mRNA, suggesting a critical role for mitochondrial uncoupling in GEM resistance. Interestingly, GEM treatment stimulates UCP2 mRNA expression suggesting that mitochondrial uncoupling could have a role also in the acquired resistance to GEM. Conversely, UCP2 inhibition by genipin or UCP2 mRNA silencing strongly enhances GEM-induced mitochondrial superoxide generation and apoptosis, synergistically inhibiting cancer cell proliferation. These events are significantly reduced by the addition of the radical scavenger N-acetyl-l-cysteine or MnSOD over-expression, demonstrating a critical role of the oxidative stress. Normal primary fibroblasts are much less sensitive to GEM/genipin combination. Our results demonstrate for the first time that UCP2 has a role in cancer cell resistance to GEM supporting the development of an anti-cancer therapy based on UCP2 inhibition associated to GEM treatment.

  20. Uncoupling in Secondary Transport Proteins. A Mechanistic Explanation for Mutants of lac Permease with an Uncoupled Phenotype

    NARCIS (Netherlands)

    Lolkema, J.S.; Poolman, B.

    1995-01-01

    The kinetic behavior of a H+-substrate symporter has been studied in which in addition to the unloaded (E) and fully loaded states (E.S.H) of the carrier also one of the binary complexes (E.S or E.H) may reorient its binding sites. This results in two types of uncoupled mutants, the ES leak and the

  1. The emerging neuroprotective role of mitochondrial uncoupling protein-2 in traumatic brain injury

    OpenAIRE

    2015-01-01

    Traumatic brain injury (TBI) is a multifaceted disease with intrinsically complex heterogeneity and remains a significant clinical challenge to manage. TBI model systems have demonstrated many mechanisms that contribute to brain parenchymal cell death, including glutamate and calcium toxicity, oxidative stress, inflammation, and mitochondrial dysfunction. Mitochondria are critically regulated by uncoupling proteins (UCP), which allow protons to leak back into the matrix and thus r...

  2. Uncoupling proteins, dietary fat and the metabolic syndrome

    OpenAIRE

    2006-01-01

    Abstract There has been intense interest in defining the functions of UCP2 and UCP3 during the nine years since the cloning of these UCP1 homologues. Current data suggest that both UCP2 and UCP3 proteins share some features with UCP1, such as the ability to reduce mitochondrial membrane potential, but they also have distinctly different physiological roles. Human genetic studies consistently demonstrate the effect of UCP2 alleles on type-2 diabetes. Less clear is whether UCP2 alleles influenc...

  3. Uncoupling proteins, dietary fat and the metabolic syndrome

    Directory of Open Access Journals (Sweden)

    Warden Craig H

    2006-09-01

    Full Text Available Abstract There has been intense interest in defining the functions of UCP2 and UCP3 during the nine years since the cloning of these UCP1 homologues. Current data suggest that both UCP2 and UCP3 proteins share some features with UCP1, such as the ability to reduce mitochondrial membrane potential, but they also have distinctly different physiological roles. Human genetic studies consistently demonstrate the effect of UCP2 alleles on type-2 diabetes. Less clear is whether UCP2 alleles influence body weight or body mass index (BMI with many studies showing a positive effect while others do not. There is strong evidence that both UCP2 and UCP3 protect against mitochondrial oxidative damage by reducing the production of reactive oxygen species. The evidence that UCP2 protein is a negative regulator of insulin secretion by pancreatic β-cells is also strong: increased UCP2 decreases glucose stimulated insulin secretion ultimately leading to β-cell dysfunction. UCP2 is also neuroprotective, reducing oxidative stress in neurons. UCP3 may also transport fatty acids out of mitochondria thereby protecting the mitochondria from fatty acid anions or peroxides. Current data suggest that UCP2 plays a role in the metabolic syndrome through down-regulation of insulin secretion and development of type-2 diabetes. However, UCP2 may protect against atherosclerosis through reduction of oxidative stress and both UCP2 and UCP3 may protect against obesity. Thus, these UCP1 homologues may both contribute to and protect from the markers of the metabolic syndrome.

  4. Cysteinyl-tRNA deacylation can be uncoupled from protein synthesis.

    Directory of Open Access Journals (Sweden)

    Alexandre David

    Full Text Available Aminoacyl-tRNA synthetases (ARSs are critical components of protein translation, providing ribosomes with aminoacyl-tRNAs. In return, ribosomes release uncharged tRNAs as ARS substrates. Here, we show that tRNA deacylation can be uncoupled from protein synthesis in an amino acid specific manner. While tRNAs coupled to radiolabeled Met, Leu Lys, or Ser are stable in cells following translation inhibition with arsenite, radiolabeled Cys is released from tRNA at a high rate. We discuss possible translation independent functions for tRNA(Cys.

  5. Genetic analysis on 3'-terminal flanking region of uncoupling protein 3 in different pig breeds

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The 3′-terminal flanking region of porcine uncoupling protein 3 (UCP3) was cloned, the sequence data revealed 15 nucleotide substitutions among Landrace and three Chinese native pig breeds named Neijiang, Minpig and Erhualian. The continuous 9 polymorphic sites were checked by PCR-RFLP, the results indicatedthat Erhualian had extraordinary gene frequency, presented most significant difference by χ2 test compared with Landrace, Largewhite, Neijiang and Minpig respectively, significant level compared with Meishan; and Meishan also had significant difference compared with Landrace and Minpig respectively. These results canbe concluded that Taihu pigs have special genetic characteristics among pig breeds.

  6. Biochemical characterization of the small hydrophobic protein of avian metapneumovirus

    Science.gov (United States)

    Avian metapneumovirus (aMPV) is a paramyxovirus that has three membrane-associate proteins: glycoprotein (G), fusion (F), and small hydrophobic (SH) proteins. Among them, the SH protein is a small type II integral membrane protein that is incorporated into virions and is only present in certain para...

  7. Mitochondrial efficiency and exercise economy following heat stress: a potential role of uncoupling protein 3.

    Science.gov (United States)

    Salgado, Roy M; Sheard, Ailish C; Vaughan, Roger A; Parker, Daryl L; Schneider, Suzanne M; Kenefick, Robert W; McCormick, James J; Gannon, Nicholas P; Van Dusseldorp, Trisha A; Kravitz, Len R; Mermier, Christine M

    2017-02-01

    Heat stress has been reported to reduce uncoupling proteins (UCP) expression, which in turn should improve mitochondrial efficiency. Such an improvement in efficiency may translate to the systemic level as greater exercise economy. However, neither the heat-induced improvement in mitochondrial efficiency (due to decrease in UCP), nor its potential to improve economy has been studied. Determine: (i) if heat stress in vitro lowers UCP3 thereby improving mitochondrial efficiency in C2C12 myocytes; (ii) whether heat acclimation (HA) in vivo improves exercise economy in trained individuals; and (iii) the potential improved economy during exercise at altitude. In vitro, myocytes were heat stressed for 24 h (40°C), followed by measurements of UCP3, mitochondrial uncoupling, and efficiency. In vivo, eight trained males completed: (i) pre-HA testing; (ii) 10 days of HA (40°C, 20% RH); and (iii) post-HA testing. Pre- and posttesting consisted of maximal exercise test and submaximal exercise at two intensities to assess exercise economy at 1600 m (Albuquerque, NM) and 4350 m. Heat-stressed myocytes displayed significantly reduced UCP3 mRNA expression and, mitochondrial uncoupling (77.1 ± 1.2%, P economy did not change at low and moderate exercise intensities. Our findings indicate that while heat-induced reduction in UCP3 improves mitochondrial efficiency in vitro, this is not translated to in vivo improvement of exercise economy at 1600 m or 4350 m.

  8. Regulation of Thermogenesis In Plants: The Interaction of Alternative Oxidase and Plant Uncoupling Mitochondrial Protein

    Institute of Scientific and Technical Information of China (English)

    Yan Zhu; Jianfei Lu; Jing Wang; Fu Chen; Feifan Leng; Hongyu Li

    2011-01-01

    Thermogenesis is a process of heat production in living organisms.It is rare in plants,but it does occur in some species of angiosperm.The heat iS generated via plant mitochondrial respiration.As possible Involvement in thermogenesis of mitochondrial factors,alternative oxidases(AOXs)and plant uncoupling mitochondrial proteins(PUMPs)have been well studied.AOXs and PUMPs are ubiquitously present in the inner membrane of plant mitochondria.They serve as two major energy dissipation systems that balance mitochondrial respiration and uncoupled phosphorylation by dissipating the H+ redox energy and proton electrochemical gradient(△μH+)as heat,respectively.AOXs and PUMPs exert similar physiological functions during homeothermic heat production in thermogenic plants.AOXs have five isoforms,while PUMPs have six.Both AOXS and PUMPS are encoded by small nuclear multigene families.Multiple isoforms are expressed in different tissues or organs.Extensive studies have been done in the area of thermogenesis in higher plants.In this review,we focus on the involvement and regulation of AOXs and PUMPs in thermogenesis.

  9. Mitochondrial Hormesis in Pancreatic β Cells: Does Uncoupling Protein 2 Play a Role?

    Directory of Open Access Journals (Sweden)

    Ning Li

    2012-01-01

    Full Text Available In pancreatic β cells, mitochondrial metabolism translates glucose sensing into signals regulating insulin secretion. Chronic exposure of β cells to excessive nutrients, namely, glucolipotoxicity, impairs β-cell function. This is associated with elevated ROS production from overstimulated mitochondria. Mitochondria are not only the major source of cellular ROS, they are also the primary target of ROS attacks. The mitochondrial uncoupling protein UCP2, even though its uncoupling properties are debated, has been associated with protective functions against ROS toxicity. Hormesis, an adaptive response to cellular stresses, might contribute to the protection against β-cell death, possibly limiting the development of type 2 diabetes. Mitochondrial hormesis, or mitohormesis, is a defense mechanism observed in ROS-induced stress-responses by mitochondria. In β cells, mitochondrial damages induced by sublethal exogenous H2O2 can induce secondary repair and defense mechanisms. In this context, UCP2 is a marker of mitohormesis, being upregulated following stress conditions. When overexpressed in nonstressed naïve cells, UCP2 confers resistance to oxidative stress. Whether treatment with mitohormetic inducers is sufficient to restore or ameliorate secretory function of β cells remains to be determined.

  10. Overexpression of uncoupling protein 3 in skeletal muscle protects against fat-induced insulin resistance

    Science.gov (United States)

    Choi, Cheol Soo; Fillmore, Jonathan J.; Kim, Jason K.; Liu, Zhen-Xiang; Kim, Sheene; Collier, Emily F.; Kulkarni, Ameya; Distefano, Alberto; Hwang, Yu-Jin; Kahn, Mario; Chen, Yan; Yu, Chunli; Moore, Irene K.; Reznick, Richard M.; Higashimori, Takamasa; Shulman, Gerald I.

    2007-01-01

    Insulin resistance is a major factor in the pathogenesis of type 2 diabetes and is strongly associated with obesity. Increased concentrations of intracellular fatty acid metabolites have been postulated to interfere with insulin signaling by activation of a serine kinase cascade involving PKCθ in skeletal muscle. Uncoupling protein 3 (UCP3) has been postulated to dissipate the mitochondrial proton gradient and cause metabolic inefficiency. We therefore hypothesized that overexpression of UCP3 in skeletal muscle might protect against fat-induced insulin resistance in muscle by conversion of intramyocellular fat into thermal energy. Wild-type mice fed a high-fat diet were markedly insulin resistant, a result of defects in insulin-stimulated glucose uptake in skeletal muscle and hepatic insulin resistance. Insulin resistance in these tissues was associated with reduced insulin-stimulated insulin receptor substrate 1– (IRS-1–) and IRS-2–associated PI3K activity in muscle and liver, respectively. In contrast, UCP3-overexpressing mice were completely protected against fat-induced defects in insulin signaling and action in these tissues. Furthermore, these changes were associated with a lower membrane-to-cytosolic ratio of diacylglycerol and reduced PKCθ activity in whole-body fat–matched UCP3 transgenic mice. These results suggest that increasing mitochondrial uncoupling in skeletal muscle may be an excellent therapeutic target for type 2 diabetes mellitus. PMID:17571165

  11. Prenatal cocaine exposure uncouples mGluR1 from Homer1 and Gq Proteins.

    Directory of Open Access Journals (Sweden)

    Kalindi Bakshi

    Full Text Available Cocaine exposure during gestation causes protracted neurobehavioral changes consistent with a compromised glutamatergic system. Although cocaine profoundly disrupts glutamatergic neurotransmission and in utero cocaine exposure negatively affects metabotropic glutamate receptor-type 1 (mGluR1 activity, the effect of prenatal cocaine exposure on mGluR1 signaling and the underlying mechanism responsible for the prenatal cocaine effect remain elusive. Using brains of the 21-day-old (P21 prenatal cocaine-exposed rats, we show that prenatal cocaine exposure uncouples mGluR1s from their associated synaptic anchoring protein, Homer1 and signal transducer, Gq/11 proteins leading to markedly reduced mGluR1-mediated phosphoinositide hydrolysis in frontal cortex (FCX and hippocampus. This prenatal cocaine-induced effect is the result of a sustained protein kinase C (PKC-mediated phosphorylation of mGluR1 on the serine residues. In support, phosphatase treatment of prenatal cocaine-exposed tissues restores whereas PKC-mediated phosphorylation of saline-treated synaptic membrane attenuates mGluR1 coupling to both Gq/11 and Homer1. Expression of mGluR1, Homer1 or Gα proteins was not altered by prenatal cocaine exposure. Collectively, these data indicate that prenatal cocaine exposure triggers PKC-mediated hyper-phosphorylation of the mGluR1 leading to uncoupling of mGluR1 from its signaling components. Hence, blockade of excessive PKC activation may alleviate abnormalities in mGluR1 signaling and restores mGluR1-regulated brain functions in prenatal cocaine-exposed brains.

  12. Prenatal cocaine exposure uncouples mGluR1 from Homer1 and Gq Proteins.

    Science.gov (United States)

    Bakshi, Kalindi; Parihar, Raminder; Goswami, Satindra K; Walsh, Melissa; Friedman, Eitan; Wang, Hoau-Yan

    2014-01-01

    Cocaine exposure during gestation causes protracted neurobehavioral changes consistent with a compromised glutamatergic system. Although cocaine profoundly disrupts glutamatergic neurotransmission and in utero cocaine exposure negatively affects metabotropic glutamate receptor-type 1 (mGluR1) activity, the effect of prenatal cocaine exposure on mGluR1 signaling and the underlying mechanism responsible for the prenatal cocaine effect remain elusive. Using brains of the 21-day-old (P21) prenatal cocaine-exposed rats, we show that prenatal cocaine exposure uncouples mGluR1s from their associated synaptic anchoring protein, Homer1 and signal transducer, Gq/11 proteins leading to markedly reduced mGluR1-mediated phosphoinositide hydrolysis in frontal cortex (FCX) and hippocampus. This prenatal cocaine-induced effect is the result of a sustained protein kinase C (PKC)-mediated phosphorylation of mGluR1 on the serine residues. In support, phosphatase treatment of prenatal cocaine-exposed tissues restores whereas PKC-mediated phosphorylation of saline-treated synaptic membrane attenuates mGluR1 coupling to both Gq/11 and Homer1. Expression of mGluR1, Homer1 or Gα proteins was not altered by prenatal cocaine exposure. Collectively, these data indicate that prenatal cocaine exposure triggers PKC-mediated hyper-phosphorylation of the mGluR1 leading to uncoupling of mGluR1 from its signaling components. Hence, blockade of excessive PKC activation may alleviate abnormalities in mGluR1 signaling and restores mGluR1-regulated brain functions in prenatal cocaine-exposed brains.

  13. Effect of uncoupler on assembly pathway for pigment-binding protein of bacterial photosynthetic membranes. [Rhodobacter capsulatus

    Energy Technology Data Exchange (ETDEWEB)

    Dierstein, R.; Drews, G.

    1986-10-01

    The uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) was used to investigate membrane protein assembly in the phototrophic bacterium Rhodobacter capsulatus. As found for Escherichia coli and mitochondrial proteins, assembly across the bacterial photosynthetic membranes was sensitive to CCCP. At uncoupler concentrations which were sufficient to block the export of the periplasmic cytochrome c/sub 2/ and an outer membrane protein, the integration of pigment-binding protein into the photosynthetic apparatus was abolished. The unassembled protein was detected on the inner surface of the intracytoplasmic membrane. After inactivation of CCCP, accumulated protein continued insertion into the membrane. The data suggest that after binding to the cytoplasmic face of the membrane (i), translocation of protein into a transmembrane orientation takes place (ii), which is a prerequisite for the formation of a functional pigment-protein complex (iii).

  14. An Arabidopsis mitochondrial uncoupling protein confers tolerance to drought and salt stress in transgenic tobacco plants.

    Directory of Open Access Journals (Sweden)

    Kevin Begcy

    Full Text Available BACKGROUND: Plants are challenged by a large number of environmental stresses that reduce productivity and even cause death. Both chloroplasts and mitochondria produce reactive oxygen species under normal conditions; however, stress causes an imbalance in these species that leads to deviations from normal cellular conditions and a variety of toxic effects. Mitochondria have uncoupling proteins (UCPs that uncouple electron transport from ATP synthesis. There is evidence that UCPs play a role in alleviating stress caused by reactive oxygen species overproduction. However, direct evidence that UCPs protect plants from abiotic stress is lacking. METHODOLOGY/PRINCIPAL FINDINGS: Tolerances to salt and water deficit were analyzed in transgenic tobacco plants that overexpress a UCP (AtUCP1 from Arabidopsis thaliana. Seeds of AtUCP1 transgenic lines germinated faster, and adult plants showed better responses to drought and salt stress than wild-type (WT plants. These phenotypes correlated with increased water retention and higher gas exchange parameters in transgenic plants that overexpress AtUCP1. WT plants exhibited increased respiration under stress, while transgenic plants were only slightly affected. Furthermore, the transgenic plants showed reduced accumulation of hydrogen peroxide in stressed leaves compared with WT plants. CONCLUSIONS/SIGNIFICANCE: Higher levels of AtUCP1 improved tolerance to multiple abiotic stresses, and this protection was correlated with lower oxidative stress. Our data support previous assumptions that UCPs reduce the imbalance of reactive oxygen species. Our data also suggest that UCPs may play a role in stomatal closure, which agrees with other evidence of a direct relationship between these proteins and photosynthesis. Manipulation of the UCP protein expression in mitochondria is a new avenue for crop improvement and may lead to crops with greater tolerance for challenging environmental conditions.

  15. Marked over expression of uncoupling protein-2 in beta cells exerts minor effects on mitochondrial metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Hals, Ingrid K., E-mail: ingrid.hals@ntnu.no [Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology (NTNU), Trondheim (Norway); Ogata, Hirotaka; Pettersen, Elin [Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology (NTNU), Trondheim (Norway); Ma, Zuheng; Bjoerklund, Anneli [Department of Molecular Medicine and Surgery, Karolinska Institute, Stockholm (Sweden); Skorpen, Frank [Department of Laboratory Medicine, NTNU, Trondheim (Norway); Egeberg, Kjartan Wollo [Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology (NTNU), Trondheim (Norway); Grill, Valdemar [Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology (NTNU), Trondheim (Norway); Department of Molecular Medicine and Surgery, Karolinska Institute, Stockholm (Sweden)

    2012-06-29

    Highlights: Black-Right-Pointing-Pointer The impact of UCP-2 over expression on mitochondrial function is controversial. Black-Right-Pointing-Pointer We tested mitochondrial functions at defined levels of overexpression. Black-Right-Pointing-Pointer We find minor increases of fatty acid oxidation and uncoupling. Black-Right-Pointing-Pointer Effects were seen only at high level (fourfold) of over expression. Black-Right-Pointing-Pointer Hence it is doubtful whether these effects are of importance in diabetes. -- Abstract: Evidence is conflicting as to the impact of elevated levels of uncoupling protein-2 (UCP-2) on insulin-producing beta cells. Here we investigated effects of a fourfold induction of UCP-2 protein primarily on mitochondrial parameters and tested for replication of positive findings at a lower level of induction. We transfected INS-1 cells to obtain a tet-on inducible cell line. A 48 h exposure to 1 {mu}g/ml of doxycycline (dox) induced UCP-2 fourfold (424 {+-} 113%, mean {+-} SEM) and 0.1 {mu}g/ml twofold (178 {+-} 29%, n = 3). Fourfold induced cells displayed normal viability (MTT, apoptosis), normal cellular insulin contents and, glucose-induced insulin secretion (+27 {+-} 11%) as well as D-[U-{sup 14}C]-glucose oxidation (+5 {+-} 9% at 11 mM glucose). Oxidation of [1-{sup 14}C]-oleate was increased from 4088 to 5797 fmol/{mu}g prot/2 h at 3.3 mM glucose, p < 0.03. Oxidation of L-[{sup 14}C(U)]-glutamine was unaffected. Induction of UCP-2 did not significantly affect measures of mitochondrial membrane potential (Rhodamine 123) or mitochondrial mass (Mitotracker Green) and did not affect ATP levels. Oligomycin-inhibited oxygen consumption (a measure of mitochondrial uncoupling) was marginally increased, the effect being significant in comparison with dox-only treated cells, p < 0.05. Oxygen radicals, assessed by dichlorofluorescin diacetate, were decreased by 30%, p < 0.025. Testing for the lower level of UCP-2 induction did not reproduce any of the

  16. Expression of Uncoupling Protein 2 in Breast Cancer Tissue and Drug-resistant Cells

    Institute of Scientific and Technical Information of China (English)

    Sun Yan; Yuan Yuan; Zhang Lili; Zhu Hong; Hu Sainan

    2013-01-01

    Objective:To explore the expression of uncoupling protein-2 (UCP2) in clinical breast cancer tissue and drug-resistant cells. Methods:The expression of UCP2 in breast cancer tissue and normal tissue adjacent to carcinoma as well as breast cancer cell MCF-7 and paclitaxel-resistant cell MX-1/T were respectively detected by immunohistochemistry and Western blot. Results:The expression of UCP2 in breast cancer tissue was signiifcantly higher than in normal tissue adjacent to carcinoma, and that in paclitaxel-resistant cell MX-1/T obviously higher than in breast cancer cell MCF-7. Conclusion:UCP2 is highly expressed in breast cancer tissue and drug-resistant cells.

  17. Cortical spreading depression produces a neuroprotective effect activating mitochondrial uncoupling protein-5

    Directory of Open Access Journals (Sweden)

    Viggiano E

    2016-07-01

    Full Text Available Emanuela Viggiano,1,2 Vincenzo Monda,1 Antonietta Messina,1 Fiorenzo Moscatelli,3 Anna Valenzano,3 Domenico Tafuri,4 Giuseppe Cibelli,3 Bruno De Luca,1 Giovanni Messina,1,3 Marcellino Monda1 1Department of Experimental Medicine, Section of Human Physiology and Unit of Dietetics and Sports Medicine, Second University of Naples, Naples, 2Department of Medicine, University of Padua, Padua, 3Department of Clinical and Experimental Medicine, University of Foggia, Foggia, 4Department of Motor Sciences and Wellness, University of Naples “Parthenope”, Naples, Italy Abstract: Depression of electrocorticogram propagating over the cortex surface results in cortical spreading depression (CSD, which is probably related to the pathophysiology of stroke, epilepsy, and migraine. However, preconditioning with CSD produces neuroprotection to subsequent ischemic episodes. Such effects require the expression or activation of several genes, including neuroprotective ones. Recently, it has been demonstrated that the expression of the uncoupling proteins (UCPs 2 and 5 is amplified during brain ischemia and their expression exerts a long-term effect upon neuron protection. To evaluate the neuroprotective consequence of CSD, the expression of UCP-5 in the brain cortex was measured following CSD induction. CSD was evoked in four samples of rats, which were sacrificed after 2 hours, 4 hours, 6 hours, and 24 hours. Western blot analyses were carried out to measure UCP-5 concentrations in the prefrontal cortices of both hemispheres, and immunohistochemistry was performed to determine the localization of UCP-5 in the brain cortex. The results showed a significant elevation in UCP-5 expression at 24 hours in all cortical strata. Moreover, UCP-5 was triggered by CSD, indicating that UCP-5 production can have a neuroprotective effect. Keywords: cortical spreading depression, neuroprotective effect, uncoupling protein-5

  18. Antigenic properties of avian hepatitis E virus capsid protein.

    Science.gov (United States)

    Zhao, Qin; Syed, Shahid Faraz; Zhou, En-Min

    2015-10-22

    Avian hepatitis E virus (HEV) is the main causative agent of big liver and spleen disease and hepatitis-splenomegaly syndrome in chickens, and is genetically and antigenically related to mammalian HEVs. HEV capsid protein contains immunodominant epitopes and induces a protective humoral immune response. A better understanding of the antigenic composition of this protein is critically important for the development of effective vaccine and sensitive and specific serological assays. To date, six linear antigenic domains (I-VI) have been characterized in avian HEV capsid protein and analyzed for their applications in the serological diagnosis and vaccine design. Domains I and V induce strong immune response in chickens and are common to avian, human, and swine HEVs, indicating that the shared epitopes hampering differential diagnosis of avian HEV infection. Domains III and IV are not immunodominant and elicit a weak immune response. Domain VI, located in the N-terminal region of the capsid protein, can also trigger an intense immune response, but the anti-domain VI antibodies are transient. The protection analysis showed that the truncated capsid protein containing the C-terminal 268 amino acid residues expressed by the bacterial system can provide protective immunity against avian HEV infection in chickens. However, the synthetic peptides incorporating the different linear antigenic domains (I-VI) and epitopes are non-protective. The antigenic composition of avian HEV capsid protein is altogether complex. To develop an effective vaccine and accurate serological diagnostic methods, more conformational antigenic domains or epitopes are to be characterized in detail.

  19. The role of uncoupling protein 3 regulating calcium ion uptake into mitochondria during sarcopenia

    Science.gov (United States)

    Nikawa, Takeshi; Choi, Inho; Haruna, Marie; Hirasaka, Katsuya; Maita Ohno, Ayako; Kondo Teshima, Shigetada

    Overloaded mitochondrial calcium concentration contributes to progression of mitochondrial dysfunction in aged muscle, leading to sarcopenia. Uncoupling protein 3 (UCP3) is primarily expressed in the inner membrane of skeletal muscle mitochondria. Recently, it has been reported that UCP3 is associated with calcium uptake into mitochondria. However, the mechanisms by which UCP3 regulates mitochondrial calcium uptake are not well understood. Here we report that UCP3 interacts with HS-1 associated protein X-1 (Hax-1), an anti-apoptotic protein that is localized in mitochondria, which is involved in cellular responses to calcium ion. The hydrophilic sequences within the loop 2, matrix-localized hydrophilic domain of mouse UCP3 are necessary for binding to Hax-1 of the C-terminal domain in adjacent to mitochondrial innermembrane. Interestingly, these proteins interaction occur the calcium-dependent manner. Indeed, overexpression of UCP3 significantly enhanced calcium uptake into mitochondria on Hax-1 endogenously expressing C2C12 myoblasts. In addition, Hax-1 knock-down enhanced calcium uptake into mitochondria on both UCP3 and Hax-1 endogenously expressing C2C12 myotubes, but not myoblasts. Finally, the dissociation of UCP3 and Hax-1 enhances calcium uptake into mitochondria in aged muscle. These studies identify a novel UCP3-Hax-1 complex regulates the influx of calcium ion into mitochondria in muscle. Thus, the efficacy of UCP3-Hax-1 in mitochondrial calcium regulation may provide a novel therapeutic approach against mitochondrial dysfunction-related disease containing sarcopenia.

  20. Metabolically inert perfluorinated fatty acids directly activate uncoupling protein 1 in brown-fat mitochondria.

    Science.gov (United States)

    Shabalina, Irina G; Kalinovich, Anastasia V; Cannon, Barbara; Nedergaard, Jan

    2016-05-01

    The metabolically inert perfluorinated fatty acids perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) can display fatty acid-like activity in biological systems. The uncoupling protein 1 (UCP1) in brown adipose tissue is physiologically (re)activated by fatty acids, including octanoate. This leads to bioenergetically uncoupled energy dissipation (heat production, thermogenesis). We have examined here the possibility that PFOA/PFOS can directly (re)activate UCP1 in isolated mouse brown-fat mitochondria. In wild-type brown-fat mitochondria, PFOS and PFOA overcame GDP-inhibited thermogenesis, leading to increased oxygen consumption and dissipated membrane potential. The absence of this effect in brown-fat mitochondria from UCP1-ablated mice indicated that it occurred through activation of UCP1. A competitive type of inhibition by increased GDP concentrations indicated interaction with the same mechanistic site as that utilized by fatty acids. No effect was observed in heart mitochondria, i.e., in mitochondria without UCP1. The stimulatory effect of PFOA/PFOS was not secondary to non-specific mitochondrial membrane permeabilization or to ROS production. Thus, metabolic effects of perfluorinated fatty acids could include direct brown adipose tissue (UCP1) activation. The possibility that this may lead to unwarranted extra heat production and thus extra utilization of food resources, leading to decreased fitness in mammalian wildlife, is discussed, as well as possible negative effects in humans. However, a possibility to utilize PFOA-/PFOS-like substances for activating UCP1 therapeutically in obesity-prone humans may also be envisaged.

  1. Control of mitochondrial pH by uncoupling protein 4 in astrocytes promotes neuronal survival.

    Science.gov (United States)

    Perreten Lambert, Hélène; Zenger, Manuel; Azarias, Guillaume; Chatton, Jean-Yves; Magistretti, Pierre J; Lengacher, Sylvain

    2014-11-07

    Brain activity is energetically costly and requires a steady and highly regulated flow of energy equivalents between neural cells. It is believed that a substantial share of cerebral glucose, the major source of energy of the brain, will preferentially be metabolized in astrocytes via aerobic glycolysis. The aim of this study was to evaluate whether uncoupling proteins (UCPs), located in the inner membrane of mitochondria, play a role in setting up the metabolic response pattern of astrocytes. UCPs are believed to mediate the transmembrane transfer of protons, resulting in the uncoupling of oxidative phosphorylation from ATP production. UCPs are therefore potentially important regulators of energy fluxes. The main UCP isoforms expressed in the brain are UCP2, UCP4, and UCP5. We examined in particular the role of UCP4 in neuron-astrocyte metabolic coupling and measured a range of functional metabolic parameters including mitochondrial electrical potential and pH, reactive oxygen species production, NAD/NADH ratio, ATP/ADP ratio, CO2 and lactate production, and oxygen consumption rate. In brief, we found that UCP4 regulates the intramitochondrial pH of astrocytes, which acidifies as a consequence of glutamate uptake, with the main consequence of reducing efficiency of mitochondrial ATP production. The diminished ATP production is effectively compensated by enhancement of glycolysis. This nonoxidative production of energy is not associated with deleterious H2O2 production. We show that astrocytes expressing more UCP4 produced more lactate, which is used as an energy source by neurons, and had the ability to enhance neuronal survival.

  2. Control of Mitochondrial pH by Uncoupling Protein 4 in Astrocytes Promotes Neuronal Survival*

    Science.gov (United States)

    Perreten Lambert, Hélène; Zenger, Manuel; Azarias, Guillaume; Chatton, Jean-Yves; Magistretti, Pierre J.; Lengacher, Sylvain

    2014-01-01

    Brain activity is energetically costly and requires a steady and highly regulated flow of energy equivalents between neural cells. It is believed that a substantial share of cerebral glucose, the major source of energy of the brain, will preferentially be metabolized in astrocytes via aerobic glycolysis. The aim of this study was to evaluate whether uncoupling proteins (UCPs), located in the inner membrane of mitochondria, play a role in setting up the metabolic response pattern of astrocytes. UCPs are believed to mediate the transmembrane transfer of protons, resulting in the uncoupling of oxidative phosphorylation from ATP production. UCPs are therefore potentially important regulators of energy fluxes. The main UCP isoforms expressed in the brain are UCP2, UCP4, and UCP5. We examined in particular the role of UCP4 in neuron-astrocyte metabolic coupling and measured a range of functional metabolic parameters including mitochondrial electrical potential and pH, reactive oxygen species production, NAD/NADH ratio, ATP/ADP ratio, CO2 and lactate production, and oxygen consumption rate. In brief, we found that UCP4 regulates the intramitochondrial pH of astrocytes, which acidifies as a consequence of glutamate uptake, with the main consequence of reducing efficiency of mitochondrial ATP production. The diminished ATP production is effectively compensated by enhancement of glycolysis. This nonoxidative production of energy is not associated with deleterious H2O2 production. We show that astrocytes expressing more UCP4 produced more lactate, which is used as an energy source by neurons, and had the ability to enhance neuronal survival. PMID:25237189

  3. Control of mitochondrial pH by uncoupling protein 4 in astrocytes promotes neuronal survival

    KAUST Repository

    Lambert, Hélène Perreten

    2014-09-18

    Brain activity is energetically costly and requires a steady and highly regulated flow of energy equivalents between neural cells. It is believed that a substantial share of cerebral glucose, the major source of energy of the brain, will preferentially be metabolized in astrocytes via aerobic glycolysis. The aim of this study was to evaluate whether uncoupling proteins (UCPs), located in the inner membrane of mitochondria, play a role in setting up the metabolic response pattern of astrocytes. UCPs are believed to mediate the transmembrane transfer of protons, resulting in the uncoupling of oxidative phosphorylation from ATP production. UCPs are therefore potentially important regulators of energy fluxes. The main UCP isoforms expressed in the brain are UCP2, UCP4, and UCP5. We examined in particular the role of UCP4 in neuron-astrocyte metabolic coupling and measured a range of functional metabolic parameters including mitochondrial electrical potential and pH, reactive oxygen species production, NAD/NADH ratio, ATP/ADP ratio, CO2 and lactate production, and oxygen consumption rate. In brief, we found that UCP4 regulates the intramitochondrial pH of astrocytes, which acidifies as a consequence of glutamate uptake, with the main consequence of reducing efficiency of mitochondrial ATP production. The diminished ATP production is effectively compensated by enhancement of glycolysis. This nonoxidative production of energy is not associated with deleterious H2O2 production. We show that astrocytes expressing more UCP4 produced more lactate, which is used as an energy source by neurons, and had the ability to enhance neuronal survival.

  4. The Role of Uncoupling Protein 2 During Myocardial Dysfunction in a Canine Model of Endotoxin Shock.

    Science.gov (United States)

    Wang, Xiaoting; Liu, Dawei; Chai, Wenzhao; Long, Yun; Su, Longxiang; Yang, Rongli

    2015-03-01

    To explore the role of uncoupling protein 2 (UCP2) during myocardial dysfunction in a canine model of endotoxin shock, 26 mongrel canines were randomly divided into the following four groups: A (control group; n = 6), B2 (shock after 2 h; n = 7), B4 (shock after 4 h; n = 7), and B6 (shock after 6 h; n = 6). Escherichia coli endotoxin was injected into the canines via the central vein, and hemodynamics were monitored. Energy metabolism, UCP2 mRNA and protein expression, and UCP2 localization were analyzed, and the correlation between energy metabolism changes, and UCP2 expression was determined. After the canine endotoxin shock model was successfully established, the expression of UCP2 mRNA and protein was found to increase, with later time points showing significant increases (P shock (P shock, and UCP2 may play an important role in this process. The negative correlation between UCP2 expression and energy metabolism requires further study, as the results might contribute to the treatment of sepsis with heart failure.

  5. Activity and functional interaction of alternative oxidase and uncoupling protein in mitochondria from tomato fruit

    Directory of Open Access Journals (Sweden)

    F.E. Sluse

    2000-03-01

    Full Text Available Cyanide-resistant alternative oxidase (AOX is not limited to plant mitochondria and is widespread among several types of protists. The uncoupling protein (UCP is much more widespread than previously believed, not only in tissues of higher animals but also in plants and in an amoeboid protozoan. The redox energy-dissipating pathway (AOX and the proton electrochemical gradient energy-dissipating pathway (UCP lead to the same final effect, i.e., a decrease in ATP synthesis and an increase in heat production. Studies with green tomato fruit mitochondria show that both proteins are present simultaneously in the membrane. This raises the question of a specific physiological role for each energy-dissipating system and of a possible functional connection between them (shared regulation. Linoleic acid, an abundant free fatty acid in plants which activates UCP, strongly inhibits cyanide-resistant respiration mediated by AOX. Moreover, studies of the evolution of AOX and UCP protein expression and of their activities during post-harvest ripening of tomato fruit show that AOX and plant UCP work sequentially: AOX activity decreases in early post-growing stages and UCP activity is decreased in late ripening stages. Electron partitioning between the alternative oxidase and the cytochrome pathway as well as H+ gradient partitioning between ATP synthase and UCP can be evaluated by the ADP/O method. This method facilitates description of the kinetics of energy-dissipating pathways and of ATP synthase when state 3 respiration is decreased by limitation of oxidizable substrate.

  6. Cell Death and Heart Failure in Obesity: Role of Uncoupling Proteins

    Science.gov (United States)

    Ruiz-Ramírez, Angélica; López-Acosta, Ocarol; Barrios-Maya, Miguel Angel

    2016-01-01

    Metabolic diseases such as obesity, metabolic syndrome, and type II diabetes are often characterized by increased reactive oxygen species (ROS) generation in mitochondrial respiratory complexes, associated with fat accumulation in cardiomyocytes, skeletal muscle, and hepatocytes. Several rodents studies showed that lipid accumulation in cardiac myocytes produces lipotoxicity that causes apoptosis and leads to heart failure, a dynamic pathological process. Meanwhile, several tissues including cardiac tissue develop an adaptive mechanism against oxidative stress and lipotoxicity by overexpressing uncoupling proteins (UCPs), specific mitochondrial membrane proteins. In heart from rodent and human with obesity, UCP2 and UCP3 may protect cardiomyocytes from death and from a state progressing to heart failure by downregulating programmed cell death. UCP activation may affect cytochrome c and proapoptotic protein release from mitochondria by reducing ROS generation and apoptotic cell death. Therefore the aim of this review is to discuss recent findings regarding the role that UCPs play in cardiomyocyte survival by protecting against ROS generation and maintaining bioenergetic metabolism homeostasis to promote heart protection. PMID:27642497

  7. Inhibition of Uncoupling Protein 2 Attenuates Cardiac Hypertrophy Induced by Transverse Aortic Constriction in Mice

    Directory of Open Access Journals (Sweden)

    Xiao-Bing Ji

    2015-07-01

    Full Text Available Background: Uncoupling protein 2 (UCP2 is critical in regulating energy metabolism. Due to the significant change in energy metabolism of myocardium upon pressure overload, we hypothesize that UCP2 could contribute to the etiology of cardiac hypertrophy. Methods: Adult male C57BL/6J mice were subjected to pressure overload by using transverse aortic constriction (TAC, and then received genipin (a UCP2 selective inhibitor; 25 mg/kg/d, ip or vehicle for three weeks prior to histologic assessment of myocardial hypertrophy. ATP concentration, ROS level, and myocardial apoptosis were also examined. A parallel set of experiments was also conducted in UCP2-/- mice. Results: TAC induced left ventricular hypertrophy, as reflected by increased ventricular weight/thickness and increased size of myocardial cell (vs. sham controls. ATP concentration was decreased; ROS level was increased. Apoptosis and fibrosis markers were increased. TAC increased mitochondrial UCP2 expression in the myocardium at both mRNA and protein levels. Genipin treatment attenuated cardiac hypertrophy and the histologic/biochemical changes described above. Hypertrophy and associated changes induced by TAC in UCP2-/- mice were much less pronounced than in WT mice. Conclusions: Blocking UCP2 expression attenuates cardiac hypertrophy induced by pressure overload.

  8. A novel amino acid and metabolomics signature in mice overexpressing muscle uncoupling protein 3.

    Science.gov (United States)

    Aguer, Céline; Piccolo, Brian D; Fiehn, Oliver; Adams, Sean H; Harper, Mary-Ellen

    2017-02-01

    Uncoupling protein 3 (UCP3) is highly selectively expressed in skeletal muscle and is known to lower mitochondrial reactive oxygen species and promote fatty acid oxidation; however, the global impact of UCP3 activity on skeletal muscle and whole-body metabolism have not been extensively studied. We utilized untargeted metabolomics to identify novel metabolites that distinguish mice overexpressing UCP3 in muscle, both at rest and after exercise regimens that challenged muscle metabolism, to potentially unmask subtle phenotypes. Male wild-type (WT) and muscle-specific UCP3-overexpressing transgenic (UCP3 Tg) C57BL/6J mice were compared with or without a 5 wk endurance training protocol at rest or after an acute exercise bout (EB). Skeletal muscle, liver, and plasma samples were analyzed by gas chromatography time-of-flight mass spectrometry. Discriminant metabolites were considered if within the top 99th percentile of variable importance measurements obtained from partial least-squares discriminant analysis models. A total of 80 metabolites accurately discriminated UCP3 Tg mice from WT when modeled within a specific exercise condition (i.e., untrained/rested, endurance trained/rested, untrained/EB, and endurance trained/EB). Results revealed that several amino acids and amino acid derivatives in skeletal muscle and plasma of UCP3 Tg mice (e.g., Asp, Glu, Lys, Tyr, Ser, Met) were significantly reduced after an EB; that metabolites associated with skeletal muscle glutathione/Met/Cys metabolism (2-hydroxybutanoic acid, oxoproline, Gly, and Glu) were altered in UCP3 Tg mice across all training and exercise conditions; and that muscle metabolite indices of dehydrogenase activity were increased in UCP3 Tg mice, suggestive of a shift in tissue NADH/NAD(+) ratio. The results indicate that mitochondrial UCP3 activity affects metabolism well beyond fatty acid oxidation, regulating biochemical pathways associated with amino acid metabolism and redox status. That select

  9. Expression of PPARα modifies fatty acid effects on insulin secretion in uncoupling protein-2 knockout mice

    Directory of Open Access Journals (Sweden)

    Chan Catherine B

    2007-03-01

    Full Text Available Abstract Aims/hypothesis In uncoupling protein-2 (UCP2 knockout (KO mice, protection of beta cells from fatty acid exposure is dependent upon transcriptional events mediated by peroxisome proliferator-activated receptor-α (PPARα. Methods PPARα expression was reduced in isolated islets from UCP2KO and wild-type (WT mice with siRNA for PPARα (siPPARα overnight. Some islets were also cultured with oleic or palmitic acid, then glucose stimulated insulin secretion (GSIS was measured. Expression of genes was examined by quantitative RT-PCR or immunoblotting. PPARα activation was assessed by oligonucleotide consensus sequence binding. Results siPPARα treatment reduced PPARα protein expression in KO and WT islets by >85%. In siPPARα-treated UCP2KO islets, PA but not OA treatment significantly decreased the insulin response to 16.5 mM glucose. In WT islets, siPPARα treatment did not modify GSIS in PA and OA exposed groups. In WT islets, PA treatment significantly increased UCP2 mRNA and protein expression. Both PA and OA treatment significantly increased PPARα expression in UCP2KO and WT islets but OA treatment augmented PPARα protein expression only in UCP2KO islets (p Conclusion These data show that the negative effect of saturated fatty acid on GSIS is mediated by PPARα/UCP2. Knockout of UCP2 protects beta-cells from PA exposure. However, in the absence of both UCP2 and PPARα even a short exposure (24 h to PA significantly impairs GSIS.

  10. Effect of Acupuncture on Uncoupling Protein 1 Gene Expression for Brown Adipose Tissue of Obese Rats

    Institute of Scientific and Technical Information of China (English)

    刘志诚; 孙凤岷; 赵东红; 张中成; 孙志; 吴海涛; 徐炳国; 朱苗花; 李朝军

    2003-01-01

    Objective: To explore the effects of acupuncture on the expression of uncoupling protein 1(UCP1) gene of brown adipose tissue (BAT) in obese rats. Methods: The expression of UCP1 gene of BAT was determined with RT-PCR technique. The changes of body weight, Lee′s index, body fat, and the expression of UCP1 gene of BAT in obese rats were observed before and after acupuncture. Resuits:The body weight, Lee′s index, body fat in obese rats were all markedly higher than those in normal rats,but the expression of UCP1 gene of BAT in obese rats was all lower than that in normal rats. There were negative correlation between the obesity index and the expression of UCP1 gene in BAT. After acupuncture the marked effect of weight loss was achieved while the expression of UCP1 gene of BAT obviously increased in obese rats. Conclusion: The abnormal reduction for expression of UCP1 gene of BAT might be an important cause for the obesity. To promote the expression of UCP1 in obese organism might be an important cellular and molecular mechanism in anti-obesity effect by acupuncture.

  11. Mitochondrial uncoupling protein 2 and pancreatic cancer: a new potential target therapy.

    Science.gov (United States)

    Donadelli, Massimo; Dando, Ilaria; Dalla Pozza, Elisa; Palmieri, Marta

    2015-03-21

    Overall 5-years survival of pancreatic cancer patients is nearly 5%, making this cancer type one of the most lethal neoplasia. Furthermore, the incidence rate of pancreatic cancer has a growing trend that determines a constant increase in the number of deceases caused by this pathology. The poor prognosis of pancreatic cancer is mainly caused by delayed diagnosis, early metastasis of tumor, and resistance to almost all tested cytotoxic drugs. In this respect, the identification of novel potential targets for new and efficient therapies should be strongly encouraged in order to improve the clinical management of pancreatic cancer. Some studies have shown that the mitochondrial uncoupling protein 2 (UCP2) is over-expressed in pancreatic cancer as compared to adjacent normal tissues. In addition, recent discoveries established a key role of UCP2 in protecting cancer cells from an excessive production of mitochondrial superoxide ions and in the promotion of cancer cell metabolic reprogramming, including aerobic glycolysis stimulation, promotion of cancer progression. These observations together with the demonstration that UCP2 repression can synergize with standard chemotherapy to inhibit pancreatic cancer cell growth provide the molecular rationale to consider UCP2 as a potential therapeutic target for pancreatic cancer. In this editorial, recent advances describing the relationship between cancer development and mitochondrial UCP2 activity are critically provided.

  12. Uncoupling protein 2 in the glial response to stress:implications for neuroprotection

    Institute of Scientific and Technical Information of China (English)

    Daniel T. Hass; Colin J. Barnstable

    2016-01-01

    Reactive oxygen species (ROS) are free radicals thought to mediate the neurotoxic effects of several neu-rodegenerative disorders. In the central nervous system, ROS can also trigger a phenotypic switch in both astrocytes and microglia that further aggravates neurodegeneration, termed reactive gliosis. Negative regulators of ROS, such as mitochondrial uncoupling protein 2 (UCP2) are neuroprotective factors that decrease neuron loss in models of stroke, epilepsy, and parkinsonism. However, it is unclear whether UCP2 acts purely to prevent ROS production, or also to prevent gliosis. In this review article, we discuss published evidence supporting the hypothesis that UCP2 is a neuroprotective factor both through its direct effects in decreasing mitochondrial ROS and through its effects in astrocytes and microglia. A major effect of UCP2 activation in glia is a change in the spectrum of secreted cytokines towards a more anti-inlfammatory spec-trum. There are multiple mechanisms that can control the level or activity of UCP2, including a variety of metabolites and microRNAs. Understanding these mechanisms will be key to exploitingthe protective effects of UCP2 in therapies for multiple neurodegenerative conditions.

  13. Low-level lasers affect uncoupling protein gene expression in skin and skeletal muscle tissues

    Science.gov (United States)

    Canuto, K. S.; Sergio, L. P. S.; Paoli, F.; Mencalha, A. L.; Fonseca, A. S.

    2016-03-01

    Wavelength, frequency, power, fluence, and emission mode determine the photophysical, photochemical, and photobiological responses of biological tissues to low-level lasers. Free radicals are involved in these responses acting as second messengers in intracellular signaling processes. Irradiated cells present defenses against these chemical species to avoid unwanted effects, such as uncoupling proteins (UCPs), which are part of protective mechanisms and minimize the effects of free radical generation in mitochondria. In this work UCP2 and UCP3 mRNA gene relative expression in the skin and skeletal muscle tissues of Wistar rats exposed to low-level red and infrared lasers was evaluated. Samples of the skin and skeletal muscle tissue of Wistar rats exposed to low-level red and infrared lasers were withdrawn for total RNA extraction, cDNA synthesis, and the evaluation of gene expression by quantitative polymerase chain reaction. UCP2 and UCP3 mRNA expression was differently altered in skin and skeletal muscle tissues exposed to lasers in a wavelength-dependent effect, with the UCP3 mRNA expression dose-dependent. Alteration on UCP gene expression could be part of the biostimulation effect and is necessary to make cells exposed to red and infrared low-level lasers more resistant or capable of adapting in damaged tissues or diseases.

  14. Rapamycin negatively impacts insulin signaling, glucose uptake and uncoupling protein-1 in brown adipocytes.

    Science.gov (United States)

    García-Casarrubios, Ester; de Moura, Carlos; Arroba, Ana I; Pescador, Nuria; Calderon-Dominguez, María; Garcia, Laura; Herrero, Laura; Serra, Dolors; Cadenas, Susana; Reis, Flavio; Carvalho, Eugenia; Obregon, Maria Jesus; Valverde, Ángela M

    2016-12-01

    New onset diabetes after transplantation (NODAT) is a metabolic disorder that affects 40% of patients on immunosuppressive agent (IA) treatment, such as rapamycin (also known as sirolimus). IAs negatively modulate insulin action in peripheral tissues including skeletal muscle, liver and white fat. However, the effects of IAs on insulin sensitivity and thermogenesis in brown adipose tissue (BAT) have not been investigated. We have analyzed the impact of rapamycin on insulin signaling, thermogenic gene-expression and mitochondrial respiration in BAT. Treatment of brown adipocytes with rapamycin for 16h significantly decreased insulin receptor substrate 1 (IRS1) protein expression and insulin-mediated protein kinase B (Akt) phosphorylation. Consequently, both insulin-induced glucose transporter 4 (GLUT4) translocation to the plasma membrane and glucose uptake were decreased. Early activation of the N-terminal Janus activated kinase (JNK) was also observed, thereby increasing IRS1 Ser 307 phosphorylation. These effects of rapamycin on insulin signaling in brown adipocytes were partly prevented by a JNK inhibitor. In vivo treatment of rats with rapamycin for three weeks abolished insulin-mediated Akt phosphorylation in BAT. Rapamycin also inhibited norepinephrine (NE)-induced lipolysis, the expression of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) and uncoupling protein (UCP)-1 in brown adipocytes. Importantly, basal mitochondrial respiration, proton leak and maximal respiratory capacity were significantly decreased in brown adipocytes treated with rapamycin. In conclusion, we demonstrate, for the first time the important role of brown adipocytes as target cells of rapamycin, suggesting that insulin resistance in BAT might play a major role in NODAT development.

  15. Exploring uncoupling proteins and antioxidant mechanisms under acute cold exposure in brains of fish.

    Directory of Open Access Journals (Sweden)

    Yung-Che Tseng

    Full Text Available Exposure to fluctuating temperatures accelerates the mitochondrial respiration and increases the formation of mitochondrial reactive oxygen species (ROS in ectothermic vertebrates including fish. To date, little is known on potential oxidative damage and on protective antioxidative defense mechanisms in the brain of fish under cold shock. In this study, the concentration of cellular protein carbonyls in brain was significantly increased by 38% within 1 h after cold exposure (from 28 °C to 18 °C of zebrafish (Danio rerio. In addition, the specific activity of superoxide dismutase (SOD and the mRNA level of catalase (CAT were increased after cold exposure by about 60% (6 h and by 60%-90% (1 and 24 h, respectively, while the specific glutathione content as well as the ratio of glutathione disulfide to glutathione remained constant and at a very low level. In addition, cold exposure increased the protein level of hypoxia-inducible factor (HIF by about 50% and the mRNA level of the glucose transporter zglut3 in brain by 50%-100%. To test for an involvement of uncoupling proteins (UCPs in the cold adaptation of zebrafish, five UCP members were annotated and identified (zucp1-5. With the exception of zucp1, the mRNA levels of the other four zucps were significantly increased after cold exposure. In addition, the mRNA levels of four of the fish homologs (zppar of the peroxisome proliferator-activated receptor (PPAR were increased after cold exposure. These data suggest that PPARs and UCPs are involved in the alterations observed in zebrafish brain after exposure to 18°C. The observed stimulation of the PPAR-UCP axis may help to prevent oxidative damage and to maintain metabolic balance and cellular homeostasis in the brains of ectothermic zebrafish upon cold exposure.

  16. Test systems to study the structure and function of uncoupling protein 1: a critical overview

    Directory of Open Access Journals (Sweden)

    Verena eHirschberg

    2011-11-01

    Full Text Available The discovery of active brown adipose tissue (BAT in healthy adult humans has renewed interest in the biology of this organ. BAT is capable of distributing nutrient energy in the form of heat allowing small mammals to efficiently defend their body temperature when acutely exposed to the cold. On the other hand BAT might be a target for the treatment of obesity and related diseases, as its pharmacological activation could allow release of excess energy stored in white adipose tissue depots. Energy dissipation in BAT depends on the activity of uncoupling protein 1 (UCP1, therefore a BAT-based obesity therapy requires a detailed understanding of structure and function of UCP1. Although UCP1 has been in the focus of research since its discovery, central questions concerning its mechanistic function and regulation are not yet resolved. They have been addressed in native mitochondria but also in several test systems, which are generally used to lower inter-experimental variability and to simplify analysis conditions. Different test systems have contributed to our current knowledge about UCP1 but of course all of them have certain limitations. We here provide an overview about research on UCP1 structure and function in test systems. So far, these have nearly exclusively been employed to study rodent and not human UCP1. Considering that the amino acid sequence of mouse and human UCP1 is only 79% identical, it will be essential to test whether the human version has a similarly high catalytic activity, allowing a relevant amount of energy dissipation in human BAT. Besides the issue of comparable mechanistic function a sufficiently high expression level of human UCP1 is a further prerequisite for anti-obesity therapeutic potential. Treatments which induce BAT hyperplasia and UCP1 expression in humans might therefore be equally important to discover as mere activators of the thermogenic process.

  17. Dietary Curcumin Ameliorates Aging-Related Cerebrovascular Dysfunction through the AMPK/Uncoupling Protein 2 Pathway

    Directory of Open Access Journals (Sweden)

    Yunfei Pu

    2013-11-01

    Full Text Available Background/Aims: Age-related cerebrovascular dysfunction contributes to stroke, cerebral amyloid angiopathy, cognitive decline and neurodegenerative diseases. One pathogenic mechanism underlying this effect is increased oxidative stress. Up-regulation of mitochondrial uncoupling protein 2 (UCP2 plays a crucial role in regulating reactive oxygen species (ROS production. Dietary patterns are widely recognized as contributors to cardiovascular and cerebrovascular disease. In this study, we tested the hypothesis that dietary curcumin, which has an antioxidant effect, can improve aging-related cerebrovascular dysfunction via UCP2 up-regulation. Methods: The 24-month-old male rodents used in this study, including male Sprague Dawley (SD rats and UCP2 knockout (UCP2-/- and matched wild type mice, were given dietary curcumin (0.2%. The young control rodents were 6-month-old. Rodent cerebral artery vasorelaxation was detected by wire myograph. The AMPK/UCP2 pathway and p-eNOS in cerebrovascular and endothelial cells were observed by immunoblotting. Results: Dietary curcumin administration for one month remarkably restored the impaired cerebrovascular endothelium-dependent vasorelaxation in aging SD rats. In cerebral arteries from aging SD rats and cultured endothelial cells, curcumin promoted eNOS and AMPK phosphorylation, up-regulated UCP2 and reduced ROS production. These effects of curcumin were abolished by either AMPK or UCP2 inhibition. Chronic dietary curcumin significantly reduced ROS production and improved cerebrovascular endothelium-dependent relaxation in aging wild type mice but not in aging UCP2-/- mice. Conclusions: Curcumin improves aging-related cerebrovascular dysfunction via the AMPK/UCP2 pathway.

  18. Endurance training blocks uncoupling protein 1 up-regulation in brown adipose tissue while increasing uncoupling protein 3 in the muscle tissue of rats fed with a high-sugar diet.

    OpenAIRE

    2012-01-01

    The mitochondrial uncoupling proteins (UCPs) of interscapular brown adipose tissue (iBAT) and of muscles play important roles in energy balance. For instance, the expression of UCP1 and UCP3 are modulated by free fatty acid gradients induced by high-sugar diets and acute exercise that is dependent on sympathetic stimulation. However, the effects of endurance training in animals fed with high-sugar diets are unknown. This study aims to evaluate the long-term effects of diet and exercise on UCP...

  19. Association of the sirtuin and mitochondrial uncoupling protein genes with carotid plaque.

    Directory of Open Access Journals (Sweden)

    Chuanhui Dong

    Full Text Available OBJECTIVE: Sirtuins (SIRTs and mitochondrial uncoupling proteins (UCPs have been implicated in cardiovascular diseases through the control of reactive oxygen species production. This study sought to investigate the association between genetic variants in the SIRT and UCP genes and carotid plaque. METHODS: In a group of 1018 stroke-free subjects from the Northern Manhattan Study with high-definition carotid ultrasonography and genotyping, we investigated the associations of 85 single nucleotide polymorphisms (SNPs in the 11 SIRT and UCP genes with the presence and number of carotid plaques, and evaluated interactions of SNPs with sex, smoking, diabetes and hypertension as well as interactions between SNPs significantly associated with carotid plaque. RESULTS: Overall, 60% of subjects had carotid plaques. After adjustment for demographic and vascular risk factors, T-carriers of the SIRT6 SNP rs107251 had an increased risk for carotid plaque (odds ratio, OR = 1.71, 95% CI = 1.23-2.37, Bonferroni-corrected p = 0.03 and for a number of plaques (rate ratio, RR = 1.31, 1.18-1.45, Bonferroni-corrected p = 1.4×10(-5, whereas T-carriers of the UCP5 SNP rs5977238 had an decreased risk for carotid plaque (OR = 0.49, 95% CI = 0.32-0.74, Bonferroni-corrected p = 0.02 and plaque number (RR = 0.64, 95% CI = 0.52-0.78, Bonferroni-corrected p = 4.9×10(-4. Some interactions with a nominal p≤0.01 were found between sex and SNPs in the UCP1 and UCP3 gene; between smoking, diabetes, hypertension and SNPs in UCP5 and SIRT5; and between SNPs in the UCP5 gene and the UCP1, SIRT1, SIRT3, SIRT5, and SIRT6 genes in association with plaque phenotypes. CONCLUSION: We observed significant associations between genetic variants in the SIRT6 and UCP5 genes and atherosclerotic plaque. We also found potential effect modifications by sex, smoking and vascular risk factors of the SIRT/UCP genes in the associations with atherosclerotic

  20. Methionine restriction decreases endogenous oxidative molecular damage and increases mitochondrial biogenesis and uncoupling protein 4 in rat brain.

    Science.gov (United States)

    Naudí, Alba; Caro, Pilar; Jové, Mariona; Gómez, José; Boada, Jordi; Ayala, Victoria; Portero-Otín, Manuel; Barja, Gustavo; Pamplona, Reinald

    2007-12-01

    Aging plays a central role in the occurrence of neurodegenerative diseases. Caloric restriction (CR) mitigates oxidative stress by decreasing the rate of generation of endogenous damage, a mechanism that can contribute to the slowing of the aging rate induced by this intervention. Various reports have recently linked methionine to aging, and methionine restriction (MetR) without energy restriction also increases life span. We have thus hypothesized that MetR can be responsible, at least in part, for the decrease in endogenous oxidative damage in CR. In this investigation we subjected male rats to exactly the same dietary protocol of MetR that is known to increase their life span. We have found that MetR: (1) decreases the mitochondrial complex I content and activity, as well as complex III content, while the complex II and IV, the mitochondrial flavoprotein apoptosis-inducing factor (AIF) and ATP content are unchanged; (2) increases the mitochondrial biogenesis factor PGC-1alpha; (3) increases the resistance of brain to metabolic and oxidative stress by increasing mitochondrial uncoupling protein 4 uncoupling protein 4 (UCP4); and (4) decreases mitochondrial oxidative DNA damage and all five different markers of protein oxidation measured and lowers membrane unsaturation in rat brain. No changes were detected for protein amino acid composition. These beneficial MetR-induced changes likely derived from metabolic reprogramming at the cellular and tissue level can play a key role in the protection against aging-associated neurodegenerative disorders.

  1. Expression of the mitochondrial uncoupling protein in brown adipocytes. Absence in brown preadipocytes and BFC-1 cells. Modulation by isoproterenol in adipocytes.

    Science.gov (United States)

    Forest, C; Doglio, A; Casteilla, L; Ricquier, D; Ailhaud, G

    1987-01-01

    The expression of the uncoupling protein has been compared in cells of BFC-1 clonal line established from mouse brown adipose tissue (BAT) and in preadipocytes, as well as in adipocytes from mouse BAT, both in primary culture. The results of immunoblots show that, after one week in culture, adipocytes have a reduced level of the 32 kD protein. This level can be raised 2-3.5-fold by a 24-h exposure to isoproterenol. Thus a direct modulation by a beta-agonist drug in the expression of the uncoupling protein is observed. Under the same conditions as well as under various other conditions, preadipocytes in primary culture and BFC-1 cells do not express the uncoupling protein. At the same time these cells are able both to differentiate into adipose cells, as demonstrated by the emergence of enzyme markers and triglyceride accumulation, and to respond to isoproterenol. Thus isoproterenol is not sufficient to trigger the expression of the uncoupling protein and behaves as a mere modulator once the cells have acquired the capacity to express it. Injection of undifferentiated BFC-1 cells into athymic mice bearing catecholamine-containing mini-osmotic pumps, or co-cultures of BFC-1 cells and pheochromocytoma PC-12 cells do not allow BFC-1 cells to express the uncoupling protein. Taken together, the results suggest that the formation of brown preadipocytes is critically linked during development to the release by sympathetic nerves of specific trophic factors acting locally.

  2. Spectrophotometric and Refractometric Determination of Total Protein in Avian Plasma

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    Rodica Căpriță

    2013-10-01

    Full Text Available The aim of this study was to compare the total protein values obtained in heparin plasma of chickens by a spectrophotometric technique (biuret method, and the values obtained on the same day in the same samples by refractometry. The results obtained by refractometry (average value 2.638±0.153g% were higher than those obtained by the spectrophotometric method (average value 2.441±0.181g%. There was a low correlation (r = 0.6709 between the total protein values, determined with both methods. Protein is the major determinant of plasma refractive index, but glucose contributes too. The refractometric method is not recommended in chickens for the determination of total protein, because avian blood glucose concentration averages about twice than in mammalian blood.

  3. Uncoupling protein-2 messenger ribonucleic acid expression during very-low-calorie diet in obese premenopausal women.

    Science.gov (United States)

    Barbe, P; Millet, L; Larrouy, D; Galitzky, J; Berlan, M; Louvet, J P; Langin, D

    1998-07-01

    Uncoupling protein-2 (UCP2) is a mitochondrial protein expressed in a wide range of human tissues. By uncoupling respiration from ATP synthesis, UCP2 might be involved in the control of energy expenditure. We have investigated UCP2 gene expression in human adipose tissue. In eight subjects, we found a positive correlation (r = 0.91, P < 0.002) between subcutaneous and visceral fat depots UCP2 messenger RNA (mRNA) levels, suggesting that UCP2 mRNA level in subcutaneous adipose tissue is a good index of UCP2 gene expression in whole body adipose tissues. The effect of a 25-day very-low-calorie diet un UCP2 mRNA level and resting metabolic rate was investigated in eight obese premenopausal women. There was no difference in UCP2 mRNA levels before and during the diet. After 25 days of hypocaloric diet, a positive correlation was found between adipose tissue UCP2 mRNA level and resting metabolic rate adjusted for lean body mass (r = 0.82, P < 0.01). These results show that very-low-calorie diet, unlike short-term fasting, is not associated with an induction in UCP2 mRNA expression, and that adipose tissue UCP2 mRNA levels may be related to variations in resting energy expenditure in humans.

  4. Acetoacetate reduces growth and ATP concentration in cancer cell lines which over-express uncoupling protein 2

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    Quadros Edward V

    2009-05-01

    Full Text Available Abstract Background Recent evidence suggests that several human cancers are capable of uncoupling of mitochondrial ATP generation in the presence of intact tricarboxylic acid (TCA enzymes. The goal of the current study was to test the hypothesis that ketone bodies can inhibit cell growth in aggressive cancers and that expression of uncoupling protein 2 is a contributing factor. The proposed mechanism involves inhibition of glycolytic ATP production via a Randle-like cycle while increased uncoupling renders cancers unable to produce compensatory ATP from respiration. Methods Seven aggressive human cancer cell lines, and three control fibroblast lines were grown in vitro in either 10 mM glucose medium (GM, or in glucose plus 10 mM acetoacetate [G+AcA]. The cells were assayed for cell growth, ATP production and expression of UCP2. Results There was a high correlation of cell growth with ATP concentration (r = 0.948 in a continuum across all cell lines. Controls demonstrated normal cell growth and ATP with the lowest density of mitochondrial UCP2 staining while all cancer lines demonstrated proportionally inhibited growth and ATP, and over-expression of UCP2 (p Conclusion Seven human cancer cell lines grown in glucose plus acetoacetate medium showed tightly coupled reduction of growth and ATP concentration. The findings were not observed in control fibroblasts. The observed over-expression of UCP2 in cancer lines, but not in controls, provides a plausible molecular mechanism by which acetoacetate spares normal cells but suppresses growth in cancer lines. The results bear on the hypothesized potential for ketogenic diets as therapeutic strategies.

  5. Genetic Variance in Uncoupling Protein 2 in Relation to Obesity, Type 2 Diabetes, and Related Metabolic Traits

    DEFF Research Database (Denmark)

    Dalgaard, Louise Torp

    2011-01-01

    and UCP3 genes with respect to obesity and diabetes. Of special interest was a promoter variant of UCP2 situated 866bp upstream of transcription initiation (-866G>A, rs659366). This variant changes promoter activity and has been associated with obesity and/or type 2 diabetes in several, although not all......Uncoupling proteins (UCPs) are mitochondrial proteins able to dissipate the proton gradient of the inner mitochondrial membrane when activated. This decreases ATP-generation through oxidation of fuels and may theoretically decrease energy expenditure leading to obesity. Evidence from Ucp......((-/-)) mice revealed a role of UCP2 in the pancreatic β-cell, because β-cells without UCP2 had increased glucose-stimulated insulin secretion. Thus, from being a candidate gene for obesity UCP2 became a valid candidate gene for type 2 diabetes mellitus. This prompted a series of studies of the human UCP2...

  6. Ubiquinol (QH(2)) functions as a negative regulator of purine nucleotide inhibition of Acanthamoeba castellanii mitochondrial uncoupling protein.

    Science.gov (United States)

    Woyda-Ploszczyca, Andrzej; Jarmuszkiewicz, Wieslawa

    2011-01-01

    We compared the influence of different adenine and guanine nucleotides on the free fatty acid-induced uncoupling protein (UCP) activity in non-phosphorylating Acanthamoeba castellanii mitochondria when the membranous ubiquinone (Q) redox state was varied. The purine nucleotides exhibit an inhibitory effect in the following descending order: GTP>ATP>GDP>ADP≫GMP>AMP. The efficiency of guanine and adenine nucleotides to inhibit UCP-sustained uncoupling in A. castellanii mitochondria depends on the Q redox state. Inhibition by purine nucleotides can be increased with decreasing Q reduction level (thereby ubiquinol, QH₂ concentration) even with nucleoside monophosphates that are very weak inhibitors at the initial respiration. On the other hand, the inhibition can be alleviated with increasing Q reduction level (thereby QH₂ concentration). The most important finding was that ubiquinol (QH₂) but not oxidised Q functions as a negative regulator of UCP inhibition by purine nucleotides. For a given concentration of QH₂, the linoleic acid-induced GTP-inhibited H(+) leak was the same for two types of A. castellanii mitochondria that differ in the endogenous Q content. When availability of the inhibitor (GTP) or the negative inhibition modulator (QH₂) was changed, a competitive influence on the UCP activity was observed. QH₂ decreases the affinity of UCP for GTP and, vice versa, GTP decreases the affinity of UCP for QH₂. These results describe the kinetic mechanism of regulation of UCP affinity for purine nucleotides by endogenous QH₂ in the mitochondria of a unicellular eukaryote.

  7. SET overexpression in HEK293 cells regulates mitochondrial uncoupling proteins levels within a mitochondrial fission/reduced autophagic flux scenario.

    Science.gov (United States)

    Almeida, Luciana O; Goto, Renata N; Neto, Marinaldo P C; Sousa, Lucas O; Curti, Carlos; Leopoldino, Andréia M

    2015-03-01

    We hypothesized that SET, a protein accumulated in some cancer types and Alzheimer disease, is involved in cell death through mitochondrial mechanisms. We addressed the mRNA and protein levels of the mitochondrial uncoupling proteins UCP1, UCP2 and UCP3 (S and L isoforms) by quantitative real-time PCR and immunofluorescence as well as other mitochondrial involvements, in HEK293 cells overexpressing the SET protein (HEK293/SET), either in the presence or absence of oxidative stress induced by the pro-oxidant t-butyl hydroperoxide (t-BHP). SET overexpression in HEK293 cells decreased UCP1 and increased UCP2 and UCP3 (S/L) mRNA and protein levels, whilst also preventing lipid peroxidation and decreasing the content of cellular ATP. SET overexpression also (i) decreased the area of mitochondria and increased the number of organelles and lysosomes, (ii) increased mitochondrial fission, as demonstrated by increased FIS1 mRNA and FIS-1 protein levels, an apparent accumulation of DRP-1 protein, and an increase in the VDAC protein level, and (iii) reduced autophagic flux, as demonstrated by a decrease in LC3B lipidation (LC3B-II) in the presence of chloroquine. Therefore, SET overexpression in HEK293 cells promotes mitochondrial fission and reduces autophagic flux in apparent association with up-regulation of UCP2 and UCP3; this implies a potential involvement in cellular processes that are deregulated such as in Alzheimer's disease and cancer.

  8. Identification of B-cell epitopes in the capsid protein of avian hepatitis E virus (avian HEV) that are common to human and swine HEVs or unique to avian HEV.

    Science.gov (United States)

    Guo, H; Zhou, E-M; Sun, Z F; Meng, X-J; Halbur, P G

    2006-01-01

    Avian hepatitis E virus (avian HEV) was recently discovered in chickens from the USA that had hepatitis-splenomegaly (HS) syndrome. The complete genomic sequence of avian HEV shares about 50 % nucleotide sequence identity with those of human and swine HEVs. The open reading frame 2 (ORF2) protein of avian HEV has been shown to cross-react with human and swine HEV ORF2 proteins, but the B-cell epitopes in the avian HEV ORF2 protein have not been identified. Nine synthetic peptides from the predicted four antigenic domains of the avian HEV ORF2 protein were synthesized and corresponding rabbit anti-peptide antisera were generated. Using recombinant ORF2 proteins, convalescent pig and chicken antisera, peptides and anti-peptide rabbit sera, at least one epitope at the C terminus of domain II (possibly between aa 477-492) that is unique to avian HEV, one epitope in domain I (aa 389-410) that is common to avian, human and swine HEVs, and one or more epitopes in domain IV (aa 583-600) that are shared between avian and human HEVs were identified. Despite the sequence difference in ORF2 proteins between avian and mammalian HEVs and similar ORF2 sequence between human and swine HEV ORF2 proteins, rabbit antiserum against peptide 6 (aa 389-399) recognized only human HEV ORF2 protein, suggesting complexity of the ORF2 antigenicity. The identification of these B-cell epitopes in avian HEV ORF2 protein may be useful for vaccine design and may lead to future development of immunoassays for differential diagnosis of avian, swine and human HEV infections.

  9. Features of uncoupling proteins%解偶联蛋白特征

    Institute of Scientific and Technical Information of China (English)

    闫润虎; 顾劲松; 于江; 顾威

    2008-01-01

    学术背景:机体产热部位主要在线粒体,产热量主要取决于线粒体上解偶联蛋白,即位于线粒体内膜介导H+内流的跨膜蛋白质形成的"质子漏"的程度.大概有25%ATP中的代谢能是由于质子遗漏现象而被转化成热量的形式散失的.通常测定机体产热量可知晓机体能量代谢的概况,进而可研究对肥胖产生的影响.目的:总结解偶联蛋白的作用机制、解偶联蛋白多态性与肥胖的关系及其表达的调节因素等,为肥胖发生机制及其防治工作的研究开辟新途径.检索策略:应用计算机检索Medline 1997-01/2007-07与解偶联蛋白相关的文章,检索词"Uncoupling proteins",限定文章语言种类为English;同时计算机检索1997-01/2007-07中国期刊全文数据库与解偶联蛋白相关的文章,检索词"解偶联蛋白",限定文章语言种类为中文.另外手工检索到相关文献20篇.对资料进行初审,纳入标准:①解偶联蛋白家族中各成员相关研究.②解偶联蛋白与肥胖方面的研究.③解偶联蛋白研究的临床、实验研究报道.排除标准:研究目的与本综述目的无关、重复研究的文献.阅读标题和摘要进行初筛,排除因研究目的与本研究无关者98篇,内容重复性的研究112篇,保留220篇中英文文献进一步分析.文献评价:220篇文献中动物实验和在体、离体、细胞学实验98篇,综述、述评、讲座类文献32篇,系统评价/Meta分析15篇,临床研究75篇.共34个研究满足全部纳入标准,予以纳入.资料综合:至今为止,在哺乳动物体内已识别出5种解偶联蛋白质.解偶联蛋白1在人体能量平衡中的作用不大.解偶联蛋白2和解偶联蛋白3与肥胖和2型糖尿病的发生可能有一定的相关性.解偶联蛋白4基因表达于大鼠脂肪组织中,并可能参与肥胖的发生发展.解偶联蛋白5则在脑与睾丸中含量高,可能与神经退行性病变有关,而与肥胖关系不大.摄食

  10. Exhaustive Training Increases Uncoupling Protein 2 Expression and Decreases Bcl-2/Bax Ratio in Rat Skeletal Muscle

    Directory of Open Access Journals (Sweden)

    W. Y. Liu

    2013-01-01

    Full Text Available This work investigates the effects of oxidative stress due to exhaustive training on uncoupling protein 2 (UCP2 and Bcl-2/Bax in rat skeletal muscles. A total of 18 Sprague-Dawley female rats were randomly divided into three groups: the control group (CON, the trained control group (TC, and the exhaustive trained group (ET. Malondialdehyde (MDA, superoxide dismutase (SOD, xanthine oxidase (XOD, ATPase, UCP2, and Bcl-2/Bax ratio in red gastrocnemius muscles were measured. Exhaustive training induced ROS increase in red gastrocnemius muscles, which led to a decrease in the cell antiapoptotic ability (Bcl-2/Bax ratio. An increase in UCP2 expression can reduce ROS production and affect mitochondrial energy production. Thus, oxidative stress plays a significant role in overtraining.

  11. Activation of Mitochondrial Uncoupling Protein 4 and ATP-Sensitive Potassium Channel Cumulatively Decreases Superoxide Production in Insect Mitochondria.

    Science.gov (United States)

    Slocińska, Malgorzata; Rosinski, Grzegorz; Jarmuszkiewicz, Wieslawa

    2016-01-01

    It has been evidenced that mitochondrial uncoupling protein 4 (UCP4) and ATP-regulated potassium channel (mKATP channel) of insect Gromphadorhina coqereliana mitochondria decrease superoxide anion production. We elucidated whether the two energy-dissipating systems work together on a modulation of superoxide level in cockroach mitochondria. Our data show that the simultaneous activation of UCP4 by palmitic acid and mKATP channel by pinacidil revealed a cumulative effect on weakening mitochondrial superoxide formation. The inhibition of UCP4 by GTP (and/or ATP) and mKATP channel by ATP elevated superoxide production. These results suggest a functional cooperation of both energy-dissipating systems in protection against oxidative stress in insects.

  12. Immunodominant epitopes mapped by synthetic peptides on the capsid protein of avian hepatitis E virus are non-protective.

    Science.gov (United States)

    Guo, Hailong; Zhou, E M; Sun, Z F; Meng, X J

    2008-03-01

    Avian hepatitis E virus (avian HEV) was recently discovered in chickens with hepatitis-splenomegaly syndrome in the United States. The open reading frame 2 (ORF2) protein of avian HEV has been shown to cross-react with human and swine HEV ORF2 proteins, and immunodominant antigenic epitopes on avian HEV ORF2 protein were identified in the predicted antigenic domains by synthetic peptides. However, whether these epitopes are protective against avian HEV infection has not been investigated. In this study, groups of chickens were immunized with keyhole limpet hemocyanin (KLH)-conjugated peptides and recombinant avian HEV ORF2 antigen followed by challenge with avian HEV virus to assess the protective capacity of these peptides containing the epitopes. While avian HEV ORF2 protein showed complete protection against infection, viremia and fecal virus shedding were found in all peptide-immunized chickens. Using purified IgY from normal, anti-peptide, and anti-avian HEV ORF2 chicken sera, an in-vitro neutralization and in-vivo monitoring assay was performed to further evaluate the neutralizing ability of anti-peptide IgY. Results showed that none of the anti-peptide IgY can neutralize avian HEV in vitro, as viremia, fecal virus shedding, and seroconversion appeared similarly in chickens inoculated with avian HEV mixed with anti-peptide IgY and chickens inoculated with avian HEV mixed with normal IgY. As expected, chickens inoculated with the avian HEV and anti-avian HEV ORF2 IgY mixture did not show detectable avian HEV infection. Taken together, the results of this study demonstrated that immunodominant epitopes on avian HEV ORF2 protein identified by synthetic peptides are non-protective, suggesting protective neutralizing epitope on avian HEV ORF2 may not be linear as is human HEV.

  13. p53 regulates expression of uncoupling protein 1 through binding and repression of PPARγ coactivator-1α

    DEFF Research Database (Denmark)

    Hallenborg, Philip; Fjære, Even; Liaset, Bjørn;

    2016-01-01

    1-dependent uncoupled respiration and increased oxidation of glucose and fatty acids in brown or brown-like, termed BRITE or beige, adipocytes in relation to energy balance and homeostasis has recently been highlighted. UCP1-dependent uncoupled respiration in classic interscapular brown adipose...

  14. SET overexpression in HEK293 cells regulates mitochondrial uncoupling proteins levels within a mitochondrial fission/reduced autophagic flux scenario

    Energy Technology Data Exchange (ETDEWEB)

    Almeida, Luciana O.; Goto, Renata N. [Department of Clinical Analyses, Toxicology and Food Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP (Brazil); Neto, Marinaldo P.C. [Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP (Brazil); Sousa, Lucas O. [Department of Clinical Analyses, Toxicology and Food Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP (Brazil); Curti, Carlos [Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP (Brazil); Leopoldino, Andréia M., E-mail: andreiaml@usp.br [Department of Clinical Analyses, Toxicology and Food Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP (Brazil)

    2015-03-06

    We hypothesized that SET, a protein accumulated in some cancer types and Alzheimer disease, is involved in cell death through mitochondrial mechanisms. We addressed the mRNA and protein levels of the mitochondrial uncoupling proteins UCP1, UCP2 and UCP3 (S and L isoforms) by quantitative real-time PCR and immunofluorescence as well as other mitochondrial involvements, in HEK293 cells overexpressing the SET protein (HEK293/SET), either in the presence or absence of oxidative stress induced by the pro-oxidant t-butyl hydroperoxide (t-BHP). SET overexpression in HEK293 cells decreased UCP1 and increased UCP2 and UCP3 (S/L) mRNA and protein levels, whilst also preventing lipid peroxidation and decreasing the content of cellular ATP. SET overexpression also (i) decreased the area of mitochondria and increased the number of organelles and lysosomes, (ii) increased mitochondrial fission, as demonstrated by increased FIS1 mRNA and FIS-1 protein levels, an apparent accumulation of DRP-1 protein, and an increase in the VDAC protein level, and (iii) reduced autophagic flux, as demonstrated by a decrease in LC3B lipidation (LC3B-II) in the presence of chloroquine. Therefore, SET overexpression in HEK293 cells promotes mitochondrial fission and reduces autophagic flux in apparent association with up-regulation of UCP2 and UCP3; this implies a potential involvement in cellular processes that are deregulated such as in Alzheimer's disease and cancer. - Highlights: • SET, UCPs and autophagy prevention are correlated. • SET action has mitochondrial involvement. • UCP2/3 may reduce ROS and prevent autophagy. • SET protects cell from ROS via UCP2/3.

  15. Relationship between expression of muscle-specific uncoupling protein 2 messenger RNA and genetic selection toward growth in channel catfish.

    Science.gov (United States)

    Kobayashi, Y; Peterson, B C; Waldbieser, G C

    2015-04-01

    This study tested the hypothesis that increased growth in channel catfish is associated with expression of the genes that code for uncoupling proteins (UCP) 2 and 3, members of the mitochondrial channel proteins involved in nutrient sensing and metabolism. The specific objective was to contrast the levels of UCP2 messenger RNA (mRNA) in fast vs slow growing catfish as well as in fed vs fasted catfish. Two distinct UCP2 transcripts were identified and named UCP2a and UCP2b, respectively. Nucleotide and amino acid sequence of catfish UCP2s were highly similar to UCP2 and other UCPs from other fish and mammals (>75%). Expression of UCP2a mRNA was detectable at very low levels in various metabolically active tissues, whereas the expression of UCP2b mRNA was readily detectable in the muscle and heart. In a 21-wk feeding study, fish that grew faster had a greater percent body fat at the end of the study (P muscle was increased (P growth and associated fat accumulation appears to be independent of muscle UCP2b mRNA expression and UCP2b-mediated mechanisms.

  16. Uncoupling Protein, UCP-4 May Be Involved in Neuronal Defects During Aging and Resistance to Pathogens in Caenorhabditis elegans

    Science.gov (United States)

    Cho, Injeong; Hwang, Gyu Jin; Cho, Jeong Hoon

    2016-01-01

    Uncoupling proteins (UCPs) are mitochondrial inner membrane proteins that function to dissipate proton motive force and mitochondrial membrane potential. One UCP has been identified in Caenorhabditis elegans (C. elegans), namely UCP-4. In this study, we examined its expression and localization using a GFP marker in C. elegans. ucp-4 was expressed throughout the body from early embryo to aged adult and UCP-4 was localized in the mitochondria. It is known that increased mitochondrial membrane protential leads to a reactive oxygen species (ROS) increase, which is associated with age-related diseases, including neurodegenerative diseases in humans. A ucp-4 mutant showed increased mitochondrial membrane protential in association with increased neuronal defects during aging, and the neurons of ucp-4 overexpressing animals showed decreased neuronal defects during aging. These results suggest that UCP-4 may be involved in neuroprotection during aging via relieving mitochondrial membrane protential. We also investigated the relationship between UCP-4 and innate immunity because increased ROS can affect innate immunity. ucp-4 mutant displayed increased resistance to the pathogen Staphylococcus aureus compared to wild type. The enhanced immunity in the ucp-4 mutant could be related to increased mitochondrial membrane protential, presumably followed by increased ROS. In summary, UCP-4 might have an important role in neuronal aging and innate immune responses through mediating mitochondrial membrane protential. PMID:27646689

  17. Uncoupling Protein, UCP-4 May Be Involved in Neuronal Defects During Aging and Resistance to Pathogens in Caenorhabditis elegans.

    Science.gov (United States)

    Cho, Injeong; Hwang, Gyu Jin; Cho, Jeong Hoon

    2016-09-01

    Uncoupling proteins (UCPs) are mitochondrial inner membrane proteins that function to dissipate proton motive force and mitochondrial membrane potential. One UCP has been identified in Caenorhabditis elegans (C. elegans), namely UCP-4. In this study, we examined its expression and localization using a GFP marker in C. elegans. ucp-4 was expressed throughout the body from early embryo to aged adult and UCP-4 was localized in the mitochondria. It is known that increased mitochondrial membrane protential leads to a reactive oxygen species (ROS) increase, which is associated with age-related diseases, including neurodegenerative diseases in humans. A ucp-4 mutant showed increased mitochondrial membrane protential in association with increased neuronal defects during aging, and the neurons of ucp-4 overexpressing animals showed decreased neuronal defects during aging. These results suggest that UCP-4 may be involved in neuroprotection during aging via relieving mitochondrial membrane protential. We also investigated the relationship between UCP-4 and innate immunity because increased ROS can affect innate immunity. ucp-4 mutant displayed increased resistance to the pathogen Staphylococcus aureus compared to wild type. The enhanced immunity in the ucp-4 mutant could be related to increased mitochondrial membrane protential, presumably followed by increased ROS. In summary, UCP-4 might have an important role in neuronal aging and innate immune responses through mediating mitochondrial membrane protential.

  18. Analysis of uncoupling protein 2-deficient mice upon anaesthesia and sedation revealed a role for UCP2 in locomotion.

    Directory of Open Access Journals (Sweden)

    Marie-Clotilde Alves-Guerra

    Full Text Available General anaesthesia is associated with hypothermia, oxidative stress, and immune depression. Uncoupling Protein (UCP2 is a member of the mitochondrial carrier family present in many organs including the spleen, the lung and the brain. A role of UCP2 in the activation of the inflammatory/immune cells, in the secretion of hormones, and in the excitability of neurons by regulating the production of reactive oxygen species has been discussed. Because of the side effects of anaesthesia listed above, we aimed to question the expression and the function of UCP2 during anaesthesia. Induction of anaesthesia with ketamine (20 mg/kg or isoflurane (3.6% and induction of sedation with the α2 adrenergic receptor agonist medetomidine (0.2 mg/kg stimulated infiltration of immune cells in the lung and increased UCP2 protein content in the lung, in both immune and non-immune cells. UCP2 content in the lung inversely correlated with body temperature decrease induced by medetomidine treatment. Challenge of the Ucp2(-/- mice with isoflurane and medetomidine revealed an earlier behavioral recovery phenotype. Transponder analysis of body temperature and activity showed no difference between Ucp2(-/- and control mice in basal conditions. However, upon an acute decrease of body temperature induced by medetomidine, Ucp2(-/- mice exhibited increased locomotion activity. Together, these results show that UCP2 is rapidly mobilized during anaesthesia and sedation in immune cells, and suggest a role of UCP2 in locomotion.

  19. The transcription factor Nrf2 promotes survival by enhancing the expression of uncoupling protein 3 under conditions of oxidative stress.

    Science.gov (United States)

    Anedda, Andrea; López-Bernardo, Elia; Acosta-Iborra, Bárbara; Saadeh Suleiman, M; Landázuri, Manuel O; Cadenas, Susana

    2013-08-01

    Uncoupling protein 3 (UCP3) is a member of the mitochondrial inner membrane carrier superfamily that modulates energy efficiency by catalyzing proton conductance and thus decreasing the production of superoxide anion. However, its role during oxidative stress and the underlying regulatory and molecular mechanisms remain poorly understood. We sought to investigate how UCP3 expression is regulated by oxidative stress and to evaluate the putative antioxidant role of this protein. H2O2 treatment increased UCP3 expression and the nuclear accumulation of the transcription factor Nrf2 in C2C12 and HL-1 cells. Nrf2 siRNA prevented H2O2-induced UCP3 expression, increasing oxidative stress and cell death. ChIP assays identified an antioxidant-response element (ARE) within the UCP3 promoter that bound Nrf2 after exposure to H2O2. Luciferase reporter experiments confirmed increased ARE activity in H2O2-treated HL-1 cells. Importantly, H2O2 increased the UCP3-mediated proton leak, suggesting a role for this protein in attenuating ROS-induced damage. Nrf2 nuclear accumulation and increased UCP3 protein were also detected in intact mouse heart subjected to a condition known to increase ROS generation. This is the first study to demonstrate that H2O2 augments UCP3 expression and it provides the first evidence of Nrf2 binding to the UCP3 promoter in response to oxidative challenge. These findings suggest that UCP3 functions as a member of the cellular antioxidant defense system that protects against oxidative stress in vivo. In conclusion, we have identified a novel regulatory process induced by an oxidative insult whereby the expression of the mitochondrial protein UCP3 is driven by the Nrf2 transcription factor, which decreases ROS production and prevents cell death.

  20. Identification of two neutralization epitopes on the capsid protein of avian hepatitis E virus.

    Science.gov (United States)

    Zhou, E-M; Guo, H; Huang, F F; Sun, Z F; Meng, X J

    2008-02-01

    Avian hepatitis E virus (avian HEV) is genetically and antigenically related to human HEV, the causative agent of hepatitis E. To identify the neutralizing epitopes on the capsid (ORF2) protein of avian HEV, four mAbs (7B2, 1E11, 10A2 and 5G10) against recombinant avian HEV ORF2 protein were generated. mAbs 7B2, 1E11 and 10A2 blocked each other for binding to avian HEV ORF2 protein in a competitive ELISA, whereas 5G10 did not block the other mAbs, suggesting that 7B2, 1E11 and 10A2 recognize the same or overlapping epitopes and 5G10 recognizes a different one. The epitopes recognized by 7B2, 1E11 and 10A2, and by 5G10 were mapped by Western blotting between aa 513 and 570, and between aa 476 and 513, respectively. mAbs 1E11, 10A2 and 5G10 were shown to bind to avian HEV particles in vitro, although only 5G10 reacted to viral antigens in transfected LMH cells. To assess the neutralizing activities of the mAbs, avian HEV was incubated in vitro with each mAb before inoculation into specific-pathogen-free chickens. Both viraemia and faecal virus shedding were delayed in chickens inoculated with the mixtures of avian HEV and 1E11, 10A2 or 5G10, suggesting that these three mAbs partially neutralize avian HEV.

  1. Different intracellular distribution of avian reovirus core protein sigmaA in cells of avian and mammalian origin

    Energy Technology Data Exchange (ETDEWEB)

    Vazquez-Iglesias, Lorena; Lostale-Seijo, Irene; Martinez-Costas, Jose [Departamento de Bioquimica y Biologia Molecular, Facultad de Farmacia, y Centro Singular de Investigacion en Quimica Biologica y Materiales Moleculares (CIQUS), Universidad de Santiago de Compostela, 15782-Santiago de Compostela (Spain); Benavente, Javier, E-mail: franciscojavier.benavente@usc.es [Departamento de Bioquimica y Biologia Molecular, Facultad de Farmacia, y Centro Singular de Investigacion en Quimica Biologica y Materiales Moleculares (CIQUS), Universidad de Santiago de Compostela, 15782-Santiago de Compostela (Spain)

    2012-10-25

    A comparative analysis of the intracellular distribution of avian reovirus (ARV) core protein sigmaA in cells of avian and mammalian origin revealed that, whereas the viral protein accumulates in the cytoplasm and nucleolus of avian cells, most sigmaA concentrates in the nucleoplasm of mammalian cells in tight association with the insoluble nuclear matrix fraction. Our results further showed that sigmaA becomes arrested in the nucleoplasm of mammalian cells via association with mammalian cell-specific factors and that this association prevents nucleolar targeting. Inhibition of RNA polymerase II activity, but not of RNA polymerase I activity, in infected mammalian cells induces nucleus-to-cytoplasm sigmaA translocation through a CRM1- and RanGTP-dependent mechanism, yet a heterokaryon assay suggests that sigmaA does not shuttle between the nucleus and cytoplasm. The scarcity of sigmaA in cytoplasmic viral factories of infected mammalian cells could be one of the factors contributing to limited ARV replication in mammalian cells.

  2. Genetic variants of uncoupling proteins-2 and -3 in relation to maximal oxygen uptake in different sports.

    Science.gov (United States)

    Holdys, Joanna; Gronek, Piotr; Kryściak, Jakub; Stanisławski, Daniel

    2013-01-01

    Uncoupling proteins 2 and 3 (UCP2 and UCP3) as mitochondrial electron transporters are involved in regulation of ATP production and energy dissipation as heat. Energy efficiency plays an important role in physical performance, especially in aerobic fitness. The aim of this study was to examine the association between maximal oxygen uptake and genetic variants of the UCP2 and UCP3 genes. The studies were carried out in a group of 154 men and 85 women, professional athletes representing various sports and fitness levels and students of the University of Physical Education in Poznań. Physiological and molecular procedures were used, i.e. direct measurement of maximum oxygen uptake (VO₂max) and analysis of an insertion/deletion (I/D) polymorphism in the 3'untranslated region of exon 8 of the UCP2 gene and a C>T substitution in exon 5 (Y210Y) of the UCP3 gene. No statistically significant associations were found, only certain trends. Insertion allele (I) of the I/D UCP2 and the T allele of the UCP3 gene were favourable in obtaining higher VO₂max level and might be considered as endurance-related alleles.

  3. Uncoupling protein 2 gene (UCP2) 45-bp I/D polymorphism is associated with adiposity among Malaysian women

    Indian Academy of Sciences (India)

    Yee-How Say; Zi-Lian Ban; Yogambigai Arumugam; Trishal Kaur; Mee-Lay Tan; Phee-Phee Chia; Sook-Ha Fan

    2014-12-01

    This study investigated the association of Uncoupling Protein 2 gene (UCP2) 45-bp I/D polymorphism with obesity and adiposity in 926 Malaysian subjects (416 males; 265 obese; 102/672/152 Malays/Chinese/Indians). The overall minor allele frequency (MAF) was 0.14, while MAFs according to Malay/Chinese/Indian were 0.17/0.12/0.21. The polymorphism was associated with ethnicity, obesity and overall adiposity (total body fat percentage, TBF), but not gender and central adiposity (waist–hip ratio, WHR). Gender- and ethnicity-stratified analysis revealed that within males, the polymorphism was not associated with ethnicity and anthropometric classes. However, within females, significantly more Indians, obese and those with high TBF carried I allele. Logistic regression analysis among females further showed the polymorphism was associated with obesity and overall adiposity; however, when adjusted for age and ethnicity, this association was abolished for obesity but remained significant for overall adiposity [Odds Ratio (OR) for ID genotype =2.02 (CI=1.18, 3.45; =0.01); I allele =1.81 (CI=1.15, 2.84; =0.01)]. Indeed, covariate analysis controlling for age and ethnicity also showed that those carrying ID genotype or I allele had significantly higher TBF than the rest. In conclusion, UCP2 45-bp I/D polymorphism is associated with overall adiposity among Malaysian women.

  4. Acute cold- and chronic heat-exposure upregulate hepatic leptin and muscle uncoupling protein (UCP) gene expression in broiler chickens.

    Science.gov (United States)

    Dridi, Sami; Temim, Soraya; Derouet, Michel; Tesseraud, Sophie; Taouis, Mohammed

    2008-08-01

    Emerging evidence showed that variations in environmental temperature affect both leptin and uncoupling protein (UCP) gene expression in mammals, whereas a little is known about such interactions in birds. Thus, we conducted the present study to investigate the influence of acute (2 hours) cold (4 degrees C) and chronic (10 days) heat (32 degrees C) exposure on hepatic leptin and muscle UCP gene expression in 5-wk-old broiler chickens. Both cold- and heat-exposure significantly (P < 0.05 to P < 0.001) upregulated hepatic leptin (by 35 and 46%, respectively) and muscle UCP mRNA levels (by 71 and 71%, respectively) compared to the thermoneutrality (22 degrees C). This result suggests that leptin and UCP may be involved in the thermoregulation response of chickens to extreme climate (cold and hot temperatures). The upregulation of hepatic leptin gene expression was accompanied by an increase in plasma leptin levels, indicating that leptin may be regulated at transcriptional level. The increase of leptin and UCP mRNA abundance, and leptinemia we report here were not related to plasma glucose or insulin levels. In conclusion, the exposure of broiler chickens to extreme ambient temperatures (cold and heat) increases hepatic leptin and muscle UCP gene expression.

  5. Effects of 2 G on adiposity, leptin, lipoprotein lipase, and uncoupling protein-1 in lean and obese Zucker rats

    Science.gov (United States)

    Warren, L. E.; Horwitz, B. A.; Hamilton, J. S.; Fuller, C. A.

    2001-01-01

    Male Zucker rats were exposed to 2 G for 8 wk to test the hypothesis that the leptin regulatory pathway contributes to recovery from effects of 2 G on feeding, growth, and nutrient partitioning. After initial hypophagia, body mass-independent food intake of the lean rats exposed to 2 G surpassed that of the lean rats maintained at 1 G, but food intake of the obese rats exposed to 2 G remained low. After 8 wk at 2 G, body mass and carcass fat were less in both genotypes. Leptin and percent fat were lower in lean rats exposed to 2 G vs. 1 G but did not differ in obese rats exposed to 2 G vs. 1 G. Although exposure to 2 G did not alter uncoupling protein-1 levels, it did elicit white fat pad-specific changes in lipoprotein lipase activity in obese but not lean rats. We conclude that 2 G affects both genotypes but that the lean Zucker rats recover their food intake and growth rate and retain "normal" lipoprotein lipase activity to a greater degree than do the obese rats, emphasizing the importance of a functional leptin regulatory pathway in this acclimation.

  6. A comparative approach to understanding tissue-specific expression of uncoupling protein 1 expression in adipose tissue

    Directory of Open Access Journals (Sweden)

    Andrew eShore

    2013-01-01

    Full Text Available The thermoregulatory function of brown adipose tissue (BAT is due to the tissue-specific expression of uncoupling protein 1 (UCP1 which is thought to have evolved in early mammals. We report that a CpG island close to the UCP1 transcription start site is highly conserved in all 29 vertebrates examined apart from the mouse and xenopus. Using methylation sensitive restriction digest and bisulphite mapping we show that the CpG island in both the bovine and human is largely un-methylated and is not related to differences in UCP1 expression between white and brown adipose tissue. Tissue-specific expression of UCP1 has been proposed to be regulated by a conserved 5’ distal enhancer which has been reported to be absent in marsupials. We demonstrate that the enhancer, is also absent in 5 eutherians as well as marsupials, monotremes, amphibians and fish, is present in pigs despite UCP1 having become a pseudogene, and that absence of the enhancer element does not relate to brown adipose tissue-specific UCP1 expression. We identify an additional putative 5’ regulatory unit which is conserved in 14 eutherian species but absent in other eutherians and vertebrates, but again unrelated to UCP1 expression. We conclude that despite clear evidence of conservation of regulatory elements in the UCP1 5’ untranslated region, this does not appear to be related to species or tissues-specific expression of UCP1.

  7. Endurance training blocks uncoupling protein 1 up-regulation in brown adipose tissue while increasing uncoupling protein 3 in the muscle tissue of rats fed with a high-sugar diet.

    Science.gov (United States)

    de Queiroz, Karina Barbosa; Rodovalho, Gisele Vieira; Guimarães, Juliana Bohnen; de Lima, Daniel Carvalho; Coimbra, Cândido Celso; Evangelista, Elísio Alberto; Guerra-Sá, Renata

    2012-09-01

    The mitochondrial uncoupling proteins (UCPs) of interscapular brown adipose tissue (iBAT) and of muscles play important roles in energy balance. For instance, the expression of UCP1 and UCP3 are modulated by free fatty acid gradients induced by high-sugar diets and acute exercise that is dependent on sympathetic stimulation. However, the effects of endurance training in animals fed with high-sugar diets are unknown. This study aims to evaluate the long-term effects of diet and exercise on UCP1 and UCP3 levels and energy balance efficiency. Rats fed with standard or high-sugar (HSD) diets were simultaneously subjected to running training over an 8-week period. After the training period, the rats were decapitated, and the iBAT and gastrocnemius muscle tissues were removed for evaluation of the β₃-receptor, Ucp1, and Ucp3 mRNA and protein expression, which were analyzed by quantitative reverse transcriptase polymerase chain reaction and Western blot, respectively. Groups fed with an HSD displayed a higher adiposity index and iBAT weight (P tissues. In association with an HSD, training restored the increasing β₃-receptor mRNA and greatly up-regulated the levels of Ucp3 mRNA. Therefore, training blocked the HSD-induced up-regulation of UCP1 expression in iBAT, whereas it up-regulated the expression of Ucp3 mRNA in muscle. These results suggest that training enhances the relationship between Ucp1/Ucp3 mRNA levels, which could result in higher energy efficiency, but not when HSD-induced elevated sympathetic activity is maintained.

  8. Genipin-induced inhibition of uncoupling protein-2 sensitizes drug-resistant cancer cells to cytotoxic agents.

    Directory of Open Access Journals (Sweden)

    Ryan J Mailloux

    Full Text Available Uncoupling protein-2 (UCP2 is known to suppress mitochondrial reactive oxygen species (ROS production and is employed by drug-resistant cancer cells to mitigate oxidative stress. Using the drug-sensitive HL-60 cells and the drug-resistant MX2 subline as model systems, we show that genipin, a UCP2 inhibitor, sensitizes drug-resistant cells to cytotoxic agents. Increased MX2 cell death was observed upon co-treatment with genipin and different doses of menadione, doxorubicin, and epirubicin. DCFH-DA fluorimetry revealed that the increase in MX2 cell death was accompanied by enhanced cellular ROS levels. The drug-induced increase in ROS was linked to genipin-mediated inhibition of mitochondrial proton leak. State 4 and resting cellular respiratory rates were higher in the MX2 cells in comparison to the HL-60 cells, and the increased respiration was readily suppressed by genipin in the MX2 cells. UCP2 accounted for a remarkable 37% of the resting cellular oxygen consumption indicating that the MX2 cells are functionally reliant on this protein. Higher amounts of UCP2 protein were detected in the MX2 versus the HL-60 mitochondria. The observed effects of genipin were absent in the HL-60 cells pointing to the selectivity of this natural product for drug-resistant cells. The specificity of genipin for UCP2 was confirmed using CHO cells stably expressing UCP2 in which genipin induced an ∼22% decrease in state 4 respiration. These effects were absent in empty vector CHO cells expressing no UCP2. Thus, the chemical inhibition of UCP2 with genipin sensitizes multidrug-resistant cancer cells to cytotoxic agents.

  9. A point mutation in Euglene gracilis chloroplast tRNA{sup Glu} uncouples protein and chlorophyll biosynthesis

    Energy Technology Data Exchange (ETDEWEB)

    Stange-Thomann, N.; Thomann, H.U.; Lloyd, A.J.; Soell, D. [Yale Univ., New Haven, CT (United States); Lyman, H. [Univ. of New York, Stony Brook, NY (United States)

    1994-08-16

    The universal precursor of tetrapyrrole pigments (e.g., chlorophylls and hemes) is 5-aminolevulinic acid (ALA), which in Euglena gracilis chloroplasts is derived via the two-step C{sub 5} pathway from glutamate charged to tRNA{sup Glu}. The first enzyme in this pathway, Glu-tRNA reductase (GluTR) catalyzes the reduction of glutamyl-tRNA{sup Glu} (Glu-tRNA) to glutamate 1-semialdehyde (GSA) with the release of the uncharged tRNA{sup Glu}. The second enzyme, GSA-2, 1-aminomutase, converts GSA to ALA. tRNA{sup Glu} is a specific cofactor for the NADPH-dependent reduction by GluTR, an enzyme that recognizes the tRNA in a sequence-specific manner. This RNA is the normal tRNA{sup Glu}, a dual-function molecule participating both in protein and in ALA and, hence, chlorophyll biosynthesis. A chlorophyll-deficient mutant of E. gracilis (Y{sub 9}ZNaIL) does not synthesize ALA from glutamate, although it contains GluTR and GSA-2,1-aminomutase activity. The tRNA{sup Glu} isolated from the mutant can still be acrylated with glutamate in vitro and in vitro. Furthermore, it supports chloroplast protein synthesis; however, it is a poor substrate for GluTR. Sequence analysis of the tRNA and of its gene revealed a C56 {yields} U mutation in the resulting gene product. C56 is therefore an important identity element for GluTR. Thus, a point mutation in the T loop of tRNA uncouples protein from chlorophyll biosynthesis.

  10. The Effect of Resveratrol and Quercetin Treatment on PPAR Mediated Uncoupling Protein (UCP-) 1, 2, and 3 Expression in Visceral White Adipose Tissue from Metabolic Syndrome Rats

    OpenAIRE

    2016-01-01

    Uncoupling proteins (UCPs) are members of the mitochondrial anion carrier superfamily involved in the control of body temperature and energy balance regulation. They are currently proposed as therapeutic targets for treating obesity and metabolic syndrome (MetS). We studied the gene expression regulation of UCP1, -2, and -3 in abdominal white adipose tissue (WAT) from control and MetS rats treated with two doses of a commercial mixture of resveratrol (RSV) and quercetin (QRC). We found that U...

  11. Protective Effect of Peroxisome Proliferator-Activated Receptor α Activation against Cardiac Ischemia-Reperfusion Injury Is Related to Upregulation of Uncoupling Protein-3

    OpenAIRE

    2016-01-01

    Activation of peroxisome proliferator-activated receptor α (PPARα) confers cardioprotection, while its mechanism remains elusive. We investigated the protective effect of PPARα activation against cardiac ischemia-reperfusion injury in terms of the expression of uncoupling protein (UCP). Myocardial infarct size and UCP expression were measured in rats treated with WY-14643 20 mg/kg, a PPARα ligand, or vehicle. WY-14643 increased UCP3 expression in vivo. Myocardial infarct size was decreased in...

  12. Alteration in cardiac uncoupling proteins and eNOS gene expression following high-intensity interval training in favor of increasing mechanical efficiency

    OpenAIRE

    Ali Asghar Fallahi; Shahnaz Shekarfroush; Mostafa Rahimi; Amirhossain Jalali; Ali Khoshbaten

    2016-01-01

    Objective(s): High-intensity interval training (HIIT) increases energy expenditure and mechanical energy efficiency. Although both uncoupling proteins (UCPs) and endothelial nitric oxide synthase (eNOS) affect the mechanical efficiency and antioxidant capacity, their effects are inverse. The aim of this study was to determine whether the alterations of cardiac UCP2, UCP3, and eNOS mRNA expression following HIIT are in favor of increased mechanical efficiency or decreased oxidative stress. Mat...

  13. Effects of trans-10, cis-12 conjugated linoleic acid on the expression of uncoupling proteins in hamsters fed an atherogenic diet

    OpenAIRE

    Ribot, Joan; Portillo Baquedano, María Puy; Picó, Catalina; Macarulla Arenaza, María Teresa; Palou, Andreu

    2007-01-01

    It is known that conjugated linoleic acid (CLA) feeding decreases body adiposity but the mechanisms involved are not clear. The aim of this study was to analyse whether alterations in uncoupling protein (UCP) expression in white and brown adipose tissues (WAT and BAT, respectively) and in skeletal muscle may be responsible for the effect of trans-10, cis-12 CLA on the size of body fat depots in hamsters. Animals were divided into three groups and fed an atherogenic diet with different amounts...

  14. Analysis of Uncoupling Protein 2-Deficient Mice upon Anaesthesia and Sedation Revealed a Role for UCP2 in Locomotion

    Science.gov (United States)

    Alves-Guerra, Marie-Clotilde; Aheng, Caroline; Pecqueur, Claire; Masscheleyn, Sandrine; Tharaux, Pierre Louis; Druilhe, Anne; Ricquier, Daniel; Challet, Etienne; Miroux, Bruno

    2012-01-01

    General anaesthesia is associated with hypothermia, oxidative stress, and immune depression. Uncoupling Protein (UCP2) is a member of the mitochondrial carrier family present in many organs including the spleen, the lung and the brain. A role of UCP2 in the activation of the inflammatory/immune cells, in the secretion of hormones, and in the excitability of neurons by regulating the production of reactive oxygen species has been discussed. Because of the side effects of anaesthesia listed above, we aimed to question the expression and the function of UCP2 during anaesthesia. Induction of anaesthesia with ketamine (20 mg/kg) or isoflurane (3.6%) and induction of sedation with the α2 adrenergic receptor agonist medetomidine (0.2 mg/kg) stimulated infiltration of immune cells in the lung and increased UCP2 protein content in the lung, in both immune and non-immune cells. UCP2 content in the lung inversely correlated with body temperature decrease induced by medetomidine treatment. Challenge of the Ucp2−/− mice with isoflurane and medetomidine revealed an earlier behavioral recovery phenotype. Transponder analysis of body temperature and activity showed no difference between Ucp2−/− and control mice in basal conditions. However, upon an acute decrease of body temperature induced by medetomidine, Ucp2−/− mice exhibited increased locomotion activity. Together, these results show that UCP2 is rapidly mobilized during anaesthesia and sedation in immune cells, and suggest a role of UCP2 in locomotion. PMID:22900002

  15. American Ginseng Stimulates Insulin Production and Prevents Apoptosis through Regulation of Uncoupling Protein-2 in Cultured β Cells

    Directory of Open Access Journals (Sweden)

    John Zeqi Luo

    2006-01-01

    Full Text Available American ginseng root displays the ability to achieve glucose homeostasis both experimentally and clinically but the unknown mechanism used by ginseng to achieve its therapeutic effects on diabetes limits its application. Disruption in the insulin secretion of pancreatic β cells is considered the major cause of diabetes. A mitochondrial protein, uncoupling protein-2 (UCP-2 has been found to play a critical role in insulin synthesis and β cell survival. Our preliminary studies found that the extracts of American ginseng inhibit UCP-2 expression which may contribute to the ability of ginseng protecting β cell death and improving insulin synthesis. Therefore, we hypothesized that ginseng extracts suppress UCP-2 in the mitochondria of pancreatic β cells, promoting insulin synthesis and anti-apoptosis (a programmed cell-death mechanism. To test the hypothesis, the serum-deprived quiescent β cells were cultured with or without interleukin-1β (IL-1β, (200 pg ml−1, a cytokine to induce β cell apoptosis and water extracts of American ginseng (25 μg per 5 μl administered to wells of 0.5 ml culture for 24 h. We evaluated effects of ginseng on UCP-2 expression, insulin production, anti-/pro-apoptotic factors Bcl-2/caspase-9 expression and cellular ATP levels. We found that ginseng suppresses UCP-2, down-regulates caspase-9 while increasing ATP and insulin production/secretion and up-regulates Bcl-2, reducing apoptosis. These findings suggest that stimulation of insulin production and prevention of β cell loss by American ginseng extracts can occur via the inhibition of mitochondrial UCP-2, resulting in increase in the ATP level and the anti-apoptotic factor Bcl-2, while down-regulation of pro-apoptotic factor caspase-9 occurs, lowering the occurrence of apoptosis, which support the hypothesis.

  16. Dimorphic expression of uncoupling protein-3 in golden hamster harderian gland: effects of castration and testosterone administration.

    Science.gov (United States)

    Santillo, Alessandra; Monteforte, Rossella; De Lange, Pieter; Lanni, Antonia; Farina, Paola; Baccari, Gabriella Chieffi

    2008-05-01

    Hamster (Mesocricetus auratus) harderian gland (HG) is a dimorphic orbital gland producing a copious lipid secretion. Two cell-types are present in hamster HG, type I in both sexes, type II only in males. In hamster HGs, we found a marked sexual dichotomy in the expression of uncoupling protein-3 (UCP3), a mitochondrial protein carrier, that probably exports fatty acid anions and fatty acid peroxides from the mitochondrial matrix. Following castration and/or testosterone treatment: (1) UCP3 levels correlated with the type II-cell percentage, not with testosterone levels, (2) in male HGs, UCP3 was comparable to female levels at 30 days post-castration (when the type II-cell percentage had fallen from 50 to 5%), although testosterone was already near zero at 15 days (when neither the type II-cell percentage nor the UCP3 level had fallen), and testosterone-replacement therapy prevented these changes. Testosterone-treated females possessed type II cells and a UCP3 level about twofold higher than in control females. Males displayed more intense UCP3 immunohistochemical positivity in type I HG cells than females. Hence, testosterone may indirectly control UCP3 expression by regulating the gland's morphological and lipid dimorphism. Straight-chain fatty acids [found in alkyl diacylglycerols (ADGs) in males] are oxidized predominantly in mitochondria, branched-chain fatty acids (abundant in ADGs in females) predominantly in peroxisomes, so we speculate that the higher UCP3 expression in males reflects greater fatty acid flux in HG mitochondria. This is supported by our finding that in female (not male) HGs, the peroxisome-rich fraction contained alpha-methylacyl-CoA racemase (AMACR), an enzyme important in the beta-oxidation of branched-chain fatty acids.

  17. Analysis of epitopes in the capsid protein of avian hepatitis E virus by using monoclonal antibodies.

    Science.gov (United States)

    Dong, Shiwei; Zhao, Qin; Lu, Mingzhe; Sun, Peiming; Qiu, Hongkai; Zhang, Lu; Lv, Junhua; Zhou, En-Min

    2011-02-01

    Avian hepatitis E virus (HEV) is related genetically and antigenically to human and swine HEVs and capsid protein of avian HEV shares approximately 48-49% amino acid sequence identities with those of human and swine HEVs. Six monoclonal antibodies (MAbs) were produced and used to locate different epitopes in the ORF2 region of aa 339-570 of avian HEV Chinese isolate. The results showed that five epitopes were located in the aa 339-414 region and one in the aa 510-515 region. Two epitopes located in aa 339-355 and aa 384-414 regions are the immunodominant epitopes on the surface of the avian HEV particles as demonstrated by immune capture of viral particles and immunohistochemical detection of the ORF2 antigens with two MAbs.

  18. 解偶联蛋白2的研究进展%Study progress on uncoupling protein 2

    Institute of Scientific and Technical Information of China (English)

    庄成君; 王力; 王冬梅; 田余祥

    2007-01-01

    解偶联蛋白(uncoupling protein,UCP)属于线粒体载体蛋白的亚类,包括UCP1~UCP5.UCPs定位在线粒体的内膜上.它们通过驱散质子梯度使呼吸链的氧化作用与ADP的磷酸化作用解偶联,减少ATP的合成.它们在氨基酸组成上有一定程度的同源性.UCPs借助于其信号序列,通过Tim10/Tim12/Tim22通道向线粒体内膜定向转运.UCPs的共同特征是每个单体具有6个跨膜域,二聚体发挥转运功能.但其表达的主要组织不同,主要的作用也不尽相同.特别是近年来的研究发现,UCP2参与能量代谢、子宫内膜退化、活性氧的产生、衰老等过程.并且与非乙醇性脂肪肝、动脉粥样硬化、局部缺血以及缺血再灌注损伤、抗肥胖及2型糖尿病等有着密切的关联性,故引起人们的广泛重视.

  19. Optimal response of key enzymes and uncoupling protein to cold in BAT depends on local T/sub 3/ generation

    Energy Technology Data Exchange (ETDEWEB)

    Bianco, A.C.; Silva, J.E.

    1987-09-01

    The authors have examined the activity of three lipogenic enzymes (malic enzyme (ME), glucose-6-phosphate dehydrogenase (G-6-PD), and acetyl coenzyme A (CoA) carboxylase), the activity of the mitochondrial FAD-dependent ..cap alpha..-glycerolphosphate dehydrogenase (..cap alpha..-GPD), and the mitochondrial concentration of uncoupling protein (UCP) in brown adipose tissue (BAT) of euthyroid and hypothyroid rats, both at room temperature and in response to acute cold stress. These enzymes and UCP are important for the thermogenic response of BAT in adaptation to cold. The basal level of the lipogenic enzymes was normal or slightly elevated in hypothyroid rats maintained at 23/sup 0/C, but the levels of ..cap alpha..-GPD and UCP were markedly reduced. Forty-eight hours at 4/sup 0/C resulted in an increase in the activity of G-6-PD, acetyl-CoA carboxylase, and ..cap alpha..-GPD and in the concentration of UCP both in euthyroid and hypothyroid animals, but the levels reached were invariably less in hypothyroid animals, indicating that thyroid hormone is necessary for a full metabolic response of BAT under maximal demands. Of all variables measured, the most affected was UCP followed by ..cap alpha..-GDP. Dose-response relationship analysis of the UCP response to T/sub 3/ indicated that the normalization of the response to cold requires saturation of the nuclear T/sub 3/ receptors. They concluded, therefore, that the activation of the BAT 5'-deiodinase induced by cold exposure is essential to provide the high levels of nuclear T/sub 3/ required for the full expression of BAT thermogenic potential.

  20. Peroxisome proliferator-activated receptor alpha induction of uncoupling protein 2 protects against acetaminophen-induced liver toxicity.

    Science.gov (United States)

    Patterson, Andrew D; Shah, Yatrik M; Matsubara, Tsutomu; Krausz, Kristopher W; Gonzalez, Frank J

    2012-07-01

    Acetaminophen (APAP) overdose causes acute liver failure in humans and rodents due in part to the destruction of mitochondria as a result of increased oxidative stress followed by hepatocellular necrosis. Activation of the peroxisome proliferator-activated receptor alpha (PPARα), a member of the nuclear receptor superfamily that controls the expression of genes encoding peroxisomal and mitochondrial fatty acid β-oxidation enzymes, with the experimental ligand Wy-14,643 or the clinically used fibrate drug fenofibrate, fully protects mice from APAP-induced hepatotoxicity. PPARα-humanized mice were also protected, whereas Ppara-null mice were not, thus indicating that the protection extends to human PPARα and is PPARα-dependent. This protection is due in part to induction of the PPARα target gene encoding mitochondrial uncoupling protein 2 (UCP2). Forced overexpression of UCP2 protected wildtype mice against APAP-induced hepatotoxicity in the absence of PPARα activation. Ucp2-null mice, however, were sensitive to APAP-induced hepatotoxicity despite activation of PPARα with Wy-14,643. Protection against hepatotoxicity by UCP2-induction through activation of PPARα is associated with decreased APAP-induced c-jun and c-fos expression, decreased phosphorylation of JNK and c-jun, lower mitochondrial H(2)O(2) levels, increased mitochondrial glutathione in liver, and decreased levels of circulating fatty acyl-carnitines. These studies indicate that the PPARα target gene UCP2 protects against elevated reactive oxygen species generated during drug-induced hepatotoxicity and suggest that induction of UCP2 may also be a general mechanism for protection of mitochondria during fatty acid β-oxidation.

  1. Glucocorticoids Suppress Mitochondrial Oxidant Production via Upregulation of Uncoupling Protein 2 in Hyperglycemic Endothelial Cells.

    Directory of Open Access Journals (Sweden)

    Domokos Gerö

    Full Text Available Diabetic complications are the leading cause of morbidity and mortality in diabetic patients. Elevated blood glucose contributes to the development of endothelial and vascular dysfunction, and, consequently, to diabetic micro- and macrovascular complications, because it increases the mitochondrial proton gradient and mitochondrial oxidant production. Therapeutic approaches designed to counteract glucose-induced mitochondrial reactive oxygen species (ROS production in the vasculature are expected to show efficacy against all diabetic complications, but direct pharmacological targeting (scavenging of mitochondrial oxidants remains challenging due to the high reactivity of some of these oxidant species. In a recent study, we have conducted a medium-throughput cell-based screening of a focused library of well-annotated pharmacologically active compounds and identified glucocorticoids as inhibitors of mitochondrial superoxide production in microvascular endothelial cells exposed to elevated extracellular glucose. The goal of the current study was to investigate the mechanism of glucocorticoids' action. Our findings show that glucocorticoids induce the expression of the mitochondrial UCP2 protein and decrease the mitochondrial potential. UCP2 silencing prevents the protective effect of the glucocorticoids on ROS production. UCP2 induction also increases the oxygen consumption and the "proton leak" in microvascular endothelial cells. Furthermore, glutamine supplementation augments the effect of glucocorticoids via further enhancing the expression of UCP2 at the translational level. We conclude that UCP2 induction represents a novel experimental therapeutic intervention in diabetic vascular complications. While direct repurposing of glucocorticoids may not be possible for the therapy of diabetic complications due to their significant side effects that develop during chronic administration, the UCP2 pathway may be therapeutically targetable by other

  2. Glucocorticoids Suppress Mitochondrial Oxidant Production via Upregulation of Uncoupling Protein 2 in Hyperglycemic Endothelial Cells

    Science.gov (United States)

    Szabo, Csaba

    2016-01-01

    Diabetic complications are the leading cause of morbidity and mortality in diabetic patients. Elevated blood glucose contributes to the development of endothelial and vascular dysfunction, and, consequently, to diabetic micro- and macrovascular complications, because it increases the mitochondrial proton gradient and mitochondrial oxidant production. Therapeutic approaches designed to counteract glucose-induced mitochondrial reactive oxygen species (ROS) production in the vasculature are expected to show efficacy against all diabetic complications, but direct pharmacological targeting (scavenging) of mitochondrial oxidants remains challenging due to the high reactivity of some of these oxidant species. In a recent study, we have conducted a medium-throughput cell-based screening of a focused library of well-annotated pharmacologically active compounds and identified glucocorticoids as inhibitors of mitochondrial superoxide production in microvascular endothelial cells exposed to elevated extracellular glucose. The goal of the current study was to investigate the mechanism of glucocorticoids' action. Our findings show that glucocorticoids induce the expression of the mitochondrial UCP2 protein and decrease the mitochondrial potential. UCP2 silencing prevents the protective effect of the glucocorticoids on ROS production. UCP2 induction also increases the oxygen consumption and the “proton leak” in microvascular endothelial cells. Furthermore, glutamine supplementation augments the effect of glucocorticoids via further enhancing the expression of UCP2 at the translational level. We conclude that UCP2 induction represents a novel experimental therapeutic intervention in diabetic vascular complications. While direct repurposing of glucocorticoids may not be possible for the therapy of diabetic complications due to their significant side effects that develop during chronic administration, the UCP2 pathway may be therapeutically targetable by other, glucocorticoid

  3. C-Terminal Amino Acids 471-507 of Avian Hepatitis E Virus Capsid Protein Are Crucial for Binding to Avian and Human Cells.

    Science.gov (United States)

    Zhang, Xinquan; Bilic, Ivana; Marek, Ana; Glösmann, Martin; Hess, Michael

    2016-01-01

    The infection of chickens with avian Hepatitis E virus (avian HEV) can be asymptomatic or induces clinical signs characterized by increased mortality and decreased egg production in adult birds. Due to the lack of an efficient cell culture system for avian HEV, the interaction between virus and host cells is still barely understood. In this study, four truncated avian HEV capsid proteins (ORF2-1 - ORF2-4) with an identical 338aa deletion at the N-terminus and gradual deletions from 0, 42, 99 and 136aa at the C-terminus, respectively, were expressed and used to map the possible binding site within avian HEV capsid protein. Results from the binding assay showed that three truncated capsid proteins attached to avian LMH cells, but did not penetrate into cells. However, the shortest construct, ORF2-4, lost the capability of binding to cells suggesting that the presence of amino acids 471 to 507 of the capsid protein is crucial for the attachment. The construct ORF2-3 (aa339-507) was used to study the potential binding of avian HEV capsid protein to human and other avian species. It could be demonstrated that ORF2-3 was capable of binding to QT-35 cells from Japanese quail and human HepG2 cells but failed to bind to P815 cells. Additionally, chicken serum raised against ORF2-3 successfully blocked the binding to LMH cells. Treatment with heparin sodium salt or sodium chlorate significantly reduced binding of ORF2-3 to LMH cells. However, heparinase II treatment of LMH cells had no effect on binding of the ORF2-3 construct, suggesting a possible distinct attachment mechanism of avian as compared to human HEV. For the first time, interactions between avian HEV capsid protein and host cells were investigated demonstrating that aa471 to 507 of the capsid protein are needed to facilitate interaction with different kind of cells from different species.

  4. Specificity and functional interaction of the polymerase complex proteins of human and avian metapneumoviruses

    NARCIS (Netherlands)

    M.T. de Graaf (Marieke); S. Herfst (Sander); E.J.A. Schrauwen (Eefje); Y. Choi (Ying); B.G. van den Hoogen (Bernadette); A.D.M.E. Osterhaus (Albert); R.A.M. Fouchier (Ron)

    2008-01-01

    textabstractHuman metapneumovirus (HMPV) and avian metapneumovirus (AMPV) have a similar genome organization and protein composition, but a different host range. AMPV subgroup C (AMPV-C) is more closely relaled to HMPV than other AMPVs. To investigate the specificity and functional interaction of th

  5. Use of G-protein-coupled and -uncoupled CCR5 receptors by CCR5 inhibitor-resistant and -sensitive human immunodeficiency virus type 1 variants.

    Science.gov (United States)

    Berro, Reem; Yasmeen, Anila; Abrol, Ravinder; Trzaskowski, Bartosz; Abi-Habib, Sarya; Grunbeck, Amy; Lascano, Danny; Goddard, William A; Klasse, Per Johan; Sakmar, Thomas P; Moore, John P

    2013-06-01

    Small-molecule CCR5 inhibitors such as vicriviroc (VVC) and maraviroc (MVC) are allosteric modulators that impair HIV-1 entry by stabilizing a CCR5 conformation that the virus recognizes inefficiently. Viruses resistant to these compounds are able to bind the inhibitor-CCR5 complex while also interacting with the free coreceptor. CCR5 also interacts intracellularly with G proteins, as part of its signal transduction functions, and this process alters its conformation. Here we investigated whether the action of VVC against inhibitor-sensitive and -resistant viruses is affected by whether or not CCR5 is coupled to G proteins such as Gαi. Treating CD4(+) T cells with pertussis toxin to uncouple the Gαi subunit from CCR5 increased the potency of VVC against the sensitive viruses and revealed that VVC-resistant viruses use the inhibitor-bound form of Gαi-coupled CCR5 more efficiently than they use uncoupled CCR5. Supportive evidence was obtained by expressing a signaling-deficient CCR5 mutant with an impaired ability to bind to G proteins, as well as two constitutively active mutants that activate G proteins in the absence of external stimuli. The implication of these various studies is that the association of intracellular domains of CCR5 with the signaling machinery affects the conformation of the external and transmembrane domains and how they interact with small-molecule inhibitors of HIV-1 entry.

  6. Uncoupling of bait-protein expression from the prey protein environment adds versatility for cell and tissue interaction proteomics and reveals a complex of CARP-1 and the PKA Cbeta1 subunit.

    Science.gov (United States)

    Erlbruch, Andrea; Hung, Chien-Wen; Seidler, Joerg; Borrmann, Katrin; Gesellchen, Frank; König, Norbert; Kübler, Dieter; Herberg, Friedrich W; Lehmann, Wolf D; Bossemeyer, Dirk

    2010-08-01

    An expression-uncoupled tandem affinity purification assay is introduced which differs from the standard TAP assay by uncoupling the expression of the TAP-bait protein from the target cells. Here, the TAP-tagged bait protein is expressed in Escherichia coli and purified. The two concatenated purification steps of the classical TAP are performed after addition of the purified bait to brain tissue homogenates, cell and nuclear extracts. Without prior genetic manipulation of the target, upscaling, free choice of cell compartments and avoidance of expression triggered heat shock responses could be achieved in one go. By the strategy of separating bait expression from the prey protein environment numerous established, mostly tissue-specific binding partners of the protein kinase A catalytic subunit Cbeta1 were identified, including interactions in binary, ternary and quaternary complexes. In addition, the previously unknown small molecule inhibitor-dependent interaction of Cbeta1 with the cell cycle and apoptosis regulatory protein-1 was verified. The uncoupled tandem affinity purification procedure presented here expands the application range of the in vivo TAP assay and may serve as a simple strategy for identifying cell- and tissue-specific protein complexes.

  7. Single nucleotide polymorphisms linked to mitochondrial uncoupling protein genes UCP2 and UCP3 affect mitochondrial metabolism and healthy aging in female nonagenarians.

    Science.gov (United States)

    Kim, Sangkyu; Myers, Leann; Ravussin, Eric; Cherry, Katie E; Jazwinski, S Michal

    2016-08-01

    Energy expenditure decreases with age, but in the oldest-old, energy demand for maintenance of body functions increases with declining health. Uncoupling proteins have profound impact on mitochondrial metabolic processes; therefore, we focused attention on mitochondrial uncoupling protein genes. Alongside resting metabolic rate (RMR), two SNPs in the promoter region of UCP2 were associated with healthy aging. These SNPs mark potential binding sites for several transcription factors; thus, they may affect expression of the gene. A third SNP in the 3'-UTR of UCP3 interacted with RMR. This UCP3 SNP is known to impact UCP3 expression in tissue culture cells, and it has been associated with body weight and mitochondrial energy metabolism. The significant main effects of the UCP2 SNPs and the interaction effect of the UCP3 SNP were also observed after controlling for fat-free mass (FFM) and physical-activity related energy consumption. The association of UCP2/3 with healthy aging was not found in males. Thus, our study provides evidence that the genetic risk factors for healthy aging differ in males and females, as expected from the differences in the phenotypes associated with healthy aging between the two sexes. It also has implications for how mitochondrial function changes during aging.

  8. The uncoupling protein 1 gene (UCP1 is disrupted in the pig lineage: a genetic explanation for poor thermoregulation in piglets.

    Directory of Open Access Journals (Sweden)

    Frida Berg

    2006-08-01

    Full Text Available Piglets appear to lack brown adipose tissue, a specific type of fat that is essential for nonshivering thermogenesis in mammals, and they rely on shivering as the main mechanism for thermoregulation. Here we provide a genetic explanation for the poor thermoregulation in pigs as we demonstrate that the gene for uncoupling protein 1 (UCP1 was disrupted in the pig lineage. UCP1 is exclusively expressed in brown adipose tissue and plays a crucial role for thermogenesis by uncoupling oxidative phosphorylation. We used long-range PCR and genome walking to determine the complete genome sequence of pig UCP1. An alignment with human UCP1 revealed that exons 3 to 5 were eliminated by a deletion in the pig sequence. The presence of this deletion was confirmed in all tested domestic pigs, as well as in European wild boars, Bornean bearded pigs, wart hogs, and red river hogs. Three additional disrupting mutations were detected in the remaining exons. Furthermore, the rate of nonsynonymous substitutions was clearly elevated in the pig sequence compared with the corresponding sequences in humans, cattle, and mice, and we used this increased rate to estimate that UCP1 was disrupted about 20 million years ago.

  9. Molecular characterization and expression of a novel homolog of uncoupling protein 5 (UCP5) from the eastern oyster Crassostrea virginica (Bivalvia: Ostreidae).

    Science.gov (United States)

    Kern, Britt; Ivanina, Anna V; Piontkivska, Helen; Sokolov, Eugene P; Sokolova, Inna M

    2009-06-01

    Uncoupling proteins (UCPs) belong to the mitochondrial anion carrier gene family which has been implicated in diverse physiological functions ranging from thermoregulation to antioxidant defense. In mammals, the UCP family is well characterized and contains five members (UCP1-5). In contrast, invertebrate homologues of uncoupling proteins are much less studied both from the viewpoints of structure and function. In this study we report nucleotide and predicted protein structure of an important member of UCP family, UCP5 from eastern oysters Crassostrea virginica. UCP5 from oysters appears to be a close homolog of the mammalian brain mitochondrial carrier protein (BMCP1, or UCP5) and is the first full-length UCP described from a Lophotrochozoan invertebrate. Evolutionary analysis of UCP sequences indicates at least three monophyletic UCP branches (UCP1-3, UCP4 and UCP5) that have diverged early in the evolution, prior to the divergence of vertebrates and invertebrates. In oysters, two forms of UCP5 transcript are found (UCP5S and UCP5L) that differ by 152 bp in length due to the presence of an intron in UCP5L. UCP5 was expressed in all studied oyster tissues, unlike mammals, where UCP5 is predominantly expressed in brains and male gonads. Hypoxia-reoxygenation stress, sublethal Cd exposure (50 ?g L(?1) Cd for 56 days) and acclimation to different temperatures (12 and 20 °C) had no significant effect on UCP5 mRNA expression in oysters indicative of its relative unimportance in antioxidant defense and temperature adaptation of oyster mitochondria. These data suggest that despite the relatively high degree of evolutionary conservation of the UCP5 amino acid sequence, its functional significance in mitochondria changed in the course of evolution of mollusks and vertebrates.

  10. Common genetic variants of the mitochondrial trafficking system and mitochondrial uncoupling proteins affect the development of two slowly developing demyelinating disorders, leukoaraiosis and multiple sclerosis.

    Science.gov (United States)

    Szolnoki, Z

    2010-01-01

    As the central energy source, the mitochondria are of great importance in the maintenance of the glia cells of the brain. It is presumed that mitochondrial energy production is affected not only by well-characterized genetic mutations of the mitochondria, which are associated with severe malfunctions and resultant acute glia and neuronal cell death, but also by a number of other unfavorable genetic variants. The genetic variants of the kinesin motor proteins and mitochondrial uncoupling proteins (UCPs) are believed to influence the mitochondrial energy production in different distress states of the glia cells. The kinesin motor proteins carry the mitochondria from the central parts to the peripheral parts of the glia cells, where myelin protein synthesis takes place. The UCPs are essential for regulation of the mitochondrial membrane potential under different physiological conditions, thereby finally attuning mitochondrial energy production in environmental states such as cold exposure, fasting or chronic mild hypoxia. While the capacity of the kinesin motor proteins can affect the number of mitochondria in the peripheral parts of the glia cells, the functional features of the UCPs can affect the degree of energy production of the mitochondria by influencing the mitochondrial membrane potential. The different genetic variants may display different activities, and some may result in a slowly developing energy shortage in the glia cells. In this context, this article discusses the roles of genetic variants of the kinesin motor proteins and UCPs in slowly developing diseases of the white matter of the brain as multiple sclerosis and leukoaraiosis.

  11. Hydroxynonenal, a lipid peroxidation end product, stimulates uncoupling protein activity in Acanthamoeba castellanii mitochondria; the sensitivity of the inducible activity to purine nucleotides depends on the membranous ubiquinone redox state.

    Science.gov (United States)

    Woyda-Ploszczyca, Andrzej M; Jarmuszkiewicz, Wieslawa

    2012-10-01

    We studied the influence of exogenously generated superoxide and exogenous 4-hydroxy-2-nonenal (HNE), a lipid peroxidation end product, on the activity of the Acanthamoeba castellanii uncoupling protein (AcUCP). The superoxide-generating xanthine/xanthine oxidase system was insufficient to induce mitochondrial uncoupling. In contrast, exogenously added HNE induced GTP-sensitive AcUCP-mediated mitochondrial uncoupling. In non-phosphorylating mitochondria, AcUCP activation by HNE was demonstrated by increased oxygen consumption accompanied by a decreased membrane potential and ubiquinone (Q) reduction level. The HNE-induced GTP-sensitive proton conductance was similar to that observed with linoleic acid. In phosphorylating mitochondria, the HNE-induced AcUCP-mediated uncoupling decreased the yield of oxidative phosphorylation. We demonstrated that the efficiency of GTP to inhibit HNE-induced AcUCP-mediated uncoupling was regulated by the endogenous Q redox state. A high Q reduction level activated AcUCP by relieving the inhibition caused by GTP while a low Q reduction level favoured the inhibition. We propose that the regulation of UCP activity involves a rapid response through the endogenous Q redox state that modulates the inhibition of UCP by purine nucleotides, followed by a late response through lipid peroxidation products resulting from an increase in the formation of reactive oxygen species that modulate the UCP activation.

  12. A novel intronic peroxisome proliferator-activated receptor gamma enhancer in the uncoupling protein (UCP) 3 gene as a regulator of both UCP2 and -3 expression in adipocytes

    DEFF Research Database (Denmark)

    Bugge, Anne Skovsø; Siersbaek, Majken; Madsen, Maria S

    2010-01-01

    Uncoupling Proteins (UCPs) are integral ion channels residing in the inner mitochondrial membrane. UCP2 is ubiquitously expressed, while UCP3 is found primarily in muscles and adipose tissue. Although the exact molecular mechanism of action is controversial, it is generally agreed that both...

  13. The developmentally regulated avian protein IFAPa-400 is transitin.

    Science.gov (United States)

    Ma, X; Charron, F; Cole, G J; Savard, P E; Vincent, M

    1998-07-01

    Transitin and IFAPa-400 are developmentally regulated high M(r) proteins expressed transiently in early chick embryogenesis. Both are associated with radially oriented fibers in the developing CNS and with various neural and myogenic tissues before their down-regulation at later stages. Previous studies have shown that IFAPa-400 colocalized and copurified with intermediate filament proteins and recent molecular cloning has indicated that transitin is a member of this family of cytoskeletal proteins. Here, we provide evidence that IFAPa-400 and transitin are the same protein. The sequence of a composite cDNA corresponding to more than 700 amino acids of IFAPa-400 carboxy-terminal extremity is identical to that of transitin. Both proteins exhibit identical apparent M(r) and isoelectric point. Immunopurified IFAPa-400 reacts with different antibodies to transitin and vice-versa. The patterns of expression of both proteins show a perfect coincidence at the tissue level. At the subcellular level, most antibodies to IFAPa-400/transitin decorate a typical intermediate filament network. However, monoclonal antibody A2B11, at the origin of transitin identification, exhibits a staining more typical of a cortical component, suggesting that different populations of transitin exist within the cell.

  14. Characterization of antigenic domains and epitopes in the ORF3 protein of a Chinese isolate of avian hepatitis E virus.

    Science.gov (United States)

    Zhao, Qin; Sun, Ya-ni; Hu, Shou-bin; Wang, Xin-jie; Xiao, Yi-hong; Hsu, Walter H; Xiao, Shu-qi; Wang, Cheng-bao; Mu, Yang; Hiscox, Julian A; Zhou, En-Min

    2013-12-27

    Avian hepatitis E virus (HEV) is an emerging virus associated with the big liver and spleen disease or hepatitis-splenomegaly syndrome in chickens and subclinical infections by the virus are also common. The complete genome of avian HEV contains three open-reading frames (ORFs) in which ORF2 protein is part of virus particles and thus contains primary epitopes. Antigenic epitopes of avian HEV ORF2 protein have been described but those associated with the ORF3 have not. To analyze the antigenic domains and epitopes in the ORF3 protein of a Chinese isolate of avian HEV (CaHEV), we generated a series of antigens comprised of the complete ORF3 and also five truncated overlapping ORF3 peptides. The antibodies used in this study were mouse antisera and monoclonal antibodies against ORF3, positive chicken sera from Specific Pathogen Free chickens experimentally infected with CaHEV and clinical chicken sera. Using these antigens and antibodies, we identified three antigenic domains at amino acids (aa) 1-28, 55-74 and 75-88 in which aa 75-88 was a dominant domain. The dominant domain contained at least two major epitopes since field chickens infected with avian HEV produced antibodies against the domain and epitopes. These results provide useful information for future development of immunoassays for the diagnosis of avian HEV infection.

  15. Spectrophotometric and Refractometric Determination of Total Protein in Avian Plasma

    OpenAIRE

    2013-01-01

    The aim of this study was to compare the total protein values obtained in heparin plasma of chickens by a spectrophotometric technique (biuret method), and the values obtained on the same day in the same samples by refractometry. The results obtained by refractometry (average value 2.638±0.153g%) were higher than those obtained by the spectrophotometric method (average value 2.441±0.181g%). There was a low correlation (r = 0.6709) between the total protein values, determined with both methods...

  16. Uncoupling protein 2 G(-866A polymorphism: a new gene polymorphism associated with C-reactive protein in type 2 diabetic patients C-reactive protein in type 2 diabetic patients

    Directory of Open Access Journals (Sweden)

    Cocozza Sergio

    2010-10-01

    Full Text Available Abstract Background This study evaluated the relationship between the G(-866A polymorphism of the uncoupling protein 2 (UCP2 gene and high-sensitivity C reactive protein (hs-CRP plasma levels in diabetic patients. Methods We studied 383 unrelated people with type 2 diabetes aged 40-70 years. Anthropometry, fasting lipids, glucose, HbA1c, and hs-CRP were measured. Participants were genotyped for the G (-866A polymorphism of the uncoupling protein 2 gene. Results Hs-CRP (mg/L increased progressively across the three genotype groups AA, AG, or GG, being respectively 3.0 ± 3.2, 3.6 ± 5.0, and 4.8 ± 5.3 (p for trend = 0.03. Since hs-CRP values were not significantly different between AA and AG genotype, these two groups were pooled for further analyses. Compared to participants with the AA/AG genotypes, homozygotes for the G allele (GG genotype had significantly higher hs-CRP levels (4.8 ± 5.3 vs 3.5 ± 4.7 mg/L, p = 0.01 and a larger proportion (53.9% vs 46.1%, p = 0.013 of elevated hs-CRP (> 2 mg/L. This was not explained by major confounders such as age, gender, BMI, waist circumference, HbA1c, smoking, or medications use which were comparable in the two genotype groups. Conclusions The study shows for the first time, in type 2 diabetic patients, a significant association of hs-CRP levels with the G(-866A polymorphism of UCP2 beyond the effect of major confounders.

  17. Detection of Markers of Increased Virulence Non Structural protein (NS I Avian Influenza Virus H5N1 from Indonesia=DETEKSI PENANDA PENINGKATAN VIRULENSI NON STRUKTURAL PROTEIN (NS1 VIRUS AVIAN INFLUENZA H5N1 ASAL INDONESIA

    Directory of Open Access Journals (Sweden)

    Arief Mulyono

    2015-03-01

    Full Text Available ENGLISHAbstractNS1 protein is a multifunction protein that plays key role of pathogenesis and virulence of avians influenza virus H5N1. The amino acid substitution at the position P42S, D92E, F103I, M106I and 5 amino acid deletion at the position 80 to 84 in NS1 protein reported increasing virulence of avians influenza virus H5N1. Several studies showed avians influenza virus H5N1 in Indonesia has dynamic changed. This study aimed to analyze the markers of virulence of NS1 protein sequences of all H5N1 virus isolates from Indonesia. The source of NS1 protein sequence data gene obtained from GeneBank and Gisaid. Data were analyzed using Bioedit software. The Results showed the isolates from Indonesia had substitutions P42S and 5 amino acids deletions at positions 80-84 resulting in the potential for increased virulence of the virus. However, amino acid substitution at the position D92E, F103L and M106I substitution were not found.INDONESIANAbstrakProtein NS1 adalah protein multifungsi yang memainkan peran kunci dalam patogenesis dan virulensi virus avian influenza H5N1. Substitusi asam amino P42S, D92E, F103I, M106I, dan delesi 5 asam amino di posisi 80 - 84 dilaporkan meningkatkan virulensi virus avian influenza H5N1. Beberapa penelitian menunjukkan bahwa virus avian influenza di Indonesia mengalami perubahan dinamis. Studi ini akan menganalisis motif asam amino yang menjadi penanda peningkatan virulensi pada sekuen protein NS1 virus avian influenza H5N1 asal Indonesia. Data sekuen asam amino protein NS1 diperoleh dari database GeneBank dan Gisaid. Analisis data menggunakan Bioedit software. Hasil analisis menunjukkan subtitusi asam amino dari prolin ke serin di posisi 42 (P42S dan delesi 5 asam amino di posisi 80 – 84 telah ditemukan pada virus avian influenza asal Indonesia, akan tetapi tidak ditemukan substitusi asam amino aspartat ke glutamat diposisi no 92 (D92E dan tidak ada yang mengalami 2 substitusi asam amino sekaligus diposisi 103

  18. Cloning, expression and immunogenicity of the avian pneumovirus (Colorado isolate) F protein.

    Science.gov (United States)

    Tarpey, I; Huggins, M B; Davis, P J; Shilleto, R; Orbell, S J; Cook, J K

    2001-10-01

    The F protein of the Colorado isolate of avian pneumovirus (APV), expressed from a DNA plasmid, was recognized by antiserum to both A and B subgroup APVs. After two intramuscular injections of turkeys with this plasmid, a homologous antibody response was detected by enzyme-linked immunosorbent assay. This antibody also recognized subgroup A APV. However, there was no neutralization of the Colorado isolate or of subgroup A or B viruses. Although no significant clinical protection was detected following homologous challenge of poults, an anamnestic serological response was seen, suggesting that a systemic antibody response but no local mucosal immunity was induced.

  19. The Effect of Resveratrol and Quercetin Treatment on PPAR Mediated Uncoupling Protein (UCP- 1, 2, and 3 Expression in Visceral White Adipose Tissue from Metabolic Syndrome Rats

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    Vicente Castrejón-Tellez

    2016-07-01

    Full Text Available Uncoupling proteins (UCPs are members of the mitochondrial anion carrier superfamily involved in the control of body temperature and energy balance regulation. They are currently proposed as therapeutic targets for treating obesity and metabolic syndrome (MetS. We studied the gene expression regulation of UCP1, -2, and -3 in abdominal white adipose tissue (WAT from control and MetS rats treated with two doses of a commercial mixture of resveratrol (RSV and quercetin (QRC. We found that UCP2 was the predominantly expressed isoform, UCP3 was present at very low levels, and UCP1 was undetectable. The treatment with RSV + QRC did not modify UCP3 levels; however, it significantly increased UCP2 mRNA in control and MetS rats in association with an increase in oleic and linoleic fatty acids. WAT from MetS rats showed a significantly increased expression of peroxisome proliferator-activated receptor (PPAR-α and PPAR-γ when compared to the control group. Furthermore, PPAR-α protein levels were increased by the highest dose of RSV + QRC in the control and MetS groups. PPAR-γ expression was only increased in the control group. We conclude that the RSV + QRC treatment leads to overexpression of UCP2, which is associated with an increase in MUFA and PUFA, which might increase PPAR-α expression.

  20. Epitope Mapping of Avian Influenza M2e Protein: Different Species Recognise Various Epitopes.

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    Noor Haliza Hasan

    Full Text Available A common approach for developing diagnostic tests for influenza virus detection is the use of mouse or rabbit monoclonal and/or polyclonal antibodies against a target antigen of the virus. However, comparative mapping of the target antigen using antibodies from different animal sources has not been evaluated before. This is important because identification of antigenic determinants of the target antigen in different species plays a central role to ensure the efficiency of a diagnostic test, such as competitive ELISA or immunohistochemistry-based tests. Interest in the matrix 2 ectodomain (M2e protein of avian influenza virus (AIV as a candidate for a universal vaccine and also as a marker for detection of virus infection in vaccinated animals (DIVA is the rationale for the selection of this protein for comparative mapping evaluation. This study aimed to map the epitopes of the M2e protein of avian influenza virus H5N1 using chicken, mouse and rabbit monoclonal or monospecific antibodies. Our findings revealed that rabbit antibodies (rAbs recognized epitope 6EVETPTRN13 of the M2e, located at the N-terminal of the protein, while mouse (mAb and chicken antibodies (cAbs recognized epitope 10PTRNEWECK18, located at the centre region of the protein. The findings highlighted the difference between the M2e antigenic determinants recognized by different species that emphasized the importance of comparative mapping of antibody reactivity from different animals to the same antigen, especially in the case of multi-host infectious agents such as influenza. The findings are of importance for antigenic mapping, as well as diagnostic test and vaccine development.

  1. Protective Effect of Peroxisome Proliferator-Activated Receptor α Activation against Cardiac Ischemia-Reperfusion Injury Is Related to Upregulation of Uncoupling Protein-3

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    Jong Wook Song

    2016-01-01

    Full Text Available Activation of peroxisome proliferator-activated receptor α (PPARα confers cardioprotection, while its mechanism remains elusive. We investigated the protective effect of PPARα activation against cardiac ischemia-reperfusion injury in terms of the expression of uncoupling protein (UCP. Myocardial infarct size and UCP expression were measured in rats treated with WY-14643 20 mg/kg, a PPARα ligand, or vehicle. WY-14643 increased UCP3 expression in vivo. Myocardial infarct size was decreased in the WY-14643 group (76 ± 8% versus 42 ± 12%, P<0.05. During reperfusion, the incidence of arrhythmia was higher in the control group compared with the WY-14643 group (9/10 versus 3/10, P<0.05. H9c2 cells were incubated for 24 h with WY-14643 or vehicle. WY-14643 increased UCP3 expression in H9c2 cells. WY-14643 decreased hypoxia-stimulated ROS production. Cells treated with WY-14643 were more resistant to hypoxia-reoxygenation than the untreated cells. Knocking-down UCP3 by siRNA prevented WY-14643 from attenuating the production of ROS. UCP3 siRNA abolished the effect of WY-14643 on cell viability against hypoxia-reoxygenation. In summary, administration of PPARα agonist WY-14643 mitigated the extent of myocardial infarction and incidence of reperfusion-induced arrhythmia. PPARα activation conferred cytoprotective effect against hypoxia-reoxygenation. Associated mechanisms involved increased UCP3 expression and resultant attenuation of ROS production.

  2. The Hydroxyl at Position C1 of Genipin Is the Active Inhibitory Group that Affects Mitochondrial Uncoupling Protein 2 in Panc-1 Cells.

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    Yang Yang

    Full Text Available Genipin (GNP effectively inhibits uncoupling protein 2 (UCP2, which regulates the leakage of protons across the inner mitochondrial membrane. UCP2 inhibition may induce pancreatic adenocarcinoma cell death by increasing reactive oxygen species (ROS levels. In this study, the hydroxyls at positions C10 (10-OH and C1 (1-OH of GNP were hypothesized to be the active groups that cause these inhibitory effects. Four GNP derivatives in which the hydroxyl at position C10 or C1 was replaced with other chemical groups were synthesized and isolated. Differences in the inhibitory effects of GNP and its four derivatives on pancreatic carcinoma cell (Panc-1 proliferation were assessed. The effects of GNP and its derivatives on apoptosis, UCP2 inhibition and ROS production were also studied to explore the relationship between GNP's activity and its structure. The derivatives with 1-OH substitutions, geniposide (1-GNP1 and 1-ethyl-genipin (1-GNP2 lacked cytotoxic effects, while the other derivatives that retained 1-OH, 10-piv-genipin (10-GNP1 and 10-acetic acid-genipin (10-GNP2 exerted biological effects similar to those of GNP, even in the absence of 10-OH. Thus, 1-OH is the key functional group in the structure of GNP that is responsible for GNP's apoptotic effects. These cytotoxic effects involve the induction of Panc-1 cell apoptosis through UCP2 inhibition and subsequent ROS production.

  3. Interleukin-22 restored mitochondrial damage and impaired glucose-stimulated insulin secretion through down-regulation of uncoupling protein-2 in INS-1 cells.

    Science.gov (United States)

    Hu, Minling; Lin, Hanxiao; Yang, Li; Cheng, Yanzhen; Zhang, Hua

    2017-01-07

    Defective glucose-stimulated insulin secretion (GSIS) induced by chronic exposure to fatty acids is a hallmark of type 2 diabetes (T2D). Interleukin-22 (IL-22) has been shown to exert beneficial effects on insulin secretion and to protect pancreatic β-cells from stress. Moreover, uncoupling protein-2 (UCP-2) plays a central role in the regulation of GSIS and β-cell dysfunction, whereas the role of UCP-2 in IL-22-enhanced glycemic control under conditions of lipotoxicity remains unclear. In this present study, we investigated the effects of IL-22 on rat insulin-secreting cells (INS-1 cells) and the mechanisms that underlie IL-22 and lipotoxicity-impaired GSIS in vitro. Chronic palmitate (PA) treatment impaired insulin secretion and activated UCP-2 expression in INS-1 cells. Furthermore, in INS-1 cells, both reduced mitochondrial membrane potential (ΔΨm) and impaired GSIS induced by PA treatment were effectively reversed by an inhibitor of UCP-2 (genipin). Additionally, compared with the PA-treated group, INS-1 cells treated with IL-22 down-regulated UCP-2 expression, increased mitochondrial membrane potential, and restored GSIS. Together, our findings indicate that chronic exposure to PA could activate UCP-2, resulting in mitochondrial damage and impaired GSIS in INS-1 cells. We also suggest that IL-22 plays a protective role in this process via the down-regulation of UCP-2.

  4. Uncoupling Protein 2 (UCP2) Function in the Brain as Revealed by the Cerebral Metabolism of (1-(13)C)-Glucose.

    Science.gov (United States)

    Contreras, Laura; Rial, Eduardo; Cerdan, Sebastian; Satrustegui, Jorgina

    2017-01-01

    The mitochondrial aspartate/glutamate transporter Aralar/AGC1/Slc25a12 is critically involved in brain aspartate synthesis, and AGC1 deficiency results in a drastic fall of brain aspartate levels in humans and mice. It has recently been described that the uncoupling protein UCP2 transports four carbon metabolites including aspartate. Since UCP2 is expressed in several brain cell types and AGC1 is mainly neuronal, we set to test whether UCP2 could be a mitochondrial aspartate carrier in the brain glial compartment. The study of the cerebral metabolism of (1-(13)C)-glucose in vivo in wild type and UCP2-knockout mice showed no differences in C3 or C2 labeling of aspartate, suggesting that UCP2 does not function as a mitochondrial aspartate carrier in brain. However, surprisingly, a clear decrease (of about 30-35 %) in the fractional enrichment of glutamate, glutamine and GABA was observed in the brains of UCP2-KO mice which was not associated with differences in either glucose or lactate enrichments. The results suggest that the dilution in the labeling of glutamate and its downstream metabolites could originate from the uptake of an unlabeled substrate that could not leave the matrix via UCP2 becoming trapped in the matrix. Understanding the nature of the unlabeled substrate and its precursor(s) as alternative substrates to glucose is of interest in the context of neurological diseases associated with UCP2.

  5. Genetic evolution analysis of matrix protein 2 gene of avian influenza H5N1 viruses from boundary of Yunnan province

    Institute of Scientific and Technical Information of China (English)

    肖雪

    2013-01-01

    Objective To elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein (M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province

  6. Generation and characterisation of monoclonal antibodies specific to avian influenza H7N9 haemagglutinin protein

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    A Malik

    2016-01-01

    Full Text Available Introduction: Emerging virulent strains of influenza virus pose a serious public health threat with potential pandemic consequences. A novel avian influenza virus, H7N9, breached the species barrier from infected domestic poultry to humans in 2013 in China. Since then, it has caused numerous infections in humans with a close contact to poultry. Materials and Methods: In this study, we describe the preliminary characterisation of five murine monoclonal antibodies (MAbs developed against recombinant haemagglutinin (rHA protein of avian H7N9 A/Anhui/1/2013 virus by their Western blot and enzyme-linked immunosorbent assay (ELISA reactivity and binding affinity. Results: Of the five MAbs, four were highly specific to H7N9 HA and did not show any cross-reactivity in ELISA with rHA protein from pandemic as well as seasonal H1N1, H2N2, H3N2, H5N1 and influenza virus B (B/Brisbane/60/2008. However, one of the MAbs, MA-24, in addition to HA protein of H7N9 also reacted strongly with HA protein of H3N2 and weakly with HA of pandemic and seasonal H1N1 and H2N2. All the five MAbs also reacted with H7N9 rHA in Western blot. The MAbs bound H7N9 rHA with an equilibrium dissociation constant (KD ranging between 0.14 and 25.20 nM, indicating their high affinity to HA. Conclusions: These antibodies may be useful in developing diagnostic tools for the detection of influenza H7N9 virus infections.

  7. A Novel Intronic Peroxisome Proliferator-activated Receptor γ Enhancer in the Uncoupling Protein (UCP) 3 Gene as a Regulator of Both UCP2 and -3 Expression in Adipocytes*

    OpenAIRE

    2010-01-01

    Uncoupling Proteins (UCPs) are integral ion channels residing in the inner mitochondrial membrane. UCP2 is ubiquitously expressed, while UCP3 is found primarily in muscles and adipose tissue. Although the exact molecular mechanism of action is controversial, it is generally agreed that both homologues function to facilitate mitochondrial fatty acid oxidation. UCP2 and -3 expression is activated by the peroxisome proliferator-activated receptors (PPARs), but so far no PPAR response element has...

  8. The Preservation of in vivo Phosphorylated and Activated Uncoupling Protein 3 (UCP3) in Isolated Skeletal Muscle Mitochondria following Administration of 3,4-Methylenedioxymethamphetamine (MDMA aka Ecstasy) to Rats/Mice

    OpenAIRE

    2012-01-01

    PUBLISHED Previous researchers have demonstrated that 3,4-methylenedioxymethamphetamine (MDMA) induced hyperthermia, in skeletal muscle of animals, is uncoupling protein 3 (UCP3) dependent. In light of our investigations that in vivo phosphorylation of UCP1 is augmented under conditions of cold-acclimation, we set out to investigate whether (a) UCP3 was phosphorylated in vivo and (b) whether in vivo phosphorylation of UCP3 resulted in increased proton leak following MDMA administration to ...

  9. Skeletal muscle uncoupling protein-3 restores upon intervention in the pre-diabetic and diabetic state: implications for diabetes pathogenesis?

    NARCIS (Netherlands)

    Mensink, M.; Hesselink, M.K.C.; Borghouts, L.B.; Keizer, H.A.; Moonen-Kornips, E.; Schaart,; Blaak,; Schrauwen,

    2007-01-01

    Both exercise training and a lifestyle-intervention program increase UCP3 protein content inskeletal muscle of subjects with reduced glycaemic control, indicating a restoration towards normal UCP3 levels. These data support the idea that UCP3 has a role in the aetiology of type 2 diabetes mellitus

  10. Effects of Berberine on Hepatic Sirtuin 1-uncoupling Protein 2 Pathway in Non-alcoholic Fatty Liver Disease Rats Induced by High-fat Diet

    Institute of Scientific and Technical Information of China (English)

    Yuan-jun Deng; Yu-pei Zhang; Qin-he Yang; Li Han; Yin-ji Liang; Yi-fang He; Yuan-yuan Li

    2016-01-01

    Objective To investigate the involvement of sirtuin 1 (SIRT1)-uncoupling protein 2 (UCP2) pathway in the development of non-alcoholic fatty liver disease and whether berberine exerts its effects by regulating this pathway.Methods Male SD rats were divided into three groups:normal control group,high-fat diet group,and berberine supplement group.The rats in the normal control group were given normal diet while the rats in the other two groups were fed with high-fat diet.Rats in the berberine supplement group were concurrently given berberine (100 mg/kg body weight) once daily.After 1 6 weeks,the levels of serum,liver lipids,and serum aminotransferase were measured using an automatic biochemical analyzer.Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the liver were measured using commercial kits.Histopathological changes of liver tissues were observed by hematoxylin and eosin (HE) staining and Oil Red O staining.The hepatic mRNA and protein levels of SIRT1 and UCP2 were assayed by reverse transcription polymerase chain reaction (RT-PCR) or Western blotting.Results Berberine supplement could significantly decrease the serum and liver lipid contents in rats fed with high-fat diet.Meanwhile,SOD level was significantly elevated,but MDA level was reduced in the liver.The results of HE and Oil Red O staining showed that the hepatic steatosis was alleviated in berberine supplement group.Furthermore,berberine induced an increase in SlRT1 expression but a decrease in UCP2 expression.Conclusion The regulation of hepatic SIRT1-UCP2 pathway may be an important mechanism by which berberine exerts the beneficial effects in NAFLD rats.

  11. Development and evaluation of an immunochromatographic strip for rapid detection of capsid protein antigen p27 of avian leukosis virus.

    Science.gov (United States)

    Qian, Kun; Liang, You-zhi; Yin, Li-ping; Shao, Hong-xia; Ye, Jian-qiang; Qin, Ai-jian

    2015-09-01

    A rapid immunochromatographic strip for detecting capsid protein antigen p27 of avian leukosis virus was successfully developed based on two high-affinity monoclonal antibodies. The test strip could detect not only 600pg purified recombinant p27 protein but also quantified avian leukosis virus as low as 70 TCID50, which has comparative sensitivity to the commercial enzyme-linked immunosorbent assay (ELISA) kit. For the evaluation of this test strip, 1100 samples consisting of cloacal swabs, meconium collected from the earliest stool of one day old chicken and virus isolates were assessed both by the strip and by the commercial ELISA kit. The agreement between these two tests was 93.91%, 93.42% and 100%, respectively. The sensitivity and specificity of the strip were also calculated by using the ELISA kit as the standard. This immunochromatographic strip provides advantages of rapid and simple detection of capsid protein antigen p27 of avian leukosis virus, which could be applied as an on-site testing assay and used for control and eradication programs of avian leukosis disease.

  12. Expression of recombinant small hydrophobic protein for serospecific detection of avian pneumovirus subgroup C.

    Science.gov (United States)

    Luo, Lizhong; Sabara, Marta I; Li, Yan

    2005-01-01

    The small hydrophobic (SH) gene of the avian pneumovirus (APV) Colorado isolate (CO), which belongs to subgroup C (APV/C), was expressed with a baculovirus vector. The recombinant SH protein was evaluated as a potential subgroup-specific diagnostic reagent in order to differentiate infections resulting from APV/C from those induced by APV/A, APV/B, and human metapneumovirus (hMPV). When the recombinant baculovirus was used to infect insect cells, a 31- to 38-kDa glycosylated form of the SH protein was produced and subsequently tested for reactivity with antibodies specific for APV/A, APV/B, APV/C, and hMPV. Western blot analysis showed that the expressed recombinant SH protein could only be recognized by APV/C-specific antibodies. This result was consistent with sequence analysis of the APV/C SH protein, which had very low (24%) amino acid identity with the corresponding protein of hMPV and no discernible identity with the SH protein of APV/A or APV/B. A recombinant SH protein-based enzyme-linked immunosorbent assay (ELISA) was developed, and it further confirmed the lack of reactivity of this protein with antisera raised to APV/A, APV/B, and hMPV and supported its designation as a subgroup-specific antigen. This finding indicated that the recombinant SH protein was a suitable antigen for ELISA-based detection of subgroup-specific antibodies in turkeys and could be used for serologically based differential diagnosis of APV and hMPV infections.

  13. Expression and regulation of the decoy bone morphogenetic protein receptor BAMBI in the developing avian face.

    Science.gov (United States)

    Higashihori, Norihisa; Song, Yiping; Richman, Joy M

    2008-05-01

    Here, we examine the expression and regulation of the gene BAMBI, a kinase-deficient decoy receptor capable of interacting with type I bone morphogenetic protein (BMP) receptors in avian embryos. Initially, expression was limited to the endoderm during neurula and pharyngula stages. From embryonic day 3.5 (stage 20) and onward, BAMBI expression almost perfectly overlapped with known expression patterns for BMP4, particularly in the face and limbs. We performed bead implant experiments in the face to see which signals could be repressing or promoting expression of BAMBI. Our data point to retinoids and BMPs as being major positive regulators of BAMBI expression; however, fibroblast growth factor 2 acts to repress BAMBI. Furthermore, retinoic acid is likely to act directly on BAMBI as induction occurs in the presence of cycloheximide. The data suggested that BAMBI could be used to regulate Bmp signaling during tissue interactions that are an integral part of facial morphogenesis.

  14. Production and purification of avian antibodies (IgYs from inclusion bodies of a recombinant protein central in NAD+ metabolism

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    Paula A. Moreno-González

    2013-08-01

    Full Text Available The use of hens for the production of polyclonal antibodies reduces animal intervention and moreover yields a higher quantity of antibodies than other animal models.  The phylogenetic distance between bird and mammal antigens, often leads to more specific avian antibodies than their mammalian counterparts.Since a large amount of antigen is required for avian antibody production, the use of recombinant proteins for this procedure has been growing faster over the last years. Nevertheless, recombinant protein production through heterologous systems frequently prompts the protein to precipitate, forming insoluble aggregates of limited utility (inclusion bodies. A methodology for the production of avian polyclonal antibodies, using recombinant protein from inclusion bodies is presented in this article.In order to produce the antigen, a recombinant Nicotinamide mononucleotide adenylyltransferase from Giardia intestinalis (His-GiNMNAT was expressed in Escherichia coli.  The protein was purified through solubilization from inclusion bodies prior to its renaturalization.  Antibodies were purified from egg yolk of immunized hens by water dilution, followed by ammonium sulfate precipitation and thiophilic affinity chromatography.The purified antibodies were tested against His-GiNMNAT protein in Western blot essays. From one egg yolk, 14.4 mg of highly pure IgY were obtained; this antibody was able to detect 15ng of His-GiNMNAT.  IgY specificity was improved by means of antigen affinity purification, allowing its use for parasite protein recognition.

  15. Generation and Characterization of Monoclonal Antibodies Specific to Avian Influenza H5N1 Hemagglutinin Protein.

    Science.gov (United States)

    Malik, Ankita; Mallajosyula, V Vamsee Aditya; Mishra, Nripendra Nath; Varadarajan, Raghavan; Gupta, Satish Kumar

    2015-12-01

    Highly pathogenic avian influenza (HPAI) H5N1 virus has in the past breached the species barrier from infected domestic poultry to humans in close contact. Although human-to-human transmission has previously not been reported, HPAI H5N1 virus has pandemic potential owing to gain of function mutation(s) and/or genetic reassortment with human influenza A viruses. Monoclonal antibodies (MAbs) have been used for diagnosis as well as specific therapeutic candidates in several disease conditions including viral infections in humans. In this study, we describe the preliminary characterization of four murine MAbs developed against recombinant hemagglutinin (rHA) protein of avian H5N1 A/turkey/Turkey/1/2005 virus that are either highly specific or broadly reactive against HA from other H5N1 subtype viruses, such as A/Hong Kong/213/03, A/Common magpie/Hong Kong/2256/2006, and A/Barheaded goose/Quinghai/14/2008. The antibody binding is specific to H5N1 HAs, as none of the antibodies bound H1N1, H2N2, H3N2, or B/Brisbane/60/2008 HAs. Out of the four MAbs, one of them (MA-7) also reacted weakly with the rHA protein of H7N9 A/Anhui/1/2013. All four MAbs bound H5 HA (A/turkey/Turkey/1/2005) with high affinity with an equilibrium dissociation constant (KD) ranging between 0.05 and 10.30 nM. One of the MAbs (MA-1) also showed hemagglutination inhibition activity (HI titer; 31.25 μg/mL) against the homologous A/turkey/Turkey/1/2005 H5N1 virus. These antibodies may be useful in developing diagnostic tools for detection of influenza H5N1 virus infection.

  16. 解偶联蛋白与肥胖及运动的关系%Relation of uncoupling proteins with obesity and exercise

    Institute of Scientific and Technical Information of China (English)

    张慧敏; 程旭光

    2005-01-01

    目的:解偶联蛋白 (Uncoupling proteins,UCPS)可以驱散质子电化学梯度,使呼吸链和 ATP的合成解偶联,从而影响产热和能量代谢.目前已发现 5种 UCPS家族成员,它们之间具有不同程度的同源性. UCPS现已被列为肥胖的候选基因.通过了解 UCPS的作用机制、 UCPS多态性与肥胖的关系及其表达的调节因素等,可以为肥胖发生机制及其防治工作的研究开辟新途径. 资料来源:应用计算机检索 1994/2004Medline的相关文章,检索词" Uncoupling proteins",并限定文章语言种类为英文.同时计算机检索 1994/2004中国期刊全文数据库( http://www.sy.cnki.net)的相关文章,限定文章语言种类为中文,检索项"关键词",检索词"解偶联蛋白",选择医药卫生辑专栏目录.另外手工检索到相关文献 5篇. 资料选择:对资料进行初审,被选文献符合以下标准:①解偶联蛋白家族中各成员研究.②解偶联蛋白与肥胖方面的研究.③解偶联蛋白与运动关系的研究. 资料提炼:共收集到 35篇相关文献,中文文献均为全文,部分外文文献为全文,其他为摘要.在这些文献中符合标准的有 22篇. 资料综合: UCP1在人体能量平衡中的作用不大. UCP2和 UCP3与肥胖和 2型糖尿病的发生可能有一定的相关性. UCP4基因表达于大鼠脂肪组织中,并可能参与肥胖的发生发展.摄食状况是影响 UCPs基因表达的重要因素.一次性运动可以不同水平的上调 UCP3 mRNA水平,这可能与训练水平、运动方式和运动持续时间不同相关.而长期耐力训练却降低了 UCPs基因的表达. 结论: UCPs与肥胖有一定的关系,可能是防治肥胖的一个有效的靶位点.运动可以不同程度影响 UCPs基因的表达,这与肥胖也有一定的关系.

  17. Disruption of the mitochondrial alternative oxidase (AOX) and uncoupling protein (UCP) alters rates of foliar nitrate and carbon assimilation in Arabidopsis thaliana.

    Science.gov (United States)

    Gandin, Anthony; Denysyuk, Mykhaylo; Cousins, Asaph B

    2014-07-01

    Under high light, the rates of photosynthetic CO2 assimilation can be influenced by reductant consumed by both foliar nitrate assimilation and mitochondrial alternative electron transport (mAET). Additionally, nitrate assimilation is dependent on reductant and carbon skeletons generated from both the chloroplast and mitochondria. However, it remains unclear how nitrate assimilation and mAET coordinate and contribute to photosynthesis. Here, hydroponically grown Arabidopsis thaliana T-DNA insertional mutants for alternative oxidase (AOX1A) and uncoupling protein (UCP1) fed either NO3 (-) or NH4 (+) were used to determine (i) the response of NO3 (-) uptake and assimilation to the disruption of mAET, and (ii) the interaction of N source (NO3 (-) versus NH4 (+)) and mAET on photosynthetic CO2 assimilation and electron transport. The results showed that foliar NO3 (-) assimilation was enhanced in both aox1a and ucp1 compared with the wild-type, suggesting that foliar NO3 (-) assimilation is probably driven by a decreased capacity of mAET and an increase in reductant within the cytosol. Wild-type plants had also higher rates of net CO2 assimilation (A net) and quantum yield of PSII (ϕPSII) under NO3 (-) feeding compared with NH4 (+) feeding. Additionally, under NO3 (-) feeding, A net and ϕPSII were decreased in aox1a and ucp1 compared with the wild type; however, under NH4 (+) they were not significantly different between genotypes. This indicates that NO3 (-) assimilation and mAET are both important to maintain optimal rates of photosynthesis, probably in regulating reductant accumulation and over-reduction of the chloroplastic electron transport chain. These results highlight the importance of mAET in partitioning energy between foliar nitrogen and carbon assimilation.

  18. Altered expression of uncoupling protein 2 in GLP-1-producing cells after chronic high glucose exposure: implications for the pathogenesis of diabetes mellitus.

    Science.gov (United States)

    Urbano, Francesca; Filippello, Agnese; Di Pino, Antonino; Barbagallo, Davide; Di Mauro, Stefania; Pappalardo, Alessandro; Rabuazzo, Agata Maria; Purrello, Michele; Purrello, Francesco; Piro, Salvatore

    2016-04-01

    Glucagon-like peptide-1 (GLP-1) is a gut L-cell hormone that enhances glucose-stimulated insulin secretion. Several approaches that prevent GLP-1 degradation or activate the GLP-1 receptor are being used to treat type 2 diabetes mellitus (T2DM) patients. In T2DM, GLP-1 secretion has been suggested to be impaired, and this defect appears to be a consequence rather than a cause of impaired glucose homeostasis. However, although defective GLP-1 secretion has been correlated with insulin resistance, little is known about the direct effects of chronic high glucose concentrations, which are typical in diabetes patients, on GLP-1-secreting cell function. In the present study, we demonstrate that glucotoxicity directly affects GLP-1 secretion in GLUTag cells chronically exposed to high glucose. Our results indicate that this abnormality is associated with a decrease in ATP production due to the elevated expression of mitochondrial uncoupling protein 2 (UCP2). Furthermore, UCP2 inhibition using small interfering RNA (siRNA) and the application of glibenclamide, an ATP-sensitive potassium (KATP(+)) channel blocker, reverse the GLP-1 secretion defect induced by chronic high-glucose treatment. These results show that glucotoxicity diminishes the secretory responsiveness of GLP-1-secreting cells to acute glucose stimulation. We conclude that the loss of the incretin effect, as observed in T2DM patients, could at least partially depend on hyperglycemia, which is typical in diabetes patients. Such an understanding may not only provide new insight into diabetes complications but also ultimately contribute to the identification of novel molecular targets within intestinal L-cells for controlling and improving endogenous GLP-1 secretion.

  19. Meta-analysis reveals the association of common variants in the uncoupling protein (UCP 1-3 genes with body mass index variability.

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    Letícia A Brondani

    Full Text Available BACKGROUND: The relationship between uncoupling protein (UCP 1-3 polymorphisms and susceptibility to obesity has been investigated in several genetic studies. However, the impact of these polymorphisms on obesity is still under debate, with contradictory results being reported. Until this date, no meta-analysis evaluated the association of UCP polymorphisms with body mass index (BMI variability. Thus, this paper describe a meta-analysis conducted to evaluate if the -3826A/G (UCP1; -866G/A, Ala55Val and Ins/Del (UCP2 and -55C/T (UCP3 polymorphisms are associated with BMI changes. METHODS: A literature search was run to identify all studies that investigated associations between UCP1-3 polymorphisms and BMI. Weighted mean differences (WMD were calculated for different inheritance models. RESULTS: Fifty-six studies were eligible for inclusion in the meta-analysis. Meta-analysis results showed that UCP2 55Val/Val genotype was associated with increased BMI in Europeans [Random Effect Model (REM WMD 0.81, 95% CI 0.20, 1.41]. Moreover, the UCP2 Ins allele and UCP3-55T/T genotype were associated with increased BMI in Asians [REM WMD 0.46, 95% CI 0.09, 0.83 and Fixed Effect Model (FEM WMD 1.63, 95% CI 0.25, 3.01]. However, a decreased BMI mean was observed for the UCP2-866 A allele in Europeans under a dominant model of inheritance (REM WMD -0.18, 95% CI -0.35, -0.01. There was no significant association of the UCP1-3826A/G polymorphism with BMI mean differences. CONCLUSIONS: The meta-analysis detected a significant association between the UCP2-866G/A, Ins/Del, Ala55Val and UCP3-55C/T polymorphisms and BMI mean differences.

  20. MiR-133a Is Functionally Involved in Doxorubicin-Resistance in Breast Cancer Cells MCF-7 via Its Regulation of the Expression of Uncoupling Protein 2.

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    Yuan Yuan

    Full Text Available The development of novel targeted therapies holds promise for conquering chemotherapy resistance, which is one of the major hurdles in current breast cancer treatment. Previous studies indicate that mitochondria uncoupling protein 2 (UCP-2 is involved in the development of chemotherapy resistance in colon cancer and lung cancer cells. In the present study we found that lower level of miR133a is accompanied by increased expression of UCP-2 in Doxorubicin-resistant breast cancer cell cline MCF-7/Dox as compared with its parental cell line MCF-7. We postulated that miR133a might play a functional role in the development of Doxorubicin-resistant in breast cancer cells. In this study we showed that: 1 exogenous expression of miR133a in MCF-7/Dox cells can sensitize their reaction to the treatment of Doxorubicin, which is coincided with reduced expression of UCP-2; 2 knockdown of UCP-2 in MCF-7/Dox cells can also sensitize their reaction to the treatment of Doxorubicin; 3 intratumoral delivering of miR133a can restore Doxorubicin treatment response in Doxorubicin-resistant xenografts in vivo, which is concomitant with the decreased expression of UCP-2. These findings provided direct evidences that the miR133a/UCP-2 axis might play an essential role in the development of Doxorubicin-resistance in breast cancer cells, suggesting that the miR133a/UCP-2 signaling cohort could be served as a novel therapeutic target for the treatment of chemotherapy resistant in breast cancer.

  1. Alteration in cardiac uncoupling proteins and eNOS gene expression following high-intensity interval training in favor of increasing mechanical efficiency

    Science.gov (United States)

    Fallahi, Ali Asghar; Shekarfroush, Shahnaz; Rahimi, Mostafa; Jalali, Amirhossain; Khoshbaten, Ali

    2016-01-01

    Objective(s): High-intensity interval training (HIIT) increases energy expenditure and mechanical energy efficiency. Although both uncoupling proteins (UCPs) and endothelial nitric oxide synthase (eNOS) affect the mechanical efficiency and antioxidant capacity, their effects are inverse. The aim of this study was to determine whether the alterations of cardiac UCP2, UCP3, and eNOS mRNA expression following HIIT are in favor of increased mechanical efficiency or decreased oxidative stress. Materials and Methods: Wistar rats were divided into five groups: control group (n=12), HIIT for an acute bout (AT1), short term HIIT for 3 and 5 sessions (ST3 and ST5), long-term training for 8 weeks (LT) (6 in each group). The rats of the training groups were made to run on a treadmill for 60 min in three stages: 6 min running for warm-up, 7 intervals of 7 min running on treadmill with a slope of 5° to 20° (4 min with an intensity of 80-110% VO2max and 3 min at 50-60% VO2max), and 5-min running for cool-down. The control group did not participate in any exercise program. Rats were sacrificed and the hearts were extracted to analyze the levels of UCP2, UCP3 and eNOS mRNA by RT-PCR. Results: UCP3 expression was increased significantly following an acute training bout. Repeated HIIT for 8 weeks resulted in a significant decrease in UCPs mRNA and a significant increase in eNOS expression in cardiac muscle. Conclusion: This study indicates that Long term HIIT through decreasing UCPs mRNA and increasing eNOS mRNA expression may enhance energy efficiency and physical performance. PMID:27114795

  2. [Effects of palmitic acid on activity of uncoupling proteins and proton leak in in vitro cerebral mitochondria from the rats exposed to simulated high altitude hypoxia].

    Science.gov (United States)

    Xu, Yu; Liu, Jun-Ze; Xia, Chen

    2008-02-25

    To reveal the roles of uncoupling proteins (UCPs) in disorder of mitochondrial oxidative phosphorylation induced by free fatty acid during hypoxic exposure, the effects of palmitic acid on activity of UCPs, proton leak and mitochondrial membrane potential in hypoxia-exposed rat brain mitochondria were observed in vitro. Adult Sprague-Dawley (SD) rats were set randomly into control, acute hypoxia and chronic hypoxia groups (n=8 in each group). The acute and chronic hypoxic rats were exposed to simulated 5000 m high altitude in a hypobaric chamber 23 h/d for 3 d and 30 d, respectively. The brain mitochondria were isolated by centrifugation. UCP content and activity were detected by [(3)H]-GTP binding method. The proton leak was measured by TPMP(+) electrode and oxygen electrode. The membrane potential of mitochondria was calculated by detecting the fluorescence from Rodamine 123. Hypoxic exposure resulted in an increase in UCP activity and content as well as proton leak, but a decrease in the membrane potential of rat brain mitochondria. Palmitic acid resulted in further increases in UCP activity and content as well as proton leak, and further decrease in membrane potential of brain mitochondria in vitro from hypoxia-exposed rats, but hypoxic exposure decreased the reactivity of cerebral mitochondria to palmitic acid, especially in the acute hypoxia group. There was a negative correlation between mitochondrial proton leak and K(d) value (representing derivative of UCP activity, PB(max) (representing the maximal content of UCPs in mitochondrial inner membrane, P<0.01, r = 0.856). Cerebral mitochondrial membrane potential was negatively correlated with proton leak (P<0.01, r = -0.880). It is suggested that hypoxia-induced proton leak enhancement and membrane potential decrease are correlated with the increased activity of UCPs. Hypoxia can also decrease the sensitivity of cerebral mitochondria to palmitic acid, which may be a self-protective mechanism in high altitude

  3. Recognition for avian influenza virus proteins based on support vector machine and linear discriminant analysis

    Institute of Scientific and Technical Information of China (English)

    LIANG GuiZhao; LIAO ChunYang; WU ShiRong; LI GenRong; HE Liu; GAO JianKun; Gan MengYu; LI DeJing; CHEN GuoPing; WANG GuiXue; LONG Sha; CHEN ZeCong; JING JuHua; ZHENG XiaoLin; ZENG Hui; ZHANG QiaoXia; ZHANG MengJun; YANG Qi; TIAN FeiFei; TONG JianBo; WANG JiaoNa; LIU YongHong; YANG ShanBin; LI Bo; QIU LiangJia; CAI ShaoXi; ZHAO Na; YANG Yan; SU XiaLi; SONG Jian; CHEN MeiXia; ZHANG XueJiao; SUN JiaYing; MEI Hu; LI JingWei; CHEN GuoHua; CHEN Gang; DENG Jie; PENG ChuanYou; ZHU WanPing; XU LuoNan; WU YuQuan; LIAO LiMin; LI Zhi; ZHOU Yuan; LI Jun; LU DaJun; SU QinLiang; HUANG ZhengHu; ZHOU Ping; LI ZhiLiang; YANG Li; ZHOU Peng; YANG ShengXi; SHU Mao

    2008-01-01

    Total 200 properties related to structural characteristics were employed to represent structures of 400 HA coded proteins of influenza virus as training samples.Some recognition models for HA proteins of avian influenza virus (AIV) were developed using support vector machine (SVM) and linear discriminant analysis (LDA).The results obtained from LDA are as follows: the identification accuracy (Ria) for training samples is 99.8% and Ria by leave one out cross validation is 99.5%.Both Ria of 99.8% for training samples and Ria of 99.3% by leave one out cross validation are obtained using SVM model, respectively.External 200 HA proteins of influenza virus were used to validate the external predictive power of the resulting model.The external Ria for them is 95.5% by LDA and 96.5% by SVM, respectively, which shows that HA proteins of AIVs are preferably recognized by SVM and LDA, and the performances by SVM are superior to those by LDA.

  4. Recognition for avian influenza virus proteins based on support vector machine and linear discriminant analysis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Total 200 properties related to structural characteristics were employed to represent structures of 400 HA coded proteins of influenza virus as training samples. Some recognition models for HA proteins of avian influenza virus (AIV) were developed using support vector machine (SVM) and linear discriminant analysis (LDA). The results obtained from LDA are as follows: the identification accuracy (Ria) for training samples is 99.8% and Ria by leave one out cross validation is 99.5%. Both Ria of 99.8% for training samples and Ria of 99.3% by leave one out cross validation are obtained using SVM model, respectively. External 200 HA proteins of influenza virus were used to validate the external predictive power of the resulting model. The external Ria for them is 95.5% by LDA and 96.5% by SVM, respectively, which shows that HA proteins of AIVs are preferably recognized by SVM and LDA, and the performances by SVM are superior to those by LDA.

  5. Identification of a linear B-cell epitope on the avian leukosis virus P27 protein using monoclonal antibodies.

    Science.gov (United States)

    Li, Xiaofei; Qin, Liting; Zhu, Haibo; Sun, Yingjun; Cui, Xuezhi; Gao, Yadong; Qi, Xiaole; Wang, Yongqiang; Gao, Honglei; Gao, Yulong; Wang, Xiaomei

    2016-10-01

    Avian leukosis virus (ALV) is an avian oncogenic retrovirus that can induce various clinical tumors. The capsid protein P27 is the group-specific antigen of ALV and has many viral antigen sites that are easy to detect. In this study, we produced a monoclonal antibody (mAb), 3A9, that is specific for the P27 protein. A series of partially overlapping peptides were screened to define (181)PPSAR(185) as the minimal linear epitope recognized by mAb 3A9. The identified epitope could be recognized by chicken anti-ALV and mouse anti-ALV P27 sera. The epitope was highly conserved among a number of ALV-A, ALV-B and ALV-J strains. MAb 3A9 might be a valuable tool for the development of new immunodiagnostic approaches for ALV, and the defined linear epitope might help further our understanding of the antigenic structure of the P27 protein.

  6. Molecular Characterizations of Surface Proteins Hemagglutinin and Neuraminidase from Recent H5Nx Avian Influenza Viruses

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Hua; Carney, Paul J.; Mishin, Vasiliy P.; Guo, Zhu; Chang, Jessie C.; Wentworth, David E.; Gubareva, Larisa V.; Stevens, James; Schultz-Cherry, S.

    2016-04-06

    ABSTRACT

    During 2014, a subclade 2.3.4.4 highly pathogenic avian influenza (HPAI) A(H5N8) virus caused poultry outbreaks around the world. In late 2014/early 2015, the virus was detected in wild birds in Canada and the United States, and these viruses also gave rise to reassortant progeny, composed of viral RNA segments (vRNAs) from both Eurasian and North American lineages. In particular, viruses were found with N1, N2, and N8 neuraminidase vRNAs, and these are collectively referred to as H5Nx viruses. In the United States, more than 48 million domestic birds have been affected. Here we present a detailed structural and biochemical analysis of the surface antigens of H5N1, H5N2, and H5N8 viruses in addition to those of a recent human H5N6 virus. Our results with recombinant hemagglutinin reveal that these viruses have a strict avian receptor binding preference, while recombinantly expressed neuraminidases are sensitive to FDA-approved and investigational antivirals. Although H5Nx viruses currently pose a low risk to humans, it is important to maintain surveillance of these circulating viruses and to continually assess future changes that may increase their pandemic potential.

    IMPORTANCEThe H5Nx viruses emerging in North America, Europe, and Asia pose a great public health concern. Here we report a molecular and structural study of the major surface proteins of several H5Nx influenza viruses. Our results improve the understanding of these new viruses and provide important information on their receptor preferences and susceptibilities to antivirals, which are central to pandemic risk assessment.

  7. Identification of a novel B-cell epitope specific for avian leukosis virus subgroup J gp85 protein.

    Science.gov (United States)

    Li, Xiaofei; Zhu, Haibo; Wang, Qi; Sun, Jiashan; Gao, Yanni; Qi, Xiaole; Wang, Yongqiang; Gao, Honglei; Gao, Yulong; Wang, Xiaomei

    2015-04-01

    Avian leukosis virus subgroup J (ALV-J) is an avian oncogenic retrovirus that has caused severe economic losses in China. Gp85 protein is the main envelope protein and the most variable structural protein of ALV-J. It is also involved in virus neutralization. In this study, a specific monoclonal antibody, 4A3, was produced against the ALV-J gp85 protein. Immunofluorescence assays showed that 4A3 could react with different strains of ALV-J, including the British prototype isolate HPRS103, the American strains, an early Chinese broiler isolate, and layer isolates. A linear epitope on the gp85 protein was identified using a series of partially overlapping fragments spanning the gp85-encoding gene and subjecting them to western blot analysis. The results indicated that (134)AEAELRDFI(142) was the minimal linear epitope that could be recognized by mAb 4A3. Enzyme-linked immunosorbent assay (ELISA) revealed that chicken anti-ALV-J sera and mouse anti-ALV-J gp85 sera could also recognize the minimal linear epitope. Alignment analysis of amino acid sequences indicated that the epitope was highly conserved among 34 ALV-J strains. Furthermore, the epitope was not conserved among subgroup A and B of avian leukosis virus (ALV). Taken together, the mAb and the identified epitope may provide valuable tools for the development of new diagnostic methods for ALV-J.

  8. Identification of an antigenic domain in the N-terminal region of avian hepatitis E virus (HEV) capsid protein that is not common to swine and human HEVs.

    Science.gov (United States)

    Wang, Lizhen; Sun, Yani; Du, Taofeng; Wang, Chengbao; Xiao, Shuqi; Mu, Yang; Zhang, Gaiping; Liu, Lihong; Widén, Frederik; Hsu, Walter H; Zhao, Qin; Zhou, En-Min

    2014-12-01

    The antigenic domains located in the C-terminal 268 amino acid residues of avian hepatitis E virus (HEV) capsid protein have been characterized. This region shares common epitopes with swine and human HEVs. However, epitopes in the N-terminal 338 amino acid residues have never been reported. In this study, an antigenic domain located between amino acids 23 and 85 was identified by indirect ELISA using the truncated recombinant capsid proteins as coating antigens and anti-avian HEV chicken sera as primary antibodies. In addition, this domain did not react with anti-swine and human HEV sera. These results indicated that the N-terminal 338 amino acid residues of avian HEV capsid protein do not share common epitopes with swine and human HEVs. This finding is important for our understanding of the antigenicity of the avian HEV capsid protein. Furthermore, it has important implications in the selection of viral antigens for serological diagnosis.

  9. The role of Matrix Gla Protein in ossification and recovery of the avian growth plate

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    Harel eDan

    2012-07-01

    Full Text Available ECM mineralization is an essential physiologic process in bone, teeth, and hypertrophic cartilage. Matrix Gla Protein (MGP, an inhibitor of mineralization, is expressed by chondrocytes and vascular smooth muscle cells to inhibit calcification of those soft tissues.Tibial Dyschondroplasia (TD, a skeletal abnormality apparent as a plug of non-vascularized, non-mineralized, white opaque cartilage in the tibial growth plate of avian species can serve as a good model for studying process and genes involved in matrix mineralization and calcification. In this work, we studied the involvement of MGP in the development of TD, as well as in the processes of spontaneous and induced recovery from this syndrome. First, we found that during normal bone development, MGP is expressed in specific time and locations, starting from wide spread expression in the yet un-ossified diaphysis during embryonic development, to specific expression in hypertrophic chondrocytes adjacent to the chondro-osseous junction and the secondary ossification center just prior to calcification. In addition, we show that MGP is not expressed in the impaired TD lesion, however when the lesion begins to heal, it strongly express MGP prior to its calcification. Moreover, we show that when calcification is inhibited, a gap is formed between the expression zones of MGP and BMP2 and that this gap is closed during the healing process. To conclude, we suggest that MGP, directly or through interaction with BMP2, plays a role as ossification regulator, rather then simple inhibitor that acts prior to ossification.

  10. Avian cardiology.

    Science.gov (United States)

    Strunk, Anneliese; Wilson, G Heather

    2003-01-01

    The field of avian cardiology is continually expanding. Although a great deal of the current knowledge base has been derived from poultry data, research and clinical reports involving companion avian species have been published. This article will present avian cardiovascular anatomy and physiology, history and physical examination considerations in the avian cardiac disease patient, specific diagnostic tools, cardiovascular disease processes, and current therapeutic modalities.

  11. Comprehensive mapping of common immunodominant epitopes in the West Nile virus nonstructural protein 1 recognized by avian antibody responses.

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    Encheng Sun

    Full Text Available West Nile virus (WNV is a mosquito-borne flavivirus that primarily infects birds but occasionally infects humans and horses. Certain species of birds, including crows, house sparrows, geese, blue jays and ravens, are considered highly susceptible hosts to WNV. The nonstructural protein 1 (NS1 of WNV can elicit protective immune responses, including NS1-reactive antibodies, during infection of animals. The antigenicity of NS1 suggests that NS1-reactive antibodies could provide a basis for serological diagnostic reagents. To further define serological reagents for diagnostic use, the antigenic sites in NS1 that are targeted by host immune responses need to be identified and the potential diagnostic value of individual antigenic sites also needs to be defined. The present study describes comprehensive mapping of common immunodominant linear B-cell epitopes in the WNV NS1 using avian WNV NS1 antisera. We screened antisera from chickens, ducks and geese immunized with purified NS1 for reactivity against 35 partially overlapping peptides covering the entire WNV NS1. This study identified twelve, nine and six peptide epitopes recognized by chicken, duck and goose antibody responses, respectively. Three epitopes (NS1-3, 14 and 24 were recognized by antibodies elicited by immunization in all three avian species tested. We also found that NS1-3 and 24 were WNV-specific epitopes, whereas the NS1-14 epitope was conserved among the Japanese encephalitis virus (JEV serocomplex viruses based on the reactivity of avian WNV NS1 antisera against polypeptides derived from the NS1 sequences of viruses of the JEV serocomplex. Further analysis showed that the three common polypeptide epitopes were not recognized by antibodies in Avian Influenza Virus (AIV, Newcastle Disease Virus (NDV, Duck Plague Virus (DPV and Goose Parvovirus (GPV antisera. The knowledge and reagents generated in this study have potential applications in differential diagnostic approaches and

  12. Highly pathogenic avian influenza virus nucleoprotein interacts with TREX complex adaptor protein Aly/REF.

    Science.gov (United States)

    Balasubramaniam, Vinod R M T; Hong Wai, Tham; Ario Tejo, Bimo; Omar, Abdul Rahman; Syed Hassan, Sharifah

    2013-01-01

    We constructed a novel chicken (Gallus gallus) lung cDNA library fused inside yeast acting domain vector (pGADT7). Using yeast two-hybrid screening with highly pathogenic avian influenza (HPAI) nucleoprotein (NP) from the strain (A/chicken/Malaysia/5858/2004(H5N1)) as bait, and the Gallus gallus lung cDNA library as prey, a novel interaction between the Gallus gallus cellular RNA export adaptor protein Aly/REF and the viral NP was identified. This interaction was confirmed and validated with mammalian two hybrid studies and co-immunoprecipitation assay. Cellular localization studies using confocal microscopy showed that NP and Aly/REF co-localize primarily in the nucleus. Further investigations by mammalian two hybrid studies into the binding of NP of other subtypes of influenza virus such as the swine A/New Jersey/1976/H1N1 and pandemic A/Malaysia/854/2009(H1N1) to human Aly/REF, also showed that the NP of these viruses interacts with human Aly/REF. Our findings are also supported by docking studies which showed tight and favorable binding between H5N1 NP and human Aly/REF, using crystal structures from Protein Data Bank. siRNA knockdown of Aly/REF had little effect on the export of HPAI NP and other viral RNA as it showed no significant reduction in virus titer. However, UAP56, another component of the TREX complex, which recruits Aly/REF to mRNA was found to interact even better with H5N1 NP through molecular docking studies. Both these proteins also co-localizes in the nucleus at early infection similar to Aly/REF. Intriguingly, knockdown of UAP56 in A549 infected cells shows significant reduction in viral titer (close to 10 fold reduction). Conclusively, our study have opened new avenues for research of other cellular RNA export adaptors crucial in aiding viral RNA export such as the SRSF3, 9G8 and ASF/SF2 that may play role in influenza virus RNA nucleocytoplasmic transport.

  13. Molecular genetics of avian proteins. XIII. Protein polymorphism in three species of Australian passerines.

    Science.gov (United States)

    Manwell, C; Baker, C M

    1975-12-01

    An introduced species, the house sparrow (Passer domesticus), and two Australian native species, the welcome swallow (Hirundo tahitica neoxena) and the fairy martin (Petrochelidon ariel), have moderately low levels of protein polymorphism compared with domesticated or semi-wild 'managed' species of birds. Genetically varient proteins in these birds include transferrin, esterase, phosphoglucomutase, NADP-dependent isocitrate dehydrogenases, phosphogluconate dehydrogenase (decarboxylating) and glucose-6-phosphate dehydrogenase. Egg-white protein polymorphism confirms heterogeneity of egg colour, markings and shape, and suggests that approximately 10% of the 'clutches' in house sparrow nests represent infidelity (intraspecific nest parasitism). For the four enzymes capable of supplying reduced NADP for reductive biosyntheses in growth and detoxification, the house sparrow has more heterozygosity (29%) than either the welcome swallow (9-4%) or the fairy martin (2-3%) and the difference is highly significant statistically. The results are discussed in relation to possible biochemical correlates of MacArthur and Wilson's (1967) evolutionary strategies or r or K selection.

  14. Structural Basis for the Development of Avian Virus Capsids That Display Influenza Virus Proteins and Induce Protective Immunity

    OpenAIRE

    Pascual, Elena; Mata, Carlos P.; Gómez-Blanco, Josué; Moreno, Noelia; Bárcena, Juan; Blanco, Esther; Rodríguez-Frandsen, Ariel; Nieto, Amelia; Carrascosa, José L.; Castón, José R.

    2014-01-01

    Bioengineering of viruses and virus-like particles (VLPs) is a well-established approach in the development of new and improved vaccines against viral and bacterial pathogens. We report here that the capsid of a major avian pathogen, infectious bursal disease virus (IBDV), can accommodate heterologous proteins to induce protective immunity. The structural units of the ∼70-nm-diameter T=13 IBDV capsid are trimers of VP2, which is made as a precursor (pVP2). The pVP2 C-terminal domain has an am...

  15. A Cistanches Herba Fraction/β-Sitosterol Causes a Redox-Sensitive Induction of Mitochondrial Uncoupling and Activation of Adenosine Monophosphate-Dependent Protein Kinase/Peroxisome Proliferator-Activated Receptor γ Coactivator-1 in C2C12 Myotubes: A Possible Mechanism Underlying the Weight Reduction Effect

    Directory of Open Access Journals (Sweden)

    Hoi Shan Wong

    2015-01-01

    Full Text Available Previous studies have demonstrated that HCF1, a semipurified fraction of Cistanches Herba, causes weight reduction in normal diet- and high fat diet-fed mice. The weight reduction was associated with the induction of mitochondrial uncoupling and changes in metabolic enzyme activities in mouse skeletal muscle. To further investigate the biochemical mechanism underlying the HCF1-induced weight reduction, the effect of HCF1 and its active component, β-sitosterol (BSS, on C2C12 myotubes was examined. Incubation with HCF1/BSS caused a transient increase in mitochondrial membrane potential (MMP, possibly by fluidizing the mitochondrial inner membrane. The increase in MMP was paralleled to an increase in mitochondrial reactive oxygen species (ROS production. Mitochondrial ROS, in turn, triggered a redox-sensitive induction of mitochondrial uncoupling by uncoupling protein 3 (UCP3. Biochemical analysis indicated that HCF1 was capable of activating an adenosine monophosphate-dependent protein kinase/peroxisome proliferator-activated receptor γ coactivator-1 pathway and thereby increased the expression of cytochrome c oxidase and UCP3. Animal studies using mitochondrial recoupler also confirmed the role of mitochondrial uncoupling in the HCF1-induced weight reduction. In conclusion, a HCF1/BSS causes the redox-sensitive induction of mitochondrial uncoupling and activation of AMPK/PGC-1 in C2C12 myotubes, with resultant reductions in body weight and adiposity by increased energy consumption.

  16. Forkhead box protein A1 inhibits the expression of uncoupling protein 2 in hydrogen peroxide-induced A549 cell line.

    Science.gov (United States)

    Song, Lan; Xu, Zhaojun; Li, Ling; Hu, Mei; Cheng, Lijuan; Chen, Lingli; Zhang, Bo

    2014-01-01

    Forkhead box protein A1 (FoxA1) is a transcription factor that is involved in embryonic development and cell differentiation. In this study, we show that hydrogen peroxide (H2O2) treatment upregulated expression of FoxA1 and UCP2 in the A549 cell line. Overexpression of FoxA1 by full-length complementary DNA reduced UCP2 expression, while silencing of FoxA1 expression by small interfering RNA significantly increased UCP2 levels. FoxA1 binds to a site from -919 to -913 bp relative to the UCP2 transcription start site. The overexpression of FoxA1 promoted the DNA binding activity and attenuated the transcription of UCP2 promoter as shown by electromobility shift, chromatin immunoprecipitation assays, and luciferase reporter assay. These data indicate an important role of FoxA1 in regulating expression of UCP2.

  17. 解偶联蛋白在脑缺血疾病中的作用研究进展%Progress in research of uncoupling protein in cerebral ischemia

    Institute of Scientific and Technical Information of China (English)

    吴传鸿; 李韶菁; 杨洪军; 陈畅; 张宁; 李德凤

    2012-01-01

    解偶联蛋白( uncoupling proteins,UCPs)是一类位于线粒体内膜上的阴离子载体蛋白,属于线粒体载体蛋白超家族.到目前为止,已经发现并确认UCPs同系物为5种,即UCP1~UCP5,中枢神经系统中主要含有UCP2,UCP4和UCP5 3种亚型,已经证实其对于脑缺血后的氧化应激、线粒体膜电势及ATP绝对水平的维持具有重要的作用,本文主要将UCPs在脑缺血疾病中相关作用进行综述,希望可以为脑缺血疾病的治疗提供新的研究思路.%The uncoupling proteins (UCPs) are a superfamily of mitochondrial anion-carrier proteins that are located on the inner mitochondrial membrane. So far, there are five homologues of UCPs identified. They are UCP1 to UCP5. The UCPs in the central nervous system is mainly composed of three subtypes, UCP2, UCP4 and UCPS. It has been confirmed that they relate to oxidative stress, mitochondrial membrane potential and absolute ATP levels after cerebral ischemia. UCPs provide a new strategy for the treatment of cerebral ischemia. In this paper, we reviewed the physiological function of UCPs and the changes after cerebral ischemia.

  18. Preparation and immune activity analysis of H5N1 subtype avian influenza virus recombinant protein-based vaccine.

    Science.gov (United States)

    Xie, Q M; Ji, J; Du, L Q; Cao, Y C; Wei, L; Xue, C Y; Qin, J P; Ma, J Y; Bi, Y Z

    2009-08-01

    Avian influenza is a severe disease among farmed poultry and free-living birds and a constant threat to the commercial chicken industry around the world. Hemagglutinin (HA) is the major immunogen on the envelope of influenza A virus and is the predominant inducer of neutralizing antibody. To obtain the bioactive antigen proteins in large quantities, a new protein expression vector pBCX was constructed, which is based on the pET32a vector. The HA gene of the H5N1 subtype of avian influenza virus (AIV) was inserted into the pBCX vector and expressed efficiently in Escherichia coli BL21 (DE3). Fused expression of the exogenous gene and msyB produced a 97-kDa msyB-HA fusion protein. Sodium dodecyl sulfate-PAGE combined with scanning analysis demonstrated that the msyB-HA fusion protein accounted for 29.5% of the total bacterial protein, 90.5% being soluble. The msyB-HA fusion protein was purified with nondenaturing 50% Ni-NTA column chromatography, and the result showed that 24 mg of purified msyB-HA fusion protein could be obtained from 1 L of induced expression bacterial culture medium. The comparative results in the present study showed that pBCX was superior to pET32a as a protein expression vector. Western blotting showed the recombinant msyB-HA (rHA) to have better antigenic activity, which may be the result from the better posttranslation protein modification and folding in the pBCX expression system. With the rHA fusion protein as antigen, we successfully prepared and screened specific monoclonal antibodys against the H5N1 subtype AIV, which indicated that the rHA had antigen epitopes and biofunctions. The immune test confirmed that the rHA protein vaccine could also induce high neutralizing antibodies, and the AIV challenge test proved that the rHA protein-based vaccine could prevent the corresponding infection. This study demonstrates that the recombinant HA protein produced by the pBCX expression system could be used as a recombinant protein-based vaccine

  19. Diversity, physiology, and evolution of avian plumage carotenoids and the role of carotenoid-protein interactions in plumage color appearance.

    Science.gov (United States)

    LaFountain, Amy M; Prum, Richard O; Frank, Harry A

    2015-04-15

    The diversity of vibrant plumage colors in birds has evolved as a direct result of social and environmental pressures. To fully understand these underlying pressures it is necessary to elucidate the mechanisms for the creation of novel plumage colors which include the metabolic transformations of dietary carotenoids and spectral tuning of the molecules within the feather protein environment. Recent advances in this field have greatly expanded the number and breadth of avian species for which plumage pigmentation has been characterized, making it possible to reconstruct the phylogenetic history of carotenoid usage in plumage. Resonance Raman and classical Raman spectroscopic techniques have been employed with great effect in the study of carotenoids in situ. The application of these methods have two benefits: to identify carotenoids in feathers that are unavailable for destructive sampling; and to study the spectral tuning resulting from the interaction between the carotenoids and the proteins to which they are bound. This review presents a summary of recent advances in the understanding of the molecular factors controlling the coloration of avian carotenoid plumage obtained through the application of both bioanalytical and spectroscopic methodologies.

  20. Avian anemia's

    Directory of Open Access Journals (Sweden)

    Raukar Jelena

    2005-01-01

    Full Text Available This paper deals with avian anemia's classified by MCHC/MCV and with types of anemia's. Father hematological and immunological research is needed to secure information on hematological parameters in different avian species at their earliest age. Anemia is a common clinical finding in birds because the avian erythrocyte half - life is much shorter than the mammalian. Therefore anemia should be determined as soon as possible. Researchers should standardize hematological parameters for every single avian species.

  1. Brown adipose tissue mitochondria oxidizing fatty acids generate high levels of reactive oxygen species irrespective of the uncoupling protein-1 activity state.

    Science.gov (United States)

    Schönfeld, Peter; Wojtczak, Lech

    2012-03-01

    Mitochondria from brown adipose tissue (BATM) have a high enzymatic capacity for fatty acid oxidation and therefore are an ideal model to examine the sites of reactive oxygen species (ROS) generation during fatty acid oxidation. ROS generation by BATM (isolated from 3-week-old rats) was measured during acylcarnitine oxidation as release of H(2)O(2) into the medium and as inactivation of the matrix enzyme aconitase. The following results were obtained: (1) BATM release large amounts of H(2)O(2) in the coupled as well as in the uncoupled states, several times more than skeletal muscle mitochondria. (2) H(2)O(2) release is especially large with acylcarnitines of medium-chain fatty acids (e.g. octanoylcarnitine). (3) Reverse electron transport does not contribute in a significant extent to the overall ROS generation. (4) Despite the large release of H(2)O(2), the ROS-sensitive matrix enzyme aconitase is not inactivated during acylcarnitine oxidation. (5) In contrast to acylcarnitines, oxidation of α-glycerophosphate by BATM is characterized by large H(2)O(2) release and a pronounced aconitase inactivation. We hypothesize that acylcarnitine-supported ROS generation in BATM may be mainly associated with acyl-CoA dehydrogenase and electron transferring flavoprotein-ubiquinone reductase rather than with complexes of the respiratory chain.

  2. Effects of Constant Moderate Temperatures on Performance, Glucose and Lipid Metabolism, Expression of Uncoupling Protein of broilers%持续偏热环境对肉鸡生长性能、糖脂代谢及解偶联蛋白 mRNA表达的影响

    Institute of Scientific and Technical Information of China (English)

    甄龙; 石玉祥; 张敏红; 冯京海; 张少帅; 彭骞骞

    2015-01-01

    本试验研究了持续不同温度处理( 21、26和31 ℃)对肉鸡生长性能、血清糖脂代谢相关指标、胸肌和肝脏解偶联蛋白( avUCP) mRNA表达的影响. 试验选取22日龄爱拔益加( AA)肉鸡144只转入环境控制舱,随机分成3组,每组6个重复,每个重复8只鸡(公母各4只). 适应期7 d,温度21℃,相对湿度60%. 29日龄时,试验温度分别调整到21、26和31℃,相对湿度60%,直至试验结束,共14 d. 结果表明:1 ) 31 ℃组肉鸡平均日增重( ADG )、平均日采食量( ADFI)极显著低于21、26 ℃组( P0.05). 2) 31 ℃组肉鸡血清葡萄糖( GLU)、总胆固醇( TC)、甘油三酯( TG)、游离脂肪酸( FFA)含量显著高于21℃组(P0.05). 3)31℃组肉鸡血清甲状腺素(T4)、瘦素(LEP)、皮质酮(CORT)含量显著高于21 ℃组(P0.05). 4)试验第7天,31 ℃组胸肌avUCP mRNA表达显著低于21、26 ℃组( P0.05) in F/G was found between the two groups. 2) serum glucose ( GLU) , total cholesterol ( TC) , total triglyceride ( TG) and nonestesterified fatty acid ( FFA) in 31℃ were significant ( P0.05) was found between 21 and 26 ℃ groups. 3) serum thyroxine (T4), leptin (LEP) and corticoster-one ( CORT) in 31 ℃ were significant ( P0.05) was found between 21 and 26℃ groups except for T4. 4) expression of avian uncoupling pro-tein (avUCP) mRNA in pectoral muscle of broilers in 31 ℃ group showed highly significant (P<0.01) de-pression than that in 21 and 26 ℃ groups at the seventh day, with highly significant ( P<0.01) depression in 26 and 31℃ groups than 21℃ at the fourteenth day;throughout the trial period, expression of avUCP mRNA in liver of broilers in 31 ℃ group was highly significant ( P<0.01) higher than that in 21 and 26 ℃ groups. In conclusion, compared with 21 ℃ group, the constant moderate heat stress ( 26 and 31 ℃) can the perform-ance, glucose and lipid metabolism, and expression of uncoupling protein of broilers. And the different extent moderate heat

  3. Sequence and epitope analysis of surface proteins of avian influenza H5N1 viruses from Asian patients

    Institute of Scientific and Technical Information of China (English)

    LI Guanglin; TAO Shiheng; WANG Xiujie

    2006-01-01

    Increasing cases of human infections with the high pathogenic avian influenza virus H5N1 have raised great concern on potential human flu pandemics caused by H5N1. The two viral surface glycoproteins, the hemagglutinin (HA) and the neuraminidase (NA) proteins, are major antigens of H5N1. Introducing new mutations on these two proteins is the major strategy used by H5N1 to expand host range and to avoid the recognition of host immune systems. We analyzed the two surface proteins of H5N1 from Asian human patients and identified many new mutation sites, including a few that were unique to certain lethal strains. We also analyzed the distribution of mutations on different epitopes of the two surface proteins. A receptor-binding site that might involve in the determination of host specificity of H5N1 was also found. Results reported here provided information for better understanding of the evolution trend of H5N1 genome in human.

  4. Antigenic cross-reactivity among avian pneumoviruses of subgroups A, B, and C at the matrix but not nucleocapsid proteins.

    Science.gov (United States)

    Lwamba, Humphrey C M; Halvorson, David A; Nagaraja, Kakambi V; Turpin, Elizabeth A; Swayne, David; Seal, Bruce S; Njenga, M Kariuki

    2002-01-01

    Earlier findings from our laboratory based on analysis of nucleotide and predicted amino acid sequence identities of 15 avian pneumoviruses (APVs) isolated from the United States (subgroup C) demonstrated that the viruses were phylogenetically separated from the European subgroup A and subgroup B viruses. Here, we investigated whether viruses from the three subgroups were cross-reactive by testing field sera positive for each of the APV subgroups in an enzyme-linked immunosorbent assay (ELISA) test with recombinant matrix (M) and nucleoprotein (N) proteins generated from a Minnesota APV isolate (APV/MN2A). Sera from turkeys infected with APV subgroup A, B, or C reacted with recombinant M protein derived from APV/MN2A. In contrast, recombinant N protein from APV/MN2A virus was reactive with sera from subtypes A and C viruses but not from subtype B virus. The results illustrate that viruses from the three APV subtypes share antigenic homology, and the M protein-based ELISA is adequate for monitoring APV outbreaks but not for distinguishing between different subtypes.

  5. Genetic Variance in Uncoupling Protein 2 in Relation to Obesity, Type 2 Diabetes, and Related Metabolic Traits: Focus on the Functional −866G>A Promoter Variant (rs659366

    Directory of Open Access Journals (Sweden)

    Louise T. Dalgaard

    2011-01-01

    Full Text Available Uncoupling proteins (UCPs are mitochondrial proteins able to dissipate the proton gradient of the inner mitochondrial membrane when activated. This decreases ATP-generation through oxidation of fuels and may theoretically decrease energy expenditure leading to obesity. Evidence from Ucp(−/− mice revealed a role of UCP2 in the pancreatic β-cell, because β-cells without UCP2 had increased glucose-stimulated insulin secretion. Thus, from being a candidate gene for obesity UCP2 became a valid candidate gene for type 2 diabetes mellitus. This prompted a series of studies of the human UCP2 and UCP3 genes with respect to obesity and diabetes. Of special interest was a promoter variant of UCP2 situated 866bp upstream of transcription initiation (−866G>A, rs659366. This variant changes promoter activity and has been associated with obesity and/or type 2 diabetes in several, although not all, studies. The aim of the current paper is to summarize current evidence of association of UCP2 genetic variation with obesity and type 2 diabetes, with focus on the −866G>A polymorphism.

  6. Screening of Proteins Interacting with Nonstructural 1 Protein of H5N1 Avian Influenza Virus from T7-phage Display Library

    Institute of Scientific and Technical Information of China (English)

    ZHU Chun-yu; WU Jian-guo; LIU Hong-sheng; SUN Ting-ting; ZHAO Jian; WANG Ning; ZHENG Fang-liang; AI Hai-xin; ZHU Jun-feng; WANG Xiao-ying; ZHU Ying

    2012-01-01

    Avian influenza virus(AIV) nonstructural 1(NS1) gene was amplified by real-time polymerse chain reaction(RT-PCR) and inserted into pET28a,then transformed into E.coli BL21(DE3) competent cell.With the induction of isopropyl-β-D-thiogalactoside(IPTG) and the purification of Ni-NTA column,we finally obtained purified NS1 protein.T7-phage display system was used to screen the proteins that interacted with NS1 from lung cell cDNA library.The selected positive clones were identified by DNA sequencing and analyzed by BLAST program in GeneBank.Two proteins were obtained as NS 1 binding proteins,Homo sapiens nucleolar and coiled-body phosphoprotein 1(NOLC1) and Homo sapiens similar to colon cancer-associated antigen.By co-immunoprecipitation and other methods,Homo sapiens NOLC1 was found to interact with the NS1 protein,the results would provide the basis for further studying biological function of NS1 protein.

  7. Sequence analysis of the large polymerase (L) protein of the US strain of avian metapneumovirus indicates a close resemblance to that of the human metapneumovirus.

    Science.gov (United States)

    Govindarajan, Dhanasekaran; Samal, Siba K

    2004-09-15

    The complete nucleotide sequence of the large polymerase (L) protein of the avian metapneumovirus subgroup C strain Colorado was determined. The L protein gene of avian pneumovirus Colorado isolate (APV-C) was 6173 nucleotides in length from the gene-start to the gene-end and encoded a polypeptide of 2005 amino acids in length. The length of the L protein of APV-C was exactly the same as that of human metapneumovirus (hMPV) and one amino acid longer than the L protein of APV subgroup A. The L protein of APV-C showed 80% amino acid identity with the L protein of hMPV, but only 64% amino acid identity with the L protein of APV-A. The nucleotide and deduced amino acid sequences were compared with the corresponding sequences of eleven other paramyxoviruses. All six domains characteristic of paramyxovirus L proteins were also observed in the L protein of APV-C. All the polymerase core motifs in domain III were conserved to nearly 100% in the metapneumoviruses. Similarly, the putative ATP-binding motif in domain VI was completely conserved among the metapneumoviruses and differed in length, by one intermediate residue, from other paramyxoviruses. Phylogenetic analysis of the different L proteins also revealed a closer relationship between APV-C and hMPV.

  8. Cell-specific targeting of lentiviral vectors mediated by fusion proteins derived from Sindbis virus, vesicular stomatitis virus, or avian sarcoma/leukosis virus

    Directory of Open Access Journals (Sweden)

    Marino Michael P

    2010-01-01

    Full Text Available Abstract Background The ability to efficiently and selectively target gene delivery vectors to specific cell types in vitro and in vivo remains one of the formidable challenges in gene therapy. We pursued two different strategies to target lentiviral vector delivery to specific cell types. In one of the strategies, vector particles bearing a membrane-bound stem cell factor sequence plus a separate fusion protein based either on Sindbis virus strain TR339 glycoproteins or the vesicular stomatitis virus G glycoprotein were used to selectively transduce cells expressing the corresponding stem cell factor receptor (c-kit. An alternative approach involved soluble avian sarcoma/leukosis virus receptors fused to cell-specific ligands including stem cell factor and erythropoietin for targeting lentiviral vectors pseudotyped with avian sarcoma/leukosis virus envelope proteins to cells that express the corresponding receptors. Results The titers of unconcentrated vector particles bearing Sindbis virus strain TR339 or vesicular stomatitis virus G fusion proteins plus stem cell factor in the context of c-kit expressing cells were up to 3.2 × 105 transducing units per ml while vector particles lacking the stem cell factor ligand displayed titers that were approximately 80 fold lower. On cells that lacked the c-kit receptor, the titers of stem cell factor-containing vectors were approximately 40 times lower compared to c-kit-expressing cells. Lentiviral vectors pseudotyped with avian sarcoma/leukosis virus subgroup A or B envelope proteins and bearing bi-functional bridge proteins encoding erythropoietin or stem cell factor fused to the soluble extracellular domains of the avian sarcoma/leukosis virus subgroup A or B receptors resulted in efficient transduction of erythropoietin receptor or c-kit-expressing cells. Transduction of erythropoietin receptor-expressing cells mediated by bi-functional bridge proteins was found to be dependent on the dose, the

  9. Accute satiety response of mammalian, avian and fish proteins in dogs

    Science.gov (United States)

    Fish proteins have been reported to be more satiating than meat proteins. The objective was to determine the effect of different animal protein pre-meals on satiety. Ten intact female hounds were fed either pork loin, beef loin, chicken breast, salmon fillet, or pollock fillet. During Phase I, 100 g...

  10. Topology and cellular localization of the small hydrophobic protein of avian metapneumovirus

    Science.gov (United States)

    The small hydrophobic protein (SH) is a type II integral membrane protein that is packaged into virions and is only present in certain paramyxoviruses including metapneumovirus. In addition to a highly divergent primary sequence, SH proteins vary significantly in size among the different viruses. Hu...

  11. Uncoupling of Obesity from Insulin Resistance Through a Targeted Mutation in aP2, the Adipocyte Fatty Acid Binding Protein

    Science.gov (United States)

    Hotamisligil, Gokhan S.; Johnson, Randall S.; Distel, Robert J.; Ellis, Ramsey; Papaioannou, Virginia E.; Spiegelman, Bruce M.

    1996-11-01

    Fatty acid binding proteins (FABPs) are small cytoplasmic proteins that are expressed in a highly tissue-specific manner and bind to fatty acids such as oleic and retinoic acid. Mice with a null mutation in aP2, the gene encoding the adipocyte FABP, were developmentally and metabolically normal. The aP2-deficient mice developed dietary obesity but, unlike control mice, they did not develop insulin resistance or diabetes. Also unlike their obese wild-type counterparts, obese aP2-/- animals failed to express in adipose tissue tumor necrosis factor-α (TNF-α), a molecule implicated in obesity-related insulin resistance. These results indicate that aP2 is central to the pathway that links obesity to insulin resistance, possibly by linking fatty acid metabolism to expression of TNF-α.

  12. Avian Wings

    Science.gov (United States)

    Liu, Tianshu; Kuykendoll, K.; Rhew, R.; Jones, S.

    2004-01-01

    This paper describes the avian wing geometry (Seagull, Merganser, Teal and Owl) extracted from non-contact surface measurements using a three-dimensional laser scanner. The geometric quantities, including the camber line and thickness distribution of airfoil, wing planform, chord distribution, and twist distribution, are given in convenient analytical expressions. Thus, the avian wing surfaces can be generated and the wing kinematics can be simulated. The aerodynamic characteristics of avian airfoils in steady inviscid flows are briefly discussed. The avian wing kinematics is recovered from videos of three level-flying birds (Crane, Seagull and Goose) based on a two-jointed arm model. A flapping seagull wing in the 3D physical space is re-constructed from the extracted wing geometry and kinematics.

  13. Avian influenza

    Science.gov (United States)

    ... of avian influenza A in Asia, Africa, Europe, Indonesia, Vietnam, the Pacific, and the near East. Hundreds ... to detect abnormal breath sounds) Chest x-ray Culture from the nose or throat A method or ...

  14. Avian Influenza

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This is a letter from a professor at Clemson University about waterfowl that had been tested for avian influenza at Santee National Wildlife Refuge

  15. Avian hematology.

    Science.gov (United States)

    Jones, Michael P

    2015-01-01

    Avian veterinarians often rely heavily on the results of various diagnostic tests, including hematology results. As such, cellular identification and evaluation of the cellular response are invaluable tools that help veterinarians understand the health or condition of their patient, as well as to monitor severity and clinical progression of disease and response to treatment. Therefore, it is important to thoroughly understand how to identify and evaluate changes in the avian erythron and leukon, as well as to interpret normal and abnormal results.

  16. Avian Flu

    Energy Technology Data Exchange (ETDEWEB)

    Eckburg, Paul

    2006-11-06

    Since 2003, a severe form of H5N1 avian influenza has rapidly spread throughout Asia and Europe, infecting over 200 humans in 10 countries. The spread of H5N1 virus from person-to-person has been rare, thus preventing the emergence of a widespread pandemic. However, this ongoing epidemic continues to pose an important public health threat. Avian flu and its pandemic potential in humans will be discussed.

  17. Avian Research

    Institute of Scientific and Technical Information of China (English)

    2016-01-01

    Aims and Scope Avian Research is an open access,peer-reviewed journal publishing high quality research and review articles on all aspects of ornithology from all over the world.It aims to report the latest and most significant progress in ornithology and to encourage exchange of ideas among international ornithologists.As an Open Access journal,Avian Research provides a unique opportunity to publish

  18. Avian Research

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    <正>Aims and Scope Avian Research is an open access,peer-reviewed journal publishing high quality research and review articles on all aspects of ornithology from all over the world.It aims to report the latest and most significant progress in ornithology and to encourage exchange of ideas among international ornithologists.As an Open Access journal,Avian Research provides a unique opportunity to publish high quality contents that will be internationally accessible to any reader at no cost.

  19. Avian Research

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    Aims and Scope Avian Research is an open access,peer-reviewed journal publishing high quality research and review articles on all aspects of ornithology from all over the world.It aims to report the latest and most signi cant progress in ornithology and to encourage exchange of ideas among international ornithologists.As an Open Access journal,Avian Research provides a unique opportunity to publish

  20. Evaluation of the refractometric method for the determination of total protein in avian plasma or serum.

    Science.gov (United States)

    Lumeij, J T; de Bruijne, J J

    1985-07-01

    Serum total protein concentrations in pigeon blood determined with the biuret method (TPB-se) were compared with total protein concentrations in plasma (TPR-pl) and serum (TPR-se) obtained by estimation from refractive index. The refractometric method consistently yielded higher values (Prefractometric method for determination of TP in pigeon blood is not recommended.

  1. Protein-matrix coupling/uncoupling in "dry" systems of photosynthetic reaction center embedded in trehalose/sucrose: the origin of trehalose peculiarity.

    Science.gov (United States)

    Francia, Francesco; Dezi, Manuela; Mallardi, Antonia; Palazzo, Gerardo; Cordone, Lorenzo; Venturoli, Giovanni

    2008-08-06

    Trehalose is a nonreducing disaccharide of glucose found in organisms, which can survive adverse conditions such as extreme drought and high temperatures. Furthermore, isolated structures, as enzymes or liposomes, embedded in trehalose are preserved against stressing conditions [see, e.g., Crowe, L. M. Comp. Biochem. Physiol. A 2002, 131, 505-513]. Among other hypotheses, such protective effect has been suggested to stem, in the case of proteins, from the formation of a water-mediated, hydrogen bond network, which anchors the protein surface to the water-sugar matrix, thus coupling the internal degrees of freedom of the biomolecule to those of the surroundings [Giuffrida, S.; et al. J. Phys. Chem. B 2003, 107, 13211-13217]. Analogous protective effect is also accomplished by other saccharides, although with a lower efficiency. Here, we studied the recombination kinetics of the primary, light-induced charge separated state (P(+)Q(A)(-)) and the thermal stability of the photosynthetic reaction center (RC) of Rhodobacter sphaeroides in trehalose-water and in sucrose-water matrixes of decreasing water content. Our data show that, in sucrose, at variance with trehalose, the system undergoes a "nanophase separation" when the water/sugar mole fraction is lower than the threshold level approximately 0.8. We rationalize this result assuming that the hydrogen bond network, which anchors the RC surface to its surrounding, is formed in trehalose but not in sucrose. We suggest that both the couplings, in the case of trehalose, and the nanophase separation, in the case of sucrose, start at low water content when the components of the system enter in competition for the residual water.

  2. Expression of hemagglutinin protein from the avian influenza virus H5N1 in a baculovirus/insect cell system significantly enhanced by suspension culture

    Directory of Open Access Journals (Sweden)

    Spencer Lynn

    2006-02-01

    Full Text Available Abstract Background Prevention of a possible avian influenza pandemic necessitates the development of rapid diagnostic tests and the eventual production of a vaccine. Results For vaccine production, hemagglutinin (HA1 from avian influenza H5N1 was expressed from a recombinant baculovirus. Recombinant HA1 was expressed in monolayer or suspension culture insect cells by infection with the recombinant baculovirus. The yield of rHA1 from the suspension culture was 68 mg/l, compared to 6 mg/l from the monolayer culture. Immunization of guinea pigs with 50 μg of rHA1 yielded hemagglutinin inhibition and virus neutralization titers of 1:160 after two times vaccination with rHA1 protein. Conclusion Thus, the production of rHA1 using an insect suspension cell system provides a promising basis for economical production of a H5 antigen.

  3. 过表达解偶联蛋白-2基因加重原代肝细胞急性损伤%Overexpression of Uncoupling Protein 2 Aggravates Acute Primary Hepatocyte Damage

    Institute of Scientific and Technical Information of China (English)

    万赤丹; 程锐; 王春友; 王宏博; 王帅; 刘涛

    2009-01-01

    目的 研究解偶联蛋白-2(uncoupling protein-2,UCP-2)基因在肝细胞急性损伤过程中的作用.方法 构建小鼠UCP-2基因过表达慢病毒载体;二步胶原酶灌注法分离培养小鼠原代肝细胞;慢病毒感染肝细胞,使肝细胞中UCP-2基因表达上调;荧光显微镜镜检确定感染效率.Real time PCR检测UCP-2表达;肿瘤坏死因子-α(TNF-α)作用于病毒感染肝细胞24 h后.检测细胞上清液中谷丙转氨酶(ALT)和乳酸脱氢酶(LDH)水平;碘化丙啶(PI)染色流式细胞仪检测细胞凋亡,Western blot检测凋亡因子Caspase 3的活化.结果 Real time PCR检测证实目的 细胞中UCP-2表达上调;TNF-α作用后病毒感染的实验组上清液中ALT和LDH水平、细胞凋亡率、凋亡因子Caspase 3活化程度均高于对照组(均P<0.05).结论 UCP-2基因过度表达可加重肝细胞急性损伤.

  4. Convergent effects on cell signaling mechanisms mediate the actions of different neurobehavioral teratogens: alterations in cholinergic regulation of protein kinase C in chick and avian models.

    Science.gov (United States)

    Yanai, Joseph; Beer, Avital; Huleihel, Rabab; Izrael, Michal; Katz, Sofia; Levi, Yaarit; Rozenboim, Israel; Yaniv, Shiri P; Slotkin, Theodore A

    2004-10-01

    Although the actions of heroin on central nervous system (CNS) development are mediated through opioid receptors, the net effects converge on dysfunction of cholinergic systems. We explored the mechanisms underlying neurobehavioral deficits in mouse and avian (chick, Cayuga duck) models. In mice, prenatal heroin exposure (10 mg/kg on gestation days 9-18) elicited deficits in behaviors related to hippocampal cholinergic innervation, characterized by concomitant pre- and postsynaptic hyperactivity, but ending in a reduction of basal levels of protein kinase C (PKC) isoforms betaII and gamma and their desensitization to cholinergic receptor-induced activation. PKCalpha, which is not involved in the behaviors studied, was unaffected. Because mammalian models possess inherent confounding factors from maternal effects, we conducted parallel studies using avian embryos, evaluating hyperstriatal nucleus (intermedial part of the hyperstriatum ventrale, IMHV)-related, filial imprinting behavior. Heroin injection to the eggs (20 mg/kg) on incubation days 0 and 5 diminished the post-hatch imprinting ability and reduced PKCg and bII content in the IMHV membrane fraction. Two otherwise unrelated agents that converge on cholinergic systems, chlorpyrifos and nicotine, elicited the same spectrum of effects on PKC isoforms and imprinting but had more robust actions. Pharmacological characterization also excluded direct effects of opioid receptors on the expression of imprinting; instead, it indicated participation of serotonergic innervation. The avian models can provide rapid screening of neuroteratogens, exploration of common mechanisms of behavioral disruption, and the potential design of therapies to reverse neurobehavioral deficits.

  5. Analysis of viral protein-2 encoding gene of avian encephalomyelitis virus from field specimens in Central Java region, Indonesia

    Directory of Open Access Journals (Sweden)

    Aris Haryanto

    2016-01-01

    Full Text Available Aim: Avian encephalomyelitis (AE is a viral disease which can infect various types of poultry, especially chicken. In Indonesia, the incidence of AE infection in chicken has been reported since 2009, the AE incidence tends to increase from year to year. The objective of this study was to analyze viral protein 2 (VP-2 encoding gene of AE virus (AEV from various species of birds in field specimen by reverse transcription polymerase chain reaction (RT-PCR amplification using specific nucleotides primer for confirmation of AE diagnosis. Materials and Methods: A total of 13 AEV samples are isolated from various species of poultry which are serologically diagnosed infected by AEV from some areas in central Java, Indonesia. Research stage consists of virus samples collection from field specimens, extraction of AEV RNA, amplification of VP-2 protein encoding gene by RT-PCR, separation of RT-PCR product by agarose gel electrophoresis, DNA sequencing and data analysis. Results: Amplification products of the VP-2 encoding gene of AEV by RT-PCR methods of various types of poultry from field specimens showed a positive results on sample code 499/4/12 which generated DNA fragment in the size of 619 bp. Sensitivity test of RT-PCR amplification showed that the minimum concentration of RNA template is 127.75 ng/μl. The multiple alignments of DNA sequencing product indicated that positive sample with code 499/4/12 has 92% nucleotide homology compared with AEV with accession number AV1775/07 and 85% nucleotide homology with accession number ZCHP2/0912695 from Genbank database. Analysis of VP-2 gene sequence showed that it found 46 nucleotides difference between isolate 499/4/12 compared with accession number AV1775/07 and 93 nucleotides different with accession number ZCHP2/0912695. Conclusions: Analyses of the VP-2 encoding gene of AEV with RT-PCR method from 13 samples from field specimen generated the DNA fragment in the size of 619 bp from one sample with

  6. Avian reovirus nonstructural protein p17-induced G(2)/M cell cycle arrest and host cellular protein translation shutoff involve activation of p53-dependent pathways.

    Science.gov (United States)

    Chulu, Julius L C; Huang, Wei R; Wang, L; Shih, Wen L; Liu, Hung J

    2010-08-01

    The effects of avian reovirus (ARV) p17 protein on cell cycle progression and host cellular protein translation were studied. ARV infection and ARV p17 transfection resulted in the accumulation of infected and/or transfected cells in the G(2)/M phase of the cell cycle. The accumulation of cells in the G(2)/M phase was accompanied by upregulation and phosphorylation of the G(2)/M-phase proteins ATM, p53, p21(cip1/waf1), Cdc2, cyclin B1, Chk1, Chk2, and Cdc25C, suggesting that p17 induces a G(2)/M cell cycle arrest through activation of the ATM/p53/p21(cip1/waf1)/Cdc2/cyclin B1 and ATM/Chk1/Chk2/Cdc25C pathways. The G(2)/M cell cycle arrest resulted in increased virus replication. In the present study, we also provide evidence demonstrating that p17 protein is responsible for ARV-induced host cellular protein translation shutoff. Increased phosphorylation levels of the eukaryotic translation elongation factor 2 (eEF2) and initiation factor eIF2alpha and reduced phosphorylation levels of the eukaryotic translation initiation factors eIF4E, eIF4B, and eIF4G, as well as 4E-BP1 and Mnk-1 in p17-transfected cells, demonstrated that ARV p17 suppresses translation initiation factors and translation elongation factors to induce host cellular protein translation shutoff. Inhibition of mTOR by rapamycin resulted in a decrease in the levels of phosphorylated 4E-BP1, eIF4B, and eIF4G and an increase in the levels eEF2 but did not affect ARV replication, suggesting that ARV replication was not hindered by inhibition of cap-dependent translation. Taken together, our data indicate that ARV p17-induced G(2)/M arrest and host cellular translation shutoff resulted in increased ARV replication.

  7. Avian Research

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    Aims and Scope Avian Research is an open access,peer-reviewed journal publishing high quality research and review articles on all aspects of ornithology from all over the world.It aims to report the latest and most significant progress in ornithology and to encourage exchange of ideas among international ornithologists.As an Open Access journal,

  8. Avian Research

    Institute of Scientific and Technical Information of China (English)

    2016-01-01

    Aims and Scope Avian Research is an open access,peer-reviewed journal publishing high quality research and review articles on all aspects of ornithology from all over the world.It aims to report the latest and most significant progress in ornithology and to encourage exchange of ideas among international ornithologists.As an Open Access journal,

  9. Avian Research

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    <正>Aims and Scope Avian Research is an open access,peer-reviewed journal publishing high quality research and review articles on all aspects of ornithology from all over the world.It aims to report the latest and most significant progress in ornithology and to encourage exchange of ideas among international ornithologists.As an Open Access journal,

  10. Avian Research

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    <正>Aims and Scope Avian Research is an open access,peer-reviewed journal publishing high quality research and review articles on all aspects of ornithology from all over the world.It aims to report the latest and most significant progress in ornithology and to encourage exchange of ideas among international ornithologists.As an Open Access journal,

  11. Avian Influenza in Birds

    Science.gov (United States)

    ... this? Submit Button Past Newsletters Avian Influenza in Birds Language: English Español Recommend on Facebook Tweet ... illness. Top of Page Avian Influenza in Wild Birds Avian influenza A viruses have been isolated from ...

  12. Studies on Nanoparticle Based Avian Influenza Vaccines to Present Immunogenic Epitopes of the Virus with Concentration on Ectodomain of Matrix 2 (M2e) Protein

    Science.gov (United States)

    Babapoor Dighaleh, Sankhiros

    2011-12-01

    Avian influenza is an infectious disease of avian species caused by type A influenza viruses with a significant economic impact on the poultry industry. Vaccination is the main prevention strategy in many countries worldwide. However, available vaccines elicit antibodies against two major surface protein of the virus hemagglutinin (HA) and neuraminidase (NA), where they constantly change by point mutations. Influenza viruses can also easily undergo gene reassortment. Therefore, to protect chickens against new strain of avian influenza virus, as well as control and prevent virus spread among farms, new vaccines needed to be designed which is a tedious, time consuming and expensive. Recently, conserved regions of the influenza genome have been evaluated as possible universal vaccines to eliminate constant vaccine updates based on circulating virus. In this study, peptide nanotechnology was used to generate vaccine nanoparticles that carry the highly conserved external domain of matrix 2 protein (M2e). These nanoparticles presented M2e in monomeric or tetrameric forms, designated as PSC-M2e-CH and BNSC-M2eN-CH. respectively. First, to demonstrate immunogenicity of these nanoparticles, we measured anti-M2e antibody in chickens, particularly when a high dose was applied. Prior to vaccination-challenge study, the challenge dose were determined by oculonasal inoculation of 10 6 EID50 or 107.7 EID50 of low pathogenicity AI virus HSN2 followed by measuring cloacal and tracheal virus shedding. A biphasic virus shedding pattern was observed with two peaks of virus shedding at days 4 and 8 for both tracheal and cloacal swabs. The chickens infected with 107.7 EID50 had significant virus shedding as compared with 106 EID50. Based on results of mentioned studies, a vaccination-challenge study was conducted by using 75mug of each vaccine construct per inoculation (with and without adjuvant) and higher dose of virus for challenge. BN5C-M2e-CH with adjuvant significantly reduced the

  13. Deduced amino acid sequence of the small hydrophobic protein of US avian pneumovirus has greater identity with that of human metapneumovirus than those of non-US avian pneumoviruses.

    Science.gov (United States)

    Yunus, Abdul S; Govindarajan, Dhanasekaran; Huang, Zhuhui; Samal, Siba K

    2003-05-01

    We report here the nucleotide and deduced amino acid (aa) sequences of the small hydrophobic (SH) gene of the avian pneumovirus strain Colorado (APV/CO). The SH gene of APV/CO is 628 nucleotides in length from gene-start to gene-end. The longest ORF of the SH gene encoded a protein of 177 aas in length. Comparison of the deduced aa sequence of the SH protein of APV/CO with the corresponding published sequences of other members of genera metapneumovirus showed 28% identity with the newly discovered human metapneumovirus (hMPV), but no discernable identity with the APV subgroup A or B. Collectively, this data supports the hypothesis that: (i) APV/CO is distinct from European APV subgroups and belongs to the novel subgroup APV/C (APV/US); (ii) APV/CO is more closely related to hMPV, a mammalian metapneumovirus, than to either APV subgroup A or B. The SH gene of APV/CO was cloned using a genomic walk strategy which initiated cDNA synthesis from genomic RNA that traversed the genes in the order 3'-M-F-M2-SH-G-5', thus confirming that gene-order of APV/CO conforms in the genus Metapneumovirus. We also provide the sequences of transcription-signals and the M-F, F-M2, M2-SH and SH-G intergenic regions of APV/CO.

  14. Avian reovirus L2 genome segment sequences and predicted structure/function of the encoded RNA-dependent RNA polymerase protein

    Directory of Open Access Journals (Sweden)

    Xu Wanhong

    2008-12-01

    Full Text Available Abstract Background The orthoreoviruses are infectious agents that possess a genome comprised of 10 double-stranded RNA segments encased in two concentric protein capsids. Like virtually all RNA viruses, an RNA-dependent RNA polymerase (RdRp enzyme is required for viral propagation. RdRp sequences have been determined for the prototype mammalian orthoreoviruses and for several other closely-related reoviruses, including aquareoviruses, but have not yet been reported for any avian orthoreoviruses. Results We determined the L2 genome segment nucleotide sequences, which encode the RdRp proteins, of two different avian reoviruses, strains ARV138 and ARV176 in order to define conserved and variable regions within reovirus RdRp proteins and to better delineate structure/function of this important enzyme. The ARV138 L2 genome segment was 3829 base pairs long, whereas the ARV176 L2 segment was 3830 nucleotides long. Both segments were predicted to encode λB RdRp proteins 1259 amino acids in length. Alignments of these newly-determined ARV genome segments, and their corresponding proteins, were performed with all currently available homologous mammalian reovirus (MRV and aquareovirus (AqRV genome segment and protein sequences. There was ~55% amino acid identity between ARV λB and MRV λ3 proteins, making the RdRp protein the most highly conserved of currently known orthoreovirus proteins, and there was ~28% identity between ARV λB and homologous MRV and AqRV RdRp proteins. Predictive structure/function mapping of identical and conserved residues within the known MRV λ3 atomic structure indicated most identical amino acids and conservative substitutions were located near and within predicted catalytic domains and lining RdRp channels, whereas non-identical amino acids were generally located on the molecule's surfaces. Conclusion The ARV λB and MRV λ3 proteins showed the highest ARV:MRV identity values (~55% amongst all currently known ARV and MRV

  15. 抑制解偶联蛋白-2基因表达减轻急性脂肪肝细胞损伤%Inhibiting the expression of uncoupling protein-2 attenuates acute damage to fatty liver cells

    Institute of Scientific and Technical Information of China (English)

    程锐; 王春友; 刘涛; 王宏博; 王帅; 万赤丹

    2008-01-01

    Objective To investigate the effects of down-regulating uncoupling protein-2 (UCP-2) expression on acute damage to fatty liver cells and explore a new target for the donor liverwith steatosis. Methods Primary fatty liver cells were isolated from C57BL/6J-ob/ob transgenic miceby two-step collagenase perfusion method. RNAi lentivirus vector targeting mouse UCP-2 gene wasused to knock down the UCP-2 gene in the steatosis hepatocytes (the experimental group). Emptylentivirus vector was transfected into the steatosis hepatocytes cells as the control group. Under thefluorescence microscopy, the transfection efficiency was tested. Real time PCR was used to determinethe effect of RNAi. After the transfected cells were treated with TNF-α for 24 h, apoptosis wasanalyzed by flow cytometry using PI staining. Activation of caspase3 was detected by Western blot.Resalts The expression of UCP-2 gene was inhibited effectively, and the knockdown rate of UCP-2gene was 75%. The apoptosis rate in the experimental group was (4.97±0.25)%, significantlylower than in the control group [(21.13±1.28)%, p<0.05 ]. Activation 'of caspase3 in theexperimental group was also weaker than in the control group. Conclusion Inhibiting the expression ofUCP-2 can attenuate the injury of fatty liver cells.%目的 探讨下调解偶联蛋白-2(UCP-2)基因的表达对脂肪肝细胞损伤的影响,为存在脂肪变性的供肝提供治疗靶点.方法 采用二步胶原酶灌注法分离C57BL/6 J-ob/ob转基因肥胖小鼠的原代脂肪肝细胞,用靶向小鼠UCP-2基因的RNA干扰慢病毒载体感染所获得的脂肪肝细胞,实现对UCP-2基}q的特异性敲减(实验组),另以空载体感染的脂肪肝细胞为对照.荧光显微镜镜检确定感染效率,实时聚合酶链反应鉴定敲减效果.以肿瘤坏死因子-α(TNF-α)作用于感染后的肝细胞,24 h后以碘化丙啶染色,流式细胞仪检测细胞凋亡情况;Western印迹法检测凋亡蛋白酶-3(Caspase-3)

  16. Immune escape mutants of Highly Pathogenic Avian Influenza H5N1 selected using polyclonal sera: identification of key amino acids in the HA protein.

    Directory of Open Access Journals (Sweden)

    Ioannis Sitaras

    Full Text Available Evolution of Avian Influenza (AI viruses--especially of the Highly Pathogenic Avian Influenza (HPAI H5N1 subtype--is a major issue for the poultry industry. HPAI H5N1 epidemics are associated with huge economic losses and are sometimes connected to human morbidity and mortality. Vaccination (either as a preventive measure or as a means to control outbreaks is an approach that splits the scientific community, due to the risk of it being a potential driving force in HPAI evolution through the selection of mutants able to escape vaccination-induced immunity. It is therefore essential to study how mutations are selected due to immune pressure. To this effect, we performed an in vitro selection of mutants from HPAI A/turkey/Turkey/1/05 (H5N1, using immune pressure from homologous polyclonal sera. After 42 rounds of selection, we identified 5 amino acid substitutions in the Haemagglutinin (HA protein, most of which were located in areas of antigenic importance and suspected to be prone to selection pressure. We report that most of the mutations took place early in the selection process. Finally, our antigenic cartography studies showed that the antigenic distance between the selected isolates and their parent strain increased with passage number.

  17. Fusion protein predicted amino acid sequence of the first US avian pneumovirus isolate and lack of heterogeneity among other US isolates.

    Science.gov (United States)

    Seal, B S; Sellers, H S; Meinersmann, R J

    2000-02-01

    Avian pneumovirus (APV) was first isolated from turkeys in the west-central US following emergence of turkey rhinotracheitis (TRT) during 1996. Subsequently, several APV isolates were obtained from the north-central US. Matrix (M) and fusion (F) protein genes of these isolates were examined for sequence heterogeneity and compared with European APV subtypes A and B. Among US isolates the M gene shared greater than 98% nucleotide sequence identity with only one nonsynonymous change occurring in a single US isolate. Although the F gene among US APV isolates shared 98% nucleotide sequence identity, nine conserved substitutions were detected in the predicted amino acid sequence. The predicted amino acid sequence of the US APV isolate's F protein had 72% sequence identity to the F protein of APV subtype A and 71% sequence identity to the F protein of APV subtype B. This compares with 83% sequence identity between the APV subtype A and B predicted amino acid sequences of the F protein. The US isolates were phylogenetically distinguishable from their European counterparts based on F gene nucleotide or predicted amino acid sequences. Lack of sequence heterogeneity among US APV subtypes indicates these viruses have maintained a relatively stable population since the first outbreak of TRT. Phylogenetic analysis of the F protein among APV isolates supports classification of US isolates as a new APV subtype C.

  18. Avian influenza

    Directory of Open Access Journals (Sweden)

    Tjandra Y. Aditama

    2006-06-01

    Full Text Available Avian influenza, or “bird flu”, is a contagious disease of animals which crossed the species barrier to infect humans and gave a quite impact on public health in the world since 2004, especially due to the threat of pandemic situation. Until 1st March 2006, laboratory-confirmed human cases have been reported in seven countries: Cambodia, Indonesia, Thailand, Viet Nam, China, Iraq and Turkey with a total of 174 cases and 94 dead (54.02%. Indonesia has 27 cases, 20 were dead (74.07%. AI cases in Indonesia are more in male (62.5% and all have a symptom of fever. An influenza pandemic is a rare but recurrent event. An influenza pandemic happens when a new subtype emerges that has not previously circulated in humans. For this reason, avian H5N1 is a strain with pandemic potential, since it might ultimately adapt into a strain that is contagious among humans. Impact of the pandemic could include high rates of illness and worker absenteeism are expected, and these will contribute to social and economic disruption. Historically, the number of deaths during a pandemic has varied greatly. Death rates are largely determined by four factors: the number of people who become infected, the virulence of the virus, the underlying characteristics and vulnerability of affected populations, and the effectiveness of preventive measures. Accurate predictions of mortality cannot be made before the pandemic virus emerges and begins to spread. (Med J Indones 2006; 15:125-8Keywords: Avian Influenza, Pandemic

  19. Molecular patterns of avian influenza A viruses

    Institute of Scientific and Technical Information of China (English)

    KOU Zheng; LEI FuMin; WANG ShengYue; ZHOU YanHong; LI TianXian

    2008-01-01

    Avian influenza A viruses could get across the species barrier and be fatal to humans. Highly patho-genic avian influenza H5N1 virus was an example. The mechanism of interspecies transmission is not clear as yet. In this research, the protein sequences of 237 influenza A viruses with different subtypes were transformed into pseudo-signals. The energy features were extracted by the method of wavelet packet decomposition and used for virus classification by the method of hierarchical clustering. The clustering results showed that five patterns existed in avian influenza A viruses, which associated with the phenotype of interspecies transmission, and that avian viruses with patterns C and E could across species barrier and those with patterns A, B and D might not have the abilities. The results could be used to construct an early warning system to predict the transmissibility of avian influenza A viruses to humans.

  20. The amino-terminal region of the neuraminidase protein from avian H5N1 influenza virus is important for its biosynthetic transport to the host cell surface.

    Science.gov (United States)

    Qian, Guomin; Wang, Song; Chi, Xiaojuan; Li, Hua; Wei, Haitao; Zhu, Xiaomei; Chen, Yuhai; Chen, Ji-Long

    2014-12-01

    Influenza virus neuraminidase (NA) is a major viral envelope glycoprotein, which plays a critical role in viral infection. Although NA functional domains have been determined previously, the precise role of the amino acids located at the N-terminus of avian H5N1 NA for protein expression and intracellular transport to the host plasma membrane is not fully understood. In the present study, a series of N-terminal truncation or deletion mutants of H5N1 NA were generated and their expression and intracellular trafficking were investigated. Protein expression from mutants NAΔ20, NAΔ35, NAΔ40, NAΔ7-20 and NAΔ7-35 was undetectable by immunoblotting and by performing NA activity assays. Mutants NAΔ6, NAΔ11 and NAΔ15-20 showed a marked decreased in protein expression, whereas mutants NAΔ7-15 and NAΔ15 displayed a slight increase in protein expression, compared with that of the native NA protein. These data suggest that amino acid residues 16-20 are vital for NA protein expression, while amino acids 7-15 might suppress NA protein expression. In deletion mutants NAΔ7-15 and NAΔ15 there was an accumulation of NA protein at the juxta-nuclear region, with reduced expression of NA at the cell surface. Although active Cdc42 could promote transport of wild-type NA to the host cell surface, this member of the Rho family of GTPases failed to regulate transport of mutants NAΔ7-15 and NAΔ15. The results of the study reveal that amino acid residues 7-15 of H5N1 NA are critical for its biosynthetic transport to the host cell surface.

  1. D701N mutation in the PB2 protein contributes to the pathogenicity of H5N1 avian influenza viruses but not transmissibility in guinea pigs

    Directory of Open Access Journals (Sweden)

    Peirong eJiao

    2014-11-01

    Full Text Available H5N1 highly pathogenic avian influenza virus (HPAIV of clade 2.3.2 has been circulating in waterfowl in Southern China since 2003. Our previous studies showed that certain H5N1 HPAIV isolates within clade 2.3.2 from Southern China had high pathogenicity in different birds. Guinea pigs have been successfully used as models to evaluate the transmissibility of AIVs and other species of influenza viruses in mammalian hosts. However, few studies have reported pathogenicity and transmissibility of H5N1 HPAIVs of this clade in guinea pigs. In this study, we selected an H5N1 HPAIV isolate, A/duck/Guangdong/357/2008, to investigate the pathogenicity and transmissibility of the virus in guinea pigs. The virus had high pathogenicity in mice; additionally, it only replicated in some tissues of the guinea pigs without production of clinical signs, but was transmissible among guinea pigs. Interestingly, virus isolates from co-caged guinea pigs had the D701N mutation in the PB2 protein. These mutant viruses showed higher pathogenicity in mice and higher replication capability in guinea pigs but did not demonstrate enhanced the transmissibility among guinea pigs. These findings indicate the transmission of the H5N1 virus between mammals could induce virus mutations, and the mutant viruses might have higher pathogenicity in mammals without higher transmissibility. Therefore, the continued evaluation of the pathogenicity and transmissibility of avian influenza virus (AIVs in mammals is critical to the understanding of the evolutionary characteristics of AIVs and the emergence of potential pandemic strains.

  2. D701N mutation in the PB2 protein contributes to the pathogenicity of H5N1 avian influenza viruses but not transmissibility in guinea pigs.

    Science.gov (United States)

    Jiao, Peirong; Wei, Liangmeng; Song, Yafen; Cui, Jin; Song, Hui; Cao, Lan; Yuan, Runyu; Luo, Kaijian; Liao, Ming

    2014-01-01

    H5N1 highly pathogenic avian influenza virus (HPAIV) of clade 2.3.2 has been circulating in waterfowl in Southern China since 2003. Our previous studies showed that certain H5N1 HPAIV isolates within clade 2.3.2 from Southern China had high pathogenicity in different birds. Guinea pigs have been successfully used as models to evaluate the transmissibility of AIVs and other species of influenza viruses in mammalian hosts. However, few studies have reported pathogenicity and transmissibility of H5N1 HPAIVs of this clade in guinea pigs. In this study, we selected an H5N1 HPAIV isolate, A/duck/Guangdong/357/2008, to investigate the pathogenicity and transmissibility of the virus in guinea pigs. The virus had high pathogenicity in mice; additionally, it only replicated in some tissues of the guinea pigs without production of clinical signs, but was transmissible among guinea pigs. Interestingly, virus isolates from co-caged guinea pigs had the D701N mutation in the PB2 protein. These mutant viruses showed higher pathogenicity in mice and higher replication capability in guinea pigs but did not demonstrate enhanced the transmissibility among guinea pigs. These findings indicate the transmission of the H5N1 virus between mammals could induce virus mutations, and the mutant viruses might have higher pathogenicity in mammals without higher transmissibility. Therefore, the continued evaluation of the pathogenicity and transmissibility of avian influenza virus (AIVs) in mammals is critical to the understanding of the evolutionary characteristics of AIVs and the emergence of potential pandemic strains.

  3. Discovery of a novel functional leptin protein (LEP) in zebra finches: evidence for the existence of an authentic avian leptin gene predominantly expressed in the brain and pituitary.

    Science.gov (United States)

    Huang, Guian; Li, Juan; Wang, Hongning; Lan, Xinyu; Wang, Yajun

    2014-09-01

    Leptin (LEP) is reported to play important roles in controlling energy balance in vertebrates, including birds. However, it remains an open question whether an authentic "LEP gene" exists and functions in birds. Here, we identified and characterized a LEP gene (zebra finch LEP [zbLEP]) encoding a 172-amino acid precursor in zebra finches. Despite zbLEP showing limited amino acid sequence identity (26%-29%) to human and mouse LEPs, synteny analysis proved that zbLEP is orthologous to mammalian LEP. Using a pAH32 luciferase reporter system and Western blot analysis, we demonstrated that the recombinant zbLEP protein could potently activate finch and chicken LEP receptors (zbLEPR; cLEPR) expressed in human embryonic kidney 293 cells and enhance signal transducer and activator of transcription 3 phosphorylation, further indicating that zbLEP is a functional ligand for avian LEPRs. Interestingly, quantitative real-time RT-PCR revealed that zbLEP mRNA is expressed nearly exclusively in the pituitary and various brain regions but undetectable in adipose tissue and liver, whereas zbLEPR mRNA is widely expressed in adult finch tissues examined with abundant expression noted in pituitary, implying that unlike mammalian LEP, finch LEP may not act as an adipocyte-derived signal to control energy balance. As in finches, a LEP highly homologous to zbLEP was also identified in budgerigar genome. Strikingly, finch and budgerigar LEPs show little homology with chicken LEP (cLEP) previously reported, suggesting that the so-called cLEP is incorrect. Collectively, our data provide convincing evidence for the existence of an authentic functional LEP in avian species and suggest an important role of brain- and pituitary-derived LEP played in vertebrates.

  4. PTH stimulated growth and decreased Col-X deposition are phosphotidylinositol-3,4,5 triphosphate kinase and mitogen activating protein kinase dependent in avian sterna.

    Science.gov (United States)

    Harrington, Erik Kern; Coon, David J; Kern, Matthew F; Svoboda, Kathy K H

    2010-02-01

    Type X collagen (Col-X) deposition is a marker of terminal differentiation during chondrogenesis, in addition to appositional growth and apoptosis. The parathyroid hormone/parathyroid hormone related peptide (PTH/PTHrP) receptor, or PPR, is a G-Protein coupled receptor (GPCR), which activates several downstream pathways, moderating chondrocyte differentiation, including suppression of Col-X deposition. An Avian sterna model was used to analyze the PPR GPCR downstream kinase role in growth rate and extracellular matrix (ECM) including Col-II, IX, and X. Phosphatidylinositol kinase (PI3K), mitogen activating protein kinase (MAPK) and protein kinase A (PKA) were inhibited with specific established inhibitors LY294002, PD98059, and H89, respectively to test the hypothesis that they could reverse/inhibit the PTH/PTHrP pathway. Excised E14 chick sterna were PTH treated with or without an inhibitor and compared to controls. Sternal length was measured every 24 hr. Cultured sterna were immuno-stained using specific antibodies for Col-II, IX, or X and examined via confocal microscopy. Increased growth in PTH-treated sterna was MAPK, PI3K, and PKA dose dependent, suggesting growth was regulated through multiple pathways. Col-X deposition was rescued in PTH-treated sterna in the presence of PI3K or MAPK inhibitors, but not with the PKA inhibitor. All three inhibitors moderately disrupted Col-II and Col-IX deposition. These results suggest that PTH can activate multiple pathways during chondrocyte differentiation.

  5. Relationship between type 2 diabetes mellitus complicated by periodontal infection and uncoupling protein 3 gene%2型糖尿病并发牙周感染与解偶联蛋白3基因的关系研究

    Institute of Scientific and Technical Information of China (English)

    周逸丹; 张萍萍; 黄爱华

    2011-01-01

    目的 探讨2型糖尿病并发牙周感染与解偶联蛋白3(UCP3)基因的关系.方法 将80例2型糖尿病患者根据是否并发牙周感染分为两组,用聚合酶链反应(pCR)法对两组UCP3基因多态性进行分析并比较.结果 并发感染组II、ID、DD的基因频率分别为39.0%、51.2%、9.8%,与非并发感染组比较,差异无统计学意义;等位基因频率两组间亦差异无统计学意义.结论 2型糖尿病并发牙周感染与解偶联蛋白3基因之间未见明显的关系.%OBJECTIVE To explore the relationship between type 2 diabetes complicated by periodontal infection and uncoupling protein 3 (UCP3) gene.METHODS 80 patients with type 2 diabetes were divided into two groups according to whether complicated by periodontal infection.PCR method was used to analyze the polymorphism of UCP3 gene and the results were compared.RESULTS The gene frequencie of Ⅱ, ID, DD of the infection group was 39.0 %, 51.2 %, 9.8 %, respectively.There was no significant difference between the two groups(P>0.05).The allele frequency between the two groups had no significant difference (P>0.05).CONCLUSION It shows no significant relationship between type 2 diabetes complicated by periodontal infection and uncoupling protein 3 genes.

  6. 持续冷热环境对肉鸡生产性能、糖代谢和解偶联蛋白 mRNA表达的影响%Effects of Prolonged Cold and Hot Environment on Performance, Glucose Metabolism and Uncoupling Protein mRNA Expression of Broilers

    Institute of Scientific and Technical Information of China (English)

    苏红光; 张敏红; 冯京海; 吴鑫; 胡春红

    2014-01-01

    本试验研究了不同环境温度(10~30℃)持续14 d对肉鸡生产性能、糖代谢和禽类解偶联蛋白( avUCP) mRNA表达的影响。试验选取21日龄爱拔益加( AA)肉鸡288只,随机分到6个人工环境控制舱中,每个舱饲养6笼,每笼8只鸡作为1个重复。预试期7 d,温度22℃,相对湿度60%。28日龄时将各环境控制舱温度分别逐渐(1 h内)调到10、14、18、22、26和30℃,相对湿度60%,温湿度均保持恒定直至试验结束。正试期14 d。结果表明:1)试验期内,30℃组的体重( BW)显著低于14~26℃组( P<0.05);22~30℃组平均日采食量( ADFI)随温度升高而显著下降( P<0.05);10~30℃组平均日增重( ADG)随温度升高出现先升高后下降的趋势,在22℃时最高;料重比( F/G)随温度升高呈现先下降后升高的趋势,22℃时最低;平均日饮水量( ADWC)在10℃组最低。2)试验第14天,26℃组血糖水平显著低于18℃组( P<0.05);肝糖原水平在各组之间差异不显著( P>0.05);22℃组肌糖原水平显著低于10、26和30℃组( P<0.05)。3)试验第14天,18、22℃组avUCP mRNA相对表达量显著高于其他组( P<0.05)。结果提示:在本试验条件下,从生产性能和能量利用效率考虑,28~42日龄AA肉鸡的适宜养殖温度为22~26℃。%This study was conducted to investigate the effects of different environmental temperatures ( ranged from 10 to 30 ℃) on performance, glucose metabolism and expression of avian uncoupling protein ( avUCP) mRNA of broilers. Two hundred and eighty eight 21-day-old Arbor Acres ( AA) broilers were assigned to six environmental chambers, each chamber contained six cages with eight birds per cage, and each cage as a repli-cate. The pre-test period lasted for 7 days and broilers were kept at 22℃ temperature and 60% relative humidi-ty. When broilers were

  7. The PSAP motif within the ORF3 protein of an avian strain of the hepatitis E virus is not critical for viral infectivity in vivo but plays a role in virus release.

    Science.gov (United States)

    Kenney, Scott P; Pudupakam, R S; Huang, Yao-Wei; Pierson, F William; LeRoith, Tanya; Meng, Xiang-Jin

    2012-05-01

    The ORF3 protein of hepatitis E virus (HEV) is a multifunctional protein important for virus replication. The ORF3 proteins from human, swine, and avian strains of HEV contain a conserved PXXP amino acid motif, resembling either Src homology 3 (SH3) cell signaling interaction motifs or "late domains" involved in host cell interactions aiding in particle release. Using an avian strain of HEV, we determined the roles of the conserved prolines within the PREPSAPP motif in HEV replication and infectivity in Leghorn male hepatoma (LMH) chicken liver cells and in chickens. Each proline was changed to alanine to produce 8 avian HEV mutants containing single mutations (P64, P67, P70, and P71 to A), double mutations (P64/67A, P64/70A, and P67/70A), and triple mutations (P64/67/70A). The results showed that avian HEV mutants are replication competent in vitro, and none of the prolines in the PXXPXXPP motif are essential for infectivity in vivo; however, the second and third prolines appear to aid in fecal virus shedding, suggesting that the PSAP motif, but not the PREP motif, is involved in virus release. We also showed that the PSAP motif interacts with the host protein tumor suppressor gene 101 (TSG101) and that altering any proline within the PSAP motif disrupts this interaction. However, we showed that the ORF2 protein expressed in LMH cells is efficiently released from the cells in the absence of ORF3 and that coexpression of ORF2 and ORF3 did not act synergistically in this release, suggesting that another factor(s) such as ORF1 or viral genomic RNA may be necessary for proper particle release.

  8. Non-avian animal reservoirs present a source of influenza A PB1-F2 proteins with novel virulence-enhancing markers.

    Directory of Open Access Journals (Sweden)

    Irina V Alymova

    Full Text Available PB1-F2 protein, expressed from an alternative reading frame of most influenza A virus (IAV PB1 segments, may possess specific residues associated with enhanced inflammation (L62, R75, R79, and L82 and cytotoxicity (I68, L69, and V70. These residues were shown to increase the pathogenicity of primary viral and secondary bacterial infections in a mouse model. In contrast to human seasonal influenza strains, virulence-associated residues are present in PB1-F2 proteins from pandemic H1N1 1918, H2N2 1957, and H3N2 1968, and highly pathogenic H5N1 strains, suggesting their contribution to viruses' pathogenic phenotypes. Non-human influenza strains may act as donors of virulent PB1-F2 proteins. Previously, avian influenza strains were identified as a potential source of inflammatory, but not cytotoxic, PB1-F2 residues. Here, we analyze the frequency of virulence-associated residues in PB1-F2 sequences from IAVs circulating in mammalian species in close contact with humans: pigs, horses, and dogs. All four inflammatory residues were found in PB1-F2 proteins from these viruses. Among cytotoxic residues, I68 was the most common and was especially prevalent in equine and canine IAVs. Historically, PB1-F2 from equine (about 75% and canine (about 20% IAVs were most likely to have combinations of the highest numbers of residues associated with inflammation and cytotoxicity, compared to about 7% of swine IAVs. Our analyses show that, in addition to birds, pigs, horses, and dogs are potentially important sources of pathogenic PB1-F2 variants. There is a need for surveillance of IAVs with genetic markers of virulence that may be emerging from these reservoirs in order to improve pandemic preparedness and response.

  9. Nucleotide sequences of the F, L and G protein genes of two non-A/non-B avian pneumoviruses (APV) reveal a novel APV subgroup.

    Science.gov (United States)

    Bäyon-Auboyer, M H; Arnauld, C; Toquin, D; Eterradossi, N

    2000-11-01

    Sequence analysis was performed of all or part of the genes encoding the fusion (F), polymerase (L) and attachment (G) proteins of two French non-A/non-B avian pneumovirus (APV) isolates (Fr/85/1 and Fr/85/2). The two isolates shared at least 99.7% nt and 99.0% aa sequence identity. Comparison with the F genes from subgroup A, subgroup B or Colorado APVs revealed nt and aa identities of 70.0-80. 5% and 77.6-97.2%, respectively, with the L gene sharing 76.1% nt and 85.3% aa identity with that of a subgroup A isolate. The Fr/85/1 and Fr/85/2 G genes comprised 1185 nt, encoding a protein of 389 aa. Common features with subgroup A and subgroup B G proteins included an amino-terminal membrane anchor, a high serine and threonine content, conservation of cysteine residues and a single extracellular region of highly conserved sequence proposed to be the functional domain involved in virus attachment to cellular receptors. However, the Fr/85/1 and Fr/85/2 G sequences shared at best 56.6% nt and 31.2% aa identity with subgroup A and B APVs, whereas these isolates share 38% aa identity. Phylogenetic analysis of the F, G and L genes of pneumoviruses suggested that isolates Fr/85/1 and Fr/85/2 belong to a previously unrecognized APV subgroup, tentatively named D. G-based oligonucleotide primers were defined for the specific molecular identification of subgroup D. These are the first G protein sequences of non-A/non-B APVs to be determined.

  10. Proteomic Analysis of Human Brown Adipose Tissue Reveals Utilization of Coupled and Uncoupled Energy Expenditure Pathways.

    Science.gov (United States)

    Müller, Sebastian; Balaz, Miroslav; Stefanicka, Patrik; Varga, Lukas; Amri, Ez-Zoubir; Ukropec, Jozef; Wollscheid, Bernd; Wolfrum, Christian

    2016-07-15

    Human brown adipose tissue (BAT) has become an attractive target to combat the current epidemical spread of obesity and its associated co-morbidities. Currently, information on its functional role is primarily derived from rodent studies. Here, we present the first comparative proteotype analysis of primary human brown adipose tissue versus adjacent white adipose tissue, which reveals significant quantitative differences in protein abundances and in turn differential functional capabilities. The majority of the 318 proteins with increased abundance in BAT are associated with mitochondrial metabolism and confirm the increased oxidative capacity. In addition to uncoupling protein 1 (UCP1), the main functional effector for uncoupled respiration, we also detected the mitochondrial creatine kinases (CKMT1A/B, CKMT2), as effective modulators of ATP synthase coupled respiration, to be exclusively expressed in BAT. The abundant expression and utilization of both energy expenditure pathways in parallel highlights the complex functional involvement of BAT in human physiology.

  11. Effects of acute and chronic endurance exercise on mitochondrial uncoupling in human skeletal muscle

    DEFF Research Database (Denmark)

    Fernström, Maria; Tonkonogi, Michail; Sahlin, Kent

    2004-01-01

    Mitochondrial proteins such as uncoupling protein 3 (UCP3) and adenine nucleotide translocase (ANT) may mediate back-leakage of protons and serve as uncouplers of oxidative phosphorylation. We hypothesized that UCP3 and ANT increase after prolonged exercise and/or endurance training, resulting...... respiration or state 3). Protein expression of UCP3 and ANT was measured with Western blotting. After endurance training, .VO2peak, citrate synthase activity (CS), state 3 respiration and ANT increased by 24, 47, 40 and 95%, respectively (all P UCP3 remained unchanged. When expressed per unit...... of CS (a marker of mitochondrial volume) UCP3 and UCR decreased by 54% and 18%(P 3 h of recovery (P

  12. Cascaded uncoupled dual-ring modulator

    CERN Document Server

    Gu, Tingyi; Wong, Chee Wei; Dong, Po

    2014-01-01

    We demonstrate that by coherent driving two uncoupled rings in same direction, the effective photon circulating time in the dual ring modulator is reduced, with increased modulation quality. The inter-ring detuning dependent photon dynamics, Q-factor, extinction ratio and optical modulation amplitude of two cascaded silicon ring resonators are studied and compared with that of a single ring modulator. Experimentally measured eye diagrams, together with coupled mode theory simulations, demonstrate the enhancement of dual ring configuration at 20 Gbps with a Q ~ 20,000.

  13. Optimal parameters uncoupling vibration modes of oscillators

    CERN Document Server

    Le, Khanh Chau

    2016-01-01

    A novel optimization concept for an oscillator with two degrees of freedom is proposed. By using specially defined motion ratios, we control the action of springs and dampers to each degree of freedom of the oscillator. If the potential action of the springs in one period of vibration, used as the payoff function for the conservative oscillator, is maximized, then the optimal motion ratios uncouple vibration modes. The same result holds true for the dissipative oscillator. The application to optimal design of vehicle suspension is discussed.

  14. Uncoupling of oxidative phosphorylation by curcumin: Implication of its cellular mechanism of action

    Energy Technology Data Exchange (ETDEWEB)

    Lim, Han Wern; Lim, Hwee Ying [Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119260 (Singapore); Wong, Kim Ping, E-mail: bchsitkp@nus.edu.sg [Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119260 (Singapore)

    2009-11-06

    Curcumin is a phytochemical isolated from the rhizome of turmeric. Recent reports have shown curcumin to have antioxidant, anti-inflammatory and anti-tumor properties as well as affecting the 5'-AMP activated protein kinase (AMPK), mTOR and STAT-3 signaling pathways. We provide evidence that curcumin acts as an uncoupler. Well-established biochemical techniques were performed on isolated rat liver mitochondria in measuring oxygen consumption, F{sub 0}F{sub 1}-ATPase activity and ATP biosynthesis. Curcumin displays all the characteristics typical of classical uncouplers like fccP and 2,4-dinitrophenol. In addition, at concentrations higher than 50 {mu}M, curcumin was found to inhibit mitochondrial respiration which is a characteristic feature of inhibitory uncouplers. As a protonophoric uncoupler and as an activator of F{sub 0}F{sub 1}-ATPase, curcumin causes a decrease in ATP biosynthesis in rat liver mitochondria. The resulting change in ATP:AMP could disrupt the phosphorylation status of the cell; this provides a possible mechanism for its activation of AMPK and its downstream mTOR and STAT-3 signaling.

  15. Rol de las proteinas desacoplantes UCP1, UCP2 y UCP3 en el gasto energetico, diabetes tipo 2 y obesidad: Sinergismo con la tiroides Role of uncoupling proteins UCP1, UCP2 and UCP3 in energy balance, type 2 diabetes and obesity: Synergism with the thyroid

    Directory of Open Access Journals (Sweden)

    Ángel A. Zaninovich

    2005-04-01

    and dissipation through the action of uncoupling protein-1 (UCP1 during cold stress, showed the relevance of this tissue for energy expenditure in lower mammals. UCP1 is only expressed in BAT through the synergistic action of norepinephrine (NE and thyroid hormones in animals exposed to cold and to a lesser degree after meals. The uncoupling protein-2 (UCP2 is found in many tissues and exerts dual effects: it protects cells function from damage caused by reactive oxygen species (ROS. On the other hand, the uncoupling induced by UCP2 in mitochondria of pancreatic ß cells decreases ATP síntesis and impairs insulin secretion in response to glucose. Hyperlipidemia also prevents insulin secretion through a similar pathway, leading to hyperglycemia. The uncoupling protein-3 is found mostly in skeletal muscle and BAT and its absence did not alter heat production or body temperature. This protein would export fatty acids ouside the mitochondrial matrix for combustion in tissues where fat is the main fuel. In humans, the uncoupling proteins may not play a leading role in energy regulation. However, intensive studies on these and other factors influencing energy expenditure, appetite and glucose metabolism are taken place worldwide and may soon provide more clues on the mechanisms regulating energy balance and their use in the prevention or treatment of human obesity and diabetes type 2.

  16. INDUCTION OF ANTIVIRAL IMMUNE-RESPONSES BY IMMUNIZATION WITH RECOMBINANT-DNA ENCODED AVIAN CORONAVIRUS NUCLEOCAPSID PROTEIN

    NARCIS (Netherlands)

    BOOTS, AMH; BENAISSATROUW, BJ; HESSELINK, W; RIJKE, E; SCHRIER, C; HENSEN, EJ; Boots, Annemieke

    1992-01-01

    Immune responses to the infectious bronchitis virus (IBV) nucleocapsid protein were studied using a recombinant-DNA expression product. In mice, a lymphocyte proliferative response and a delayed-type hypersensitivity reaction to IBV were induced upon immunization with this nucleocapsid protein. Next

  17. [Detection and description of avian hepatitis E virus isolated in China--a review].

    Science.gov (United States)

    Zhao, Qin; Sun, Yani; Zhou, Enmin

    2012-03-04

    Avian hepatitis E virus (HEV), a member of Hepeviridae family, is genetically and antigenically related with human and swine HEV in the family. Since its discovery, avian HEV infection has been investigated in many countries from serology and molecular epidemiology studies. At present, five complete or near complete genomes of avian HEV isolates were reported in GenBank and were divided into three genotypes. The complete genome of avian HEV contains 3 ORFs of which ORF2 gene encodes capsid protein containing the primary epitopes of viral particles and is target gene for serodiagnostic antigen and vaccine candidate. Because avian HEV infection has significant impact on the poultry industry and potential zoonotic transmission, the researches on avian HEV have been given much attention. We here give a broad review of the research update on the aetiology, pathogenesis and the antigenicity of capsid protein of avian HEV based on identification of Chinese avian HEV isolate.

  18. Avian And Other Zoonotic Influenza

    Science.gov (United States)

    ... files Questions & answers Features Multimedia Contacts Avian and other zoonotic influenza Fact sheet Updated November 2016 Key ... A(H3) subtypes. Clinical features of avian and other zoonotic influenza infections in humans Avian and other ...

  19. Multimeric recombinant M2e protein-based ELISA: a significant improvement in differentiating avian influenza infected chickens from vaccinated ones.

    Directory of Open Access Journals (Sweden)

    Farshid Hadifar

    Full Text Available Killed avian influenza virus (AIV vaccines have been used to control H5N1 infections in countries where the virus is endemic. Distinguishing vaccinated from naturally infected birds (DIVA in such situations however, has become a major challenge. Recently, we introduced the recombinant ectodomain of the M2 protein (M2e of H5N1 subtype as a novel tool for an ELISA based DIVA test. Despite being antigenic in natural infection the monomer form of the M2e used in ELISA had limited antigenicity and consequently poor diagnostic capability. To address this shortcoming, we evaluated the use of four tandem copies of M2e (tM2e for increased efficiency of M2e antibody detection. The tM2e gene of H5N1 strain from Indonesia (A/Indonesia/CDC540/2006 was cloned into a pMAL- p4x expression vector and expressed in E.coli as a recombinant tM2e-MBP or M2e-MBP proteins. Both of these, M2e and tM2e antigens reacted with sera obtained from chickens following live H5N1 infection but not with sera from vaccinated birds. A significantly stronger M2e antibody reaction was observed with the tM2e compared to M2e antigen. Western blotting also supported the superiority of tM2e over M2e in detection of specific M2e antibodies against live H5N1 infection. Results from this study demonstrate that M2e tetramer is a better antigen than single M2e and could be more suitable for an ELISA based DIVA test.

  20. Avian amyloidosis

    NARCIS (Netherlands)

    Landman, W.J.M.; Gruys, E.; Gielkens, A.L.J.

    1998-01-01

    Although amyloid deposits have been described for more than a century and a half, its proteinaceous and fibrillar nature was not revealed until after 1950. Biochemical characterization of amyloids has brought to light that several non-related proteins can re-organize into amyloid fibrils. In some do

  1. Skeletal muscle mitochondrial uncoupling drives endocrine cross-talk through induction of FGF21 as a myokine

    NARCIS (Netherlands)

    Keipert, S.; Ost, M.; Johann, K.; Imber, F.; Jastroch, M.; Schothorst, van E.M.; Keijer, J.; Klaus, S.

    2014-01-01

    UCP1-Tg mice with ectopic expression of uncoupling protein 1 (UCP1) in skeletal muscle (SM) are a model of improved substrate metabolism and increased longevity. Analysis of myokine expression showed an induction of fibroblast growth factor 21 (FGF21) in SM, resulting in approximately fivefold eleva

  2. A novel hemagglutinin protein produced in bacteria protects chickens against H5N1 highly pathogenic avian influenza viruses by inducing H5 subtype-specific neutralizing antibodies

    Science.gov (United States)

    Sączyńska, Violetta; Romanik, Agnieszka; Florys, Katarzyna; Cecuda-Adamczewska, Violetta; Kęsik-Brodacka, Małgorzata; Śmietanka, Krzysztof; Olszewska, Monika; Domańska-Blicharz, Katarzyna; Minta, Zenon; Szewczyk, Bogusław; Płucienniczak, Grażyna; Płucienniczak, Andrzej

    2017-01-01

    The highly pathogenic (HP) H5N1 avian influenza viruses (AIVs) cause a mortality rate of up to 100% in infected chickens and pose a permanent pandemic threat. Attempts to obtain effective vaccines against H5N1 HPAIVs have focused on hemagglutinin (HA), an immunodominant viral antigen capable of eliciting neutralizing antibodies. The vast majority of vaccine projects have been performed using eukaryotic expression systems. In contrast, we used a bacterial expression system to produce vaccine HA protein (bacterial HA) according to our own design. The HA protein with the sequence of the H5N1 HPAIV strain was efficiently expressed in Escherichia coli, recovered in the form of inclusion bodies and refolded by dilution between two chromatographic purification steps. Antigenicity studies showed that the resulting antigen, referred to as rH5-E. coli, preserves conformational epitopes targeted by antibodies specific for H5-subtype HAs, inhibiting hemagglutination and/or neutralizing influenza viruses in vitro. The proper conformation of this protein and its ability to form functional oligomers were confirmed by a hemagglutination test. Consistent with the biochemical characteristics, prime-boost immunizations with adjuvanted rH5-E. coli protected 100% and 70% of specific pathogen-free, layer-type chickens against challenge with homologous and heterologous H5N1 HPAIVs, respectively. The observed protection was related to the positivity in the FluAC H5 test (IDVet) but not to hemagglutination-inhibiting antibody titers. Due to full protection, the effective contact transmission of the homologous challenge virus did not occur. Survivors from both challenges did not or only transiently shed the viruses, as established by viral RNA detection in oropharyngeal and cloacal swabs. Our results demonstrate that vaccination with rH5-E. coli could confer control of H5N1 HPAIV infection and transmission rates in chicken flocks, accompanied by reduced virus shedding. Moreover, the role of

  3. Uncoupling and oxidative stress in liver mitochondria isolated from rats with acute iron overload

    Energy Technology Data Exchange (ETDEWEB)

    Pardo Andreu, G.L. [Centro de Quimica Farmaceutica, Departamento de Investigaciones Biomedicas, Ciudad de La Habana (Cuba); Inada, N.M.; Vercesi, A.E. [Universidade Estadual de Campinas, Departamento de Patologia Clinica, Faculdade de Ciencias Medicas, Campinas, SP (Brazil); Curti, C. [Universidade de Sao Paulo, Departamento de Fisica e Quimica, Faculdade de Ciencias Farmaceuticas de Ribeirao Preto, SP (Brazil)

    2009-01-15

    One hypothesis for the etiology of cell damage arising from iron overload is that its excess selectively affects mitochondria. Here we tested the effects of acute iron overload on liver mitochondria isolated from rats subjected to a single dose of i.p. 500 mg/kg iron-dextran. The treatment increased the levels of iron in mitochondria (from 21{+-}4 to 130{+-}7 nmol/mg protein) and caused both lipid peroxidation and glutathione oxidation. The mitochondria of iron-treated rats showed lower respiratory control ratio in association with higher resting respiration. The mitochondrial uncoupling elicited by iron-treatment did not affect the phosphorylation efficiency or the ATP levels, suggesting that uncoupling is a mitochondrial protective mechanism against acute iron overload. Therefore, the reactive oxygen species (ROS)/H{sup +} leak couple, functioning as a mitochondrial redox homeostatic mechanism could play a protective role in the acutely iron-loaded mitochondria. (orig.)

  4. The ectodomains but not the transmembrane domains of the fusion proteins of subtypes A and B avian pneumovirus are conserved to a similar extent as those of human respiratory syncytial virus.

    Science.gov (United States)

    Naylor, C J; Britton, P; Cavanagh, D

    1998-06-01

    The fusion glycoprotein (F(B)) gene of five strains of the B subtype of avian pneumovirus (APV; turkey rhinotracheitis virus) has been sequenced. The length of the F(B) protein was 538 amino acids, identical to that of the F protein of subtype A virus, with which it had 74% and 83% overall nucleotide and deduced amino acid identities, respectively. The F(B) and F(A) ectodomains had 90% amino acid identity, very similar to the 91% identity between the ectodomains of the F proteins of subtype A and B human respiratory syncytial virus (HRSV). As with HRSV, the F2 polypeptide was less conserved (83% identity) than F1 (94%). In contrast to the ectodomain, the transmembrane and cytoplasmic domains of the two APV subtypes were much less conserved (30% and 48% identity, respectively) than those of HRSV (92% and 87%, respectively). Comparisons within all the genera of the Paramyxoviridae (Pneumovirus, Morbillivirus, Paramyxovirus and Rubullavirus) show that low amino acid identity between F protein transmembrane domains is a feature of different species of virus rather than of strain differences. This may indicate that the two subtypes of APV have evolved in different geographical regions and/or different avian species. This is the first report of an F gene sequence from a subtype B APV.

  5. Avian Chlamydiosis Zoonotic Disease.

    Science.gov (United States)

    Szymańska-Czerwińska, Monika; Niemczuk, Krzysztof

    2016-01-01

    This review presents recent data about avian chlamydiosis. Chlamydia psittaci has been considered to be the main causative agent of chlamydiosis in birds; however, two new Chlamydia species have been detected recently-C. gallinacea in breeding birds and C. avium in wild birds. We discuss the zoonotic potential of avian Chlamydia species.

  6. Envelope and pre-membrane protein structural amino acid mutations mediate diminished avian growth and virulence of a Mexican West Nile virus isolate.

    Science.gov (United States)

    Langevin, Stanley A; Bowen, Richard A; Ramey, Wanichaya N; Sanders, Todd A; Maharaj, Payal D; Fang, Ying; Cornelius, Jennine; Barker, Christopher M; Reisen, William K; Beasley, David W C; Barrett, Alan D T; Kinney, Richard M; Huang, Claire Y-H; Brault, Aaron C

    2011-12-01

    The hallmark attribute of North American West Nile virus (WNV) strains has been high pathogenicity in certain bird species. Surprisingly, this avian virulent WNV phenotype has not been observed during its geographical expansion into the Caribbean, Central America and South America. One WNV variant (TM171-03-pp1) isolated in Mexico has demonstrated an attenuated phenotype in two widely distributed North American bird species, American crows (AMCRs) and house sparrows (HOSPs). In order to identify genetic determinants associated with attenuated avian replication of the TM171-03-pp1 variant, chimeric viruses between the NY99 and Mexican strains were generated, and their replicative capacity was assessed in cell culture and in AMCR, HOSP and house finch avian hosts. The results demonstrated that mutations in both the pre-membrane (prM-I141T) and envelope (E-S156P) genes mediated the attenuation phenotype of the WNV TM171-03-pp1 variant in a chicken macrophage cell line and in all three avian species assayed. Inclusion of the prM-I141T and E-S156P TM171-03-pp1 mutations in the NY99 backbone was necessary to achieve the avian attenuation level of the Mexican virus. Furthermore, reciprocal incorporation of both prM-T141I and E-P156S substitutions into the Mexican virus genome was necessary to generate a virus that exhibited avian virulence equivalent to the NY99 virus. These structural changes may indicate the presence of new evolutionary pressures exerted on WNV populations circulating in Latin America or may signify a genetic bottleneck that has constrained their epiornitic potential in alternative geographical locations.

  7. Avian Influenza infection in Human

    Directory of Open Access Journals (Sweden)

    Mohan. M

    2008-08-01

    infection is being demonstrated right now in Asia. However, urgent control of all outbreaks of avian influenza in birds - even when caused by a strain of low pathogenicity- is of utmost importance. Research has shown that certain, avian influenza virus strains, usually of low pathogenicity can rapidly Avian Influenza infection in Human mutate (within 6 to 9 months into a highly pathogenic strain if allowed to circulate in poultry populations. Altogether, more than half of the laboratoryconfirmed cases have been fatal. H5N1 avian influenza in humans is still a rare disease, but a severe one that must be closely watched and studied, particularly because of the potential of this virus to evolve in ways that could start a pandemic. The challenge for all of us is to gain an under-standing of how just 10 or 11 proteins of these viruses to replicate and be transmitted not only between hosts of one species but also between species. [Veterinary World 2008; 1(4.000: 122-125

  8. Uncoupling RARA transcriptional activation and degradation clarifies the bases for APL response to therapies.

    Science.gov (United States)

    Ablain, Julien; Leiva, Magdalena; Peres, Laurent; Fonsart, Julien; Anthony, Elodie; de Thé, Hugues

    2013-04-01

    In PML/RARA-driven acute promyelocytic leukemia (APL), retinoic acid (RA) induces leukemia cell differentiation and transiently clears the disease. Molecularly, RA activates PML/RARA-dependent transcription and also initiates its proteasome-mediated degradation. In contrast, arsenic, the other potent anti-APL therapy, only induces PML/RARA degradation by specifically targeting its PML moiety. The respective contributions of RA-triggered transcriptional activation and proteolysis to clinical response remain disputed. Here, we identify synthetic retinoids that potently activate RARA- or PML/RARA-dependent transcription, but fail to down-regulate RARA or PML/RARA protein levels. Similar to RA, these uncoupled retinoids elicit terminal differentiation, but unexpectedly fail to impair leukemia-initiating activity of PML/RARA-transformed cells ex vivo or in vivo. Accordingly, the survival benefit conferred by uncoupled retinoids in APL mice is dramatically lower than the one provided by RA. Differentiated APL blasts sorted from uncoupled retinoid-treated mice retain PML/RARA expression and reinitiate APL in secondary transplants. Thus, differentiation is insufficient for APL eradication, whereas PML/RARA loss is essential. These observations unify the modes of action of RA and arsenic and shed light on the potency of their combination in mice or patients.

  9. Muscle mitochondrial uncoupling dismantles neuromuscular junction and triggers distal degeneration of motor neurons.

    Directory of Open Access Journals (Sweden)

    Luc Dupuis

    Full Text Available BACKGROUND: Amyotrophic lateral sclerosis (ALS, the most frequent adult onset motor neuron disease, is associated with hypermetabolism linked to defects in muscle mitochondrial energy metabolism such as ATP depletion and increased oxygen consumption. It remains unknown whether muscle abnormalities in energy metabolism are causally involved in the destruction of neuromuscular junction (NMJ and subsequent motor neuron degeneration during ALS. METHODOLOGY/PRINCIPAL FINDINGS: We studied transgenic mice with muscular overexpression of uncoupling protein 1 (UCP1, a potent mitochondrial uncoupler, as a model of muscle restricted hypermetabolism. These animals displayed age-dependent deterioration of the NMJ that correlated with progressive signs of denervation and a mild late-onset motor neuron pathology. NMJ regeneration and functional recovery were profoundly delayed following injury of the sciatic nerve and muscle mitochondrial uncoupling exacerbated the pathology of an ALS animal model. CONCLUSIONS/SIGNIFICANCE: These findings provide the proof of principle that a muscle restricted mitochondrial defect is sufficient to generate motor neuron degeneration and suggest that therapeutic strategies targeted at muscle metabolism might prove useful for motor neuron diseases.

  10. Infection of Avian Pox Virus in Oriental Turtle-Doves

    Directory of Open Access Journals (Sweden)

    Kyung-Yeon Eo1, Young-Hoan Kim2, Kwang-Hyun Cho3, Jong-Sik Jang4, Tae-Hwan Kim5, Dongmi Kwak5 and Oh-Deog Kwon5*

    2011-10-01

    Full Text Available Three Oriental Turtle-doves (Streptopelia orientalis exhibiting lethargy, dyspnea, poor physical condition, and poor flight endurance, were rescued and referred to the Animal Health Center, Seoul Zoo, Korea. The doves had wart-like lesions on the legs and head. All of them died the following day after arrival, with the exception of one that survived for 6 days. Diphtheritic membranes on the tongue and oral mucosa were apparent at necropsy. Avian pox virus infection was suspected based on the proliferative skin lesions and oral diphtheritic lesions. Infection of the avian pox virus was confirmed by PCR using primers specific to the 4b core protein gene of avian pox virus. All cases were diagnosed with avian pox virus infection. This is believed to be the first description on natural infection of avian pox in Oriental Turtle-doves in Korea.

  11. Uncoupled thermoelasticity solutions applied on beam dumps

    Science.gov (United States)

    Ouzia, A.; Antonakakis, T.

    2016-06-01

    In particle accelerators the process of beam absorption is vital. At CERN particle beams are accelerated at energies of the order of TeV. In the event of a system failure or following collisions, the beam needs to be safely absorbed by dedicated protecting blocks. The thermal shock caused by the rapid energy deposition within the absorbing block causes thermal stresses that may rise above critical levels. The present paper provides a convenient expression of such stresses under hypotheses described hereafter. The temperature field caused by the beam energy deposition is assumed to be Gaussian. Such a field models a non-diffusive heat deposition. These effects are described as thermoelastic as long as the stresses remain below the proportional limit and can be analytically modeled by the coupled equations of thermoelasticity. The analytical solution to the uncoupled thermoelastic problem in an infinite domain is presented herein and matched with a finite unit radius sphere. The assumption of zero diffusion as well as the validity of the match with a finite geometry is quantified such that the obtained solutions can be rigorously applied to real problems. Furthermore, truncated series solutions, which are not novel, are used for comparison purposes. All quantities are nondimensional and the problem reduces to a dependence of five dimensionless parameters. The equations of elasticity are presented in the potential formulation where the shear potential is assumed to be nil due to the source being a gradient and the absence of boundaries. Nevertheless equivalent three-dimensional stresses are computed using the compressive potential and optimized using standard analytical optimization methods. An alternative algorithm for finding the critical points of the three-dimensional stress function is presented. Finally, a case study concerning the proton synchrotron booster dump is presented where the aforementioned analytical solutions are used and the preceding assumptions

  12. Editorial: Avian Research

    Institute of Scientific and Technical Information of China (English)

    Yong; Wang; Guangmei; Zheng

    2014-01-01

    <正>Welcome to Avian Research!This new journal is a continuation and enhancement of Chinese Birds,which has been and continues to be sponsored by the China Ornithological Society and Beijing Forestry University.In the four years since its inception,the original journal—the only one in China focusing on avian research—has published over 130 manuscripts,with authors from all continents across the world,garnering global respect in

  13. A Study on the Variance of Uncoupling Protein 3 Gene in Shanghai Han Race%上海地区汉族人解偶联蛋白3基因的变异

    Institute of Scientific and Technical Information of China (English)

    张蓉; 郑以漫; 项坤三; 贾伟平; 方启晨

    2001-01-01

    Objective To investigatce the relationship between ⅣS6 g→a + 1 variance of un- coupling protein 3 gene (UCP3) and obesity as well as diabetes in Chinese Hans. Methods The geno- type of this variance was determined by the PCR-RFLP method in 306 unrelated Shanghai Chinese Hans and 66 normal Caucasians. Results No ⅣS6 g→a + 1 variance of UCP3 gene was found in this group of Shanghai Chinese Hans and 66 Caucasians. Conclusion This variance of UCP3 gene is not a major predisposing genetic factor for obesity and diabetes in Chinese Hans.%目的探讨解偶联蛋白P3(UCP3)基因Ⅳ6G→a+1变异与中国汉族人肥胖及糖尿病的关系。方法选取306例无亲缘关系的中国人和66例白种人,运用PCR-RFLP方法检测UCP3基因Ⅳ两Pa+1的变异情况。结果中国人与白种人中均未发现这种变异。结论该基因变异不是中国汉族人肥胖与糖尿病发生的重要遗传因素。

  14. eNOS-uncoupling in age-related erectile dysfunction

    Science.gov (United States)

    Johnson, JM; Bivalacqua, TJ; Lagoda, GA; Burnett, AL; Musicki, B

    2011-01-01

    Aging is associated with ED. Although age-related ED is attributed largely to increased oxidative stress and endothelial dysfunction in the penis, the molecular mechanisms underlying this effect are not fully defined. We evaluated whether endothelial nitric oxide synthase (eNOS) uncoupling in the aged rat penis is a contributing mechanism. Correlatively, we evaluated the effect of replacement with eNOS cofactor tetrahydrobiopterin (BH4) on erectile function in the aged rats. Male Fischer 344 ‘young’ (4-month-old) and ‘aged’ (19-month-old) rats were treated with a BH4 precursor sepiapterin (10 mg/kg intraperitoneally) or vehicle for 4 days. After 1-day washout, erectile function was assessed in response to electrical stimulation of the cavernous nerve. Endothelial dysfunction (eNOS uncoupling) and oxidative stress (thiobarbituric acid reactive substances, TBARS) were measured by conducting western blot in penes samples. Erectile response was significantly reduced in aged rats, whereas eNOS uncoupling and TBARS production were significantly increased in the aged rat penis compared with young rats. Sepiapterin significantly improved erectile response in aged rats and prevented increase in TBARS production, but did not affect eNOS uncoupling in the penis of aged rats. These findings suggest that aging induces eNOS uncoupling in the penis, resulting in increased oxidative stress and ED. PMID:21289638

  15. Investigating the role of uncoupling of Troponin I phosphorylation from changes in myofibrillar Ca2+-sensitivity in the pathogenesis of Cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Andrew Easton Messer

    2014-08-01

    Full Text Available Contraction in the mammalian heart is controlled by the intracellular Ca2+ concentration as it is in all striated muscle, but the heart has an additional signalling system that comes into play to increase heart rate and cardiac output during exercise or stress. β-adrenergic stimulation of heart muscle cells leads to release of cyclic-AMP and the activation of protein kinase A which phosphorylates key proteins in the sarcolemma, sarcoplasmic reticulum and contractile apparatus. Troponin I (TnI and Myosin Binding Protein C (MyBP-C are the prime targets in the myofilaments. TnI phosphorylation lowers myofibrillar Ca2+-sensitivity and increases the speed of Ca2+-dissociation and relaxation (lusitropic effect.Recent studies have shown that this relationship between Ca2+-sensitivity and TnI phosphorylation may be unstable. In familial cardiomyopathies, both dilated and hypertrophic (DCM and HCM, a mutation in one of the proteins of the thin filament often results in the loss of the relationship (uncoupling and blunting of the lusitropic response. For familial dilated cardiomyopathy in thin filament proteins it has been proposed that this uncoupling is causative of the phenotype. Uncoupling has also been found in human heart tissue from patients with hypertrophic obstructive cardiomyopathy as a secondary effect. Recently, it has been found that Ca2+-sensitizing drugs can promote uncoupling, whilst one Ca2+-desensitising drug Epigallocatechin 3-Gallate (EGCG can reverse uncoupling.We will discuss recent findings about the role of uncoupling in the development of cardiomyopathies and the molecular mechanism of the process.

  16. Lesions of the avian pancreas.

    Science.gov (United States)

    Schmidt, Robert E; Reavill, Drury R

    2014-01-01

    Although not well described, occasional reports of avian exocrine and endocrine pancreatic disease are available. This article describes the lesions associated with common diseases of the avian pancreas reported in the literature and/or seen by the authors.

  17. The expression of uncoupling protein 2 and bone morphogenetic protein 2 in the kidney of iodine deficiency rats%碘缺乏大鼠肾脏解耦联蛋白2与骨形态发生蛋白2的表达

    Institute of Scientific and Technical Information of China (English)

    田秀标; 韩颖; 刘艳; 房辉; 阎玉琴; 林来祥

    2013-01-01

    Objective To detect the expression of uncoupling protein 2 (UCP2) and bone morphogenetic protein 2 (BMP2) in the kidney of Wistar rats,and explore the changes and distribution of them.Methods Female Wistar rats were divided into mild iodine deficiency group (MIDG group),severe iodine deficiency group(SIDG group) and normal iodine group as control (CL group) with radom number table method,and fed adaptively for one week before the experiment with ten rats in each group (everyday total iodine intaking were 5.00,1.24,10.00 μg/d,respectively).Observed the expression of UCP2 and B MP2 in isolated kidney of the experimental rats through immunohistochemistry.And the results were analyzed with related statistic methods.Results Compared with CL group,the level of free T3,total T3,total T4 in MIDG group declined (t =2.54,1.81,4.41,P < 0.05),while the level of free T4 did not decrease obviously,and the difference had no statistical significance.And the level of free T3,free T4,total T3,total T4 in SIDG group declined evidently (t =6.68,20.25,5.38,21.56,P < 0.01).Compared with MIDG group,the level ofblood hormone in SIDG group declined more obviously (t =3.19,15.45,6.93,13.48,P < 0.05).Immunohistochemistry showed that UCP2 and BMP2 expressed mainly in the cortical distal renal tubular and less in glomeruli.Compared with CL group,the expression of UCP2 and BMP2 in SIDG group and MIDG group were both decreased significantly (t =5.72,8.79,8.74,18.63,P < 0.01).Compared MIDG group,the expression in SIDG group declined more obviously (t =7.43,11.77,P < 0.01).Meanwhile,the relevant analysis revealed that the tendencies of decline of UCP2 and BMP2 were consistent with the level of free T4 (r =0.81,0.82).Conclusion The deficiency of iodine can cause hypothyroidism which can lead to the decline of UCP2 and BMP2 in kidney of rats.%目的 通过测定碘缺乏大鼠肾脏解耦联蛋白(UCP)2、骨形态发生蛋白(BMP)2的表达,探讨其表达的改变与分布.方法

  18. H5N1亚型禽流感病毒NS1蛋白研究进展%The research progress of H5N1 subtype avian influenza virus NS1 protein

    Institute of Scientific and Technical Information of China (English)

    李观强; 李国明; 张志珍

    2011-01-01

    @@ 禽流感(avian influenza ,AI) 为A型流感病毒(avian influenza virus ,AIV)引起的禽类流行性感冒.根据是否引起流感及症状的不同,通常将禽流感分为高致病性(highly pathogenic avian influenza,HPAI)、低致病性(low pathogenic avian influenza,LPAI) 和非致病性禽流感(no pathogenic avian influenza,NPAI).

  19. Exercise-induced cardioprotection: a role for eNOS uncoupling and NO metabolites.

    Science.gov (United States)

    Farah, C; Kleindienst, A; Bolea, G; Meyer, G; Gayrard, S; Geny, B; Obert, P; Cazorla, O; Tanguy, S; Reboul, Cyril

    2013-11-01

    Exercise is an efficient strategy for myocardial protection against ischemia-reperfusion (IR) injury. Although endothelial nitric oxide synthase (eNOS) is phosphorylated and activated during exercise, its role in exercise-induced cardioprotection remains unknown. This study investigated whether modulation of eNOS activation during IR could participate in the exercise-induced cardioprotection against IR injury. Hearts isolated from sedentary or exercised rats (5 weeks training) were perfused with a Langendorff apparatus and IR performed in the presence or absence of NOS inhibitors [N-nitro-L-arginine methyl ester, L-NAME or N5-(1-iminoethyl)-L-ornithine, L-NIO] or tetrahydrobiopterin (BH₄). Exercise training protected hearts against IR injury and this effect was abolished by L-NAME or by L-NIO treatment, indicating that exercise-induced cardioprotection is eNOS dependent. However, a strong reduction of eNOS phosphorylation at Ser1177 (eNOS-PSer1177) and of eNOS coupling during early reperfusion was observed in hearts from exercised rats (which showed higher eNOS-PSer1177 and eNOS dimerization at baseline) in comparison to sedentary rats. Despite eNOS uncoupling, exercised hearts had more S-nitrosylated proteins after early reperfusion and also less nitro-oxidative stress, indexed by lower malondialdehyde content and protein nitrotyrosination compared to sedentary hearts. Moreover, in exercised hearts, stabilization of eNOS dimers by BH4 treatment increased nitro-oxidative stress and then abolished the exercise-induced cardioprotection, indicating that eNOS uncoupling during IR is required for exercise-induced myocardial cardioprotection. Based on these results, we hypothesize that in the hearts of exercised animals, eNOS uncoupling associated with the improved myocardial antioxidant capacity prevents excessive NO synthesis and limits the reaction between NO and O₂·- to form peroxynitrite (ONOO⁻), which is cytotoxic.

  20. Avian influenza A (H9N2: computational molecular analysis and phylogenetic characterization of viral surface proteins isolated between 1997 and 2009 from the human population

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    Idrees Muhammad

    2010-11-01

    Full Text Available Abstract Background H9N2 avian influenza A viruses have become panzootic in Eurasia over the last decade and have caused several human infections in Asia since 1998. To study their evolution and zoonotic potential, we conducted an in silico analysis of H9N2 viruses that have infected humans between 1997 and 2009 and identified potential novel reassortments. Results A total of 22 hemagglutinin (HA and neuraminidase (NA nucleotide and deduced amino acid sequences were retrieved from the NCBI flu database. It was identified that mature peptide sequences of HA genes isolated from humans in 2009 had glutamine at position 226 (H3 of the receptor binding site, indicating a preference to bind to the human α (2-6 sialic acid receptors, which is different from previously isolated viruses and studies where the presence of leucine at the same position contributes to preference for human receptors and presence of glutamine towards avian receptors. Similarly, strains isolated in 2009 possessed new motif R-S-N-R in spite of typical R-S-S-R at the cleavage site of HA, which isn't reported before for H9N2 cases in humans. Other changes involved loss, addition, and variations in potential glycosylation sites as well as in predicted epitopes. The results of phylogenetic analysis indicated that HA and NA gene segments of H9N2 including those from current and proposed vaccine strains belong to two different Eurasian phylogenetic lineages confirming possible genetic reassortments. Conclusions These findings support the continuous evolution of avian H9N2 viruses towards human as host and are in favor of effective surveillance and better characterization studies to address this issue.

  1. Uncoupling GP1 and GP2 Expression in the Lassa Virus Glycoprotein Complex: Implications for GP1 Ectodomain Shedding

    Science.gov (United States)

    2008-12-23

    contributors Abstract Background: Sera from convalescent Lassa fever patients often contains antibodies to Lassa virus (LASV) glycoprotein 1 (GP1...uncoupled proteins will improve current diagnostics, treatment, and prevention of Lassa fever . To this end, mammalian expression systems were engineered for...stages of Lassa fever . Published: 23 December 2008 Virology Journal 2008, 5:161 doi:10.1186/1743-422X-5-161 Received: 14 December 2008 Accepted: 23

  2. Uncoupling GP1 and GP2 Expression in the Lassa Virus Glycoprotein Complex: Implications for GPI Ectodomain Shedding

    Science.gov (United States)

    2008-12-23

    contributors Abstract Background: Sera from convalescent Lassa fever patients often contains antibodies to Lassa virus (LASV) glycoprotein 1 (GP1...uncoupled proteins will improve current diagnostics, treatment, and prevention of Lassa fever . To this end, mammalian expression systems were engineered for...stages of Lassa fever . Published: 23 December 2008 Virology Journal 2008, 5:161 doi:10.1186/1743-422X-5-161 Received: 14 December 2008 Accepted: 23

  3. Administration of Poly[di(sodium carboxylatoethylphenoxy)phosphazene] (PCEP) and Avian Beta Defensin as Adjuvants in Inactivated Inclusion Body Hepatitis Virus and its Hexon Protein-Based Experimental Vaccine Formulations in Chickens.

    Science.gov (United States)

    Dar, Arshud; Tipu, Masroor; Townsend, Hugh; Potter, Andy; Gerdts, Volker; Tikoo, Suresh

    2015-12-01

    Inclusion body hepatitis (IBH) is one of the major infectious diseases adversely affecting the poultry industry of the United States and Canada. Currently, no effective and safe vaccine is available for the control of IBH virus (IBHV) infection in chickens. However, based on the excellent safety and immunogenic profiles of experimental veterinary vaccines developed with the use of new generation adjuvants, we hypothesized that characterization of vaccine formulations containing inactivated IBHV or its capsid protein hexon as antigens, along with poly[di(sodium carboxylatoethylphenoxy)phosphazene] (PCEP) and avian beta defensin 2 (ABD2) as vaccine adjuvants, will be helpful in development of an effective and safe vaccine formulation for IBH. Our data demonstrated that experimental administration of vaccine formulations containing inactivated IBHV and a mixture of PCEP with or without ABD2 as an adjuvant induced significantly higher antibody responses compared with other vaccine formulations, while hexon protein-based vaccine formulations showed relatively lower levels of antibody responses. Thus, a vaccine formulation containing inactivated IBHV with PCEP or a mixture of PCEP and ABD2 (with a reduced dosage of PCEP) as an adjuvant may serve as a potential vaccine candidate. However, in order to overcome the risks associated with whole virus inactivated vaccines, characterization of additional viral capsid proteins, including fiber protein and penton of IBHV along with hexon protein in combination with more new generation adjuvants, will be helpful in further improvements of vaccines against IBHV infection.

  4. Homosubtypic and heterosubtypic antibodies against highly pathogenic avian influenza H5N1 recombinant proteins in H5N1 survivors and non-H5N1 subjects.

    Science.gov (United States)

    Noisumdaeng, Pirom; Pooruk, Phisanu; Prasertsopon, Jarunee; Assanasen, Susan; Kitphati, Rungrueng; Auewarakul, Prasert; Puthavathana, Pilaipan

    2014-04-01

    Six recombinant vaccinia viruses containing HA, NA, NP, M or NS gene insert derived from a highly pathogenic avian influenza H5N1 virus, and the recombinant vaccinia virus harboring plasmid backbone as the virus control were constructed. The recombinant proteins were characterized for their expression and subcellular locations in TK(-) cells. Antibodies to the five recombinant proteins were detected in all 13 sequential serum samples collected from four H5N1 survivors during four years of follow-up; and those directed to rVac-H5 HA and rVac-NA proteins were found in higher titers than those directed to the internal proteins as revealed by indirect immunofluorescence assay. Although all 28 non-H5N1 subjects had no neutralizing antibodies against H5N1 virus, they did have cross-reactive antibodies to those five recombinant proteins. A significant increase in cross-reactive antibody titer to rVac-H5 HA and rVac-NA was found in paired blood samples from patients infected with the 2009 pandemic virus.

  5. Quantum Zeno Effect Underpinning the Radical-Ion-Pair Mechanism of Avian Magnetoreception

    OpenAIRE

    Kominis, I. K.

    2008-01-01

    The intricate biochemical processes underlying avian magnetoreception, the sensory ability of migratory birds to navigate using earths magnetic field, have been narrowed down to spin-dependent recombination of radical-ion pairs to be found in avian species retinal proteins. The avian magnetic field detection is governed by the interplay between magnetic interactions of the radicals unpaired electrons and the radicals recombination dynamics. Critical to this mechanism is the long lifetime of t...

  6. Association between mitochondrial uncoupling protein 2 gene promoter -866G>A polymorphism and ischemic stroke in diabetic patients%线粒体解耦联蛋白2基因启动子-866G>A多态性与2型糖尿病缺血性脑卒中的相关性研究

    Institute of Scientific and Technical Information of China (English)

    顾兵; 仇金荣; 朱倩; 张璐; 柴怡

    2012-01-01

    目的 探讨线粒体解耦联蛋白2 (UcP-2)基因启动子-866G>A变异与2型糖尿病患者合并缺血性脑卒中(IS)发生风险的相关性.方法 对844例2型糖尿病患者(伴IS组404例,不伴IS组440例)进行为期4年的前瞻性队列研究,提取所有受试者的基因组全血DNA,用TaqMan MGB探针对UCP-2基因启动子-866G>A进行基因分型,分析不同基因型与2型糖尿病合并IS发生风险的关联性.结果 无论在基线时还是在4年随访过程中,携带A等位基因的2型糖尿病女性患者IS发生风险明显高于GG野生型(P<0.05),但男性患者3种基因型之间差异无统计学意义.结论 UCP-2基因启动子-866G>A变异增加中国女性2型糖尿病并发IS的风险.%Objective To investigate the association of uncoupling protein 2 ( UCP-2 ) gene promoter -866G>A polymorphism and ischemic stroke in diabetic patients.Methods A total of 844 type 2 diabetic patients including 404 cases with ischemic stroke and 440 cases without ischemic stroke were selected for the 4 year prospective study,Genomic DNA was extracted from the whole blood samples of subjects,UCP-2 gene promoter -866G > A polymorphism was detected by TaqMan MGB probe method,and then the genotype and allele gene frequencies were compared.Results The risk of ischemic stroke in type 2 diabetic female patients with AA+GA genotypes of UCP-2 was higher than that with GG genotype (P<0.05),but there was no difference among male patients with three genotypes.Conclusions UCP-2 gene promoter -866G > A polymorphism increases the risk of ischemic stroke in Chinese diabetic women.

  7. Analysis of-3826A/G polymorphism in the promoter of the uncoupling protein-1 gene in Chinese non-obese and obese populations%正常中国人及肥胖患者解偶联蛋白1基因-3826 A/G多态性的研究

    Institute of Scientific and Technical Information of China (English)

    沈哲旎; 王晓苏; 白怀; 范平; 刘瑞; 刘宇; 刘秉文

    2009-01-01

    Objective To investigate the-3826A/G polymorphism in the promoter of the uncoupling protein-1(UCP1) gene and its relations to obesity in Chinese population. Methods Three hundred and eighty-four subjects (257 non-obese and 127 obese individuals) from a population of Chinese Han nationality in Chengdu area were studied using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs). Serum lipids were measured by enzymatic kits and apolipoproteins AⅠ, AⅡ, B100, CⅡ,CⅢ and E were measured by the RID kits. Results The frequencies of A and G alleles at-3826A/G site in obese and non-obese groups were 0.508 and 0. 492, and 0. 467 and 0. 533, respectively. It showed no significant difference in allele frequencies between non-obese and obese groups (P>0.05). In the obese group, subjects with genotype GG had higher serum apo B100 concentrations, and those with genotype AG had higher apo CⅡ and apo CⅢ levels, than those with genotype AA, respectively (P0.05).-3826 A/G位点在肥胖组GG基因型携带者血清载脂蛋白(apolipoprotein,apo)B100水平高于AA基因型者(P<0.05),AG型者apo CⅡ、apo CⅢ水平均分别高于AA基因型携带者(P<0.05).在非肥胖男性亚组,GG纯合基因型携带者血清高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)及apoAⅠ水平显著低于AA基因型携带者(P<0.05);AG型者apoAⅡ水平低于AA基因型者.在肥胖男性亚组,GG基因型携带者血清apo B100水平较AA型者显著升高(P<0.05);在肥胖女性亚组,GG基因型者apo AⅠ水平低于AG型者,AG型者apoCⅡ、apoCⅢ水平均分别高于AA型者(P<0.05).结论 UCP1基因-3826A/G多态性与成都地区中国汉族人肥胖无关联,但与血清HDL-C及apoAⅠ、apoB100等载脂蛋白水平含量有关,且具有性别差异.

  8. Effect of propofol on rat forebrain ischemia reperfusion induced mitochondrial damage and uncoupling protein 2 expression%丙泊酚对大鼠前脑缺血/再灌注诱导线粒体损伤及解耦联蛋白2表达的影响

    Institute of Scientific and Technical Information of China (English)

    戚思华; 王晓东; 李军; 王毅; 李文志

    2010-01-01

    Objective To investigate the effect of propofol on rat forebrain ischemia reperfusion induced mitochondrial damage and uncoupling protein 2 (UCP 2)expression. Methods 45 male Wistar rats weighing 250 g-300 g were randomly divided into three groups (n= 15):control group (C) ;ischemia reperfusion group (I/R) ;propofol group (P). Forebrain cerebral ischemia reperfusion was produced by 2-vessel occlusion method. Bilateral carotid arteries were released after 10 min cerebral ischemia. Normal saline 1 mg/kg and propofol 1 mg/kg were separately injected into the lateral cerebral ventricle by micro syringe in group I/R and group P. The animals were decapitated at the end of 24 h reperfusion and the hippocampal were separated.The mitochondria of hippocampal in each group were isolated. The mitochondria in three groups were incubated by CaCl2for 5 min at 37℃. Morphological changes of mitoehondria were observed by using electron microscopy (n=3). Mitochondrial permeability transition pore (MPTP)opening were detected by ultravioletvisible absorption spectroscopy(n=6). The expression of UCP2 in each group were determined by western blotting method (n=6). Results Group C showed normal mitochondrial ultrastructure;Significant mitochondrial swelling, disrupted cristae and membrane rupture were showed in group I/R;Morphological changes of mitechondria in group P were between group C and group I/R. Absorbance values of mitochondrial decreased in group C, group I/R and group P. Compared with group C, absorbance values of mitochondrial were lower in group I/R and group P (P<0.05). The decrease of absorbance values of mitochondria was reduced in group P(0.017 ± 0.007)compared with that in group I/R (0.028 ± 0.007)(P<0.05). The expression of UCP2 in hippocampus was significantly up-regnlated in group I/R (0.88 ± 0.14) and group P (1.32 + 0. 10)(P<0.05). Compared with group I/R, the expression of UCP2 were higher in group P (P<0.05). Conclusion Propofol improve mitochondrial

  9. Avaliação do perfil eletroforético das proteínas séricas em matrizes pesadas (Gallus Gallus Domesticus da linhagem Avian Farm Serum protein electrophoresis evaluation in adult broiler breeders (Gallus Gallus Domesticus of the Avian Farm strain

    Directory of Open Access Journals (Sweden)

    MY Hasegawa

    2002-12-01

    Full Text Available O objetivo do presente trabalho foi determinar o perfil eletroforético das proteínas séricas em matrizes pesadas (Gallus Gallus Domesticus sadias, da linhagem Avian Farm. Foram utilizadas 15 matrizes com 63 semanas de idade, provenientes do município de Conchas, São Paulo. Utilizou-se o método de biureto para a obtenção dos valores da proteína sérica total e a separação das frações protéicas pela técnica da eletroforese em gel de agarose, e a leitura do filme realizada por densitometria em 520nM. Obteve-se um total de sete frações, sendo que a beta1 - globulina e beta2 - globulina encontradas não foram citadas pelos autores consultados na literatura. A fração pré-albumina foi identificada em apenas seis das 15 amostras examinadas. Observou-se em cinco matrizes a divisão da gama - globulina em duas frações, denominadas gama - 1 e gama - 2, de acordo com suas mobilidades eletroforéticas. A relação albumina/globulina (A/G encontrada corrobora com os autores citados, demonstrando que esta diminui com o aumento da idade.This work is aimed to determine the profile of electrophoretic serum protein in healthy adult broiler breeders (Gallus gallus domesticus of the Avian farm strain. Fifteen breeders aging 63 weeks from Conchas, city located in the State of São Paulo, were assessed. The biuret method was used to obtain the total serum protein values and protein fractions separation through electrophoresis technique in agarose gel, and film reading through densitometry in 520nM. Seven fractions were obtained, whereas, beta1 - globulin and beta2 - globulin were not cited by the authors in the textbooks checked. The prealbumin fraction was identified only in six out of 15 samples analyzed. In five breeders, it was observed the division of gamma - globulin into two fractions named gamma - 1 and gamma - 2, according to the electrophoretic mobilities. The relation albumin/globulin (A/G found in the experiment agrees with the other

  10. 脾阳虚证大鼠棕色脂肪组织和解偶联蛋白1关联性的研究%Experimental Research on Relationship between Brown Adipose Tissue and Uncoupling Protein 1 in Rats with Spleen Yang Deficiency Syndrome

    Institute of Scientific and Technical Information of China (English)

    吴云起; 唐汉庆; 吴翠松; 劳传君; 庞广福

    2011-01-01

    Objective:To explore the relationship and mechanism about brown adipose tissue, uncoupling protein 1 (UCP1), high fatty diet and Chinese herbal decoction during the process of energy metabolism. Method: Rats were divided into four groups (n = 16 each). ①Control group was fed with ordinary food, also ig given 0.9% NaCl 10 g·kg-1 during d70-d98 daily. The ability of ISO deducing heat generating was tested on d98, and the curve of the rectal temperature was monitored, and the peak within two hours and the area under the curve were determined.②Yang deficiency syndrome group: the same procedure was carried out as the control group except removing brown adipose tissue on d42. ③Spleen yang deficiency syndrome group: the same procedure was carried out as the yang deficiency syndrome group except of being fed with high fatty diet (including 83% ordinary diet,15% triglycerides,2% cholesterol) from d49 to d98 and placed in 19 ℃ environment every other day. ④Chinese herbal decoction group: the same procedure was carried out as the spleen yang deficiency syndrome group except giving 4 g·kg-1 ig daily with Aconitum Lizhong decoction and the body weight was measured every week. The index of the weight growth rate, the peak of temperature and the area under the temperature curve were all investigated. The content of UCP1 was determined by using brown adipose tissue. Result: ①Weight growth rate: compared with yang deficiency syndrome group, weight growth rate in spleen yang deficiency syndrome group was lower( P < 0. 01 ); compared with spleen yang deficiency syndrome group, weight growth rate was higher in the Chinese herbal decoction group (P < 0.01 ). ②Temperature: compared with yang deficiency syndrome group, the temperature peak of heat generating in spleen yang deficiency syndrome group was higher(P < 0.01 ); compared with spleen yang deficiency syndrome group, temperature peak of heat generating was higher in Chinese herbal decoction group ( P

  11. Uncoupler resistance in E. coli Tuv and Cuv is due to the exclusion of uncoupler by the outer membrane

    DEFF Research Database (Denmark)

    Haworth, Robert S.; Jensen, Peter Ruhdal; Michelsen, Ole;

    1990-01-01

    The uncoupler resistant bacterial strains E. coli Tuv and Cuv share the high deoxycholate sensitivity of the parent strain, Doc S. However, both Tuv and Cuv show greater resistance than Doc S to other detergents. Measurement of the periplasmic volume indicates that the outer membrane of Doc S...

  12. Avian influenza control strategies

    Science.gov (United States)

    Control strategies for avian influenza in poultry vary depending on whether the goal is prevention, management, or eradication. Components used in control programs include: 1) education which includes communication, public awareness, and behavioral change, 2) changes to production and marketing sys...

  13. Avian influenza virus

    Science.gov (United States)

    Avian influenza (AI) is caused by type A influenza virus, a member of the Orthomyxoviridae family. AI viruses are serologically categorized into 16 hemagglutinin (H1-H16) and 9 neuraminidase (N1-N9) subtypes. All subtypes have been identified in birds. Infections by AI viruses have been reported in ...

  14. Uncoupling effect of fatty acids in halo- and alkalotolerant bacterium Bacillus pseudofirmus FTU.

    Science.gov (United States)

    Popova, I V; Bodrova, M E; Mokhova, E N; Muntyan, M S

    2004-10-01

    Natural uncouplers of oxidative phosphorylation, long-chain non-esterified fatty acids, cause uncoupling in the alkalo- and halotolerant bacterium Bacillus pseudofirmus FTU. The uncoupling effect in the bacterial cells was manifested as decrease of membrane potential and increase of respiratory activity. The membrane potential decrease was detected only in bacterial cells exhausted by their endogenous substrates. In proteoliposomes containing reconstituted bacterial cytochrome c oxidase, fatty acids caused a "mild" uncoupling effect by reducing membrane potential only at low rate of membrane potential generation. "Free respiration" induced by the "mild" uncouplers, the fatty acids, can be considered as possible mechanism responsible for adaptation of the bacteria to a constantly changed environment.

  15. 禽流感病毒非结构蛋白NS1在感染和免疫中的作用%Roles of Avian Influenza Virus Non-structural 1 Protein in Infection and Immunity

    Institute of Scientific and Technical Information of China (English)

    路冰; 陈化兰; 包红梅; 王秀荣; 张秀英

    2011-01-01

    NS1蛋白(non-structural 1 protein)是目前发现的唯一的禽流感病毒非结构蛋白,它是一种多功能调节蛋白,对病毒的感染、复制和毒力具有重要的作用;该蛋白主要通过颉颃宿主干扰素及肿瘤坏死因子,抑制宿主蛋白的合成及调控感染细胞的凋亡来实现其作用.作者综述了NS1蛋白生物学特性及其在感染和免疫中作用的研究进展,为进一步研究NS1蛋白的特性及临床应用提供了参考.%NS1 (non-structural 1) protein is the unique non-structural protein found in avian influenza virus.It is a kind of multifunctional regulatory protein which plays a vital role in virus infection,replication and virulence.Its main function is antagonizing the host interferon and tumor necrosis factor,production inhibiting host protein synthesis as well as regulating apoptosis of infected cells.This paper reviews the latest research on the biological characteristics of NS1 protein and its role in infection and immunity which provide a reference for further research on NS1 protein's characteristics and clinical application.

  16. Within brown-fat cells, UCP1-mediated fatty acid-induced uncoupling is independent of fatty acid metabolism.

    Science.gov (United States)

    Shabalina, Irina G; Backlund, Emma C; Bar-Tana, Jacob; Cannon, Barbara; Nedergaard, Jan

    2008-01-01

    In the present investigation, we have utilized the availability of UCP1(-/-) mice to examine a wide range of previously proposed lipid activators of Uncoupling Protein 1 (UCP1) in its native environment, i.e. in the brown-fat cells. A non-metabolizable fatty acid analogue, beta,beta cent-methyl-substituted hexadecane alpha,omega-dicarboxylic acid (Medica-16) is a potent UCP1 (re)activator in brown-fat cells, despite its bipolar structure. All-trans-retinoic acid activates UCP1 within cells, whereas beta-carotene only does so after metabolism. The UCP1-dependent effects of fatty acids are positively correlated with their chain length. Medium-chain fatty acids are potent UCP1 activators in cells, despite their lack of protonophoric properties in mitochondrial membranes. Thus, neither the ability to be metabolized nor an innate uncoupling/protonophoric ability is a necessary property of UCP1 activators within brown-fat cells.

  17. eNOS uncoupling in the cerebellum after BBB disruption by exposure to Phoneutria nigriventer spider venom.

    Science.gov (United States)

    Soares, Edilene Siqueira; Mendonça, Monique Culturato Padilha; da Cruz-Höfling, Maria Alice

    2015-09-15

    Numerous studies have shown that the venom of Phoneutria nigriventer (PNV) armed-spider causes excitotoxic signals and blood-brain barrier breakdown (BBBb) in rats. Nitric oxide (NO) is a signaling molecule which has a role in endothelium homeostasis and vascular health. The present study investigated the relevance of endothelial NO synthase (eNOS) uncoupling to clinical neurotoxic evolution induced by PNV. eNOS immunoblotting of cerebellum lysates processed through low-temperature SDS-PAGE revealed significant increased monomerization of the enzyme at critical periods of severe envenoming (1-2 h), whereas eNOS dimerization reversal paralleled to amelioration of animals condition (5-72 h). Moreover, eNOS uncoupling was accompanied by increased expression in calcium-sensing calmodulin protein and calcium-binding calbindin-D28 protein in cerebellar neurons. It is known that greater eNOS monomers than dimers implies the inability of eNOS to produce NO leading to superoxide production and endothelial/vascular barrier dysfunction. We suggest that transient eNOS deactivation and disturbances in calcium handling reduce NO production and enhance production of free radicals thus contributing to endothelial dysfunction in the cerebellum of envenomed rats. In addition, eNOS uncoupling compromises the enzyme capacity to respond to shear stress contributing to perivascular edema and it is one of the mechanisms involved in the BBBb promoted by PNV.

  18. Nox2-dependent glutathionylation of endothelial NOS leads to uncoupled superoxide production and endothelial barrier dysfunction in acute lung injury.

    Science.gov (United States)

    Wu, Feng; Szczepaniak, William S; Shiva, Sruti; Liu, Huanbo; Wang, Yinna; Wang, Ling; Wang, Ying; Kelley, Eric E; Chen, Alex F; Gladwin, Mark T; McVerry, Bryan J

    2014-12-15

    Microvascular barrier integrity is dependent on bioavailable nitric oxide (NO) produced locally by endothelial NO synthase (eNOS). Under conditions of limited substrate or cofactor availability or by enzymatic modification, eNOS may become uncoupled, producing superoxide in lieu of NO. This study was designed to investigate how eNOS-dependent superoxide production contributes to endothelial barrier dysfunction in inflammatory lung injury and its regulation. C57BL/6J mice were challenged with intratracheal LPS. Bronchoalveolar lavage fluid was analyzed for protein accumulation, and lung tissue homogenate was assayed for endothelial NOS content and function. Human lung microvascular endothelial cell (HLMVEC) monolayers were exposed to LPS in vitro, and barrier integrity and superoxide production were measured. Biopterin species were quantified, and coimmunoprecipitation (Co-IP) assays were performed to identify protein interactions with eNOS that putatively drive uncoupling. Mice exposed to LPS demonstrated eNOS-dependent increased alveolar permeability without evidence for altered canonical NO signaling. LPS-induced superoxide production and permeability in HLMVEC were inhibited by the NOS inhibitor nitro-l-arginine methyl ester, eNOS-targeted siRNA, the eNOS cofactor tetrahydrobiopterin, and superoxide dismutase. Co-IP indicated that LPS stimulated the association of eNOS with NADPH oxidase 2 (Nox2), which correlated with augmented eNOS S-glutathionylation both in vitro and in vivo. In vitro, Nox2-specific inhibition prevented LPS-induced eNOS modification and increases in both superoxide production and permeability. These data indicate that eNOS uncoupling contributes to superoxide production and barrier dysfunction in the lung microvasculature after exposure to LPS. Furthermore, the results implicate Nox2-mediated eNOS-S-glutathionylation as a mechanism underlying LPS-induced eNOS uncoupling in the lung microvasculature.

  19. Mechanism of fatty-acid-dependent UCP1 uncoupling in brown fat mitochondria.

    Science.gov (United States)

    Fedorenko, Andriy; Lishko, Polina V; Kirichok, Yuriy

    2012-10-12

    Mitochondrial uncoupling protein 1 (UCP1) is responsible for nonshivering thermogenesis in brown adipose tissue (BAT). Upon activation by long-chain fatty acids (LCFAs), UCP1 increases the conductance of the inner mitochondrial membrane (IMM) to make BAT mitochondria generate heat rather than ATP. Despite being a member of the family of mitochondrial anion carriers (SLC25), UCP1 is believed to transport H(+) by an unusual mechanism that has long remained unresolved. Here, we achieved direct patch-clamp measurements of UCP1 currents from the IMM of BAT mitochondria. We show that UCP1 is an LCFA anion/H(+) symporter. However, the LCFA anions cannot dissociate from UCP1 due to hydrophobic interactions established by their hydrophobic tails, and UCP1 effectively operates as an H(+) carrier activated by LCFA. A similar LCFA-dependent mechanism of transmembrane H(+) transport may be employed by other SLC25 members and be responsible for mitochondrial uncoupling and regulation of metabolic efficiency in various tissues.

  20. Arsenate uncoupling of oxidative phosphorylation in isolated plant mitochondria

    Energy Technology Data Exchange (ETDEWEB)

    Wickes, W.A.; Wiskich, J.T.

    1976-01-01

    The uncoupling by arsenate of beetroot and cauliflower bud mitochondria showed the following characteristics: arsenate stimulation of respiration above the rate found with phosphate; inhibition of arsenate-stimulated respiration by phosphate; enhancement of arsenate-stimulated respiration by ADP; only partial prevention of this ADP-enhanced respiration by atractyloside; inhibition by oligomycin of the arsenate-stimulated respiration back to the phosphate rate; and the absence of any stimulatory effect of ADP in the presence of oligomycin. These results are qualitatively analogous to those reported for arsenate uncoupling in rat liver mitochondria. Arsenate stimulated malate oxidation, presumably by stimulating malate entry, in both beetroot and cauliflower bud mitochondria; however, high rates of oxidation, and presumably entry, were only sustained with arsenate in beetroot mitochondria. NADH was oxidized rapidly in cauliflower bud mitochondria in the presence of arsenate, showing that arsenate did not inhibit electron transfer processes.

  1. Avian cytokines in health and disease

    Directory of Open Access Journals (Sweden)

    P Wigley

    2003-04-01

    Full Text Available Cytokines are proteins secreted by cells that play an important role in the activation and regulation of other cells and tissues during inflammation and immune responses. Although well described in several mammalian species, the role of cytokines and other related proteins is poorly understood in avian species. Recent advances in avian genetics and immunology have begun to allow the exploration of cytokines in health and disease. Cytokines may be classified in a number of ways, but may be conveniently arranged into four broad groups on the basis of their function. Proinflammatory cytokines such as interleukin-6 and interleukin-1beta play a role in mediating inflammation during disease or injury. Th1 cytokines, including interleukin-12 and interferon-gamma, are involved in the induction of cell-mediated immunity, whereas Th2 cytokines such as interleukin-4 are involved in the induction of humoral immunity. The final group Th3 or Tr cytokines play a role in regulation of immunity. The role of various cytokines in infectious and non-infectious diseases of chickens and turkeys is now being investigated. Although there are only a few reliable ELISAs or bioassays developed for avian cytokines, the use of molecular techniques, and in particular quantitative RT-PCR (Taqman has allowed investigation of cytokine responses in a number of diseases including salmonellosis, coccidiosis and autoimmune thyroiditis. In addition the use of recombinant cytokines as therapeutic agents or as vaccine adjuvants is now being explored.

  2. Uncoupling of Vascular Nitric Oxide Synthase Caused by Intermittent Hypoxia

    Directory of Open Access Journals (Sweden)

    Mohammad Badran

    2016-01-01

    Full Text Available Objective. Obstructive sleep apnea (OSA, characterized by chronic intermittent hypoxia (CIH, is often present in diabetic (DB patients. Both conditions are associated with endothelial dysfunction and cardiovascular disease. We hypothesized that diabetic endothelial dysfunction is further compromised by CIH. Methods. Adult male diabetic (BKS.Cg-Dock7m +/+ Leprdb/J (db/db mice (10 weeks old and their heterozygote littermates were subjected to CIH or intermittent air (IA for 8 weeks. Mice were separated into 4 groups: IA (intermittent air nondiabetic, IH (intermittent hypoxia nondiabetic, IADB (intermittent air diabetic, and IHDB (intermittent hypoxia diabetic groups. Endothelium-dependent and endothelium-independent relaxation and modulation by basal nitric oxide (NO were analyzed using wire myograph. Plasma 8-isoprostane, interleukin-6 (IL-6, and asymmetric dimethylarginine (ADMA were measured using ELISA. Uncoupling of eNOS was measured using dihydroethidium (DHE staining. Results. Endothelium-dependent vasodilation and basal NO production were significantly impaired in the IH and IADB group compared to IA group but was more pronounced in IHDB group. Levels of 8-isoprostane, IL-6, ADMA, and eNOS uncoupling were ≈2-fold higher in IH and IADB groups and were further increased in the IHDB group. Conclusion. Endothelial dysfunction is more pronounced in diabetic mice subjected to CIH compared to diabetic or CIH mice alone. Oxidative stress, ADMA, and eNOS uncoupling were exacerbated by CIH in diabetic mice.

  3. BIRD FLU (AVIAN INFLUENZA)

    OpenAIRE

    Ali ACAR; Bulent BESIRBELLIOÐLU

    2005-01-01

    Avian influenza (bird flu) is a contagious disease of animals caused by influenza A viruses. These flu viruses occur naturally among birds. Actually, humans are not infected by bird flu viruses.. However, during an outbreak of bird flu among poultry, there is a possible risk to people who have contact infect birds or surface that have been contaminated with excreations from infected birds. Symptoms of bird flu in humans have ranged from typical flu-like symptoms to eye infections, pneumonia, ...

  4. Grid attacks avian flu

    CERN Multimedia

    2006-01-01

    During April, a collaboration of Asian and European laboratories analysed 300,000 possible drug components against the avian flu virus H5N1 using the EGEE Grid infrastructure. Schematic presentation of the avian flu virus.The distribution of the EGEE sites in the world on which the avian flu scan was performed. The goal was to find potential compounds that can inhibit the activities of an enzyme on the surface of the influenza virus, the so-called neuraminidase, subtype N1. Using the Grid to identify the most promising leads for biological tests could speed up the development process for drugs against the influenza virus. Co-ordinated by CERN and funded by the European Commission, the EGEE project (Enabling Grids for E-sciencE) aims to set up a worldwide grid infrastructure for science. The challenge of the in silico drug discovery application is to identify those molecules which can dock on the active sites of the virus in order to inhibit its action. To study the impact of small scale mutations on drug r...

  5. Association between uncoupling protein 2 gene polymorphism and macroangiopathy in diabetes mellitus%解偶联蛋白2基因多态性与糖尿病并发大血管病变的相关性分析

    Institute of Scientific and Technical Information of China (English)

    孙艳荪; 张宇光; 苏本利; 李昌臣

    2011-01-01

    Objective To investigate the effects of uncoupling protein 2 (UCP-2) gene a 45 bp insertion/deletion (Ins/Del) polymorphism in the 3'-untranslated (3'-UTR) of its exon 8 on macroangiopathy in diabetes mellitus.Methods A total of 182 patients were selected,80 cases with macroangiopathy( A group), 102 cases without macroangiopathy(B group) ,UCP-2 gene polymorphism was confirmed by electrophoresis after PCR with 3% agarose,then compared genotype and allele gene frequency. Results The 3'-UTR Ins/Del polymorphism of UCP-2 gene in A group ( II:6. 25% 、ID: 18. 75% 、DD:75.00% ) and B group( II:9. 80% 、ID:23.53%、 DD: 66. 67% ) had no significant difference ( P > 0. 05 ), and there was also no difference of alleles frequencies in two groups ( I: 15.63 %、 D: 84. 37 % )and(I:21.57% 、D:78.43%)(P >0.05). Conclusion No relationship of the 3'-UTR a 45bp Ina/Del polymorphism in exon 8 of the UCP-2 gene was found with macroangiopathy in diabetes mellitus.%目的 探讨解偶联蛋白2(UCP-2)基因8号外显子3'-末端(3'-UTR)45bp插入/缺失多态性与糖尿病并发大血管病变的关系.方法 182例糖尿病患者中并发大血管病变患者80例(A组)、无大血管病变患者102例(B组),检测两组基因组DNA,用聚合酶链反应(PCR)方法对UCP-2基因进行扩增,记录各组UCP-2等位基因及基因型频率并进行比较.结果 UCP-2基因3'-UTR 45bp插入/缺失多态性在A组和B组中基因型分布分别为(Ⅱ:6.25%、ID:18.75%、DD:75.00%)和(Ⅱ:9.80%、ID:23.53%、DD:66.67%),两组差异均无统计学意义(均P>0.05),两组等位基因频率分别为(Ⅰ:15.63%、D:84.37%)和(Ⅰ:21.57%、D:78.43%),两组间差异均无统计学意义(均P>0.05).结论 UCP-2基因8号外显子3'-末端(3'-UTR)45bp插入/缺失多态性与糖尿病并发大血管病变无相关性.

  6. 己酮可可碱对非酒精性脂肪肝大鼠肝组织线粒体解耦联蛋白表达的影响%Effects of pentoxifylline on expression of uncoupling protein 2 of hepatic mitochondria in rats with nonalcoholic fatty liver

    Institute of Scientific and Technical Information of China (English)

    徐丽姝; 张瑛华

    2010-01-01

    目的 探讨己酮可可碱(PTX)对非酒精性脂肪肝性肝病(NAFLD)大鼠线粒体解耦联蛋白(UCP)-2 mRNA表达的影响. 方法 SD大鼠60只,正常喂养1周后随机分为3组,每组20只.其中对照组20只,以普通饲料喂养,实验组和干预组各20只,以高脂饲料喂养,干预组高脂饮食4周后,在饮水中加用已酮可可碱(PTX)(100 mg·Kg-1·d-1),于第24周处死,采用反转录聚合酶链反应(RT-PCR)技术检测肝脏UCP-2mRNA的表达. 结果 对照组大鼠肝脏UCP2mRNA呈微量表达(1.2±0.1),实验组大鼠肝脏UCP2mRNA呈强表达(4.0±0.3),干预组大鼠肝脏UCP2mRNA呈弱表达(3.0±0.2),3组间UCP-2mRNA表达差异有统计学意义(F=160.67,P<0.01). 结论 己酮可可碱可下调NAFLD大鼠肝细胞UCP-2mRNA的表达.%Objective To investigate the effects of pentoxifylline (PTX) on the expression of uncoupling protein 2 (UCP-2) of hepatic mitochondria in rats with nonalcoholic fatty liver disease (NAFLD). Methods Sixty SD rats were randomly divided into 3 groups (each n = 20): normal control group, experiment group and treatment group. The rats in normal control group were given a normal feed. The rats in experiment and treatment groups were given fat-rich feed. Furthermore, the rats in treatment group were given PTX after 4 weeks in fat-rich diet feeding. The expression of UCP-2 in the liver was detected by immunohistochemistry and semi-quantitative RT-PCR. Results The hepatic expression of UCP2 mRNA was higher in experiment group (4.0±0.3) than in normal control group (1.2±0.1). The hepatic expression of UCP2 mRNA was higher in treatment group (3.0±0.2) than in normal control group, but the hepatic expression of UCP2 mRNA was lower in treatment group than in experiment group (F = 160. 67, P< 0. 01). Conclusions The UCP-2 mRNA is expressed in livers of NAFLD, pentoxifylline plays an important role in the reduction of expression of UCP-2 mRNA in lives of NAFLD.

  7. Relationship between uncoupling protein 2 gene promoter-866 G/A polymorphism and Chinese diabetic nephropathy%解耦联蛋白2基因启动子-866G/A多态性与中国人糖尿病肾病的关系

    Institute of Scientific and Technical Information of China (English)

    孙艳荪; 龙霞; 史嵘; 张帆; 邓远飞; 于洁

    2011-01-01

    Objective To investigate the relationship between uncoupling protein 2 (UCP-2) gene promoter -866 G/A polymorphism and Chinese diabetic nephropathy (DN). Methods A total of 182 patients with type 2 diabetic mellitus were selected and divided into DN group (90 cases with DN ) and NCD group (92 cases without DN ). Genomic DNA was detected, and UCP-2 genotype and allele gene frequency was confirmed by polymerase chain reaction-restrictive fragment length polymorphism, then compared.Results The genotype frequency of UCP-2 gene promoter-866 G/A polymorphism was distributed in DN group[GG 14.44%( 13/90),GA 31.11%(28/90),AA 54.44%(49/90)],and distributed in NCD group[GG 29.35% ( 27/92 ), GA 32.61% ( 30/92 ), AA 38.04% ( 35/92 )], and there was significant difference between two groups ( χ2 = 7.28 , P < 0.05 ). There was also significant difference in allele gene frequency between DN group and NCD group[G 45.65% (84/184) vs. 30.00% (54/180),A 54.35% (100/184) vs. 70.00% (126/180)]( χ2 = 9.47, P < 0.05 ). Conclusion There is correlation between the UCP-2 gene promoter-866 G/A polymorphism and Chinese DN, and the incidence of DN with A/A genotype is increased.%目的 探讨解耦联蛋白2(UCP-2)基因启动子-866G/A多态性与中国人糖尿病肾病(DN)的关系.方法 182例2型糖尿病患者,其中并发DN90例(DN组),无肾病92例(NCD组),检测所有患者的基因组DNA,采用聚合酶链反应-限制性长度多态性方法,记录UCP-2等位基因及基因型频率并进行比较.结果 UCP-2基因启动子-866 G/A多态性在DN组中基因型频率分布,GG 14.44%(13/90),GA 31.11%(28/90),AA 54.44%(49/90),在NCD组中基因型频率分布,GG29.35%(27/92),GA 32.61%(30/92),AA 38.04%(35/92),两组比较差异有统计学意义(χ2=7.28,P< 0.05);NCD组UCP-2等位基因频率分布,G 45.65%(84/184),A 54.35%(100/184),DN组UCP-2等位基因频率分布,G 30.00%(54/180),A 70.00%(126/180),两组比较差异有统计学意义(χ2=9.47,P<0.05).结论 UCP-2

  8. 棕榈酸对模拟高原低氧大鼠离体脑线粒体解耦联蛋白活性及质子漏的影响%Effects of palmitic acid on activity of uncoupling proteins and proton leak in in vitro cerebral mitochondria from the rats exposed to simulated high altitude nypoxia

    Institute of Scientific and Technical Information of China (English)

    徐瑜; 柳君泽; 夏琛

    2008-01-01

    本文旨在通过观察棕榈酸对模拟高原低氧大鼠离体脑线粒体解耦联蛋白(uncoupling proteins,UCPs)活性的影响及脑线粒体质子漏与膜电位的改变,探讨UCPs在介导游离脂肪酸对低氧时线粒体氧化磷酸化功能改变中的作用.将SpragueDawley大鼠随机分为对照组、急性低氧组和慢性低氧组.低氧大鼠于低压舱内模拟海拔5 000 m高原23 h/d作低氧暴露,分别连续低氧3 d和30 d.用差速密度梯度离心法提取脑线粒体,[3H-GTP法测定UCPs含量与活性,TPMP+电极与Clark氧电极结合法测量线粒体质子漏,罗丹明123荧光法测定线粒体膜电位.结果显示,低氧使脑线粒体内UCPs含量与活性升高、质子漏增加、线粒体膜电位降低;同时,低氧暴露降低脑线粒体对棕榈酸的反应性,UCPs活性的改变率低于对照组,且线粒体UCPs含量、质子漏、膜电位变化率亦出现相同趋势.线粒体质子漏与反映UCPs活性的Kd值呈线性负相关(P<0.01 r=-0.906),与反映UCPs含量的Bmax呈线性正相关(P<0.01,r=0.856),与膜电位呈线性负相关(P<0.01,r=-0.880).以上结果提示,低氧导致的脑线粒体质子漏增加及膜电位降低与线粒体内UCPs活性升高有关,同时低氧暴露能降低脑线粒体对棕榈酸的反应性,提示在高原低氧环境下,游离脂肪酸升高在维持线粒体能量代谢中起着自身保护和调节机制.

  9. Preparation and Characterization of the Monoclonal Antibodies Against p27 Protein of Avian Leukosis Virus%禽白血病病毒 p27蛋白单克隆抗体的制备及鉴定

    Institute of Scientific and Technical Information of China (English)

    谢华丽; 邵华斌; 杨峻; 程国富; 胡薛英; 谷长勤; 张万坡; 罗青平

    2013-01-01

    The purified recombinant protein p27 of avian leukosis virus as an immunogen was used to immu-niz Balb/c mice.By cell culture and monoclonal antibody of dilution method,mAb named 2F3,5C2 and 5C7 were prepared against avian leukosis virus p27 protein by fusing myeloma cell line SP2/0 with spleen lymphocytes of immunized Balb/c mice.By biological characteristic analysis,the number of three strain hybridoma cell chromosomes was about 90.Then immunological identifications were conducted by means of ELISA/Western blot and IFA,the three mAb against p27 protein of ALV possessed high antibody ti-ters and good specificity.The result showed that the three McAbs had good reactogenicity.These three McAbs can be used as a valuable tool for avian leukosis virus epitope analysis,the detection of the virus and Kit research.%以纯化的禽白血病病毒 p27重组蛋白作为免疫原,按常规方法免疫 Balb/c 小鼠,取其脾细胞与 SP2/0细胞融合,通过细胞培养和有限稀释法制备单克隆抗体,最终获得了3株能稳定分泌禽白血病病毒 p27蛋白单克隆抗体的细胞株2F3、5C2和5C7。经生物学特性分析,3株杂交瘤细胞染色体数均为90条左右,通过 ELISA、Western blot 和间接免疫荧光等方法对所获得的3株单克隆抗体进行了鉴定,证明3株单克隆抗体细胞株所分泌的抗体效价高,而且均能够和禽白血病病毒 p27蛋白特异性发生反应。说明该3株单克隆抗体均具有与禽白血病病毒 p27蛋白发生反应的反应原性。3株单克隆抗体的成功获得为禽白血病病毒抗原表位的分析,病毒的检测以及试剂盒的研发奠定了基础。

  10. Uncoupling clutch size, prolactin, and luteinizing hormone using experimental egg removal.

    Science.gov (United States)

    Ryan, Calen P; Dawson, Alistair; Sharp, Peter J; Williams, Tony D

    2015-03-01

    Clutch size is a key avian fitness and life history trait. A physiological model for clutch size determination (CSD), involving an anti-gonadal effect of prolactin (PRL) via suppression of luteinizing hormone (LH), was proposed over 20 years ago, but has received scant experimental attention since. The few studies looking at a PRL-based mechanistic hypothesis for CSD have been equivocal, but recent experiments utilizing a pharmacological agent to manipulate PRL in the zebra finch (Taeniopygia guttata) found no support for a role of this hormone in clutch size determination. Here, we take a complementary approach by manipulating clutch size through egg removal, examining co-variation in PRL and LH between two breeding attempts, as well as through experimentally-extended laying. Clutch size increased for egg removal females, but not controls, but this was not correlated with changes in PRL or LH. There were also no differences in PRL between egg removal females and controls, nor did PRL levels during early, mid- or late-laying of supra-normal clutches predict clutch size. By uncoupling PRL, LH and clutch size in our study, several key predictions of the PRL-based mechanistic model for CSD were not supported. However, a positive correlation between PRL levels late in laying and days relative to the last egg (clutch completion) provides an alternative explanation for the equivocal results surrounding the conventional PRL-based physiological model for CSD. We suggest that females coordinate PRL-mediated incubation onset with clutch completion to minimize hatching asynchrony and sibling hierarchy, a behavior that is amplified in females laying larger clutches.

  11. In vitro expression of native H5 and N1 genes of avian influenza virus by using Green Fluorescent Protein as reporter

    Directory of Open Access Journals (Sweden)

    Risza Hartawan

    2011-10-01

    Full Text Available The hemagglutinin and neuraminidase are important immunogen of avian influenza virus that are suitable for recombinant experimentation. However, both genes have been experienced rapid mutation resulting in diverse variety of genotypes. Hence, gene expression in recombinant systems will be difficult to predict. The objective of the study was to examine expression level of H5 and N1 genes from a field isolate by cloning the genes into expression vector pEGFP-C1. Two clones respresenting fulllength of H5 and N1 gene in plasmid pEGFP-C1 were transfected into chicken embryo fibroblasts (CEF, rabbit kidney (RK13 and African green monkey kidney (VERO cells using Lipofectamine ‘Plus’ reagent. The experiment showed level of gene expression in the VERO cell was higher than in the RK13 and CEF cells. Observations using fluorescent microscopy and Western blotting revealed that the N1 gene was expressed better in all cells compared to the H5 gene.

  12. 罗格列酮对2型糖尿病并发非酒精性脂肪肝大鼠肝脏解偶联蛋白-2表达的影响%Effect of rosiglitazone on the expression of hepatic uncoupling protein 2 in type 2 diabetic rats complicated with non-alcoholic fatty liver disease

    Institute of Scientific and Technical Information of China (English)

    陈玉华; 武革; 郭中秋; 谭娅琴; 李善清; 陈晓铭

    2011-01-01

    Objective To observe the effect of rosiglitazone on the expressions of uncoupling protein-2 ( UCP-2 ) in the liver.of type 2 diabetic rats complicated with non-alcoholic fatty liver disease (NAFLD), and explore its mechanism of treating type 2 diabetes mellitus (T2DM) complicated with NAFLD.Methods The model rats of T2DM complicated with NAFLD were established by feeding high-glucose and high-fat diet, and injection of low dose streptozotocin.The model rats were randomly divided into 2 groups: model group (n = 16) and rosiglitazone group (n = 16).Twenty normal rats were set for normal control group.Intragastric administration lasted for 12 weeks.At the end of 16 weeks (after treated 8 weeks) and 20 weeks (after treated 12 weeks), the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), fasting blood glucose (FBG), fasting insulin (FINs), triglyceride (TG), total cholesterol (TC) and free fatty acid (FFA) in each group were tested and the level of insulin sensitivity index (ISI) was calculated.Meanwhile, pathological changes of liver, the expression of UCP-2 mRNA and UCP-2 protein of liver tissues in each group were detected.Results The levels of serum ALT, AST, FBG, FINs, serum TG, TC and FFA were significantly higher ( all P < 0.01 ), ISI was signifcantly lower ( P < 0.01 ), liver cirrhosis was significantly exacerbated, and the expressions of UCP-2 mRNA and UCP-2 protein were signifcantly increased (all P <0.01 ) in model group compared with control group.After treatment for 8 weeks and 12 weeks with rosiglitazone, the levels of serum ALT, AST, FBG, serum TG, TC and FFA were signifcantly decreaseed, ISI was signifcantly increased (P < 0.01 or P < 0.05 ), liver cirrhosis was significantly improved, and the expressions of UCP-2 mRNA and UCP-2 protein were signifcantly decreased (P <0.01 ) compared with model rats.Conclusion Rosiglitazone can control the appropriate expression of liver tissue UCP2 mRNA and UCP-2 protein of

  13. 解偶联蛋白-2、PPARγ在梗阻性黄疸及胆道再通大鼠肝脏氧化损伤中的作用%Roles of uncoupling protein-2 and peroxisome proliferators activated receptors γ in liver oxidative injuy of rats with obstructive jaundice and recannalization of biliary tract

    Institute of Scientific and Technical Information of China (English)

    李蔚; 邬善敏; 温暖; 陈辰; 蔡杰

    2013-01-01

    目的 研究解偶联蛋白-2(UCP-2)、过氧化物酶体增殖物激活受体γ(PPARγ)在梗阻性黄疸及胆道再通大鼠肝脏中的表达与肝功能及超微结构改变的关系.方法 将30只雄性Wistar大鼠随机分为3组:假手术组(A组)、梗阻性黄疸1周组(B组)、梗阻性黄疸1周胆道再通1周组(C组),检测各组大鼠血清中直接胆红素(DBIL)、总胆红素(TBIL)和丙氨酸转氨酶(ALT)水平.免疫组织化学法检测UCP-2、PPARγ蛋白在肝脏中的表达.结果 血清DBIL、TBIL、ALT水平:B组明显高于A组,C组明显低于B组.PPARγ的表达:B组明显低于A组,C组较B组明显增强;UCP-2的表达:B组明显强于A组,C组较B组明显减弱.在电镜下可见B组超微结构改变较A组显著,C组超微结构改变较B组明显减轻.结论 梗阻性黄疸氧化损伤,造成肝功能及超微结构的改变,解除梗阻有利于肝功能及超微结构的恢复.PPARγ可能参与梗阻性黄疸及胆道再通肝脏的氧化与抗氧化反应,UCP-2可能参与肝脏氧化损伤时氧自由基的清除,减轻肝脏氧化损伤.%Objective To investigate the relationship between the expression of uncoupling protein-2 (UCP-2) and proliferators activated receptors y (PPARy) with the liver function and ultrastruc-tural changes in rats with obstructive jaundice and bile flow restoration rates, and to explore the mechanisms of oxidative injuy in hepatic obstructive jaundice. Methods Thirty healthy male Wistar rats were divided into three groups at random: group A (sham group), group B (obstructive jaundice one-week group), group C (obstructive jaundice one-week and recanalization one-week group). The levels of direct bilirubin (DBIL), total bilirubin (TBIL) and serum alanine aminotransferase (ALT) were measured. The expression levels of UCP-2 and PPARy proteins in hepatic tissues were detected by u-sing immunohistochemistry. Results The levels of serum DBIL, TBIL and ALT in group B were increased significantly as

  14. 稳定表达新城疫病毒V蛋白禽源细胞系的建立%Establishment of stable avian cell line expressing V protein of Newcastle disease virus

    Institute of Scientific and Technical Information of China (English)

    唐文雅; 王兴龙; 张渭东; 杜恩岐; 陈胜利; 王丹阳; 党如意; 张淑霞; 杨增岐

    2012-01-01

    为获得可以稳定表达新城疫病毒V蛋白的DF-1细胞系,以质粒载体为模板,用PCR方法扩增新城疫病毒V基因并引入酶切位点(EcoRⅠ和BamHⅠ),以同尾酶相连的方式与用EcoRⅠ和BglⅡ酶切的piggyBac转座子系统克隆质粒pTKL1-CMV-Puro-EGFP相连接,经酶切鉴定、测序分析正确后,用Lipo-fectamineTM2000将重组质粒pTKL1-CMV-Puro-EGFP-V和辅助质粒mPB共转染DF-1细胞,通过绿色荧光观察、嘌呤霉素筛选及蛋白表达的Western-blot检测,结果表明,稳定表达新城疫病毒V蛋白的DF-1细胞系已成功建立。证实piggyBac转座子系统能够用于稳定表达外源基因的禽源细胞系的构建,为piggy-Bac转座子系统在禽源细胞的应用和新城疫病毒V蛋白的研究提供了一定的参考资料。%In order to obtain the stable DF-1 cell lines expressing Newcastle disease virus(NDV) V protein,NDV V gene from the recombinant plasmid pEGFT-N1-V was amplified, the restriction sites of EcoR l and BamH I were inducted into the amplicon. The amplicon were then connected into the piggy- Bac transposon cloning plasmid pTKL1-CMV-Puro-EGFP vector,which was checked by enzyme digestion and sequence analysis. The recombinant plasmids were co-transfected with the helper plasmid mPB into DF-1 cells using LipofectamineTM2000. The cell line was selected with puromycin and detected with green fluorescent protein observation, and V protein expression was detected by Western-blot. Results showed that the stable monoclonal cell lines expressing V protein was constructed successfully. The results revealed that piggyBac transposon system can be used for the construction of avian stable cell lines and the DF-1 cell line(DF-1-V) expressing NDV V protein was established successfully. The present study provided reference materials for application of the piggyBac transposon system in avian cell and research of NDV V protein.

  15. 甲状腺激素减少对大鼠肾脏解耦联蛋白-2表达的影响%Study on the expression of uncoupling protein 2 in the kidney of the rats with hypothyroidism

    Institute of Scientific and Technical Information of China (English)

    许静; 田秀标; 房辉; 刘鹏; 孟庆义

    2010-01-01

    目的 观察甲状腺激素减少时肾脏线粒体解耦联蛋白-2(UCP2)表达的改变,探讨甲状腺激素减少时肾脏损伤的发生机制. 方法 按随机数字表法将Wistar大鼠分为对照组和甲状腺激素减少组(甲减组),每组10只.采用低碘饮食复制甲状腺功能减退大鼠模型,对照组摄碘量10.00 μg/d,甲减组1.24 μg/d,两组均适应喂养1周,条件饲养3个月后取血测定甲状腺功能指标,取肾脏组织,用免疫组化法及逆转录-聚合酶链反应(RT-PCR)测定UCP2的蛋白及mRNA表达. 结果 与对照组比较,甲减组促甲状腺激素(TSH,mU/L)明显上升(4.88±1.37比1.65±0.33,P<0.05),三碘甲状腺原氨酸总量(TT3)、甲状腺素总量(TT4)、游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)均明显下降[TT3(nmol/L):0.54±0.07比0.98±0.17;TT4(nmol/L):7.82±2.18比48.78±3.65;FT3(pmol/L):2.28±0.22比2.99±0.10;FT4(pmol/L):11.38±1.74比29.27±0.95,均P<0.01].免疫组化显示,甲减组UCP2蛋白在肾小球和肾小管中的表达均低于对照组(肾小球:0.17±0.02比0.24±0.04,肾小管:0.19±0.02比0.25±0.02,均P<0.01).RT-PCR显示,甲减组UCP2 mRNA表达明显低于对照组(0.70±0.19比1.30±0.09,P<0.01). 结论 甲状腺激素减少时肾脏中UCP2表达下调,提示可能会有损伤肾脏,其机制与肾脏线粒体UCP2表达下调有关.%Objective To observe the change in the expression of uncoupling protein 2(UCP2) in kidney mitochondria in rats with hypothyroidism,and study the mechanism of renal injury due to hypothyroidism. Methods The Wistar rats were randomly divided into control group(n=10) and hypothyroidism group(n=10).The hypothyroidism rat model was reproduced by low-iodine diet.The intake of iodine in control group and hypothyroidism group were 10.00 μg/d and 1.24 μg/d,respectively.The rats were raised under these conditions respectively for 3 months after they adapted to the feeding for 1 week.Then the thyroid function parameters were measured

  16. Study on the Beam Quality of Uncoupled Laser Diode Arrays

    Institute of Scientific and Technical Information of China (English)

    GAO Chunqing; WEI Guanghui

    2001-01-01

    The beam quality of uncoupled laser diode array is studied theoretically and experimentally. By calculating the second order moments of the beam emitted from the laser diode array, the dependence of the M2-factor of the laser diode array on the M2-factor of the single emitter, the ratio of the emitting region to the non-emitting space, and the number of emitters, has been deduced. From the measurement of the beam propagation the M2-factor of a laser diode bar is experimentally determined. The measured M2-factor of the laser diode bar agrees with the theoretical prediction.

  17. Relationship between the expression of uncoupling protein 2 and the damage by oxygen free radicals in acute liver failure rats%急性肝衰竭大鼠肝脏解偶联蛋白2的表达及其与氧化损伤的关系

    Institute of Scientific and Technical Information of China (English)

    刘均艳; 赵海红; 朱敏; 陈景丹; 朱坚胜

    2011-01-01

    Objective To investigate the relationship between uncoupling protein 2 (UCP2) expression and the damage caused by oxygen free radicals in acute liver failure rat models. Methods Thirty-five male Sprague-Dawley rats were randomly divided into two groups: the control group (15 rats) and liver failure group (20 rats). The rats were injected intraperitoneally with thioacetamide (TAA) to induce models of acute liver failure. The levels of endotoxin (ET) were detected by double antibody sandwich enzyme-linked imrnunosorbent assay. The expression of liver UCP2 mRNA was detected by reverse transcription polymerase chain reactin. The superoxide dismutase (SOD) and malonaldehyde (MDA) were detected by spectrophotometry. The expression of UCP2 protein was observed by immunohistochemistry. The data of the two groups were compared using Mann-Whitney U test or ANOVA. Results The expression of UCP2 mRNA in liver failure group was higher as compared to the control group (P < 0.01); the level of MDA and endotoxin of liver failure group were higher than that of the control group (P < 0.01). SOD of the liver failure group was lower (P < 0.01). There was a certain correlation between UCP2 mRNA expression and ET, SOD and MDA (r = 0.952, -0.667, 0. 634 respectively, P < 0.05 or 0.01). Conclusions UCP2 is highly expressed in the livers of liver failure rats. A certain correlation perhaps existed between the expression of UCP2 mRNA and the seral SOD, MDA and ET.%目的 研究急性肝衰竭大鼠肝脏解偶联蛋白(UCP)2的表达及其与内毒索、氧自由基损伤的关系,进一步探讨肝衰竭的发病机制.方法将SD大鼠35只随机分为对照组(15只)、肝衰竭组(20只).腹腔注射硫代乙酰胺(TAA)制备肝衰竭大鼠模型.测定各组大鼠血清内毒素(ET)水平,肝脏组织UCP2 mRNA的表达、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性,免疫组织化学观察UCP2表达.组间比较采用Mann-Whitney U检验或单因素

  18. Carboxyatractyloside effects on brown-fat mitochondria imply that the adenine nucleotide translocator isoforms ANT1 and ANT2 may be responsible for basal and fatty-acid-induced uncoupling respectively.

    Science.gov (United States)

    Shabalina, Irina G; Kramarova, Tatiana V; Nedergaard, Jan; Cannon, Barbara

    2006-11-01

    In brown-fat mitochondria, fatty acids induce thermogenic uncoupling through activation of UCP1 (uncoupling protein 1). However, even in brown-fat mitochondria from UCP1-/- mice, fatty-acid-induced uncoupling exists. In the present investigation, we used the inhibitor CAtr (carboxyatractyloside) to examine the involvement of the ANT (adenine nucleotide translocator) in the mediation of this UCP1-independent fatty-acid-induced uncoupling in brown-fat mitochondria. We found that the contribution of ANT to fatty-acid-induced uncoupling in UCP1-/- brown-fat mitochondria was minimal (whereas it was responsible for nearly half the fatty-acid-induced uncoupling in liver mitochondria). As compared with liver mitochondria, brown-fat mitochondria exhibit a relatively high (UCP1-independent) basal respiration ('proton leak'). Unexpectedly, a large fraction of this high basal respiration was sensitive to CAtr, whereas in liver mitochondria, basal respiration was CAtr-insensitive. Total ANT protein levels were similar in brown-fat mitochondria from wild-type mice and in liver mitochondria, but the level was increased in brown-fat mitochondria from UCP1-/- mice. However, in liver, only Ant2 mRNA was found, whereas in brown adipose tissue, Ant1 and Ant2 mRNA levels were equal. The data are therefore compatible with a tentative model in which the ANT2 isoform mediates fatty-acid-induced uncoupling, whereas the ANT1 isoform may mediate a significant part of the high basal proton leak in brown-fat mitochondria.

  19. Vaccines and vaccination for avian influenza in poultry

    Science.gov (United States)

    Avian influenza (AI) vaccines have been developed and used to protect poultry and other birds in various countries of the world. Protection is principally mediated by an immune response to the subtype-specific hemagglutinin (HA) protein. AI vaccines prevent clinical signs of disease, death, egg pr...

  20. Comparative genomic data of the Avian Phylogenomics Project

    DEFF Research Database (Denmark)

    Zhang, Guojie; Li, Bo; Li, Cai;

    2014-01-01

    , which include 38 newly sequenced avian genomes plus previously released or simultaneously released genomes of Chicken, Zebra finch, Turkey, Pigeon, Peregrine falcon, Duck, Budgerigar, Adelie penguin, Emperor penguin and the Medium Ground Finch. We hope that this resource will serve future efforts...... in an average N50 scaffold size of about 50 kb. Repetitive elements comprised 4%-22% of the bird genomes. The assembled scaffolds allowed the homology-based annotation of 13,000 ~ 17000 protein coding genes in each avian genome relative to chicken, zebra finch and human, as well as comparative and sequence...

  1. The Helper Activities of Different Avian Viruses for Propagation of Recombinant Avian Adeno-Associated Virus

    Institute of Scientific and Technical Information of China (English)

    WANG An-ping; SUN Huai-chang; WANG Jian-ye; WANG Yong-juan; YUAN Wei-feng

    2007-01-01

    To compare the helper activities of different avian viruses for propagation of recombinant avian adeno-associated virus (rAAAV), AAV-293 cells were cotransfected with the AAAV vector pAITR-GFP containing green fluorescent protein (GFP) gene, the AAAV helper vector pcDNA-ARC expressing the rep and cap genes, and the adenovirus helper vector pHelper expressing Ad5 E2A, E4, and VA-RNA genes. Chicken embryonic fibroblast (CEF) or chicken embryonic liver (CEL) cells were cotransfected with the AAAV vector and the AAAV helper vector, followed by infection with Marek's disease virus (MDV), avian adenovirus, chicken embryo lethal orphan (CELO) virus or infectious bursal disease virus (IBDV). Infectious rAAAV particles generated by the two strategies were harvested and titrated on CEF and CEL cells. A significantly higher viral titer was obtained with the helper activity provided by the pHelper vector than by MDV or CELO virus. Further experiments showed that rAAAV-mediated green fluorescent protein (gfp) expression was overtly enhanced by MDV or CELO virus super infection or treatment with sodium butyric acid, but not by IBDV super infection. These data demonstrated that MDV and CELO viruses could provide weak helper activity for propagation of rAAAV, and rAAAV-mediated transgene expression could be enhanced by super infection with the helper viruses.

  2. Chimera states in uncoupled neurons induced by a multilayer structure

    CERN Document Server

    Majhi, Soumen; Ghosh, Dibakar

    2016-01-01

    Spatial coexistence of coherent and incoherent dynamics in network of coupled oscillators is called a chimera state. We study such chimera states in a network of neurons without any direct interactions but connected through another medium of neurons, forming a multilayer structure. The upper layer is thus made up of uncoupled neurons and the lower layer plays the role of a medium through which the neurons in the upper layer share information among each other. Hindmarsh-Rose neurons with square wave bursting dynamics are considered as nodes in both layers. In addition, we also discuss the existence of chimera states in presence of inter layer heterogeneity. The neurons in the bottom layer are globally connected through electrical synapses, while across the two layers chemical synapses are formed. According to our research, the competing effects of these two types of synapses can lead to chimera states in the upper layer of uncoupled neurons. Remarkably, we find a density-dependent threshold for the emergence o...

  3. H9N2 Avian Influenza Virus Protein PB1 Enhances the Immune Responses of Bone Marrow-Derived Dendritic Cells by Down-Regulating miR375

    Science.gov (United States)

    Lin, Jian; Xia, Jing; Tu, Chong Z.; Zhang, Ke Y.; Zeng, Yan; Yang, Qian

    2017-01-01

    Polymerase basic protein 1 (PB1), the catalytic core of the influenza A virus RNA polymerase complex, is essential for viral transcription and replication. Dendritic cells (DCs) possess important antigen presenting ability and a crucial role in recognizing and clearing virus. MicroRNA (miRNA) influence the development of DCs and their ability to present antigens as well as the ability of avian influenza virus (AIV) to infect host cells and replicate. Here, we studied the molecular mechanism underlying the miRNA-mediated regulation of immune function in mouse DCs. We first screened for and verified the induction of miRNAs in DCs after PB1 transfection. Results showed that the viral protein PB1 down-regulated the expression of miR375, miR146, miR339, and miR679 in DCs, consistent with the results of H9N2 virus treatment; however, the expression of miR222 and miR499, also reduced in the presence of PB1, was in contrast to the results of H9N2 virus treatment. Our results suggest that PB1 enhanced the ability of DCs to present antigens, activate lymphocytes, and secrete cytokines, while miR375 over-expression repressed activation of DC maturation. Nevertheless, PB1 could not promote DC maturation once miR375 was inhibited. Finally, we revealed that PB1 inhibited the P-Jnk/Jnk signaling pathway, but activated the p-Erk/Erk signaling pathway. While inhibition of miR375 -activated the p-Erk/Erk and p-p38/p38 signaling pathway, but repressed the P-Jnk/Jnk signaling pathway. Taken together, results of our studies shed new light on the roles and mechanisms of PB1 and miR375 in regulating DC function and suggest new strategies for combating AIV.

  4. Avian Reovirus Protein p17 Functions as a Nucleoporin Tpr Suppressor Leading to Activation of p53, p21 and PTEN and Inactivation of PI3K/AKT/mTOR and ERK Signaling Pathways.

    Directory of Open Access Journals (Sweden)

    Wei-Ru Huang

    Full Text Available Avian reovirus (ARV protein p17 has been shown to regulate cell cycle and autophagy by activation of p53/PTEN pathway; nevertheless, it is still unclear how p53 and PTEN are activated by p17. Here, we report for the first time that p17 functions as a nucleoporin Tpr suppressor that leads to p53 nuclear accumulation and consequently activates p53, p21, and PTEN. The nuclear localization signal (119IAAKRGRQLD128 of p17 has been identified for Tpr binding. This study has shown that Tpr suppression occurs by p17 interacting with Tpr and by reducing the transcription level of Tpr, which together inhibit Tpr function. In addition to upregulation of PTEN by activation of p53 pathway, this study also suggests that ARV protein p17 acts as a positive regulator of PTEN. ARV p17 stabilizes PTEN by stimulating phosphorylation of cytoplasmic PTEN and by elevating Rak-PTEN association to prevent it from E3 ligase NEDD4-1 targeting. To activate PTEN, p17 is able to promote β-arrestin-mediated PTEN translocation from the cytoplasm to the plasma membrane via a Rock-1-dependent manner. The accumulation of p53 in the nucleus induces the PTEN- and p21-mediated downregulation of cyclin D1 and CDK4. Furthermore, Tpr and CDK4 knockdown increased virus production in contrast to depletion of p53, PTEN, and LC3 reducing virus yield. Taken together, our data suggest that p17-mediated Tpr suppression positively regulates p53, PTEN, and p21 and negatively regulates PI3K/AKT/mTOR and ERK signaling pathways, both of which are beneficial for virus replication.

  5. Avian Reovirus Protein p17 Functions as a Nucleoporin Tpr Suppressor Leading to Activation of p53, p21 and PTEN and Inactivation of PI3K/AKT/mTOR and ERK Signaling Pathways.

    Science.gov (United States)

    Huang, Wei-Ru; Chiu, Hung-Chuan; Liao, Tsai-Ling; Chuang, Kuo-Pin; Shih, Wing-Ling; Liu, Hung-Jen

    2015-01-01

    Avian reovirus (ARV) protein p17 has been shown to regulate cell cycle and autophagy by activation of p53/PTEN pathway; nevertheless, it is still unclear how p53 and PTEN are activated by p17. Here, we report for the first time that p17 functions as a nucleoporin Tpr suppressor that leads to p53 nuclear accumulation and consequently activates p53, p21, and PTEN. The nuclear localization signal (119IAAKRGRQLD128) of p17 has been identified for Tpr binding. This study has shown that Tpr suppression occurs by p17 interacting with Tpr and by reducing the transcription level of Tpr, which together inhibit Tpr function. In addition to upregulation of PTEN by activation of p53 pathway, this study also suggests that ARV protein p17 acts as a positive regulator of PTEN. ARV p17 stabilizes PTEN by stimulating phosphorylation of cytoplasmic PTEN and by elevating Rak-PTEN association to prevent it from E3 ligase NEDD4-1 targeting. To activate PTEN, p17 is able to promote β-arrestin-mediated PTEN translocation from the cytoplasm to the plasma membrane via a Rock-1-dependent manner. The accumulation of p53 in the nucleus induces the PTEN- and p21-mediated downregulation of cyclin D1 and CDK4. Furthermore, Tpr and CDK4 knockdown increased virus production in contrast to depletion of p53, PTEN, and LC3 reducing virus yield. Taken together, our data suggest that p17-mediated Tpr suppression positively regulates p53, PTEN, and p21 and negatively regulates PI3K/AKT/mTOR and ERK signaling pathways, both of which are beneficial for virus replication.

  6. Influence of uncoupling protein 2 gene expression to the dysfunction of beta cells caused by increased free fatty acid%解偶联蛋白2基因表达对游离脂肪酸升高所致β细胞功能障碍的影响

    Institute of Scientific and Technical Information of China (English)

    王冰; 李宏亮; 杨文英; 肖建中; 杜瑞琴; 白秀平

    2013-01-01

    infusion in unrestrained rats was used for fat emulsion and heparin or saline infusion.The infusion period started on day 2 after surgery.After 48 h infusion,fasting serum insulin(INS),venous free fat acid were determined.The intravenous glucose tolerance test and islet cell perifusion were conducted to evaluate the function of islet β cell.The rats in the two groups were sacrificed,and the pancreatic islets were isolated and collected.The expression of insulin receptor substrate-1 (IRS-1),insulin receptor substrate-2 (IRS-2),uncoupling protein-2 (UCP2) gene in islets were detected by real-time polymerase chain reaction (PCR).Student's t test was used for data analysis.Results The serum FFA and insulin concentration of blood in FFA group were higher than those in NS group (insulin (25.2 ± 2.3) vs (18.6±1.7)mU/L,t=7.9,P<0.05,FFA (1.39 ±0.18) vs (0.64 ± 0.10)mmol/L,t=12.8,P <0.05).The glucose stimulated insulin secretion increased in the FFA group than in NS group ((137 ±24) vs (80 ± 16) mU/L in vivo,t =6.8,P < 0.05 ; (272 ± 4) vs (227 ± 4) mU/L in vitro,t =28.6,P <0.05).The gene expression of IRS-1 in islets was significantly increased by 29.3% ± 2.6% (t =2.2,P <0.05),and the mRNA expression of IRS-2,UCP-2 increased by 345.1% ±4.7% and 228.4% ±4.2% in FFA group than those in NS group (t =3.4,3.0,all P < 0.05).Conclusion Lipid infusion in short-term increases the secretion of insulin but it can also increase the gene expression of UCP-2,which can damage islet β cells.

  7. Dynamic expression of uncoupling protein 2 in rats models of acute liver failure and its significance%解耦联蛋白2在急性肝功能衰竭大鼠肝脏中的动态表达和意义

    Institute of Scientific and Technical Information of China (English)

    肖二辉; 陈永平; 戴志娟; 张磊; 张晓华; 谷甸娜

    2009-01-01

    Objective To explore the expression and significance of uncoupling protein (UCP)2in rats models of acute liver failure (ALF). Methods Thirty-six healthy male SD rats were randomly divided into normal control group and model group, and the model group was divided into 5 subgroups:6, 12, 24, 36 and 48 hours sub groups with 6 rats in each sub group. The rat model of ALF was established by intraperitoneal injections of D-galactosamine (D-Gal) and lipopolysaccharide (LPS).Sections of liver tissue were stained with hematoxylin and eosin and observed under optical microscope.UCP2 and UCP2 mRNA in rat liver were determined at different time points with immunohistochemical method and reverse transcription-polymerase chain reaction ( RT-PCR ),respectively. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and malondialdehyde (MDA) concentration in the liver tissues were analyzed at the same time points.Comparisons among all the experimental groups were done by SNK test. Results Infiltration of inflammatory cells and necrosis of hepatic cells were marked in model group,and ALT, AST and MDA in model group were significantly higher than those in control group [(24. 0 ± 2. 0) U/L, (82. 3±16. 9) U/L, (2. 55±0. 22)μmol/g] at all time points. And they reached a peak at 24 h [(8346. 7±1363. 1) U/L, (9766. 7±1274. 1) U/L, (8. 34±1. 13) μmol/g; all P<0. 05]. UCP2 and UCP2 mRNA expressed scarcely in the liver tissues of control group, while increased markedly from 6 to 48 hours after D-Gal/LPS challenge in model group (P<0. 05). They both reached a peak at 24 h. And the discrepancy between consecutive experimental group had statistical significance ( P < 0. 05).Conclusions The rat model of ALF was established successfully by intraperitoneal injections of D-gal and LPS. The expression levels of UCP2 mRNA and UCP2 are consistent with the extent of liver injury and the level of oxidative stress in the rat model of ALF.%目的 探讨急性肝功能

  8. Expression of uncoupling protein-2 and induced nitric oxide synthase in enalapril preprocessing rat myocardium%依那普利预处理对大鼠心肌组织解偶联蛋白2和诱导型一氧化氮合酶表达的影响

    Institute of Scientific and Technical Information of China (English)

    康丹丹; 汤建民

    2014-01-01

    目的:研究依那普利预处理对大鼠心肌缺血再灌注损伤时心肌组织解偶联蛋白2(UCP2)和诱导型一氧化氮合酶( iNOS)表达的影响,并探讨其可能的作用机制。方法:将35只SD大鼠随机分成3组,假手术组7只、缺血再灌注组14只、依那普利预处理组14只。假手术组只穿线不结扎,缺血再灌注组结扎30 min后,再灌注120 min;依那普利预处理组结扎前用依那普利10 mg/( kg· d)灌胃1周,余处理方法同缺血再灌注组。实验终末采血测血清磷酸激酶(CK)活性及丙二醛(MDA)含量;取3组大鼠左室缺血区心肌组织,采用 RT-PCR和Western blot检测左室心肌组织UCP2和iNOS的表达。结果:3组大鼠间血清CK活性、MDA含量及心肌组织UCP2、iNOS表达量比较,差异均有统计学意义(F=1523.311、32.124、203.504、235.933、65.794、113.630,P均<0.001)。缺血再灌注组与假手术组相比UCP2、iNOS表达升高,血清CK活性、MDA含量升高(P均<0.05);预处理组与缺血再灌注组相比UCP2表达下降,iNOS表达升高,血清CK活性、MDA含量降低( P均<0.05)。结论:依那普利预处理对大鼠心肌缺血再灌注损伤的保护作用可能与其抑制UCP2表达、增强iNOS表达有关。%Aim:To investigate the changes of uncoupling protein-2(UCP2)and induced nitric oxide synthase (iNOS) expression following myocardial ischemia reperfusion ( IR ) injury in rats treated with enalapril , and observe the possible mechanism.Methods:Thirty-five SD rats were randomly assigned into three groups:Sham group(n=7),IR group(n=14),and PIP group(n=14).In the IR group,the rats were subjected to 30 minutes of LAD ligation and subsequent reperfu-sion for 120 min.In the Sham group ,the rats underwent the similar surgical procedure without ligation of LAD .In the PIP group,the rats received one week of pretreatment with enalapril ,and then underwent IR

  9. Non structural protein of avian influenza A (H11N1 virus is a weaker suppressor of immune responses but capable of inducing apoptosis in host cells

    Directory of Open Access Journals (Sweden)

    Mukherjee Sanjay

    2012-08-01

    Full Text Available Abstract Background The Non-Structural (NS1 protein of Influenza A viruses is an extensively studied multifunctional protein which is commonly considered as key viral component to fight against host immune responses. Even though there has been a lot of studies on the involvement of NS1 protein in host immune responses there are still ambiguities regarding its role in apoptosis in infected cells. Interactions of NS1 protein with host factors, role of NS1 protein in regulating cellular responses and apoptosis are quite complicated and further studies are still needed to understand it completely. Results NS1 genes of influenza A/Chicken/India/WBNIV2653/2008 (H5N1 and A/Aquatic bird/India/NIV-17095/2007(H11N1 were cloned and expressed in human embryonic kidney (293T cells. Microarray based approach to study the host cellular responses to NS1 protein of the two influenza A viruses of different pathogenicity showed significant differences in the host gene expression profile. NS1 protein of H5N1 resulted in suppression of IFN-β mediated innate immune responses, leading to down-regulation of the components of JAK-STAT pathway like STAT1 which further suppressed the expression of pro-inflammatory cytokines like CXCL10 and CCL5. The degree of suppression of host immune genes was found considerable with NS1 protein of H11N1 but was not as prominent as with H5N1-NS1. TUNEL assay analyses were found to be positive in both the NS1 transfected cells indicating both H5N1 as well as H11N1 NS1 proteins were able to induce apoptosis in transfected cells. Conclusions We propose that NS1 protein of both H5N1 and H11N1 subtypes of influenza viruses are capable of influencing host immune responses and possess necessary functionality to support apoptosis in host cells. H11N1, a low pathogenic virus without any proven evidence to infect mammals, contains a highly potential NS1 gene which might contribute to greater virus virulence in different gene combinations.

  10. Koyukuk NWR 1985 avian checklist

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — An avian checklist survey was conducted within the boundaries of the Koyukuk National Wildlife Refuge and Kaiyuh Flats unit of the Innoko National Wildlife Refuge...

  11. Koyukuk NWR 1986 avian checklist

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — An avian checklist survey was conducted within the boundaries of the Koyukuk National Wildlife Refuge and Kaiyuh Flats unit of the Innoko National Wildlife Refuge in...

  12. Simulating avian wingbeat kinematics.

    Science.gov (United States)

    Parslew, Ben; Crowther, William J

    2010-12-01

    Inverse dynamics methods are used to simulate avian wingbeats in varying flight conditions. A geometrically scalable multi-segment bird model is constructed, and optimisation techniques are employed to determine segment motions that generate desired aerodynamic force coefficients with minimal mechanical power output. The results show that wingbeat kinematics vary gradually with changes in cruise speed, which is consistent with experimental data. Optimised solutions for cruising flight of the pigeon suggest that upstroke wing retraction is used as a method of saving energy. Analysis of the aerodynamic force coefficient variation in high and low speed cruise leads to the proposal that a suitable gait metric should include both thrust and lift generation during each half-stroke.

  13. Avian host defense peptides.

    Science.gov (United States)

    Cuperus, Tryntsje; Coorens, Maarten; van Dijk, Albert; Haagsman, Henk P

    2013-11-01

    Host defense peptides (HDPs) are important effector molecules of the innate immune system of vertebrates. These antimicrobial peptides are also present in invertebrates, plants and fungi. HDPs display broad-spectrum antimicrobial activities and fulfill an important role in the first line of defense of many organisms. It is becoming increasingly clear that in the animal kingdom the functions of HDPs are not confined to direct antimicrobial actions. Research in mammals has indicated that HDPs have many immunomodulatory functions and are also involved in other physiological processes ranging from development to wound healing. During the past five years our knowledge about avian HDPs has increased considerably. This review addresses our current knowledge on the evolution, regulation and biological functions of HDPs of birds.

  14. BIRD FLU (AVIAN INFLUENZA

    Directory of Open Access Journals (Sweden)

    Ali ACAR

    2005-12-01

    Full Text Available Avian influenza (bird flu is a contagious disease of animals caused by influenza A viruses. These flu viruses occur naturally among birds. Actually, humans are not infected by bird flu viruses.. However, during an outbreak of bird flu among poultry, there is a possible risk to people who have contact infect birds or surface that have been contaminated with excreations from infected birds. Symptoms of bird flu in humans have ranged from typical flu-like symptoms to eye infections, pneumonia, severe respiratory diseases and other severe and life-threatening complications. In such situation, people should avoid contact with infected birds or contaminated surface, and should be careful when handling and cooking poultry. [TAF Prev Med Bull 2005; 4(6.000: 345-353

  15. Isoproterenol Increases Uncoupling, Glycolysis, and Markers of Beiging in Mature 3T3-L1 Adipocytes.

    Science.gov (United States)

    Miller, Colette N; Yang, Jeong-Yeh; England, Emily; Yin, Amelia; Baile, Clifton A; Rayalam, Srujana

    2015-01-01

    Beta-adrenergic activation stimulates uncoupling protein 1 (UCP1), enhancing metabolic rate. In vitro, most work has studied brown adipocytes, however, few have investigated more established adipocyte lines such as the murine 3T3-L1 line. To assess the effect of beta-adrenergic activation, mature 3T3-L1s were treated for 6 or 48 hours with or without isoproterenol (10 and 100 μM) following standard differentiation supplemented with thyroid hormone (T3; 1 nM). The highest dose of isoproterenol increased lipid content following 48 hours of treatment. This concentration enhanced UCP1 mRNA and protein expression. The increase in UCP1 following 48 hours of isoproterenol increased oxygen consumption rate. Further, coupling efficiency of the electron transport chain was disturbed and an enhancement of glycolytic rate was measured alongside this, indicating an attempt to meet the energy demands of the cell. Lastly, markers of beige adipocytes (protein content of CD137 and gene transcript of CITED1) were also found to be upregulated at 48 hours of isoproterenol treatment. This data indicates that mature 3T3-L1 adipocytes are responsive to isoproterenol and induce UCP1 expression and activity. Further, this finding provides a model for further pharmaceutical and nutraceutical investigation of UCP1 in 3T3-L1s.

  16. Isoproterenol Increases Uncoupling, Glycolysis, and Markers of Beiging in Mature 3T3-L1 Adipocytes.

    Directory of Open Access Journals (Sweden)

    Colette N Miller

    Full Text Available Beta-adrenergic activation stimulates uncoupling protein 1 (UCP1, enhancing metabolic rate. In vitro, most work has studied brown adipocytes, however, few have investigated more established adipocyte lines such as the murine 3T3-L1 line. To assess the effect of beta-adrenergic activation, mature 3T3-L1s were treated for 6 or 48 hours with or without isoproterenol (10 and 100 μM following standard differentiation supplemented with thyroid hormone (T3; 1 nM. The highest dose of isoproterenol increased lipid content following 48 hours of treatment. This concentration enhanced UCP1 mRNA and protein expression. The increase in UCP1 following 48 hours of isoproterenol increased oxygen consumption rate. Further, coupling efficiency of the electron transport chain was disturbed and an enhancement of glycolytic rate was measured alongside this, indicating an attempt to meet the energy demands of the cell. Lastly, markers of beige adipocytes (protein content of CD137 and gene transcript of CITED1 were also found to be upregulated at 48 hours of isoproterenol treatment. This data indicates that mature 3T3-L1 adipocytes are responsive to isoproterenol and induce UCP1 expression and activity. Further, this finding provides a model for further pharmaceutical and nutraceutical investigation of UCP1 in 3T3-L1s.

  17. Uncoupling primer and releaser responses to pheromone in honey bees

    Science.gov (United States)

    Grozinger, Christina M.; Fischer, Patrick; Hampton, Jacob E.

    2007-05-01

    Pheromones produce dramatic behavioral and physiological responses in a wide variety of species. Releaser pheromones elicit rapid responses within seconds or minutes, while primer pheromones produce long-term changes which may take days to manifest. Honeybee queen mandibular pheromone (QMP) elicits multiple distinct behavioral and physiological responses in worker bees, as both a releaser and primer, and thus produces responses on vastly different time scales. In this study, we demonstrate that releaser and primer responses to QMP can be uncoupled. First, treatment with the juvenile hormone analog methoprene leaves a releaser response (attraction to QMP) intact, but modulates QMP’s primer effects on sucrose responsiveness. Secondly, two components of QMP (9-ODA and 9-HDA) do not elicit a releaser response (attraction) but are as effective as QMP at modulating a primer response, downregulation of foraging-related brain gene expression. These results suggest that different responses to a single pheromone may be produced via distinct pathways.

  18. Uncoupling of neurogenesis and differentiation during retinal development.

    Science.gov (United States)

    Engerer, Peter; Suzuki, Sachihiro C; Yoshimatsu, Takeshi; Chapouton, Prisca; Obeng, Nancy; Odermatt, Benjamin; Williams, Philip R; Misgeld, Thomas; Godinho, Leanne

    2017-03-03

    Conventionally, neuronal development is regarded to follow a stereotypic sequence of neurogenesis, migration, and differentiation. We demonstrate that this notion is not a general principle of neuronal development by documenting the timing of mitosis in relation to multiple differentiation events for bipolar cells (BCs) in the zebrafish retina using in vivo imaging. We found that BC progenitors undergo terminal neurogenic divisions while in markedly disparate stages of neuronal differentiation. Remarkably, the differentiation state of individual BC progenitors at mitosis is not arbitrary but matches the differentiation state of post-mitotic BCs in their surround. By experimentally shifting the relative timing of progenitor division and differentiation, we provide evidence that neurogenesis and differentiation can occur independently of each other. We propose that the uncoupling of neurogenesis and differentiation could provide neurogenic programs with flexibility, while allowing for synchronous neuronal development within a continuously expanding cell pool.

  19. Skeletal muscle mitochondrial uncoupling in a murine cancer cachexia model.

    Science.gov (United States)

    Tzika, A Aria; Fontes-Oliveira, Cibely Cristine; Shestov, Alexander A; Constantinou, Caterina; Psychogios, Nikolaos; Righi, Valeria; Mintzopoulos, Dionyssios; Busquets, Silvia; Lopez-Soriano, Francisco J; Milot, Sylvain; Lepine, Francois; Mindrinos, Michael N; Rahme, Laurence G; Argiles, Josep M

    2013-09-01

    Approximately half of all cancer patients present with cachexia, a condition in which disease-associated metabolic changes lead to a severe loss of skeletal muscle mass. Working toward an integrated and mechanistic view of cancer cachexia, we investigated the hypothesis that cancer promotes mitochondrial uncoupling in skeletal muscle. We subjected mice to in vivo phosphorous-31 nuclear magnetic resonance (31P NMR) spectroscopy and subjected murine skeletal muscle samples to gas chromatography/mass spectrometry (GC/MS). The mice used in both experiments were Lewis lung carcinoma models of cancer cachexia. A novel 'fragmented mass isotopomer' approach was used in our dynamic analysis of 13C mass isotopomer data. Our 31P NMR and GC/MS results indicated that the adenosine triphosphate (ATP) synthesis rate and tricarboxylic acid (TCA) cycle flux were reduced by 49% and 22%, respectively, in the cancer-bearing mice (p<0.008; t-test vs. controls). The ratio of ATP synthesis rate to the TCA cycle flux (an index of mitochondrial coupling) was reduced by 32% in the cancer-bearing mice (p=0.036; t-test vs. controls). Genomic analysis revealed aberrant expression levels for key regulatory genes and transmission electron microscopy (TEM) revealed ultrastructural abnormalities in the muscle fiber, consistent with the presence of abnormal, giant mitochondria. Taken together, these data suggest that mitochondrial uncoupling occurs in cancer cachexia and thus point to the mitochondria as a potential pharmaceutical target for the treatment of cachexia. These findings may prove relevant to elucidating the mechanisms underlying skeletal muscle wasting observed in other chronic diseases, as well as in aging.

  20. Uncoupling GP1 and GP2 expression in the Lassa virus glycoprotein complex: implications for GP1 ectodomain shedding

    Directory of Open Access Journals (Sweden)

    Illick Kerry A

    2008-12-01

    Full Text Available Abstract Background Sera from convalescent Lassa fever patients often contains antibodies to Lassa virus (LASV glycoprotein 1 (GP1, and glycoprotein 2 (GP2; Immunization of non-human primates with viral vectors expressing the arenaviral glycoprotein complex (GPC confers full protective immunity against a lethal challenge with LASV. Thus, the development of native or quasi native recombinant LASV GP1 and GP2 as soluble, uncoupled proteins will improve current diagnostics, treatment, and prevention of Lassa fever. To this end, mammalian expression systems were engineered for production and purification of secreted forms of soluble LASV GP1 and GP2 proteins. Results Determinants for mammalian cell expression of secreted uncoupled Lassa virus (LASV glycoprotein 1 (GP1 and glycoprotein 2 (GP2 were established. Soluble GP1 was generated using either the native glycoprotein precursor (GPC signal peptide (SP or human IgG signal sequences (s.s.. GP2 was secreted from cells only when (1 the transmembrane (TM domain was deleted, the intracellular domain (IC was fused to the ectodomain, and the gene was co-expressed with a complete GP1 gene in cis; (2 the TM and IC domains were deleted and GP1 was co-expressed in cis; (3 expression of GP1 was driven by the native GPC SP. These data implicate GP1 as a chaperone for processing and shuttling GP2 to the cell surface. The soluble forms of GP1 and GP2 generated through these studies were secreted as homogeneously glycosylated proteins that contained high mannose glycans. Furthermore, observation of GP1 ectodomain shedding from cells expressing wild type LASV GPC represents a novel aspect of arenaviral glycoprotein expression. Conclusion These results implicate GP1 as a chaperone for the correct processing and shuttling of GP2 to the cell surface, and suggest that native GPC SP plays a role in this process. In the absence of GP1 and GPC SP the GP2 protein may be processed by an alternate pathway that produces

  1. 77 FR 34783 - Highly Pathogenic Avian Influenza

    Science.gov (United States)

    2012-06-12

    ... Avian Influenza AGENCY: Animal and Plant Health Inspection Service, USDA. ACTION: Interim rule... importation of bird and poultry products from regions where any subtype of highly pathogenic avian influenza... avian influenza (HPAI). On January 24, 2011, we published in the Federal Register (76 FR...

  2. Markov Chain Estimation of Avian Seasonal Fecundity

    Science.gov (United States)

    To explore the consequences of modeling decisions on inference about avian seasonal fecundity we generalize previous Markov chain (MC) models of avian nest success to formulate two different MC models of avian seasonal fecundity that represent two different ways to model renestin...

  3. An overview on avian influenza

    Directory of Open Access Journals (Sweden)

    Nelson Rodrigo da Silva Martins

    2012-06-01

    Full Text Available Avian influenza (AI is considered an exotic disease in the Brazilian poultry industry, according to the National Avian Health Program (PNSA, with permanent monitoring of domestic, exotic and native avian species. Brazil presents privileged environmental conditions of reduced risk. In addition, all commercial poultry and conservation holdings are registered in state or national inventories and geographically located (GPS for health control. Poultry health standards are adopted for the conformity to the international market, mostly for the intensified poultry destined for exportation, but also for companion exotic and native conservation facilities. Guidelines for monitoring and the diagnosis of AI are published by the PNSA and follow the standards proposed by the international health code (World Organization for Animal Health, Organization International des Epizooties - OIE and insure the free of status for avian influenza virus (AIV of LPAIV-low pathogenicity AIV and HPAIV-high pathogenicity AIV. In addition, the infections by mesogenic and velogenic Newcastle disease virus, Mycoplasma gallisepticum, M. synoviae and M. meleagridis, Salmonella enteric subspecies enterica serovar Gallinarum biovars Gallinarum and Pullorum are eradicated from reproduction. Controlled infections by S.enterica subspecies enterica serovars Enteritidis and Typhimurium are monitored for breeders. The vaccination of chickens in ovo or at hatch against Marek's disease is mandatory. Broiler production is an indoor activity, confinement which insures biosecurity, with safe distances from the potential AIV reservoir avian species. Worldwide HPAIV H5N1 notifications to the OIE, in March 2011, included 51 countries.

  4. Avian influenza : a review article

    Directory of Open Access Journals (Sweden)

    A. Yalda

    2006-07-01

    Full Text Available The purpose of this paper is to provides general information about avian influenza (bird flu and specific information about one type of bird flu, called avian influenza A (H5N1, that has caused infections in birds in Asia and Europe and in human in Asia. The main materials in this report are based on the World Health Organization (WHO , world organization for animal health (OIE , food and agriculture organization of the united nations (FAO information and recommendations and review of the published literature about avian influenza. Since December 2003, highly pathogenic H5N1 avian influenza viruses have swept through poultry populations across Asia and parts of Europe. The outbreaks are historically unprecedented in scale and geographical spread. Their economic impact on the agricultural sector of the affected countries has been large. Human cases, with an overall fatality rate around 50%, have also been reported and almost all human infections can be linked to contact with infected poultry. Influenza viruses are genetically unstable and their behaviour cannot be predicted so the risk of further human cases persists. The human health implications have now gained importance, both for illness and fatalities that have occurred following natural infection with avian viruses, and for the potential of generating a re-assortant virus that could give rise to the next human influenza pandemic.

  5. Recombinant Expression and Biological Property Analysis of Goose Avian β-Defensin 7 Protein%重组鹅β-防御素7蛋白的原核表达及其生物学特性分析

    Institute of Scientific and Technical Information of China (English)

    高梦莹; 徐倩倩; 张婷婷; 陈玉秋; 赵文钧; 齐甜铭; 马得莹

    2016-01-01

    The objective of the study was to investigate the biological property of goose avian β-defensin ( AvBD) 7. The gene of AvBD7 was cloned into EcoRⅠ and XhoⅠ sites of pProEX HTa vector to construct recombinant plasmid pProEX-AvBD7. The recombinant plasmid was transformed into Escherichia coli Rosetta and the bacteria was induced with isopropy-β-D-thiogalactoside ( IPTG) . It was analyzed by tricine sodium do-decyl sulfate polyacrylamide gel electrophoresis ( Tricine-SDS-PAGE) that the recombinant protein size was 10 to 15 ku, which was the same as the expected size. Antimicrobial activity of the recombinant fusion protein a-gainst Escherichia coli, Salmonella pullorum, Staphylococcus aureus, Micrococcus tetragenus and Bacillus subtilis was measured in vitro. In addition, the effect of ionic strength on the antibacterial activity and hemolytic activity of the recombinant fusion protein were investigated. The results showed that the recombinant goose AvBD7 protein exhibited significant antimicrobial activity against the five bacteria investigated ( P0.05). In conclusion, recombinant goose AvBD7 protein exhibits extensive antimicrobial activity. High salt concentration significantly decrease its antibacterial activity. In addition, the recombinant goose AvBD7 protein shows little hemolytic activity.%为探讨鹅β-防御素7( AvBD7)的生物学特性,将鹅AvBD7基因亚克隆到大肠杆菌原核表达载体pProEX HTa的EcoRⅠ和XhoⅠ双酶切位点上,构建重组表达质粒pProEX-AvBD7,将重组质粒转化到大肠杆菌Rosetta感受态细胞中,用异丙基-β-D-硫代半乳糖苷( IPTG)对其菌液进行诱导表达。经 N-三(羟甲基)甲基甘氨酸十二烷基硫酸钠聚丙烯酰胺凝胶电泳( Tricine-SDS-PAGE)分析,该重组蛋白大小为10~15 ku,与预期大小结果一致。重组鹅AvBD7蛋白纯化后,通过菌落计数的方法测定其体外抗大肠杆菌、鸡白痢沙门氏菌、金黄色葡萄球菌、四联球菌、枯草芽孢杆

  6. Uncoupling of the LKB1-AMPKalpha energy sensor pathway by growth factors and oncogenic BRAF.

    Directory of Open Access Journals (Sweden)

    Rosaura Esteve-Puig

    Full Text Available BACKGROUND: Understanding the biochemical mechanisms contributing to melanoma development and progression is critical for therapeutical intervention. LKB1 is a multi-task Ser/Thr kinase that phosphorylates AMPK controlling cell growth and apoptosis under metabolic stress conditions. Additionally, LKB1(Ser428 becomes phosphorylated in a RAS-Erk1/2-p90(RSK pathway dependent manner. However, the connection between the RAS pathway and LKB1 is mostly unknown. METHODOLOGY/PRINCIPAL FINDINGS: Using the UV induced HGF transgenic mouse melanoma model to investigate the interplay among HGF signaling, RAS pathway and PI3K pathway in melanoma, we identified LKB1 as a protein directly modified by HGF induced signaling. A variety of molecular techniques and tissue culture revealed that LKB1(Ser428 (Ser431 in the mouse is constitutively phosphorylated in BRAF(V600E mutant melanoma cell lines and spontaneous mouse tumors with high RAS pathway activity. Interestingly, BRAF(V600E mutant melanoma cells showed a very limited response to metabolic stress mediated by the LKB1-AMPK-mTOR pathway. Here we show for the first time that RAS pathway activation including BRAF(V600E mutation promotes the uncoupling of AMPK from LKB1 by a mechanism that appears to be independent of LKB1(Ser428 phosphorylation. Notably, the inhibition of the RAS pathway in BRAF(V600E mutant melanoma cells recovered the complex formation and rescued the LKB1-AMPKalpha metabolic stress-induced response, increasing apoptosis in cooperation with the pro-apoptotic proteins Bad and Bim, and the down-regulation of Mcl-1. CONCLUSIONS/SIGNIFICANCE: These data demonstrate that growth factor treatment and in particular oncogenic BRAF(V600E induces the uncoupling of LKB1-AMPKalpha complexes providing at the same time a possible mechanism in cell proliferation that engages cell growth and cell division in response to mitogenic stimuli and resistance to low energy conditions in tumor cells. Importantly, this

  7. Avian Bornaviruses in North American Gulls.

    Science.gov (United States)

    Guo, Jianhua; Tizard, Ian; Baroch, John; Shivaprasad, H L; Payne, Susan L

    2015-07-01

    Avian bornaviruses, recently described members of the family Bornaviridae, have been isolated from captive parrots and passerines as well as wild waterfowl in which they may cause lethal neurologic disease. We report detection of avian bornavirus RNA in the brains of apparently healthy gulls. We tested 439 gull brain samples from 18 states, primarily in the northeastern US, using a reverse-transcriptase PCR assay with primers designed to detect a conserved region of the bornavirus M gene. Nine birds yielded a PCR product of appropriate size. Sequencing of PCR products indicated that the virus was closely related to aquatic bird bornavirus 1 (ABBV-1). Viral RNA was detected in Herring Gulls (Larus argentatus), Ring-billed Gulls (Larus delawarensis), and Laughing Gulls (Leucophaeus atricilla). Eight of the nine positive birds came from the New York/New Jersey area. One positive Herring Gull came from New Hampshire. Histopathologic examination of one well-preserved brain from a Herring Gull from Union County New Jersey, showed a lymphocytic encephalitis similar to that observed in bornavirus-infected parrots and geese. Bornavirus N protein was confirmed in two Herring Gull brains by immunohistochemistry. Thus ABBV-1 can infect gulls and cause encephalitic brain lesions similar to those observed in other birds.

  8. In vivo and in vitro effects of the mitochondrial uncoupler FCCP on microtubules.

    Science.gov (United States)

    Maro, B; Marty, M C; Bornens, M

    1982-01-01

    FCCP (carbonylcyanide-p-trifluoromethoxyphenylhydrazone), a potent uncoupler of oxidative phosphorylation, induces the complete disruption of cellular microtubules. A further analysis of this effect on BHK21 cells has shown that a decrease in the number of microtubules can be observed 15 min after adding FCCP and there is complete disruption after 60 min. Regrowth of microtubules was initiated 30 min after removal of FCCP, in marked contrast with the rapid reversion observed when microtubules are disrupted by nocodazole. A similar delay was required for the recovery of mitochondrial function as assessed by rhodamine 123 labelling. The effect of FCCP on microtubules was partially inhibited by preincubation of the cells with NaN3, suggesting that FCCP acts on microtubules through mitochondria. FCCP did not depolymerize microtubules of cells permeabilized with Triton X-100. In vitro polymerisation of microtubule protein was only slightly diminished by concentrations of FCCP which provoke complete disassembly in vivo. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the microtubules polymerized in vitro in the presence of FCCP showed a reduced amount of high mol. wt. proteins, mainly MAP 2, associated with them. In an attempt to reproduce the mitochondrial effects of FCCP in vitro, we checked the effects of alkaline pH and calcium on microtubule protein polymerization in the presence of FCCP. FCCP did not influence the calcium inhibitory effect but did significantly increase the inhibitory effect of alkaline pH. We conclude that FCCP could depolymerise microtubules in vivo through a dual operation: increasing the intracellular pH by the disruption of the mitochondrial H+ gradient and decreasing the stability of microtubules by impairing the binding of microtubule-associated proteins.

  9. Avian reproductive physiology

    Science.gov (United States)

    Gee, G.F.; Gibbons, Edward F.; Durrant, Barbara S.; Demarest, Jack

    1995-01-01

    Knowledge of the many physiological factors associated with egg production , fertility, incubation, and brooding in nondomestic birds is limited. Science knows even less about reproduction in most of the 238 endangered or threatened birds. This discussion uses studies of nondomestic and, when necessary, domestic birds to describe physiological control of reproduction. Studies of the few nondomestic avian species show large variation in physiological control of reproduction. Aviculturists, in order to successfully propagate an endangered bird, must understand the bird's reproductive peculiarities. First, investigators can do studies with carefully chosen surrogate species, but eventually they need to confirm the results in the target endangered bird. Studies of reproduction in nondomestic birds increased in the last decade. Still, scientists need to do more comparative studies to understand the mechanisms that control reproduction in birds. New technologies are making it possible to study reproductive physiology of nondomestic species in less limiting ways. These technologies include telemetry to collect information without inducing stress on captives (Howey et al., 1987; Klugman, 1987), new tests for most of the humoral factors associated with reproduction, and the skill to collect small samples and manipulate birds without disrupting the physiological mechanisms (Bercovitz et al., 1985). Managers are using knowledge from these studies to improve propagation in zoological parks, private and public propagation facilities, and research institutions. Researchers need to study the control of ovulation, egg formation, and oviposition in the species of nondomestic birds that lay very few eggs in a season, hold eggs in the oviduct for longer intervals, or differ in other ways from the more thoroughly studied domestic birds. Other techniques that would enhance propagation for nondomestlc birds include tissue culture of cloned embryonic cells, cryopreservation of embryos

  10. Molecular characterization of Indonesia avian influenza virus

    Directory of Open Access Journals (Sweden)

    N.L.P.I. Dharmayanti

    2005-06-01

    Full Text Available Avian influenza outbreaks in poultry have been reported in Java island since August 2003. A total of 14 isolates of avian influenza virus has been isolated from October 2003 to October 2004. The viruses have been identified as HPAI H5N1 subtype. All of them were characterized further at genetic level and also for their pathogenicity. Phylogenetic analysis showed all of the avian influenza virus isolates were closely related to avian influenza virus from China (A/Duck/China/E319-2/03(H5N1. Molecular basis of pathogenicity in HA cleavage site indicated that the isolates of avian influenza virus have multiple basic amino acid (B-X-B-R indicating that all of the isolates representing virulent avian influenza virus (highly pathogenic avian influenza virus.

  11. Uncoupling of Longevity and Telomere Length in C. elegans.

    Directory of Open Access Journals (Sweden)

    2005-09-01

    Full Text Available The nematode Caenorhabditis elegans, after completing its developmental stages and a brief reproductive period, spends the remainder of its adult life as an organism consisting exclusively of post-mitotic cells. Here we show that telomere length varies considerably in clonal populations of wild-type worms, and that these length differences are conserved over at least ten generations, suggesting a length regulation mechanism in cis. This observation is strengthened by the finding that the bulk telomere length in different worm strains varies considerably. Despite the close correlation of telomere length and clonal cellular senescence in mammalian cells, nematodes with long telomeres were neither long lived, nor did worm populations with comparably short telomeres exhibit a shorter life span. Conversely, long-lived daf-2 and short-lived daf-16 mutant animals can have either long or short telomeres. Telomere length of post-mitotic cells did not change during the aging process, and the response of animals to stress was found independent of telomere length. Collectively, our data indicate that telomere length and life span can be uncoupled in a post-mitotic setting, suggesting separate pathways for replication-dependent and -independent aging.

  12. Stochastic calculus for uncoupled continuous-time random walks.

    Science.gov (United States)

    Germano, Guido; Politi, Mauro; Scalas, Enrico; Schilling, René L

    2009-06-01

    The continuous-time random walk (CTRW) is a pure-jump stochastic process with several applications not only in physics but also in insurance, finance, and economics. A definition is given for a class of stochastic integrals driven by a CTRW, which includes the Itō and Stratonovich cases. An uncoupled CTRW with zero-mean jumps is a martingale. It is proved that, as a consequence of the martingale transform theorem, if the CTRW is a martingale, the Itō integral is a martingale too. It is shown how the definition of the stochastic integrals can be used to easily compute them by Monte Carlo simulation. The relations between a CTRW, its quadratic variation, its Stratonovich integral, and its Itō integral are highlighted by numerical calculations when the jumps in space of the CTRW have a symmetric Lévy alpha -stable distribution and its waiting times have a one-parameter Mittag-Leffler distribution. Remarkably, these distributions have fat tails and an unbounded quadratic variation. In the diffusive limit of vanishing scale parameters, the probability density of this kind of CTRW satisfies the space-time fractional diffusion equation (FDE) or more in general the fractional Fokker-Planck equation, which generalizes the standard diffusion equation, solved by the probability density of the Wiener process, and thus provides a phenomenologic model of anomalous diffusion. We also provide an analytic expression for the quadratic variation of the stochastic process described by the FDE and check it by Monte Carlo.

  13. Inhibiting avian influenza virus shedding using a novel RNAi antiviral vector technology: proof of concept in an avian cell model.

    Science.gov (United States)

    Linke, Lyndsey M; Wilusz, Jeffrey; Pabilonia, Kristy L; Fruehauf, Johannes; Magnuson, Roberta; Olea-Popelka, Francisco; Triantis, Joni; Landolt, Gabriele; Salman, Mo

    2016-03-01

    Influenza A viruses pose significant health and economic threats to humans and animals. Outbreaks of avian influenza virus (AIV) are a liability to the poultry industry and increase the risk for transmission to humans. There are limitations to using the AIV vaccine in poultry, creating barriers to controlling outbreaks and a need for alternative effective control measures. Application of RNA interference (RNAi) techniques hold potential; however, the delivery of RNAi-mediating agents is a well-known obstacle to harnessing its clinical application. We introduce a novel antiviral approach using bacterial vectors that target avian mucosal epithelial cells and deliver (small interfering RNA) siRNAs against two AIV genes, nucleoprotein (NP) and polymerase acidic protein (PA). Using a red fluorescent reporter, we first demonstrated vector delivery and intracellular expression in avian epithelial cells. Subsequently, we demonstrated significant reductions in AIV shedding when applying these anti-AIV vectors prophylactically. These antiviral vectors provided up to a 10,000-fold reduction in viral titers shed, demonstrating in vitro proof-of-concept for using these novel anti-AIV vectors to inhibit AIV shedding. Our results indicate this siRNA vector technology could represent a scalable and clinically applicable antiviral technology for avian and human influenza and a prototype for RNAi-based vectors against other viruses.

  14. Quantum Zeno Effect Underpinning the Radical-Ion-Pair Mechanism of Avian Magnetoreception

    CERN Document Server

    Kominis, I K

    2008-01-01

    The intricate biochemical processes underlying avian magnetoreception, the sensory ability of migratory birds to navigate using earths magnetic field, have been narrowed down to spin-dependent recombination of radical-ion pairs to be found in avian species retinal proteins. The avian magnetic field detection is governed by the interplay between magnetic interactions of the radicals unpaired electrons and the radicals recombination dynamics. Critical to this mechanism is the long lifetime of the radical-pair spin coherence, so that the weak geomagnetic field will have a chance to signal its presence. It is here shown that a fundamental quantum phenomenon, the quantum Zeno effect, is at the basis of the radical-ion-pair magnetoreception mechanism. The quantum Zeno effect naturally leads to long spin coherence lifetimes, without any constraints on the systems physical parameters, ensuring the robustness of this sensory mechanism. Basic experimental observations regarding avian magnetic sensitivity are seamlessly...

  15. Constitutive Mad1 targeting to kinetochores uncouples checkpoint signalling from chromosome biorientation.

    Science.gov (United States)

    Maldonado, Maria; Kapoor, Tarun M

    2011-04-01

    Accurate chromosome segregation depends on biorientation, whereby sister chromatids attach to microtubules from opposite spindle poles. The spindle-assembly checkpoint is a surveillance mechanism in eukaryotes that inhibits anaphase until all chromosomes have bioriented. In present models, the recruitment of the spindle-assembly checkpoint protein Mad2, through Mad1, to non-bioriented kinetochores is needed to stop cell-cycle progression. However, it is unknown whether Mad1-Mad2 targeting to kinetochores is sufficient to block anaphase. Furthermore, it is unclear whether regulators of biorientation (for example, Aurora kinases) have checkpoint functions downstream of Mad1-Mad2 recruitment or whether they act upstream to quench the primary error signal. Here, we engineered a Mad1 construct that localizes to bioriented kinetochores. We show that the kinetochore localization of Mad1 is sufficient for a metaphase arrest that depends on Mad1-Mad2 binding. By uncoupling the checkpoint from its primary error signal, we show that Aurora, Mps1 and BubR1 kinases, but not Polo-like kinase, are needed to maintain checkpoint arrest when Mad1 is present on kinetochores. Together, our data suggest a model in which the biorientation errors, which recruit Mad1-Mad2 to kinetochores, may be signalled not only through Mad2 template dynamics, but also through the activity of widely conserved kinases, to ensure the fidelity of cell division.

  16. Age-related changes of serum mitochondrial uncoupling 1, rumen and rectal temperature in goats.

    Science.gov (United States)

    Arfuso, Francesca; Rizzo, Maria; Giannetto, Claudia; Giudice, Elisabetta; Fazio, Francesco; Piccione, Giuseppe

    2016-07-01

    Thermoregulatory processes are induced not only by exposure to cold or heat but also by a variety of physiological situations including age, fasting and food intake that result in changes in body temperature. The aim of the present study was to evaluate the differences in serum mitochondrial uncoupling protein 1 (UCP1), rumen temperature (TRUMEN) and rectal temperature (TRECTAL) values between adult and kids goats. Ten adult male Maltese goats aged 3-5 years old (Group A) and 30 male kids, raised for meat, were enrolled in this study. The kids were equally divided into 3 groups according to their age: Group B included kids aged 3 months, Group C included kids aged 4 months and Group D included kids aged 5 months. Blood samples and measurements of TRUMEN and TRECTAL were obtained from each animal. One-way repeated measures analysis of variance (ANOVA) was applied to evaluate the effect of age on the studied parameters. Statistically significant higher serum UCP1 levels (Ptemperature suggesting that further details about the thermogenic capacity and the function of UCP1 in kids and adult goats are worth exploring.

  17. Uncoupling the functions of CALM in VAMP sorting and clathrin-coated pit formation.

    Directory of Open Access Journals (Sweden)

    Daniela A Sahlender

    Full Text Available CALM (clathrin assembly lymphoid myeloid leukemia protein is a cargo-selective adaptor for the post-Golgi R-SNAREs VAMPs 2, 3, and 8, and it also regulates the size of clathrin-coated pits and vesicles at the plasma membrane. The present study has two objectives: to determine whether CALM can sort additional VAMPs, and to investigate whether VAMP sorting contributes to CALM-dependent vesicle size regulation. Using a flow cytometry-based endocytosis efficiency assay, we demonstrate that CALM is also able to sort VAMPs 4 and 7, even though they have sorting signals for other clathrin adaptors. CALM homologues are present in nearly every eukaryote, suggesting that the CALM family may have evolved as adaptors for retrieving all post-Golgi VAMPs from the plasma membrane. Using a knockdown/rescue system, we show that wild-type CALM restores normal VAMP sorting in CALM-depleted cells, but that two non-VAMP-binding mutants do not. However, when we assayed the effect of CALM depletion on coated pit morphology, using a fluorescence microscopy-based assay, we found that the two mutants were as effective as wild-type CALM. Thus, we can uncouple the sorting function of CALM from its structural role.

  18. Uncoupling the functions of CALM in VAMP sorting and clathrin-coated pit formation.

    Science.gov (United States)

    Sahlender, Daniela A; Kozik, Patrycja; Miller, Sharon E; Peden, Andrew A; Robinson, Margaret S

    2013-01-01

    CALM (clathrin assembly lymphoid myeloid leukemia protein) is a cargo-selective adaptor for the post-Golgi R-SNAREs VAMPs 2, 3, and 8, and it also regulates the size of clathrin-coated pits and vesicles at the plasma membrane. The present study has two objectives: to determine whether CALM can sort additional VAMPs, and to investigate whether VAMP sorting contributes to CALM-dependent vesicle size regulation. Using a flow cytometry-based endocytosis efficiency assay, we demonstrate that CALM is also able to sort VAMPs 4 and 7, even though they have sorting signals for other clathrin adaptors. CALM homologues are present in nearly every eukaryote, suggesting that the CALM family may have evolved as adaptors for retrieving all post-Golgi VAMPs from the plasma membrane. Using a knockdown/rescue system, we show that wild-type CALM restores normal VAMP sorting in CALM-depleted cells, but that two non-VAMP-binding mutants do not. However, when we assayed the effect of CALM depletion on coated pit morphology, using a fluorescence microscopy-based assay, we found that the two mutants were as effective as wild-type CALM. Thus, we can uncouple the sorting function of CALM from its structural role.

  19. Evolution of Avian Tumor Viruses

    Science.gov (United States)

    Virus-induced neoplastic diseases of poultry, namely Marek’s disease (MD), induced by a herpesvirus, and the avian leukosis and reticuloendotheliosis induced by retroviruses, can cause significant economic losses from tumor mortality as well as poor performance. Successful control of MD is and has ...

  20. Avian Paramyxovirus: A Brief Review.

    Science.gov (United States)

    Gogoi, P; Ganar, K; Kumar, S

    2017-02-01

    Avian paramyxoviruses (APMVs) have been reported from a wide variety of avian species around the world. Avian paramyxoviruses are economically significant because of the huge mortality and morbidity associated with it. Twelve different serotypes of APMV have been reported till date. Avian paramyxoviruses belong to the family Paramyxoviridae under genus Avulavirus. Newcastle disease virus (APMV-1) is the most characterized members among the APMV serotypes. Complete genome sequence of all twelve APMV serotypes has been published recently. In recent years, APMV-1 has attracted the virologists for its oncolytic activity and its use as a vaccine vector for both animals and humans. The recombinant APMV-based vaccine offers a pertinent choice for the construction of live attenuated vaccine due to its minimum recombination frequency, modular nature of transcription and lack of DNA phase during its replication. Although insufficient data are available regarding other APMV serotypes, our understanding about the APMV biology is expanding rapidly because of the availability of modern molecular biology tools and high-throughput complete genome sequencing.

  1. Development of an antigen-capture ELISA for the detection of avian leukosis virus p27 antigen.

    Science.gov (United States)

    Yun, Bingling; Li, Delong; Zhu, Haibo; Liu, Wen; Qin, Liting; Liu, Zaisi; Wu, Guan; Wang, Yongqiang; Qi, Xiaole; Gao, Honglei; Wang, Xiaomei; Gao, Yulong

    2013-02-01

    An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) employing monoclonal and polyclonal antibodies against p27 was developed for the detection of the avian leukosis virus (ALV). The specificity of the optimized AC-ELISA was evaluated using avian leukosis virus subgroup J (ALV-J), avian leukosis virus subgroup A (ALV-A), avian leukosis virus subgroup B (ALV-B), avian infectious bronchitis virus (IBV), Marek's disease virus (MDV), avian infectious laryngotracheitis virus (ILTV), Fowlpox virus (FPV), infectious bursal disease virus (IBDV), Newcastle disease virus (NDV), avian reovirus (ARV), reticuloendotheliosis virus (REV), avian influenza virus (AIV) and Escherichia coli. The only specimens that yielded a strong signal were ALV-J, ALV-A and ALV-B, indicating that this assay is suitable for the detection of ALV. The limit of detection of this assay was 1.25 ng/ml of rp27 protein and 10(1.79)TCID(50) units of HLJ09MDJ-1 (ALV-J). Moreover, this AC-ELISA can detect ALV in cloacal swabs of chickens experimentally infected as early as 12 days post-infection. The AC-ELISA detected the virus in the albumin and cloacal swabs of naturally infected chickens, and the results were confirmed by PCR, indicating that the AC-ELISA was a suitable method for the detection of ALV. This test is rapid and sensitive and could be convenient for epidemiological studies and eradication programs.

  2. Tissue interactions of avian viral attachment proteins

    NARCIS (Netherlands)

    Ambepitiya Wickramasinghe, I.N.

    2015-01-01

    Viruses can infect a wide range of hosts; varying from bacteria and plants to animals and humans. While many viral infections may pass unnoticed, some are of major importance due to their implications on health and welfare of plants, animals and/or humans. In particular, viruses that can infect avia

  3. Avian disease at the Salton Sea

    Science.gov (United States)

    Friend, M.

    2002-01-01

    A review of existing records and the scientific literature was conducted for occurrences of avian diseases affecting free-ranging avifauna within the Salton Sea ecosystem. The period for evaluation was 1907 through 1999. Records of the U.S. Department of Agriculture, Bureau of Biological Survey and the scientific literature were the data sources for the period of 1907a??1939. The narrative reports of the U.S. Fish and Wildlife Service's Sonny Bono National Wildlife Refuge Complex and the epizootic database of the U.S. Geological Survey's National Wildlife Health Center were the primary data sources for the remainder of the evaluation. The pattern of avian disease at the Salton Sea has changed greatly over time. Relative to past decades, there was a greater frequency of major outbreaks of avian disease at the Salton Sea during the 1990s than in previous decades, a greater variety of disease agents causing epizootics, and apparent chronic increases in the attrition of birds from disease. Avian mortality was high for about a decade beginning during the mid-1920s, diminished substantially by the 1940s and was at low to moderate levels until the 1990s when it reached the highest levels reported. Avian botulism (Clostridium botulinum type C) was the only major cause of avian disease until 1979 when the first major epizootic of avian cholera (Pasteurella multocidia) was documented. Waterfowl and shorebirds were the primary species affected by avian botulism. A broader spectrum of species have been killed by avian cholera but waterfowl have suffered the greatest losses. Avian cholera reappeared in 1983 and has joined avian botulism as a recurring cause of avian mortality. In 1989, avian salmonellosis (Salmonella typhimurium) was first diagnosed as a major cause of avian disease within the Salton Sea ecosystem and has since reappeared several times, primarily among cattle egrets (Bubulcus ibis). The largest loss from a single epizootic occurred in 1992, when an estimated

  4. Effects of endothelial nitric oxide synthase uncoupling on pulmonary endothelial dysfunction in rats with decompression sickness

    Institute of Scientific and Technical Information of China (English)

    Hai-Shan Lin; Min Ou; Yi-Qun Fang

    2015-01-01

    Background:To investigate the effects of unsafe decompression on rat pulmonary endothelial function and its relevant mechanisms. Methods: Sixty male Sprague-Dawley (SD) rats were randomly divided into a control group (n=30) and a decompression sickness (DCS) group (n=30). The DCS model was established by placing the rats in the DCS group in a pressurized cabin where they were exposed to a 600 kPa compressed air environment for 60 min, and the pressure was then reduced by 100 kPa/min until it reached atmospheric pressure. After the surviving rats in the DCS group and the rats in the control group were anesthetized, their pulmonary arteries were stripped to test the in vitro pulmonary artery endothelium-dependent vasodilation capacity. Western blotting was used to measure the expression and dissociation of endothelial nitric oxide synthase (eNOS) in pulmonary artery tissues and all protein nitration levels in pulmonary artery tissues; reactive oxygen species (ROS) formation was measured via in vitro pulmonary artery superoxide anion probe dihydroethidium (DHE) staining. Results: After experiencing unsafe decompression, 10 of the 30 rats in the DCS group died. The pulmonary artery endothelium-dependent vasodilation capacity in the surviving rats decreased significantly (P0.05), but the ratio of eNOS monomer/dimer in the DCS group was significantly higher than that in the control group (P Conclusion: Unsafe decompression during a simulated submarine escape process can lead to eNOS dimer uncoupling in the pulmonary artery endothelium. The dissociated eNOS monomer cannot synthesize nitric oxide (NO) and thus affect the endothelium-dependent vasodilation capacity. The eNOS monomer can promote peroxynitrite (ONOO–) synthesis, leading to an increase in protein tyrosine nitration levels in pulmonary artery tissues and causing disorder in cell cycle regulation. The eNOS monomer can also cause an increase in the formation of ROS and thus mediate peroxidation damage.

  5. Characterization of avian influenza H5N1 virosome

    Directory of Open Access Journals (Sweden)

    Chatchai Sarachai

    2014-04-01

    Full Text Available The purpose of this study was to prepare and characterize virosome containing envelope proteins of the avian influenza (H5N1 virus. The virosome was prepared by the solubilization of virus with octaethyleneglycol mono (n-dodecyl ether (C12E8 followed by detergent removal with SM2 Bio-Beads. Biochemical analysis by SDS-PAGE and western blotting, indicated that avian influenza H5N1 virosome had similar characteristics to the parent virus and contained both the hemagglutinin (HA, 60-75 kDa and neuraminidase (NA, 220 kDa protein, with preserved biological activity, such as hemagglutination activity. The virosome structure was analyzed by negative stained transmission electron microscope (TEM demonstrated that the spherical shapes of vesicles with surface glycoprotein spikes were harbored. In conclusion, the biophysical properties of the virosome were similar to the parent virus, and the use of octaethyleneglycol mono (n-dodecyl ether to solubilize viral membrane, followed by removal of detergent using polymer beads adsorption (Bio-Beads SM2 was the preferable method for obtaining avian influenza virosome. The outcome of this study might be useful for further development veterinary virus vaccines.

  6. Development and application of an indirect ELISA for detection of antibodies against avian hepatitis E virus.

    Science.gov (United States)

    Zhao, Qin; Sun, Yani; Zhao, Jinan; Hu, Shoubin; Zhao, Feifei; Chen, Fuyong; Clavijo, Alfonso; Zhou, En-Min; Xiao, Yihong

    2013-01-01

    An indirect enzyme-linked immunosorbent assay (iELISA) that could detect immunoglobulin G antibodies against avian hepatitis E virus (HEV) was developed. This assay employs a truncated C-terminal 268-amino acid recombinant ORF2 protein from an avian HEV genotype 3 strain isolated in China (CaHEV) as the coating antigen. The antigen concentration and serum dilution were optimized using a checkerboard titration. A cut-off value of 0.368 at OD(450nm) was determined by testing 120 positive and 200 negative chicken sera for avian HEV antibodies using the two-graph receiver operating characteristic (TG-ROC) analysis. This iELISA has a sensitivity of 96.1% and a specificity of 95.8%. The overall agreement between the iELISA and a corresponding Western blot was 97%. The iELISA was used to evaluate the seroprevalence of avian HEV in poultry farms in the Shandong province. The avian HEV seropositive rate of 35.9% was determined by testing 1871 serum samples that were collected from 10 chicken flocks ranged from 10 to 60 weeks of age. The iELISA that was developed in this study can be used for detection of immunoglobulin G antibodies against avian HEV.

  7. Thermodynamics of Anharmonic Systems: Uncoupled Mode Approximations for Molecules.

    Science.gov (United States)

    Li, Yi-Pei; Bell, Alexis T; Head-Gordon, Martin

    2016-06-14

    The partition functions, heat capacities, entropies, and enthalpies of selected molecules were calculated using uncoupled mode (UM) approximations, where the full-dimensional potential energy surface for internal motions was modeled as a sum of independent one-dimensional potentials for each mode. The computational cost of such approaches scales the same with molecular size as standard harmonic oscillator vibrational analysis using harmonic frequencies (HO(hf)). To compute thermodynamic properties, a computational protocol for obtaining the energy levels of each mode was established. The accuracy of the UM approximation depends strongly on how the one-dimensional potentials of each modes are defined. If the potentials are determined by the energy as a function of displacement along each normal mode (UM-N), the accuracies of the calculated thermodynamic properties are not significantly improved versus the HO(hf) model. Significant improvements can be achieved by constructing potentials for internal rotations and vibrations using the energy surfaces along the torsional coordinates and the remaining vibrational normal modes, respectively (UM-VT). For hydrogen peroxide and its isotopologs at 300 K, UM-VT captures more than 70% of the partition functions on average. By contrast, the HO(hf) model and UM-N can capture no more than 50%. For a selected test set of C2 to C8 linear and branched alkanes and species with different moieties, the enthalpies calculated using the HO(hf) model, UM-N, and UM-VT are all quite accurate comparing with reference values though the RMS errors of the HO model and UM-N are slightly higher than UM-VT. However, the accuracies in entropy calculations differ significantly between these three models. For the same test set, the RMS error of the standard entropies calculated by UM-VT is 2.18 cal mol(-1) K(-1) at 1000 K. By contrast, the RMS error obtained using the HO model and UM-N are 6.42 and 5.73 cal mol(-1) K(-1), respectively. For a test set

  8. Avian malaria in New Zealand.

    Science.gov (United States)

    Schoener, E R; Banda, M; Howe, L; Castro, I C; Alley, M R

    2014-07-01

    Avian malaria parasites of the genus Plasmodium have the ability to cause morbidity and mortality in naïve hosts, and their impact on the native biodiversity is potentially serious. Over the last decade, avian malaria has aroused increasing interest as an emerging disease in New Zealand with some endemic avian species, such as the endangered mohua (Mohua ochrocephala), thought to be particularly susceptible. To date, avian malaria parasites have been found in 35 different bird species in New Zealand and have been diagnosed as causing death in threatened species such as dotterel (Charadrius obscurus), South Island saddleback (Philesturnus carunculatus carunculatus), mohua, hihi (Notiomystis cincta) and two species of kiwi (Apteryx spp.). Introduced blackbirds (Turdus merula) have been found to be carriers of at least three strains of Plasmodium spp. and because they are very commonly infected, they are likely sources of infection for many of New Zealand's endemic birds. The spread and abundance of introduced and endemic mosquitoes as the result of climate change is also likely to be an important factor in the high prevalence of infection in some regions and at certain times of the year. Although still limited, there is a growing understanding of the ecology and epidemiology of Plasmodium spp. in New Zealand. Molecular biology has played an important part in this process and has markedly improved our understanding of the taxonomy of the genus Plasmodium. This review presents our current state of knowledge, discusses the possible infection and disease outcomes, the implications for host behaviour and reproduction, methods of diagnosis of infection, and the possible vectors for transmission of the disease in New Zealand.

  9. Using EGEE against avian flu

    CERN Multimedia

    2006-01-01

    During April 2006 avian flu was spreading across the world with the potential of turning into a pandemic, a drug to treat the deadly H5N1 strain was needed. Such a task required the huge processing power provided by EGEE, which analysed 300 000 possible drug components for their suitability. This map shows the network of computer centres and their activity during this time.

  10. Gender determination of avian embryo

    Science.gov (United States)

    Daum, Keith A.; Atkinson, David A.

    2002-01-01

    Disclosed is a method for gender determination of avian embryos. During the embryo incubation process, the outer hard shells of eggs are drilled and samples of allantoic fluid are removed. The allantoic fluids are directly introduced into an ion mobility spectrometer (IMS) for analysis. The resulting spectra contain the relevant marker peaks in the positive or negative mode which correlate with unique mobilities which are sex-specific. This way, the gender of the embryo can be determined.

  11. Avian zoonoses – a review

    Directory of Open Access Journals (Sweden)

    Kozdruń Wojciech

    2015-06-01

    Full Text Available Birds are one of the most interesting and most colourful groups of animals, but they can also be a source of zoonotic factors dangerous for humans. This paper describes the threats to human health from contact with birds. The most vulnerable occupational groups associated with birds are veterinarians, owners of poultry farms, breeders of ornamental birds, zoo personnel, and poultry slaughterhouse workers. Ornithosis is the most dangerous zoonosis of the avian bacterial diseases. Among other hazardous bacterial factors, Salmonella and Campylobacter are responsible for gastrointestinal diseases. Avian influenza is the most dangerous of the viral diseases. It should be noted, however, that avian influenza is a disease of birds, not humans. The recent threat which has appeared is infection with West Nile virus. The results of serological examinations of birds and humans indicate that the virus exists in our ecosystem. Allergic alveolitis connected with the pigeon tick and the Dermanyssus gallinae mite also merits mention. In any case, where people have contact with birds or their droppings and secretions, special precautions should be taken. This way the negative effects of birds on human health can be minimised or eliminated

  12. Lemna (duckweed) expressed hemagglutinin from avian influenza H5N1 protects chickens against H5N1 high pathogenicity avian influenza virus challenge

    Science.gov (United States)

    In the last two decades, transgenic plants have been explored as safe and cost effective alternative expression platforms for producing recombinant proteins. In this study, a synthetic hemagglutinin (HA) gene from the high pathogenicity avian influenza (HPAI) virus A/chicken/Indonesia/7/2003 (H5N1)...

  13. Effect of 6-ketocholestanol on FCCP- and DNP-induced uncoupling in plant mitochondria.

    Science.gov (United States)

    Vianello, A; Macri, F; Braidot, E; Mokhova, E N

    1995-05-22

    Effect of 6-ketocholestanol on FCCP-induced and DNP-induced uncoupling in beef liver and pea stem mitochondria was studied, under experimental conditions at which this steroid abolished the effect of low concentrations of FCCP and other most potent uncouplers in rat mitochondria [Starkov et al. (1994) FEBS Lett., 355, 305-308]. It is shown that, in both types of mitochondria, 6-ketocholestanol prevents or reverses the uncoupling induced by low concentrations of FCCP, but not that caused by high concentrations of FCCP or by any concentration of DNP. Progesterone and male sex hormones, showing recoupling capability in animal mitochondria, appear to be ineffective in the plant system. Cholesterol does not recouple in both animal and plant mitochondria. Plant steroids, such as beta-sitosterol and stigmasterol, are also without effect.

  14. Rem uncouples excitation–contraction coupling in adult skeletal muscle fibers

    Science.gov (United States)

    Beqollari, Donald; Romberg, Christin F.; Filipova, Dilyana; Meza, Ulises; Papadopoulos, Symeon

    2015-01-01

    In skeletal muscle, excitation–contraction (EC) coupling requires depolarization-induced conformational rearrangements in L-type Ca2+ channel (CaV1.1) to be communicated to the type 1 ryanodine-sensitive Ca2+ release channel (RYR1) of the sarcoplasmic reticulum (SR) via transient protein–protein interactions. Although the molecular mechanism that underlies conformational coupling between CaV1.1 and RYR1 has been investigated intensely for more than 25 years, the question of whether such signaling occurs via a direct interaction between the principal, voltage-sensing α1S subunit of CaV1.1 and RYR1 or through an intermediary protein persists. A substantial body of evidence supports the idea that the auxiliary β1a subunit of CaV1.1 is a conduit for this intermolecular communication. However, a direct role for β1a has been difficult to test because β1a serves two other functions that are prerequisite for conformational coupling between CaV1.1 and RYR1. Specifically, β1a promotes efficient membrane expression of CaV1.1 and facilitates the tetradic ultrastructural arrangement of CaV1.1 channels within plasma membrane–SR junctions. In this paper, we demonstrate that overexpression of the RGK protein Rem, an established β subunit–interacting protein, in adult mouse flexor digitorum brevis fibers markedly reduces voltage-induced myoplasmic Ca2+ transients without greatly affecting CaV1.1 targeting, intramembrane gating charge movement, or releasable SR Ca2+ store content. In contrast, a β1a-binding–deficient Rem triple mutant (R200A/L227A/H229A) has little effect on myoplasmic Ca2+ release in response to membrane depolarization. Thus, Rem effectively uncouples the voltage sensors of CaV1.1 from RYR1-mediated SR Ca2+ release via its ability to interact with β1a. Our findings reveal Rem-expressing adult muscle as an experimental system that may prove useful in the definition of the precise role of the β1a subunit in skeletal-type EC coupling. PMID:26078055

  15. Structural differences between the avian and human H7N9 hemagglutinin proteins are attributable to modifications in salt bridge formation: a computational study with implications in viral evolution.

    Directory of Open Access Journals (Sweden)

    Marni E Cueno

    Full Text Available Influenza A hemagglutinin (HA is a homotrimeric glycoprotein composed of a fibrous globular stem supporting a globular head containing three sialic acid binding sites responsible for infection. The H7N9 strain has consistently infected an avian host, however, the novel 2013 strain is now capable of infecting a human host which would imply that the HA in both strains structurally differ. A better understanding of the structural differences between the avian and human H7N9 strains may shed light into viral evolution and transmissibility. In this study, we elucidated the structural differences between the avian and human H7N9 strains. Throughout the study, we generated HA homology models, verified the quality of each model, superimposed HA homology models to determine structural differences, and, likewise, elucidated the probable cause for these structural differences. We detected two different types of structural differences between the novel H7N9 human and representative avian strains, wherein, one type (Pattern-1 showed three non-overlapping regions while the other type (Pattern-2 showed only one non-overlapping region. In addition, we found that superimposed HA homology models exhibiting Pattern-1 contain three non-overlapping regions designated as: Region-1 (S1571-A1601; Region-3 (R2621-S2651; and Region-4 (S2701-D2811, whereas, superimposed HA homology models showing Pattern-2 only contain one non-overlapping region designated as Region-2 (S1371-S1451. We attributed the two patterns we observed to either the presence of salt bridges involving the E1141 residue or absence of the R1411:D771 salt bridge. Interestingly, comparison between the human H7N7 and H7N9 HA homology models showed high structural similarity. We propose that the putative absence of the R1411:D771 salt bridge coupled with the putative presence of the E1141:R2621 and E1141:K2641 salt bridges found in the 2013 H7N9 HA homology model is associated to human-type receptor binding

  16. 禽白血病病毒p27蛋白在大肠杆菌中的表达和多克隆抗体的制备%Expression of p27 Protein of Avian Leukosis Virus in Escherichia coli and Preparation of Its Polyclonal Antibody

    Institute of Scientific and Technical Information of China (English)

    毛娅卿; 王嘉; 吴涛; 王哲; 蒋桃珍

    2014-01-01

    p27 protein gene from avian leukosis virus was cloned and constructed with plasmid pET-28a(+) . The recombinant protein p27 was expressed in an E. coli strain BL21(DE3) with IPTG induction and detected by Western blot with horseradish peroxidase labeled rabbit-anti-p27 antibodies. After that,the protein was purified to above 95% of purity by affinity chromatography with Ni-NTA agarose column. The anti-sera against p27 protein was obtained by immunizing rabbit with the purified recombinant p27 protein. The specificity of polyclonal antibody was about 1 ∶ 256000 after detected by ELISA. The results show that the p27 protein expressed by E. coli has good antigenicity and can replace the purified virus protein for detection of avian leukosis virus.%将禽白血病病毒p27基因克隆并构建重组表达质粒pET28a-p27,在大肠杆菌BL21中经IP TG诱导后产生可溶性表达蛋白。表达的蛋白用Western blot进行活性检测,其能与辣根过氧化物酶标记的兔抗p27抗体发生特异性反应;采用金属螯和层析对表达蛋白进行纯化,其纯度约为95%;用纯化的表达蛋白p27免疫家兔制备抗血清,ELISA抗体效价可达1∶25600。研究结果表明,大肠杆菌表达的p27蛋白具有良好的抗原性,可以替代纯化病毒蛋白用于禽白血病病毒检测。

  17. Mechanisms of avian songs and calls

    DEFF Research Database (Denmark)

    Larsen, Ole Næsbye

    2008-01-01

    The avian vocal organ, the syrinx, is a specialized structure located rather inaccessibly in an air sac close to the heart where the trachea bifurcates into the two primary bronchi. The syrinx of different avian taxa varies so much in position and morphology that it has been used for taxonomy. It...

  18. BCNU-induced gR2 defect mediates S-glutathionylation of Complex I and respiratory uncoupling in myocardium.

    Science.gov (United States)

    Kang, Patrick T; Chen, Chwen-Lih; Ren, Pei; Guarini, Giacinta; Chen, Yeong-Renn

    2014-06-15

    A deficiency of mitochondrial glutathione reductase (or GR2) is capable of adversely affecting the reduction of GSSG and increasing mitochondrial oxidative stress. BCNU [1,3-bis (2-chloroethyl)-1-nitrosourea] is an anticancer agent and known inhibitor of cytosolic GR ex vivo and in vivo. Here we tested the hypothesis that a BCNU-induced GR2 defect contributes to mitochondrial dysfunction and subsequent impairment of heart function. Intraperitoneal administration of BCNU (40 mg/kg) specifically inhibited GR2 activity by 79.8 ± 2.7% in the mitochondria of rat heart. However, BCNU treatment modestly enhanced the activities of mitochondrial Complex I and other ETC components. The cardiac function of BCNU-treated rats was analyzed by echocardiography, revealing a systolic dysfunction associated with decreased ejection fraction, decreased cardiac output, and an increase in left ventricular internal dimension and left ventricular volume in systole. The respiratory control index of isolated mitochondria from the myocardium was moderately decreased after BCNU treatment, whereas NADH-linked uncoupling of oxygen consumption was significantly enhanced. Extracellular flux analysis to measure the fatty acid oxidation of myocytes indicated a 20% enhancement after BCNU treatment. When the mitochondria were immunoblotted with antibodies against GSH and UCP3, both protein S-glutathionylation of Complex I and expression of UCP3 were significantly up-regulated. Overexpression of SOD2 in the myocardium significantly reversed BCNU-induced GR2 inhibition and mitochondrial impairment. In conclusion, BCNU-mediated cardiotoxicity is characterized by the GR2 deficiency that negatively regulates heart function by impairing mitochondrial integrity, increasing oxidative stress with Complex I S-glutathionylation, and enhancing uncoupling of mitochondrial respiration.

  19. Functional characterization of UCP1 in mammalian HEK293 cells excludes mitochondrial uncoupling artefacts and reveals no contribution to basal proton leak.

    Science.gov (United States)

    Jastroch, Martin; Hirschberg, Verena; Klingenspor, Martin

    2012-09-01

    Mechanistic studies on uncoupling proteins (UCPs) not only are important to identify their cellular function but also are pivotal to identify potential drug targets to manipulate mitochondrial energy transduction. So far, functional and comparative studies of uncoupling proteins in their native environment are hampered by different mitochondrial, cellular and genetic backgrounds. Artificial systems such as yeast ectopically expressing UCPs or liposomes with reconstituted UCPs were employed to address crucial mechanistic questions but these systems also produced inconsistencies with results from native mitochondria. We here introduce a novel mammalian cell culture system (Human Embryonic Kidney 293 - HEK293) to study UCP1 function. Stably transfected HEK293 cell lines were derived that contain mouse UCP1 at concentrations comparable to tissue mitochondria. In this cell-based test system UCP1 displays native functional behaviour as it can be activated with fatty acids (palmitate) and inhibited with purine nucleotides guanosine-diphosphate (GDP). The catalytic centre activity of the UCP1 homodimer in HEK293 is comparable to activities in brown adipose tissue supporting functionality of UCP1. Importantly, at higher protein levels than in yeast mitochondria, UCP1 in HEK293 cell mitochondria is fully inhibitable and does not contribute to basal proton conductance, thereby emphasizing the requirement of UCP1 activation for therapeutic purposes. These findings and resulting analysis on UCP1 characteristics demonstrate that the mammalian HEK293 cell system is suitable for mechanistic and comparative functional studies on UCPs and provides a non-confounding mitochondrial, cellular and genetic background.

  20. Avian bornavirus in the urine of infected birds

    Directory of Open Access Journals (Sweden)

    Villalobos AR

    2012-06-01

    Full Text Available J Jill Heatley,1 Alice R Villalobos21Zoological Medicine, 2Department of Nutrition & Food Science, Texas A&M University, College of Veterinary Medicine and Biomedical Sciences, College Station, TX, USAAbstract: Avian bornavirus (ABV causes proventricular dilatation disease in multiple avian species. In severe clinical disease, the virus, while primarily neurotropic, can be detected in many organs, including the kidneys. We postulated that ABV could be shed by the kidneys and found in the urine of infected birds. Immunohistochemical staining demonstrated viral N and P proteins of ABV within the renal tubules. We adapted a nonsurgical method of urine collection for use in parrots known to be shedding ABV in their droppings. We obtained urine without feces, and results were compared with swabs of fresh voided feces. Reverse transcription–polymerase chain reaction assay performed on these paired samples from five birds indicated that ABV was shed in quantity in the urine of infected birds, and a single sample was urine-positive and fecal-negative. We suggest that urine sampling may be a superior sample for detection of birds shedding ABV, and advocate that additional birds, known to be shedding or infected with ABV, should be investigated via this method.Keywords: avian bornavirus, Psittaciformes, parrot, urine, proventricular dilatation disease

  1. Analysis of Avian Hepatitis E Virus from Chickens, China

    OpenAIRE

    Zhao, Qin; Zhou, En Min; Dong, Shi Wei; Qiu, Hong Kai; Zhang, Lu; Hu, Shou Bin; Zhao, Fei Fei; Jiang, Shi Jin; Sun, Ya Ni

    2010-01-01

    Avian hepatitis E virus (HEV) has been identified in chickens; however, only 4 complete or near-complete genomic sequences have been reported. We found that the near-complete genomic sequence of avian HEV in chickens from China shared the highest identity (98.3%) with avian HEV from Europe and belonged to avian HEV genotype 3.

  2. Analysis of avian hepatitis E virus from chickens, China.

    Science.gov (United States)

    Zhao, Qin; Zhou, En Min; Dong, Shi Wei; Qiu, Hong Kai; Zhang, Lu; Hu, Shou Bin; Zhao, Fei Fei; Jiang, Shi Jin; Sun, Ya Ni

    2010-09-01

    Avian hepatitis E virus (HEV) has been identified in chickens; however, only 4 complete or near-complete genomic sequences have been reported. We found that the near-complete genomic sequence of avian HEV in chickens from China shared the highest identity (98.3%) with avian HEV from Europe and belonged to avian HEV genotype 3.

  3. Synergistic Effect of S224P and N383D Substitutions in the PA of H5N1 Avian Influenza Virus Contributes to Mammalian Adaptation.

    Science.gov (United States)

    Song, Jiasheng; Xu, Jing; Shi, Jianzhong; Li, Yanbing; Chen, Hualan

    2015-05-22

    The adaptation of H5N1 avian influenza viruses to human poses a great threat to public health. Previous studies indicate the adaptive mutations in viral polymerase of avian influenza viruses are major contributors in overcoming the host species barrier, with the majority of mammalian adaptive mutations occurring in the PB2 protein. However, the adaptive mutations in the PA protein of the H5N1 avian influenza virus are less defined and poorly understood. In this study, we identified the synergistic effect of the PA/224P + 383D of H5N1 avian influenza viruses and its ability to enhance the pathogenicity and viral replication in a mammalian mouse model. Interestingly, the signature of PA/224P + 383D mainly exists in mammalian isolates of the H5N1 influenza virus and pdmH1N1 influenza virus, providing a potential pathway for the natural adaptation to mammals which imply the effects of natural adaptation to mammals. Notably, the mutation of PA/383D, which is highly conserved in avian influenza viruses, increases the polymerase activity in both avian and human cells, and may have roles in maintaining the avian influenza virus in their avian reservoirs, and jumping species to infect humans.

  4. Immunoselection of cDNAs to avian intestinal calcium binding protein 28K and a novel calmodulin-like protein: assessment of mRNA regulation by the Vitamin D hormone

    Energy Technology Data Exchange (ETDEWEB)

    Mangelsdorf, D.J.; Komm, B.S.; McDonnell, D.P.; Pike, J.W.; Haussler, M.R.

    1987-12-15

    Calcium's role in a variety of cellular processes has been well documented. The storage, distribution, and delivery of calcium are regulated by a family of binding proteins including troponin C, calmodulin, parvalbumin, and vitamin D dependent calcium binding protein (CaBP-28), all of which have evolved from a common ancestral gene. To evaluate vitamin D regulation of gene transcription, a CaBP-28 cDNA (767 base pairs) was isolated from a chicken intestine lambdagt11 library utilizing a polyvalent CaBP-28 antibody as a probe. Coincident with the identification of the CaBP-28 cDNA, a group of cDNAs also was isolated (with the anti-CaBP-28 antibody) that demonstrated 84% nucleotide homology and 99% deduced amino acid homology with chicken brain calmodulin (CaM). This new CaM-like cDNA was named neoCaM. There is little nucleotide homology between the CaBP-28 cDNA and neoCaM. The CaBP-28 cDNA hybridizes with three transcripts of 2000, 2900, and 3300 bases which are dramatically induced by 1,25-dihydroxyvitamin D/sub 3/ (1,25(OH)/sub 2/D/sub 3/), while the neoCaM cDNA recognizes three distinct (from CaBP-28) transcripts. Two of these mRNAs are 1400 and 1800 bases as described for brain CaM, but another large 4000-base transcript is detected with neoCaM. Neither the CaM nor the neoCaM transcript reveals any modulation by 1,25(OH)/sub 2/D/sub 3/. Herein, the authors discuss the possible significance of not only the isolation of both cDNAs with a single antibody but also the relation of neoCaM to other well-characterized CaM cDNAs.

  5. Physiologically driven avian vocal synthesizer

    Science.gov (United States)

    Sitt, Jacobo D.; Arneodo, Ezequiel M.; Goller, Franz; Mindlin, Gabriel B.

    2010-03-01

    In this work, we build an electronic syrinx, i.e., a programmable electronic device capable of integrating biomechanical model equations for the avian vocal organ in order to synthesize song. This vocal prosthesis is controlled by the bird’s neural instructions to respiratory and the syringeal motor systems, thus opening great potential for studying motor control and its modification by sensory feedback mechanisms. Furthermore, a well-functioning subject-controlled vocal prosthesis can lay the foundation for similar devices in humans and thus provide directly health-related data and procedures.

  6. Novel procedure to compute a contact zone magnitude of vibrations of two-layered uncoupled plates

    Directory of Open Access Journals (Sweden)

    Awrejcewicz J.

    2005-01-01

    Full Text Available A novel iteration procedure for dynamical problems, where in each time step, a contacting plates' zone is improved, is proposed. Therefore, a zone and magnitude of a contact load are also improved. Investigations of boundary conditions' influence on externally driven vibrations of uncoupled two-layer plates, where for each of the layers, the Kirchhoff hypothesis holds, are carried out.

  7. Uncoupled achromatic condition of a dog-leg system with the presence of RF cavities

    CERN Document Server

    Geng, Huiping

    2013-01-01

    To merge the beam from either of the two injectors to the main linac, a dog-leg system will be employed in the second Medium Energy Beam Transport (MEBT2) line of the China ADS driving accelerator. The achromatic condition has to be guaranteed to avoid beam center excursion against energy jitter. RF cavities were found indispensable to control the bunch length growth in the dog-leg system of MEBT2. The full uncoupling between transverse and longitudinal plane is desired to minimize the growth of projected rms emittances. The uncoupled achromatic condition of this dogleg system with the presence of RF bunching cavities will be deduced using the method of transfer matrixes. It is found that to fulfil the uncoupling condition, the distance between the bunching cavities is uniquely determined by the maximum energy gain of the RF cavities. The theoretical analysis is verified by the simulation code TraceWin. The space charge effect on the uncoupled achromatic condition and the beam emittance growth will also be di...

  8. Single Cell Microgel Based Modular Bioinks for Uncoupled Cellular Micro- and Macroenvironments.

    Science.gov (United States)

    Kamperman, Tom; Henke, Sieger; van den Berg, Albert; Shin, Su Ryon; Tamayol, Ali; Khademhosseini, Ali; Karperien, Marcel; Leijten, Jeroen

    2017-02-01

    Modular bioinks based on single cell microgels within distinct injectable prepolymers enable uncoupling of biomaterials' micro- and macroenvironments. These inks allow biofabrication of 3D constructs that recapitulate the multiscale modular design of native tissues with a single cell resolution. This approach represents a major step forward in endowing engineered constructs with the multifunctionality that underlies the behavior of native tissues.

  9. Single Cell Microgel Based Modular Bioinks for Uncoupled Cellular Micro- and Macrenvironments

    NARCIS (Netherlands)

    Kamperman, T.; Henke, S.J.; Berg, van den A.; Shin, S.R.; Tamayol, A.; Khademhosseini, A.; Karperien, H.B.J.; Leijten, J.C.H.

    2016-01-01

    Modular bioinks based on single cell microgels within distinct injectable prepolymers enable uncoupling of biomaterials' micro- and macroenvironments. These inks allow biofabrication of 3D constructs that recapitulate the multiscale modular design of native tissues with a single cell resolution. Thi

  10. Emergence of a novel avian pox disease in British tit species.

    Directory of Open Access Journals (Sweden)

    Becki Lawson

    Full Text Available Avian pox is a viral disease with a wide host range. In Great Britain, avian pox in birds of the Paridae family was first diagnosed in a great tit (Parus major from south-east England in 2006. An increasing number of avian pox incidents in Paridae have been reported each year since, indicative of an emergent infection. Here, we utilise a database of opportunistic reports of garden bird mortality and morbidity to analyse spatial and temporal patterns of suspected avian pox throughout Great Britain, 2006-2010. Reports of affected Paridae (211 incidents outnumbered reports in non-Paridae (91 incidents. The majority (90% of Paridae incidents involved great tits. Paridae pox incidents were more likely to involve multiple individuals (77.3% than were incidents in non-Paridae hosts (31.9%. Unlike the small wart-like lesions usually seen in non-Paridae with avian pox in Great Britain, lesions in Paridae were frequently large, often with an ulcerated surface and caseous core. Spatial analyses revealed strong clustering of suspected avian pox incidents involving Paridae hosts, but only weak, inconsistent clustering of incidents involving non-Paridae hosts. There was no spatial association between Paridae and non-Paridae incidents. We documented significant spatial spread of Paridae pox from an origin in south-east England; no spatial spread was evident for non-Paridae pox. For both host clades, there was an annual peak of reports in August/September. Sequencing of the avian poxvirus 4b core protein produced an identical viral sequence from each of 20 great tits tested from Great Britain. This sequence was identical to that from great tits from central Europe and Scandinavia. In contrast, sequence variation was evident amongst virus tested from 17 non-Paridae hosts of 5 species. Our findings show Paridae pox to be an emerging infectious disease in wild birds in Great Britain, apparently originating from viral incursion from central Europe or Scandinavia.

  11. Crucial role of membrane potential in heat stress-induced overproduction of reactive oxygen species in avian skeletal muscle mitochondria.

    Science.gov (United States)

    Kikusato, Motoi; Toyomizu, Masaaki

    2013-01-01

    Heat stress is an environmental factor that causes oxidative stress. We found previously that acute heat stress stimulates the production of reactive oxygen species (ROS) in the skeletal muscle mitochondria of birds, and that this was accompanied by an increase of the mitochondrial membrane potential (ΔΨ) due to increased substrate oxidation by the electron transport chain. We also showed that avian uncoupling protein (avUCP) expression is decreased by the heat exposure. The present study clarifies whether ΔΨ is a major determinant of the overproduction of ROS due to acute heat stress, and if the decrease in avUCP expression is responsible for the elevation in ΔΨ. Control (24°C) and acute heat-stressed (34°C for 12 h) birds exhibited increased succinate-driven mitochondrial ROS production as indicated by an elevation of ΔΨ, with this increase being significantly higher in the heat-stressed group compared with the control group. In glutamate/malate-energized mitochondria, no difference in the ROS production between the groups was observed, though the mitochondrial ΔΨ was significantly higher in the heat-stressed groups compared with the control group. Furthermore, mitochondria energized with either succinate/glutamate or succinate/malate showed increased ROS production and ΔΨ in the heat-stressed group compared with mitochondria from the control group. These results suggest that succinate oxidation could play an important role in the heat stress-induced overproduction of mitochondrial ROS in skeletal muscle. In agreement with the notion of a decrease in avUCP expression in response to heat stress, proton leak, which was likely mediated by UCP (that part which is GDP-inhibited and arachidonic acid-sensitive), was reduced in the heat-exposed group. We suggest that the acute heat stress-induced overproduction of mitochondrial ROS may depend on ΔΨ, which may in turn result not only from increased substrate oxidation but also from a decrease in the

  12. Assessment of the cross-protective capability of recombinant capsid proteins derived from pig, rat, and avian hepatitis E viruses (HEV) against challenge with a genotype 3 HEV in pigs.

    Science.gov (United States)

    Sanford, Brenton J; Opriessnig, Tanja; Kenney, Scott P; Dryman, Barbara A; Córdoba, Laura; Meng, Xiang-Jin

    2012-09-28

    Hepatitis E virus (HEV), the causative agent of hepatitis E, is primarily transmitted via the fecal-oral route through contaminated water supplies, although many sporadic cases of hepatitis E are transmitted zoonotically via direct contact with infected animals or consumption of contaminated animal meats. Genotypes 3 and 4 HEV are zoonotic and infect humans and other animal species, whereas genotypes 1 and 2 HEV are restricted to humans. There exists a single serotype of HEV, although the cross-protective ability among the animal HEV strains is unknown. Thus, in this study we expressed and characterized N-terminal truncated ORF2 capsid antigens derived from swine, rat, and avian HEV strains and evaluated their cross-protective ability in a pig challenge model. Thirty, specific-pathogen-free, pigs were divided into 5 groups of 6 pigs each, and each group of pigs were vaccinated with 200 μg of swine HEV, rat HEV, or avian HEV ORF2 antigen or PBS buffer (2 groups) as positive and negative control groups. After a booster dose immunization at 2 weeks post-vaccination, the vaccinated animals all seroconverted to IgG anti-HEV. At 4 weeks post-vaccination, the animals were intravenously challenged with a genotype 3 mammalian HEV, and necropsied at 4 weeks post-challenge. Viremia, fecal virus shedding, and liver histological lesions were compared to assess the protective and cross-protective abilities of these antigens against HEV challenge in pigs. The results indicated that pigs vaccinated with truncated recombinant capsid antigens derived from three animal strains of HEV induced a strong IgG anti-HEV response in vaccinated pigs, but these antigens confer only partial cross-protection against a genotype 3 mammalian HEV. The results have important implications for the efficacy of current vaccines and for future vaccine development, especially against the novel zoonotic animal strains of HEV.

  13. Emerging and reemerging diseases of avian wildlife

    Science.gov (United States)

    Pello, Susan J.; Olsen, Glenn H.

    2013-01-01

    Of the many important avian wildlife diseases, aspergillosis, West Nile virus, avipoxvirus, Wellfleet Bay virus, avian influenza, and inclusion body disease of cranes are covered in this article. Wellfleet Bay virus, first identified in 2010, is considered an emerging disease. Avian influenza and West Nile virus have recently been in the public eye because of their zoonotic potential and links to wildlife. Several diseases labeled as reemerging are included because of recent outbreaks or, more importantly, recent research in areas such as genomics, which shed light on the mechanisms whereby these adaptable, persistent pathogens continue to spread and thrive.

  14. Worldwide phylogenetic relationship of avian poxviruses

    Science.gov (United States)

    Gyuranecz, Miklós; Foster, Jeffrey T.; Dán, Ádám; Ip, Hon S.; Egstad, Kristina F.; Parker, Patricia G.; Higashiguchi, Jenni M.; Skinner, Michael A.; Höfle, Ursula; Kreizinger, Zsuzsa; Dorrestein, Gerry M.; Solt, Szabolcs; Sós, Endre; Kim, Young Jun; Uhart, Marcela; Pereda, Ariel; González-Hein, Gisela; Hidalgo, Hector; Blanco, Juan-Manuel; Erdélyi, Károly

    2013-01-01

    Poxvirus infections have been found in 230 species of wild and domestic birds worldwide in both terrestrial and marine environments. This ubiquity raises the question of how infection has been transmitted and globally dispersed. We present a comprehensive global phylogeny of 111 novel poxvirus isolates in addition to all available sequences from GenBank. Phylogenetic analysis of Avipoxvirus genus has traditionally relied on one gene region (4b core protein). In this study we have expanded the analyses to include a second locus (DNA polymerase gene), allowing for a more robust phylogenetic framework, finer genetic resolution within specific groups and the detection of potential recombination. Our phylogenetic results reveal several major features of avipoxvirus evolution and ecology and propose an updated avipoxvirus taxonomy, including three novel subclades. The characterization of poxviruses from 57 species of birds in this study extends the current knowledge of their host range and provides the first evidence of the phylogenetic effect of genetic recombination of avipoxviruses. The repeated occurrence of avian family or order-specific grouping within certain clades (e.g. starling poxvirus, falcon poxvirus, raptor poxvirus, etc.) indicates a marked role of host adaptation, while the sharing of poxvirus species within prey-predator systems emphasizes the capacity for cross-species infection and limited host adaptation. Our study provides a broad and comprehensive phylogenetic analysis of the Avipoxvirus genus, an ecologically and environmentally important viral group, to formulate a genome sequencing strategy that will clarify avipoxvirus taxonomy.

  15. Replication-Uncoupled Histone Deposition during Adenovirus DNA Replication

    OpenAIRE

    Komatsu, Tetsuro; Nagata, Kyosuke

    2012-01-01

    In infected cells, the chromatin structure of the adenovirus genome DNA plays critical roles in its genome functions. Previously, we reported that in early phases of infection, incoming viral DNA is associated with both viral core protein VII and cellular histones. Here we show that in late phases of infection, newly synthesized viral DNA is also associated with histones. We also found that the knockdown of CAF-1, a histone chaperone that functions in the replication-coupled deposition of his...

  16. Role of nitric oxide synthase uncoupling at rostral ventrolateral medulla in redox-sensitive hypertension associated with metabolic syndrome.

    Science.gov (United States)

    Wu, Kay L H; Chao, Yung-Mei; Tsay, Shiow-Jen; Chen, Chen Hsiu; Chan, Samuel H H; Dovinova, Ima; Chan, Julie Y H

    2014-10-01

    Metabolic syndrome (MetS), which is rapidly becoming prevalent worldwide, is long known to be associated with hypertension and recently with oxidative stress. Of note is that oxidative stress in the rostral ventrolateral medulla (RVLM), where sympathetic premotor neurons reside, contributes to sympathoexcitation and hypertension. This study sought to identify the source of tissue oxidative stress in RVLM and their roles in neural mechanism of hypertension associated with MetS. Adult normotensive rats subjected to a high-fructose diet for 8 weeks developed metabolic traits of MetS, alongside increases in sympathetic vasomotor activity and blood pressure. In RVLM of these MetS rats, the tissue level of reactive oxygen species was increased, nitric oxide (NO) was decreased, and mitochondrial electron transport capacity was reduced. Whereas the protein expression of neuronal NO synthase (nNOS) or protein inhibitor of nNOS was increased, the ratio of nNOS dimer/monomer was significantly decreased. Oral intake of pioglitazone or intracisternal infusion of tempol or coenzyme Q10 significantly abrogated all those molecular events in high-fructose diet-fed rats and ameliorated sympathoexcitation and hypertension. Gene silencing of protein inhibitor of nNOS mRNA in RVLM using lentivirus carrying small hairpin RNA inhibited protein inhibitor of nNOS expression, increased the ratio of nNOS dimer/monomer, restored NO content, and alleviated oxidative stress in RVLM of high-fructose diet-fed rats, alongside significantly reduced sympathoexcitation and hypertension. These results suggest that redox-sensitive and protein inhibitor of nNOS-mediated nNOS uncoupling is engaged in a vicious cycle that sustains the production of reactive oxygen species in RVLM, resulting in sympathoexcitation and hypertension associated with MetS.

  17. Montana 2006 Avian Influenza Surveillance Project Report

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — During the summer of 2006, the U.S. Department of Agriculture (USDA) and the U.S. Fish and Wildlife Service (USFWS) initiated a nationwide avian influenza...

  18. Avian Habitat Data; Seward Peninsula, Alaska, 2012

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — This data product contains avian habitat data collected on the Seward Peninsula, Alaska, USA, during 21 May – 10 June 2012. We conducted replicated 10-min surveys at...

  19. Avian models in teratology and developmental toxicology.

    Science.gov (United States)

    Smith, Susan M; Flentke, George R; Garic, Ana

    2012-01-01

    The avian embryo is a long-standing model for developmental biology research. It also has proven utility for toxicology research both in ovo and in explant culture. Like mammals, avian embryos have an allantois and their developmental pathways are highly conserved with those of mammals, thus avian models have biomedical relevance. Fertile eggs are inexpensive and the embryo develops rapidly, allowing for high-throughput. The chick genome is sequenced and significant molecular resources are available for study, including the ability for genetic manipulation. The absence of a placenta permits the direct study of an agent's embryotoxic effects. Here, we present protocols for using avian embryos in toxicology research, including egg husbandry and hatch, toxicant delivery, and assessment of proliferation, apoptosis, and cardiac structure and function.

  20. Avian protection plan : Lostwood National Wildlife Refuge

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Lostwood National Wildlife Refuge (LNWR) initiated this Avian Protection Plan (APP) in 2003 to protect birds from potential electrocution hazards on the...

  1. Immunizing Canada geese against avian cholera

    Science.gov (United States)

    Price, J.I.

    1985-01-01

    A small flock of captive giant Canada geese were vaccinated with the experimental bac- terin in Nebraska to test its efficacy under field conditions. Only 2 of 157 vaccinates died from avian cholera during an annual spring die-off.

  2. Avian Influenza A Virus Infections in Humans

    Science.gov (United States)

    ... their saliva, mucous and feces. Human infections with bird flu viruses can happen when enough virus gets into ... Virus (CVV) for a Highly Pathogenic Avian Influenza (Bird Flu) Virus ” for more information on this process. ...

  3. Cell killing by avian leukosis viruses.

    OpenAIRE

    Weller, S K; Temin, H M

    1981-01-01

    Infection of chicken cells with a cytopathic avian leukosis virus resulted in the detachment of killed cells from the culture dish. The detached, dead cells contained more unintegrated viral DNA than the attached cells. These results confirm the hypothesis that cell killing after infection with a cytopathic avian leukosis virus is associated with accumulation of large amounts of unintegrated viral DNA. No accumulation of large amounts of integrated viral DNA was found in cells infected with c...

  4. Report of the Avian Development Working Group

    Science.gov (United States)

    Fallon, J. F.

    1985-01-01

    The anteroposterior axis of the avian embryo is established before it is laid. Baer's rule states that the cephalic end of the avian embryo will be away from the observer when the pointed end of the shell is on the observer's right. There are experimental data available which indicate gravity has a role in the establishment of the anteroposterior axis while the egg is in the uterus; this results in Baer's rule. The influence of gravity on egg development is studied.

  5. Unkempt is negatively regulated by mTOR and uncouples neuronal differentiation from growth control.

    Directory of Open Access Journals (Sweden)

    Amélie Avet-Rochex

    2014-09-01

    Full Text Available Neuronal differentiation is exquisitely controlled both spatially and temporally during nervous system development. Defects in the spatiotemporal control of neurogenesis cause incorrect formation of neural networks and lead to neurological disorders such as epilepsy and autism. The mTOR kinase integrates signals from mitogens, nutrients and energy levels to regulate growth, autophagy and metabolism. We previously identified the insulin receptor (InR/mTOR pathway as a critical regulator of the timing of neuronal differentiation in the Drosophila melanogaster eye. Subsequently, this pathway has been shown to play a conserved role in regulating neurogenesis in vertebrates. However, the factors that mediate the neurogenic role of this pathway are completely unknown. To identify downstream effectors of the InR/mTOR pathway we screened transcriptional targets of mTOR for neuronal differentiation phenotypes in photoreceptor neurons. We identified the conserved gene unkempt (unk, which encodes a zinc finger/RING domain containing protein, as a negative regulator of the timing of photoreceptor differentiation. Loss of unk phenocopies InR/mTOR pathway activation and unk acts downstream of this pathway to regulate neurogenesis. In contrast to InR/mTOR signalling, unk does not regulate growth. unk therefore uncouples the role of the InR/mTOR pathway in neurogenesis from its role in growth control. We also identified the gene headcase (hdc as a second downstream regulator of the InR/mTOR pathway controlling the timing of neurogenesis. Unk forms a complex with Hdc, and Hdc expression is regulated by unk and InR/mTOR signalling. Co-overexpression of unk and hdc completely suppresses the precocious neuronal differentiation phenotype caused by loss of Tsc1. Thus, Unk and Hdc are the first neurogenic components of the InR/mTOR pathway to be identified. Finally, we show that Unkempt-like is expressed in the developing mouse retina and in neural stem

  6. Ecology and conservation biology of avian malaria

    Science.gov (United States)

    LaPointe, Dennis A.; Atkinson, Carter T.; Samuel, Michael D.

    2012-01-01

    Avian malaria is a worldwide mosquito-borne disease caused by Plasmodium parasites. These parasites occur in many avian species but primarily affect passerine birds that have not evolved with the parasite. Host pathogenicity, fitness, and population impacts are poorly understood. In contrast to continental species, introduced avian malaria poses a substantial threat to naive birds on Hawaii, the Galapagos, and other archipelagoes. In Hawaii, transmission is maintained by susceptible native birds, competence and abundance of mosquitoes, and a disease reservoir of chronically infected native birds. Although vector habitat and avian communities determine the geographic distribution of disease, climate drives transmission patterns ranging from continuous high infection in warm lowland forests, seasonal infection in midelevation forests, and disease-free refugia in cool high-elevation forests. Global warming is expected to increase the occurrence, distribution, and intensity of avian malaria across this elevational gradient and threaten high-elevation refugia, which is the key to survival of many susceptible Hawaiian birds. Increased temperatures may have already increased global avian malaria prevalence and contributed to an emergence of disease in New Zealand.

  7. The Recruitment of Brown Adipose Tissue and Expression of Uncoupling Protein Gene in Brandt's Vole during Cold Exposure%布氏田鼠冷暴露中褐色脂肪组织的增补及解偶联蛋白基因表达

    Institute of Scientific and Technical Information of China (English)

    王煜; 黄晨西; 等

    2001-01-01

    布氏田鼠(Microtus brandti)随机分组暴露在冷环境[12L∶12D,(4±2)℃]中12?h,1 d,3 d,7 d,14 d,21 d和28 d;对照组生活在温暖环境下[12L∶12D,(25±2)℃]。与对照组相比,布氏田鼠的褐色脂肪组织(BAT)重量在冷暴露12?h~3?d时降低,7~21?d时则增加。7~28?d冷暴露组动物的BAT总蛋白和总DNA含量均比对照组明显提高。冷环境中的布氏田鼠解偶联蛋白(UCP)的mRNA随时间的延长而表达上调,在冷暴露21?d时达到高峰。结果表明,冷暴露能够诱导布氏田鼠BAT细胞增补和UCP基因表达,从而使适应性产热增加。%Brandt' s voles (Microtus brandti) were randomly divided into seven groups and exposed to cold temperature[12L∶12D,(4±2)℃] for 12 hours,1,3 ,7,14,21 and 28 days respectively;the control group was kept in warm place [12L∶12D,(25±2)℃]. Compared with the control,the weight of BAT and the total contents of DNA in BAT decreased during cold exposure from 12 hours to 3 days,but increased significantly from 7 to 28 days. Cold exposure also induced the increase of protein contents of BAT. In molecular level,the contents of UCP mRNA in BAT increased significantly and reached peak at 21 days. The results suggested that cold exposure could induce the recruitment of BAT cell and the expression of UCP gene,resulting in the increase of adaptive thermogenesis in Brandt' s voles.

  8. Uncoupling of reading and IQ over time: empirical evidence for a definition of dyslexia.

    Science.gov (United States)

    Ferrer, Emilio; Shaywitz, Bennett A; Holahan, John M; Marchione, Karen; Shaywitz, Sally E

    2010-01-01

    Developmental dyslexia is defined as an unexpected difficulty in reading in individuals who otherwise possess the intelligence and motivation considered necessary for fluent reading, and who also have had reasonable reading instruction. Identifying factors associated with normative and impaired reading development has implications for diagnosis, intervention, and prevention. We show that in typical readers, reading and IQ development are dynamically linked over time. Such mutual interrelationships are not perceptible in dyslexic readers, which suggests that reading and cognition develop more independently in these individuals. To our knowledge, these findings provide the first empirical demonstration of a coupling between cognition and reading in typical readers and a developmental uncoupling between cognition and reading in dyslexic readers. This uncoupling was the core concept of the initial description of dyslexia and remains the focus of the current definitional model of this learning disability.

  9. Effects of the uncoupling agents FCCP and CCCP on the saltatory movements of cytoplasmic organelles.

    Science.gov (United States)

    Hollenbeck, P J; Bray, D; Adams, R J

    1985-02-01

    Two potent uncoupling agents, carbonylcyanide-4-trifluoromethoxyphenylhydrazone (FCCP) and carbonylcyanide-3-chlorophenylhydrazone (CCCP) inhibit the movement of organelles in neurites of chick sensory neurones in culture. FCCP applied for 30 minutes at 10 microM reduces the number of moving organelles by 78% and a similar treatment with CCCP causes a reduction of 47%. At 100 microM either compound abolishes all directed movements both in neurites and in cultured 3T3 cells. These effects are probably not due to the discharge of proton gradients since 2,4-dinitrophenol (DNP), at concentrations shown to uncouple mitochondria by the discharge of the permeant cationic fluorescent probe rhodamine 123, fails to inhibit cytoplasmic movements. The inhibition of cytoplasmic movements by FCCP and CCCP is likely to be a consequence of their inhibitory action on a variety of enzymes, including dynein and myosin ATPases, through a reaction with sulfhydryl groups.

  10. Forming limits in the hole-flanging process by coupled and uncoupled damage models

    Science.gov (United States)

    Kacem, A.; Jégat, A.; Krichen, A.; Manach, P. Y.

    2013-12-01

    The aim of this work is to identify the limits of the hole-flanging process under different conditions. A 3D finite element model was developed to predict failure in hole-flanging process for sheet aluminium alloys. The Gurson-Tvergaard-Needleman (GTN) coupled damage model and the Bao-Wierzbicki (BW) uncoupled damage model were used. The parameters of both coupled and uncoupled models were identified by inverse analysis based on uniaxial tensile test. Experiments were conducted to analyse the types of failure that appear during the process. Numerical results were compared with experimental datas to check the validity of both models in predicting failure during the hole-flanging process. The comparative study showed that the GTN model predicts more accurately almost all types of failure while fracture occurrence can be only predicted by the BW model.

  11. In vivo and in vitro effects of the mitochondrial uncoupler FCCP on microtubules.

    OpenAIRE

    Maro, B.; Marty, M C; Bornens, M

    1982-01-01

    FCCP (carbonylcyanide-p-trifluoromethoxyphenylhydrazone), a potent uncoupler of oxidative phosphorylation, induces the complete disruption of cellular microtubules. A further analysis of this effect on BHK21 cells has shown that a decrease in the number of microtubules can be observed 15 min after adding FCCP and there is complete disruption after 60 min. Regrowth of microtubules was initiated 30 min after removal of FCCP, in marked contrast with the rapid reversion observed when microtubules...

  12. The Role of Avian leukosis virus Thansmembrane Protein in Membrane Fusion and Immunosuppression%禽白血病病毒跨膜蛋白在膜融合及免疫抑制中的作用

    Institute of Scientific and Technical Information of China (English)

    刘青; 成子强

    2008-01-01

    禽白血病病毒(Avian leukosis virus,ALV)入侵宿主细胞的关键是病毒囊膜与宿主细胞膜的融合,ALV在此过程中采用病毒-宿主细胞膜融合机制,这一机制的关键是病毒与细胞受体结合后跨膜蛋白一系列构象的变化.此外,ALV入侵宿主细胞后会引起严重的免疫抑制,继而引发肿瘤的产生.在对一些与ALV有相同感染机制的病毒的研究中发现,引起免疫抑制的关键区也是跨膜蛋白.因此进一步在分子水平上深入研究跨膜蛋白有助于正确认识病毒侵染的本质,做到更为有效的预防与治疗ALV感染.

  13. 禽戊型肝炎病毒中国分离株ORF3蛋白的真核表达与抗原性分析%Eukaryotic Expression and Antigencity Analysis of Avian Hepatitis E Virus ORF3 Protein from China Isolate

    Institute of Scientific and Technical Information of China (English)

    胡守彬; 赵钦; 赵菲菲; 肖一红; 周恩民

    2012-01-01

    [目的]为获得禽戊型肝炎病毒(hepatitis E virus,HEV)中国分离株(CaHEV) ORF3基因重组蛋白,并对其抗原性进行分析.[方法]提取CaHEV总RNA,通过RT-PCR获得ORF3基因,构建重组质粒pFastBac-HTB-ORF3,转座DH10Bac感受态细胞,提取重组杆粒Bacmid-ORF3,转染sf9细胞,用SDS-PAGE、IFA和Western blot对重组蛋白表达进行鉴定.优化表达条件,将纯化的ORF3蛋白免疫小鼠制备多克隆抗体,用制备的多抗分别与截短表达的3段CaHEV ORF3蛋白进行Western blot和ELISA检测,分析ORF3蛋白的抗原表位区.[结果]SDS-PAGE、Western blot和IFA结果表明CaHEV ORF3蛋白被成功表达且存在细胞内,昆虫细胞在接毒后4天表达量最高.将ORF3重组蛋白免疫小鼠,利用间接ELISA方法,对制备的多抗倍比稀释检测,效价达到104,Western blot和ELISA分析表明ORF3蛋白C端74-88氨基酸处具有较强的抗原性.[结论]本试验用Bac-to-bac系统成功构建含有CaHEV ORF3基因的重组杆状病毒,并在昆虫细胞中得到表达,其主要抗原表位位于C端74-88氨基酸,为进一步研究禽HEV ORF3的蛋白结构和功能奠定了基础.%[Objective] The objective of the study is to obtain recombinant protein of avian hepatitis E virus (HEV) ORF3 of China isolate (CaHEV) and analyze its antigencity. [Method] Total RNA was extracted from CaHEV and ORF3 gene was amplified by RT-PCR and cloned into pFastBac-HT. The recombinant plasmid was transformed into DHlOBac and transfected into sf9 insect cells to express the recombinant ORF3 protein. The recombinant ORF3 protein was identified by SDS-PAGE, Western bolt and IFA methods. The purified ORF3 protein was used to immunize Balb/c mice to produce polyclonal antibodies and then to analyze the epitopes in ORF3 using 3 truncated ORF3 proteins [Result] The results showed that ORF3 protein was expressed with the highest expression level at 4 days post inoculation. The titer of anti-ORF3 polyclonal antibodies was 104

  14. Using avian radar to examine relationships among avian activity, bird strikes, and meteorological factors

    Science.gov (United States)

    Coates, Peter S.; Casazza, Michael L.; Halstead, Brian J.; Fleskes, Joseph P.; Laughlin, James A.

    2011-01-01

    Radar systems designed to detect avian activity at airfields are useful in understanding factors that influence the risk of bird and aircraft collisions (bird strikes). We used an avian radar system to measure avian activity at Beale Air Force Base, California, USA, during 2008 and 2009. We conducted a 2-part analysis to examine relationships among avian activity, bird strikes, and meteorological and time-dependent factors. We found that avian activity around the airfield was greater at times when bird strikes occurred than on average using a permutation resampling technique. Second, we developed generalized linear mixed models of an avian activity index (AAI). Variation in AAI was first explained by seasons that were based on average migration dates of birds at the study area. We then modeled AAI by those seasons to further explain variation by meteorological factors and daily light levels within a 24-hour period. In general, avian activity increased with decreased temperature, wind, visibility, precipitation, and increased humidity and cloud cover. These effects differed by season. For example, during the spring bird migration period, most avian activity occurred before sunrise at twilight hours on clear days with low winds, whereas during fall migration, substantial activity occurred after sunrise, and birds generally were more active at lower temperatures. We report parameter estimates (i.e., constants and coefficients) averaged across models and a relatively simple calculation for safety officers and wildlife managers to predict AAI and the relative risk of bird strike based on time, date, and meteorological values. We validated model predictability and assessed model fit. These analyses will be useful for general inference of avian activity and risk assessment efforts. Further investigation and ongoing data collection will refine these inference models and improve our understanding of factors that influence avian activity, which is necessary to inform

  15. Avian cholera in Nebraska's Rainwater Basin

    Science.gov (United States)

    Windingstad, R.M.; Hurt, J.J.; Trout, A.K.; Cary, J.

    1984-01-01

    The first report of avian cholera in North America occurred in northwestern Texas in winter 1944 (Quortrup et al. 1946). In 1975, mortality from avian cholera occurred for the first time in waterfowl in the Rainwater Basin of Nebraska when an estimated 25,000 birds died (Zinkl et al. 1977). Avian cholera has continued to cause mortality in wild birds in specific areas of the Basin each spring since. Losses of waterfowl from avian cholera continue to be much greater in some of the wetlands in the western part of the Basin than in the east. Several wetlands in the west have consistently higher mortality and are most often the wetlands where initial mortality is noticed each spring (Figure 1). The establishment of this disease in Nebraska is of considerable concern because of the importance of the Rainwater Basin as a spring staging area for waterfowl migrating to their breeding grounds. The wetlands in this area are on a major migration route used by an estimated 5 to 9 million ducks and several hundred thousand geese. A large portion of the western mid-continental greater white-fronted goose (Anser albifrons) population stage in the Basin each spring. Occasionally, whooping cranes (Grus americana) use these wetlands during migration, and lesser sandhill cranes (Grus canadensis) staging on the nearby Platte River sometimes use wetlands where avian cholera occurs (Anonymous 1981). Our objectives were to determine whether certain water quality variables in the Rainwater Basin differed between areas of high and low avian cholera incidence. These results would then be used for laboratory studies involving the survivability of Pasteurella multocida, the causative bacterium of avian cholera. Those studies will be reported elsewhere.

  16. Protein

    Science.gov (United States)

    ... Food Service Resources Additional Resources About FAQ Contact Protein Protein is found throughout the body—in muscle, ... the heart and respiratory system, and death. All Protein Isn’t Alike Protein is built from building ...

  17. Avian Point Count Locations - Dahomey NWR 2007-2008

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Map depicts locations of avian point counts conducted on Dahomey in 2007 and 2008. Actual point count data are contained in the avian knowledge network database

  18. Ras transformation uncouples the kinesin-coordinated cellular nutrient response

    Science.gov (United States)

    Zaganjor, Elma; Weil, Lauren M.; Gonzales, Joshua X.; Minna, John D.; Cobb, Melanie H.

    2014-01-01

    The kinesin family members (KIFs) KIF2A and KIF2C depolymerize microtubules, unlike the majority of other kinesins, which transport cargo along microtubules. KIF2A regulates the localization of lysosomes in the cytoplasm, which assists in activation of the mechanistic target of rapamycin complex 1 (mTORC1) on the lysosomal surface. We find that the closely related kinesin KIF2C also influences lysosomal organization in immortalized human bronchial epithelial cells (HBECs). Expression of KIF2C and, to a lesser extent, KIF2A in untransformed and mutant K-Ras–transformed cells is regulated by ERK1/2. Prolonged inhibition of ERK1/2 activation with PD0325901 mimics nutrient deprivation by disrupting lysosome organization and decreasing mTORC1 activity in HBEC, suggesting a long-term mechanism for optimization of mTORC1 activity by ERK1/2. We tested the hypothesis that up-regulation of KIF2C and KIF2A by ERK1/2 caused aberrant lysosomal positioning and mTORC1 activity in a mutant K-Ras–dependent cancer and cancer model. In Ras-transformed cells, however, mTORC1 activity and lysosome organization appear independent of ERK1/2 and these kinesins although ERK1/2 activity and the kinesins are required for Ras-dependent proliferation and migration. We conclude that mutant K-Ras repurposes these signaling and regulatory proteins to support the transformed phenotype. PMID:25002494

  19. Changes in the survival curve shape of E. coli cells following irradiation in the presence of uncouplers of oxidative phosphorylation.

    Science.gov (United States)

    Anderson, R F; Patel, K B; Evans, M D

    1985-10-01

    Four uncouplers of oxidative phosphorylation (UOP) (carbonyl cyanide m-chlorophenylhydrazone, 2,4-dinitrophenol, 4-hydroxybenzylidenemalonitrile and N-phenylanthranilic acid) have been found to alter the shape of the radiation survival curves of several cell lines of E. coli when present during irradiation in oxia. Incubation of cells with high concentrations of UOP for 30 min before irradiation induced an increase in extrapolation number (n) in cell lines AB 1157 (wild-type), AB 1886(uvrA-) and KMBL(polA-) but not GR 501(lig-)ts, AB 2463(recA-) and AB 2480(uvrA-recA-). In addition the UOP all effect a decrease in mean lethal dose (D0) even when tested at low concentrations or short contact times. Studies with wild-type cells correlate the increase in n with measured increased levels of ATP (above oxic control cells) produced upon incubation with UOP. The increased levels of ATP most likely arise from the UOP overstimulating glycolysis. The decrease in D0 cannot be associated with any of the repair pathways investigated and it is concluded that the highly lipophilic UOP directly or indirectly potentiate other target(s) to radiation damage as well as DNA under oxic conditions. Treatment of the cells with UOP did not result in the deleterious depletion of energy substrates, loss of non-protein thiols or the production of cytotoxins upon irradiation.

  20. Determination and analysis of the complete genomic sequence of avian hepatitis E virus (avian HEV) and attempts to infect rhesus monkeys with avian HEV.

    Science.gov (United States)

    Huang, F F; Sun, Z F; Emerson, S U; Purcell, R H; Shivaprasad, H L; Pierson, F W; Toth, T E; Meng, X J

    2004-06-01

    Avian hepatitis E virus (avian HEV), recently identified from a chicken with hepatitis-splenomegaly syndrome in the United States, is genetically and antigenically related to human and swine HEVs. In this study, sequencing of the genome was completed and an attempt was made to infect rhesus monkeys with avian HEV. The full-length genome of avian HEV, excluding the poly(A) tail, is 6654 bp in length, which is about 600 bp shorter than that of human and swine HEVs. Similar to human and swine HEV genomes, the avian HEV genome consists of a short 5' non-coding region (NCR) followed by three partially overlapping open reading frames (ORFs) and a 3'NCR. Avian HEV shares about 50 % nucleotide sequence identity over the complete genome, 48-51 % identity in ORF1, 46-48 % identity in ORF2 and only 29-34 % identity in ORF3 with human and swine HEV strains. Significant genetic variations such as deletions and insertions, particularly in ORF1 of avian HEV, were observed. However, motifs in the putative functional domains of ORF1, such as the helicase and methyltransferase, were relatively conserved between avian HEV and mammalian HEVs, supporting the conclusion that avian HEV is a member of the genus Hepevirus. Phylogenetic analysis revealed that avian HEV represents a branch distinct from human and swine HEVs. Swine HEV infects non-human primates and possibly humans and thus may be zoonotic. An attempt was made to determine whether avian HEV also infects across species by experimentally inoculating two rhesus monkeys with avian HEV. Evidence of virus infection was not observed in the inoculated monkeys as there was no seroconversion, viraemia, faecal virus shedding or serum liver enzyme elevation. The results from this study confirmed that avian HEV is related to, but distinct from, human and swine HEVs; however, unlike swine HEV, avian HEV is probably not transmissible to non-human primates.

  1. Mimicking a SURF1 allele reveals uncoupling of cytochrome c oxidase assembly from translational regulation in yeast.

    Science.gov (United States)

    Reinhold, Robert; Bareth, Bettina; Balleininger, Martina; Wissel, Mirjam; Rehling, Peter; Mick, David U

    2011-06-15

    Defects in mitochondrial energy metabolism lead to severe human disorders, mainly affecting tissues especially dependent on oxidative phosphorylation, such as muscle and brain. Leigh Syndrome describes a severe encephalomyopathy in infancy, frequently caused by mutations in SURF1. SURF1, termed Shy1 in Saccharomyces cerevisiae, is a conserved assembly factor for the terminal enzyme of the respiratory chain, cytochrome c oxidase. Although the molecular function of SURF1/Shy1 is still enigmatic, loss of function leads to cytochrome c oxidase deficiency and reduced expression of the central subunit Cox1 in yeast. Here, we provide insights into the molecular mechanisms leading to disease through missense mutations in codons of the most conserved amino acids in SURF1. Mutations affecting G(124) do not compromise import of the SURF1 precursor protein but lead to fast turnover of the mature protein within the mitochondria. Interestingly, an Y(274)D exchange neither affects stability nor localization of the protein. Instead, SURF1(Y274D) accumulates in a 200 kDa cytochrome c oxidase assembly intermediate. Using yeast as a model, we demonstrate that the corresponding Shy1(Y344D) is able to overcome the stage where cytochrome c oxidase assembly links to the feedback regulation of mitochondrial Cox1 expression. However, Shy1(Y344D) impairs the assembly at later steps, most apparent at low temperature and exhibits a dominant-negative phenotype upon overexpression. Thus, exchanging the conserved tyrosine (Y(344)) with aspartate in yeast uncouples translational regulation of Cox1 from cytochrome c oxidase assembly and provides evidence for the dual functionality of Shy1.

  2. Proceedings of National Avian-Wind Power Planning Meeting IV

    Energy Technology Data Exchange (ETDEWEB)

    NWCC Avian Subcommittee

    2001-05-01

    OAK-B135 The purpose of the fourth meeting was to (1) share research and update research conducted on avian wind interactions (2) identify questions and issues related to the research results, (3) develop conclusions about some avian/wind power issues, and (4) identify questions and issues for future avian research.

  3. 9 CFR 113.325 - Avian Encephalomyelitis Vaccine.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Avian Encephalomyelitis Vaccine. 113... REQUIREMENTS Live Virus Vaccines § 113.325 Avian Encephalomyelitis Vaccine. Avian Encephalomyelitis Vaccine... vaccine production. All serials shall be prepared from the first through the fifth passage from the...

  4. 解耦联蛋白研究进展%The Study of Uncoupling Protein

    Institute of Scientific and Technical Information of China (English)

    屈金亭

    2008-01-01

    解耦联蛋白(UCP)是线粒体内膜上重要的转运蛋白质,具有解偶联活性,限制ATP合成,增加产热,参与糖及脂肪酸的利用,与能量平衡、代谢、体重调节关系密切.

  5. Effects of central irisin administration on the uncoupling proteins in rat brain.

    Science.gov (United States)

    Erden, Yavuz; Tekin, Suat; Sandal, Suleyman; Onalan, Ebru Etem; Tektemur, Ahmet; Kirbag, Sevda

    2016-04-01

    Irisin is a thermogenic peptide that enables the development of brown adipose tissue from white adipose tissue by activating the UCP1. This study has been designed to determine the effects of the irisin on UCPs. Sprague Dawley female rats were used in the study. 1, 3 and 10μM concentrations of irisin were injected intracerebroventricularly to the rats, and the control group was received only vehicle. The animals were killed at the 16, 24, and 48h time intervals and their brains were taken out. The hypothalamus, pituitary gland, hippocampus, cerebellum, striatum and cortex areas were separated and the UCP2, UCP3, UCP4 and UCP5 mRNA levels were determined. Just before the animals were killed, their body temperatures were recorded. It was observed that after application of the high dose irisin, UCP5 mRNA level in the all brain areas increased (pbrain areas (except the pituitary gland; pbrain, and the irisin affects this expression, and may have effective roles in some brain functions.

  6. Effects of Mitochondrial Uncoupling Protein 2 Inhibition by Genipin in Human Cumulus Cells

    Directory of Open Access Journals (Sweden)

    Hongshan Ge

    2015-01-01

    Full Text Available UCP2 plays a physiological role by regulating mitochondrial biogenesis, maintaining energy balance, ROS elimination, and regulating cellular autophagy in numerous tissues. But the exact roles of UCP2 in cumulus cells are still not clear. Genipin, a special UCP2 inhibitor, was added into the cultural medium to explore the roles of UCP2 in human cumulus cells. There were no significant differences in ATP and mitochondrial membrane potential levels in cumulus cells from UCP2 inhibiting groups as compared with the control. The levels of ROS and Mn-SOD were markedly elevated after UCP2 inhibited Genipin. However, the ratio of reduced GSH to GSSG significantly declined after treatment with Genipin. UCP2 inhibition by Genipin also resulted in obvious increase in the active caspase-3, which accompanied the decline of caspase-3 mRNA. The level of progesterone in culture medium declined obviously after Genipin treatment. But there was no significant difference in estradiol concentrations. This study indicated that UCP2 is expressed in human cumulus cells and plays important roles on mediate ROS production, apoptotic process, and steroidogenesis, suggesting UCP2 may be involved in regulation of follicle development and oocyte maturation and quality.

  7. Avian cytochrome P450 (CYP 1-3 family genes: isoforms, evolutionary relationships, and mRNA expression in chicken liver.

    Directory of Open Access Journals (Sweden)

    Kensuke P Watanabe

    Full Text Available Cytochrome P450 (CYP of chicken and other avian species have been studied primarily with microsomes or characterized by cloning and protein expression. However, the overall existing isoforms in avian CYP1-3 families or dominant isoforms in avian xenobiotic metabolism have not yet been elucidated. In this study, we aimed to clarify and classify all of the existing isoforms of CYP1-3 in avian species using available genome assemblies for chicken, zebra finch, and turkey. Furthermore, we performed qRT-PCR assay to identify dominant CYP genes in chicken liver. Our results suggested that avian xenobiotic-metabolizing CYP genes have undergone unique evolution such as CYP2C and CYP3A genes, which have undergone avian-specific gene duplications. qRT-PCR experiments showed that CYP2C45 was the most highly expressed isoform in chicken liver, while CYP2C23b was the most highly induced gene by phenobarbital. Considering together with the result of further enzymatic characterization, CYP2C45 may have a dominant role in chicken xenobiotic metabolism due to the constitutive high expression levels, while CYP2C23a and CYP2C23b can be greatly induced by chicken xenobiotic receptor (CXR activators. These findings will provide not only novel insights into avian xenobiotic metabolism, but also a basis for the further characterization of each CYP gene.

  8. Assembly and immunological properties of a bivalent virus-like particle (VLP) for avian influenza and Newcastle disease.

    Science.gov (United States)

    Shen, Huifang; Xue, Chunyi; Lv, Lishan; Wang, Wei; Liu, Qiliang; Liu, Kang; Chen, Xianxian; Zheng, Jing; Li, Xiaoming; Cao, Yongchang

    2013-12-26

    Avian influenza virus (AIV) and Newcastle disease virus (NDV) are both important pathogens in poultry worldwide. The protection of poultry from avian influenza and Newcastle disease can be achieved through vaccination. We embarked on the development of a bivalent vaccine that would allow for a single immunization against both avian influenza and Newcastle disease. We constructed a chimeric virus-like particle (VLP) that is composed of the M1 protein and HA protein of avian influenza virus and a chimeric protein containing the cytoplasmic and transmembrane domains of AIV neuraminidase protein (NA) and the ectodomain of the NDV hemagglutinin-neuraminidase (HN) protein (NA/HN). The single immunization of chickens with the chimeric VLP vaccine induced both AIV H5- and NDV-specific antibodies. The HI titers and specific antibodies elicited by the chimeric VLPs were statistically similar to those elicited in animals vaccinated with the corresponding commercial monovalent vaccines. Chickens vaccinated with chimeric VLP vaccine and then challenged with the Newcastle disease F48E9 virus displayed complete protection. Overall, the chimeric VLP vaccine elicits strong immunity and can protect against Newcastle disease virus challenge.

  9. Cloning and Expression of Highly Pathogenic Avian Influenza Virus Full-Length Nonstructural Gene in Pichia pastoris

    Directory of Open Access Journals (Sweden)

    M. B. Abubakar

    2011-01-01

    Full Text Available Avian influenza (AI is a highly contagious and rapidly evolving pathogen of major concern to the poultry industry and human health. Rapid and accurate detection of avian influenza virus is a necessary tool for control of outbreaks and surveillance. The AI virus A/Chicken/Malaysia/5858/2004 (H5N1 was used as a template to produce DNA clones of the full-length NS1 genes via reverse transcriptase synthesis of cDNA by PCR amplification of the NS1 region. Products were cloned into pCR2.0 TOPO TA plasmid and subsequently subcloned into pPICZαA vector to construct a recombinant plasmid. Recombinant plasmid designated as pPICZαA-NS1 gene was confirmed by PCR colony screening, restriction enzyme digestion, and nucleotide sequence analysis. The recombinant plasmid was transformed into Pichia pastoris GS115 strain by electroporation, and expressed protein was identified by SDS-PAGE and western blotting. A recombinant protein of approximately ~28 kDa was produced. The expressed protein was able to bind a rabbit polyclonal antibody of nonstructural protein (NS1 avian influenza virus H5N1. The result of the western blotting and solid-phase ELISA assay using H5N1 antibody indicated that the recombinant protein produced retained its antigenicity. This further indicates that Pichia pastoris could be an efficient expression system for a avian influenza virus nonstructural (NS1.

  10. Subtype Identification of Avian Influenza Virus on DNA Microarray

    Institute of Scientific and Technical Information of China (English)

    WANG Xiu-rong; YU Kang-zhen; DENG Guo-hua; SHI Rui; LIU Li-ling; QIAO Chuan-ling; BAO Hong-mei; KONG Xian-gang; CHEN Hua-lan

    2005-01-01

    We have developed a rapid microarray-based assay for the reliable detection of H5, H7 and H9 subtypes of avian influenza virus (AIV). The strains used in the experiment were A/Goose/Guangdong/1/96 (H5N1), A/African starling/983/79 (H7N1) and A/Turkey/Wiscosin/1/66 (H9N2). The capture DNAs clones which encoding approximate 500-bp avian influenza virus gene fragments obtained by RT-PCR, were spotted on a slide-bound microarray. Cy5-1abeled fluorescent cDNAs,which generated from virus RNA during reverse transcription were hybridized to these capture DNAs. These capture DNAs contained multiple fragments of the hemagglutinin and matrix protein genes of AIV respectively, for subtyping and typing AIV. The arrays were scanned to determine the probe binding sites. The hybridization pattern agreed approximately with the known grid location of each target. The results show that DNA microarray technology provides a useful diagnostic method for AIV.

  11. THE AVIAN FLU IMPACT ON THE ROMANIAN POULTRY MARKET

    Directory of Open Access Journals (Sweden)

    Silvius Stanciu

    2015-05-01

    Full Text Available Poultry meat represents one of the most dynamic branches of the local meat production. The poultry sector represents a good quality protein source, at an acceptable price as compared to other animal production domains. There has been an ascending evolution of the sector after the year 2000, although there appeared a series of discontinuities that affected agricultural production, mainly on a short-term basis. The Avian Flu led to 190 million euros’ worth losses at the level of Romanian national economy. Low consumption due to the impact was a short-term consequence, being rapidly amortized by the Romanian producers. The lack of some business continuity insurance measures can further affect the poultry meat sector, which does not have the necessary robustness needed in case of larger shocks. The following article proposes an analysis of the Avian Flu crisis economic effects on the Romanian meat sector, and it is part of a general framework of research regarding the Romanian food chain resilience to critical situations.

  12. Thermal emissivity of avian eggshells.

    Science.gov (United States)

    Björn, Lars Olof; Bengtson, Sven-Axel; Li, Shaoshan; Hecker, Christoph; Ullah, Saleem; Roos, Arne; Nilsson, Annica M

    2016-04-01

    The hypothesis has been tested that evolution has resulted in lower thermal emissivity of eggs of birds breeding openly in cold climates than of eggs of birds that nest under protective covering or in warmer climates. Directional thermal emissivity has been estimated from directional-hemispherical reflectance spectra. Due to several methodological difficulties the absolute emissivity is not accurately determined, but differences between species are obvious. Most notably, small waders of the genus Calidris, breeding in cold climates on the tundra, and in most cases with uniparental nest attendance, have low directional emissivity of their eggshells, about 0.92 when integration is carried out for wavelengths up to 16μm. Species belonging to Galloanserinae have the highest directional emissivity, about 0.96, of their eggs. No differences due to climate or breeding conditions were found within this group. Eggs of most other birds tested possess intermediate emissivity, but the values for Pica pica and Corvus corone cornix are as low as for Calidris. Large species-dependent differences in spectral reflectance were found at specific wavelengths. For instance, at 4.259μm the directional-hemispherical reflectance for galliforms range from 0.05 to 0.09, while for Fratercula arctica and Fulmarus glacialis it is about 0.3. The reflection peaks at 6.5 and 11.3μm due to calcite are differentially attenuated in different species. In conclusion, the hypothesis that evolution has resulted in lower thermal emissivity of bird eggs being exposed in cold climates is not supported by our results. The emissivity is not clearly related to nesting habits or climate, and it is unlikely that the small differences observed are ecologically important. The spectral differences between eggs that nevertheless exist should be taken into account when using infrared thermometers for estimating the surface temperature of avian eggs.

  13. Economic effects of avian influenza on egg producers in Turkey

    Directory of Open Access Journals (Sweden)

    V Demircan

    2009-09-01

    Full Text Available This study determined the economic effects of avian influenza on the egg-production sector of Afyon Province, Turkey. Economic indicators were compared before and during the avian influenza outbreak. A questionnaire was conducted with 75 poultry farmers. Farms were divided into three groups according to their size. The profitability of the three farm size groups was compared during two study periods: before and during the avian influenza outbreak. The results indicate that, as compared to previous levels, farms experienced significantly reduced incomes during the avian influenza episode. While net income and profit margin were found to be negative in all three farm groups during the avian influenza period, only group I showed economic loss prior to avian influenza. Average net income per group was -19,576.14, -39,810.11, and -112,035.33 YTL respectively during the avian influenza outbreak, compared with prior incomes of -5,665.51, 8,422.92, and 16,3873.71 YTL (1 USD=1.43 YTL. The profit margin per egg during avian influenza was -0.029, -0.016, -0.010 YTL in group I, II, III, respectively, as compared to -0.007, 0.003, and 0.014 YTL/egg before avian influenza. It was found that, whereas larger farms were more profitable than small farms prior to the avian influenza period, larger farms suffered greater economic losses than small farms during avian influenza outbreak in the participating farms.

  14. Suppression of injuries caused by a lytic RNA virus (mengovirus) and their uncoupling from viral reproduction by mutual cell/virus disarmament.

    Science.gov (United States)

    Mikitas, Olga V; Ivin, Yuri Y; Golyshev, Sergey A; Povarova, Natalia V; Galkina, Svetlana I; Pletjushkina, Olga Y; Nadezhdina, Elena S; Gmyl, Anatoly P; Agol, Vadim I

    2012-05-01

    Viruses often elicit cell injury (cytopathic effect [CPE]), a major cause of viral diseases. CPE is usually considered to be a prerequisite for and/or consequence of efficient viral growth. Recently, we proposed that viral CPE may largely be due to host defensive and viral antidefensive activities. This study aimed to check the validity of this proposal by using as a model HeLa cells infected with mengovirus (MV). As we showed previously, infection of these cells with wild-type MV resulted in necrosis, whereas a mutant with incapacitated antidefensive ("security") viral leader (L) protein induced apoptosis. Here, we showed that several major morphological and biochemical signs of CPE (e.g., alterations in cellular and nuclear shape, plasma membrane, cytoskeleton, chromatin, and metabolic activity) in cells infected with L(-) mutants in the presence of an apoptosis inhibitor were strongly suppressed or delayed for long after completion of viral reproduction. These facts demonstrate that the efficient reproduction of a lytic virus may not directly require development of at least some pathological alterations normally accompanying infection. They also imply that L protein is involved in the control of many apparently unrelated functions. The results also suggest that the virus-activated program with competing necrotic and apoptotic branches is host encoded, with the choice between apoptosis and necrosis depending on a variety of intrinsic and extrinsic conditions. Implementation of this defensive suicidal program could be uncoupled from the viral reproduction. The possibility of such uncoupling has significant implications for the pathogenesis and treatment of viral diseases.

  15. Synchronization of uncoupled excitable systems induced by white and coloured noise

    Energy Technology Data Exchange (ETDEWEB)

    Zambrano, Samuel; Marino, Ines P; Seoane, Jesus M; Sanjuan, Miguel A F [Nonlinear Dynamics, Chaos and Complex Systems Group, Departamento de Fisica, Universidad Rey Juan Carlos, Tulipan s/n, 28933 Mostoles, Madrid (Spain); Euzzor, Stefano; Geltrude, Andrea; Meucci, Riccardo; Arecchi, Fortunato T, E-mail: samuel.zambrano@urjc.e, E-mail: jesus.seoane@urjc.e, E-mail: ines.perez@urjc.e [CNR-Istituto Nazionale di Ottica Applicata, Largo E Fermi, 6 50125 Firenze (Italy)

    2010-05-15

    We study, both numerically and experimentally, the synchronization of uncoupled excitable systems due to a common noise. We consider two identical FitzHugh-Nagumo systems, which display both spiking and non-spiking behaviours in chaotic or periodic regimes. An electronic circuit provides a laboratory implementation of these dynamics. Synchronization is tested with both white and coloured noise, showing that coloured noise is more effective in inducing synchronization of the systems. We also study the effects on the synchronization of parameter mismatch and of the presence of intrinsic (not common) noise, and we conclude that the best performance of coloured noise is robust under these distortions.

  16. Evaluation of the electron transport chain inhibition and uncoupling of mitochondrial bioelectrocatalysis with antibiotics and nitro-based compounds

    Energy Technology Data Exchange (ETDEWEB)

    Arechederra, Marguerite N.; Fischer, Caitlin N.; Wetzel, David J. [Department of Chemistry, Saint Louis University, 3501 Laclede Ave., St. Louis, MO (United States); Minteer, Shelley D., E-mail: minteers@slu.ed [Department of Chemistry, Saint Louis University, 3501 Laclede Ave., St. Louis, MO (United States)

    2010-12-30

    Mitochondrial bioelectrocatalysis can be useful for sensing applications due to the unique metabolic pathways than can be selectively inhibited and uncoupled in mitochondria. This paper details the comparison of different inhibitors and nitro-containing explosive uncouplers in a mitochondria-catalyzed biofuel cell for self-powered explosive sensing. Previous research has reported inhibition of pyruvate oxidation at a mitochondria-modified electrode followed by nitroaromatic uncoupling of current and power. We have previously used oligomycin as the antibiotic and nitrobenzene as the uncoupler of the membrane in the mitochondria-catalyzed biofuel cell, but no comprehensive comparison of various mitochondria inhibitors or explosives has been performed. Results are discussed here for inhibitors targeting complex I, complex III, ATP synthases, adenine nucleotide transport and monocarboxylic acid transport. Reactivation with nitrobenzene was possible in the presence of these inhibitors: oligomycin, 3,3'-diindolylmethane, atractyloside, rotenone, {alpha}-cyano-4-hydroxy cinnamic acid and antimycin A. All eleven explosives studied, including: 2,4,6-trinitrotoluene (TNT) and 1,3,5-trinitroperhydro-1,3,5-triazine (RDX), caused uncoupling of the mitochondria function and could be detected by the biosensor.

  17. Tissue distribution of HSPA9/mortalin in avian species and its regulation by gender, genotype and heat stress

    Science.gov (United States)

    Heat shock 70kDa protein 9 (HSPA9)/mortalin is a multipotent chaperone regulating cellular processes ranging from stress response to energy homeostasis. HSPA9 has been extensively studied in mammals however there is a paucity of information in avian species. The present study aimed to characterize H...

  18. A reverse transcription-polymerase chain reaction assay for the detection of avian pneumovirus (Colorado strain).

    Science.gov (United States)

    Ali, A; Reynolds, D L

    1999-01-01

    A reverse transcription-polymerase chain reaction assay was developed for the detection of avian pneumovirus (Colorado strain) (APV-Col). The specific primers were designed from the published sequence of the matrix protein gene of APV-Col. The primers amplified a product of 631 nucleotides from APV-Col. The assay identified only APV-Col and did not react with Newcastle disease virus and infectious bronchitis virus.

  19. Integration and Validation of Avian Radars (IVAR)

    Science.gov (United States)

    2011-08-01

    operations; 1-year visual census; multiple eBirdRad radars; fiber- optic wired LAN (planned) NAS Patuxent River, MD X B X Medium-sized air station...introduced a multibeam avian radar antenna that purports to double the beam width (from 4° to 8°), while at the same time increasing the precision of the

  20. Avian influenza virus and Newcastle disease virus

    Science.gov (United States)

    Avian influenza virus (AIV) and Newcastle disease virus (NDV) severely impact poultry egg production. Decreased egg yield and hatchability, as well as misshapen eggs, are often observed during infection with AIV and NDV, even with low-virulence strains or in vaccinated flocks. Data suggest that in...

  1. Serological diagnosis of avian influenza in poultry

    DEFF Research Database (Denmark)

    Comin, Arianna; Toft, Nils; Stegeman, Arjan;

    2013-01-01

    Background The serological diagnosis of avian influenza (AI) can be performed using different methods, yet the haemagglutination inhibition (HI) test is considered the gold standard' for AI antibody subtyping. Although alternative diagnostic assays have been developed, in most cases, their accuracy...

  2. Measuring steroid hormones in avian eggs

    NARCIS (Netherlands)

    Von Engelhardt, N; Groothuis, TGG; Bauchinger, U; Goymann, W; JenniEiermann, S

    2005-01-01

    Avian eggs contain substantial levels of various hormones of maternal origin and have recently received a lot of interest, mainly from behavioral ecologists. These studies strongly depend on the measurement of egg hormone levels, but the method of measuring these levels has received little attention

  3. Avian Disease & Oncology Lab (ADOL) Research Update

    Science.gov (United States)

    Employing Genomics, Epigenetics, and Immunogenetics to Control Diseases Induced by Avian Tumor Viruses - Gene expression is a major factor accounting for phenotypic variation. Taking advantage of allele-specific expression (ASE) screens, we found the use of genetic markers was superior to traditiona...

  4. Fossil avian eggshell preserves ancient DNA

    DEFF Research Database (Denmark)

    Oskam, Charlotte L; Haile, James Seymour; McLay, Emma

    2010-01-01

    Owing to exceptional biomolecule preservation, fossil avian eggshell has been used extensively in geochronology and palaeodietary studies. Here, we show, to our knowledge, for the first time that fossil eggshell is a previously unrecognized source of ancient DNA (aDNA). We describe the successful...

  5. Measuring Steroid Hormones in Avian Eggs

    NARCIS (Netherlands)

    Engelhardt, Nikolaus von; Groothuis, Ton G.G.

    2005-01-01

    Avian eggs contain substantial levels of various hormones of maternal origin and have recently received a lot of interest, mainly from behavioral ecologists. These studies strongly depend on the measurement of egg hormone levels, but the method of measuring these levels has received little attention

  6. Website for avian flu information and bioinformatics

    Institute of Scientific and Technical Information of China (English)

    GAO; George; Fu

    2009-01-01

    Highly pathogenic influenza A virus H5N1 has spread out worldwide and raised the public concerns. This increased the output of influenza virus sequence data as well as the research publication and other reports. In order to fight against H5N1 avian flu in a comprehensive way, we designed and started to set up the Website for Avian Flu Information (http://www.avian-flu.info) from 2004. Other than the influenza virus database available, the website is aiming to integrate diversified information for both researchers and the public. From 2004 to 2009, we collected information from all aspects, i.e. reports of outbreaks, scientific publications and editorials, policies for prevention, medicines and vaccines, clinic and diagnosis. Except for publications, all information is in Chinese. Till April 15, 2009, the cumulative news entries had been over 2000 and research papers were approaching 5000. By using the curated data from Influenza Virus Resource, we have set up an influenza virus sequence database and a bioinformatic platform, providing the basic functions for the sequence analysis of influenza virus. We will focus on the collection of experimental data and results as well as the integration of the data from the geological information system and avian influenza epidemiology.

  7. [Avian influenza and oseltamivir; a retrospective view

    NARCIS (Netherlands)

    Galama, J.M.D.

    2003-01-01

    The outbreak of avian influenza A due to an H7N7 virus in Dutch poultry farms turned out to have public-health effects for those who were involved in the management of the epidemic and who were thus extensively exposed to contaminated excreta and dust. An outbreak-management team (OMT) of experts in

  8. Website for avian flu information and bioinformatics

    Institute of Scientific and Technical Information of China (English)

    LIU Di; LIU Quan-He; WU Lin-Huan; LIU Bin; WU Jun; LAO Yi-Mei; LI Xiao-Jing; GAO George Fu; MA Jun-Cai

    2009-01-01

    Highly pathogenic influenza A virus H5N1 has spread out worldwide and raised the public concerns. This increased the output of influenza virus sequence data as well as the research publication and other reports. In order to fight against H5N1 avian flu in a comprehensive way, we designed and started to set up the Website for Avian Flu Information (http://www.avian-flu.info) from 2004. Other than the influenza virus database available, the website is aiming to integrate diversified information for both researchers and the public. From 2004 to 2009, we collected information from all aspects, i.e. reports of outbreaks, scientific publications and editorials, policies for prevention, medicines and vaccines, clinic and diagnosis. Except for publications, all information is in Chinese. Till April 15, 2009, the cumulative news entries had been over 2000 and research papers were approaching 5000. By using the curated data from Influenza Virus Resource, we have set up an influenza virus sequence database and a bioin-formatic platform, providing the basic functions for the sequence analysis of influenza virus. We will focus on the collection of experimental data and results as well as the integration of the data from the geological information system and avian influenza epidemiology.

  9. Activation of AMPKα2 is not crucial for mitochondrial uncoupling-induced metabolic effects but required to maintain skeletal muscle integrity.

    Directory of Open Access Journals (Sweden)

    Mario Ost

    Full Text Available Transgenic (UCP1-TG mice with ectopic expression of UCP1 in skeletal muscle (SM show a phenotype of increased energy expenditure, improved glucose tolerance and increase substrate metabolism in SM. To investigate the potential role of skeletal muscle AMPKα2 activation in the metabolic phenotype of UCP1-TG mice we generated double transgenic (DTG mice, by crossing of UCP1-TG mice with DN-AMPKα2 mice overexpressing a dominant negative α2 subunit of AMPK in SM which resulted in an impaired AMPKα2 activity by 90±9% in SM of DTG mice. Biometric analysis of young male mice showed decreased body weight, lean and fat mass for both UCP1-TG and DTG compared to WT and DN-AMPKα2 mice. Energy intake and weight-specific total energy expenditure were increased, both in UCP1-TG and DTG mice. Moreover, glucose tolerance, insulin sensitivity and fatty acid oxidation were not altered in DTG compared to UCP1-TG. Also uncoupling induced induction and secretion of fibroblast growth factor 21 (FGF21 from SM was preserved in DTG mice. However, voluntary physical cage activity as well as ad libitum running wheel access during night uncovered a severe activity intolerance of DTG mice. Histological analysis showed a progressive degenerative morphology in SM of DTG mice which was not observed in SM of UCP1-TG mice. Moreover, ATP-depletion related cellular stress response via heat shock protein 70 was highly induced, whereas capillarization regulator VEGF was suppressed in DTG muscle. In addition, AMPKα2-mediated induction of mitophagy regulator ULK1 was suppressed in DTG mice, as well as mitochondrial respiratory capacity and content. In conclusion, we demonstrate that AMPKα2 is dispensable for SM mitochondrial uncoupling induced metabolic effects on whole body energy balance, glucose homeostasis and insulin sensitivity. But strikingly, activation of AMPKα2 seems crucial for maintaining SM function, integrity and the ability to compensate chronic metabolic stress

  10. Dual function of the hemagglutinin H5 fused to chicken CD154 in a potential strategy of DIVA against avian influenza disease: preliminary study

    Science.gov (United States)

    Pose, A.G.; Rodríguez, E.S.; Méndez, A.C.; Gómez, J.N.; Redondo, A.V.; Rodríguez, E.R.; Ramos, E.M.G.; Gutiérrez, A.Á.; Moltó, M.P.R.; Roche, D.G.; Ugalde, Y.S.; López, A.M.

    2015-01-01

    In this study we demonstrated that the vaccine candidate against avian influenza virus H5N1 based on the hemagglutinin H5 (HA) fused to the chicken CD154 (HACD) can also be used for differentiating infected from vaccinated animals (DIVA). As the strategy of DIVA requires at least two proteins, we obtained a variant of the nucleoprotein (NP49-375) in E. coli. After its purification by IMAC, the competence of the proteins NP49-375 and HACD as coating antigens in indirect ELISA assays were tested by using the sera of chickens immunized with the proteins HA and HACD and the reference sera from several avian influenza subtypes. Together with these sera, the sera from different species of birds and the sera of chickens infected with other avian viral diseases were analyzed by competition ELISA assays coated with the proteins NP49-375 and HACD. The results showed that the segment CD154 in the chimeric protein HACD did not interfere with the recognition of the molecule HA by its specific antibodies. Also, we observed variable detection levels when the reference sera were analyzed in the ELISA plates coated with the protein NP49-375. Moreover, only the antibodies of the reference serum subtype H5 were detected in the ELISA plates coated with the protein HACD. The competition ELISA assays showed percentages of inhibition of 88-91% for the positives sera and less than 20% for the negative sera. We fixed the cut-off value of these assays at 25%. No antibody detection was observed in the sera from different species of birds or the sera of chickens infected with other avian viral diseases. This study supported the fact that the ELISA assays using the proteins NP49-375 and HACD could be valuable tools for avian influenza surveillance and as a strategy of DIVA for counteracting the highly pathogenic avian influenza virus H5N1 outbreaks. PMID:26623380

  11. Expression analysis of TALE family transcription factors during avian development.

    Science.gov (United States)

    Coy, Sarah E; Borycki, Anne-Gaëlle

    2010-04-01

    The TALE family of homeodomain containing transcription factors consists of the Meis, Prep and Tgif, and the Pbx subfamily of proteins. Several TALE orthologues have been identified in amniotes, but no comprehensive analysis of their expression pattern during embryogenesis has been performed. Here, we report on TALE gene expression in the avian embryo. During embryonic development, Pbx genes are predominantly expressed in the neural ectoderm and paraxial mesoderm, although Pbx3 is restricted to the intermediate and lateral mesoderm, and anterior central nervous system. Members of the Meis, Prep, and Tgif subfamilies are expressed at high levels in the paraxial mesoderm, and display differential expression along the anterior-posterior and dorsoventral axes of the developing neural tube. Overall the expression patterns reported in this study are consistent with the known function of the TALE gene family in controlling early patterning of limb, neural tube and paraxial mesoderm tissues during embryogenesis.

  12. Molecular detection of avian pox virus from nodular skin and mucosal fibrinonecrotic lesions of Iranian backyard poultry.

    Science.gov (United States)

    Gholami-Ahangaran, Majid; Zia-Jahromi, Noosha; Namjoo, Abdolrasul

    2014-02-01

    In recent years, some outbreaks of skin lesions suspected to be avian pox were observed in the backyard poultry in different parts of western areas in Iran. Consequently, 328 backyard poultries with suspected signs of avian pox virus infection were sampled. All birds showed nodular lesions on unfeathered head skin and/or fibronecrotic lesions on mucus membrane of the oral cavity and upper respiratory tract. For histopathological analysis, the sections of tissue samples from cutaneous lesions of examined birds were stained with H&E method. For PCR, after DNA extraction a 578-bp fragment of avian pox virus from 4b core protein gene was amplified. Results showed 217 and 265 out of 328 (66.1 and 80.7%, respectively) samples were positive for avian pox virus on histopathological and PCR examination, respectively. In this study, the samples that had intracytoplasmic inclusion bodies on pathologic examination were PCR positive. This study revealed that PCR is a valuable tool for identification of an avian pox virus and that the frequency of pox infection in backyard poultry in western areas of Iran is high.

  13. gga-miR-375 plays a key role in tumorigenesis post subgroup J avian leukosis virus infection.

    Science.gov (United States)

    Li, Hongxin; Shang, Huiqing; Shu, Dingming; Zhang, Huanmin; Ji, Jun; Sun, Baoli; Li, Hongmei; Xie, Qingmei

    2014-01-01

    Avian leukosis is a neoplastic disease caused in part by subgroup J avian leukosis virus J (ALV-J). Micro ribonucleic acids (miRNAs) play pivotal oncogenic and tumour-suppressor roles in tumour development and progression. However, little is known about the potential role of miRNAs in avian leukosis tumours. We have found a novel tumour-suppressor miRNA, gga-miR-375, associated with avian leukosis tumorigenesis by miRNA microarray in a previous report. We have also previously studied the biological function of gga-miR-375; Overexpression of gga-miR-375 significantly inhibited DF-1 cell proliferation, and significantly reduced the expression of yes-associated protein 1 (YAP1) by repressing the activity of a luciferase reporter carrying the 3'-untranslated region of YAP1. This indicates that gga-miR-375 is frequently downregulated in avian leukosis by inhibiting cell proliferation through YAP1 oncogene targeting. Overexpression of gga-miR-375 markedly promoted serum starvation induced apoptosis, and there may be the reason why the tumour cycle is so long in the infected chickens. In vivo assays, gga-miR-375 was significantly downregulated in chicken livers 20 days after infection with ALV-J, and YAP1 was significantly upregulated 20 days after ALV-J infection (Pleukosis tumorigenesis.

  14. Novel Polymerase Gene Mutations for Human Adaptation in Clinical Isolates of Avian H5N1 Influenza Viruses.

    Science.gov (United States)

    Arai, Yasuha; Kawashita, Norihito; Daidoji, Tomo; Ibrahim, Madiha S; El-Gendy, Emad M; Takagi, Tatsuya; Takahashi, Kazuo; Suzuki, Yasuo; Ikuta, Kazuyoshi; Nakaya, Takaaki; Shioda, Tatsuo; Watanabe, Yohei

    2016-04-01

    A major determinant in the change of the avian influenza virus host range to humans is the E627K substitution in the PB2 polymerase protein. However, the polymerase activity of avian influenza viruses with a single PB2-E627K mutation is still lower than that of seasonal human influenza viruses, implying that avian viruses require polymerase mutations in addition to PB2-627K for human adaptation. Here, we used a database search of H5N1 clade 2.2.1 virus sequences with the PB2-627K mutation to identify other polymerase adaptation mutations that have been selected in infected patients. Several of the mutations identified acted cooperatively with PB2-627K to increase viral growth in human airway epithelial cells and mouse lungs. These mutations were in multiple domains of the polymerase complex other than the PB2-627 domain, highlighting a complicated avian-to-human adaptation pathway of avian influenza viruses. Thus, H5N1 viruses could rapidly acquire multiple polymerase mutations that function cooperatively with PB2-627K in infected patients for optimal human adaptation.

  15. Explicit thin-lens solution for an arbitrary four by four uncoupled beam transfer matrix

    Energy Technology Data Exchange (ETDEWEB)

    Balandin, V. [Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Orlov, S. [Moscow State Univ. (Russian Federation). Faculty of Computational Mathematics and Cybernetics

    2011-10-15

    In the design of beam transport lines one often meets the problem of constructing a quadrupole lens system that will produce desired transfer matrices in both the horizontal and vertical planes. Nowadays this problem is typically approached with the help of computer routines, but searching for the numerical solution one has to remember that it is not proven yet that an arbitrary four by four uncoupled beam transfer matrix can be represented by using a finite number of drifts and quadrupoles (representation problem) and the answer to this questions is not known not only for more or less realistic quadrupole field models but also for the both most commonly used approximations of quadrupole focusing, namely thick and thin quadrupole lenses. In this paper we make a step forward in resolving the representation problem and, by giving an explicit solution, we prove that an arbitrary four by four uncoupled beam transfer matrix actually can be obtained as a product of a finite number of thin-lenses and drifts. (orig.)

  16. Divergent roles for Eph and Ephrin in Avian Cranial Neural Crest

    Directory of Open Access Journals (Sweden)

    Burke Robert D

    2008-05-01

    Full Text Available Abstract Background As in other vertebrates, avian hindbrain neural crest migrates in streams to specific branchial arches. Signalling from Eph receptors and ephrins has been proposed to provide a molecular mechanism that guides the cells restricting them to streams. In mice and frogs, cranial neural crest express a combination of Eph receptors and ephrins that appear to exclude cells from adjacent tissues by forward and reverse signalling. The objective of this study was to provide comparative data on the distribution and function of Eph receptors and ephrins in avian embryos. Results To distinguish neural crest from bordering ectoderm and head mesenchyme, we have co-labelled embryos for Eph or ephrin RNA and a neural crest marker protein. Throughout their migration avian cranial neural crest cells express EphA3, EphA4, EphA7, EphB1, and EphB3 and move along pathways bordered by non-neural crest cells expressing ephrin-B1. In addition, avian cranial neural crest cells express ephrin-B2 and migrate along pathways bordered by non-neural crest cells expressing EphB2. Thus, the distribution of avian Eph receptors and ephrins differs from those reported in other vertebrates. In stripe assays when explanted cranial neural crest were given the choice between FN or FN plus clustered ephrin-B1 or EphB2 fusion protein, the cells strongly localize to lanes containing only FN. This preference is mitigated in the presence of soluble ephrin-B1 or EphB2 fusion protein. Conclusion These findings show that avian cranial neural crest use Eph and ephrin receptors as other vertebrates in guiding migration. However, the Eph receptors are expressed in different combinations by neural crest destined for each branchial arch and ephrin-B1 and ephrin-B2 appear to have opposite roles to those reported to guide cranial neural crest migration in mice. Unlike many of the signalling, specification, and effector pathways of neural crest, the roles of Eph receptors and ephrins

  17. Activation of mitochondrial ATP-sensitive potassium channels delays ischemia-induced cellular uncoupling in rat heart

    Institute of Scientific and Technical Information of China (English)

    Yue-liangSHEN; Ying-yingCHEN; Xun-dongWU; IainCBRUCE; QiangXIA

    2004-01-01

    AIM: To test the hypothesis that cellular uncoupling induced by myocardial ischemia is mediated by activation of mitochondrial ATP-sensitive potassium channels (mitoKATP). METHODS: Rat hearts were perfused on a Langendorff apparatus and subjected to 40-min ischemia followed by 30-min reperfusion (I/R). Changes in cellular coupling were monitored by measuring whole-tissue resistance. RESULTS: (1) In hearts subjected to I/R, the onset of uncoupling started at (13.3± 1.0) min of ischemia; (2) Ischemic preconditioning (IPC) delayed the onset of uncoupling until (22.7±1.3) min. Blocking mitoKATP channels with 5-hydroxydecanoate (5-HD) before the IPC abolishedthe uncoupling delay [(12.6±1.6)min]; (3) Calcium preconditioning (CPC) had the same effect as IPC. And this effect was reversed by blocking the mitoKATP channel again. In the CPC group the onset of uncoupling occurred after (20.6±1.3) min, and this was canceled by 5-HD [(13.6±0.8) min]; (4) In hearts pretreated with the specific mitoKATP channel opener diazoxide before sustained ischemia, the onset was delayed to (18.4±1.4) min; (5) 5-HD canceled the protective effects of diazoxide (12.6±1.0) min; and both the L-type Ca2+ channel inhibitor verapamiland the free radical scavenger N-(2-mercaptopropionyl)glycine, reduced the extended onset time induced by diazoxide[to (13.3±1.8) min and (13.4±2.1) min, respectively]. CONCLUSION: IPC and CPC delay the onset of cellular uncoupling induced by acute ischemia in rat heart, and the underlying mechanism involves activation of the mitoKATP channels.

  18. gga-miR-375 plays a key role in tumorigenesis post subgroup J avian leukosis virus infection.

    Directory of Open Access Journals (Sweden)

    Hongxin Li

    Full Text Available Avian leukosis is a neoplastic disease caused in part by subgroup J avian leukosis virus J (ALV-J. Micro ribonucleic acids (miRNAs play pivotal oncogenic and tumour-suppressor roles in tumour development and progression. However, little is known about the potential role of miRNAs in avian leukosis tumours. We have found a novel tumour-suppressor miRNA, gga-miR-375, associated with avian leukosis tumorigenesis by miRNA microarray in a previous report. We have also previously studied the biological function of gga-miR-375; Overexpression of gga-miR-375 significantly inhibited DF-1 cell proliferation, and significantly reduced the expression of yes-associated protein 1 (YAP1 by repressing the activity of a luciferase reporter carrying the 3'-untranslated region of YAP1. This indicates that gga-miR-375 is frequently downregulated in avian leukosis by inhibiting cell proliferation through YAP1 oncogene targeting. Overexpression of gga-miR-375 markedly promoted serum starvation induced apoptosis, and there may be the reason why the tumour cycle is so long in the infected chickens. In vivo assays, gga-miR-375 was significantly downregulated in chicken livers 20 days after infection with ALV-J, and YAP1 was significantly upregulated 20 days after ALV-J infection (P<0.05. We also found that expression of cyclin E, an important regulator of cell cycle progression, was significantly upregulated (P<0.05. Drosophila inhibitor of apoptosis protein 1 (DIAP1, which is related to caspase-dependent apoptosis, was also significantly upregulated after infection. Our data suggests that gga-miR-375 may function as a tumour suppressor thereby regulating cancer cell proliferation and it plays a key role in avian leukosis tumorigenesis.

  19. Research update: Avian Disease and Oncology Laboratory avian tumor viruses

    Science.gov (United States)

    Genomics and Immunogenetics Use of genomics to identify QTL, genes, and proteins associated with resistance to Marek’s disease. Marek’s disease (MD), a lymphoproliferative disease caused by the highly oncogenic herpesvirus Marek's disease virus (MDV), continues to be a major disease concern to the p...

  20. The N Terminus of Sarcolipin Plays an Important Role in Uncoupling Sarco-endoplasmic Reticulum Ca2+-ATPase (SERCA) ATP Hydrolysis from Ca2+ Transport

    DEFF Research Database (Denmark)

    Sahoo, Sanjaya K; Shaikh, Sana A; Sopariwala, Danesh H;

    2015-01-01

    to bind SERCA throughout its kinetic cycle and promotes uncoupling of Ca(2+) transport from ATP hydrolysis. To determine the structural regions of SLN that mediate uncoupling of SERCA, we employed mutagenesis and generated chimeras of PLB and SLN. In this study we demonstrate that deletion of SLN N...

  1. Deteksi Antibodi Serum Terhadap Virus Avian influenza pada Ayam Buras

    Directory of Open Access Journals (Sweden)

    Darmawi Darmawi

    2012-04-01

    Full Text Available Detection on Serum Antibodies of Native Chickens to Avian influenza Virus ABSTRACT.  An important approach of controlling against Avian Influenza should be determined to detect the antibody titres of bird flu caused by Influenza virus H5N1 in Indonesia. The aim of the present study was to detect the antibodies to Avian Influenza in serum of native chickens. This study utilized 123 serum samples collected from the axilaris vein (left or right of native chickens. Antibody titres were examined using Hemaglutination Inhibition (HI. The result showed that indication of natural infection by Avian Influenza (H5N1 in native chickens, as shown that out of 123 serum samples, 16 (13,01% were tested positive by HI, while only 10 (8,13% were tested protective to Avian influenza infection. Based on the results we obtained, a conclusion that natural infection by Avian influenza virus stimulated variety level of formation antibody titres in native chickens.

  2. H5N1禽流感病毒NS1蛋白与干扰素诱导蛋白10表达的相关性研究%Influence of avian influenza virus NS1 protein on the expression of IP-10 in BEAS-2B cells

    Institute of Scientific and Technical Information of China (English)

    贾晓俊; 周剑芳; 王晶钰; 董婕; 薄洪; 李梓; 李魁彪; 蓝雨; 舒跃龙

    2008-01-01

    目的 研究高致病性禽流感(HPAI)H5N1病毒NS1蛋白对干扰素诱导蛋白10(IP-10)的影响.方法 分别将禽流感病毒A/Anhui/1/2005(H5N1)的NS1基因、插入80-84位缺失氨基酸的NS1突变基因及流感病毒A/Puerto Rico/8/1934(H1N1)的NS1基因克隆至真核表达载体pEGFP-N1,转染人支气管上皮细胞BEAS-2B,流式细胞仪检测转染细胞内IP-10的表达情况.结果 与pEGFP-N1对照组相比,三种NS1蛋白均能下调BEAS-2B细胞IP-10的表达(P0.01).结论 A/Anhui/1/2005(H5N1)禽流感病毒单一NS1蛋白能够抑制BEAS-2B细胞IP-10表达,但这并不能完全阐明其与病毒致病性之间的关系.%Objective To investigate the influence of avian influenza virus (AIV) NS1 protein on the expression of interferon-inducible protein 10 (IP-10). Methods NS1 gene from virus A/Anhui/1/2005 (H5N1),NS1 gene inserted with 80-84 amino acids from virus A/Anhui/1/2005(H5N1)and NS1 gene from virus A/Puerto Rico/8/1934 (H1N1) were cloned into the eukaryotic expression vector pEGFP-N1, and transected into BEAS-2Bcells, IP-10 expression level in transected cells was detected by flow cytometry. Results Compared with the control group pEGFP-N1, Expression of these three different NS1 genes can down-regulate the expression of IP-10in BEAS-2B cells, but there is no significant difference as to the lower level among them. Conclusion NS1protein of A/Anhui/1/2005(H5N1) can down-regulate the expression level of IP-10, but this may not clarify its relationship with the virulence of AIV.

  3. Avian use of Norris Hill Wind Resource Area, Montana

    Energy Technology Data Exchange (ETDEWEB)

    Harmata, A.; Podruzny, K.; Zelenak, J. [Montana State Univ., Bozeman, MT (United States). Biology Dept.

    1998-07-01

    This document presents results of a study of avian use and mortality in and near a proposed wind resource area in southwestern Montana. Data collected in autumn 1995 through summer 1996 represented preconstruction condition; it was compiled, analyzed, and presented in a format such that comparison with post-construction data would be possible. The primary emphasis of the study was recording avian migration in and near the wind resource area using state-of-the-art marine surveillance radar. Avian use and mortality were investigated during the breeding season by employing traditional avian sampling methods, radiotelemetry, radar, and direct visual observation. 61 figs., 34 tabs.

  4. Avian influenza virus and free-ranging wild birds

    Science.gov (United States)

    Dierauf, Leslie A.; Karesh, W.B.; Ip, Hon S.; Gilardi, K.V.; Fischer, John R.

    2006-01-01

    Recent media and news reports and other information implicate wild birds in the spread of highly pathogenic avian influenza in Asia and Eastern Europe. Although there is little information concerning highly pathogenic avian influenza viruses in wild birds, scientists have amassed a large amount of data on low-pathogenicity avian influenza viruses during decades of research with wild birds. This knowledge can provide sound guidance to veterinarians, public health professionals, the general public, government agencies, and other entities with concerns about avian influenza.

  5. Development and characterization of monoclonal antibodies to subgroup A avian leukosis virus.

    Science.gov (United States)

    Qiu, Y; Li, X; Fu, L; Cui, Z; Li, W; Wu, Z; Sun, S

    2014-03-01

    Avian leukosis virus subgroup A (ALV-A) is a retrovirus which infects egg-type chickens and is the main pathogen of lymphoid leukosis (LL) and myeloid leukosis (ML). In order to greatly enhance the diagnosis and treatment of clinical avian leukemia, two monoclonal antibodies (MAbs) to ALV-A were developed by fusion between SP2/0 and spleen cells from mice immunized with expressed ALV-A env-gp85 protein. Using immunofluorescence assay (IFA), two MAbs reacted with ALV-A, but not with subgroups B and J of ALV. Western blot tests showed that molecular weight of ALV-A envelope glycoprotein recognized by MAbs was about 53 kD. Isotyping test revealed that two MAbs (A5C1 and A4C8) were IgG1 isotypes. These MAbs can be used for diagnosis and epidemiology of ALV-A.

  6. Interspecies transmission and host restriction of avian H5N1 influenza virus

    Institute of Scientific and Technical Information of China (English)

    LIU Di; LIU XiaoLing; YAN JingHua; LIU Wen-Jun; GAO George Fu

    2009-01-01

    Long-term endemicity of avian H5N1 influenza virus in poultry and continuous sporadic human infec-tions in several countries has raised the concern of another potential pandemic influenza. Suspicion of the avian origin of the previous pandemics results in the close investigation of the mechanism of in-terspecies transmission. Entry and fusion is the first step for the H5N1 influenza virus to get into the host cells affecting the host ranges. Therefore receptor usage study has been a major focus for the last few years. We now know the difference of the sialic acid structures and distributions in different spe-cies, even in the different parts of the same host. Many host factors interacting with the influenza virus component proteins have been identified and their role in the host range expansion and interspecies transmission is under detailed scrutiny. Here we review current progress in the receptor usage and host factors.

  7. Interspecies transmission and host restriction of avian H5N1 influenza virus

    Institute of Scientific and Technical Information of China (English)

    GAO; George; Fu

    2009-01-01

    Long-term endemicity of avian H5N1 influenza virus in poultry and continuous sporadic human infections in several countries has raised the concern of another potential pandemic influenza. Suspicion of the avian origin of the previous pandemics results in the close investigation of the mechanism of interspecies transmission. Entry and fusion is the first step for the H5N1 influenza virus to get into the host cells affecting the host ranges. Therefore receptor usage study has been a major focus for the last few years. We now know the difference of the sialic acid structures and distributions in different species, even in the different parts of the same host. Many host factors interacting with the influenza virus component proteins have been identified and their role in the host range expansion and interspecies transmission is under detailed scrutiny. Here we review current progress in the receptor usage and host factors.

  8. Seasonal change in the avian hippocampus.

    Science.gov (United States)

    Sherry, David F; MacDougall-Shackleton, Scott A

    2015-04-01

    The hippocampus plays an important role in cognitive processes, including memory and spatial orientation, in birds. The hippocampus undergoes seasonal change in food-storing birds and brood parasites, there are changes in the hippocampus during breeding, and further changes occur in some species in association with migration. In food-storing birds, seasonal change in the hippocampus occurs in fall and winter when the cognitively demanding behaviour of caching and retrieving food occurs. The timing of annual change in the hippocampus of food-storing birds is quite variable, however, and appears not to be under photoperiod control. A variety of factors, including cognitive performance, exercise, and stress may all influence seasonal change in the avian hippocampus. The causal processes underlying seasonal change in the avian hippocampus have not been extensively examined and the more fully described hormonal influences on the mammalian hippocampus may provide hypotheses for investigating the control of hippocampal seasonality in birds.

  9. Two novel neutralizing antigenic epitopes of the s1 subunit protein of a QX-like avian infectious bronchitis virus strain Sczy3 as revealed using a phage display peptide library.

    Science.gov (United States)

    Zou, Nianli; Xia, Jing; Wang, Fuyan; Duan, Zhenzhen; Miao, Dan; Yan, Qigui; Cao, Sanjie; Wen, Xintian; Liu, Ping; Huang, Yong

    2015-11-15

    The spike (S) protein of the infectious bronchitis virus (IBV) plays a central role in the pathogenicity, the immune antibody production, serotype and the tissue tropism. In this study, we generate 11 monoclonal antibodies (mAbs) against S1 subunit of IBV Sczy3 strain, and two mAbs 1D5 and 6A12 were positive in indirect ELISA against both His-S1 protein and the purified whole viral antigen. MAb 6A12 and 1D5 could recognized by other 10 IBV strains (IBVs) from five different genotypes, except that 1D5 had a relatively low reaction with two of the 10 tested IBVs. End-point neutralizing assay performed in chicken embro kidney (CEK) cells revealed that the neutralization titer of 6A12 and 1D5 against Sczy3 reached 1:44.7 and 1:40.6, respectively. After screening a phage display peptide library and peptide scanning, we identified two linear B-cell epitopes that were recognized by the mAbs 1D5 and 6A12, which corresponded to the amino acid sequences (87)PPQGMAW(93) and (412)IQTRTEP(418), respectively, in the IBV S1 subunit. Sequences comparison revealed that epitope (412)IQTRTEP(418) was conserved among IBVs, while the epitope (87)PPQGMAW(93) was relatively variable among IBVs. The novel mAbs and the epitopes identified will be useful for developing diagnostic assays for IBV infections.

  10. Avian Interferons and Their Antiviral Effectors

    OpenAIRE

    Santhakumar, Diwakar; Rubbenstroth, Dennis; Martinez-Sobrido, Luis; Munir, Muhammad

    2017-01-01

    Interferon (IFN) responses, mediated by a myriad of IFN-stimulated genes (ISGs), are the most profound innate immune responses against viruses. Cumulatively, these IFN effectors establish a multilayered antiviral state to safeguard the host against invading viral pathogens. Considerable genetic and functional characterizations of mammalian IFNs and their effectors have been made, and our understanding on the avian IFNs has started to expand. Similar to mammalian counterparts, three types of I...

  11. Mitochondrial uncouplers act synergistically with the fumigant phosphine to disrupt mitochondrial membrane potential and cause cell death.

    Science.gov (United States)

    Valmas, Nicholas; Zuryn, Steven; Ebert, Paul R

    2008-10-30

    Phosphine is the most widely used fumigant for the protection of stored commodities against insect pests, especially food products such as grain. However, pest insects are developing resistance to phosphine and thereby threatening its future use. As phosphine inhibits cytochrome c oxidase (complex IV) of the mitochondrial respiratory chain and reduces the strength of the mitochondrial membrane potential (DeltaPsi(m)), we reasoned that mitochondrial uncouplers should act synergistically with phosphine. The mitochondrial uncouplers FCCP and PCP caused complete mortality in populations of both wild-type and phosphine-resistant lines of Caenorhabditis elegans simultaneously exposed to uncoupler and phosphine at concentrations that were individually nonlethal. Strong synergism was also observed with a third uncoupler DNP. We have also tested an alternative complex IV inhibitor, azide, with FCCP and found that this also caused a synergistic enhancement of toxicity in C. elegans. To investigate potential causes of the synergism, we measured DeltaPsi(m), ATP content, and oxidative damage (lipid hydroperoxides) in nematodes subjected to phosphine-FCCP treatment and found that neither an observed 50% depletion in ATP nor oxidative stress accounted for the synergistic effect. Instead, a synergistic reduction in DeltaPsi(m) was observed upon phosphine-FCCP co-treatment suggesting that this is directly responsible for the subsequent mortality. These results support the hypothesis that phosphine-induced mortality results from the in vivo disruption of normal mitochondrial activity. Furthermore, we have identified a novel pathway that can be targeted to overcome genetic resistance to phosphine.

  12. The avian fossil record in Insular Southeast Asia and its implications for avian biogeography and palaeoecology.

    Science.gov (United States)

    Meijer, Hanneke J M

    2014-01-01

    Excavations and studies of existing collections during the last decades have significantly increased the abundance as well as the diversity of the avian fossil record for Insular Southeast Asia. The avian fossil record covers the Eocene through the Holocene, with the majority of bird fossils Pleistocene in age. Fossil bird skeletal remains represent at least 63 species in 54 genera and 27 families, and two ichnospecies are represented by fossil footprints. Birds of prey, owls and swiftlets are common elements. Extinctions seem to have been few, suggesting continuity of avian lineages since at least the Late Pleistocene, although some shifts in species ranges have occurred in response to climatic change. Similarities between the Late Pleistocene avifaunas of Flores and Java suggest a dispersal route across southern Sundaland. Late Pleistocene assemblages of Niah Cave (Borneo) and Liang Bua (Flores) support the rainforest refugium hypothesis in Southeast Asia as they indicate the persistence of forest cover, at least locally, throughout the Late Pleistocene and Holocene.

  13. Seroprevalence of avian hepatitis E virus and avian leucosis virus subgroup J in chicken flocks with hepatitis syndrome, China

    OpenAIRE

    Sun, Yani; Du, Taofeng; Liu, Baoyuan; Syed, Shahid Faraz; Chen, Yiyang; Li, Huixia; Wang, Xinjie; Zhang, Gaiping; Zhou, En-Min; Zhao, Qin

    2016-01-01

    Background From 2014 to 2015 in China, many broiler breeder and layer hen flocks exhibited a decrease in egg production and some chickens developed hepatitis syndrome including hepatomegaly, hepatic necrosis and hemorrhage. Avian hepatitis E virus (HEV) and avian leucosis virus subgroup J (ALV-J) both cause decreasing in egg production, hepatomegaly and hepatic hemorrhage in broiler breeder and layer hens. In the study, the seroprevalence of avian HEV and ALV-J in these flocks emerging the di...

  14. Spatial uncoupling of biodegradation, soil respiration, and PAH concentration in a creosote contaminated soil.

    Science.gov (United States)

    Bengtsson, Göran; Törneman, Niklas; Yang, Xiuhong

    2010-09-01

    Hotspots and coldspots of concentration and biodegradation of polycyclic aromatic hydrocarbons (PAHs) marginally overlapped at the 0.5-100 m scale in a creosote contaminated soil in southern Sweden, suggesting that concentration and biodegradation had little spatial co-variation. Biodegradation was substantial and its spatial variability considerable and highly irregular, but it had no spatial autocorrelation. The soil concentration of PAHs explained only 20-30% of the variance of their biodegradation. Soil respiration was spatially autocorrelated. The spatial uncoupling between biodegradation and soil respiration seemed to be governed by the aging of PAHs in the soil, since biodegradation of added 13C phenanthrene covaried with both soil respiration and microbial biomass. The latter two were also correlated with high concentrations of phospholipid fatty acids (PLFAs) that are common in gram-negative bacteria. However, several of the hotspots of biodegradation coincided with hotspots for the distribution of a PLFA indicative of fungal biomass.

  15. Detection of Mechanism of Noise-Induced Synchronization between Two Identical Uncoupled Neurons

    Institute of Scientific and Technical Information of China (English)

    WU Ying; XU Jian-Xue; JIN Wu-Yin; HONG Ling

    2007-01-01

    We investigate the noise-induced synchronization between two identical uncoupled Hodgkin-Huxley neurons with sinusoidal stimulations. The numerical results confirm that the value of critical noise intensity for synchronizing two systems is much less than the magnitude of mean size of the attractor in the original system, and the deterministic feature of the attractor in the original system remains unchanged. This finding is significantly different from the previous work [Phys. Rev. E 67 (2003) 027201] in which the value of the critical noise intensity for synchronizing two systems was found to be roughly equal to the magnitude of mean size of the attractor in the original system, and at this intensity, the noise swamps the qualitative structure of the attractor in the original deterministic systems to synchronize to their stochastic dynamics. Further investigation shows that the critical noise intensity for synchronizing two neurons induced by noise may be related to the structure of interspike intervals of the original systems.

  16. Mild mitochondrial uncoupling and calorie restriction increase fasting eNOS, akt and mitochondrial biogenesis.

    Directory of Open Access Journals (Sweden)

    Fernanda M Cerqueira

    Full Text Available Enhanced mitochondrial biogenesis promoted by eNOS activation is believed to play a central role in the beneficial effects of calorie restriction (CR. Since treatment of mice with dinitrophenol (DNP promotes health and lifespan benefits similar to those observed in CR, we hypothesized that it could also impact biogenesis. We found that DNP and CR increase citrate synthase activity, PGC-1α, cytochrome c oxidase and mitofusin-2 expression, as well as fasting plasma levels of NO• products. In addition, eNOS and Akt phosphorylation in skeletal muscle and visceral adipose tissue was activated in fasting CR and DNP animals. Overall, our results indicate that systemic mild uncoupling activates eNOS and Akt-dependent pathways leading to mitochondrial biogenesis.

  17. Variable ATP yields and uncoupling of oxygen consumption in human brain

    DEFF Research Database (Denmark)

    Gjedde, Albert; Aanerud, Joel; Peterson, Ericka;

    2011-01-01

    The distribution of brain oxidative metabolism values among healthy humans is astoundingly wide for a measure that reflects normal brain function and is known to change very little with most changes of brain function. It is possible that the part of the oxygen consumption rate that is coupled...... to ATP turnover is the same in all healthy human brains, with different degrees of uncoupling explaining the variability of total oxygen consumption among people. To test the hypothesis that about 75% of the average total oxygen consumption of human brains is common to all individuals, we determined...... the variability in a large group of normal healthy adults. To establish the degree of variability in different regions of the brain, we measured the regional cerebral metabolic rate for oxygen in 50 healthy volunteers aged 21-66 and projected the values to a common age of 25.Within each subject and region, we...

  18. Avian Interferons and Their Antiviral Effectors

    Science.gov (United States)

    Santhakumar, Diwakar; Rubbenstroth, Dennis; Martinez-Sobrido, Luis; Munir, Muhammad

    2017-01-01

    Interferon (IFN) responses, mediated by a myriad of IFN-stimulated genes (ISGs), are the most profound innate immune responses against viruses. Cumulatively, these IFN effectors establish a multilayered antiviral state to safeguard the host against invading viral pathogens. Considerable genetic and functional characterizations of mammalian IFNs and their effectors have been made, and our understanding on the avian IFNs has started to expand. Similar to mammalian counterparts, three types of IFNs have been genetically characterized in most avian species with available annotated genomes. Intriguingly, chickens are capable of mounting potent innate immune responses upon various stimuli in the absence of essential components of IFN pathways including retinoic acid-inducible gene I, IFN regulatory factor 3 (IRF3), and possibility IRF9. Understanding these unique properties of the chicken IFN system would propose valuable targets for the development of potential therapeutics for a broader range of viruses of both veterinary and zoonotic importance. This review outlines recent developments in the roles of avian IFNs and ISGs against viruses and highlights important areas of research toward our understanding of the antiviral functions of IFN effectors against viral infections in birds. PMID:28197148

  19. Avian Interferons and Their Antiviral Effectors.

    Science.gov (United States)

    Santhakumar, Diwakar; Rubbenstroth, Dennis; Martinez-Sobrido, Luis; Munir, Muhammad

    2017-01-01

    Interferon (IFN) responses, mediated by a myriad of IFN-stimulated genes (ISGs), are the most profound innate immune responses against viruses. Cumulatively, these IFN effectors establish a multilayered antiviral state to safeguard the host against invading viral pathogens. Considerable genetic and functional characterizations of mammalian IFNs and their effectors have been made, and our understanding on the avian IFNs has started to expand. Similar to mammalian counterparts, three types of IFNs have been genetically characterized in most avian species with available annotated genomes. Intriguingly, chickens are capable of mounting potent innate immune responses upon various stimuli in the absence of essential components of IFN pathways including retinoic acid-inducible gene I, IFN regulatory factor 3 (IRF3), and possibility IRF9. Understanding these unique properties of the chicken IFN system would propose valuable targets for the development of potential therapeutics for a broader range of viruses of both veterinary and zoonotic importance. This review outlines recent developments in the roles of avian IFNs and ISGs against viruses and highlights important areas of research toward our understanding of the antiviral functions of IFN effectors against viral infections in birds.

  20. Human influenza is more effective than avian influenza at antiviral suppression in airway cells.

    Science.gov (United States)

    Hsu, Alan Chen-Yu; Barr, Ian; Hansbro, Philip M; Wark, Peter A

    2011-06-01

    Airway epithelial cells are the initial site of infection with influenza viruses. The innate immune responses of airway epithelial cells to infection are important in limiting virus replication and spread. However, relatively little is known about the importance of this innate antiviral response to infection. Avian influenza viruses are a potential source of future pandemics; therefore, it is critical to examine the effectiveness of the host antiviral system to different influenza viruses. We used a human influenza (H3N2) and a low-pathogenic avian influenza (H11N9) to assess and compare the antiviral responses of Calu-3 cells. After infection, H3N2 replicated more effectively than the H11N9 in Calu-3 cells. This was not due to differential expression of sialic acid residues on Calu-3 cells, but was attributed to the interference of host antiviral responses by H3N2. H3N2 induced a delayed antiviral signaling and impaired type I and type III IFN induction compared with the H11N9. The gene encoding for nonstructural (NS) 1 protein was transfected into the bronchial epithelial cells (BECs), and the H3N2 NS1 induced a greater inhibition of antiviral responses compared with the H11N9 NS1. Although the low-pathogenic avian influenza virus was capable of infecting BECs, the human influenza virus replicated more effectively than avian influenza virus in BECs, and this was due to a differential ability of the two NS1 proteins to inhibit antiviral responses. This suggests that the subversion of human antiviral responses may be an important requirement for influenza viruses to adapt to the human host and cause disease.

  1. The FACT complex promotes avian leukosis virus DNA integration.

    Science.gov (United States)

    Winans, Shelby; Larue, Ross C; Abraham, Carly M; Shkriabai, Nikolozi; Skopp, Amelie; Winkler, Duane; Kvaratskhelia, Mamuka; Beemon, Karen L

    2017-01-25

    All retroviruses need to integrate a DNA copy of their genome into the host chromatin. Cellular proteins regulating and targeting lentiviral and gammaretroviral integration in infected cells have been discovered, but the factors that mediate alpharetroviral avian leukosis virus (ALV) integration are unknown. Here, we have identified the FACT protein complex, which consists of SSRP1 and Spt16, as a principal cellular binding partner of ALV integrase (IN). Biochemical experiments with purified recombinant proteins show that SSRP1 and Spt16 are able to individually bind ALV IN, but only the FACT complex effectively stimulates ALV integration activity in vitro Likewise, in infected cells, the FACT complex promotes ALV integration activity with proviral integration frequency varying directly with cellular expression levels of the FACT complex. An increase in 2-LTR circles in the depleted FACT complex cell line indicates that this complex regulates the ALV life cycle at the level of integration. This regulation is shown to be specific to ALV, as disruption of the FACT complex did not inhibit either lentiviral or gammaretroviral integration in infected cells.

  2. A novel high-throughput assay for islet respiration reveals uncoupling of rodent and human islets.

    Directory of Open Access Journals (Sweden)

    Jakob D Wikstrom

    Full Text Available BACKGROUND: The pancreatic beta cell is unique in its response to nutrient by increased fuel oxidation. Recent studies have demonstrated that oxygen consumption rate (OCR may be a valuable predictor of islet quality and long term nutrient responsiveness. To date, high-throughput and user-friendly assays for islet respiration are lacking. The aim of this study was to develop such an assay and to examine bioenergetic efficiency of rodent and human islets. METHODOLOGY/PRINCIPAL FINDINGS: The XF24 respirometer platform was adapted to islets by the development of a 24-well plate specifically designed to confine islets. The islet plate generated data with low inter-well variability and enabled stable measurement of oxygen consumption for hours. The F1F0 ATP synthase blocker oligomycin was used to assess uncoupling while rotenone together with myxothiazol/antimycin was used to measure the level of non-mitochondrial respiration. The use of oligomycin in islets was validated by reversing its effect in the presence of the uncoupler FCCP. Respiratory leak averaged to 59% and 49% of basal OCR in islets from C57Bl6/J and FVB/N mice, respectively. In comparison, respiratory leak of INS-1 cells and C2C12 myotubes was measured to 38% and 23% respectively. Islets from a cohort of human donors showed a respiratory leak of 38%, significantly lower than mouse islets. CONCLUSIONS/SIGNIFICANCE: The assay for islet respiration presented here provides a novel tool that can be used to study islet mitochondrial function in a relatively high-throughput manner. The data obtained in this study shows that rodent islets are less bioenergetically efficient than human islets as well as INS1 cells.

  3. Apoptosis-associated uncoupling of bone formation and resorption in osteomyelitis

    Science.gov (United States)

    Marriott, Ian

    2013-01-01

    The mechanisms underlying the destruction of bone tissue in osteomyelitis are only now being elucidated. While some of the tissue damage associated with osteomyelitis likely results from the direct actions of bacteria and infiltrating leukocytes, perhaps exacerbated by bacterial manipulation of leukocyte survival pathways, infection-induced bone loss predominantly results from an uncoupling of the activities of osteoblasts and osteoclasts. Bacteria or their products can directly increase osteoclast formation and activity, and the inflammatory milieu at sites of infection can further promote bone resorption. In addition, osteoclast activity is critically regulated by osteoblasts that can respond to bacterial pathogens and foster both inflammation and osteoclastogenesis. Importantly, bone loss during osteomyelitis is also brought about by a decline in new bone deposition due to decreased bone matrix synthesis and by increased rates of osteoblast apoptosis. Extracellular bacterial components may be sufficient to reduce osteoblast viability, but the causative agents of osteomyelitis are also capable of inducing continuous apoptosis of these cells by activating intrinsic and extrinsic cell death pathways to further uncouple bone formation and resorption. Interestingly, bacterial internalization appears to be required for maximal osteoblast apoptosis, and cytosolic inflammasome activation may act in concert with autocrine/paracrine death receptor-ligand signaling to induce cell death. The manipulation of apoptotic pathways in infected bone cells could be an attractive new means to limit inflammatory damage in osteomyelitis. However, the mechanism that is the most important in bacterium-induced bone loss has not yet been identified. Furthermore, it remains to be determined whether the host would be best served by preventing osteoblast cell death or by promoting apoptosis in infected cells. PMID:24392356

  4. Uncoupled embryonic and extra-embryonic tissues compromise blastocyst development after somatic cell nuclear transfer.

    Directory of Open Access Journals (Sweden)

    Séverine A Degrelle

    Full Text Available Somatic cell nuclear transfer (SCNT is the most efficient cell reprogramming technique available, especially when working with bovine species. Although SCNT blastocysts performed equally well or better than controls in the weeks following embryo transfer at Day 7, elongation and gastrulation defects were observed prior to implantation. To understand the developmental implications of embryonic/extra-embryonic interactions, the morphological and molecular features of elongating and gastrulating tissues were analysed. At Day 18, 30 SCNT conceptuses were compared to 20 controls (AI and IVP: 10 conceptuses each; one-half of the SCNT conceptuses appeared normal while the other half showed signs of atypical elongation and gastrulation. SCNT was also associated with a high incidence of discordance in embryonic and extra-embryonic patterns, as evidenced by morphological and molecular "uncoupling". Elongation appeared to be secondarily affected; only 3 of 30 conceptuses had abnormally elongated shapes and there were very few differences in gene expression when they were compared to the controls. However, some of these differences could be linked to defects in microvilli formation or extracellular matrix composition and could thus impact extra-embryonic functions. In contrast to elongation, gastrulation stages included embryonic defects that likely affected the hypoblast, the epiblast, or the early stages of their differentiation. When taking into account SCNT conceptus somatic origin, i.e. the reprogramming efficiency of each bovine ear fibroblast (Low: 0029, Med: 7711, High: 5538, we found that embryonic abnormalities or severe embryonic/extra-embryonic uncoupling were more tightly correlated to embryo loss at implantation than were elongation defects. Alternatively, extra-embryonic differences between SCNT and control conceptuses at Day 18 were related to molecular plasticity (high efficiency/high plasticity and subsequent pregnancy loss. Finally

  5. China makes an impressive breakthrough in avian influenza virus research - Discovering the "heart" of avian infl uenza virus.

    Science.gov (United States)

    Li, Y G; Wu, J F; Li, X

    2009-02-01

    achieved exciting results in providing a detailed understanding of the mechanisms of action of the RNA polymerase PA subunit, the "heart" of the avian influenza virus, at the atomic level. They hope to provide clues to potential avian influenza therapy targets and a new platform for new drug discovery (http://202.123.110.5/jrzg/2009-02/06/content_1222973.htm, available as of February 6, 2009). According to Liu et al., influenza viruses are enveloped, negatively stranded RNA viruses with a segmented genome (consisting of 8 RNA segments) that can encode 11 kinds of viral proteins. Among these proteins, the complex of influenza polymerase, consisting of PB1, PB2, and PA subunits, is regarded to be what gives life to influenza viruses because of its essential catalytic role in viral RNA replication and mRNA transcription in the nucleus of infected cells. Notwithstanding earlier virology studies on the influenza virus that elucidated the functions of PB1 and PB2, the exact function of PA is still not completely clear. The group resolved the crystal structure of the carboxyl-terminus of PA in complex with the aminoterminus of PB1 peptides for the first time. This structure mode provides details for the interactions of PA and PB1, as well as the binding sites of PA and RNA. Results of the research, entitled the "Crystal structure of the polymerase PA(c)-CPB1(N) complex from an avian influenza H5N1 virus," were published in the August 28th issue of the respected international scientific journal Nature (He X, Zhou J, Bartlam M, et al. Nature. 2008; 454:1123-1126). Further efforts by the group served to indicate the fine three-dimensional structure of the N-terminal of PA protein. They revealed that the PA subunit holds an endonuclease active site and that it, rather than the PB1 subunit as was previously, plays a critical role in the endonuclease activity of influenza virus polymerase. In addition, PA's characteristics of being highly conserved and having little mutations make it

  6. Molecular cloning and functional characterization of avian interleukin-19

    Science.gov (United States)

    The present study describes the cloning and functional characterization of avian interleukin (IL)-19, a cytokine that, in mammals, alters the balance of Th1 and Th2 cells in favor of the Th2 phenotype. The full-length avian IL-19 gene, located on chromosome 26, was amplified from LPS-stimulated chi...

  7. 9 CFR 113.326 - Avian Pox Vaccine.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Avian Pox Vaccine. 113.326 Section 113... Vaccines § 113.326 Avian Pox Vaccine. Fowl Pox Vaccine and Pigeon Pox Vaccine shall be prepared from virus... this section shall be used for preparing the production seed virus for vaccine production. All...

  8. Genetic differences between avian and human isolates of Candida dubliniensis.

    LENUS (Irish Health Repository)

    McManus, Brenda A

    2009-09-01

    When Candida dubliniensis isolates obtained from seabird excrement and from humans in Ireland were compared by using multilocus sequence typing, 13 of 14 avian isolates were genetically distinct from human isolates. The remaining avian isolate was indistinguishable from a human isolate, suggesting that transmission may occur between humans and birds.

  9. Comparative genomics reveals insights into avian genome evolution and adaptation

    DEFF Research Database (Denmark)

    Zhang, Guojie; Li, Cai; Li, Qiye

    2014-01-01

    Birds are the most species-rich class of tetrapod vertebrates and have wide relevance across many research fields. We explored bird macroevolution using full genomes from 48 avian species representing all major extant clades. The avian genome is principally characterized by its constrained size, ...

  10. Putative Novel Genotype of Avian Hepatitis E Virus, Hungary, 2010

    OpenAIRE

    Bányai, Krisztián; Tóth, Ádám György; Ivanics, Éva; Glávits, Róbert; Szentpáli-Gavallér, Katalin; Dán, Ádám

    2012-01-01

    To explore the genetic diversity of avian hepatitis E virus strains, we characterized the near-complete genome of a strain detected in 2010 in Hungary, uncovering moderate genome sequence similarity with reference strains. Public health implications related to consumption of eggs or meat contaminated by avian hepatitis E virus, or to poultry handling, require thorough investigation.

  11. Putative novel genotype of avian hepatitis E virus, Hungary, 2010.

    Science.gov (United States)

    Bányai, Krisztián; Tóth, Ádám György; Ivanics, Éva; Glávits, Róbert; Szentpáli-Gavallér, Katalin; Dán, Ádám

    2012-08-01

    To explore the genetic diversity of avian hepatitis E virus strains, we characterized the near-complete genome of a strain detected in 2010 in Hungary, uncovering moderate genome sequence similarity with reference strains. Public health implications related to consumption of eggs or meat contaminated by avian hepatitis E virus, or to poultry handling, require thorough investigation.

  12. China's Cool Handling of Avian Flu

    Institute of Scientific and Technical Information of China (English)

    LIWUZHOU

    2004-01-01

    ON January 27, 2004,the China National Avian Flu Reference Lab confirmed that in Dingdang Town, Long'an County,Guangxi Zhuang Autonomous Region a duck had died of the highly pathogenic H5N1 avian influenza. In contrast to the SARS epidemic last year, this occurrence has been handled coolly and efficiently by the Chinese government and people in general.

  13. Avian Influenza Viruses in Water Birds, Africa 1

    OpenAIRE

    Gaidet, Nicolas; Dodman, Tim; Caron, Alexandre; Balança, Gilles; Desvaux, Stephanie; Goutard, Flavie; Cattoli, Giovanni; Lamarque, François; Hagemeijer, Ward; Monicat, François

    2007-01-01

    We report the first large-scale surveillance of avian influenza viruses in water birds conducted in Africa. This study shows evidence of avian influenza viruses in wild birds, both Eurasian and Afro-tropical species, in several major wetlands of Africa.

  14. Poultry-handling Practices during Avian Influenza Outbreak, Thailand

    OpenAIRE

    Sonja J Olsen; Laosiritaworn, Yongjua; Pattanasin, Sarika; Prapasiri, Prabda; Scott F Dowell

    2005-01-01

    With poultry outbreaks of avian influenza H5N1 continuing in Thailand, preventing human infection remains a priority. We surveyed residents of rural Thailand regarding avian influenza knowledge, attitudes, and practices. Results suggest that public education campaigns have been effective in reaching those at greatest risk, although some high-risk behavior continues.

  15. Avian influenza in shorebirds: experimental infection of ruddy turnstones (Arenaria interpres) with avian influenza virus

    Science.gov (United States)

    Hall, Jeffrey S.; Krauss, Scott; Franson, J. Christian; TeSlaa, Joshua L.; Nashold, Sean W.; Stallknecht, David E.; Webby, Richard J.; Webster, Robert G.

    2013-01-01

    Background: Low pathogenic avian influenza viruses (LPAIV) have been reported in shorebirds, especially at Delaware Bay, USA, during spring migration. However, data on patterns of virus excretion, minimal infectious doses, and clinical outcome are lacking. The ruddy turnstone (Arenaria interpres) is the shorebird species with the highest prevalence of influenza virus at Delaware Bay. Objectives: The primary objective of this study was to experimentally assess the patterns of influenza virus excretion, minimal infectious doses, and clinical outcome in ruddy turnstones. Methods: We experimentally challenged ruddy turnstones using a common LPAIV shorebird isolate, an LPAIV waterfowl isolate, or a highly pathogenic H5N1 avian influenza virus. Cloacal and oral swabs and sera were analyzed from each bird. Results: Most ruddy turnstones had pre-existing antibodies to avian influenza virus, and many were infected at the time of capture. The infectious doses for each challenge virus were similar (103·6–104·16 EID50), regardless of exposure history. All infected birds excreted similar amounts of virus and showed no clinical signs of disease or mortality. Influenza A-specific antibodies remained detectable for at least 2 months after inoculation. Conclusions: These results provide a reference for interpretation of surveillance data, modeling, and predicting the risks of avian influenza transmission and movement in these important hosts.

  16. Exploring the avian gut microbiota: current trends and future directions.

    Science.gov (United States)

    Waite, David W; Taylor, Michael W

    2015-01-01

    Birds represent a diverse and evolutionarily successful lineage, occupying a wide range of niches throughout the world. Like all vertebrates, avians harbor diverse communities of microorganisms within their guts, which collectively fulfill crucial roles in providing the host with nutrition and protection from pathogens. Across the field of avian microbiology knowledge is extremely uneven, with several species accounting for an overwhelming majority of all microbiological investigations. These include agriculturally important birds, such as chickens and turkeys, as well as birds of evolutionary or conservation interest. In our previous study we attempted the first meta-analysis of the avian gut microbiota, using 16S rRNA gene sequences obtained from a range of publicly available data sets. We have now extended our analysis to explore the microbiology of several key species in detail, to consider the avian microbiota within the context of what is known about other vertebrates, and to identify key areas of interest in avian microbiology for future study.

  17. Hemagglutinin protein of avian influenza virus H5N1 in immune injury of peripheral immune organs and its possible mechanism%H5N1病毒HA蛋白对小鼠外周免疫器官的损伤及其机制

    Institute of Scientific and Technical Information of China (English)

    葛南海; 胥韦; 陈敏惠; 徐军

    2012-01-01

    This study aims to investigate the role of recombinant hemagglutinin protein (HA) of avian influenza virus (AVI) H5N1 in immune injury of peripheral immune organ spleen and the related cytokines secretion in mice. By using insect-baculovirus expression system and nickel affinity magnet beads, HA protein of AVI H5N1 was recombined and was used in mice. Seventy-two hours after HA protein instillation, the lungs and spleens were isolated form the mice. Pathological examination revealed spleen swelling, destruction of local structure of germinal center, vacuolar changes and a decrease in lymphocyte, while lung pathological examination showed diffuse alveolar damage, inflammatory cell infiltration in mice challenged by HA. The injury index of spleen cells after LPS stimulation in HA pretreatment was higher than that in those of PBS pretreatment. And under basal condition, the levels of IP-10, MCP-la and Rantes released from the spleen cells of mice in HA pretreatment were significantly higher than those from the mice of PBS pretreatment group (P< 0.05). Twenty-four hours after LPS stimulation, the levels of IFN-γ, IP-10, MCP-la, and Rantes released from the spleen cells of HA pre-treated mice was significantly higher than those from PBS pretreated mice (P< 0.05). In conclusion, the H5N1 virus HA membrane protein by intratracheal instillation elicits a strong pathological immune response in the host, resulting in immune injury of peripheral lymphoid organs and immune cells and acute lung injury.%目的 研究H5N1病毒HA蛋白作用于小鼠后对其外周免疫器官脾脏的损伤其相关细胞因子的分泌.方法 通过昆虫-杆状病毒表达系统表达H5N1病毒HA蛋白,并经镍亲和磁珠纯化,将重组的HA蛋白作用于小鼠,72 h后,取其脾脏,观察其镜下形态学变化;分离脾细胞并培养,检测其细胞因子的分泌.结果 脾组织病理表现为局部性弥漫性肺泡损伤,单核细胞侵润,脾脏肿胀,毛细血管充血,生发

  18. Development of a blocking ELISA for detection of antibodies against avian hepatitis E virus.

    Science.gov (United States)

    Liu, Baoyuan; Zhao, Qin; Sun, Yani; Wang, Xinjie; Zhao, Jinan; Du, Taofeng; Wang, Chengbao; Xiao, Shuqi; Mu, Yang; Zhang, Gaiping; Luo, Jianxun; Hsu, Walter H; Zhou, En-Min

    2014-08-01

    A blocking enzyme-linked immunosorbent assay (bELISA) was developed for the detection of immunoglobulin G antibodies against avian hepatitis E virus (HEV). In the bELISA, the coating antigen was a truncated protein containing C-terminal 268-amino acid region of ORF2 from an avian HEV strain isolated in China (CaHEV) and blocking antibody was a monoclonal antibody (mAb) 1H5 recognizing the epitope within amino acids 384-414 in the C-terminal 268-amino acid region. The concentration of blocking mAb 1H5 was determined as that yielded an OD450nm value of 1.0 for binding to the coating antigen and the antigen concentration and serum dilution were optimized using a checkerboard titration. A cut-off value of 20.7% at the mean percent inhibition plus 3 standard deviations was determined by testing 265 negative sera. The bELISA had a sensitivity of 98.3% by testing 116 positive sera from chickens infected experimentally with CaHEV and had no cross-reaction with other anti-avian virus antibodies. The compliance rates of the bELISA with indirect ELISA and Western blot were 83.7% and 93.3%, respectively, by testing 300 field chicken sera. These results suggested that the bELISA developed in this study can be used for detection of antibodies against avian HEV and showed high reproducibility compared with indirect ELISA and Western blot methods.

  19. Current genomic editing approaches in avian transgenesis.

    Science.gov (United States)

    Park, Tae Sub; Kang, Kyung Soo; Han, Jae Yong

    2013-09-01

    The chicken was domesticated from Red Jungle Fowl over 8000years ago and became one of the major food sources worldwide. At present, the poultry industry is one of the largest industrial animal stocks in the world, and its economic scale is expanding significantly with increasing consumption. Additionally, since Aristotle used chicken eggs as a model to provide remarkable insights into how life begins, chickens have been used as invaluable and powerful experimental materials for studying embryo development, immune systems, biomedical processes, and hormonal regulation. Combined with advancements in efficient transgenic technology, avian models have become even more important than would have been expected.

  20. USGS highly pathogenic avian influenza research strategy

    Science.gov (United States)

    Harris, M. Camille; Miles, A. Keith; Pearce, John M.; Prosser, Diann J.; Sleeman, Jonathan M.; Whalen, Mary E.

    2015-09-09

    Avian influenza viruses are naturally occurring in wild birds such as ducks, geese, swans, and gulls. These viruses generally do not cause illness in wild birds, however, when spread to poultry they can be highly pathogenic and cause illness and death in backyard and commercial farms. Outbreaks may cause devastating agricultural economic losses and some viral strains have the potential to infect people directly. Furthermore, the combination of avian influenza viruses with mammalian viruses can result in strains with the ability to transmit from person to person, possibly leading to viruses with pandemic potential. All known pandemic influenza viruses have had some genetic material of avian origin. Since 1996, a strain of highly pathogenic avian influenza (HPAI) virus, H5N1, has caused infection in wild birds, losses to poultry farms in Eurasia and North Africa, and led to the deaths of several hundred people. Spread of the H5N1 virus and other influenza strains from China was likely facilitated by migratory birds. In December 2014, HPAI was detected in poultry in Canada and migratory birds in the United States. Since then, HPAI viruses have spread to large parts of the United States and will likely continue to spread through migratory bird flyways and other mechanisms throughout North America. In the United States, HPAI viruses have severely affected the poultry industry with millions of domestic birds dead or culled. These strains of HPAI are not known to cause disease in humans; however, the Centers for Disease Control and Prevention (CDC) advise caution when in close contact with infected birds. Experts agree that HPAI strains currently circulating in wild birds of North America will likely persist for the next few years. This unprecedented situation presents risks to the poultry industry, natural resource management, and potentially human health. Scientific knowledge and decision support tools are urgently needed to understand factors affecting the persistence

  1. Avian artificial insemination and semen preservation

    Science.gov (United States)

    Gee, G.F.; Risser, Arthur C.; Todd, Frank S.

    1983-01-01

    Summary: Artificial insemination is a practical propagation tool that has been successful with a variety of birds. Cooperative, massage, and electroejaculation and modifications of these three basic methods of semen collection are described for a variety of birds. Semen color and consistency and sperm number, moti!ity, and morphology, as discussed, are useful indicators of semen quality, but the most reliable test of semen quality is the production of fertile eggs. Successful cryogenic preservation of avian semen with DMSO or glycerol as the cryoprotectant has been possible. Although the methods for preservation require special equipment, use of frozen semen requires only simple insemination supplies

  2. Phylogenetic analysis of Neuraminidase gene of avian influenza H5N1 subtype detected in Iran in 1390(2011

    Directory of Open Access Journals (Sweden)

    E Kord

    2013-09-01

    Background & aim: Among the various subtypes of avian influenza viruses, an H5N1 subtype virus with high pathogenicity is of great importance. The aim of this study was to determine the Phylogenetic analysis of neuraminidase gene of avian influenza virus subtype of the H5N1 in Iran in 1390. Methods: In this experimental study, two swab samples from chickens with suspected symptoms of avian influenza were tested by the World Health Organization recommendation. The neuraminidase gene of positive samples was amplified by RT-PCR technique. After sequencing the phylogenetic studies were analyzed using MEGA5 and Megalign. Results: Phylogenetic analysis showed that the virus belongs to the Clade 2.3.2.1 which is highly similar to the viruses that are identified in Mongolia in 2010. Also in the stem of this virus neuraminidase protein a number of 20 amino acid has been deleted at position 69-49. Conclusion: Due to findings of this study, it seems that the virus has entered by migratory wild birds with the origin of Mongolia. Key words: Influenza, Avian, Neuraminidase

  3. Host- and Strain-Specific Regulation of Influenza Virus Polymerase Activity by Interacting Cellular Proteins

    NARCIS (Netherlands)

    Bortz, Eric; Westera, Liset; Maamary, Jad; Steel, John; Albrecht, Randy A.; Manicassamy, Balaji; Chase, Geoffrey; Martinez-Sobrido, Luis; Schwemmle, Martin; Garcia-Sastre, Adolfo

    2011-01-01

    Highly pathogenic avian influenza A (HPAI) viruses of the H5N1 subtype have recently emerged from avian zoonotic reservoirs to cause fatal human disease. Adaptation of HPAI virus RNA-dependent RNA polymerase (PB1, PB2, and PA proteins) and nucleoprotein (NP) to interactions with mammalian host prote

  4. Site-directed mutation of arginine 282 to glutamate uncouples the movement of peptides and protons by the rabbit proton-peptide cotransporter PepT1.

    Science.gov (United States)

    Meredith, David

    2004-04-16

    A conserved positive residue in the seventh transmembrane domain of the mammalian proton-coupled di- and tripeptide transporter PepT1 has been shown by site-directed mutagenesis to be a key residue for protein function. Substitution of arginine 282 with a glutamate residue (R282E-PepT1) gave a protein at the plasma membrane of Xenopus laevis oocytes that was able to transport the non-hydrolyzable dipeptide [3H]d-Phe-l-Gln, although unlike the wild type, the rate of transport by R282E-PepT1 was independent of the extracellular pH level, and the substrate could not be accumulated above equilibrium. The binding affinity of the mutant transport protein was unchanged from the wild type. Thus, R282E-Pept1 appears to have been changed from a proton-driven to a facilitated transporter for peptides. In addition, peptide transport by R282E-PepT1 still induced depolarization as measured by microelectrode recordings of membrane potential. A more detailed study by two-electrode voltage clamping revealed that R282E-PepT1 behaved as a peptide-gated non-selective cation channel with the ion selectivity series lithium > sodium > N-methyl-d-glucamine at pH 7.4. There was also a proton conductance (comparing pH 7.4 and 8.4), and at pH 5.5 the predominant conductance was for potassium ions. Therefore, it can be concluded that changing arginine 282 to a glutamate not only uncouples the cotransport of protons and peptides of the wild-type PepT1 but also creates a peptide-gated cation channel in the protein.

  5. AN EPIZOOTIC OF EMERGING NOVEL AVIAN POX IN CARRION CROWS (CORVUS CORONE) AND LARGE-BILLED CROWS (CORVUS MACRORHYNCHOS) IN JAPAN.

    Science.gov (United States)

    Fukui, Daisuke; Nakamura, Makiko; Yamaguchi, Tsuyoshi; Takenaka, Makiko; Murakami, Mami; Yanai, Tokuma; Fukushi, Hideto; Yanagida, Kazumi; Bando, Gen; Matsuno, Keita; Nagano, Masashi; Tsubota, Toshio

    2016-04-28

    In 2006-10, an epizootic of emerging avian pox occurred in Carrion Crows ( Corvus corone ) and Large-billed Crows ( Corvus macrorhynchos ), leading to mortality of juvenile crows in Hokkaido, the northernmost island of Japan. We diagnosed 27 crows with proliferative skin lesions (19 carcasses and eight biopsied cases [one in zoo captivity]) as avian pox clinically, histopathologically by detection of Avipoxvirus-specific 4b core protein (P4b) gene, and epidemiologically. The fatal cases demonstrated intensively severe infection and aggressive lesions with secondary bacterial infection. Since the first identification of avian pox in Sapporo, Japan, in 2006, the frequency of mortality events has increased, peaking in 2007-08. Mortalities have subsequently occurred in other areas, suggesting disease expansion. In Sapporo, prevalence of avian pox evaluated by field censuses during 2007-12 was 17.6% (6.6-27.2%), peaked during 2007-08 and 2008-09, and then decreased. All diseased crows were juveniles, except for one adult. The number of crows assembling in the winter roosts had been stable for >10 yr; however, it declined in 2007-08, decreased by about 50% in 2008-09, and recovered to the previous level in 2009-10, correlated with the avian pox outbreak. Thus, avian pox probably contributed to the unusual crow population decline. All P4b sequences detected in six specimens in Sapporo were identical and different from any previously reported sequences. The sequence detected in the zoo-kept crow was distinct from any reported clades, and interspecies transmission was suspected. This report demonstrates an emerging novel avian pox in the Japanese avifauna and in global populations of Carrion Crows and Large-billed Crows. Longitudinal monitoring is needed to evaluate its impact on the crow population.

  6. Thiamine Deficiency--Induced Partial Necrosis and Mitochondrial Uncoupling in Neuroblastoma Cells Are Rapidly Reversed by Addition of Thiamine

    OpenAIRE

    Bettendorff, Lucien; Sluse, Francis; Goessens, Guy; Wins, Pierre; Grisar, Thierry

    1995-01-01

    Culture of neuroblastoma cells in a medium of low-thiamine concentration (6 nM) and in the presence of the transport inhibitor amprolium leads to the appearance of overt signs of necrosis; i.e., the chromatin condenses in dark patches, the oxygen consumption decreases, mitochondria are uncoupled, and their cristae are disorganized. Glutamate formed from glutamine is no longer oxidized and accumulates, suggesting that the thiamine diphosphate-dependent alpha-ketoglutarate dehydrogenase activit...

  7. Extracellular signal-regulated kinases control expression of G protein-coupled receptor kinase 2 (GRK2)

    DEFF Research Database (Denmark)

    Theilade, Juliane; Lerche Hansen, Jakob; Haunsø, Stig;

    2002-01-01

    G protein-coupled receptor kinase 2 (GRK2) phosphorylates G protein-coupled receptors resulting in uncoupling from G proteins. Receptors modulate GRK2 expression, however the mechanistic basis for this effect is largely unknown. Here we report a novel mechanism by which receptors use...

  8. Avian-like breathing mechanics in maniraptoran dinosaurs.

    Science.gov (United States)

    Codd, Jonathan R; Manning, Phillip L; Norell, Mark A; Perry, Steven F

    2008-01-22

    In 1868 Thomas Huxley first proposed that dinosaurs were the direct ancestors of birds and subsequent analyses have identified a suite of 'avian' characteristics in theropod dinosaurs. Ossified uncinate processes are found in most species of extant birds and also occur in extinct non-avian maniraptoran dinosaurs. Their presence in these dinosaurs represents another morphological character linking them to Aves, and further supports the presence of an avian-like air-sac respiratory system in theropod dinosaurs, prior to the evolution of flight. Here we report a phylogenetic analysis of the presence of uncinate processes in Aves and non-avian maniraptoran dinosaurs indicating that these were homologous structures. Furthermore, recent work on Canada geese has demonstrated that uncinate processes are integral to the mechanics of avian ventilation, facilitating both inspiration and expiration. In extant birds, uncinate processes function to increase the mechanical advantage for movements of the ribs and sternum during respiration. Our study presents a mechanism whereby uncinate processes, in conjunction with lateral and ventral movements of the sternum and gastral basket, affected avian-like breathing mechanics in extinct non-avian maniraptoran dinosaurs.

  9. Spatial uncoupling of biodegradation, soil respiration, and PAH concentration in a creosote contaminated soil

    Energy Technology Data Exchange (ETDEWEB)

    Bengtsson, Goeran, E-mail: goran.bengtsson@ekol.lu.s [Lund University, Department of Ecology, Soelvegatan 37, SE-223 62 Lund (Sweden); Toerneman, Niklas [Lund University, Department of Ecology, Soelvegatan 37, SE-223 62 Lund (Sweden); Yang Xiuhong [Lund University, Department of Ecology, Soelvegatan 37, SE-223 62 Lund (Sweden); Department of Environmental Science, School of Environmental Science and Engineering, Sun Yat-sen University, Guangzhou 510275 (China)

    2010-09-15

    Hotspots and coldspots of concentration and biodegradation of polycyclic aromatic hydrocarbons (PAHs) marginally overlapped at the 0.5-100 m scale in a creosote contaminated soil in southern Sweden, suggesting that concentration and biodegradation had little spatial co-variation. Biodegradation was substantial and its spatial variability considerable and highly irregular, but it had no spatial autocorrelation. The soil concentration of PAHs explained only 20-30% of the variance of their biodegradation. Soil respiration was spatially autocorrelated. The spatial uncoupling between biodegradation and soil respiration seemed to be governed by the aging of PAHs in the soil, since biodegradation of added {sup 13}C phenanthrene covaried with both soil respiration and microbial biomass. The latter two were also correlated with high concentrations of phospholipid fatty acids (PLFAs) that are common in gram-negative bacteria. However, several of the hotspots of biodegradation coincided with hotspots for the distribution of a PLFA indicative of fungal biomass. - Hotspots of PAH biodegradation in a creosote contaminated soil do not coincide with hotspots of PAH concentration, microbial biomass and respiration.

  10. Growth cessation uncouples isotopic signals in leaves and tree rings of drought-exposed oak trees.

    Science.gov (United States)

    Pflug, Ellen E; Siegwolf, R; Buchmann, N; Dobbertin, M; Kuster, T M; Günthardt-Goerg, M S; Arend, M

    2015-10-01

    An increase in temperature along with a decrease in summer precipitation in Central Europe will result in an increased frequency of drought events and gradually lead to a change in species composition in forest ecosystems. In the present study, young oaks (Quercus robur L. and Quercus petraea (Matt.) Liebl.) were transplanted into large mesocosms and exposed for 3 years to experimental warming and a drought treatment with yearly increasing intensities. Carbon and oxygen isotopic (δ(13)C and δ(18)O) patterns were analysed in leaf tissue and tree-ring cellulose and linked to leaf physiological measures and tree-ring growth. Warming had no effect on the isotopic patterns in leaves and tree rings, while drought increased δ(18)O and δ(13)C. Under severe drought, an unexpected isotopic pattern, with a decrease in δ(18)O, was observed in tree rings but not in leaves. This decrease in δ(18)O could not be explained by concurrent physiological analyses and is not supported by current physiological knowledge. Analysis of intra-annual tree-ring growth revealed a drought-induced growth cessation that interfered with the record of isotopic signals imprinted on recently formed leaf carbohydrates. This missing record indicates isotopic uncoupling of leaves and tree rings, which may have serious implications for the interpretation of tree-ring isotopes, particularly from trees that experienced growth-limiting stresses.

  11. Mitochondrial uncoupling and the reprogramming of intermediary metabolism in leukemia cells

    Directory of Open Access Journals (Sweden)

    Juliana eVélez

    2013-04-01

    Full Text Available Nearly 60 years ago Otto Warburg proposed, in a seminal publication, that an irreparable defect in the oxidative capacity of normal cells supported the switch to glycolysis for energy generation and the appearance of the malignant phenotype (Warburg, 1956. Curiously, this phenotype was also observed by Warburg in embryonic tissues, and recent research demonstrated that normal stem cells may indeed rely on aerobic glycolysis – fermenting pyruvate to lactate in the presence of ample oxygen - rather than on the complete oxidation of pyruvate in the Krebs cycle - to generate cellular energy (Folmes et al., 2012. However, it remains to be determined whether this phenotype is causative for neoplastic development, or rather the result of malignant transformation. In addition, in light of mounting evidence demonstrating that cancer cells can carry out electron transport and oxidative phosphorylation, although in some cases predominantly using electrons from non-glucose carbon sources (Bloch-Frankenthal et al., 1965, Warburg´s hypothesis needs to be revisited. Lastly, recent evidence suggests that the leukemia bone marrow microenvironment promotes the Warburg phenotype adding another layer of complexity to the study of metabolism in hematological malignancies. In this review we will discuss some of the evidence for alterations in the intermediary metabolism of leukemia cells and present evidence for a concept put forth decades ago by lipid biochemist Feodor Lynen, and acknowledged by Warburg himself, that cancer cell mitochondria uncouple ATP synthesis from electron transport and therefore depend on glycolysis to meet their energy demands (Lynen, 1951;Warburg, 1956.

  12. Temporal Uncoupling between Energy Acquisition and Allocation to Reproduction in a Herbivorous-Detritivorous Fish

    Science.gov (United States)

    Villamarín, Francisco; Magnusson, William E.; Jardine, Timothy D.; Valdez, Dominic; Woods, Ryan; Bunn, Stuart E.

    2016-01-01

    Although considerable knowledge has been gathered regarding the role of fish in cycling and translocation of nutrients across ecosystem boundaries, little information is available on how the energy obtained from different ecosystems is temporally allocated in fish bodies. Although in theory, limitations on energy budgets promote the existence of a trade-off between energy allocated to reproduction and somatic growth, this trade-off has rarely been found under natural conditions. Combining information on RNA:DNA ratios and carbon and nitrogen stable-isotope analyses we were able to achieve novel insights into the reproductive allocation of diamond mullet (Liza alata), a catadromous, widely distributed herbivorous-detritivorous fish. Although diamond mullet were in better condition during the wet season, most reproductive allocation occurred during the dry season when resources are limited and fish have poorer body condition. We found a strong trade-off between reproductive and somatic investment. Values of δ13C from reproductive and somatic tissues were correlated, probably because δ13C in food resources between dry and wet seasons do not differ markedly. On the other hand, data for δ15N showed that gonads are more correlated to muscle, a slow turnover tissue, suggesting long term synthesis of reproductive tissues. In combination, these lines of evidence suggest that L. alata is a capital breeder which shows temporal uncoupling of resource ingestion, energy storage and later allocation to reproduction. PMID:26938216

  13. A quinoxaline urea analog uncouples inflammatory and pro-survival functions of IKKβ.

    Science.gov (United States)

    Maroni, Dulce; Rana, Sandeep; Mukhopadhyay, Chandrani; Natarajan, Amarnath; Naramura, Mayumi

    2015-12-01

    Activation of the NF-κB pathway is causally linked to initiation and progression of diverse cancers. Therefore, IKKβ, the key regulatory kinase of the canonical NF-κB pathway, should be a logical target for cancer treatment. However, existing IKKβ inhibitors are known to induce paradoxical immune activation, which limits their clinical usefulness. Recently, we identified a quinoxaline urea analog 13-197 as a novel IKKβ inhibitor that delays tumor growth without significant adverse effects in xenograft tumor models. In the present study, we found that 13-197 had little effect on LPS-induced NF-κB target gene induction by primary mouse macrophages while maintaining considerable anti-proliferative activities. These characteristics may explain absence of inflammatory side effects in animals treated with 13-197. Our data also demonstrate that the inflammation and proliferation-related functions of IKKβ can be uncoupled, and highlight the utility of 13-197 to dissect these downstream pathways.

  14. TGF-beta Sma/Mab signaling mutations uncouple reproductive aging from somatic aging.

    Directory of Open Access Journals (Sweden)

    Shijing Luo

    2009-12-01

    Full Text Available Female reproductive cessation is one of the earliest age-related declines humans experience, occurring in mid-adulthood. Similarly, Caenorhabditis elegans' reproductive span is short relative to its total life span, with reproduction ceasing about a third into its 15-20 day adulthood. All of the known mutations and treatments that extend C. elegans' reproductive period also regulate longevity, suggesting that reproductive span is normally linked to life span. C. elegans has two canonical TGF-beta signaling pathways. We recently found that the TGF-beta Dauer pathway regulates longevity through the Insulin/IGF-1 Signaling (IIS pathway; here we show that this pathway has a moderate effect on reproductive span. By contrast, TGF-beta Sma/Mab signaling mutants exhibit a substantially extended reproductive period, more than doubling reproductive span in some cases. Sma/Mab mutations extend reproductive span disproportionately to life span and act independently of known regulators of somatic aging, such as Insulin/IGF-1 Signaling and Dietary Restriction. This is the first discovery of a pathway that regulates reproductive span independently of longevity and the first identification of the TGF-beta Sma/Mab pathway as a regulator of reproductive aging. Our results suggest that longevity and reproductive span regulation can be uncoupled, although they appear to normally be linked through regulatory pathways.

  15. Temporal Uncoupling between Energy Acquisition and Allocation to Reproduction in a Herbivorous-Detritivorous Fish.

    Directory of Open Access Journals (Sweden)

    Francisco Villamarín

    Full Text Available Although considerable knowledge has been gathered regarding the role of fish in cycling and translocation of nutrients across ecosystem boundaries, little information is available on how the energy obtained from different ecosystems is temporally allocated in fish bodies. Although in theory, limitations on energy budgets promote the existence of a trade-off between energy allocated to reproduction and somatic growth, this trade-off has rarely been found under natural conditions. Combining information on RNA:DNA ratios and carbon and nitrogen stable-isotope analyses we were able to achieve novel insights into the reproductive allocation of diamond mullet (Liza alata, a catadromous, widely distributed herbivorous-detritivorous fish. Although diamond mullet were in better condition during the wet season, most reproductive allocation occurred during the dry season when resources are limited and fish have poorer body condition. We found a strong trade-off between reproductive and somatic investment. Values of δ13C from reproductive and somatic tissues were correlated, probably because δ13C in food resources between dry and wet seasons do not differ markedly. On the other hand, data for δ15N showed that gonads are more correlated to muscle, a slow turnover tissue, suggesting long term synthesis of reproductive tissues. In combination, these lines of evidence suggest that L. alata is a capital breeder which shows temporal uncoupling of resource ingestion, energy storage and later allocation to reproduction.

  16. Mechanisms of excitation-contraction uncoupling relevant to activity-induced muscle fatigue.

    Science.gov (United States)

    Lamb, Graham D

    2009-06-01

    If the free [Ca2+] in the cytoplasm of a skeletal muscle fiber is raised substantially for a period of seconds to minutes or to high levels just briefly, it leads to disruption of the normal excitation-contraction (E-C) coupling process and a consequent long-lasting decrease in force production. It appears that the disruption to the coupling occurs at the triad junction, where the voltage-sensor molecules (dihydropyridine receptors) normally interact with and open the Ca2+ release channels (ryanodine receptors) in the adjacent sarcoplasmic reticulum (SR). This disruption results in inadequate release of SR Ca2+ upon stimulation. Such E-C uncoupling may underlie the long-duration low-frequency fatigue that can occur after various types of exercise, as well as possibly being a contributing factor to the muscle weakness in certain muscle diseases. The process or processes causing the disruption of the coupling between the voltage sensors and the release channels is not known with certainty, but might be associated with structural changes at the triad junction, possibly caused by activation of the Ca2+-dependent protease, micro-calpain.

  17. Uncoupling charge movement from channel opening in voltage-gated potassium channels by ruthenium complexes.

    Science.gov (United States)

    Jara-Oseguera, Andrés; Ishida, Itzel G; Rangel-Yescas, Gisela E; Espinosa-Jalapa, Noel; Pérez-Guzmán, José A; Elías-Viñas, David; Le Lagadec, Ronan; Rosenbaum, Tamara; Islas, León D

    2011-05-06

    The Kv2.1 channel generates a delayed-rectifier current in neurons and is responsible for modulation of neuronal spike frequency and membrane repolarization in pancreatic β-cells and cardiomyocytes. As with other tetrameric voltage-activated K(+)-channels, it has been proposed that each of the four Kv2.1 voltage-sensing domains activates independently upon depolarization, leading to a final concerted transition that causes channel opening. The mechanism by which voltage-sensor activation is coupled to the gating of the pore is still not understood. Here we show that the carbon-monoxide releasing molecule 2 (CORM-2) is an allosteric inhibitor of the Kv2.1 channel and that its inhibitory properties derive from the CORM-2 ability to largely reduce the voltage dependence of the opening transition, uncoupling voltage-sensor activation from the concerted opening transition. We additionally demonstrate that CORM-2 modulates Shaker K(+)-channels in a similar manner. Our data suggest that the mechanism of inhibition by CORM-2 may be common to voltage-activated channels and that this compound should be a useful tool for understanding the mechanisms of electromechanical coupling.

  18. Weight loss by Ppc-1, a novel small molecule mitochondrial uncoupler derived from slime mold.

    Directory of Open Access Journals (Sweden)

    Toshiyuki Suzuki

    Full Text Available Mitochondria play a key role in diverse processes including ATP synthesis and apoptosis. Mitochondrial function can be studied using inhibitors of respiration, and new agents are valuable for discovering novel mechanisms involved in mitochondrial regulation. Here, we screened small molecules derived from slime molds and other microorganisms for their effects on mitochondrial oxygen consumption. We identified Ppc-1 as a novel molecule which stimulates oxygen consumption without adverse effects on ATP production. The kinetic behavior of Ppc-1 suggests its function as a mitochondrial uncoupler. Serial administration of Ppc-1 into mice suppressed weight gain with no abnormal effects on liver or kidney tissues, and no evidence of tumor formation. Serum fatty acid levels were significantly elevated in mice treated with Ppc-1, while body fat content remained low. After a single administration, Ppc-1 distributes into various tissues of individual animals at low levels. Ppc-1 stimulates adipocytes in culture to release fatty acids, which might explain the elevated serum fatty acids in Ppc-1-treated mice. The results suggest that Ppc-1 is a unique mitochondrial regulator which will be a valuable tool for mitochondrial research as well as the development of new drugs to treat obesity.

  19. Mitochondrial uncoupling does not decrease reactive oxygen species production after ischemia-reperfusion.

    Science.gov (United States)

    Quarrie, Ricardo; Lee, Daniel S; Reyes, Levy; Erdahl, Warren; Pfeiffer, Douglas R; Zweier, Jay L; Crestanello, Juan A

    2014-10-01

    Cardiac ischemia-reperfusion (IR) leads to myocardial dysfunction by increasing production of reactive oxygen species (ROS). Mitochondrial H(+) leak decreases ROS formation; it has been postulated that increasing H(+) leak may be a mechanism of decreasing ROS production after IR. Ischemic preconditioning (IPC) decreases ROS formation after IR, but the mechanism is unknown. We hypothesize that pharmacologically increasing mitochondrial H(+) leak would decrease ROS production after IR. We further hypothesize that IPC would be associated with an increase in the rate of H(+) leak. Isolated male Sprague-Dawley rat hearts were subjected to either control or IPC. Mitochondria were isolated at end equilibration, end ischemia, and end reperfusion. Mitochondrial membrane potential (mΔΨ) was measured using a tetraphenylphosphonium electrode. Mitochondrial uncoupling was achieved by adding increasing concentrations of FCCP. Mitochondrial ROS production was measured by fluorometry using Amplex-Red. Pyridine dinucleotide levels were measured using HPLC. Before IR, increasing H(+) leak decreased mitochondrial ROS production. After IR, ROS production was not affected by increasing H(+) leak. H(+) leak increased at end ischemia in control mitochondria. IPC mitochondria showed no change in the rate of H(+) leak throughout IR. NADPH levels decreased after IR in both IPC and control mitochondria while NADH increased. Pharmacologically, increasing H(+) leak is not a method of decreasing ROS production after IR. Replenishing the NADPH pool may be a means of scavenging the excess ROS thereby attenuating oxidative damage after IR.

  20. Is there a layer deep in the Earth that uncouples heat from mechanical work?

    Directory of Open Access Journals (Sweden)

    S. J. Burns

    2014-02-01

    Full Text Available The thermal expansion coefficient is presented as the coupling between heat energy and mechanical work. It is shown that when heat and work are uncoupled then very unusual material properties occurs: for example, acoustic p waves are not damped and heat is not generated from mechanical motion. It is found that at pressures defined by the bulk modulus divided by the Anderson–Grüneisen parameter, then the thermal expansion coefficient approaches zero in linear-elastic models. Very large pressures always reduce thermal expansion coefficients; the importance of a very small or even negative thermal expansion coefficient is discussed in relation to physical processes deep in the core and mantle of Earth. Models of the thermal expansion coefficients based on interatomic potentials which are always relegated to isometric conditions preclude any changes in volume due to temperature changes. However, it is known that the pressures in the Earth are large enough to effectively reduce thermal expansion coefficients to near zero which decouples heat from mechanical work.

  1. Pharmacologically-induced neurovascular uncoupling is associated with cognitive impairment in mice.

    Science.gov (United States)

    Tarantini, Stefano; Hertelendy, Peter; Tucsek, Zsuzsanna; Valcarcel-Ares, M Noa; Smith, Nataliya; Menyhart, Akos; Farkas, Eszter; Hodges, Erik L; Towner, Rheal; Deak, Ferenc; Sonntag, William E; Csiszar, Anna; Ungvari, Zoltan; Toth, Peter

    2015-11-01

    There is increasing evidence that vascular risk factors, including aging, hypertension, diabetes mellitus, and obesity, promote cognitive impairment; however, the underlying mechanisms remain obscure. Cerebral blood flow (CBF) is adjusted to neuronal activity via neurovascular coupling (NVC) and this mechanism is known to be impaired in the aforementioned pathophysiologic conditions. To establish a direct relationship between impaired NVC and cognitive decline, we induced neurovascular uncoupling pharmacologically in mice by inhibiting the synthesis of vasodilator mediators involved in NVC. Treatment of mice with the epoxygenase inhibitor N-(methylsulfonyl)-2-(2-propynyloxy)-benzenehexanamide (MSPPOH), the NO synthase inhibitor l-NG-Nitroarginine methyl ester (L-NAME), and the COX inhibitor indomethacin decreased NVC by over 60% mimicking the aging phenotype, which was associated with significantly impaired spatial working memory (Y-maze), recognition memory (Novel object recognition), and impairment in motor coordination (Rotarod). Blood pressure (tail cuff) and basal cerebral perfusion (arterial spin labeling perfusion MRI) were unaffected. Thus, selective experimental disruption of NVC is associated with significant impairment of cognitive and sensorimotor function, recapitulating neurologic symptoms and signs observed in brain aging and pathophysiologic conditions associated with accelerated cerebromicrovascular aging.

  2. Temporal Uncoupling between Energy Acquisition and Allocation to Reproduction in a Herbivorous-Detritivorous Fish.

    Science.gov (United States)

    Villamarín, Francisco; Magnusson, William E; Jardine, Timothy D; Valdez, Dominic; Woods, Ryan; Bunn, Stuart E

    2016-01-01

    Although considerable knowledge has been gathered regarding the role of fish in cycling and translocation of nutrients across ecosystem boundaries, little information is available on how the energy obtained from different ecosystems is temporally allocated in fish bodies. Although in theory, limitations on energy budgets promote the existence of a trade-off between energy allocated to reproduction and somatic growth, this trade-off has rarely been found under natural conditions. Combining information on RNA:DNA ratios and carbon and nitrogen stable-isotope analyses we were able to achieve novel insights into the reproductive allocation of diamond mullet (Liza alata), a catadromous, widely distributed herbivorous-detritivorous fish. Although diamond mullet were in better condition during the wet season, most reproductive allocation occurred during the dry season when resources are limited and fish have poorer body condition. We found a strong trade-off between reproductive and somatic investment. Values of δ13C from reproductive and somatic tissues were correlated, probably because δ13C in food resources between dry and wet seasons do not differ markedly. On the other hand, data for δ15N showed that gonads are more correlated to muscle, a slow turnover tissue, suggesting long term synthesis of reproductive tissues. In combination, these lines of evidence suggest that L. alata is a capital breeder which shows temporal uncoupling of resource ingestion, energy storage and later allocation to reproduction.

  3. Avian colibacillosis: still many black holes.

    Science.gov (United States)

    Guabiraba, Rodrigo; Schouler, Catherine

    2015-08-01

    Avian pathogenic Escherichia coli (APEC) strains cause severe respiratory and systemic diseases, threatening food security and avian welfare worldwide. Intensification of poultry production and the quick expansion of free-range production systems will increase the incidence of colibacillosis through greater exposure of birds to pathogens and stress. Therapy is mainly based on antibiotherapy and current vaccines have poor efficacy. Serotyping remains the most frequently used diagnostic method, only allowing the identification of a limited number of APEC strains. Several studies have demonstrated that the most common virulence factors studied in APEC are all rarely present in the same isolate, showing that APEC strains constitute a heterogeneous group. Different isolates may harbor different associations of virulence factors, each one able to induce colibacillosis. Despite its economical relevance, pathogenesis of colibacillosis is poorly understood. Our knowledge on the host response to APEC is based on very descriptive studies, mostly restricted to bacteriological and histopathological analysis of infected organs such as lungs. Furthermore, only a small number of APEC isolates have been used in experimental studies. In the present review, we discuss current knowledge on APEC diversity and virulence, including host response to infection and the associated inflammatory response with a focus on pulmonary colibacillosis.

  4. Collapsing avian community on a Hawaiian island

    Science.gov (United States)

    Paxton, Eben; Camp, Richard J.; Gorresen, P. Marcos; Crampton, Lisa H.; Leonard, David L.; VanderWerf, Eric

    2016-01-01

    The viability of many species has been jeopardized by numerous negative factors over the centuries, but climate change is predicted to accelerate and increase the pressure of many of these threats, leading to extinctions. The Hawaiian honeycreepers, famous for their spectacular adaptive radiation, are predicted to experience negative responses to climate change, given their susceptibility to introduced disease, the strong linkage of disease distribution to climatic conditions, and their current distribution. We document the rapid collapse of the native avifauna on the island of Kaua‘i that corresponds to changes in climate and disease prevalence. Although multiple factors may be pressuring the community, we suggest that a tipping point has been crossed in which temperatures in forest habitats at high elevations have reached a threshold that facilitates the development of avian malaria and its vector throughout these species’ ranges. Continued incursion of invasive weeds and non-native avian competitors may be facilitated by climate change and could also contribute to declines. If current rates of decline continue, we predict multiple extinctions in the coming decades. Kaua‘i represents an early warning for the forest bird communities on the Maui and Hawai‘i islands, as well as other species around the world that are trapped within a climatic space that is rapidly disappearing.

  5. Uncoupling of sarcoplasmic reticulum Ca²⁺-ATPase by N-arachidonoyl dopamine. Members of the endocannabinoid family as thermogenic drugs

    DEFF Research Database (Denmark)

    Mahmmoud, Yasser Ahmed; Gaster, Michel

    2013-01-01

    agents. EXPERIMENTAL APPROACH: Using isolated SR vesicles from rabbits, we have screened for endogenous compounds that uncouple SERCA. We have also studied their ability to deplete cytoplasmic ATP from human skeletal muscle cells in culture. KEY RESULTS: Studies on SR vesicles showed that the endogenous...... post-mitochondrial strategy for reduction of cellular ATP levels. In addition, signalling networks leading to SERCA uncoupling can be explored to study the importance of this ion pump in pathophysiological conditions related to metabolism....

  6. Complex formation between the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) and valinomycin in the presence of potassium.

    Science.gov (United States)

    O'Brien, T A; Nieva-Gomez, D; Gennis, R B

    1978-03-25

    Spectroscopic evidence is presented which indicates that the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) and the peptide antibiotic valinomycin form a complex in the presence of potassium. Complex formation has been observed both in aqueous and nonaqueous media. Several techniques have been used to indicate the existence of a complex in aqueous solution. In the presence of valinomycin and K+, the absorption spectrum of FCCP is significantly perturbed, and there is also a large induced circular dichroism signal. In addition, the previously characterized complex which forms between valinomycin, K+, and the fluorescent probe 8-anilino-1-naphthalene-sulfonate (ANS) in aqueous solution is apparently disrupted by the addition of FCCP. The result is an effective quenching of the fluorescence due to the bound probe as it is displaced from the valinomycin.K+ by the uncoupler. In a nonpolar solvent, the absorption spectrum of FCCP is also perturbed by valinomycin in the presence of K+, again indicating the formation of a complex. These data point to the importance of considering the role of valinomycin.K+.uncoupler complex in interpreting physiological or ion transport data in which these substances have been used together.

  7. A recombinant avian leukosis virus subgroup j for directly monitoring viral infection and the selection of neutralizing antibodies.

    Directory of Open Access Journals (Sweden)

    Qi Wang

    Full Text Available Avian leukosis virus subgroup J (ALV-J has induced serious clinical outbreaks and has become a serious infectious disease of chickens in China. We describe here the creation of a recombinant ALV-J tagged with the enhanced green fluorescent protein (named rHPRS-103EGFP. We successfully utilize the rHPRS-103EGFP to visualize viral infection and for development of a simplified serum-neutralization test.

  8. A recombinant avian leukosis virus subgroup j for directly monitoring viral infection and the selection of neutralizing antibodies.

    Science.gov (United States)

    Wang, Qi; Li, Xiaofei; Ji, Xiaolin; Wang, Jingfei; Shen, Nan; Gao, Yulong; Qi, Xiaole; Wang, Yongqiang; Gao, Honglei; Zhang, Shide; Wang, Xiaomei

    2014-01-01

    Avian leukosis virus subgroup J (ALV-J) has induced serious clinical outbreaks and has become a serious infectious disease of chickens in China. We describe here the creation of a recombinant ALV-J tagged with the enhanced green fluorescent protein (named rHPRS-103EGFP). We successfully utilize the rHPRS-103EGFP to visualize viral infection and for development of a simplified serum-neutralization test.

  9. Activation of PPAR{delta} up-regulates fatty acid oxidation and energy uncoupling genes of mitochondria and reduces palmitate-induced apoptosis in pancreatic {beta}-cells

    Energy Technology Data Exchange (ETDEWEB)

    Wan, Jun; Jiang, Li; Lue, Qingguo; Ke, Linqiu [Department of Endocrinology, West China Hospital of Sichuan University, 37 Guoxue Lane, Chengdu, Sichuan 610041 (China); Li, Xiaoyu [State Key Laboratory of Oral Diseases, Sichuan University, No. 14, 3rd Section, Renmin South Road, Chengdu, Sichuan 610041 (China); Tong, Nanwei, E-mail: buddyjun@hotmail.com [Department of Endocrinology, West China Hospital of Sichuan University, 37 Guoxue Lane, Chengdu, Sichuan 610041 (China)

    2010-01-15

    Recent evidence indicates that decreased oxidative capacity, lipotoxicity, and mitochondrial aberrations contribute to the development of insulin resistance and type 2 diabetes. The goal of this study was to investigate the effects of peroxisome proliferator-activated receptor {delta} (PPAR{delta}) activation on lipid oxidation, mitochondrial function, and insulin secretion in pancreatic {beta}-cells. After HIT-T15 cells (a {beta}-cell line) were exposed to high concentrations of palmitate and GW501516 (GW; a selective agonist of PPAR{delta}), we found that administration of GW increased the expression of PPAR{delta} mRNA. GW-induced activation of PPAR{delta} up-regulated carnitine palmitoyltransferase 1 (CPT1), long-chain acyl-CoA dehydrogenase (LCAD), pyruvate dehydrogenase kinase 4 (PDK4), and uncoupling protein 2 (UCP2); alleviated mitochondrial swelling; attenuated apoptosis; and reduced basal insulin secretion induced by increased palmitate in HIT cells. These results suggest that activation of PPAR{delta} plays an important role in protecting pancreatic {beta}-cells against aberrations caused by lipotoxicity in metabolic syndrome and diabetes.

  10. The avian fossil record in Insular Southeast Asia and its implications for avian biogeography and palaeoecology

    Directory of Open Access Journals (Sweden)

    Hanneke J.M. Meijer

    2014-03-01

    Full Text Available Excavations and studies of existing collections during the last decades have significantly increased the abundance as well as the diversity of the avian fossil record for Insular Southeast Asia. The avian fossil record covers the Eocene through the Holocene, with the majority of bird fossils Pleistocene in age. Fossil bird skeletal remains represent at least 63 species in 54 genera and 27 families, and two ichnospecies are represented by fossil footprints. Birds of prey, owls and swiftlets are common elements. Extinctions seem to have been few, suggesting continuity of avian lineages since at least the Late Pleistocene, although some shifts in species ranges have occurred in response to climatic change. Similarities between the Late Pleistocene avifaunas of Flores and Java suggest a dispersal route across southern Sundaland. Late Pleistocene assemblages of Niah Cave (Borneo and Liang Bua (Flores support the rainforest refugium hypothesis in Southeast Asia as they indicate the persistence of forest cover, at least locally, throughout the Late Pleistocene and Holocene.

  11. The avian tectorial membrane: Why is it tapered?

    CERN Document Server

    Iwasa, Kuni H

    2015-01-01

    While the mammalian- and the avian inner ears have well defined tonotopic organizations as well as hair cells specialized for motile and sensing roles, the structural organization of the avian ear is different from its mammalian cochlear counterpart. Presumably this difference stems from the difference in the way motile hair cells function. Short hair cells, whose role is considered analogous to mammalian outer hair cells, presumably depends on their hair bundles, and not motility of their cell body, in providing the motile elements of the cochlear amplifier. This report focuses on the role of the avian tectorial membrane, specifically by addressing the question, "Why is the avian tectorial membrane tapered from the neural to the abneural direction?"

  12. Historical review of avian botulism at Stillwater Wildlife Management Area

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — The purpose of this report is to review historical information on avian botulism at Stillwater Wildlife Management Area. This report includes incidental reports of...

  13. Region 6 Avian Health Program FY2011 Annual Report

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This report describes activities and fund allocations of the Region 6 Avian Health Program in FY2011. Activities include morbidity and mortality monitoring, disease...

  14. Avian influenza surveillance sample collection and shipment protocol

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Instructions for mortality collection and shipment of avian influenza (AI) live bird surveillance sample collections. AI sample collections will include...

  15. Markov Chain Estimation of Avian Seasonal Fecundity, Presentation

    Science.gov (United States)

    Avian seasonal fecundity is of interest from evolutionary, ecological, and conservation perspectives. However, direct estimation of seasonal fecundity is difficult, especially with multibrooded birds, and models representing the renesting and quitting processes are usually requi...

  16. Avian Point Transect Survey; Seward Peninsula, Alaska, 2012

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — This data product contains avian point-transect survey data and habitat data collected on the Seward Peninsula, Alaska, USA, during 21 May – 10 June 2012. We...

  17. The 3rd International Symposium on Avian Brood Parasitism

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

    正Invited participants on the 3rd International Symposium on Avian Brood Parasitism, sponsored by Hainan Normal University (HNU), China, Norwegian University of Science and Technology (NTNU), Norway, the Research Council of Norway, and China Ornithological Society (COS).

  18. Migratory Bird Avian Influenza Sampling; Yukon Kuskokwim Delta, Alaska, 2015

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Data set containing avian influenza sampling information for spring and summer waterbirds on the Yukon Kuskokwim Delta, 2015. Data contains sample ID, species common...

  19. Avian populations and habitat use in interior Alaska taiga

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Avian community structure, habitat occupancy levels, and species habitat use patterns were examined in the woody habitats of interior Alaska taiga. Some birds...

  20. Avian Flu (H7N9) in China

    Science.gov (United States)

    ... Mobile Apps RSS Feeds Avian Flu (H7N9) in China Recommend on Facebook Tweet Share Compartir Warning - Level ... of H7N9 have been reported outside of mainland China but most of these infections have occurred among ...