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Sample records for grk2 protein levels

  1. GRK2 Fine-Tunes Circadian Clock Speed and Entrainment via Transcriptional and Post-translational Control of PERIOD Proteins

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    Neel Mehta

    2015-08-01

    Full Text Available The pacemaker properties of the suprachiasmatic nucleus (SCN circadian clock are shaped by mechanisms that influence the expression and behavior of clock proteins. Here, we reveal that G-protein-coupled receptor kinase 2 (GRK2 modulates the period, amplitude, and entrainment characteristics of the SCN. Grk2-deficient mice show phase-dependent alterations in light-induced entrainment, slower recovery from jetlag, and longer behavioral rhythms. Grk2 ablation perturbs intrinsic rhythmic properties of the SCN, increasing amplitude and decreasing period. At the cellular level, GRK2 suppresses the transcription of the mPeriod1 gene and the trafficking of PERIOD1 and PERIOD2 proteins to the nucleus. Moreover, GRK2 can physically interact with PERIOD1/2 and promote PERIOD2 phosphorylation at Ser545, effects that may underlie its ability to regulate PERIOD1/2 trafficking. Together, our findings identify GRK2 as an important modulator of circadian clock speed, amplitude, and entrainment by controlling PERIOD at the transcriptional and post-translational levels.

  2. Myocardial Ablation of G Protein-Coupled Receptor Kinase 2 (GRK2 Decreases Ischemia/Reperfusion Injury through an Anti-Intrinsic Apoptotic Pathway.

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    Qian Fan

    Full Text Available Studies from our lab have shown that decreasing myocardial G protein-coupled receptor kinase 2 (GRK2 activity and expression can prevent heart failure progression after myocardial infarction. Since GRK2 appears to also act as a pro-death kinase in myocytes, we investigated the effect of cardiomyocyte-specific GRK2 ablation on the acute response to cardiac ischemia/reperfusion (I/R injury. To do this we utilized two independent lines of GRK2 knockout (KO mice where the GRK2 gene was deleted in only cardiomyocytes either constitutively at birth or in an inducible manner that occurred in adult mice prior to I/R. These GRK2 KO mice and appropriate control mice were subjected to a sham procedure or 30 min of myocardial ischemia via coronary artery ligation followed by 24 hrs reperfusion. Echocardiography and hemodynamic measurements showed significantly improved post-I/R cardiac function in both GRK2 KO lines, which correlated with smaller infarct sizes in GRK2 KO mice compared to controls. Moreover, there was significantly less TUNEL positive myocytes, less caspase-3, and -9 but not caspase-8 activities in GRK2 KO mice compared to control mice after I/R injury. Of note, we found that lowering cardiac GRK2 expression was associated with significantly lower cytosolic cytochrome C levels in both lines of GRK2 KO mice after I/R compared to corresponding control animals. Mechanistically, the anti-apoptotic effects of lowering GRK2 expression were accompanied by increased levels of Bcl-2, Bcl-xl, and increased activation of Akt after I/R injury. These findings were reproduced in vitro in cultured cardiomyocytes and GRK2 mRNA silencing. Therefore, lowering GRK2 expression in cardiomyocytes limits I/R-induced injury and improves post-ischemia recovery by decreasing myocyte apoptosis at least partially via Akt/Bcl-2 mediated mitochondrial protection and implicates mitochondrial-dependent actions, solidifying GRK2 as a pro-death kinase in the heart.

  3. Extracellular signal-regulated kinases control expression of G protein-coupled receptor kinase 2 (GRK2)

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    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...

  4. G protein-coupled receptor kinase-2 (GRK-2) regulates serotonin metabolism through the monoamine oxidase AMX-2 in Caenorhabditis elegans.

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    Wang, Jianjun; Luo, Jiansong; Aryal, Dipendra K; Wetsel, William C; Nass, Richard; Benovic, Jeffrey L

    2017-04-07

    G protein-coupled receptors (GPCRs) regulate many animal behaviors. GPCR signaling is mediated by agonist-promoted interactions of GPCRs with heterotrimeric G proteins, GPCR kinases (GRKs), and arrestins. To further elucidate the role of GRKs in regulating GPCR-mediated behaviors, we utilized the genetic model system Caenorhabditis elegans Our studies demonstrate that grk-2 loss-of-function strains are egg laying-defective and contain low levels of serotonin (5-HT) and high levels of the 5-HT metabolite 5-hydroxyindole acetic acid (5-HIAA). The egg laying defect could be rescued by the expression of wild type but not by catalytically inactive grk-2 or by the selective expression of grk-2 in hermaphrodite-specific neurons. The addition of 5-HT or inhibition of 5-HT metabolism also rescued the egg laying defect. Furthermore, we demonstrate that AMX-2 is the primary monoamine oxidase that metabolizes 5-HT in C. elegans, and we also found that grk-2 loss-of-function strains have abnormally high levels of AMX-2 compared with wild-type nematodes. Interestingly, GRK-2 was also found to interact with and promote the phosphorylation of AMX-2. Additional studies reveal that 5-HIAA functions to inhibit egg laying in a manner dependent on the 5-HT receptor SER-1 and the G protein GOA-1. These results demonstrate that GRK-2 modulates 5-HT metabolism by regulating AMX-2 function and that 5-HIAA may function in the SER-1 signaling pathway. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  5. Sequestration of muscarinic acetylcholine receptor m2 subtypes. Facilitation by G protein-coupled receptor kinase (GRK2) and attenuation by a dominant-negative mutant of GRK2.

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    Tsuga, H; Kameyama, K; Haga, T; Kurose, H; Nagao, T

    1994-12-23

    Sequestration of m2 receptors (muscarinic acetylcholine receptor m2 subtypes), which was assessed as loss of N-[3H]methylscopolamine ([3H]NMS) binding activity from the cell surface, was examined in COS 7 and BHK-21 cells that had been transfected with expression vectors encoding the m2 receptor and, independently, vectors encoding a G protein-coupled receptor kinase (GRK2) (beta-adrenergic receptor kinase 1) or a GRK2 dominant-negative mutant (DN-GRK2). The sequestration of m2 receptors became apparent when the cells were treated with 10(-5) M or higher concentrations of carbamylcholine. In this case, approximately 40% or 20-25% of the [3H]NMS binding sites on COS 7 or BHK-21 cells, respectively, were sequestered with a half-life of 15-25 min. In cells in which GRK2 was also expressed, the sequestration became apparent in the presence of 10(-7) M carbamylcholine. Approximately 40% of the [3H]NMS binding sites on both COS 7 and BHK-21 cells were sequestered in the presence of 10(-6) M or higher concentrations of carbamylcholine. When DN-GRK2 was expressed in COS 7 cells, the proportion of [3H]NMS binding sites sequestered in the presence of 10(-5) M or higher concentrations of carbamylcholine was reduced to 20-30%. These results indicate that the phosphorylation of m2 receptors by GRK2 facilitates their sequestration. These results are in contrast with the absence of a correlation between sequestration and the phosphorylation of beta-adrenergic receptors by the GRK2 and suggests that the consequences of phosphorylation by GRK2 are different for different receptors.

  6. Sequestration of human muscarinic acetylcholine receptor hm1-hm5 subtypes: effect of G protein-coupled receptor kinases GRK2, GRK4, GRK5 and GRK6.

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    Tsuga, H; Okuno, E; Kameyama, K; Haga, T

    1998-03-01

    Sequestration of porcine muscarinic acetylcholine receptor m2 subtypes (m2 receptors) expressed in COS-7 cells is facilitated by coexpression of G protein-coupled receptor kinases 2 (GRK2). We examined the effect of coexpression of GRK2, GRK4 delta, GRK5 and GRK6 on sequestration of human m1-m5 receptors expressed in COS-7 cells, which was assessed as loss of [3H]N-methylscopolamine binding activity from the cell surface. Sequestration of m4 receptors as well as m2 receptors was facilitated by coexpression of GRK2 and attenuated by coexpression of the dominant negative form of GRK2 (DN-GRK2). Sequestration of m3 and m5 receptors also was facilitated by coexpression of GRK2 but not affected by coexpression of DN-GRK2. On the other hand, proportions of sequestered m1 receptors were not significantly different with coexpression of GRK2 and DN-GRK2. GRK4 delta, GRK5 and GRK6 did not facilitate sequestration of m1-m5 receptors in COS-7 cells, except that the sequestration of m2 receptors tended to be facilitated by coexpression of GRK4 delta, GRK5 and GRK6. However, coexpression of GRK4 delta, GRK5, but not GRK6, in BHK-21 cells facilitated sequestration of m2, but not m3, receptors. These results indicate that the effect of GRK2 to facilitate receptor sequestration is not restricted to m2 receptors but is generalized to other muscarinic receptors except m1 receptors and that other kinases, including GRK4 delta, GRK5 and endogenous kinase(s) in COS-7 cells, also contribute to sequestration of m2 and m4 receptors.

  7. GRK2 negatively regulates IGF-1R signaling pathway and cyclins' expression in HepG2 cells.

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    Wei, Zhengyu; Hurtt, Reginald; Gu, Tina; Bodzin, Adam S; Koch, Walter J; Doria, Cataldo

    2013-09-01

    G protein coupled receptor kinase 2 (GRK2) plays a central role in the regulation of a variety of important signaling pathways. Alternation of GRK2 protein level and activity casts profound effects on cell physiological functions and causes diseases such as heart failure, rheumatoid arthritis, and obesity. We have previously reported that overexpression of GRK2 has an inhibitory role in cancer cell growth. To further examine the role of GRK2 in cancer, in this study, we investigated the effects of reduced protein level of GRK2 on insulin-like growth factor 1 receptor (IGF-1R) signaling pathway in human hepatocellular carcinoma (HCC) HepG2 cells. We created a GRK2 knockdown cell line using a lentiviral vector mediated expression of GRK2 specific short hairpin RNA (shRNA). Under IGF-1 stimulation, HepG2 cells with reduced level of GRK2 showed elevated total IGF-1R protein expression as well as tyrosine phosphorylation of receptor. In addition, HepG2 cells with reduced level of GRK2 also demonstrated increased tyrosine phosphorylation of IRS1 at the residue 612 and increased phosphorylation of Akt, indicating a stronger activation of IGF-1R signaling pathway. However, HepG2 cells with reduced level of GRK2 did not display any growth advantage in culture as compared with the scramble control cells. We further detected that reduced level of GRK2 induced a small cell cycle arrest at G2/M phase by enhancing the expression of cyclin A, B1, and E. Our results indicate that GRK2 has contrasting roles on HepG2 cell growth by negatively regulating the IGF-1R signaling pathway and cyclins' expression.

  8. GRK2 protein-mediated transphosphorylation contributes to loss of function of μ-opioid receptors induced by neuropeptide FF (NPFF2) receptors.

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    Moulédous, Lionel; Froment, Carine; Dauvillier, Stéphanie; Burlet-Schiltz, Odile; Zajac, Jean-Marie; Mollereau, Catherine

    2012-04-13

    Neuropeptide FF (NPFF) interacts with specific receptors to modulate opioid functions in the central nervous system. On dissociated neurons and neuroblastoma cells (SH-SY5Y) transfected with NPFF receptors, NPFF acts as a functional antagonist of μ-opioid (MOP) receptors by attenuating the opioid-induced inhibition of calcium conductance. In the SH-SY5Y model, MOP and NPFF(2) receptors have been shown to heteromerize. To understand the molecular mechanism involved in the anti-opioid activity of NPFF, we have investigated the phosphorylation status of the MOP receptor using phospho-specific antibody and mass spectrometry. Similarly to direct opioid receptor stimulation, activation of the NPFF(2) receptor by [D-Tyr-1-(NMe)Phe-3]NPFF (1DMe), an analog of NPFF, induced the phosphorylation of Ser-377 of the human MOP receptor. This heterologous phosphorylation was unaffected by inhibition of second messenger-dependent kinases and, contrarily to homologous phosphorylation, was prevented by inactivation of G(i/o) proteins by pertussis toxin. Using siRNA knockdown we could demonstrate that 1DMe-induced Ser-377 cross-phosphorylation and MOP receptor loss of function were mediated by the G protein receptor kinase GRK2. In addition, mass spectrometric analysis revealed that the phosphorylation pattern of MOP receptors was qualitatively similar after treatment with the MOP agonist Tyr-D-Ala-Gly (NMe)-Phe-Gly-ol (DAMGO) or after treatment with the NPFF agonist 1DMe, but the level of multiple phosphorylation was more intense after DAMGO. Finally, NPFF(2) receptor activation was sufficient to recruit β-arrestin2 to the MOP receptor but not to induce its internalization. These data show that NPFF-induced heterologous desensitization of MOP receptor signaling is mediated by GRK2 and could involve transphosphorylation within the heteromeric receptor complex.

  9. Reduction of sympathetic activity via adrenal-targeted GRK2 gene deletion attenuates heart failure progression and improves cardiac function after myocardial infarction.

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    Lymperopoulos, Anastasios; Rengo, Giuseppe; Gao, Erhe; Ebert, Steven N; Dorn, Gerald W; Koch, Walter J

    2010-05-21

    Chronic heart failure (HF) is characterized by sympathetic overactivity and enhanced circulating catecholamines (CAs), which significantly increase HF morbidity and mortality. We recently reported that adrenal G protein-coupled receptor kinase 2 (GRK2) is up-regulated in chronic HF, leading to enhanced CA release via desensitization/down-regulation of the chromaffin cell alpha(2)-adrenergic receptors that normally inhibit CA secretion. We also showed that adrenal GRK2 inhibition decreases circulating CAs and improves cardiac inotropic reserve and function. Herein, we hypothesized that adrenal-targeted GRK2 gene deletion before the onset of HF might be beneficial by reducing sympathetic activation. To specifically delete GRK2 in the chromaffin cells of the adrenal gland, we crossed PNMTCre mice, expressing Cre recombinase under the chromaffin cell-specific phenylethanolamine N-methyltransferase (PNMT) gene promoter, with floxedGRK2 mice. After confirming a significant ( approximately 50%) reduction of adrenal GRK2 mRNA and protein levels, the PNMT-driven GRK2 knock-out (KO) offspring underwent myocardial infarction (MI) to induce HF. At 4 weeks post-MI, plasma levels of both norepinephrine and epinephrine were reduced in PNMT-driven GRK2 KO, compared with control mice, suggesting markedly reduced post-MI sympathetic activation. This translated in PNMT-driven GRK2 KO mice into improved cardiac function and dimensions as well as amelioration of abnormal cardiac beta-adrenergic receptor signaling at 4 weeks post-MI. Thus, adrenal-targeted GRK2 gene KO decreases circulating CAs, leading to improved cardiac function and beta-adrenergic reserve in post-MI HF. GRK2 inhibition in the adrenal gland might represent a novel sympatholytic strategy that can aid in blocking HF progression.

  10. GRK2 deficiency is a pain

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    Kleibeuker, W.

    2007-01-01

    G protein-coupled receptor (GPCR) kinase (GRK)2 regulates the responsiveness of many GPCRS, e.g. CCR1, CCR2, CCR5, neurokinin-1 receptor, and metabotropic glutamate receptors, which are involved in the development of allodynia (pain due to a stimulus which does normally not provoke pain) and hyperal

  11. Adrenal GRK2 upregulation mediates sympathetic overdrive in heart failure.

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    Lymperopoulos, Anastasios; Rengo, Giuseppe; Funakoshi, Hajime; Eckhart, Andrea D; Koch, Walter J

    2007-03-01

    Cardiac overstimulation by the sympathetic nervous system (SNS) is a salient characteristic of heart failure, reflected by elevated circulating levels of catecholamines. The success of beta-adrenergic receptor (betaAR) antagonists in heart failure argues for SNS hyperactivity being pathogenic; however, sympatholytic agents targeting alpha2AR-mediated catecholamine inhibition have been unsuccessful. By investigating adrenal adrenergic receptor signaling in heart failure models, we found molecular mechanisms to explain the failure of sympatholytic agents and discovered a new strategy to lower SNS activity. During heart failure, there is substantial alpha2AR dysregulation in the adrenal gland, triggered by increased expression and activity of G protein-coupled receptor kinase 2 (GRK2). Adrenal gland-specific GRK2 inhibition reversed alpha2AR dysregulation in heart failure, resulting in lowered plasma catecholamine levels, improved cardiac betaAR signaling and function, and increased sympatholytic efficacy of a alpha2AR agonist. This is the first demonstration, to our knowledge, of a molecular mechanism for SNS hyperactivity in heart failure, and our study identifies adrenal GRK2 activity as a new sympatholytic target.

  12. GRK2: putting the brakes on the circadian clock

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    Mendoza-Viveros, Lucia; Cheng, Arthur H.

    2016-01-01

    G protein-coupled receptor kinases (GRKs) are a family of serine/threonine protein kinases that terminate G protein-coupled receptor (GPCR) signaling by phosphorylating the receptor and inducing its internalization. In addition to their canonical function, some GRKs can phosphorylate non-GPCR substrates and regulate GPCR signaling in a kinase-independent manner. GPCRs are abundantly expressed in the suprachiasmatic nucleus (SCN), a structure in the mammalian brain that serves as the central circadian pacemaker. Various facets of circadian timekeeping are under the influence of GPCR signaling, and thus are potential targets for GRK regulation. Despite this, little attention has been given to the role of GRKs in circadian rhythms. In this research highlight, we discuss our latest findings on the functional involvement of GRK2 in mammalian circadian timekeeping in the SCN. Using grk2 knockout mice, we demonstrate that GRK2 is critical for maintaining proper clock speed and ensuring that the clock is appropriately synchronized to environmental light cycles. Although grk2 deficiency expectedly alters the expression of a key GPCR in the SCN, our study also reveals that GRK2 has a more direct function that touches the heart of the circadian clock. PMID:27088110

  13. Regulation of FcϵRI Signaling in Mast Cells by G Protein-coupled Receptor Kinase 2 and Its RH Domain*

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    Subramanian, Hariharan; Gupta, Kshitij; Parameswaran, Narayanan; Ali, Hydar

    2014-01-01

    Agonist-induced phosphorylation of G protein-coupled receptors (GPCRs) by GPCR kinases (GRKs) promotes their desensitization and internalization. Here, we sought to determine the role of GRK2 on FcϵRI signaling and mediator release in mast cells. The strategies utilized included lentiviral shRNA-mediated GRK2 knockdown, GRK2 gene deletion (GRK2flox/flox/cre recombinase) and overexpression of GRK2 and its regulator of G protein signaling homology (RH) domain (GRK2-RH). We found that silencing GRK2 expression caused ∼50% decrease in antigen-induced Ca2+ mobilization and degranulation but resulted in ablation of cytokine (IL-6 and IL-13) generation. The effect of GRK2 on cytokine generation does not require its catalytic activity but is mediated via the phosphorylation of p38 and Akt. Overexpression of GRK2 or its RH domain (GRK2-RH) enhanced antigen-induced mast cell degranulation and cytokine generation without affecting the expression levels of any of the FcϵRI subunits (α, β, and γ). GRK2 or GRK2-RH had no effect on antigen-induced phosphorylation of FcϵRIγ or Src but enhanced tyrosine phosphorylation of Syk. These data demonstrate that GRK2 modulates FcϵRI signaling in mast cells via at least two mechanisms. One involves GRK2-RH and modulates tyrosine phosphorylation of Syk, and the other is mediated via the phosphorylation of p38 and Akt. PMID:24904059

  14. Effects of paroxetine-mediated inhibition of GRK2 expression on depression and cardiovascular function in patients with myocardial infarction

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    Tian XQ

    2016-09-01

    Full Text Available Xiuqing Tian,1 Qing Wang,1 Rui Guo,2 Lingling Xu,3 Qin M Chen,4 Yinglong Hou1 1Department of Cardiology, Qianfoshan Hospital of Shandong University, Jinan City, 2Department of Physiology, Nanjing University of Chinese Medicine, Nanjing City, 3College of Medicine, Weifang Medical University, Weifang City, People’s Republic of China; 4Department of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ, USA Background: Paroxetine is a selective serotonin reuptake inhibitor utilized in the treatment of depression and anxiety disorders. Recent studies have identified paroxetine as a G protein-coupled receptor kinase-2 (GRK2 inhibitor capable of reversing cardiac dysfunction and remodeling in experimental models of acute myocardial infarction (AMI. We determine the clinical importance of paroxetine on cardiac functions in patients having AMI with depression (AMID in comparison with fluoxetine, an unrelated selective serotonin reuptake inhibitor that does not inhibit GRK2.Methods: Diagnosis of depression was based on the 17-item Hamilton Depression Scale and Self-rating Depression Scale in AMI patients after hospital admission. AMID patients were randomly assigned to paroxetine or fluoxetine for treatment of depression. Heart rate variability and cardiac function were evaluated. GRK2 protein levels were measured using peripheral lymphocytes and Western blot.Results: GRK2 expression in AMID patients was significantly higher than that in AMI patients without depression. In AMID patients, GRK2 levels were positively correlated with the 17-item Hamilton Depression Scale and the Self-rating Depression Scale scores, and negatively correlated with heart rate variability. Treatment of AMID patients with paroxetine significantly reduced the expression of GRK2, normalized the autonomic nervous system function, and improved cardiac performance. In contrast, fluoxetine normalized the autonomic nervous system but did not reduce the expression of

  15. Differential interaction of GRK2 with members of the G alpha q family.

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    Day, Peter W; Carman, Christopher V; Sterne-Marr, Rachel; Benovic, Jeffrey L; Wedegaertner, Philip B

    2003-08-05

    Regulators of G protein signaling (RGS) proteins bind to active G alpha subunits and accelerate the rate of GTP hydrolysis and/or block interaction with effector molecules, thereby decreasing signal duration and strength. RGS proteins are defined by the presence of a conserved 120-residue region termed the RGS domain. Recently, it was shown that the G protein-coupled receptor kinase 2 (GRK2) contains an RGS domain that binds to the active form of G alpha(q). Here, the ability of GRK2 to interact with other members of the G alpha(q) family, G alpha(11), G alpha(14), and G alpha(16), was tested. The signaling of all members of the G alpha(q) family, with the exception of G alpha(16), was inhibited by GRK2. Immunoprecipitation of full-length GRK2 or pull down of GST-GRK2-(45-178) resulted in the detection of G alpha(q), but not G alpha(16), in an activation-dependent manner. Moreover, activated G alpha(16) failed to promote plasma membrane (PM) recruitment of a GRK2-(45-178)-GFP fusion protein. Assays with chimeric G alpha(q)(-)(16) subunits indicated that the C-terminus of G alpha(q) mediates binding to GRK2. Despite showing no interaction with GRK2, G alpha(16) does interact with RGS2, in both inositol phosphate and PM recruitment assays. Thus, GRK2 is the first identified RGS protein that discriminates between members of the G alpha(q) family, while another RGS protein, RGS2, binds to both G alpha(q) and G alpha(16).

  16. Regulation of Constitutive GPR3 Signaling and Surface Localization by GRK2 and β-arrestin-2 Overexpression in HEK293 Cells.

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    Katie M Lowther

    Full Text Available G protein-coupled receptor 3 (GPR3 is a constitutively active receptor that maintains high 3'-5'-cyclic adenosine monophosphate (cAMP levels required for meiotic arrest in oocytes and CNS function. Ligand-activated G protein-coupled receptors (GPCRs signal at the cell surface and are silenced by phosphorylation and β-arrestin recruitment upon endocytosis. Some GPCRs can also signal from endosomes following internalization. Little is known about the localization, signaling, and regulation of constitutively active GPCRs. We demonstrate herein that exogenously-expressed GPR3 localizes to the cell membrane and undergoes internalization in HEK293 cells. Inhibition of endocytosis increased cell surface-localized GPR3 and cAMP levels while overexpression of GPCR-Kinase 2 (GRK2 and β-arrestin-2 decreased cell surface-localized GPR3 and cAMP levels. GRK2 by itself is sufficient to decrease cAMP production but both GRK2 and β-arrestin-2 are required to decrease cell surface GPR3. GRK2 regulates GPR3 independently of its kinase activity since a kinase inactive GRK2-K220R mutant significantly decreased cAMP levels. However, GRK2-K220R and β-arrestin-2 do not diminish cell surface GPR3, suggesting that phosphorylation is required to induce GPR3 internalization. To understand which residues are targeted for desensitization, we mutated potential phosphorylation sites in the third intracellular loop and C-terminus and examined the effect on cAMP and receptor surface localization. Mutation of residues in the third intracellular loop dramatically increased cAMP levels whereas mutation of residues in the C-terminus produced cAMP levels comparable to GPR3 wild type. Interestingly, both mutations significantly reduced cell surface expression of GPR3. These results demonstrate that GPR3 signals at the plasma membrane and can be silenced by GRK2/β-arrestin overexpression. These results also strongly implicate the serine and/or threonine residues in the third

  17. Skeletal Muscle-specific G Protein-coupled Receptor Kinase 2 Ablation Alters Isolated Skeletal Muscle Mechanics and Enhances Clenbuterol-stimulated Hypertrophy.

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    Woodall, Benjamin P; Woodall, Meryl C; Luongo, Timothy S; Grisanti, Laurel A; Tilley, Douglas G; Elrod, John W; Koch, Walter J

    2016-10-14

    GRK2, a G protein-coupled receptor kinase, plays a critical role in cardiac physiology. Adrenergic receptors are the primary target for GRK2 activity in the heart; phosphorylation by GRK2 leads to desensitization of these receptors. As such, levels of GRK2 activity in the heart directly correlate with cardiac contractile function. Furthermore, increased expression of GRK2 after cardiac insult exacerbates injury and speeds progression to heart failure. Despite the importance of this kinase in both the physiology and pathophysiology of the heart, relatively little is known about the role of GRK2 in skeletal muscle function and disease. In this study we generated a novel skeletal muscle-specific GRK2 knock-out (KO) mouse (MLC-Cre:GRK2(fl/fl)) to gain a better understanding of the role of GRK2 in skeletal muscle physiology. In isolated muscle mechanics testing, GRK2 ablation caused a significant decrease in the specific force of contraction of the fast-twitch extensor digitorum longus muscle yet had no effect on the slow-twitch soleus muscle. Despite these effects in isolated muscle, exercise capacity was not altered in MLC-Cre:GRK2(fl/fl) mice compared with wild-type controls. Skeletal muscle hypertrophy stimulated by clenbuterol, a β2-adrenergic receptor (β2AR) agonist, was significantly enhanced in MLC-Cre:GRK2(fl/fl) mice; mechanistically, this seems to be due to increased clenbuterol-stimulated pro-hypertrophic Akt signaling in the GRK2 KO skeletal muscle. In summary, our study provides the first insights into the role of GRK2 in skeletal muscle physiology and points to a role for GRK2 as a modulator of contractile properties in skeletal muscle as well as β2AR-induced hypertrophy. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  18. GRK2 and chronic pain

    NARCIS (Netherlands)

    Wang, H.

    2012-01-01

    G protein coupled receptor kinases (GRKs) regulate the termination of G protein-coupled receptor (GPCR) activation by phosphorylating agonist-occupied GPCRs. Apart from terminating GPCR activation, more evidence emerges in recent years demonstrating that GRKs also interact with multiple intracellula

  19. Reversal of diet-induced obesity and insulin resistance by inducible genetic ablation of GRK2

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    Vila-Bedmar, Rocio; Cruces-Sande, Marta; Lucas, Elisa; Willemen, Hanneke L D M; Heijnen, Cobi J; Kavelaars, Annemieke; Mayor, Federico; Murga, Cristina

    2015-01-01

    Insulin resistance is a common feature of obesity and predisposes individuals to various prevalent pathological conditions. G protein (heterotrimeric guanine nucleotide-binding protein)-coupled receptor kinase 2 (GRK2) integrates several signal transduction pathways and is emerging as a physiologica

  20. Sonic Hedgehog dependent phosphorylation by CK1α and GRK2 is required for ciliary accumulation and activation of smoothened.

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    Yongbin Chen

    2011-06-01

    Full Text Available Hedgehog (Hh signaling regulates embryonic development and adult tissue homeostasis through the GPCR-like protein Smoothened (Smo, but how vertebrate Smo is activated remains poorly understood. In Drosophila, Hh dependent phosphorylation activates Smo. Whether this is also the case in vertebrates is unclear, owing to the marked sequence divergence between vertebrate and Drosophila Smo (dSmo and the involvement of primary cilia in vertebrate Hh signaling. Here we demonstrate that mammalian Smo (mSmo is activated through multi-site phosphorylation of its carboxyl-terminal tail by CK1α and GRK2. Phosphorylation of mSmo induces its active conformation and simultaneously promotes its ciliary accumulation. We demonstrate that graded Hh signals induce increasing levels of mSmo phosphorylation that fine-tune its ciliary localization, conformation, and activity. We show that mSmo phosphorylation is induced by its agonists and oncogenic mutations but is blocked by its antagonist cyclopamine, and efficient mSmo phosphorylation depends on the kinesin-II ciliary motor. Furthermore, we provide evidence that Hh signaling recruits CK1α to initiate mSmo phosphorylation, and phosphorylation further increases the binding of CK1α and GRK2 to mSmo, forming a positive feedback loop that amplifies and/or sustains mSmo phosphorylation. Hence, despite divergence in their primary sequences and their subcellular trafficking, mSmo and dSmo employ analogous mechanisms for their activation.

  1. Beta-blockers decreases Th17/Treg cytokine ratios by down regulation of GRK2 synthesis in lymphocytes of patients with ACS%β受体阻滞剂对急性冠脉综合征Th17/Treg的作用研究

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    田秀青; 王清; 葛庆玲

    2016-01-01

    Objective To observe the effects of g protein-coupled receptor kinase-2 (GRK2) in peripheral lymphocytes on Th17/Treg cytokine ratios and investigate the potential impact and mechanism of beta-blockers on immune function in patients with acute coronary syndrome (ACS). Methods One hundred-four patients with ACS and forty-five normal coronary angiography (NCA) hospitalized patients as control group were enrolled in the study. ACS group was randomly divided into metoprolol group(n=52) and carvedilol group(n=52).The levels of lymphocyte GRK2 were detected by means of a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) and IL-17 and FoxP3 in lymphocytes using flow cytometry at baseline and 6 months after the treatment. Results The levels of lymphocyte GRK2 and Th17/Treg in ACS patients were significantly higher than those of the UCA controls (both P0.05). Conclusion These results suggest that the increase of GRK2 may affect GRK2 in peripheral lymphocytes of ACS. Beta-blockers appear to have a beneficial effect on the Th17/Treg balance of immune system through decreasing activities of GRK2.%目的:探讨β受体阻滞剂对急性冠脉综合征(ACS)患者淋巴细胞G蛋白偶联受体激酶2(GRK2)和Th17/Treg的作用及意义。方法选择2013年2月至2015年12月千佛山医院心内科收治的CHD患者104例以及非CHD住院患者35例(NCA组)作为研究对象。ACS患者随机分为美托洛尔和卡维地洛治疗组。流式细胞仪测定淋巴细胞IL-17和Foxp3水平,酶联免疫吸附法测定GRK2水平。随访6个月。结果 ACS患者GRK2及IL-17/Foxp3比值均较非冠心病组明显升高(P0.05)。GRK2与IL-17/FoxP3比值呈显著正相关(P<0.01)。结论 ACS患者Th17/Treg失调与GRK2升高有关。β受体阻滞剂可降低GRK2水平,改善Th17/Treg平衡,具有免疫调节作用。

  2. Critical role for Epac1 in inflammatory pain controlled by GRK2-mediated phosphorylation of Epac1

    NARCIS (Netherlands)

    Singhmar, Pooja; Huo, XiaoJiao; Eijkelkamp, Niels; Berciano, Susana Rojo; Baameur, Faiza; Mei, Fang C; Zhu, Yingmin; Cheng, Xiaodong; Hawke, David; Mayor, Federico; Murga, Cristina; Heijnen, Cobi J.; Kavelaars, Annemieke

    2016-01-01

    cAMP signaling plays a key role in regulating pain sensitivity. Here, we uncover a previously unidentified molecular mechanism in which direct phosphorylation of the exchange protein directly activated by cAMP 1 (EPAC1) by G protein kinase 2 (GRK2) suppresses Epac1-to-Rap1 signaling, thereby

  3. Modulation of adrenal catecholamine secretion by in vivo gene transfer and manipulation of G protein-coupled receptor kinase-2 activity.

    Science.gov (United States)

    Lymperopoulos, Anastasios; Rengo, Giuseppe; Zincarelli, Carmela; Soltys, Stephen; Koch, Walter J

    2008-02-01

    We recently reported that the upregulation of adrenal G protein-coupled receptor kinase-2 (GRK2) causes enhanced catecholamine (CA) secretion by desensitizing sympatho-inhibitory alpha (2)-adrenergic receptors (alpha (2)ARs) of chromaffin cells, and thereby aggravating heart failure (HF). In this study, we sought to develop an efficient and reproducible in vivo adrenal gene transfer method to determine whether manipulation of adrenal GRK2 levels/activity regulates physiological CA secretion in rats. We specifically investigated two different in vivo gene delivery methods: direct injection into the suprarenal glands, and retrograde delivery through the suprarenal veins. We delivered adenoviral (Ad) vectors containing either GRK2 or an inhibitor of GRK2 activity, the beta ARKct. We found both delivery approaches equally effective at supporting robust (>80% of the whole organ) and adrenal-restricted transgene expression, in the cortical region as well as in the medullar region. Additionally, rats with AdGRK2-infected adrenals exhibit enhanced plasma CA levels when compared with control rats (AdGFP-injected adrenals), whereas plasma CA levels after Ad beta ARKct infection were significantly lower. Finally, in isolated chromaffin cells, alpha (2)ARs of AdGRK2-infected cells failed to inhibit CA secretion whereas Ad beta ARKct-infected cells showed normal alpha (2)AR responsiveness. These results not only indicate that in vivo adrenal gene transfer is an effective way of manipulating adrenal gland signalling, but also identify GRK2 as a critically important molecule involved in CA secretion.

  4. Molecular physiopathology of obesity-related diseases: multi-organ integration by GRK2.

    Science.gov (United States)

    Lucas, Elisa; Cruces-Sande, Marta; Briones, Ana M; Salaices, Mercedes; Mayor, Federico; Murga, Cristina; Vila-Bedmar, Rocio

    2015-01-01

    Obesity is a worldwide problem that has reached epidemic proportions both in developed and developing countries. The excessive accumulation of fat poses a risk to health since it favours the development of metabolic alterations including insulin resistance and tissue inflammation, which further contribute to the progress of the complex pathological scenario observed in the obese. In this review we put together the different outcomes of fat accumulation and insulin resistance in the main insulin-responsive tissues, and discuss the role of some of the key molecular routes that control disease progression both in an organ-specific and also in a more systemic manner. In particular, we focus on the importance of studying the integrated regulation of different organs and pathways that contribute to the global pathophysiology of this condition with a specific emphasis on the role of emerging key molecular nodes such as the G protein-coupled receptor kinase 2 (GRK2) signalling hub.

  5. The GIP receptor displays higher basal activity than the GLP-1 receptor but does not recruit GRK2 or arrestin3 effectively.

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    Suleiman Al-Sabah

    Full Text Available Glucagon-like peptide-1 (GLP-1 and glucose-dependent insulinotropic polypeptide (GIP are important regulators of insulin secretion, and their functional loss is an early characteristic of type 2 diabetes mellitus (T2DM. Pharmacological levels of GLP-1, but not GIP, can overcome this loss. GLP-1 and GIP exert their insulinotropic effects through their respective receptors expressed on pancreatic β-cells. Both the GLP-1 receptor (GLP-1R and the GIP receptor (GIPR are members of the secretin family of G protein-coupled receptors (GPCRs and couple positively to adenylate cyclase. We compared the signalling properties of these two receptors to gain further insight into why GLP-1, but not GIP, remains insulinotropic in T2DM patients.GLP-1R and GIPR were transiently expressed in HEK-293 cells, and basal and ligand-induced cAMP production were investigated using a cAMP-responsive luciferase reporter gene assay. Arrestin3 (Arr3 recruitment to the two receptors was investigated using enzyme fragment complementation, confocal microscopy and fluorescence resonance energy transfer (FRET.GIPR displayed significantly higher (P<0.05 ligand-independent activity than GLP-1R. Arr3 displayed a robust translocation to agonist-stimulated GLP-1R but not to GIPR. These observations were confirmed in FRET experiments, in which GLP-1 stimulated the recruitment of both GPCR kinase 2 (GRK2 and Arr3 to GLP-1R. These interactions were not reversed upon agonist washout. In contrast, GIP did not stimulate recruitment of either GRK2 or Arr3 to its receptor. Interestingly, arrestin remained at the plasma membrane even after prolonged (30 min stimulation with GLP-1. Although the GLP-1R/arrestin interaction could not be reversed by agonist washout, GLP-1R and arrestin did not co-internalise, suggesting that GLP-1R is a class A receptor with regard to arrestin binding.GIPR displays higher basal activity than GLP-1R but does not effectively recruit GRK2 or Arr3.

  6. Vasopressin V1A receptor mediates cell proliferation through GRK2-EGFR-ERK1/2 pathway in A7r5 cells.

    Science.gov (United States)

    Zhang, Lingling; Wang, Xiaojun; Cao, Hong; Chen, Yunxuan; Chen, Xianfan; Zhao, Xi; Xu, Feifei; Wang, Yifan; Woo, Anthony Yiu-Ho; Zhu, Weizhong

    2016-12-05

    Abnormal proliferation and hypertrophy of vascular smooth muscle (VSMC), as the main structural component of the vasculature, is an important pathological mechanism of hypertension. Recently, increased levels of arginine vasopressin (AVP) and copeptin, the C-terminal fragment of provasopressin, have been shown to correlate with the development of preeclampsia. AVP targets on the Gq-coupled vasopressin V1A receptor and the Gs-coupled V2 receptor in VSMC and the kidneys to regulate vascular tone and water homeostasis. However, the role of the vasopressin receptor on VSM cell proliferation during vascular remodeling is unclear. Here, we studied the effects of AVP on the proliferation of the rat VSMC-derived A7r5 cells. AVP, in a time- and concentration-dependent manner, promoted A7r5 cell proliferation as indicated by the induction of proliferating cell nuclear antigen expression, methylthiazolyldiphenyl-tetrazolium reduction and incorporation of 5'-bromodeoxyuridine into cellular DNA. These effects, coupled with the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), were blocked by a V1A receptor antagonist SR45059 but not by a V2 receptor antagonist lixivaptan. Although acute activation of V1A receptor induced ERK1/2 phosphorylation via a protein kinase C-dependent pathway, this effect was not involved in cell proliferation. Cell proliferation and ERK1/2 phosphorylation in response to prolonged stimulation with AVP were abolished by inhibition of G protein-coupled receptor kinase 2 (GRK2) and epidermal growth factor receptor (EGFR) using specific inhibitors or small hairpin RNA knock-down. These results suggest that activation of V1A, but not V2 receptor, produces a cell proliferative signal in A7r5 cells via a GRK2/EGFR/ERK1/2-dependent mechanism.

  7. β-Arrestin 1 and 2 and G protein-coupled receptor kinase 2 expression in pituitary adenomas: role in the regulation of response to somatostatin analogue treatment in patients with acromegaly.

    Science.gov (United States)

    Gatto, Federico; Feelders, Richard; van der Pas, Rob; Kros, Johan M; Dogan, Fadime; van Koetsveld, Peter M; van der Lelij, Aart-Jan; Neggers, Sebastian J C M M; Minuto, Francesco; de Herder, Wouter; Lamberts, Steven W J; Ferone, Diego; Hofland, Leo J

    2013-12-01

    Recent in vitro studies highlighted G protein-coupled receptor kinase (GRK)2 and β-arrestins as important players in driving somatostatin receptor (SSTR) desensitization and trafficking. Our aim was to characterize GRK2 and β-arrestins expression in different pituitary adenomas and to investigate their potential role in the response to somatostatin analog (SSA) treatment in GH-secreting adenomas (GHomas). We evaluated mRNA expression of multiple SSTRs, GRK2, β-arrestin 1, and β-arrestin 2 in 41 pituitary adenomas (31 GHomas, 6 nonfunctioning [NFPAs], and 4 prolactinomas [PRLomas]). Within the GHomas group, mRNA data were correlated with the in vivo response to an acute octreotide test and with the GH-lowering effect of SSA in cultured primary cells. β-Arrestin 1 expression was low in all 3 adenoma histotypes. However, its expression was significantly lower in GHomas and PRLomas, compared with NFPAs (P affect GRK2 and β-arrestin expression in GHomas or in cultured rat pituitary tumor GH3 cells. Noteworthy, β-arrestin 1 was significantly lower (P < .05) in tumors responsive to octreotide treatment in vitro, whereas GRK2 and SSTR subtype 2 were significantly higher (P < .05). Likewise, β-arrestin 1 levels were inversely correlated with the in vivo response to acute octreotide test (P = .001), whereas GRK2 and SSTR subtype 2 expression were positively correlated (P < .05). In conclusion, for the first time, we characterized GRK2, β-arrestin 1, and β-arrestin 2 expression in a representative number of pituitary adenomas. β-Arrestin 1 and GRK2 seem to have a role in modulating GH secretion during SSA treatment.

  8. G-protein-coupled receptor kinase 2 and endothelial dysfunction: molecular insights and pathophysiological mechanisms.

    Science.gov (United States)

    Taguchi, Kumiko; Matsumoto, Takayuki; Kobayashi, Tsuneo

    2015-01-01

    Smooth muscle cells (SMC) and endothelial cells are the major cell types in blood vessels. The principal function of vascular SMC in the body is to regulate blood flow and pressure through contraction and relaxation. The endothelium performs a crucial role in maintaining vascular integrity by achieving whole-organ metabolic homeostasis via the production of factors associated with vasoconstriction or vasorelaxation. In this review, we have focused on the production of nitric oxide (NO), a vasorelaxation factor. The extent of NO production represents a key marker in vascular health. A decrease in NO is capable of inducing pathological conditions associated with endothelial dysfunction, such as obesity, diabetes, cardiovascular disease, and atherosclerosis. Recent studies have strongly implicated the involvement of G-protein-coupled receptor kinase 2 (GRK2) in the progression of cardiovascular disease. Vasculature which is affected by insulin resistance and type 2 diabetes expresses high levels of GRK2, which may induce endothelial dysfunction by reducing intracellular NO. GRK2 activation also induces changes in the subcellular localization of GRK2 itself and also of β-arrestin 2, a downstream protein. In this review, we describe the pathophysiological mechanisms of insulin resistance and diabetes, focusing on the signal transduction for NO production via GRK2 and β-arrestin 2, providing novel insights into the potential field of translational investigation in the treatment of diabetic complications.

  9. 骨癌痛大鼠DRG神经元GRK2和β-arrestin2表达以及NGF调节作用的研究%Expression of GRK2 and β-arrestin2 in the dorsal root ganglion neurons and the regulated effect by nerve growth factor in rats with bone cancer pain

    Institute of Scientific and Technical Information of China (English)

    姚鹏; 王志彬; 蒋晶晶; 张锦; 孟凌新

    2011-01-01

    目的:观察大鼠骨癌痛时脊髓背根神经节(DRG)G蛋白偶联受体激酶2(GRK2)和β-arrestin2的变化,探讨鞘内注射抗神经生长因子抗体(anti-NGF)对其表达及疼痛行为学的影响.方法:60只雌性SD大鼠随机分为假手术组、骨癌痛组及骨癌痛+anti-NGF组,13 d后鞘内置管,16 d开始鞘内注入生理盐水或anti-NGF不同时点观察疼痛行为学变化;21 d取同侧L4、L5 DRG,检测β-arrestin2、GRK2蛋白及mRNA表达变化.结果:与假手术组比较,骨癌痛组大鼠体质量减轻[(219±4.8)vs(243±8.1)],自发缩足次数增多[(24.1±3.6)vs(2.9±0.4)],热辐射潜伏期(PWL)缩短[(3.8±0.5)vs(10.9±1.3)],机械痛阈(PWT)降低[(3.2±1.1)vs(12.3±1.3)];与骨癌痛组比较,骨癌痛+anti-NGF组大鼠缩足次数减少(6.7±1.2),PWL延长(9.7±1.2),PWT增高(9.7±1.5).骨癌痛组大鼠β-arrestin2、GRK2表达均高于假手术组,而骨癌痛+anti-NGF组则明显低于骨癌痛组.骨癌痛组大鼠DRG神经元β-arrestin2与GRK2 mRNA的表达均高于假手术组,而骨癌痛+anti-NGF组则均低于骨癌痛组.结论:大鼠骨癌痛时DRG神经元GRK2和β-arrestin2的表达增加,anti-NGF可明显缓解骨癌痛,并对GRK2和β-arrestin2具有调制作用.%OBJECTIVE: To observe the expression of β-arrestin2 and G protein-coupled receptor kinases 2 (GRK2) in the dorsal root ganglion(DRG) neurons, and further investigate the regulated effects by intrathecal application of anti-NGF on the expression and pain-related behavior in rats with bone cancer pain.METHODS: Sixty female rats were divided into sham, cancer and cancer+ anti-NGF group.Bone cancer pain rats were induced by implantation of Walker 256 breast carcinosarcoma cells into the tibia.Each rat was surgically fitted with an intrathecal catheter at days 13, Sodium chloride (groups sham and cancer) or anti-NGF(group eancer+anti-NGF) 10 μL was injected by intrathecal catheter from 16 to 21 days, pain-related behavior were assessed.Western blotting

  10. Cardiac hyporesponsiveness in severe sepsis is associated with nitric oxide-dependent activation of G protein receptor kinase.

    Science.gov (United States)

    Dal-Secco, Daniela; DalBó, Silvia; Lautherbach, Natalia E S; Gava, Fábio N; Celes, Mara R N; Benedet, Patricia O; Souza, Adriana H; Akinaga, Juliana; Lima, Vanessa; Silva, Katiussia P; Kiguti, Luiz Ricardo A; Rossi, Marcos A; Kettelhut, Isis C; Pupo, André S; Cunha, Fernando Q; Assreuy, Jamil

    2017-07-01

    G protein-coupled receptor kinase isoform 2 (GRK2) has a critical role in physiological and pharmacological responses to endogenous and exogenous substances. Sepsis causes an important cardiovascular dysfunction in which nitric oxide (NO) has a relevant role. The present study aimed to assess the putative effect of inducible NO synthase (NOS2)-derived NO on the activity of GRK2 in the context of septic cardiac dysfunction. C57BL/6 mice were submitted to severe septic injury by cecal ligation and puncture (CLP). Heart function was assessed by isolated and perfused heart, echocardiography, and β-adrenergic receptor binding. GRK2 was determined by immunofluorescence and Western blot analysis in the heart and isolated cardiac myocytes. Sepsis increased NOS2 expression in the heart, increased plasma nitrite + nitrate levels, and reduced isoproterenol-induced isolated ventricle contraction, whole heart tension development, and β-adrenergic receptor density. Treatment with 1400W or with GRK2 inhibitor prevented CLP-induced cardiac hyporesponsiveness 12 and 24 h after CLP. Increased labeling of total and phosphorylated GRK2 was detected in hearts after CLP. With treatment of 1400W or in hearts taken from septic NOS2 knockout mice, the activation of GRK2 was reduced. 1400W or GRK2 inhibitor reduced mortality, improved echocardiographic cardiac parameters, and prevented organ damage. Therefore, during sepsis, NOS2-derived NO increases GRK2, which leads to a reduction in β-adrenergic receptor density, contributing to the heart dysfunction. Isolated cardiac myocyte data indicate that NO acts through the soluble guanylyl cyclase/cGMP/PKG pathway. GRK2 inhibition may be a potential therapeutic target in sepsis-induced cardiac dysfunction.NEW & NOTEWORTHY The main novelty presented here is to show that septic shock induces cardiac hyporesponsiveness to isoproterenol by a mechanism dependent on nitric oxide and mediated by G protein-coupled receptor kinase isoform 2. Therefore

  11. G protein-coupled receptor kinase 2 positively regulates epithelial cell migration

    NARCIS (Netherlands)

    Penela, Petronila; Ribas, Catalina; Aymerich, Ivette; Eijkelkamp, Niels; Barreiro, Olga; Heijnen, Cobi J.; Kavelaars, Annemieke; Sanchez-Madrid, Francisco; Mayor, Federico

    2008-01-01

    Cell migration requires integration of signals arising from both the extracellular matrix and messengers acting through G protein-coupled receptors (GPCRs). We find that increased levels of G protein-coupled receptor kinase 2 (GRK2), a key player in GPCR regulation, poteniate migration of epithelial

  12. Exogenous carbon monoxide inhibits neutrophil infiltration in LPS-induced sepsis by interfering with FPR1 via p38 MAPK but not GRK2

    Science.gov (United States)

    Wang, Xu; Qin, Weiting; Song, Mingming; Zhang, Yisen; Sun, Bingwei

    2016-01-01

    Excessive neutrophil infiltration in vital organs is life-threatening to patients who suffer from sepsis. We identified a critical role of exogenous carbon monoxide (CO) in the inhibition of neutrophil infiltration during lipopolysaccharide (LPS)-induced sepsis. CO delivered from carbon monoxide-releasing molecule 2 (CORM-2) dramatically increased the survival rate of C57BL/6 mice subjected to LPS in vivo. CORM-2 significantly suppressed neutrophil infiltration in liver and lung as well as markers of inflammatory responses. Affymetrix GeneChip array analysis revealed that the increased expression of chemoattractant receptor formyl peptide receptor 1 (FPR1) may contribute to the excessive neutrophil infiltration. The under agarose migration assay demonstrated that LPS stimulation promoted migration to the ligand of FPR1, N-Formyl-Met-Leu-Phe (fMLP) but that CORM-2 treatment inhibited this promotion. Further studies demonstrated that CORM-2 internalized FPR1 by inhibiting p38 mitogen-activated protein kinase (MAPK) but not G protein-coupled receptor kinase 2 (GRK2), which may explain the inhibitory effect of CORM-2 on LPS-stimulated neutrophils. In summary, our study demonstrates that exogenous CO inhibits sepsis-induced neutrophil infiltration by interfering with FPR1 via p38 MAPK but not GRK2. PMID:27144520

  13. Bacillus bombysepticus α-Toxin Binding to G Protein-Coupled Receptor Kinase 2 Regulates cAMP/PKA Signaling Pathway to Induce Host Death.

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    Ping Lin

    2016-03-01

    Full Text Available Bacterial pathogens and their toxins target host receptors, leading to aberrant behavior or host death by changing signaling events through subversion of host intracellular cAMP level. This is an efficient and widespread mechanism of microbial pathogenesis. Previous studies describe toxins that increase cAMP in host cells, resulting in death through G protein-coupled receptor (GPCR signaling pathways by influencing adenylyl cyclase or G protein activity. G protein-coupled receptor kinase 2 (GRK2 has a central role in regulation of GPCR desensitization. However, little information is available about the pathogenic mechanisms of toxins associated with GRK2. Here, we reported a new bacterial toxin-Bacillus bombysepticus (Bb α-toxin that was lethal to host. We showed that Bb α-toxin interacted with BmGRK2. The data demonstrated that Bb α-toxin directly bound to BmGRK2 to promote death by affecting GPCR signaling pathways. This mechanism involved stimulation of Gαs, increase level of cAMP and activation of protein kinase A (PKA. Activated cAMP/PKA signal transduction altered downstream effectors that affected homeostasis and fundamental biological processes, disturbing the structural and functional integrity of cells, resulting in death. Preventing cAMP/PKA signaling transduction by inhibitions (NF449 or H-89 substantially reduced the pathogenicity of Bb α-toxin. The discovery of a toxin-induced host death specifically linked to GRK2 mediated signaling pathway suggested a new model for bacterial toxin action. Characterization of host genes whose expression and function are regulated by Bb α-toxin and GRK2 will offer a deeper understanding of the pathogenesis of infectious diseases caused by pathogens that elevate cAMP.

  14. Effects of the dopamine D2 allosteric modulator, PAOPA, on the expression of GRK2, arrestin-3, ERK1/2, and on receptor internalization.

    Directory of Open Access Journals (Sweden)

    Dipannita Basu

    Full Text Available The activity of G protein-coupled receptors (GPCRs is intricately regulated by a range of intracellular proteins, including G protein-coupled kinases (GRKs and arrestins. Understanding the effects of ligands on these signaling pathways could provide insights into disease pathophysiologies and treatment. The dopamine D2 receptor is a GPCR strongly implicated in the pathophysiology of a range of neurological and neuropsychiatric disorders, particularly schizophrenia. Previous studies from our lab have shown the preclinical efficacy of a novel allosteric drug, 3(R-[(2(S-pyrrolidinylcarbonylamino]-2-oxo-1-pyrrolidineacetamide (PAOPA, in attenuating schizophrenia-like behavioural abnormalities in rodent models of the disease. As an allosteric modulator, PAOPA binds to a site on the D2 receptor, which is distinct from the endogenous ligand-binding site, in order to modulate the binding of the D2 receptor ligand, dopamine. The exact signaling pathways affected by this allosteric modulator are currently unknown. The objectives of this study were to decipher the in vivo effects, in rats, of chronic PAOPA administration on D2 receptor regulatory and downstream molecules, including GRK2, arrestin-3 and extracellular receptor kinase (ERK 1/2. Additionally, an in vitro cellular model was also used to study PAOPA's effects on D2 receptor internalization. Results from western immunoblots showed that chronic PAOPA treatment increased the striatal expression of GRK2 by 41%, arrestin-3 by 34%, phospho-ERK1 by 51% and phospho-ERK2 by 36%. Results also showed that the addition of PAOPA to agonist treatment in cells increased D2 receptor internalization by 33%. This study provides the foundational evidence of putative signaling pathways, and changes in receptor localization, affected by treatment with PAOPA. It improves our understanding on the diverse mechanisms of action of allosteric modulators, while advancing PAOPA's development into a novel drug for the

  15. Structure of Human G Protein-Coupled Receptor Kinase 2 in Complex with the Kinase Inhibitor Balanol

    Energy Technology Data Exchange (ETDEWEB)

    Tesmer, John J.G.; Tesmer, Valerie M.; Lodowski, David T.; Steinhagen, Henning; Huber, Jochen (Sanofi); (Michigan); (Texas)

    2010-07-19

    G protein-coupled receptor kinase 2 (GRK2) is a pharmaceutical target for the treatment of cardiovascular diseases such as congestive heart failure, myocardial infarction, and hypertension. To better understand how nanomolar inhibition and selectivity for GRK2 might be achieved, we have determined crystal structures of human GRK2 in complex with G{beta}{gamma} in the presence and absence of the AGC kinase inhibitor balanol. The selectivity of balanol among human GRKs is assessed.

  16. Regulation of EGF-induced ERK/MAPK Activation and EGFR Internalization by G Protein-coupled Receptor Kinase 2

    Institute of Scientific and Technical Information of China (English)

    Jingxia GAO; Jiali LI; Lan MA

    2005-01-01

    G protein-coupled receptor kinases (GRKs) mediate agonist-induced phosphorylation and desensitization of various G protein-coupled receptors (GPCRs). We investigate the role of GRK2 on epidermal growth factor (EGF) receptor signaling, including EGF-induced extracellular signal-regulated kinase and mitogen-activated protein kinase (ERK/MAPK) activation and EGFR internalization. Immunoprecipitation and immunofluorescence experiments show that EGF stimulates GRK2 binding to EGFR complex and GRK2 translocating from cytoplasm to the plasma membrane in human embryonic kidney 293 cells. Western blotting assay shows that EGF-induced ERK/MAPK phosphorylation increases 1.9-fold, 1.1-fold and 1.5-fold (P<0.05) at time point 30, 60 and 120 min, respectively when the cells were transfected with GRK2,suggesting the regulatory role of GRK2 on EGF-induced ERK/MAPK activation. Flow cytometry experiments show that GRK2 overexpression has no effect on EGF-induced EGFR internalization, however, it increases agonist-induced G protein-coupled δ opioid receptor internalization by approximately 40% (P<0.01). Overall,these data suggest that GRK2 has a regulatory role in EGF-induced ERK/MAPK activation, and that the mechanisms underlying the modulatory role of GRK2 in EGFR and GPCR signaling pathways are somewhat different at least in receptor internalization.

  17. Dysfunction of G Protein-Coupled Receptor Kinases in Alzheimer’'s Disease

    Directory of Open Access Journals (Sweden)

    William Z. Suo

    2010-01-01

    Full Text Available Although mutations and variations of several genes have been identified to be involved in Alzheimer's disease (AD, the efforts towards understanding the pathogenic mechanisms of the disease still have a long journey to go. One such effort is to identify the signal transduction deficits, for which previous studies have suggested that the central problems appear to be at the interface between G proteins and their coupled receptors. G protein-coupled receptor kinases (GRKs are a small family of serine/threonine protein kinases primarily acting at the “receptor-G protein interface””. Recent studies have indicated the possible involvement of GRK, primarily GRK2 and GRK5, dysfunction in the pathogenesis of AD. It seems that mild, soluble, β-amyloid accumulation can lead to a reduced membrane (functional and an elevated cytosolic GRK2/5. The increased cytosolic GRK2 appears to be colocalized with damaged mitochondria and neurofibrillary tangles. Moreover, the total levels of GRK2, not only in the brain, but also in peripheral blood samples, are increased in a manner inversely correlated with the patient's cognitive levels. The deficiency of GRK5, on the other hand, impairs presynaptic M2 autoreceptor desensitization, which leads to a reduced acetylcholine release, axonal/synaptic degenerative changes, and associated amnestic, mild cognitive impairment. It also promotes an evil cycle to further increase Beta-amyloid accumulation and exaggerates brain inflammation, possibly even the basal forebrain cholinergic degeneration. Therefore, continuous efforts in this direction are necessary before translating the knowledge to any therapeutic strategies.

  18. Distinct cellular and subcellular distributions of G protein-coupled receptor kinase and arrestin isoforms in the striatum.

    Directory of Open Access Journals (Sweden)

    Evgeny Bychkov

    Full Text Available G protein-coupled receptor kinases (GRKs and arrestins mediate desensitization of G protein-coupled receptors (GPCR. Arrestins also mediate G protein-independent signaling via GPCRs. Since GRK and arrestins demonstrate no strict receptor specificity, their functions in the brain may depend on their cellular complement, expression level, and subcellular targeting. However, cellular expression and subcellular distribution of GRKs and arrestins in the brain is largely unknown. We show that GRK isoforms GRK2 and GRK5 are similarly expressed in direct and indirect pathway neurons in the rat striatum. Arrestin-2 and arrestin-3 are also expressed in neurons of both pathways. Cholinergic interneurons are enriched in GRK2, arrestin-3, and GRK5. Parvalbumin-positive interneurons express more of GRK2 and less of arrestin-2 than medium spiny neurons. The GRK5 subcellular distribution in the human striatal neurons is altered by its phosphorylation: unphosphorylated enzyme preferentially localizes to synaptic membranes, whereas phosphorylated GRK5 is found in plasma membrane and cytosolic fractions. Both GRK isoforms are abundant in the nucleus of human striatal neurons, whereas the proportion of both arrestins in the nucleus was equally low. However, overall higher expression of arrestin-2 yields high enough concentration in the nucleus to mediate nuclear functions. These data suggest cell type- and subcellular compartment-dependent differences in GRK/arrestin-mediated desensitization and signaling.

  19. Increased Atrial β-Adrenergic Receptors and GRK-2 Gene Expression Can Play a Fundamental Role in Heart Failure After Repair of Congenital Heart Disease with Cardiopulmonary Bypass.

    Science.gov (United States)

    Oliveira, Marcela Silva; Carmona, Fabio; Vicente, Walter V A; Manso, Paulo H; Mata, Karina M; Celes, Mara Rúbia; Campos, Erica C; Ramos, Simone G

    2017-04-01

    Surgeries to correct congenital heart diseases are increasing in Brazil and worldwide. However, even with the advances in surgical techniques and perfusion, some cases, especially the more complex ones, can develop heart failure and death. A retrospective study of patients who underwent surgery for correction of congenital heart diseases with cardiopulmonary bypass (CPB) in a university tertiary-care hospital that died, showed infarction in different stages of evolution and scattered microcalcifications in the myocardium, even without coronary obstruction. CPB is a process routinely used during cardiac surgery for congenital heart disease. However, CPB has been related to increased endogenous catecholamines that can lead to major injuries in cardiomyocytes. The mechanisms involved are not completely understood. The aim of this study was to evaluate the alterations induced in the β-adrenergic receptors and GRK-2 present in atrial cardiomyocytes of infants with congenital heart disease undergoing surgical repair with CPB and correlate the alterations with functional and biochemical markers of ischemia/myocardial injury. The study consisted of right atrial biopsies of infants undergoing surgical correction in HC-FMRPUSP. Thirty-three cases were selected. Atrial biopsies were obtained at the beginning of CPB (group G1) and at the end of CPB (group G2). Real-time PCR, Western blotting, and immunofluorescence analysis were conducted to evaluate the expression of β1, β2-adrenergic receptors, and GRK-2 in atrial myocardium. Cardiac function was evaluated by echocardiography and biochemical analysis (N-terminal pro-brain natriuretic peptide (NT-ProBNP), lactate, and cardiac troponin I). We observed an increase in serum lactate, NT-proBNP, and troponin I at the end of CPB indicating tissue hypoxia/ischemia. Even without major clinical consequences in cardiac function, these alterations were followed by a significant increase in gene expression of β1 and β2 receptors and

  20. Molecular Mechanism for Inhibition of G Protein-Coupled Receptor Kinase 2 by a Selective RNA Aptamer

    Energy Technology Data Exchange (ETDEWEB)

    Tesmer, Valerie M.; Lennarz, Sabine; Mayer, Günter; Tesmer, John J.G. (Bonn); (Michigan)

    2012-08-31

    Cardiovascular homeostasis is maintained in part by the rapid desensitization of activated heptahelical receptors that have been phosphorylated by G protein-coupled receptor kinase 2 (GRK2). However, during chronic heart failure GRK2 is upregulated and believed to contribute to disease progression. We have determined crystallographic structures of GRK2 bound to an RNA aptamer that potently and selectively inhibits kinase activity. Key to the mechanism of inhibition is the positioning of an adenine nucleotide into the ATP-binding pocket and interactions with the basic {alpha}F-{alpha}G loop region of the GRK2 kinase domain. Constraints imposed on the RNA by the terminal stem of the aptamer also play a role. These results highlight how a high-affinity aptamer can be used to selectively trap a novel conformational state of a protein kinase.

  1. Molecular Mechanism of Selectivity among G Protein-Coupled Receptor Kinase 2 Inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Thal, David M.; Yeow, Raymond Y.; Schoenau, Christian; Huber, Jochen; Tesmer, John J.G. (Sanofi); (Michigan)

    2012-07-11

    G protein-coupled receptors (GPCRs) are key regulators of cell physiology and control processes ranging from glucose homeostasis to contractility of the heart. A major mechanism for the desensitization of activated GPCRs is their phosphorylation by GPCR kinases (GRKs). Overexpression of GRK2 is strongly linked to heart failure, and GRK2 has long been considered a pharmaceutical target for the treatment of cardiovascular disease. Several lead compounds developed by Takeda Pharmaceuticals show high selectivity for GRK2 and therapeutic potential for the treatment of heart failure. To understand how these drugs achieve their selectivity, we determined crystal structures of the bovine GRK2-G{beta}{gamma} complex in the presence of two of these inhibitors. Comparison with the apoGRK2-G{beta}{gamma} structure demonstrates that the compounds bind in the kinase active site in a manner similar to that of the AGC kinase inhibitor balanol. Both balanol and the Takeda compounds induce a slight closure of the kinase domain, the degree of which correlates with the potencies of the inhibitors. Based on our crystal structures and homology modeling, we identified five amino acids surrounding the inhibitor binding site that we hypothesized could contribute to inhibitor selectivity. However, our results indicate that these residues are not major determinants of selectivity among GRK subfamilies. Rather, selectivity is achieved by the stabilization of a unique inactive conformation of the GRK2 kinase domain.

  2. G protein-coupled receptor kinase 2 promotes flaviviridae entry and replication.

    Directory of Open Access Journals (Sweden)

    Caroline Le Sommer

    Full Text Available Flaviviruses cause a wide range of severe diseases ranging from encephalitis to hemorrhagic fever. Discovery of host factors that regulate the fate of flaviviruses in infected cells could provide insight into the molecular mechanisms of infection and therefore facilitate the development of anti-flaviviral drugs. We performed genome-scale siRNA screens to discover human host factors required for yellow fever virus (YFV propagation. Using a 2 × 2 siRNA pool screening format and a duplicate of the screen, we identified a high confidence list of YFV host factors. To find commonalities between flaviviruses, these candidates were compared to host factors previously identified for West Nile virus (WNV and dengue virus (DENV. This comparison highlighted a potential requirement for the G protein-coupled receptor kinase family, GRKs, for flaviviral infection. The YFV host candidate GRK2 (also known as ADRBK1 was validated both in siRNA-mediated knockdown HuH-7 cells and in GRK(-/- mouse embryonic fibroblasts. Additionally, we showed that GRK2 was required for efficient propagation of DENV and Hepatitis C virus (HCV indicating that GRK2 requirement is conserved throughout the Flaviviridae. Finally, we found that GRK2 participates in multiple distinct steps of the flavivirus life cycle by promoting both entry and RNA synthesis. Together, our findings identified GRK2 as a novel regulator of flavivirus infection and suggest that inhibition of GRK2 function may constitute a new approach for treatment of flavivirus associated diseases.

  3. G protein-coupled receptor kinase 2 promotes flaviviridae entry and replication.

    Science.gov (United States)

    Le Sommer, Caroline; Barrows, Nicholas J; Bradrick, Shelton S; Pearson, James L; Garcia-Blanco, Mariano A

    2012-01-01

    Flaviviruses cause a wide range of severe diseases ranging from encephalitis to hemorrhagic fever. Discovery of host factors that regulate the fate of flaviviruses in infected cells could provide insight into the molecular mechanisms of infection and therefore facilitate the development of anti-flaviviral drugs. We performed genome-scale siRNA screens to discover human host factors required for yellow fever virus (YFV) propagation. Using a 2 × 2 siRNA pool screening format and a duplicate of the screen, we identified a high confidence list of YFV host factors. To find commonalities between flaviviruses, these candidates were compared to host factors previously identified for West Nile virus (WNV) and dengue virus (DENV). This comparison highlighted a potential requirement for the G protein-coupled receptor kinase family, GRKs, for flaviviral infection. The YFV host candidate GRK2 (also known as ADRBK1) was validated both in siRNA-mediated knockdown HuH-7 cells and in GRK(-/-) mouse embryonic fibroblasts. Additionally, we showed that GRK2 was required for efficient propagation of DENV and Hepatitis C virus (HCV) indicating that GRK2 requirement is conserved throughout the Flaviviridae. Finally, we found that GRK2 participates in multiple distinct steps of the flavivirus life cycle by promoting both entry and RNA synthesis. Together, our findings identified GRK2 as a novel regulator of flavivirus infection and suggest that inhibition of GRK2 function may constitute a new approach for treatment of flavivirus associated diseases.

  4. Paroxetine Is a Direct Inhibitor of G Protein-Coupled Receptor Kinase 2 and Increases Myocardial Contractility

    Energy Technology Data Exchange (ETDEWEB)

    Thal, David M. [Univ. of Michigan, Ann Arbor, MI (United States); Homan, Kristoff T. [Univ. of Michigan, Ann Arbor, MI (United States); Chen, Jun [Univ. of New Mexico Health Sciences Center, Albuquerque, NM (United States); Wu, Emily K. [Univ. of Michigan, Ann Arbor, MI (United States); Hinkle, Patricia M. [Univ. of Rochester Medical Center, Rochester, NY (United States); Huang, Z. Maggie [Temple Univ. School of Medicine, Philadelphia, Pennsylvania (United States); Chuprun, J. Kurt [Temple Univ. School of Medicine, Philadelphia, Pennsylvania (United States); Song, Jianliang [Temple Univ. School of Medicine, Philadelphia, Pennsylvania (United States); Gao, Erhe [Temple Univ. School of Medicine, Philadelphia, Pennsylvania (United States); Cheung, Joseph Y. [Temple Univ. School of Medicine, Philadelphia, Pennsylvania (United States); Sklar, Larry A. [Univ. of New Mexico Health Sciences Center, Albuquerque, NM (United States); Koch, Walter J. [Temple Univ. School of Medicine, Philadelphia, Pennsylvania (United States); Tesmer, John J.G. [Univ. of Michigan, Ann Arbor, MI (United States)

    2012-08-10

    G protein-coupled receptor kinase 2 (GRK2) is a well-established therapeutic target for the treatment of heart failure. In this paper we identify the selective serotonin reuptake inhibitor (SSRI) paroxetine as a selective inhibitor of GRK2 activity both in vitro and in living cells. In the crystal structure of the GRK2·paroxetine–Gβγ complex, paroxetine binds in the active site of GRK2 and stabilizes the kinase domain in a novel conformation in which a unique regulatory loop forms part of the ligand binding site. Isolated cardiomyocytes show increased isoproterenol-induced shortening and contraction amplitude in the presence of paroxetine, and pretreatment of mice with paroxetine before isoproterenol significantly increases left ventricular inotropic reserve in vivo with no significant effect on heart rate. Neither is observed in the presence of the SSRI fluoxetine. Our structural and functional results validate a widely available drug as a selective chemical probe for GRK2 and represent a starting point for the rational design of more potent and specific GRK2 inhibitors.

  5. Ca2+-dependent inhibition of G protein-coupled receptor kinase 2 by calmodulin.

    Science.gov (United States)

    Haga, K; Tsuga, H; Haga, T

    1997-02-11

    Agonist- or light-dependent phosphorylation of muscarinic acetylcholine receptor m2 subtypes (m2 receptors) or rhodopsin by G protein-coupled receptor kinase 2 (GRK2) was found to be inhibited by calmodulin in a Ca2+-dependent manner. The phosphorylation was fully inhibited in the absence of G protein betagamma subunits and partially inhibited in the presence of betagamma subunits. The dose-response curve for stimulation by betagamma subunits of the m2 and rhodopsin phosphorylation was shifted to the higher concentration of betagamma subunits by addition of Ca2+-calmodulin. The phosphorylation by GRK2 of a glutathione S-transferase fusion protein containing a peptide corresponding to the central part of the third intracellular loop of m2 receptors (I3-GST) was not affected by Ca2+-calmodulin in the presence or absence of betagamma subunits, but the agonist-dependent stimulation of I3-GST phosphorylation by an I3-deleted m2 receptor mutant in the presence of betagamma subunits was suppressed by Ca2+-calmodulin. These results indicate that Ca2+-calmodulin does not directly interact with the catalytic site of GRK2 but inhibits the kinase activity of GRK2 by interfering with the activation of GRK2 by agonist-bound m2 receptors and G protein betagamma subunits. In agreement with the assumption that GRK2 activity is suppressed by the increase in intracellular Ca2+, the sequestration of m2 receptors expressed in Chinese hamster ovary cells was found to be attenuated by the treatment with a Ca2+ ionophore, A23187.

  6. Characterization of G-protein coupled receptor kinase interaction with the neurokinin-1 receptor using bioluminescence resonance energy transfer

    DEFF Research Database (Denmark)

    Jorgensen, Rasmus; Holliday, Nicholas D; Hansen, Jakob L

    2007-01-01

    To analyze the interaction between the neurokinin-1 (NK-1) receptor and G-protein coupled receptor kinases (GRKs), we performed bioluminescence resonance energy transfer(2) (BRET(2)) measurements between the family A NK-1 receptor and GRK2 and GRK5 as well as their respective kinase-inactive muta......To analyze the interaction between the neurokinin-1 (NK-1) receptor and G-protein coupled receptor kinases (GRKs), we performed bioluminescence resonance energy transfer(2) (BRET(2)) measurements between the family A NK-1 receptor and GRK2 and GRK5 as well as their respective kinase...

  7. G protein-coupled receptor kinase 2 and beta-arrestins are recruited to FSH receptor in stimulated rat primary Sertoli cells.

    Science.gov (United States)

    Marion, Sébastien; Kara, Elodie; Crepieux, Pascale; Piketty, Vincent; Martinat, Nadine; Guillou, Florian; Reiter, Eric

    2006-08-01

    FSH-receptor (FSH-R) signaling is regulated by agonist-induced desensitization and internalization. It has been shown, in a variety of overexpression systems, that G protein-coupled receptor kinases (GRKs) phosphorylate the activated FSH-R, promote beta-arrestin recruitment and ultimately lead to internalization. The accuracy of this mechanism has not yet been demonstrated in cells expressing these different molecules at physiological levels. Using sucrose gradient fractionation, we show that FSH induces the recruitment of the endogenous GRK 2 and beta-arrestin 1/2 from the cytoplasm to the plasma membrane of rat primary Sertoli cells. As assessed by ligand binding, the FSH-R was found expressed in the fractions where GRK 2 and beta-arrestins were recruited upon FSH treatment. In addition, the endogenous beta-arrestin 1 was found dephosphorylated in an agonist-dependent manner. Finally, a significant FSH-binding activity was co-immunoprecipitated with the endogenous beta-arrestins from agonist-stimulated but not from untreated Sertoli cell extracts. This FSH-R interaction with beta-arrestins was sustained for up to 30 min. In conclusion, our data strongly suggest that the GRK/beta-arrestin machinery plays a physiologically relevant role in the regulation of the FSH signaling.

  8. Differential requirements of arrestin-3 and clathrin for ligand-dependent and -independent internalization of human G protein-coupled receptor 40.

    Science.gov (United States)

    Qian, Jing; Wu, Chun; Chen, Xiaopan; Li, Xiangmei; Ying, Guoyuan; Jin, Lili; Ma, Qiang; Li, Guo; Shi, Ying; Zhang, Guozheng; Zhou, Naiming

    2014-11-01

    G protein-coupled receptor 40 (GPR40) is believed to be an attractive target to enhance insulin secretion in patients with type 2 diabetes. GPR40 has been found to couple to Gq protein, leading to the activation of phospholipase C and subsequent increases in the intracellular Ca(2+) level. However, the underlying mechanisms that regulate the internalization and desensitization of GPR40 remain to be elucidated. In the present study, a construct of GPR40 fused with enhanced green fluorescent protein (EGFP) at its C-terminus was constructed for direct imaging of the localization and internalization of GPR40 by confocal microscopy. In stably transfected HEK-293 cells, GPR40 receptors underwent rapid agonist-induced internalization and constitutive ligand-independent internalization. Our data demonstrated that the agonist-mediated internalization of GPR40 was significantly blocked by hypertonic sucrose treatment and by siRNA mediated depletion of the heavy chain of clathrin. In contrast, constitutive GPR40 internalization was not affected by hypertonic sucrose or by knock-down of clathrin expression, but it was affected by treatment with methyl-β-cyclodextrin (MβCD) and nystatin. Furthermore, our results using an arrestin-3-EGFP redistribution assay and siRNA-mediated knock-down of arrestin-3 and GRK2 expression revealed that arrestin-3 and GRK2 play an essential role in the regulation of agonist-mediated GPR40 internalization, but are not involved in the regulation of constitutive GPR40 internalization. Additionally, our observation showed that upon activation by agonist, the internalized GPR40 receptors were rapidly recycled back to the plasma membrane via Rab4/Rab5 positive endosomes, whereas the constitutively internalized GPR40 receptors were recycled back to the cell surface through Rab5 positive endosomes. Because FFA receptors exhibit a high level of homology, our observations could be applicable to other members of this family.

  9. Identification and Structure-Function Analysis of Subfamily Selective G Protein-Coupled Receptor Kinase Inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Homan, Kristoff T.; Larimore, Kelly M.; Elkins, Jonathan M.; Szklarz, Marta; Knapp, Stefan; Tesmer, John J.G. [Michigan; (Oxford)

    2015-02-13

    Selective inhibitors of individual subfamilies of G protein-coupled receptor kinases (GRKs) would serve as useful chemical probes as well as leads for therapeutic applications ranging from heart failure to Parkinson’s disease. To identify such inhibitors, differential scanning fluorimetry was used to screen a collection of known protein kinase inhibitors that could increase the melting points of the two most ubiquitously expressed GRKs: GRK2 and GRK5. Enzymatic assays on 14 of the most stabilizing hits revealed that three exhibit nanomolar potency of inhibition for individual GRKs, some of which exhibiting orders of magnitude selectivity. Most of the identified compounds can be clustered into two chemical classes: indazole/dihydropyrimidine-containing compounds that are selective for GRK2 and pyrrolopyrimidine-containing compounds that potently inhibit GRK1 and GRK5 but with more modest selectivity. The two most potent inhibitors representing each class, GSK180736A and GSK2163632A, were cocrystallized with GRK2 and GRK1, and their atomic structures were determined to 2.6 and 1.85 Å spacings, respectively. GSK180736A, developed as a Rho-associated, coiled-coil-containing protein kinase inhibitor, binds to GRK2 in a manner analogous to that of paroxetine, whereas GSK2163632A, developed as an insulin-like growth factor 1 receptor inhibitor, occupies a novel region of the GRK active site cleft that could likely be exploited to achieve more selectivity. However, neither compound inhibits GRKs more potently than their initial targets. This data provides the foundation for future efforts to rationally design even more potent and selective GRK inhibitors.

  10. G Protein–Coupled Receptor Kinase 2 Plays a Relevant Role in Insulin Resistance and Obesity

    Science.gov (United States)

    Garcia-Guerra, Lucia; Nieto-Vazquez, Iria; Vila-Bedmar, Rocio; Jurado-Pueyo, María; Zalba, Guillermo; Díez, Javier; Murga, Cristina; Fernández-Veledo, Sonia; Mayor, Federico; Lorenzo, Margarita

    2010-01-01

    OBJECTIVE Insulin resistance is associated with the pathogenesis of metabolic disorders as type 2 diabetes and obesity. Given the emerging role of signal transduction in these syndromes, we set out to explore the possible role that G protein–coupled receptor kinase 2 (GRK2), first identified as a G protein–coupled receptor regulator, could have as a modulator of insulin responses. RESEARCH DESIGN AND METHODS We analyzed the influence of GRK2 levels in insulin signaling in myoblasts and adipocytes with experimentally increased or silenced levels of GRK2, as well as in GRK2 hemizygous animals expressing 50% lower levels of this kinase in three different models of insulin resistance: tumor necrosis factor-α (TNF-α) infusion, aging, and high-fat diet (HFD). Glucose transport, whole-body glucose and insulin tolerance, the activation status of insulin pathway components, and the circulating levels of important mediators were measured. The development of obesity and adipocyte size with age and HFD was analyzed. RESULTS Altering GRK2 levels markedly modifies insulin-mediated signaling in cultured adipocytes and myocytes. GRK2 levels are increased by ∼2-fold in muscle and adipose tissue in the animal models tested, as well as in lymphocytes from metabolic syndrome patients. In contrast, hemizygous GRK2 mice show enhanced insulin sensitivity and do not develop insulin resistance by TNF-α, aging, or HFD. Furthermore, reduced GRK2 levels induce a lean phenotype and decrease age-related adiposity. CONCLUSIONS Overall, our data identify GRK2 as an important negative regulator of insulin effects, key to the etiopathogenesis of insulin resistance and obesity, which uncovers this protein as a potential therapeutic target in the treatment of these disorders. PMID:20627936

  11. Quantification of Protein Levels in Single Living Cells

    Directory of Open Access Journals (Sweden)

    Chiu-An Lo

    2015-12-01

    Full Text Available Accurate measurement of the amount of specific protein a cell produces is important for investigating basic molecular processes. We have developed a technique that allows for quantitation of protein levels in single cells in vivo. This protein quantitation ratioing (PQR technique uses a genetic tag that produces a stoichiometric ratio of a fluorescent protein reporter and the protein of interest during protein translation. The fluorescence intensity is proportional to the number of molecules produced of the protein of interest and is used to determine the relative amount of protein within the cell. We use PQR to quantify protein expression of different genes using quantitative imaging, electrophysiology, and phenotype. We use genome editing to insert Protein Quantitation Reporters into endogenous genomic loci in three different genomes for quantitation of endogenous protein levels. The PQR technique will allow for a wide range of quantitative experiments examining gene-to-phenotype relationships with greater accuracy.

  12. Yeast prions: Paramutation at the protein level?

    Science.gov (United States)

    Tuite, Mick F

    2015-08-01

    Prions are proteins that have the potential to refold into a novel conformation that templates the conversion of like molecules to the altered infectious form. In the yeast Saccharomyces cerevisiae, trans-generational epigenetic inheritance can be mediated by a number of structurally and functionally diverse prions. Prionogenesis can confer both loss-of-function and gain-of-function properties to the prion protein and this in turn can have a major impact on host phenotype, short-term adaptation and evolution of new traits. Prionogenesis shares a number of properties in common with paramutation and can be considered as a mitotically and meiotically heritable change in protein conformation induced by trans-interactions between homologous proteins.

  13. Competing G protein-coupled receptor kinases balance G protein and β-arrestin signaling.

    Science.gov (United States)

    Heitzler, Domitille; Durand, Guillaume; Gallay, Nathalie; Rizk, Aurélien; Ahn, Seungkirl; Kim, Jihee; Violin, Jonathan D; Dupuy, Laurence; Gauthier, Christophe; Piketty, Vincent; Crépieux, Pascale; Poupon, Anne; Clément, Frédérique; Fages, François; Lefkowitz, Robert J; Reiter, Eric

    2012-06-26

    Seven-transmembrane receptors (7TMRs) are involved in nearly all aspects of chemical communications and represent major drug targets. 7TMRs transmit their signals not only via heterotrimeric G proteins but also through β-arrestins, whose recruitment to the activated receptor is regulated by G protein-coupled receptor kinases (GRKs). In this paper, we combined experimental approaches with computational modeling to decipher the molecular mechanisms as well as the hidden dynamics governing extracellular signal-regulated kinase (ERK) activation by the angiotensin II type 1A receptor (AT(1A)R) in human embryonic kidney (HEK)293 cells. We built an abstracted ordinary differential equations (ODE)-based model that captured the available knowledge and experimental data. We inferred the unknown parameters by simultaneously fitting experimental data generated in both control and perturbed conditions. We demonstrate that, in addition to its well-established function in the desensitization of G-protein activation, GRK2 exerts a strong negative effect on β-arrestin-dependent signaling through its competition with GRK5 and 6 for receptor phosphorylation. Importantly, we experimentally confirmed the validity of this novel GRK2-dependent mechanism in both primary vascular smooth muscle cells naturally expressing the AT(1A)R, and HEK293 cells expressing other 7TMRs.

  14. Distribution of Vibrational Energy Levels of Protein Molecular Chains

    Institute of Scientific and Technical Information of China (English)

    PANG Xiao-Feng; CHEN Xiang-Rong

    2001-01-01

    The distributions of the quantum vibrational energy levels of the protein molecular chain are found by the discretely nonlinear Schrodinger equation appropriate to protein obtained from the Davydov theory. The results calculated by this method are basically consistent with the experimental values. Furthermore, the energy spectra at high excited states have also been obtained for the molecular chain which is helpful in researching the properties of infrared absorption and Raman scattering of the protein molecules.

  15. Effects of dietary protein levels on the growth performance of ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-07-18

    Jul 18, 2008 ... tion in dietary protein level without affecting fish growth ... management, environmental factors and fish size can .... water quality is thus considered suitable for fish ... Specific growth rate follows the same trend, it increases.

  16. The clinical expression of hereditary protein C and protein S deficiency: : a relation to clinical thrombotic risk-factors and to levels of protein C and protein S

    NARCIS (Netherlands)

    Henkens, C. M. A.; van der Meer, J.; Hillege, J. L.; Bom, V. J. J.; Halie, M. R.; van der Schaaf, W.

    We investigated 103 first-degree relatives of 13 unrelated protein C or protein S deficient patients to assess the role of additional thrombotic risk factors and of protein C and protein S levels in the clinical expression of hereditary protein C and protein S deficiency. Fifty-seven relatives were

  17. Genetic and environmental influences of surfactant protein D serum levels

    DEFF Research Database (Denmark)

    Sørensen, Grith Lykke; Hjelmborg, Jacob v. B.; Kyvik, Kirsten Ohm

    2006-01-01

    in the NH(2)-terminal region (Met11Thr) of the mature protein is significantly associated with the serum SP-D levels. A classic twin study was performed on a twin population including 1,476 self-reported healthy adults. The serum SP-D levels increased with male sex, age, and smoking status. The intraclass...

  18. Soy protein isolate molecular level contributions to bulk adhesive properties

    Science.gov (United States)

    Shera, Jeanne Norton

    Increasing environmental awareness and the recognized health hazards of formaldehyde-based resins has prompted a strong demand for environmentally-responsible adhesives for wood composites. Soy protein-based adhesives have been shown to be commercially viable with 90-day shelf stability and composite physical properties comparable to those of commercial formaldehyde-based particleboards. The main research focus is to isolate and characterize the molecular level features in soy protein isolate responsible for providing mechanical properties, storage stability, and water resistance during adhesive formulation, processing, and wood composite fabrication. Commercial composite board will be reviewed to enhance our understanding of the individual components and processes required for particleboard production. The levels of protein structure will be defined and an overview of current bio-based technology will be presented. In the process, the logic for utilizing soy protein as a sole binder in the adhesive will be reinforced. Variables such as adhesive components, pH, divalent ions, blend aging, protein molecular weight, formulation solids content, and soy protein functionalization will relate the bulk properties of soy protein adhesives to the molecular configuration of the soybean protein. This work has demonstrated that when intermolecular beta-sheet interactions and protein long-range order is disrupted, viscosity and mechanical properties decrease. Storage stability can be maintained through the stabilization of intermolecular beta-sheet interactions. When molecular weight is reduced through enzymatic digestion, long-range order is disrupted and viscosity and mechanical properties decrease accordingly. Processibility and physical properties must be balanced to increase solids while maintaining low viscosity, desirable mechanical properties, and adequate storage stability. The structure of the soybean protein must be related to the particleboard bulk mechanical

  19. Plasma protein carbonyl levels and breast cancer risk.

    Science.gov (United States)

    Rossner, Pavel; Terry, Mary Beth; Gammon, Marilie D; Agrawal, Meenakshi; Zhang, Fang Fang; Ferris, Jennifer S; Teitelbaum, Susan L; Eng, Sybil M; Gaudet, Mia M; Neugut, Alfred I; Santella, Regina M

    2007-01-01

    To study the role of oxidative stress in breast cancer risk, we analysed plasma levels of protein carbonyls in 1050 cases and 1107 controls. We found a statistically significant trend in breast cancer risk in relation to increasing quartiles of plasma protein carbonyl levels (OR = 1.2, 95% CI = 0.9-1.5; OR = 1.5, 95% CI = 1.2-2.0; OR = 1.6, 95% CI = 1.2-2.1, for the 2(nd), 3(rd) and 4(th) quartile relative to the lowest quartile, respectively, P for trend = 0.0001). The increase in risk was similar for younger ( or = 15 grams/day for 4(th) quartile versus lowest quartile OR = 2.3, 95% CI = 1.1-4.7), and hormone replacement therapy use (HRT, OR = 2.6, 95% CI = 1.6-4.4 for 4(th) quartile versus lowest quartile). The multiplicative interaction terms were statistically significant only for physical activity and HRT. The positive association between plasma protein carbonyl levels and breast cancer risk was also observed when the analysis was restricted to women who had not received chemotherapy or radiation therapy prior to blood collection. Among controls, oxidized protein levels significantly increased with cigarette smoking and higher fruit and vegetable consumption, and decreased with alcohol consumption >30 grams per day. Women with higher levels of plasma protein carbonyl and urinary 15F(2t)-isoprostane had an 80% increase in breast cancer risk (OR = 1.8, 95% CI = 1.2-2.6) compared to women with levels below the median for both markers of oxidative stress. In summary, our results suggest that increased plasma protein carbonyl levels may be associated with breast cancer risk.

  20. Multi-level machine learning prediction of protein-protein interactions in Saccharomyces cerevisiae.

    Science.gov (United States)

    Zubek, Julian; Tatjewski, Marcin; Boniecki, Adam; Mnich, Maciej; Basu, Subhadip; Plewczynski, Dariusz

    2015-01-01

    Accurate identification of protein-protein interactions (PPI) is the key step in understanding proteins' biological functions, which are typically context-dependent. Many existing PPI predictors rely on aggregated features from protein sequences, however only a few methods exploit local information about specific residue contacts. In this work we present a two-stage machine learning approach for prediction of protein-protein interactions. We start with the carefully filtered data on protein complexes available for Saccharomyces cerevisiae in the Protein Data Bank (PDB) database. First, we build linear descriptions of interacting and non-interacting sequence segment pairs based on their inter-residue distances. Secondly, we train machine learning classifiers to predict binary segment interactions for any two short sequence fragments. The final prediction of the protein-protein interaction is done using the 2D matrix representation of all-against-all possible interacting sequence segments of both analysed proteins. The level-I predictor achieves 0.88 AUC for micro-scale, i.e., residue-level prediction. The level-II predictor improves the results further by a more complex learning paradigm. We perform 30-fold macro-scale, i.e., protein-level cross-validation experiment. The level-II predictor using PSIPRED-predicted secondary structure reaches 0.70 precision, 0.68 recall, and 0.70 AUC, whereas other popular methods provide results below 0.6 threshold (recall, precision, AUC). Our results demonstrate that multi-scale sequence features aggregation procedure is able to improve the machine learning results by more than 10% as compared to other sequence representations. Prepared datasets and source code for our experimental pipeline are freely available for download from: http://zubekj.github.io/mlppi/ (open source Python implementation, OS independent).

  1. Secreted Proteins Defy the Expression Level-Evolutionary Rate Anticorrelation.

    Science.gov (United States)

    Feyertag, Felix; Berninsone, Patricia M; Alvarez-Ponce, David

    2017-03-01

    The rates of evolution of the proteins of any organism vary across orders of magnitude. A primary factor influencing rates of protein evolution is expression. A strong negative correlation between expression levels and evolutionary rates (the so-called E-R anticorrelation) has been observed in virtually all studied organisms. This effect is currently attributed to the abundance-dependent fitness costs of misfolding and unspecific protein-protein interactions, among other factors. Secreted proteins are folded in the endoplasmic reticulum, a compartment where chaperones, folding catalysts, and stringent quality control mechanisms promote their correct folding and may reduce the fitness costs of misfolding. In addition, confinement of secreted proteins to the extracellular space may reduce misinteractions and their deleterious effects. We hypothesize that each of these factors (the secretory pathway quality control and extracellular location) may reduce the strength of the E-R anticorrelation. Indeed, here we show that among human proteins that are secreted to the extracellular space, rates of evolution do not correlate with protein abundances. This trend is robust to controlling for several potentially confounding factors and is also observed when analyzing protein abundance data for 6 human tissues. In addition, analysis of mRNA abundance data for 32 human tissues shows that the E-R correlation is always less negative, and sometimes nonsignificant, in secreted proteins. Similar observations were made in Caenorhabditis elegans and in Escherichia coli, and to a lesser extent in Drosophila melanogaster, Saccharomyces cerevisiae and Arabidopsis thaliana. Our observations contribute to understand the causes of the E-R anticorrelation. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Membrane orientation and binding determinants of G protein-coupled receptor kinase 5 as assessed by combined vibrational spectroscopic studies.

    Directory of Open Access Journals (Sweden)

    Pei Yang

    Full Text Available G-protein coupled receptors (GPCRs are integral membrane proteins involved in a wide variety of biological processes in eukaryotic cells, and are targeted by a large fraction of marketed drugs. GPCR kinases (GRKs play important roles in feedback regulation of GPCRs, such as of β-adrenergic receptors in the heart, where GRK2 and GRK5 are the major isoforms expressed. Membrane targeting is essential for GRK function in cells. Whereas GRK2 is recruited to the membrane by heterotrimeric Gβγ subunits, the mechanism of membrane binding by GRK5 is not fully understood. It has been proposed that GRK5 is constitutively associated with membranes through elements located at its N-terminus, its C-terminus, or both. The membrane orientation of GRK5 is also a matter of speculation. In this work, we combined sum frequency generation (SFG vibrational spectroscopy and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR to help determine the membrane orientation of GRK5 and a C-terminally truncated mutant (GRK51-531 on membrane lipid bilayers. It was found that GRK5 and GRK51-531 adopt a similar orientation on model cell membranes in the presence of PIP2 that is similar to that predicted for GRK2 in prior studies. Mutation of the N-terminal membrane binding site of GRK5 did not eliminate membrane binding, but prevented observation of this discrete orientation. The C-terminus of GRK5 does not have substantial impact on either membrane binding or orientation in this model system. Thus, the C-terminus of GRK5 may drive membrane binding in cells via interactions with other proteins at the plasma membrane or bind in an unstructured manner to negatively charged membranes.

  3. THE CLINICAL EXPRESSION OF HEREDITARY PROTEIN-C AND PROTEIN-S DEFICIENCY - A RELATION TO CLINICAL THROMBOTIC RISK-FACTORS AND TO LEVELS OF PROTEIN-C AND PROTEIN-S

    NARCIS (Netherlands)

    HENKENS, CMA; VANDERMEER, J; HILLEGE, JL; BOM, VJJ; HALIE, MR; van der Schaaf, W

    1993-01-01

    We investigated 103 first-degree relatives of 13 unrelated protein C or protein S deficient patients to assess the role of additional thrombotic risk factors and of protein C and protein S levels in the clinical expression of hereditary protein C and protein S deficiency. Fifty-seven relatives were

  4. Low copper and high manganese levels in prion protein plaques

    Science.gov (United States)

    Johnson, Christopher J.; Gilbert, P.U.P.A.; Abrecth, Mike; Baldwin, Katherine L.; Russell, Robin E.; Pedersen, Joel A.; McKenzie, Debbie

    2013-01-01

    Accumulation of aggregates rich in an abnormally folded form of the prion protein characterize the neurodegeneration caused by transmissible spongiform encephalopathies (TSEs). The molecular triggers of plaque formation and neurodegeneration remain unknown, but analyses of TSE-infected brain homogenates and preparations enriched for abnormal prion protein suggest that reduced levels of copper and increased levels of manganese are associated with disease. The objectives of this study were to: (1) assess copper and manganese levels in healthy and TSE-infected Syrian hamster brain homogenates; (2) determine if the distribution of these metals can be mapped in TSE-infected brain tissue using X-ray photoelectron emission microscopy (X-PEEM) with synchrotron radiation; and (3) use X-PEEM to assess the relative amounts of copper and manganese in prion plaques in situ. In agreement with studies of other TSEs and species, we found reduced brain levels of copper and increased levels of manganese associated with disease in our hamster model. We also found that the in situ levels of these metals in brainstem were sufficient to image by X-PEEM. Using immunolabeled prion plaques in directly adjacent tissue sections to identify regions to image by X-PEEM, we found a statistically significant relationship of copper-manganese dysregulation in prion plaques: copper was depleted whereas manganese was enriched. These data provide evidence for prion plaques altering local transition metal distribution in the TSE-infected central nervous system.

  5. UCP2, a mitochondrial protein regulated at multiple levels.

    Science.gov (United States)

    Donadelli, Massimo; Dando, Ilaria; Fiorini, Claudia; Palmieri, Marta

    2014-04-01

    An ever-increasing number of studies highlight the role of uncoupling protein 2 (UCP2) in a broad range of physiological and pathological processes. The knowledge of the molecular mechanisms of UCP2 regulation is becoming fundamental in both the comprehension of UCP2-related physiological events and the identification of novel therapeutic strategies based on UCP2 modulation. The study of UCP2 regulation is a fast-moving field. Recently, several research groups have made a great effort to thoroughly understand the various molecular mechanisms at the basis of UCP2 regulation. In this review, we describe novel findings concerning events that can occur in a concerted manner at various levels: Ucp2 gene mutation (single nucleotide polymorphisms), UCP2 mRNA and protein expression (transcriptional, translational, and protein turn-over regulation), UCP2 proton conductance (ligands and post-transcriptional modifications), and nutritional and pharmacological regulation of UCP2.

  6. Oxidative stress causes renal dopamine D1 receptor dysfunction and hypertension via mechanisms that involve nuclear factor-kappaB and protein kinase C.

    Science.gov (United States)

    Banday, Anees Ahmad; Fazili, Fatima Rizwan; Lokhandwala, Mustafa F

    2007-05-01

    Renal dopamine, via activation of D1 receptors, plays a role in maintaining sodium homeostasis and BP. There exists a defect in renal D1 receptor function in hypertension, diabetes, and aging, conditions that are associated with oxidative stress. However, the exact underlying mechanism of the oxidative stress-mediated impaired D1 receptor signaling and hypertension is not known. The effect of oxidative stress on renal D1 receptor function was investigated in healthy animals. Male Sprague-Dawley rats received tap water (vehicle) and 30 mM L-buthionine sulfoximine (BSO), an oxidant, with and without 1 mM tempol for 2 wk. Compared with vehicle, BSO treatment caused oxidative stress and increase in BP, which was accompanied by defective D1 receptor G-protein coupling and loss of natriuretic response to SKF38393. BSO treatment also increased NF-kappaB nuclear translocation, protein kinase C (PKC) activity and expression, G-protein-coupled receptor kinase-2 (GRK-2) membranous translocation, and D1 receptor serine phosphorylation. In BSO-treated rats' supplementation of tempol decreased oxidative stress, normalized BP, and restored D1 receptor G-protein coupling and natriuretic response to SKF38393. Tempol also normalized NF-kappaB translocation, PKC activity and expression, GRK-2 sequestration, and D1 receptor serine phosphorylation. In conclusion, these results show that oxidative stress activates NF-kappaB, causing an increase in PKC activity, which leads to GRK-2 translocation and subsequent D1 receptor hyper-serine phosphorylation and uncoupling. The functional consequence of this phenomenon was the inability of SKF38393 to inhibit Na/K-ATPase activity and promote sodium excretion, which may have contributed to increase in BP. Tempol reduced oxidative stress and thereby restored D1 receptor function and normalized BP.

  7. Ascertaining effects of nanoscale polymeric interfaces on competitive protein adsorption at the individual protein level.

    Science.gov (United States)

    Song, Sheng; Xie, Tian; Ravensbergen, Kristina; Hahm, Jong-in

    2016-02-14

    With the recent development of biomaterials and biodevices with reduced dimensionality, it is critical to comprehend protein adhesion processes to nanoscale solid surfaces, especially those occurring in a competitive adsorption environment. Complex sequences of adhesion events in competitive adsorption involving multicomponent protein systems have been extensively investigated, but our understanding is still limited primarily to macroscopic adhesion onto chemically simple surfaces. We examine the competitive adsorption behavior from a binary protein mixture containing bovine serum albumin and fibrinogen at the single protein level. We subsequently evaluate a series of adsorption and displacement processes occurring on both the macroscopic homopolymer and nanoscopic diblock copolymer surfaces, while systematically varying the protein concentration and incubation time. We identify the similarities and dissimilarities in competitive protein adsorption behavior between the two polymeric surfaces, the former presenting chemical uniformity at macroscale versus the latter exhibiting periodic nanointerfaces of chemically alternating polymeric segments. We then present our novel experimental finding of a large increase in the nanointerface-engaged residence time of the initially bound proteins and further explain the origin of this phenomenon manifested on nanoscale diblock copolymer surfaces. The outcomes of this study may provide timely insight into nanoscale competitive protein adsorption that is much needed in designing bioimplant and tissue engineering materials. In addition, the fundamental understanding gained from this study can be beneficial for the development of highly miniaturized biodevices and biomaterials fabricated by using nanoscale polymeric materials and interfaces.

  8. Genetic and environmental influences of surfactant protein D serum levels

    DEFF Research Database (Denmark)

    Sorensen, G.L.; Hjelmborg, J.V.; Kyvik, K.O.

    2006-01-01

    defining the constitutional serum level of SP-D and determine the magnitude of the genetic contribution to serum SP-D in the adult population. Recent studies have demonstrated that serum SP-D concentrations in children are genetically determined and that a single nucleotide polymorphism (SNP) located...... in the NH(2)-terminal region (Met11Thr) of the mature protein is significantly associated with the serum SP-D levels. A classic twin study was performed on a twin population including 1,476 self-reported healthy adults. The serum SP-D levels increased with male sex, age, and smoking status. The intraclass...... correlation was significantly higher for monozygotic (MZ) twin pairs than for dizygotic (DZ) twin pairs. Serum SP-D variance was influenced by nonshared environmental effects and additive genetic effects. Multivariate analysis of MZ and DZ covariance matrixes showed significant genetic correlation among serum...

  9. Effect of Crude Protein Levels in Concentrate and Concentrate Levels in Diet on Fermentation

    Directory of Open Access Journals (Sweden)

    Dinh Van Dung

    2014-06-01

    Full Text Available The effect of concentrate mixtures with crude protein (CP levels 10%, 13%, 16%, and 19% and diets with roughage to concentrate ratios 80:20, 60:40, 40:60, and 20:80 (w/w were determined on dry matter (DM and organic matter (OM digestibility, and fermentation metabolites using an in vitro fermentation technique. In vitro fermented attributes were measured after 4, 24, and 48 h of incubation respectively. The digestibility of DM and OM, and total volatile fatty acid (VFA increased whereas pH decreased with the increased amount of concentrate in the diet (p<0.001, however CP levels of concentrate did not have any influence on these attributes. Gas production reduced with increased CP levels, while it increased with increasing concentrate levels. Ammonia nitrogen (NH3-N concentration and microbial CP production increased significantly (p<0.05 by increasing CP levels and with increasing concentrate levels in diet as well, however, no significant difference was found between 16% and 19% CP levels. Therefore, 16% CP in concentrate and increasing proportion of concentrate up to 80% in diet all had improved digestibility of DM and organic matter, and higher microbial protein production, with improved fermentation characteristics.

  10. C-Reactive Protein Levels in the Brugada Syndrome

    Directory of Open Access Journals (Sweden)

    Aimé Bonny

    2011-01-01

    Full Text Available Background. Inflammation in the Brugada syndrome (BrS and its clinical implication have been little studied. Aims. To assess the level of inflammation in BrS patients. Methods. All studied BrS patients underwent blood samples drawn for C-reactive protein (CRP levels at admission, prior to any invasive intervention. Patients with a previous ICD placement were controlled to exclude those with a recent (<14 days shock. We divided subjects into symptomatic (syncope or aborted sudden death and asymptomatic groups. In a multivariable analysis, we adjusted for significant variables (age, CRP ≥ 2 mg/L. Results. Fifty-four subjects were studied (mean age 45 ± 13 years, 49 (91% male. Twenty (37% were symptomatic. Baseline characteristics were similar in both groups. Mean CRP level was 1,4 ± 0,9 mg/L in asymptomatic and 2,4 ± 1,4 mg/L in symptomatic groups (P = .003. In the multivariate model, CRP concentrations ≥ 2 mg/L remained an independent marker for being symptomatic (P = .018; 95% CI: 1.3 to 19.3. Conclusion. Inflammation seems to be more active in symptomatic BrS. C-reactive protein concentrations ≥ 2 mg/L might be associated with the previous symptoms in BrS. The value of inflammation as a risk factor of arrhythmic events in BrS needs to be studied.

  11. C-reactive protein levels in hereditary angioedema.

    Science.gov (United States)

    Hofman, Z L M; Relan, A; Hack, C E

    2014-07-01

    Hereditary angioedema (HAE) patients experience recurrent episodes of angioedema attacks that can be painful, disfiguring and even life-threatening. The disorder results from a mutation in the gene that controls the synthesis of C1-inhibitor (C1INH). C1INH is a major regulator of activation of the contact system. It is often assumed that attacks results from uncontrolled local activation of the contact system with subsequent formation of bradykinin. To evaluate the involvement of inflammatory reactions in HAE, we analysed C-reactive protein (CRP) levels. HAE patients included in a clinical database of recombinant human C1-inhibitor (rhC1INH) studies were evaluated. For the current study we analysed CRP levels when patients were asymptomatic, during a clinical attack and in a follow-up period, and correlated these with the clinical manifestations of the attack. Data from 68 HAE patients were analysed and included CRP levels on 273 occasions. While asymptomatic, 20% of the patients analysed had increased CRP. At the onset of the attack (P = 0·049) and during the next 24 h CRP rose significantly (P = 0·002) in patients with an abdominal location, and post-attack levels were significantly higher in these patients than in patients with attacks at other locations (P = 0·034). In conclusion, CRP levels are elevated in a substantial proportion of asymptomatic HAE patients. Levels of CRP increase significantly during an abdominal attack. These data suggest low-grade systemic inflammatory reactions in HAE patients as well as a triggering event for attacks that starts prior to symptom onset.

  12. Level of supplemental protein does not influence the ruminally undegradable protein value.

    Science.gov (United States)

    Legleiter, L R; Mueller, A M; Kerley, M S

    2005-04-01

    Two experiments were conducted to determine whether elevating the percentage of ruminally undegradable protein (RUP) in the diet would influence the RUP value of the protein feedstuff. A single-effluent, continuous-culture study was designed to test the effect of RUP inclusion rate in the diet on ruminal degradability of the protein. Treatments consisted (DM basis) of a control diet with no supplemental protein, control + 2.5% bloodmeal (BM-L), control + 5% bloodmeal (BM-H), control + 4.45% soybean meal (SBM-L), and control + 8.89% soybean meal (SBM-H). Proteolytic activity and total VFA concentration were not affected (P = 0.73 and P = 0.13) by treatment. Within protein source, dietary RUP value was not affected (P = 0.94) by level of inclusion. When corrected for control diet RUP flow, the RUP value of the blood meal (BM) protein was higher (P = 0.01) than soybean meal (SBM); however, level of supplementation did not affect (P = 0.07) the RUP value of BM or SBM. In Exp. 2, 32 British x Continental crossbred steers (276 +/- 26.3 kg) were fed for 72 d to examine the effects of balancing the AA:energy ratio, using BM as a RUP source, on ADG, G:F, and lean tissue deposition. Diets were formulated to provide increasing levels of arginine, while ruminally degradable protein and energy were held constant. Four dietary treatments provided 0.5, 1, 1.5, and 2x the required amount of arginine, whereas the control diet had no BM included. Daily DMI averaged 7.6 kg/steer and did not differ (P = 0.71) among treatments. Steers gained an average of 1.9 kg/d and average G:F was 0.260, with no differences (P = 0.60 and P = 0.97, respectively) among treatments. There was no difference (P = 0.48) in the change in 12th-rib fat depth during the study; however, change in LM area was affected quadratically as the level of BM increased in the diet, with the greatest increase in LM area occurring in steers fed the 1x and 1.5x required arginine treatments. Balancing the AA:energy ratio did

  13. Influence of level and type of dietary protein, and of level of feeding on feed utilization by rainbow trout.

    Science.gov (United States)

    Cho, C Y; Slinger, S J; Bayley, H S

    1976-11-01

    In the first experiment, diets with 40%, 50% and 60% protein containing herring and soybean meals, and diets with 50% and 60% protein containing herring and soybean protein concentrates were fed at 100%, 85% and 70% of the ad libitum intake. Both increasing the protein level and using the concentrated protein sources resulted in lower gains, and in increases in the amounts of digestible protein and energy consumed for the deposition of protein and energy in the fish carcasses. This difference between the herring and soybean meals, and protein concentrates was not due to differences between their digestibilities. Restriction of feed intake reduced gain, but within these lower levels of gain, the 50% and 60% protein diets have higher final body weights than the 40% protein diet. Restriction of feed intake resulted in carcasses with higher protein and lower fat contents. In the second experiment, two series of diets containing either 40%, 35%, 30% or 25% protein from both herring and soybean meals, or 35%, 30% and 25% protein from herring meal were fed. Reducing the protein level below 40% resulted in lower final body weights, higher feed: gain ratios and carcasses with more protein and less fat. Comparisons of the isonitrogenous pairs of diets showed no differences in final body weights, feed: gain ratios and carcass composition. Reducing the protein level in the diet reduced the amount of protein consumed per unit protein level in the diet reduced the amount of protein consumed per unit protein deposition, but increased the amount of gross energy per unit energy deposited in the carcass.

  14. Honey bee protein atlas at organ-level resolution.

    Science.gov (United States)

    Chan, Queenie W T; Chan, Man Yi; Logan, Michelle; Fang, Yuan; Higo, Heather; Foster, Leonard J

    2013-11-01

    Genome sequencing has provided us with gene lists but cannot tell us where and how their encoded products work together to support life. Complex organisms rely on differential expression of subsets of genes/proteins in organs and tissues, and, in concert, evolved to their present state as they function together to improve an organism's overall reproductive fitness. Proteomics studies of individual organs help us understand their basic functions, but this reductionist approach misses the larger context of the whole organism. This problem could be circumvented if all the organs in an organism were comprehensively studied by the same methodology and analyzed together. Using honey bees (Apis mellifera L.) as a model system, we report here an initial whole proteome of a complex organism, measuring 29 different organ/tissue types among the three honey bee castes: queen, drone, and worker. The data reveal that, e.g., workers have a heightened capacity to deal with environmental toxins and queens have a far more robust pheromone detection system than their nestmates. The data also suggest that workers altruistically sacrifice not only their own reproductive capacity but also their immune potential in favor of their queen. Finally, organ-level resolution of protein expression offers a systematic insight into how organs may have developed.

  15. Probing Protein Channel Dynamics At The Single Molecule Level.

    Science.gov (United States)

    Lee, M. Ann; Dunn, Robert C.

    1997-03-01

    It would be difficult to overstate the importance played by protein ion channels in cellular function. These macromolecular pores allow the passage of ions across the cellular membrane and play indispensable roles in all aspects of neurophysiology. While the patch-clamp technique continues to provide elegant descriptions of the kinetic processes involved in ion channel gating, the associated conformational changes remain a mystery. We are using the spectroscopic capabilities and single molecule fluorescence sensitivity of near-field scanning optical microscopy (NSOM) to probe these dynamics at the single channel level. Using a newly developed cantilevered NSOM probe capable of probing soft biological samples with single molecule fluorescence sensitivity, we have begun mapping the location of single NMDA receptors in intact rat cortical neurons with <100 nm spatial resolution. We will also present recent results exploring the conformational changes accompanying activation of nuclear pore channels located in the nuclear membrane of Xenopus oocytes. Our recent NSOM and AFM measurements on single nuclear pore complexes reveal large conformational changes taking place upon activation, providing rich, new molecular level details of channel function.

  16. Effect of Varying Protein Levels on the Growth of Indian Major Carp Rohu, Labeo rohita (Hamilton

    Directory of Open Access Journals (Sweden)

    M.S. Chari

    2006-01-01

    Full Text Available A forty-nine day feeding experiment conduced to determine protein requirement of fingerlings of Labeo rohita (average weight 1.830.02 g feeding on four formulated diets with varying protein levels (25, 30, 35 and 40% using slaughter house waste as the major protein source. In terms of growth, food conversion ratio, protein efficiency ratio, survival and ratios of protein and lipid deposition in muscle, diet containing 30% protein level revealed a significantly (pLabeo rohita.

  17. Imaging proteins at the single-molecule level

    Science.gov (United States)

    Longchamp, Jean-Nicolas; Rauschenbach, Stephan; Abb, Sabine; Escher, Conrad; Latychevskaia, Tatiana; Kern, Klaus; Fink, Hans-Werner

    2017-01-01

    Imaging single proteins has been a long-standing ambition for advancing various fields in natural science, as for instance structural biology, biophysics, and molecular nanotechnology. In particular, revealing the distinct conformations of an individual protein is of utmost importance. Here, we show the imaging of individual proteins and protein complexes by low-energy electron holography. Samples of individual proteins and protein complexes on ultraclean freestanding graphene were prepared by soft-landing electrospray ion beam deposition, which allows chemical- and conformational-specific selection and gentle deposition. Low-energy electrons do not induce radiation damage, which enables acquiring subnanometer resolution images of individual proteins (cytochrome C and BSA) as well as of protein complexes (hemoglobin), which are not the result of an averaging process. PMID:28087691

  18. A case for protein-level and site-level specificity in glycoproteomic studies of disease.

    Science.gov (United States)

    Schumacher, Katherine N; Dodds, Eric D

    2016-06-01

    Abnormal glycosylation of proteins is known to be either resultant or causative of a variety of diseases. This makes glycoproteins appealing targets as potential biomarkers and focal points of molecular studies on the development and progression of human ailment. To date, a majority of efforts in disease glycoproteomics have tended to center on either determining the concentration of a given glycoprotein, or on profiling the total population of glycans released from a mixture of glycoproteins. While these approaches have demonstrated some diagnostic potential, they are inherently insensitive to the fine molecular detail which distinguishes unique and possibly disease relevant glycoforms of specific proteins. As a consequence, such analyses can be of limited sensitivity, specificity, and accuracy because they do not comprehensively consider the glycosylation status of any particular glycoprotein, or of any particular glycosylation site. Therefore, significant opportunities exist to improve glycoproteomic inquiry into disease by engaging in these studies at the level of individual glycoproteins and their exact loci of glycosylation. In this concise review, the rationale for glycoprotein and glycosylation site specificity is developed in the context of human disease glycoproteomics with an emphasis on N-glycosylation. Recent examples highlighting disease-related perturbations in glycosylation will be presented, including those involving alterations in the overall glycosylation of a specific protein, alterations in the occupancy of a given glycosylation site, and alterations in the compositional heterogeneity of glycans occurring at a given glycosylation site. Each will be discussed with particular emphasis on how protein-specific and site-specific approaches can contribute to improved discrimination between glycoproteomes and glycoproteins associated with healthy and unhealthy states.

  19. Internalization and down-regulation of human muscarinic acetylcholine receptor m2 subtypes. Role of third intracellular m2 loop and G protein-coupled receptor kinase 2.

    Science.gov (United States)

    Tsuga, H; Kameyama, K; Haga, T; Honma, T; Lameh, J; Sadée, W

    1998-02-27

    Internalization and down-regulation of human muscarinic acetylcholine m2 receptors (hm2 receptors) and a hm2 receptor mutant lacking a central part of the third intracellular loop (I3-del m2 receptor) were examined in Chinese hamster ovary (CHO-K1) cells stably expressing these receptors and G protein-coupled receptor kinase 2 (GRK2). Agonist-induced internalization of up to 80-90% of hm2 receptors was demonstrated by measuring loss of [3H]N-methylscopolamine binding sites from the cell surface, and transfer of [3H]quinuclidinyl benzilate binding sites from the plasma membrane into the light-vesicle fractions separated by sucrose density gradient centrifugation. Additionally, translocation of hm2 receptors with endocytic vesicles were visualized by immunofluorescence confocal microscopy. Agonist-induced down-regulation of up to 60-70% of hm2 receptors was demonstrated by determining the loss of [3H]quinuclidinyl benzilate binding sites in the cells. The half-time (t1/2) of internalization and down-regulation in the presence of 10(-4) M carbamylcholine was estimated to be 9.5 min and 2.3 h, respectively. The rates of both internalization and down-regulation of hm2 receptors in the presence of 10(-6) M or lower concentrations of carbamylcholine were markedly increased by coexpression of GRK2. Agonist-induced internalization of I3-del m2 receptors was barely detectable upon incubation of cells for 1 h, but agonist-induced down-regulation of up to 40-50% of I3-del m2 receptors occurred upon incubation with 10(-4) M carbamylcholine for 16 h. However, the rate of down-regulation was lower compared with wild type receptors (t1/2 = 9.9 versus 2.3 h). These results indicate that rapid internalization of hm2 receptors is facilitated by their phosphorylation with GRK2 and does not occur in the absence of the third intracellular loop, but down-regulation of hm2 receptors may occur through both GRK2-facilitating pathway and third intracellular loop-independent pathways.

  20. Gene-specific correlation of RNA and protein levels in human cells and tissues

    DEFF Research Database (Denmark)

    Edfors, Fredrik; Danielsson, Frida; Hallström, Björn M.

    2016-01-01

    to measure, at steady-state conditions, absolute protein copy numbers across human tissues and cell lines and compared these levels with the corresponding mRNA levels using transcriptomics. The study shows that the transcript and protein levels do not correlate well unless a gene-specific RNA-to-protein (RTP...

  1. Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants.

    Science.gov (United States)

    Conley, Andrew J; Joensuu, Jussi J; Richman, Alex; Menassa, Rima

    2011-05-01

    For the past two decades, therapeutic and industrially important proteins have been expressed in plants with varying levels of success. The two major challenges hindering the economical production of plant-made recombinant proteins include inadequate accumulation levels and the lack of efficient purification methods. To address these limitations, several fusion protein strategies have been recently developed to significantly enhance the production yield of plant-made recombinant proteins, while simultaneously assisting in their subsequent purification. Elastin-like polypeptides are thermally responsive biopolymers composed of a repeating pentapeptide 'VPGXG' sequence that are valuable for the purification of recombinant proteins. Hydrophobins are small fungal proteins capable of altering the hydrophobicity of their respective fusion partner, thus enabling efficient purification by surfactant-based aqueous two-phase systems. Zera, a domain of the maize seed storage protein γ-zein, can induce the formation of protein storage bodies, thus facilitating the recovery of fused proteins using density-based separation methods. These three novel protein fusion systems have also been shown to enhance the accumulation of a range of different recombinant proteins, while concurrently inducing the formation of protein bodies. The packing of these fusion proteins into protein bodies may exclude the recombinant protein from normal physiological turnover. Furthermore, these systems allow for quick, simple and inexpensive nonchromatographic purification of the recombinant protein, which can be scaled up to industrial levels of protein production. This review will focus on the similarities and differences of these artificial storage organelles, their biogenesis and their implication for the production of recombinant proteins in plants and their subsequent purification.

  2. Development, characterization, and optimization of protein level in date bars using response surface methodology.

    Science.gov (United States)

    Nadeem, Muhammad; Salim-ur-Rehman; Muhammad Anjum, Faqir; Murtaza, Mian Anjum; Mueen-ud-Din, Ghulam

    2012-01-01

    This project was designed to produce a nourishing date bar with commercial value especially for school going children to meet their body development requirements. Protein level of date bars was optimized using response surface methodology (RSM). Economical and underutilized sources, that is, whey protein concentrate and vetch protein isolates, were explored for protein supplementation. Fourteen date bar treatments were produced using a central composite design (CCD) with 2 variables and 3 levels for each variable. Date bars were then analyzed for nutritional profile. Proximate composition revealed that addition of whey protein concentrate and vetch protein isolates improved the nutritional profile of date bars. Protein level, texture, and taste were considerably improved by incorporating 6.05% whey protein concentrate and 4.35% vetch protein isolates in date bar without affecting any sensory characteristics during storage. Response surface methodology was observed as an economical and effective tool to optimize the ingredient level and to discriminate the interactive effects of independent variables.

  3. EFFECT OF MILK UREA AND PROTEIN LEVELS ON FERTILITY INDICES IN COWS

    Directory of Open Access Journals (Sweden)

    Malgorzata JANKOWSKA

    2011-01-01

    Full Text Available An analysis of the effect of milk urea and protein levels in four consecutive lactations on fertility indices of Blackand- White Polish Holstein-Friesian cows, milk recorded in the Kujawsko-Pomorskie province has been made. Poorer fertility indices were found in first-calf heifers and second lactation cows receiving energy-deficient diets and in older (third and fourth lactation cows receiving excess dietary protein and energy. Best fertility was found in young cows fed excess protein (>3.60% regardless of milk urea levels, and in older cows having lower and optimum levels regardless of protein levels. Cow fertility is differentiated more by milk protein levels than by urea content. Fertility parameters were poorer in first and second lactation cows than in older cows. The coefficients of correlation between milk urea and protein levels and fertility indices were very low, with the only significant differences between protein content vs. calving interval and reproductive rest period.

  4. Nrf2 reduces levels of phosphorylated tau protein by inducing autophagy adaptor protein NDP52

    Science.gov (United States)

    Jo, Chulman; Gundemir, Soner; Pritchard, Susanne; Jin, Youngnam N.; Rahman, Irfan; Johnson, Gail V. W.

    2014-03-01

    Nuclear factor erythroid 2-related factor 2 (Nrf2) is a pivotal transcription factor in the defence against oxidative stress. Here we provide evidence that activation of the Nrf2 pathway reduces the levels of phosphorylated tau by induction of an autophagy adaptor protein NDP52 (also known as CALCOCO2) in neurons. The expression of NDP52, which we show has three antioxidant response elements (AREs) in its promoter region, is strongly induced by Nrf2, and its overexpression facilitates clearance of phosphorylated tau in the presence of an autophagy stimulator. In Nrf2-knockout mice, phosphorylated and sarkosyl-insoluble tau accumulates in the brains concurrent with decreased levels of NDP52. Moreover, NDP52 associates with phosphorylated tau from brain cortical samples of Alzheimer disease cases, and the amount of phosphorylated tau in sarkosyl-insoluble fractions is inversely proportional to that of NDP52. These results suggest that NDP52 plays a key role in autophagy-mediated degradation of phosphorylated tau in vivo.

  5. Plasma levels of soluble endothelial cell protein C receptor in patients with Wegener's granulomatosis

    NARCIS (Netherlands)

    Boomsma, MM; Stearns-Kurosawa, DJ; Stegeman, CA; Raschi, E; Meroni, PL; Kurosawa, S; Tervaert, JWC

    Elevated soluble thrombomodulin (sTM) levels are an accepted marker of endothelial damage. The physiological significance of plasma endothelial protein C receptor (sEPCR) levels is not known. To assess the relevance of this plasma protein in Wegener's granulomatosis (WG), sEPCR levels were measured

  6. Genetic and environmental influences of surfactant protein D serum levels

    DEFF Research Database (Denmark)

    Sorensen, G.L.; Hjelmborg, J.V.; Kyvik, K.O.

    2006-01-01

    The collectin surfactant protein D (SP-D) is an important component of the pulmonary innate immune system, but SP-D is also present on extrapulmonary epithelial surfaces and in serum, where it has been used as a biomarker for pulmonary disease states. In this study, we investigate the mechanisms ...

  7. Protein deficiency in pregnant rats causes decreased levels of plasma somatomedin and its carrier protein associated with reduced plasma levels of placental lactogen and hepatic lactogenic receptor number.

    Science.gov (United States)

    Pilistine, S J; Munro, H N

    1984-03-01

    Rats were fed either a 20% lactalbumin (control) or a 5% lactalbumin (low protein) diet for the last 2 weeks of pregnancy. At day 20 of gestation, rat serum placental lactogen levels, measured by radioreceptor assay, were significantly decreased by the low protein diet, thus confirming our earlier findings. The number of microsomal membrane lactogenic receptors, measured on the maternal livers at the end of pregnancy, was severely reduced in the livers of the low protein group, whereas protein deficiency did not affect binding affinity. Serum concentrations of somatomedin, measured by a competitive binding assay after acid treatment of the serum to remove endogenous carrier protein, were extensively reduced in the low protein group. The amounts of the somatomedin carrier proteins in the serum were assayed by separation on Sephacryl-S300 columns into higher- and lower-molecular-weight fractions peak 2 and peak 3, respectively. For the low protein diet group, both fractions showed a reduction in binding capacity, more marked in the case of peak 2. Since placental lactogen is known to influence output of somatomedin by the liver, we hypothesize that protein deficiency during pregnancy causes a fall in serum somatomedin level by reducing secretion of placental lactogen, which regulates its production by the liver.

  8. Multifactorial level of extremostability of proteins: can they be exploited for protein engineering?

    Science.gov (United States)

    Chakravorty, Debamitra; Khan, Mohd Faheem; Patra, Sanjukta

    2017-03-10

    Research on extremostable proteins has seen immense growth in the past decade owing to their industrial importance. Basic research of attributes related to extreme-stability requires further exploration. Modern mechanistic approaches to engineer such proteins in vitro will have more impact in industrial biotechnology economy. Developing a priori knowledge about the mechanism behind extreme-stability will nurture better understanding of pathways leading to protein molecular evolution and folding. This review is a vivid compilation about all classes of extremostable proteins and the attributes that lead to myriad of adaptations divulged after an extensive study of 6495 articles belonging to extremostable proteins. Along with detailing on the rationale behind extreme-stability of proteins, emphasis has been put on modern approaches that have been utilized to render proteins extremostable by protein engineering. It was understood that each protein shows different approaches to extreme-stability governed by minute differences in their biophysical properties and the milieu in which they exist. Any general rule has not yet been drawn regarding adaptive mechanisms in extreme environments. This review was further instrumental to understand the drawback of the available 14 stabilizing mutation prediction algorithms. Thus, this review lays the foundation to further explore the biophysical pleiotropy of extreme-stable proteins to deduce a global prediction model for predicting the effect of mutations on protein stability.

  9. Serum Protein Thiol Levels in Patients with Hospital-Acquired Acute Kidney Injury

    Directory of Open Access Journals (Sweden)

    Jing Qian

    2015-12-01

    Full Text Available Background/Aims: This study aimed to examine antioxidants in patients with acute kidney injury (AKI and determine whether serum protein thiol levels are associated with all-cause 90-day mortality in patients with hospital-acquired AKI. Methods: According to the RIFLE criteria, 160 patients with hospital-acquired AKI were enrolled in our prospective cohort study. As controls, 72 critically ill patients without AKI and 72 age and sex-matched healthy subjects were also recruited. Serum protein thiol levels were analyzed in relation to all-cause mortality of patients with AKI. Results: Serum protein thiol levels in AKI patients were lower than those in healthy people (p=0.010. Protein thiol levels showed a weak but significant positive correlation with serum albumin levels. The 90-day overall mortality rate was higher in AKI patients with high serum protein thiol levels than in those with low levels (p=0.032 by log rank test. In multivariate analysis (Cox regression, serum protein thiol levels (p=0.031 were independently associated with 90-day overall mortality after adjustment for age, sex, sepsis, and the Acute Physiology and Chronic Health Evaluation II score. Conclusions: Patients with hospital-acquired AKI have remarkably low serum protein thiol levels. Elevated protein thiol levels are associated with 90-day overall mortality in hospital-acquired AKI.

  10. Desensitization of G-protein-coupled receptors induces vascular hypocontractility in response to norepinephrine in the mesenteric arteries of cirrhotic patients and rats

    Institute of Scientific and Technical Information of China (English)

    Wei Chen; Jiang-Yong Sang; De-Jun Liu; Jun Qin; Yan-Miao Huo; Jia Xu and Zhi-Yong Wu

    2013-01-01

    BACKGROUND: The  increased  β-arrestin-2  and  its  combina-tion with G-protein-coupled receptors (GPCRs) lead to GPCRs desensitization.  The  latter  may  be  responsible  for  decreased contractile  reactivity  in  the  mesenteric  arteries  of  cirrhotic patients  and  rats.  The  present  study  is  to  investigate  the machinery  changes  of  α-adrenergic  receptors  and  G  proteins and  their  roles  in  the  contractility  of  mesenteric  arteries  of cirrhotic patients and animal models. METHODS: Patients  with  cirrhosis  due  to  hepatitis  B  and cirrhotic rats induced by CCl4 were studied. Mesenteric artery contractility  in  response  to  norepinephrine  was  determined by  a  vessel  perfusion  system.  The  contractile  effect  of  G protein-coupled  receptor  kinase-2  (GRK-2)  inhibitor  on  the mesenteric artery was evaluated. The protein expression of the α1 adrenergic receptor, G proteins, β-arrestin-2, GRK-2 as well as  the  activity  of  Rho  associated  coiled-coil  forming  protein kinase-1 (ROCK-1) were measured by Western blot. In addition, the interaction of α1 adrenergic receptor with β-arrestin-2 was assessed by co-immunoprecipitation. RESULTS: The  portal  vein  pressure  of  cirrhotic  patients  and rats  was  significantly  higher  than  that  of  controls.  The  dose-response curve to norepinephrine in mesenteric arteriole was shifted  to  the  right,  and  EC50  was  significantly  increased  in cirrhotic patients and rats. There were no significant differences in the expressions of the α1 adrenergic receptor and G

  11. GUN1 Controls Accumulation of the Plastid Ribosomal Protein S1 at the Protein Level and Interacts with Proteins Involved in Plastid Protein Homeostasis.

    Science.gov (United States)

    Tadini, Luca; Pesaresi, Paolo; Kleine, Tatjana; Rossi, Fabio; Guljamow, Arthur; Sommer, Frederik; Mühlhaus, Timo; Schroda, Michael; Masiero, Simona; Pribil, Mathias; Rothbart, Maxi; Hedtke, Boris; Grimm, Bernhard; Leister, Dario

    2016-03-01

    Developmental or metabolic changes in chloroplasts can have profound effects on the rest of the plant cell. Such intracellular responses are associated with signals that originate in chloroplasts and convey information on their physiological status to the nucleus, which leads to large-scale changes in gene expression (retrograde signaling). A screen designed to identify components of retrograde signaling resulted in the discovery of the so-called genomes uncoupled (gun) mutants. Genetic evidence suggests that the chloroplast protein GUN1 integrates signals derived from perturbations in plastid redox state, plastid gene expression, and tetrapyrrole biosynthesis (TPB) in Arabidopsis (Arabidopsis thaliana) seedlings, exerting biogenic control of chloroplast functions. However, the molecular mechanism by which GUN1 integrates retrograde signaling in the chloroplast is unclear. Here we show that GUN1 also operates in adult plants, contributing to operational control of chloroplasts. The gun1 mutation genetically interacts with mutations of genes for the chloroplast ribosomal proteins S1 (PRPS1) and L11. Analysis of gun1 prps1 lines indicates that GUN1 controls PRPS1 accumulation at the protein level. The GUN1 protein physically interacts with proteins involved in chloroplast protein homeostasis based on coimmunoprecipitation experiments. Furthermore, yeast two-hybrid and bimolecular fluorescence complementation experiments suggest that GUN1 might transiently interact with several TPB enzymes, including Mg-chelatase subunit D (CHLD) and two other TPB enzymes known to activate retrograde signaling. Moreover, the association of PRPS1 and CHLD with protein complexes is modulated by GUN1. These findings allow us to speculate that retrograde signaling might involve GUN1-dependent formation of protein complexes.

  12. Real-time quantification of protein expression at the single-cell level via dynamic protein synthesis translocation reporters.

    Science.gov (United States)

    Aymoz, Delphine; Wosika, Victoria; Durandau, Eric; Pelet, Serge

    2016-04-21

    Protein expression is a dynamic process, which can be rapidly induced by extracellular signals. It is widely appreciated that single cells can display large variations in the level of gene induction. However, the variability in the dynamics of this process in individual cells is difficult to quantify using standard fluorescent protein (FP) expression assays, due to the slow maturation of their fluorophore. Here we have developed expression reporters that accurately measure both the levels and dynamics of protein synthesis in live single cells with a temporal resolution under a minute. Our system relies on the quantification of the translocation of a constitutively expressed FP into the nucleus. As a proof of concept, we used these reporters to measure the transient protein synthesis arising from two promoters responding to the yeast hyper osmolarity glycerol mitogen-activated protein kinase pathway (pSTL1 and pGPD1). They display distinct expression dynamics giving rise to strikingly different instantaneous expression noise.

  13. The effect of serum magnesium levels and serum endothelin-1 levels on bone mineral density in protein energy malnutrition.

    Science.gov (United States)

    Ozturk, C F; Karakelleoglu, C; Orbak, Z; Yildiz, L

    2012-06-01

    An inadequate and imbalanced intake of protein and energy results in protein-energy malnutrition (PEM). It is known that bone mineral density and serum magnesium levels are low in malnourished children. However, the roles of serum magnesium and endothelin-1 (ET-1) levels in the pathophysiology of bone mineralization are obscure. Thus, the relationships between serum magnesium and ET-1 levels and the changes in bone mineral density were investigated in this study. There was a total of 32 subjects, 25 of them had PEM and seven were controls. While mean serum ET-1 levels of the children with kwashiorkor and marasmus showed no statistically significant difference, mean serum ET-1 levels of both groups were significantly higher than that of the control group. Serum magnesium levels were lower than normal value in 9 (36%) of 25 malnourished children. Malnourished children included in this study were divided into two subgroups according to their serum magnesium levels. While mean serum ET-1 levels in the group with low magnesium levels were significantly higher than that of the group with normal magnesium levels (p malnutrition. Our study suggested that lower magnesium levels and higher ET-1 levels might be important factors in changes of bone mineral density in malnutrition. We recommend that the malnourished patients, especially with hypomagnesaemia, should be treated with magnesium early.

  14. Effect of nonsteroidal antiinflammatory drugs on the C-reactive protein level in rheumatoid arthritis

    DEFF Research Database (Denmark)

    Tarp, Simon; Bartels, Else M; Bliddal, Henning;

    2012-01-01

    To evaluate the effects of oral nonsteroidal antiinflammatory drugs (NSAIDs) on C-reactive protein (CRP) levels in rheumatoid arthritis (RA) patients, with a prespecified focus on the different NSAIDs.......To evaluate the effects of oral nonsteroidal antiinflammatory drugs (NSAIDs) on C-reactive protein (CRP) levels in rheumatoid arthritis (RA) patients, with a prespecified focus on the different NSAIDs....

  15. Effects of dietary protein level on growth, health and physiological parameters in growing-furring mink

    DEFF Research Database (Denmark)

    Damgaard, Birthe Marie; Larsen, Peter F.; Clausen, Tove

    2012-01-01

    The aim of the study was to investigate the effects of the dietary protein level and the feeding strategy on growth, health and physiological blood and liver parameters in growing-furring male mink. Effects of dietary protein levels ranging from 22% of metabolizable energy (MEp) to experimental p...

  16. Development, Characterization, and Optimization of Protein Level in Date Bars Using Response Surface Methodology

    OpenAIRE

    Muhammad Nadeem; Salim-ur-Rehman,; Faqir Muhammad Anjum; Mian Anjum Murtaza; Ghulam Mueen-ud-Din

    2012-01-01

    This project was designed to produce a nourishing date bar with commercial value especially for school going children to meet their body development requirements. Protein level of date bars was optimized using response surface methodology (RSM). Economical and underutilized sources, that is, whey protein concentrate and vetch protein isolates, were explored for protein supplementation. Fourteen date bar treatments were produced using a central composite design (CCD) with 2 variables and 3 lev...

  17. High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells

    Institute of Scientific and Technical Information of China (English)

    Fei Zhong; Zhen-Yu Zhong; Shuang Liang; Xiu-Jin Li

    2006-01-01

    AIM: To develop a highly efficacious method for preparation of soluble SARS S-protein using adenovirus vector to meet the requirement for S-protein investigation.METHODS: The human adenovirus vector was used to express the soluble S-protein (corresponding to 1~1190 amino acids) fused with Myc/His tag using codon-optimized gene construct in HEK239 cells. The recombinant adenovirus bearing S-protein gene was generated by ligation method. The expressed S-protein with Myc/His tag was purified from culture medium with Ni-NTA agarose beads followed by dialysis. The S-protein was detected by Western blot and its biologic activity was analyzed by binding to Vero cells.RESULTS: Under the conditions of infection dose (MOI of 50) and expression time (48 h), the high-level expression of S-protein was obtained. The expression level was determined to be approximately 75 μg/106cells after purification. Purified soluble S-protein was readily detected by Western blot with anti-Myc antibody and showed the ability to bind to surface of Vero cells,demonstrating that the soluble S-protein could remain the biologic activity in the native molecule.CONCLUSION: The high-level expression of S-protein in HEK293 cells mediated by adenovirus can be achieved under the optimized expression conditions. The proteins possess the biologic activity, which lays a foundation for further investigation of S-protein biological function.

  18. Predicting protein folding pathways at the mesoscopic level based on native interactions between secondary structure elements

    Directory of Open Access Journals (Sweden)

    Sze Sing-Hoi

    2008-07-01

    Full Text Available Abstract Background Since experimental determination of protein folding pathways remains difficult, computational techniques are often used to simulate protein folding. Most current techniques to predict protein folding pathways are computationally intensive and are suitable only for small proteins. Results By assuming that the native structure of a protein is known and representing each intermediate conformation as a collection of fully folded structures in which each of them contains a set of interacting secondary structure elements, we show that it is possible to significantly reduce the conformation space while still being able to predict the most energetically favorable folding pathway of large proteins with hundreds of residues at the mesoscopic level, including the pig muscle phosphoglycerate kinase with 416 residues. The model is detailed enough to distinguish between different folding pathways of structurally very similar proteins, including the streptococcal protein G and the peptostreptococcal protein L. The model is also able to recognize the differences between the folding pathways of protein G and its two structurally similar variants NuG1 and NuG2, which are even harder to distinguish. We show that this strategy can produce accurate predictions on many other proteins with experimentally determined intermediate folding states. Conclusion Our technique is efficient enough to predict folding pathways for both large and small proteins at the mesoscopic level. Such a strategy is often the only feasible choice for large proteins. A software program implementing this strategy (SSFold is available at http://faculty.cs.tamu.edu/shsze/ssfold.

  19. Interactive Effects of Indigestible Carbohydrates, Protein Type, and Protein Level on Biomarkers of Large Intestine Health in Rats.

    Directory of Open Access Journals (Sweden)

    Marcin Taciak

    Full Text Available The effects of indigestible carbohydrates, protein type, and protein level on large intestine health were examined in rats. For 21 days, 12 groups of six 12-week-old male Wistar rats were fed diets with casein (CAS, or potato protein concentrate (PPC, providing 14% (lower protein level; LP, or 20% (higher protein level; HP protein, and containing cellulose, resistant potato starch, or pectin. Fermentation end-products, pH, and β-glucuronidase levels in cecal digesta, and ammonia levels in colonic digesta were determined. Cecal digesta, tissue weights, cecal and colon morphology, and colonocyte DNA damage were also analyzed. Digesta pH was lower, whereas relative mass of cecal tissue and digesta were higher in rats fed pectin diets than in those fed cellulose. Cecal parameters were greater in rats fed PPC and HP diets than in those fed CAS and LP diets, respectively. Short-chain fatty acid (SCFA concentrations were unaffected by protein or carbohydrate type. Total SCFA, acetic acid, and propionic acid concentrations were greater in rats fed LP diets than in those fed HP. Cecal pool of isobutyric and isovaleric acids was greater in rats fed PPC than in those fed CAS diets. PPC diets decreased phenol concentration and increased ammonia concentration in cecal and colonic digesta, respectively. Cecal crypt depth was greater in rats fed PPC and HP diets, and was unaffected by carbohydrates; whereas colonic crypt depth was greater in rats fed cellulose. Myenteron thickness in the cecum was unaffected by nutrition, but was greater in the colon of rats fed cellulose. Colonocyte DNA damage was greater in rats fed LP diets than in those fed HP diets, and was unaffected by carbohydrate or protein type. It was found that nutritional factors decreasing cecal digesta weight contribute to greater phenol production, increased DNA damage, and reduced ammonia concentration in the colon.

  20. Respon Pertumbuhan Ayam Lokal Pedaging terhadap Suplementasi Protein Isolasi Biji-bijian (PIB dan Perbedaan Level Protein Ransum

    Directory of Open Access Journals (Sweden)

    M. Aman Yaman

    2009-10-01

    Full Text Available The response of local meat chicken growth to supplementation of isolated grain protein and the difference in ration protein level ABSTRACT. A research which aims to determine the response of local meat chicken growth of protein supplementation with Isolation Grains Protein (IGB and the difference in ration protein level has been conducted in the Laboratory of Experimental Farm, Animal Husbandry Department, Faculty of Agriculture, Syiah Kuala University-Darussalam, Banda Aceh for 90 days. This study used a completely randomized design factorial with 2 factors, consisting of factors namely male gender (JJ and female (JB and the ration is a combination of factors and levels IGB in the ration, ie: treatment A: 17% protein and 0.4% IGB; treatment B 19% protein and 0.6% IGB and treatment C 21% protein and 0.8% IGB. Each combination consisted of 4 replications and each replication consists of 5 chickens. Parameters observed in the study were weight gain, achievement of final weight, consumption, conversion and efficiency of ration. DOC used a derivative result of selection of local meat chicken which are in the process of selection. Data acquired and processed by ANOVA. The results showed that supplementation of IGB and ration protein level difference was significantly effect (P <0.01 on weight gain, final weight, rate of consumption, conversion efficiency of rations and rations, but there is no interaction effect between sex and ration factors . The highest weight gain obtained in the male local chicken achieved by feeding a ration B (93.23 grams, while the hen rations achieved by providing treatment C (63.86 grams / week. The highest final body weight of male chicken on treatment B (1491.5 gram/90 days and hens in treatment C (1061.5 gram/90 days. However, the highest ration consumption in both male and female local chickens obtained from the ration A. Feed conversion value and the best feed efficiency obtained in treatment B for the treatment of

  1. Modulating protein interaction on a molecular and Microstructural level for texture control in protein based gels

    NARCIS (Netherlands)

    Martin, A.H.; Baigts Allende, D.; Munialo, C.D.; Urbonaite, V.; Pouvreau, L.A.M.; Jongh, de H.H.J.

    2014-01-01

    The exploration of microstructures and textures of protein based systems is essential to understand (oral) breakdown properties and thereby textural aspects, or macroscopic functionalities such as water holding. Upon structure breakdown, the applied energy (W) is primarily directed towards fracture

  2. Multi-level learning: improving the prediction of protein, domain and residue interactions by allowing information flow between levels

    Directory of Open Access Journals (Sweden)

    McDermott Drew

    2009-08-01

    Full Text Available Abstract Background Proteins interact through specific binding interfaces that contain many residues in domains. Protein interactions thus occur on three different levels of a concept hierarchy: whole-proteins, domains, and residues. Each level offers a distinct and complementary set of features for computationally predicting interactions, including functional genomic features of whole proteins, evolutionary features of domain families and physical-chemical features of individual residues. The predictions at each level could benefit from using the features at all three levels. However, it is not trivial as the features are provided at different granularity. Results To link up the predictions at the three levels, we propose a multi-level machine-learning framework that allows for explicit information flow between the levels. We demonstrate, using representative yeast interaction networks, that our algorithm is able to utilize complementary feature sets to make more accurate predictions at the three levels than when the three problems are approached independently. To facilitate application of our multi-level learning framework, we discuss three key aspects of multi-level learning and the corresponding design choices that we have made in the implementation of a concrete learning algorithm. 1 Architecture of information flow: we show the greater flexibility of bidirectional flow over independent levels and unidirectional flow; 2 Coupling mechanism of the different levels: We show how this can be accomplished via augmenting the training sets at each level, and discuss the prevention of error propagation between different levels by means of soft coupling; 3 Sparseness of data: We show that the multi-level framework compounds data sparsity issues, and discuss how this can be dealt with by building local models in information-rich parts of the data. Our proof-of-concept learning algorithm demonstrates the advantage of combining levels, and opens up

  3. Differential protein levels and post-translational modifications in spinal cord injury of the rat.

    Science.gov (United States)

    Afjehi-Sadat, Leila; Brejnikow, Mika; Kang, Sung Ung; Vishwanath, Vinay; Walder, Nadja; Herkner, Kurt; Redl, Heinz; Lubec, Gert

    2010-03-05

    Although changes in protein expression in spinal cord injury (SCI) would be of pivotal interest, information so far is limited. It was therefore the aim of the study to determine protein levels and post-translational modifications in the early phase following SCI in the rat. SCI was induced in Sprague-Dawley rats and sham operated rats served as controls. A gel-based proteomic approach using two-dimensional gel electrophoresis followed by quantification with specific software and subsequent identification of differentially expressed proteins by nano-ESI-LC-MS/MS was applied. Proteins of several pathways and cascades were dysregulated in SCI: 14-3-3 epsilon protein, dynein light chain 1, and tubulin beta-5 chain showed higher levels in SCI, whereas adenylyl cyclase associated protein 1, dihydropyrimidinase-related protein 2, F-actin capping protein subunit beta, glyceraldehyde-3-phosphate dehydrogenase, stress-induced phosphoprotein 1 and transthyretin showed lower levels in the injured tissue. Post-translational modifications indicated free oxygen radical attack on proteins in SCI. The occurrence of stress is indicated by deranged stress-induced phosphoprotein 1 and signaling abnormalities are reflected by adenylyl cyclase-associated protein 1 and 14-3-3 epsilon protein. The findings propose the involvement of the corresponding cascades and challenge further work into aberrant signaling and oxidative stress in SCI, which may form the basis for experimental intervention for spinal cord trauma.

  4. Genetic and environmental influences of surfactant protein D serum levels

    DEFF Research Database (Denmark)

    Sørensen, Grith Lykke; Hjelmborg, Jacob v. B.; Kyvik, Kirsten Ohm

    2006-01-01

    defining the constitutional serum level of SP-D and determine the magnitude of the genetic contribution to serum SP-D in the adult population. Recent studies have demonstrated that serum SP-D concentrations in children are genetically determined and that a single nucleotide polymorphism (SNP) located...... correlation was significantly higher for monozygotic (MZ) twin pairs than for dizygotic (DZ) twin pairs. Serum SP-D variance was influenced by nonshared environmental effects and additive genetic effects. Multivariate analysis of MZ and DZ covariance matrixes showed significant genetic correlation among serum...... SP-D and metabolic variables. The Met11Thr variant explained a significant part of the heritability indicating that serum SP-D variance could be decomposed into non-shared environmental effects (e(2) = 0.19), additive genetic effects (h(2) = 0.42), and the effect of the Met11Thr variations (q(2) = 0.39)....

  5. Gene-specific correlation of RNA and protein levels in human cells and tissues.

    Science.gov (United States)

    Edfors, Fredrik; Danielsson, Frida; Hallström, Björn M; Käll, Lukas; Lundberg, Emma; Pontén, Fredrik; Forsström, Björn; Uhlén, Mathias

    2016-10-20

    An important issue for molecular biology is to establish whether transcript levels of a given gene can be used as proxies for the corresponding protein levels. Here, we have developed a targeted proteomics approach for a set of human non-secreted proteins based on parallel reaction monitoring to measure, at steady-state conditions, absolute protein copy numbers across human tissues and cell lines and compared these levels with the corresponding mRNA levels using transcriptomics. The study shows that the transcript and protein levels do not correlate well unless a gene-specific RNA-to-protein (RTP) conversion factor independent of the tissue type is introduced, thus significantly enhancing the predictability of protein copy numbers from RNA levels. The results show that the RTP ratio varies significantly with a few hundred copies per mRNA molecule for some genes to several hundred thousands of protein copies per mRNA molecule for others. In conclusion, our data suggest that transcriptome analysis can be used as a tool to predict the protein copy numbers per cell, thus forming an attractive link between the field of genomics and proteomics.

  6. Influence of protein level and supplemental methionine in practical rations for young endangered masked bobwhite quail

    Science.gov (United States)

    Serafin, J.A.

    1982-01-01

    A study was conducted to examine the protein requirement of young endangered masked Bobwhite quail (Colinus virginianus ridgwayi). Five practical starting rations containing 24 to 32% protein were fed alone and supplemented with methionine for 5 weeks. Supplemental methionine significantly improved growth of quail fed diets containing 24 and 26% protein. Increasing the protein level improved growth of quail fed unsupplemented diets but did not do so when diets contained supplemental methionine. A methionine-supplemented ration containing 24% protein appeared adequate for supporting rapid growth of masked Bobwhite quail.

  7. Heart failure-specific changes in protein kinase signalling.

    Science.gov (United States)

    Lorenz, Kristina; Stathopoulou, Konstantina; Schmid, Evelyn; Eder, Petra; Cuello, Friederike

    2014-06-01

    Among the myriad of molecular alterations occurring in heart failure development, aggravation of the disease is often attributed to global or local changes in protein kinase activity, thus making protein kinases attractive targets for therapeutic intervention. Since protein kinases do not only have maladaptive roles, but also contribute to the physiological integrity of cells, it is a challenging task to circumvent undesired inhibition of protein kinase activity. Identification of posttranslational modifications and/or protein-protein interactions that are exclusively apparent under pathophysiological conditions provides exciting information for alternative non-kinase inhibitory treatment strategies that eliminate maladaptive functions of a protein kinase, but preserve the beneficial ones. Here, we focus on the disease-specific regulation of a number of protein kinases, namely, Ca(2+)/calmodulin-dependent protein kinase II isoform δ (CaMKIIδ), G protein-coupled receptor kinase 2 (GRK2), extracellular signal-regulated kinase 1 and 2 (ERK1/2), protein kinase D (PKD) and protein kinase C isoform β2 (PKCβ2), which are embedded in complex signal transduction pathways implicated in heart failure development, and discuss potential avenues for novel treatment strategies to combat heart disease.

  8. Development, Characterization, and Optimization of Protein Level in Date Bars Using Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Muhammad Nadeem

    2012-01-01

    Full Text Available This project was designed to produce a nourishing date bar with commercial value especially for school going children to meet their body development requirements. Protein level of date bars was optimized using response surface methodology (RSM. Economical and underutilized sources, that is, whey protein concentrate and vetch protein isolates, were explored for protein supplementation. Fourteen date bar treatments were produced using a central composite design (CCD with 2 variables and 3 levels for each variable. Date bars were then analyzed for nutritional profile. Proximate composition revealed that addition of whey protein concentrate and vetch protein isolates improved the nutritional profile of date bars. Protein level, texture, and taste were considerably improved by incorporating 6.05% whey protein concentrate and 4.35% vetch protein isolates in date bar without affecting any sensory characteristics during storage. Response surface methodology was observed as an economical and effective tool to optimize the ingredient level and to discriminate the interactive effects of independent variables.

  9. A novel method for increasing the expression level of recombinant proteins.

    Science.gov (United States)

    Wang, Aijun; Clapper, Jonathan; Guderian, Jeffery A; Foy, Teresa M; Fanger, Gary R; Retter, Marc W; Skeiky, Yasir A W

    2003-07-01

    Expression of recombinant proteins is an important step towards elucidating the functions of many genes discovered through genomic sequencing projects. It is also critical for validating gene targets and for developing effective therapies for many diseases. Here we describe a novel method to express recombinant proteins that are extremely difficult to produce otherwise. The increased protein expression level is achieved by using a fusion partner, MTB32-C, which is the carboxyl terminal fragment of the Mycobacterium tuberculosis antigen, MTB32 (Rv0125). By fusing MTB32-C to the N-termini of target genes, we have demonstrated significant enhancement of recombinant protein expression level in Escherichia coli. The inclusion of a 6xHis tag and the 128-amino acid of MTB32-C will add 13.5 kDa to the fusion molecule. Comparison of the mRNA levels of the fusion and non-fusion proteins indicated that the increased fusion protein expression may be regulated at translational or post-translational steps. There are many potential applications for the generated fusion proteins. For example, MTB32-C fusion proteins have been used successfully as immunogens to generate both polyclonal and monoclonal antibodies. These antibodies have been used to characterize cellular localization of the proteins and to validate gene targets at protein level. In addition, these antibodies may be useful in diagnostic and therapeutic applications for many diseases. If desired, the MTB32-C portion in the fusion protein can be removed after protein expression, making it possible to study protein structure and function as well as to screen for potential drugs. Thus, this novel fusion expression system has become a powerful tool for many applications.

  10. Synergistic Control of Kinetochore Protein Levels by Psh1 and Ubr2.

    Directory of Open Access Journals (Sweden)

    Eva Herrero

    2016-02-01

    Full Text Available The accurate segregation of chromosomes during cell division is achieved by attachment of chromosomes to the mitotic spindle via the kinetochore, a large multi-protein complex that assembles on centromeres. The budding yeast kinetochore comprises more than 60 different proteins. Although the structure and function of many of these proteins has been investigated, we have little understanding of the steady state regulation of kinetochores. The primary model of kinetochore homeostasis suggests that kinetochores assemble hierarchically from the centromeric DNA via the inclusion of a centromere-specific histone into chromatin. We tested this model by trying to perturb kinetochore protein levels by overexpressing an outer kinetochore gene, MTW1. This increase in protein failed to change protein recruitment, consistent with the hierarchical assembly model. However, we find that deletion of Psh1, a key ubiquitin ligase that is known to restrict inner kinetochore protein loading, does not increase levels of outer kinetochore proteins, thus breaking the normal kinetochore stoichiometry. This perturbation leads to chromosome segregation defects, which can be partially suppressed by mutation of Ubr2, a second ubiquitin ligase that normally restricts protein levels at the outer kinetochore. Together these data show that Psh1 and Ubr2 synergistically control the amount of proteins at the kinetochore.

  11. Association between protein C levels and mortality in patients with advanced prostate, lung and pancreatic cancer.

    Science.gov (United States)

    Wilts, I T; Hutten, B A; Meijers, J C M; Spek, C A; Büller, H R; Kamphuisen, P W

    2017-06-01

    Procoagulant factors promote cancer progression and metastasis. Protein C is involved in hemostasis, inflammation and signal transduction, and has a protective effect on the endothelial barrier. In mice, administration of activated protein C reduced experimental metastasis. We assessed the association between protein C and mortality in patients with three types of cancer. The study population consisted of patients with advanced prostate, non-small cell lung or pancreatic cancer, who participated in the INPACT trial (NCT00312013). The trial evaluated the addition of nadroparin to chemotherapy in patients with advanced malignancy. Patients were divided into tertiles based on protein C at baseline. The association between protein C levels and mortality was evaluated with Cox proportional hazard models. We analysed 477 patients (protein C tertiles: C level was 107% (IQR 92-129). In the lowest tertile, 75 patients per 100 patient-years died, as compared to 60 and 54 in the middle and high tertile, respectively. Lower levels of protein C were associated with increased mortality (in tertiles: HR for trend 1.18, 95%CI 1.02-1.36, adjusted for age, sex and nadroparin use; as a continuous variable: HR 1.004, 95%CI 1.00-1.008, p=0.07). Protein C seems inversely associated with mortality in patients with advanced prostate, lung and pancreatic cancer. Further research should validate protein C as a biomarker for mortality, and explore the effects of protein C on progression of cancer. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Synergistic Control of Kinetochore Protein Levels by Psh1 and Ubr2.

    Directory of Open Access Journals (Sweden)

    Eva Herrero

    2016-02-01

    Full Text Available The accurate segregation of chromosomes during cell division is achieved by attachment of chromosomes to the mitotic spindle via the kinetochore, a large multi-protein complex that assembles on centromeres. The budding yeast kinetochore comprises more than 60 different proteins. Although the structure and function of many of these proteins has been investigated, we have little understanding of the steady state regulation of kinetochores. The primary model of kinetochore homeostasis suggests that kinetochores assemble hierarchically from the centromeric DNA via the inclusion of a centromere-specific histone into chromatin. We tested this model by trying to perturb kinetochore protein levels by overexpressing an outer kinetochore gene, MTW1. This increase in protein failed to change protein recruitment, consistent with the hierarchical assembly model. However, we find that deletion of Psh1, a key ubiquitin ligase that is known to restrict inner kinetochore protein loading, does not increase levels of outer kinetochore proteins, thus breaking the normal kinetochore stoichiometry. This perturbation leads to chromosome segregation defects, which can be partially suppressed by mutation of Ubr2, a second ubiquitin ligase that normally restricts protein levels at the outer kinetochore. Together these data show that Psh1 and Ubr2 synergistically control the amount of proteins at the kinetochore.

  13. Effects of dietary digestible lysine levels on protein and fat deposition in the carcass of broilers

    Directory of Open Access Journals (Sweden)

    F de C Tavernari

    2009-06-01

    Full Text Available An experiment was carried out to evaluate the effects of different levels of digestible lysine in the diets of male and female broilers on protein and fat deposition. A total of 2160 Avian Farms broilers. A completely randomized experimental design was applied, and treatments consisted of the effects of three digestible lysine levels nested within each sex, with 12 replicates and 30 birds per experimental unit. The adopted digestible lysine levels corresponded to 92.5, 100.0, and 107.5% of the nutritional requirements of phases 1 to 21 days, 22 to 42 days, and 43 to 56 days of age, respectively. In each phase, the experimental diets contained similar calorie and protein levels within each sex. No significant effects of lysine levels were found on dry matter and fat percentages in the carcass of birds during the evaluated periods. Also, there were no significant effects of lysine levels on protein and fat deposition in males or females. However, males presented higher protein deposition and lower fat deposition than females during the total experimental period. Gompertz equations showed that females deposit more fat and less protein than males, and that this affected the fall in the curve of protein deposition, when the curve of fat deposition was still rising. Therefore, it was concluded that the older the broilers at slaughter, the higher their body fat content and the lower their body protein content, particularly in females.

  14. Protein levels of heme oxygenase-1 during reperfusion in human kidney transplants with delayed graft function.

    Science.gov (United States)

    Ollinger, Robert; Kogler, Pamela; Biebl, Matthias; Sieb, Michael; Sucher, Robert; Bösmüller, Claudia; Troppmair, Jakob; Mark, Walter; Weiss, Helmut; Margreiter, Raimund

    2008-01-01

    Delayed graft function (DGF) as a consequence of ischemia reperfusion injury (IRI) is associated with a decrease in long-term allograft survival. Heme oxygenase-1 (HO-1) is a stress responsive gene that is highly expressed in multiple pathological processes. The aim of our study was to analyze whether HO-1 protein levels in human kidney transplants during IRI correlate with the incidence of DGF. Kidney biopsies were obtained from 27 kidney allografts at two time points: at the end of cold storage and shortly after reperfusion. Samples were analyzed for HO-1 protein levels by Western blot. Heme oxygenase-1 protein levels were significantly higher in post-reperfusion biopsies (39.4 vs. 13.7 arbitrary units, p = 0.001). In pre-reperfusion biopsies no association was observed between HO-1 protein levels and DGF. In post-reperfusion biopsies, higher levels of HO-1 protein were measured in kidneys with DGF (53.7 vs. 36.2 arbitrary units, p = 0.064). DGF kidneys showed a significantly higher increase from pre- to post-reperfusion in HO-1 protein (42.0 vs. 18.7 arbitrary units, p = 0.025). Heme oxygenase-1 protein levels shortly after allograft reperfusion are closely related with initial graft function. Assessment thereof may be considered a valuable tool to predict DGF.

  15. Effects of Dietary Protein Level on Growth and Utilization of Protein and Energy by Juvenile Mangrove Red Snapper (Lutjanus argentimaculatus )

    Institute of Scientific and Technical Information of China (English)

    Ghulam Abbas; Khalid Jamil; Rukhsana Akhtar; Lin Hong

    2005-01-01

    A feeding trial was conducted in a recirculating water system to investigate the effects of dietary protein levels on growth, feed utilization, hepatosomatic index and liver lipid deposition of juvenile red snapper, Lutjanus argentimaculatus(average initial wet weight 8.0 ± 0.39 g and total length 3.14±0.3 cm). In the experiment, six fishmeal-based diets were formulated to contain various protein levels (20% to 45% in 5% increments), with dietary energy ranging from 2 210.7 kJ 100 g to 2 250.2 kJ 100 g dry matter. The protein to energy ratios of diets ranged from 8.58 mgprotein kJ-1 to 20.03 mgprotein kJ-1.Diets were fed for 90d to triplicate groups of fish stocked in 0.128 m3 seawater tanks, 25 individuals each. The daily ration of 2% wet body weight was offered to the fish thrice a day. The fish at the end of the study had more than ten-fold (77.0 g) increase in weight compared to the initial (8.0 g). Fish fed diets of 40% and 45% protein produced significantly (P < 0.05) higher weight gain of 77.2 g and 76.5 g, and specific growth rate (SGR) of 2.65% and 2.62% than those of 67.0 g and 68.3 g,and 2.49% and 2.51% of the other diets. The broken-line regression of SGR against dietary protein level yielded an optimum dietary protein requirement of 42.6% ( Y=- 1.6295 + 0.1114 X2, P < 0.05). Survival remained 100% among groups. Feed conversion ratio decreased from 0.45 for fish fed 20% dietary protein to 0.35 for fish fed 45% dietary protein. Nitrogen intake increased with an increase in dietary protein, which in turn resulted in an increase in nitrogen gain of fish whole body.Fish fed 40% and 45% protein diets showed higher (P < 0.05) nitrogen gain (0.27 g and 0.26 g) than those (0.23 g and 0.25 g)fed all other diets. Gross energy intake (GEI)in fish fed 45% protein was lower (600.67 kJ)than that (607.97 kJ) of 40%protein diet, though the differences were not statistically significant (P >0.05); GEI ranging from 677.31 kJ to 663.20 kJ at remaining four diets

  16. Epsilon PKC increases brain mitochondrial SIRT1 protein levels via heat shock protein 90 following ischemic preconditioning in rats.

    Directory of Open Access Journals (Sweden)

    John W Thompson

    Full Text Available Ischemic preconditioning is a neuroprotective mechanism whereby a sublethal ischemic exposure is protective against a subsequent lethal ischemic attack. We previously demonstrated that SIRT1, a nuclear localized stress-activated deacetylase, is vital for ischemic preconditioning neuroprotection. However, a recent study demonstrated that SIRT1 can also localize to the mitochondria. Mitochondrial localized SIRT1 may allow for a direct protection of mitochondria following ischemic preconditioning. The objective of this study was to determine whether ischemic preconditioning increases brain mitochondrial SIRT1 protein levels and to determine the role of PKCɛ and HSP90 in targeting SIRT1 to the mitochondria. Here we report that preconditioning rats, with 2 min of global cerebral ischemia, induces a delayed increase in non-synaptic mitochondrial SIRT1 protein levels which was not observed in synaptic mitochondria. This increase in mitochondrial SIRT1 protein was found to occur only in neuronal cells and was mediated by PKCε activation. Inhibition of HSP90, a protein chaperone involved in mitochondrial protein import, prevented preconditioning induced increases in mitochondrial SIRT1 and PKCε protein. Our work provides new insights into a possible direct role of SIRT1 in modulating mitochondrial function under both normal and stress conditions, and to a possible role of mitochondrial SIRT1 in activating preconditioning induced ischemic tolerance.

  17. Fasting Lipoprotein Lipase Protein Levels Can Predict a Postmeal Increment of Triglyceride Levels in Fasting Normohypertriglyceridemic Subjects.

    Science.gov (United States)

    Tsuzaki, Kokoro; Kotani, Kazuhiko; Yamada, Kazunori; Sakane, Naoki

    2016-09-01

    Although a postprandial increment in triglyceride (TG) levels is considered to be a risk factor for atherogenesis, tests (e.g., fat load) to assess postprandial changes in TG levels cannot be easily applied to clinical practice. Therefore, fasting markers that predict postprandial TG states are needed to be developed. One current candidate is lipoprotein lipase (LPL) protein, a molecule that hydrides TGs. This study investigated whether fasting LPL levels could predict postprandial TG levels. A total of 17 subjects (11 men, 6 women, mean age 52 ± 11 years) with normotriglyceridemia during fasting underwent the meal test. Several fasting parameters, including LPL, were measured for the area under the curve of postprandial TGs (AUC-TG). The subjects' mean fasting TG level was 1.30 mmol/l, and their mean LPL level was 41.6 ng/ml. The subjects' TG levels increased after loading (they peaked after two postprandial hours). Stepwise multiple regression analysis demonstrated that fasting TG levels were a predictor of the AUC-TG. In addition, fasting LPL mass levels were found to be a predictor of the AUC-TG (β = 0.65, P fasting TG levels. Fasting LPL levels may be useful to predict postprandial TG increment in this population. © 2015 Wiley Periodicals, Inc.

  18. The impact of carbohydrate and protein level and sources on swine manure foaming properties

    Science.gov (United States)

    This study explored the impact of swine diet on the composition, methane production potential, and foaming properties of manure. Samples of swine manure were collected from controlled feeding trials with diets varying in protein and carbohydrate levels and sources. Protein sources consisted of corn ...

  19. Effects of prebiotic, protein level, and stocking density on performance, immunity, and stress indicators of broilers

    National Research Council Canada - National Science Library

    Houshmand, M; Azhar, K; Zulkifli, I; Bejo, M H; Kamyab, A

    2012-01-01

    An experiment was conducted to determine the effects of period on the performance, immunity, and some stress indicators of broilers fed 2 levels of protein and stocked at a normal or high stocking density...

  20. the effect of dietary energy and protein levels on the composition of ...

    African Journals Online (AJOL)

    Zannel

    the diets of female ostriches on the composition of their eggs. .... Diet formulations were ..... although these trends were not evident in birds fed different protein levels. .... relation to nutrient-specific dietary supplements during egg laying.

  1. Modulation of GDP-fucose level for generating proteins with reduced rate of fucosylation (WO2010141855).

    Science.gov (United States)

    Taupin, Philippe

    2011-09-01

    The application (WO2010141855) is in the field of glycobiology, and involves the control of the rate of fucosylation of proteins by exogenous factors. It aims at controlling the rate of protein fucosylation with inhibitors (drugs or nucleic acid antagonists) of enzymes involved in the synthesis of GDP-fucose. Mammalian cell lines were cultured in the presence of inhibitors, for example, siRNA. The rates of GDP-fucose in cells and during protein fucosylation were characterized. The level of protein fucosylation decreases rapidly in response to a decrease in GDP-fucose level. The relationship between the rate of fucosylation of proteins and the level of GDP-fucose in a cell is non-linear. Reduction in the rate of protein fucosylation can be achieved with a minimal reduction of the level of GDP-fucose in cells. The paradigm may be used to synthesize proteins and antibodies, with a reduced rate of fucosylation. The application claims that the use of drugs or nucleic acid antagonists that inhibit the enzymes involved in GDP-fucose biosynthesis optimizes the level of GDP-fucose present in cells, and reduces the rate of fucosylation of glycoproteins.

  2. A Statistical Analysis of Protein-Protein Interaction with Knowledge-Based Potential at Residue Level

    Institute of Scientific and Technical Information of China (English)

    林巍; 孙飞; 饶子和

    2003-01-01

    Protein-protein recognition is an important step in biological processes, which still largely remains elusive.The inter-residue contact potential, CPij, describes the propensity of contact between two types of residue.In this study, several different CPij variants were examined with the objective of discriminating the binding potential of surface pairs.Using solvent mediated inter-molecule contact potential (SM-IMCPij), an evaluation model was deduced and tested.Using the evaluation model it was found that the SM-IMCPij gives a better performance than either residue mediated IMCPij(RM-IMCPij) or folding-residue contact potential (FCPij).The results suggest that the evaluation model provides a fast, effective, and discriminative method for the evaluation of proposed binding interfaces.

  3. Subunits of the Drosophila actin-capping protein heterodimer regulate each other at multiple levels.

    Directory of Open Access Journals (Sweden)

    Ana Rita Amândio

    Full Text Available The actin-Capping Protein heterodimer, composed of the α and β subunits, is a master F-actin regulator. In addition to its role in many cellular processes, Capping Protein acts as a main tumor suppressor module in Drosophila and in humans, in part, by restricting the activity of Yorkie/YAP/TAZ oncogenes. We aimed in this report to understand how both subunits regulate each other in vivo. We show that the levels and capping activities of both subunits must be tightly regulated to control F-actin levels and consequently growth of the Drosophila wing. Overexpressing capping protein α and β decreases both F-actin levels and tissue growth, while expressing forms of Capping Protein that have dominant negative effects on F-actin promote tissue growth. Both subunits regulate each other's protein levels. In addition, overexpressing one of the subunit in tissues knocked-down for the other increases the mRNA and protein levels of the subunit knocked-down and compensates for its loss. We propose that the ability of the α and β subunits to control each other's levels assures that a pool of functional heterodimer is produced in sufficient quantities to restrict the development of tumor but not in excess to sustain normal tissue growth.

  4. Subunits of the Drosophila actin-capping protein heterodimer regulate each other at multiple levels.

    Science.gov (United States)

    Amândio, Ana Rita; Gaspar, Pedro; Whited, Jessica L; Janody, Florence

    2014-01-01

    The actin-Capping Protein heterodimer, composed of the α and β subunits, is a master F-actin regulator. In addition to its role in many cellular processes, Capping Protein acts as a main tumor suppressor module in Drosophila and in humans, in part, by restricting the activity of Yorkie/YAP/TAZ oncogenes. We aimed in this report to understand how both subunits regulate each other in vivo. We show that the levels and capping activities of both subunits must be tightly regulated to control F-actin levels and consequently growth of the Drosophila wing. Overexpressing capping protein α and β decreases both F-actin levels and tissue growth, while expressing forms of Capping Protein that have dominant negative effects on F-actin promote tissue growth. Both subunits regulate each other's protein levels. In addition, overexpressing one of the subunit in tissues knocked-down for the other increases the mRNA and protein levels of the subunit knocked-down and compensates for its loss. We propose that the ability of the α and β subunits to control each other's levels assures that a pool of functional heterodimer is produced in sufficient quantities to restrict the development of tumor but not in excess to sustain normal tissue growth.

  5. Improved methodology for the affinity isolation of human protein complexes expressed at near endogenous levels

    DEFF Research Database (Denmark)

    Domanski, Michal; Molloy, Kelly; Jiang, Hua;

    2012-01-01

    An efficient and reliable procedure for the capture of affinity-tagged proteins and associated complexes from human cell lines is reported. Through multiple optimizations, high yield and low background affinity-purifications are achieved from modest quantities of human cells expressing endogenous......-level tagged proteins. Isolations of triple-FLAG and GFP-tagged fusion proteins involved in RNA metabolism are presented.......An efficient and reliable procedure for the capture of affinity-tagged proteins and associated complexes from human cell lines is reported. Through multiple optimizations, high yield and low background affinity-purifications are achieved from modest quantities of human cells expressing endogenous...

  6. Increased serum G72 protein levels in patients with schizophrenia: a potential candidate biomarker.

    Science.gov (United States)

    Akyol, Esra Soydaş; Albayrak, Yakup; Aksoy, Nurkan; Şahin, Başak; Beyazyüz, Murat; Kuloğlu, Murat; Hashimoto, Kenji

    2017-04-01

    The product of the G72 gene is an activator of d-amino acid oxidase and has been suggested to play a role in the pathogenesis of schizophrenia. Increased G72 protein levels may be associated with disturbed glutamatergic transmission and increased reactive oxygen species. Only one pilot study by Lin et al. has investigated the potential role of serum G72 protein levels as a biomarker for schizophrenia. In this study, we aimed to compare serum G72 protein levels between patients with schizophrenia and healthy controls, and to retest the results of the previous pilot study. Materials and methods In total, 107 patients with a diagnosis of schizophrenia according to the inclusion and exclusion criteria and 60 age-sex-matched healthy controls were included in the study. The groups were compared regarding serum G72 protein levels. The mean serum G72 protein values were 495.90±152.03 pg/ml in the schizophrenia group and 346.10±102.08 pg/ml in the healthy control group. The mean serum G72 protein level was significantly increased in the schizophrenia group compared with the healthy control group (t=-3.89, pschizophrenia and healthy control groups. It was determined that the cut-off value was 141.51 pg/ml with a sensitivity of 0.991 and a specificity of 0.821. We suggest that serum G72 protein levels may represent a candidate biomarker for schizophrenia and have confirmed the results of the previous preliminary study. Additional studies with larger sample sizes and the inclusion of first episode schizophrenia patients are required to clarify the reliability and validity of serum G72 protein levels as a biomarker for schizophrenia.

  7. Luciferase NanoLuc as a reporter for gene expression and protein levels in Saccharomyces cerevisiae

    Science.gov (United States)

    Masser, Anna E.; Kandasamy, Ganapathi; Kaimal, Jayasankar Mohanakrishnan

    2016-01-01

    Abstract Reporter proteins are essential tools in the study of biological processes and are employed to monitor changes in gene expression and protein levels. Luciferases are reporter proteins that enable rapid and highly sensitive detection with an outstanding dynamic range. Here we evaluated the usefulness of the 19 kDa luciferase NanoLuc (Nluc), derived from the deep sea shrimp Oplophorus gracilirostris, as a reporter protein in yeast. Cassettes with codon‐optimized genes expressing yeast Nluc (yNluc) or its destabilized derivative yNlucPEST have been assembled in the context of the dominant drug resistance marker kanMX. The reporter proteins do not impair the growth of yeast cells and exhibit half‐lives of 40 and 5 min, respectively. The commercial substrate Nano‐Glo® is compatible with detection of yNluc bioluminescence in bioluminescent signal and mRNA levels during both induction and decay. We demonstrated that the bioluminescence of yNluc fused to the C‐terminus of a temperature‐sensitive protein reports on its protein levels. In conclusion, yNluc and yNlucPEST are valuable new reporter proteins suitable for experiments with yeast using standard commercial substrate. © 2016 The Authors. Yeast published by John Wiley & Sons Ltd. PMID:26860732

  8. Effect of energy and protein levels on nutrient utilization and their requirements in growing Murrah buffaloes.

    Science.gov (United States)

    Prusty, Sonali; Kundu, Shivlal Singh; Mondal, Goutam; Sontakke, Umesh; Sharma, Vijay Kumar

    2016-04-01

    To evaluate different levels of energy and protein for optimum growth of Murrah male buffalo calves, a growth trial (150 days) was conducted on 30 calves (body weight 202.5 ± 6.8 kg). Six diets were formulated to provide 90, 100 and 110% protein level and 90 and 110% energy level requirements for buffalo calves, derived from ICAR 2013 recommendations for buffaloes. The crude protein (CP) intake was increased with higher dietary CP, whereas no effect of energy levels or interaction between protein and energy was observed on CP intake. There were significant effects (P interaction between protein and energy (P nutrient intake (protein or energy) per kg body weight (BW)(0.75) at various fortnight intervals was regressed linearly from the average daily gain (ADG) per kg BW(0.75). By setting the average daily gain at zero in the developed regression equation, a maintenance requirement was obtained, i.e. 133.1 kcal ME, 6.45 g CP and 3.95 g metabolizable protein (MP) per kg BW(0.75). Requirement for growth was 6.12 kcal ME, 0.46 g CP and 0.32 g MP per kg BW(0.75) per day. Metabolizable amino acid requirement was estimated from partitioning of MP intake and ADG. The ME requirements were lower, whereas the MP requirement of Murrah buffaloes was higher than ICAR (2013) recommendations.

  9. Multiple functions of G protein-coupled receptor kinases.

    Science.gov (United States)

    Watari, Kenji; Nakaya, Michio; Kurose, Hitoshi

    2014-03-06

    Desensitization is a physiological feedback mechanism that blocks detrimental effects of persistent stimulation. G protein-coupled receptor kinase 2 (GRK2) was originally identified as the kinase that mediates G protein-coupled receptor (GPCR) desensitization. Subsequent studies revealed that GRK is a family composed of seven isoforms (GRK1-GRK7). Each GRK shows a differential expression pattern. GRK1, GRK4, and GRK7 are expressed in limited tissues. In contrast, GRK2, GRK3, GRK5, and GRK6 are ubiquitously expressed throughout the body. The roles of GRKs in GPCR desensitization are well established. When GPCRs are activated by their agonists, GRKs phosphorylate serine/threonine residues in the intracellular loops and the carboxyl-termini of GPCRs. Phosphorylation promotes translocation of β-arrestins to the receptors and inhibits further G protein activation by interrupting receptor-G protein coupling. The binding of β-arrestins to the receptors also helps to promote receptor internalization by clathrin-coated pits. Thus, the GRK-catalyzed phosphorylation and subsequent binding of β-arrestin to GPCRs are believed to be the common mechanism of GPCR desensitization and internalization. Recent studies have revealed that GRKs are also involved in the β-arrestin-mediated signaling pathway. The GRK-mediated phosphorylation of the receptors plays opposite roles in conventional G protein- and β-arrestin-mediated signaling. The GRK-catalyzed phosphorylation of the receptors results in decreased G protein-mediated signaling, but it is necessary for β-arrestin-mediated signaling. Agonists that selectively activate GRK/β-arrestin-dependent signaling without affecting G protein signaling are known as β-arrestin-biased agonists. Biased agonists are expected to have potential therapeutic benefits for various diseases due to their selective activation of favorable physiological responses or avoidance of the side effects of drugs. Furthermore, GRKs are recognized as

  10. Proteomic analysis of day-night variations in protein levels in the rat pineal gland.

    Science.gov (United States)

    Møller, Morten; Sparre, Thomas; Bache, Nicolai; Roepstorff, Peter; Vorum, Henrik

    2007-06-01

    The pineal gland secretes the hormone melatonin. This secretion exhibits a circadian rhythm with a zenith during night and a nadir during day. We have performed proteome analysis of the superficial pineal gland in rats during daytime and nighttime. The proteins were extracted and subjected to 2-DE. Of 1747 protein spots revealed by electrophoresis, densitometric analysis showed the up-regulation of 25 proteins during nighttime and of 35 proteins during daytime. Thirty-seven of the proteins were identified by MALDI-TOF MS. The proteins up-regulated during the night are involved in the Krebs cycle, energy transduction, calcium binding, and intracellular transport. During the daytime, enzymes involved in glycolysis, electron transport, and also the Krebs cycle were up-regulated as well as proteins taking part in RNA binding and RNA processing. Our data show a prominent day-night variation of the protein levels in the rat pineal gland. Some proteins are up-regulated during the night concomitant with the melatonin secretion of the gland. Other proteins are up-regulated during the day indicating a pineal metabolism not related to the melatonin synthesis.

  11. Seroprofiling at the Candida albicans protein species level unveils an accurate molecular discriminator for candidemia.

    Science.gov (United States)

    Pitarch, Aida; Nombela, César; Gil, Concha

    2016-02-16

    Serum antibodies to specific Candida proteins have been reported as potential diagnostic biomarkers for candidemia. However, their diagnostic usefulness at the protein species level has hardly been examined. Using serological proteome analysis, we explored the IgG-antibody responses to Candida albicans protein species in candidemia and control patients. We found that 87 discrete protein species derived from 34 unique proteins were IgG-targets, although only 43 of them were differentially recognized by candidemia and control sera. An increase in the speciation of the immunome, connectivity and modularity of antigenic species co-recognition networks, and heterogeneity of antigenic species recognition patterns was associated with candidemia. IgG antibodies to certain discrete protein species were better predictors of candidemia than those to their corresponding proteins. A molecular discriminator delineated from the combined fingerprints of IgG antibodies to two distinct species of phosphoglycerate kinase and enolase accurately classified candidemia and control patients. These results provide new insight into the anti-Candida IgG-antibody response development in candidemia, and demonstrate that an immunoproteomic signature at the molecular level may be useful for its diagnosis. Our study further highlights the importance of defining pathogen-specific antigens at the chemical and molecular level for their potential application as immunodiagnostic reagents or even vaccine candidates.

  12. Performance of juvenile mojarra supplied with feed containing varying levels of crude protein

    Directory of Open Access Journals (Sweden)

    Ricardo Henrique Bastos de Souza

    2016-04-01

    Full Text Available ABSTRACT The growth of the Brazilian aquaculture has stimulated the development of the productive chain of native species, including marine environment. The objective of this study was to evaluate the growth performance of juvenile mojarra fish (Diapterus rhombeus fed diets containing different concentrations of crude protein (32, 36, 40 and 44 g 100 g-1. The 80 juvenile mojarra (7.2±1.5 g were kept in 16 circular tanks (150 L. The study design used was completely randomized with four treatments and four repetitions. The fish were fed four times a day. At the end of the experiment (60 days the final weight, feed intake, weight gain (WG, feed:gain ratio (FGR, protein efficiency rate (PER, energy efficiency rate, specific growth, survival rate and, body composition were evaluated. It was verified significant effect of protein level on the WG, with the best value at the level of 38.20 g 100 g-1 of crude protein. For FGR, the best estimated value occurred with 38.06 g 100 g-1 of crude protein, similar to that reported for the PER (38.91 g 100 g-1. The other performance parameters and body composition were not influenced by crude protein levels. Diet crude protein concentrations between 38.06 and 38.91 g 100 g-1 provide the best performance indices for juvenile mojarra.

  13. Zinc transporters protein level in postmortem brain of depressed subjects and suicide victims.

    Science.gov (United States)

    Rafalo-Ulinska, Anna; Piotrowska, Joanna; Kryczyk, Agata; Opoka, Włodzimierz; Sowa-Kucma, Magdalena; Misztak, Paulina; Rajkowska, Grazyna; Stockmeier, Craig A; Datka, Wojciech; Nowak, Gabriel; Szewczyk, Bernadeta

    2016-12-01

    Major depressive disorder (MDD) is a serious psychiatric illness, associated with an increasing rate of suicide. The pathogenesis of depression may be associated with the disruption of zinc (Zn) homeostasis. In the brain, several proteins that regulate Zn homeostasis are present, including Zn transporters (ZnTs) which remove Zn from the cytosol. The present study was designed to investigate whether depression and suicide are associated with alterations in the expression of the ZnTs protein. Protein levels of ZnT1, ZnT3, ZnT4, ZnT5 and ZnT6 were measured in postmortem brain tissue from two different cohorts. Cohort A contained 10 subjects diagnosed with MDD (7 were suicide victims) and 10 psychiatrically-normal control subjects and cohort B contained 11 non-diagnosed suicide victims and 8 sudden-death control subjects. Moreover, in cohort A we measured protein level of NMDA (GluN2A subunit), AMPA (GluA1 subunit) and 5-HT1A receptors and PSD-95. Proteins were measured in the prefrontal cortex (PFC) using Western blotting. In addition, Zn concentration was measured using a voltammetric method. There was a significant increase in protein levels of ZnT1, ZnT4, ZnT5 in the PFC in MDD, relative to control subjects, while ZnT3 protein level was decreased in MDD. There was no significant difference in the Zn concentration in the PFC between control and MDD subjects. Similarly, in the PFC of suicide victims (non-diagnosed), an increase in protein levels of ZnT1, ZnT4, ZnT5 and ZnT6 was observed. Conversely, protein levels of ZnT3 were decreased in both suicide victims and subjects with MDD, in comparison with control subjects. There was also a significant decrease in the protein level of GluA1, GluN2A, PSD-95 and 5-HT1A in MDD. Our studies suggest that alterations in Zn transport proteins are associated with the pathophysiology of MDD and suicide. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Intercellular Variability in Protein Levels from Stochastic Expression and Noisy Cell Cycle Processes.

    Science.gov (United States)

    Soltani, Mohammad; Vargas-Garcia, Cesar A; Antunes, Duarte; Singh, Abhyudai

    2016-08-01

    Inside individual cells, expression of genes is inherently stochastic and manifests as cell-to-cell variability or noise in protein copy numbers. Since proteins half-lives can be comparable to the cell-cycle length, randomness in cell-division times generates additional intercellular variability in protein levels. Moreover, as many mRNA/protein species are expressed at low-copy numbers, errors incurred in partitioning of molecules between two daughter cells are significant. We derive analytical formulas for the total noise in protein levels when the cell-cycle duration follows a general class of probability distributions. Using a novel hybrid approach the total noise is decomposed into components arising from i) stochastic expression; ii) partitioning errors at the time of cell division and iii) random cell-division events. These formulas reveal that random cell-division times not only generate additional extrinsic noise, but also critically affect the mean protein copy numbers and intrinsic noise components. Counter intuitively, in some parameter regimes, noise in protein levels can decrease as cell-division times become more stochastic. Computations are extended to consider genome duplication, where transcription rate is increased at a random point in the cell cycle. We systematically investigate how the timing of genome duplication influences different protein noise components. Intriguingly, results show that noise contribution from stochastic expression is minimized at an optimal genome-duplication time. Our theoretical results motivate new experimental methods for decomposing protein noise levels from synchronized and asynchronized single-cell expression data. Characterizing the contributions of individual noise mechanisms will lead to precise estimates of gene expression parameters and techniques for altering stochasticity to change phenotype of individual cells.

  15. [Serum Protein Expression Level in 47 Patients with Megaloblastic Anemia and Its Clinical Significance].

    Science.gov (United States)

    Ke, Yong

    2016-08-01

    To study the clinical significance of serum protein expression level in patients with megaloblastic anemia(MA). A total of 47 patients with MA were enrolled in this study between November 2013 and November 2015, and 50 healthy people in the same period were selected as controls. The levels of total protein (TP), albumin (Alb), ferritin (FER), transferrin (TRF) and soluble transferrin receptor (sTfR) were compared between 2 groups, and the serum protein expression levels in different types of MA, varous anemia degrees of MA were analyzed. The leves of TP, Alb and FER in MA patients were significantly lower than those in control group, the levels of TRF and sTfR were statistically significantly higher than those in control group(Panemia were significantly higher than those in the patients with moderate and severe anemia, the levels of TRF and sTfR were statistically significantly lower(Panemia were significantly higher than those in the patients with severe anemia, the levels of TRF and sTfR were significantly lower(P<0.05). Compared with levels before treatment, the levels of TP, Alb and FER significantly increased after treatment, while the TRF and sTfR levels significantly decreased (P<0.05). Serum levels of TP, Alb, FER, TRF and sTfR can provide a basis for the diagnosis of MA, and contribute to predict the disease to some extent.

  16. High levels of protein expression using different mammalian CMV promoters in several cell lines.

    Science.gov (United States)

    Xia, Wei; Bringmann, Peter; McClary, John; Jones, Patrick P; Manzana, Warren; Zhu, Ying; Wang, Soujuan; Liu, Yi; Harvey, Susan; Madlansacay, Mary Rose; McLean, Kirk; Rosser, Mary P; MacRobbie, Jean; Olsen, Catherine L; Cobb, Ronald R

    2006-01-01

    With the recent completion of the human genome sequencing project, scientists are faced with the daunting challenge of deciphering the function of these newly found genes quickly and efficiently. Equally as important is to produce milligram quantities of the therapeutically relevant gene products as quickly as possible. Mammalian expression systems provide many advantages to aid in this task. Mammalian cell lines have the capacity for proper post-translational modifications including proper protein folding and glycosylation. In response to the needs described above, we investigated the protein expression levels driven by the human CMV in the presence or absence of intron A, the mouse and rat CMV promoters with intron A, and the MPSV promoter in plasmid expression vectors. We evaluated the different promoters using an in-house plasmid vector backbone. The protein expression levels of four genes of interest driven by these promoters were evaluated in HEK293EBNA and CHO-K1 cells. Stable and transient transfected cells were utilized. In general, the full-length human CMV, in the presence of intron A, gave the highest levels of protein expression in transient transfections in both cell lines. However, the MPSV promoter resulted in the highest levels of stable protein expression in CHO-K1 cells. Using the CMV driven constitutive promoters in the presence of intron A, we have been able to generate >10 microg/ml of recombinant protein using transient transfections.

  17. A two level hierarchical model of protein retention in ion exchange chromatography.

    Science.gov (United States)

    Salvalaglio, Matteo; Paloni, Matteo; Guelat, Bertrand; Morbidelli, Massimo; Cavallotti, Carlo

    2015-09-11

    Predicting protein retention in ion exchange chromatography (IEX) from first principles is a fascinating perspective. In this work a two level hierarchical modeling strategy is proposed in order to calculate protein retention factors. Model predictions are tested against experimental data measured for Lysozyme and Chymotrypsinogen A in IEX columns as a function of ionic strength and pH. At the highest level of accuracy Molecular Dynamics (MD) simulations in explicit water are used to determine the interaction free energy between each of the two proteins and the IEX stationary phase for a reference pH and ionic strength. At a lower level of accuracy a linear response model based on an implicit treatment of solvation and adopting a static protein structure is used to calculate interaction free energies for the full range of pHs and ionic strengths considered. A scaling coefficient, determined comparing MD and implicit solvent simulations, is then introduced in order to correct the linear response model for errors induced by the adoption of a static protein structure. The calculated free energies are then used to compute protein retention factors, which can be directly compared with experimental data. The possibility to introduce a third level of accuracy is explored testing the predictions of a semiempirical model. A quantitative agreement between the predicted and measured protein retention factors is obtained using the coupled MD-linear response models, supporting the reliability of the proposed approach. The model allows quantifying the electrostatic, van der Waals, and conformational contributions to the interaction free energies. A good agreement between experiments and model is obtained also using the semiempirical model that, although requiring parameterization over higher level models or experimental data, proves to be useful in order to rapidly determine protein retention factors across wide pH and ionic strength ranges as it is computationally inexpensive.

  18. Acute Exercise Decreases Tribbles Homolog 3 Protein Levels in the Hypothalamus of Obese Rats.

    Science.gov (United States)

    Rodrigues, Barbara De Almeira; Pauli, Luciana Santos Souza; DE Souza, Claudio Teodoro; DA Silva, Adelino Sanchez Ramos; Cintra, Dennys Esper Correa; Marinho, Rodolfo; DE Moura, Leandro Pereira; Ropelle, Eloize Cristina Chiarreotto; Botezelli, José Diego; Ropelle, Eduardo Rochete; Pauli, José Rodrigo

    2015-08-01

    This study aims to evaluate the effects of acute exercise on tribbles homolog 3 (TRB3) protein levels and on the interaction between TRB3 and Akt proteins in the hypothalamus of obese rats. In addition, we evaluated the relationship between TRB3 and endoplasmic reticulum (ER) stress and verified whether an acute exercise session influences them. In the first part of the study, the rats were divided into three groups: control (lean), fed standard rodent chow; DIO, fed a high-fat diet; and DIO-EXE, fed a high-fat diet and submitted to a swimming acute exercise protocol. In the second part of the study, we used three other groups: control (lean) group receiving an intracerebroventricular (i.c.v.) infusion of vehicle, lean group receiving an i.c.v. infusion of thapsigargin, and lean group receiving an i.c.v. infusion of thapsigargin and performing an acute exercise session. Four hours after the exercise session, food intake was measured, and the hypothalamus was dissected and separated for subsequent protein analysis by immunoblotting and real-time polymerase chain reaction. The acute exercise session reduced TRB3 protein levels, disrupted the interaction between TRB3 and Akt proteins, increased the phosphorylation of Foxo1, and restored the anorexigenic effects of insulin on the hypothalamus of DIO rats. Interestingly, the suppressive effects of acute exercise on TRB3 protein levels may be related, at least in part, to decreased ER stress (evaluated though pancreatic ER kinase phosphorylation and C/EBP homologous protein levels) in the hypothalamus. Exercise-mediated reduction of hypothalamic TRB3 protein levels may be associated with reduction of ER stress. These data provide a new mechanism by which an acute exercise session improves insulin sensitivity in the hypothalamus and restores food intake control in obesity.

  19. The Effect of Dietary Crude Protein Level on Intestinal and Cecal Coccidiosis in Chicken

    Science.gov (United States)

    Sharma, V. D.; Fernando, M. A.; Summers, J. D.

    1973-01-01

    The effect of interaction of crude protein level in the diet and coccidiosis of the cecum and small intestine of chicks was investigated. A total of 390 day-old chicks were divided in 36 groups of ten and six groups of five chicks each. Twelve groups of ten and two groups of six chicks each were fed one of the three diets based on dietary crude protein level (16%, 20% and 24%). All diets contained an equal energy concentration. The chicks were on the appropriate diet for 15 days prior to infection. Each group was then subjected to one of the three treatments (a) control, (b) a single dose infection with 100,000 oocysts of Eimeria acervulina and (c) a single dose infection with 10,000 oocysts of Eimeria tenella. On the eighth day post infection all surviving E. tenella infected chicks and two replicates per dietary treatment of control and E. acervulina infected chicks were killed. An increase in dietary crude protein led to a linear (PCoccidiosis caused a reduction in daily gain, feed consumption and efficiency of feed utilization, the effect being more severe in E. tenella infection. The effect of dietary crude protein was protective against weight reduction. Chicks infected with E. tenella fed 24% crude protein had a higher (P<0.01) mortality rate than those fed on 16% or 20% crude protein level. The oocyst production by E. acervulina infected chicks was also higher (P<0.01) at the 24% crude protein level. The E. acervulina infected chicks exhibited compensatory growth during the eight to 14 days post infection. The compensatory growth was superior at the higher crude protein levels. The mechanism of compensatory growth is discussed. PMID:4266700

  20. Multi-level machine learning prediction of protein–protein interactions in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Julian Zubek

    2015-07-01

    Full Text Available Accurate identification of protein–protein interactions (PPI is the key step in understanding proteins’ biological functions, which are typically context-dependent. Many existing PPI predictors rely on aggregated features from protein sequences, however only a few methods exploit local information about specific residue contacts. In this work we present a two-stage machine learning approach for prediction of protein–protein interactions. We start with the carefully filtered data on protein complexes available for Saccharomyces cerevisiae in the Protein Data Bank (PDB database. First, we build linear descriptions of interacting and non-interacting sequence segment pairs based on their inter-residue distances. Secondly, we train machine learning classifiers to predict binary segment interactions for any two short sequence fragments. The final prediction of the protein–protein interaction is done using the 2D matrix representation of all-against-all possible interacting sequence segments of both analysed proteins. The level-I predictor achieves 0.88 AUC for micro-scale, i.e., residue-level prediction. The level-II predictor improves the results further by a more complex learning paradigm. We perform 30-fold macro-scale, i.e., protein-level cross-validation experiment. The level-II predictor using PSIPRED-predicted secondary structure reaches 0.70 precision, 0.68 recall, and 0.70 AUC, whereas other popular methods provide results below 0.6 threshold (recall, precision, AUC. Our results demonstrate that multi-scale sequence features aggregation procedure is able to improve the machine learning results by more than 10% as compared to other sequence representations. Prepared datasets and source code for our experimental pipeline are freely available for download from: http://zubekj.github.io/mlppi/ (open source Python implementation, OS independent.

  1. BDNF downregulates 5-HT(2A) receptor protein levels in hippocampal cultures

    DEFF Research Database (Denmark)

    Trajkovska, V; Santini, M A; Marcussen, Anders Bue;

    2009-01-01

    Both brain-derived neurotrophic factor (BDNF) and the serotonin receptor 2A (5-HT(2A)) have been related to depression pathology. Specific 5-HT(2A) receptor changes seen in BDNF conditional mutant mice suggest that BDNF regulates the 5-HT(2A) receptor level. Here we show a direct effect of BDNF...... on 5-HT(2A) receptor protein levels in primary hippocampal neuronal and mature hippocampal organotypic cultures exposed to different BDNF concentrations for either 1, 3, 5 or 7 days. In vivo effects of BDNF on hippocampal 5-HT(2A) receptor levels were further corroborated in (BDNF +/-) mice...... with reduced BDNF levels. In primary neuronal cultures, 7 days exposure to 25 and 50ng/mL BDNF resulted in downregulation of 5-HT(2A), but not of 5-HT(1A), receptor protein levels. The BDNF-associated downregulation of 5-HT(2A) receptor levels was also observed in mature hippocampal organotypic cultures...

  2. Effects of Different Protein Levels on the Growth Performance and Metabolic Rate of Nutrition in Broilers

    Institute of Scientific and Technical Information of China (English)

    WU Hongda

    2009-01-01

    The objective of this study is to determine the effect of different protein levels on the growth performance and metabolic rate of nutrition in broilers. Total 360 healthy and weight closed local broilers of 3 weeks were chosen and then divided into three groups randomly by one factor. Each group contains three handlings, each handling consists of 40 broilers. The period of experiment was 7 weeks. The metabolic experiment was performed at the 7th week. Three different protein levels were used in broilers' diet. The levels of protein were 19%, 17.5% and 16%. The results showed that the different levels of protein in crude dietary had significant difference between 19% group and the other two groups. The average daily weight gain and daily efficiency were significantly higher than that of the other two groups (P0.05), and the metabolic rate of the impact of phosphorus was significantly different (P<0.05). The result showed that when protein level was 19%, the growth of the local broiler was the best.

  3. A non-sequence-specific requirement for SMN protein activity: the role of aminoglycosides in inducing elevated SMN protein levels.

    Science.gov (United States)

    Wolstencroft, Elizabeth C; Mattis, Virginia; Bajer, Anna A; Young, Philip J; Lorson, Christian L

    2005-05-01

    Spinal muscular atrophy (SMA) is caused by homozygous loss of the survival motor neuron (SMN1) gene. In virtually all SMA patients, a nearly identical copy gene is present, SMN2. SMN2 cannot fully compensate for the loss of SMN1 because the majority of transcripts derived from SMN2 lack a critical exon (exon 7), resulting in a dysfunctional SMN protein. Therefore, the critical distinction between a functional and a dysfunctional SMN protein is the inclusion or the exclusion of the exon 7 encoded peptide. To determine the role of the 16 amino acids encoded by SMN exon 7, a panel of synthetic mutations were transiently expressed in SMA patient fibroblasts and HeLa cells. Consistent with previous reports, the protein encoded by SMN exons 1-6 was primarily restricted to the nucleus. However, a variety of heterologous sequences fused to the C-terminus of SMN exons 1-6 allowed mutant SMN proteins to properly distribute to the cytoplasm and to the nuclear gems. These data demonstrate that the SMN exon 7 sequence is not specifically required, rather this region functions as a non-specific 'tail' that facilitates proper localization. Therefore, a possible means to restore additional activity to the SMNDelta7 protein could be to induce a longer C-terminus by suppressing recognition of the native stop codon. To address this possibility, aminoglycosides were examined for their ability to restore detectable levels of SMN protein in SMA patient fibroblasts. Aminoglycosides can suppress the accurate identification of translation termination codons in eukaryotic cells. Consistent with this, treatment of SMA patient fibroblasts with tobramycin and amikacin resulted in a quantitative increase in SMN-positive gems and an overall increase in detectable SMN protein. Taken together, this work describes the role of the critical exon 7 region and identifies a possible alternative approach for therapeutic intervention.

  4. AFM visualization at a single-molecule level of denaturated states of proteins on graphite.

    Science.gov (United States)

    Barinov, Nikolay A; Prokhorov, Valery V; Dubrovin, Evgeniy V; Klinov, Dmitry V

    2016-10-01

    Different graphitic materials are either already used or believed to be advantageous in biomedical and biotechnological applications, e.g., as biomaterials or substrates for sensors. Most of these applications or associated important issues, such as biocompatibility, address the problem of adsorption of protein molecules and, in particular the conformational state of the adsorbed protein molecule on graphite. High-resolution AFM demonstrates highly oriented pyrolytic graphite (HOPG) induced denaturation of four proteins of blood plasma, such as ferritin, fibrinogen, human serum albumin (HSA) and immunoglobulin G (IgG), at a single molecule level. Protein denaturation is accompanied by the decrease of the heights of protein globules and spreading of the denatured protein fraction on the surface. In contrast, the modification of HOPG with the amphiphilic oligoglycine-hydrocarbon derivative monolayer preserves the native-like conformation and provides even more mild conditions for the protein adsorption than typically used mica. Protein unfolding on HOPG may have universal character for "soft" globular proteins. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Alcohol-Binding Sites in Distinct Brain Proteins: The Quest for Atomic Level Resolution

    Science.gov (United States)

    Howard, Rebecca J.; Slesinger, Paul A.; Davies, Daryl L.; Das, Joydip; Trudell, James R.; Harris, R. Adron

    2011-01-01

    Defining the sites of action of ethanol on brain proteins is a major prerequisite to understanding the molecular pharmacology of this drug. The main barrier to reaching an atomic-level understanding of alcohol action is the low potency of alcohols, ethanol in particular, which is a reflection of transient, low-affinity interactions with their targets. These mechanisms are difficult or impossible to study with traditional techniques such as radioligand binding or spectroscopy. However, there has been considerable recent progress in combining X-ray crystallography, structural modeling, and site-directed mutagenesis to define the sites and mechanisms of action of ethanol and related alcohols on key brain proteins. We review such insights for several diverse classes of proteins including inwardly rectifying potassium, transient receptor potential, and neurotransmit-ter-gated ion channels, as well as protein kinase C epsilon. Some common themes are beginning to emerge from these proteins, including hydrogen bonding of the hydroxyl group and van der Waals interactions of the methylene groups of ethanol with specific amino acid residues. The resulting binding energy is proposed to facilitate or stabilize low-energy state transitions in the bound proteins, allowing ethanol to act as a “molecular lubricant” for protein function. We discuss evidence for characteristic, discrete alcohol-binding sites on protein targets, as well as evidence that binding to some proteins is better characterized by an interaction region that can accommodate multiple molecules of ethanol. PMID:21676006

  6. Quality of buffalo milk as affected by dietary protein level and flaxseed supplementation.

    Science.gov (United States)

    Santillo, A; Caroprese, M; Marino, R; Sevi, A; Albenzio, M

    2016-10-01

    The aim of the present research was to evaluate the effects of protein level and flaxseed supplementation on the yield and quality of buffalo milk. In particular, the fatty acid profile of milk from buffalo cows subjected to different diets has been investigated. A 2×3 factorial design was tested with buffalo cows receiving 2 dietary crude protein (CP) and 3 flaxseed (FS) supplementation levels. Treatments were (1) low dietary CP level [12% of dry matter (DM)] and no flaxseed supplementation (LP); (2) low dietary CP level (12% of DM) and low flaxseed supplementation (500g/d) (LPFS500); (3) low dietary CP level (12% of DM) and moderate flaxseed supplementation (1,000g/d) (LPFS1000); (4) moderate dietary CP level (15% of DM) and no flaxseed supplementation (MP); (5) moderate dietary CP level (15% of DM) and low flaxseed supplementation (500g/d) (MPFS500); and (6) moderate dietary CP level (15% of DM) and moderate flaxseed supplementation (1,000g/d) (MPFS1000). Milk protein and casein were affected by flaxseed supplementation being higher in MP, intermediate in LP, and lower in flaxseed-supplemented diets. However, the results from the present study highlighted that low protein diets sustained milk yield, protein, and casein synthesis in milk when whole flaxseed was administered. Short-chain fatty acids, in particular C8:0 and C10:0, were the lowest in milk from buffalo cows fed the highest level of flaxseed supplementation. Medium-chain fatty acids were the lowest in FS1000, intermediate in FS500, and the highest in the HP and LP groups. Long-chain fatty acids were the highest in FS1000, intermediate in FS500 groups, and the lowest in milk from buffalo receiving no flaxseed supplementation. Protein level of the diet influenced the percentage of C18:0, which was higher in MP than LP groups. Total conjugated linoleic acid content evidenced the same trend of long-chain fatty acids, with an increase of about 7% in FL500 and of 22% in FL1000 than the control. Apart from

  7. Evolutionary tuning of protein expression levels of a positively autoregulated two-component system.

    Directory of Open Access Journals (Sweden)

    Rong Gao

    2013-10-01

    Full Text Available Cellular adaptation relies on the development of proper regulatory schemes for accurate control of gene expression levels in response to environmental cues. Over- or under-expression can lead to diminished cell fitness due to increased costs or insufficient benefits. Positive autoregulation is a common regulatory scheme that controls protein expression levels and gives rise to essential features in diverse signaling systems, yet its roles in cell fitness are less understood. It remains largely unknown how much protein expression is 'appropriate' for optimal cell fitness under specific extracellular conditions and how the dynamic environment shapes the regulatory scheme to reach appropriate expression levels. Here, we investigate the correlation of cell fitness and output response with protein expression levels of the E. coli PhoB/PhoR two-component system (TCS. In response to phosphate (Pi-depletion, the PhoB/PhoR system activates genes involved in phosphorus assimilation as well as genes encoding themselves, similarly to many other positively autoregulated TCSs. We developed a bacteria competition assay in continuous cultures and discovered that different Pi conditions have conflicting requirements of protein expression levels for optimal cell fitness. Pi-replete conditions favored cells with low levels of PhoB/PhoR while Pi-deplete conditions selected for cells with high levels of PhoB/PhoR. These two levels matched PhoB/PhoR concentrations achieved via positive autoregulation in wild-type cells under Pi-replete and -deplete conditions, respectively. The fitness optimum correlates with the wild-type expression level, above which the phosphorylation output saturates, thus further increase in expression presumably provides no additional benefits. Laboratory evolution experiments further indicate that cells with non-ideal protein levels can evolve toward the optimal levels with diverse mutational strategies. Our results suggest that the natural

  8. Occupational exposure levels of bioaerosol components are associated with serum levels of the acute phase protein Serum Amyloid A in greenhouse workers

    DEFF Research Database (Denmark)

    Madsen, Anne Mette; Thilsing, Trine; Bælum, Jesper;

    2016-01-01

    to elevated levels of bioaerosols. The objective of this study is to assess whether greenhouse workers personal exposure to bioaerosol components was associated with serum levels of the acute phase proteins Serum Amyloid A (SAA) and C-reactive protein (CRP). METHODS: SAA and CRP levels were determined...

  9. Plasma levels of osteocalcin and retinol binding protein-4 in patients with medullary thyroid carcinoma

    Directory of Open Access Journals (Sweden)

    Jabar Lotfi

    2014-04-01

    Conclusion: According to difference between plasma levels of osteocalcin and retinol binding protein-4 in patients suffered of medullary thyroid carcinoma comparison with normal subjects, it can be said that, probably medullary thyroid carcinoma has effect on bone and adipose tissue metabolism, so osteocalcin and retinol binding protein-4 hormones have potential to be used for confirmation of diagnosis or following treatment of medullary thyroid carcinoma.

  10. High-Level Production of Heterologous Protein by Engineered Yeasts Grown in Cottage Cheese Whey

    OpenAIRE

    Maullu, Carlo; Lampis, Giorgio; Desogus, Alessandra; Ingianni, Angela; Rossolini, Gian Maria; Pompei, Raffaello

    1999-01-01

    Cottage cheese whey is a cheese industry by-product still rich in proteins and lactose. Its recycling is seldom cost-effective. In this work we show that the lactose-utilizing yeast Kluyveromyces lactis, engineered for production of recombinant human lysozyme, can be grown in cottage cheese whey, resulting in high-level production of the heterologous protein (125 μg/ml).

  11. Anti-carbamylated Protein Antibody Levels Correlate with Anti-Sa (Citrullinated Vimentin) Antibody Levels in Rheumatoid Arthritis.

    Science.gov (United States)

    Challener, Gregory J; Jones, Jonathan D; Pelzek, Adam J; Hamilton, B JoNell; Boire, Gilles; de Brum-Fernandes, Artur José; Masetto, Ariel; Carrier, Nathalie; Ménard, Henri A; Silverman, Gregg J; Rigby, William F C

    2016-02-01

    The presence of anticitrullinated protein antibodies (ACPA) in rheumatoid arthritis (RA) indicates a breach in immune tolerance. Recent studies indicate that this breach extends to homocitrullination of lysines with the formation of anti-carbamylated protein (anti-CarP) antibodies. We analyzed the clinical and serologic relationships of anti-CarP in 2 RA cohorts. Circulating levels of immunoglobulin G anti-CarP antibodies were determined by ELISA in established (Dartmouth-Hitchcock Medical Center) and early (Sherbrooke University Hospital Center) cohorts and evaluated for anticyclic citrullinated peptide antibodies (anti-CCP), specific ACPA, and rheumatoid factor (RF) levels using the Student t test and correlation analysis. We identified elevated anti-CarP antibodies titers in 47.0% of seropositive patients (Dartmouth, n = 164), with relationships to anti-CCP (p elevated anti-CarP antibodies; titers correlated to anti-CCP (p = 0.01) but not IgM-RF (p = 0.09). A strong correlation with anti-Sa was observed: 47.9% anti-Sa+ patients were anti-CarP antibodies+ versus only 25.4% anti-Sa- in the Sherbrooke cohort (p = 0.0002), and 62.6% anti-Sa+ patients versus 26.9% anti-Sa- were anti-CarP antibodies+ in Dartmouth (p anti-CarP antibody positivity. We also describe a surprising and unexpected association of anti-CarP with anti-Sa antibodies that could not be explained by cross-reactivity. Further, considerable heterogeneity exists between anti-CarP reactivity and other citrullinated peptide reactivity, raising the question of how the pathogenesis of antibody responses for carbamylated proteins and citrullinated proteins may be linked in vivo.

  12. The significance of C-reactive protein levels in women with premature rupture of membranes.

    Science.gov (United States)

    Ismail, M A; Zinaman, M J; Lowensohn, R I; Moawad, A H

    1985-02-15

    In a prospective study of 100 patients with preterm premature rupture of membranes, clinical chorioamnionitis was present in 18 and histologic chorioamnionitis was present in 63. Patients who were managed conservatively for premature rupture of membranes were monitored by C-reactive protein determination, white blood cell and differential counts, maternal temperature, and fetal heart tone. C-reactive protein was measured nephelometrically (Immuno-chemistry Analyzer II, Beckman). Elevated C-reactive protein levels correlated well with both the pathologic and the clinical diagnosis of chorioamnionitis. Elevated C-reactive protein levels (at least 12 to 24 hours before delivery) were more sensitive than other standard laboratory or clinical tests in predicting chorioamnionitis both by clinical and pathologic criteria. When C-reactive protein values were normal, clinical chorioamnionitis was rarely found, whereas pathologically diagnosed chorioamnionitis was found half of the time. We conclude that although the C-reactive protein level is a very sensitive predictor of infectious morbidity in premature rupture of membranes, its specificity is not high.

  13. Bimodal gold nanoparticle therapeutics for manipulating exogenous and endogenous protein levels in mammalian cells.

    Science.gov (United States)

    Muroski, Megan E; Kogot, Joshua M; Strouse, Geoffrey F

    2012-12-05

    A new advance in cell transfection protocol using a bimodal nanoparticle agent to selectively manipulate protein expression levels within mammalian cells is demonstrated. The nanoparticle based transfection approach functions by controlled release of gene regulatory elements from a 6 nm AuNP (gold nanoparticle) surface. The endosomal release of the regulatory elements from the nanoparticle surface results in endogenous protein knockdown simultaneously with exogenous protein expression for the first 48 h. The use of fluorescent proteins as the endogenous and exogenous signals for protein expression enables the efficiency of codelivery of siRNA (small interfering RNA) for GFP (green fluorescent protein) knockdown and a dsRed-express linearized plasmid for induction to be optically analyzed in CRL-2794, a human kidney cell line expressing an unstable green fluorescent protein. Delivery of the bimodal nanoparticle in cationic liposomes results in 20% GFP knockdown within 24 h of delivery and continues exhibiting knockdown for up to 48 h for the bimodal agent. Simultaneous dsRed expression is observed to initiate within the same time frame with expression levels reaching 34% after 25 days although cells have divided approximately 20 times, implying daughter cell transfection has occurred. Fluorescence cell sorting results in a stable colony, as demonstrated by Western blot analysis. The simultaneous delivery of siRNA and linearized plasmid DNA on the surface of a single nanocrystal provides a unique method for definitive genetic control within a single cell and leads to a very efficient cell transfection protocol.

  14. CRUDE PROTEIN LEVELS IN THE DIETS OF TAMBAQUI, COLOSSOMA MACROPOMUM (CUVIER, 1818, FINGERLINGS

    Directory of Open Access Journals (Sweden)

    CHARLYAN DE SOUSA LIMA

    2016-01-01

    Full Text Available Tambaqui is intensively farmed because of its production characteristics; however, there is a lack of information lacks about the nutritional requirements of this species. The present study aimed to evaluate the effects of various crude protein (CP levels in the diets of tambaqui fingerlings. A total of 750 fingerlings with an initial weight of 0.35 ± 0.09 g were selected in a completely randomized design with six treatments (experimental feeds with 20%, 24%, 28%, 32%, 36%, and 40% CP, five replicates, and 25 fish per experimental unit. Performance, survival rate, carcass yield, body composition, protein deposition, body fat, and nitrogen retention efficiency were evaluated after 45 days. CP levels did not affect the following: feed conversion, survival rate, moisture content, and carcass yield. However, with increasing CP levels, protein efficiency ratio decreased. Weight gain, feed conversion, and specific growth rate improved until CP levels of 31.57%, 28.90%, and 31.12%, respectively, were achieved. Quadratic effects of elevated CP levels on body fat and body fat deposition were observed at minimum CP levels of 26.55% and 23.77%, respectively; and on body protein, body protein deposition, and nitrogen retention efficiency at maximum CP levels of 29.26%, 32.50%, and 27.21%, respectively. We conclude that a CP level of 31.57% is recommended for the diets of tambaqui fingerlings weighing between 0.35 and 15.11 g, which corresponds to a digestible energy:CP ratio of 9.50 kcal DE/g CP.

  15. Stochastic protein expression in individual cells at the single molecule level

    Science.gov (United States)

    Cai, Long; Friedman, Nir; Xie, X. Sunney

    2006-03-01

    In a living cell, gene expression-the transcription of DNA to messenger RNA followed by translation to protein-occurs stochastically, as a consequence of the low copy number of DNA and mRNA molecules involved. These stochastic events of protein production are difficult to observe directly with measurements on large ensembles of cells owing to lack of synchronization among cells. Measurements so far on single cells lack the sensitivity to resolve individual events of protein production. Here we demonstrate a microfluidic-based assay that allows real-time observation of the expression of β-galactosidase in living Escherichia coli cells with single molecule sensitivity. We observe that protein production occurs in bursts, with the number of molecules per burst following an exponential distribution. We show that the two key parameters of protein expression-the burst size and frequency-can be either determined directly from real-time monitoring of protein production or extracted from a measurement of the steady-state copy number distribution in a population of cells. Application of this assay to probe gene expression in individual budding yeast and mouse embryonic stem cells demonstrates its generality. Many important proteins are expressed at low levels, and are thus inaccessible by current genomic and proteomic techniques. This microfluidic single cell assay opens up possibilities for system-wide characterization of the expression of these low copy number proteins.

  16. Physiological enzymology: The next frontier in understanding protein structure and function at the cellular level.

    Science.gov (United States)

    Lee, Irene; Berdis, Anthony J

    2016-01-01

    Historically, the study of proteins has relied heavily on characterizing the activity of a single purified protein isolated from other cellular components. This classic approach allowed scientists to unambiguously define the intrinsic kinetic and chemical properties of that protein. The ultimate hope was to extrapolate this information toward understanding how the enzyme or receptor behaves within its native cellular context. These types of detailed in vitro analyses were necessary to reduce the innate complexities of measuring the singular activity and biochemical properties of a specific enzyme without interference from other enzymes and potential competing substrates. However, recent developments in fields encompassing cell biology, molecular imaging, and chemical biology now provide the unique chemical tools and instrumentation to study protein structure, function, and regulation in their native cellular environment. These advancements provide the foundation for a new field, coined physiological enzymology, which quantifies the function and regulation of enzymes and proteins at the cellular level. In this Special Edition, we explore the area of Physiological Enzymology and Protein Function through a series of review articles that focus on the tools and techniques used to measure the cellular activity of proteins inside living cells. This article is part of a Special Issue entitled: Physiological Enzymology and Protein Functions.

  17. Ground level environmental protein concentrations in various ecuadorian environments: potential uses of aerosolized protein for ecological research

    Science.gov (United States)

    Staton, Sarah J.R.; Woodward, Andrea; Castillo, Josemar A.; Swing, Kelly; Hayes, Mark A.

    2014-01-01

    Large quantities of free protein in the environment and other bioaerosols are ubiquitous throughout terrestrial ground level environments and may be integrative indicators of ecosystem status. Samples of ground level bioaerosols were collected from various ecosystems throughout Ecuador, including pristine humid tropical forest (pristine), highly altered secondary humid tropical forest (highly altered), secondary transitional very humid forest (regrowth transitional), and suburban dry montane deforested (suburban deforested). The results explored the sensitivity of localized aerosol protein concentrations to spatial and temporal variations within ecosystems, and their value for assessing environmental change. Ecosystem specific variations in environmental protein concentrations were observed: pristine 0.32 ± 0.09 μg/m3, highly altered 0.07 ± 0.05 μg/m3, regrowth transitional 0.17 ± 0.06 μg/m3, and suburban deforested 0.09 ± 0.04 μg/m3. Additionally, comparisons of intra-environmental differences in seasonal/daily weather (dry season 0.08 ± 0.03 μg/m3 and wet season 0.10 ± 0.04 μg/m3), environmental fragmentation (buffered 0.19 ± 0.06 μg/m3 and edge 0.15 ± 0.06 μg/m3), and sampling height (ground level 0.32 ± 0.09 μg/m3 and 10 m 0.24 ± 0.04 μg/m3) demonstrated the sensitivity of protein concentrations to environmental conditions. Local protein concentrations in altered environments correlated well with satellite-based spectral indices describing vegetation productivity: normalized difference vegetation index (NDVI) (r2 = 0.801), net primary production (NPP) (r2 = 0.827), leaf area index (LAI) (r2 = 0.410). Moreover, protein concentrations distinguished the pristine site, which was not differentiated in spectral indices, potentially due to spectral saturation typical of highly vegetated environments. Bioaerosol concentrations represent an inexpensive method to increase understanding of environmental changes, especially in densely vegetated

  18. mRNA and Protein levels of rat pancreas specific protein disulphide isomerase are downregulated during Hyperglycemia.

    Science.gov (United States)

    Gupta, Rajani; Bhar, Kaushik; Sen, Nandini; Bhowmick, Debajit; Mukhopadhyay, Satinath; Panda, Koustubh; Siddhanta, Anirban

    2016-02-01

    Diabetes (Type I and Type II) which affects nearly every organ in the body is a multi-factorial non-communicable disorder. Hyperglycemia is the most characteristic feature of this disease. Loss of beta cells is common in both types of diabetes whose detailed cellular and molecular mechanisms are yet to be elucidated. As this disease is complex, identification of specific biomarkers for its early detection, management and devising new therapies is challenging. Based on the fact that functionally defective proteins provide the biochemical basis for many diseases, in this study, we tried to identify differentially expressed proteins during hyperglycemia. For that, hyperglycemia was induced in overnight fasted rats by intra-peritoneal injection of streptozotocin (STZ). The pancreas was isolated from control and treated rats for subsequent analyses. The 2D-gel electrophoresis followed by MALDI-TOF-MS-MS analyses revealed several up- and down-regulated proteins in hyperglycemic rat pancreas including the downregulation of a pancreas specific isoform of protein disulphide isomerase a2 (Pdia2).This observation was validated by western blot. Quantitative PCR experiments showed that the level of Pdia2 mRNA is also proportionally reduced in hyperglycemic pancreas.

  19. Effects of cell-cycle-dependent expression on random fluctuations in protein levels.

    Science.gov (United States)

    Soltani, Mohammad; Singh, Abhyudai

    2016-12-01

    Expression of many genes varies as a cell transitions through different cell-cycle stages. How coupling between stochastic expression and cell cycle impacts cell-to-cell variability (noise) in the level of protein is not well understood. We analyse a model where a stable protein is synthesized in random bursts, and the frequency with which bursts occur varies within the cell cycle. Formulae quantifying the extent of fluctuations in the protein copy number are derived and decomposed into components arising from the cell cycle and stochastic processes. The latter stochastic component represents contributions from bursty expression and errors incurred during partitioning of molecules between daughter cells. These formulae reveal an interesting trade-off: cell-cycle dependencies that amplify the noise contribution from bursty expression also attenuate the contribution from partitioning errors. We investigate the existence of optimum strategies for coupling expression to the cell cycle that minimize the stochastic component. Intriguingly, results show that a zero production rate throughout the cell cycle, with expression only occurring just before cell division, minimizes noise from bursty expression for a fixed mean protein level. By contrast, the optimal strategy in the case of partitioning errors is to make the protein just after cell division. We provide examples of regulatory proteins that are expressed only towards the end of the cell cycle, and argue that such strategies enhance robustness of cell-cycle decisions to the intrinsic stochasticity of gene expression.

  20. Digestible fibre to starch ratio and protein level in diets for growing rabbits

    Directory of Open Access Journals (Sweden)

    Gerolamo Xiccato

    2010-01-01

    Full Text Available To evaluate the effect of digestible fibre (DF to starch ratio (0.8, 1.5, and 2.8 and protein level (15% and 16% on health status, digestive physiology, growth performance, and carcass traits, 246 rabbits weaned at 33 d were fed until slaughter (75 d six diets formulated according to a bi-factorial arrangement (3 DF to starch ratios by 2 protein levels. Growth performance and carcass quality at slaughter were not af- fected by treatments. Increasing DF to starch ratio did not modify dry matter digestibility (62.0% on average, while increased (P<0.001 DF digestibility (52.3 to 68.1%, stimulated caecal fermentation (total VFA: 56.0 vs 67.8 and 67.2 mmol/l; P=0.02 and changed VFA molar proportions. Increasing dietary protein increased digestibility of dry matter (P=0.02, crude protein (P<0.001 and digestible fibre (P<0.001 and increased cae- cal VFA production (P<0.01. The highest mortality (17.1% vs 1.5% average mortality of the other groups, P<0.001 was found in rabbits fed the diet with the lowest DF to starch ratio and the highest protein level.

  1. Insecticidal activity of residual Bt protein at the second trophic level

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Measurements were taken of Bt protein expressed in the leaves of transgenic cotton (Gossypium hirsutum) transformed with a synthesized Bt (Bacillus thuringiensis) cry1A gene and its persistent level in larval bodies and faeces of a non-targeted insect pest, beet armyworm (Spodoptera exigua). We performed enzyme linked immunosorbent assays (ELISA) and bioassays using neonate larvae of cotton bollworm (Helicoverpa armigera) to detect the insecticidal activity of residual Bt protein at the second trophic level. The results showed that Bt protein content in functional leaves was different at various developmental stages and was different among plants at the same stage. Even though Bt protein concentration in the larval bodies and faeces decreased 97.5%-99% compared to that found in cotton leaves subsequently fed to beet armyworm larvae, it still had a lethal effect on neonate cotton bollworm larvae. Therefore, Bt protein present at the second trophic level had insecticidal activity. This result is important in understanding and predicting the effect of transgenic plants on nontarget organisms.

  2. Retinol-binding protein-4 and hs-CRP levels in patients with migraine.

    Science.gov (United States)

    Tanik, Nermin; Celikbilek, Asuman; Metin, Aslı; Gocmen, Ayse Yesim; Inan, Levent Ertugrul

    2015-10-01

    Retinol-binding protein-4 (RBP4) and high-sensitivity C-reactive protein (hs-CRP) levels are associated with inflammation in patients with migraine. The release of proinflammatory cytokines during migraine results in recurrent sterile neurogenic inflammation. This study aimed to determine the correlation between RBP4 and hs-CRP levels, and migraine, which is considered an inflammatory disease. The study included 48 migraine patients and 40 age- and gender-matched controls. Migraine was diagnosed according to International Classification of Headache Disorders-II. The serum RBP4 level was measured using a commercial ELISA kit and hs-CRP was measured using an enzyme immunoassay test kit. The serum RBP4 level was significantly lower in the migraine patients than in the controls (P hs-CRP level was significantly higher in the migraine patients (P hs-CRP levels did not differ between the migraine patients with and without aura (P > 0.05). Migraine headache severity, frequency and duration were not correlated with serum RBP or hs-CRP levels (P > 0.05). The observed high hs-CRP level and low RBP4 level in migraine patients suggest that vitamin A might play a major role in the pathogenesis of migraine. It is known that inflammation is a key factor in many diseases. Additional research might result in a better understanding of the anti-inflammatory effects of vitamin A.

  3. Protein Fibrillar Nanopolymers: Molecular-Level Insights into Their Structural, Physical and Mechanical Properties

    Science.gov (United States)

    Trusova, Valeriya M.

    2015-09-01

    Amyloid fibrils represent a generic class of mechanically strong and stable biomaterials with extremely advantageous properties. Although amyloids were initially associated only with severe neurological disorders, the role of these structures nowadays is shifting from health debilitating to highly beneficial both in biomedical and technological aspects. Intensive involvement of fibrillar assemblies into the wide range of pathogenic and functional processes strongly necessitate the molecular level characterization of the structural, physical and elastic features of protein nanofibrils. In the present contribution, we made an attempt to highlight the up-to-date progress in the understanding of amyloid properties from the polymer physics standpoint. The fundamental insights into protein fibril behavior are essential not only for development of therapeutic strategies to combat the protein misfolding disorders but also for rational and precise design of novel biodegradable protein-based nanopolymers.

  4. Parasitosis gastrointestinal and hematocrit value in ewe fed with different levels of crude protein

    Directory of Open Access Journals (Sweden)

    Francisco de Assis Fonseca de Macedo

    2015-12-01

    Full Text Available We evaluated the parasitological behavior of sheep females fed with different levels of crude protein, by the value of hematocrit and egg counts per gram of feces (EPG in identifying severely infected animals. Were used 47 female sheep of Santa Ines, Texel and Ile de France, aged between eight and 12 months, wormed 30 days before the first data of crops, which were held every two weeks. Within each race, the animals were divided into two treatments, deferred by 12% or 16%, crude protein in the diet. It is obtained, laboratory, the value of hematocrit of each animal and the amount of EPG. The value of hematocrit showed a variation (28.5 a 34.0 throughout the experimental period, arising from changes (800 a 5400 of EPG values. No differences were identified for EPG and hematocrit levels between 12 and 16% of PB. Diet recommendation with 12% crude protein, by inferring lower commercial cost in feed.

  5. Variation of Serum Protein Level in Xiang Piglets at Different Month of Age

    Institute of Scientific and Technical Information of China (English)

    ChiYujie; GaoXuejung; 等

    1995-01-01

    Usual clinieal methods were used to determine the concentrations of serum protein,albumin and globulin in Xiang piglets from the lst month to the 6th month of age.Thirteen piglets(eight males and five females)born is Spring were selected from the Livestock Experimental Station of NEAU.The results showed that the serum protein level was different at different month of age.The lowest level was found at the 1st month of age,and the highest at the 4th month of age.The difference in serum protein concentration was significant between different months of age.There was ,however,no significant difference between sexes at the same month of age.

  6. Protein levels in enteral feeds: do these meet requirements in children with severe cerebral palsy?

    Science.gov (United States)

    Schoendorfer, Niikee; Tinggi, Ujang; Sharp, Nita; Boyd, Roslyn; Vitetta, Luis; Davies, Peter S W

    2012-05-01

    Children with cerebral palsy (CP) have been documented to have feeding difficulties, which increase in line with condition severity and result in lowered growth potential. Much nutrition literature surrounds energy intake and expenditure in these children, with less information available on other parameters such as protein and micronutrients, which are also important for growth and development. We examined differences in protein intake and a variety of protein metabolism indices in children with CP compared with controls. A total of twenty-four children aged 4-12 years with marked CP fed orally (O, n 15) or enterally (E, n 9) were recruited, including age-matched typically developing children (C, n 24). Fasting blood samples were analysed for levels of albumin, creatinine, urea and urate. Parents collected an exact food replica for three consecutive days of their child's actual intake, which were directly analysed for protein content. Significant differences were found in protein intakes between the groups (mean percentage minimum requirements: E = 178 (sd 47); O = 208 (sd 95); C = 311 (sd 119), P = 0·005). Despite all children consuming over recommended levels, children with CP had significantly reduced levels of the protein metabolic indices compared with controls. These include as z-scores: albumin mean C = 0·71 (sd 1·04) and CP = - 0·17 (sd 1·60), P = 0·03; creatinine C = - 2·06 (sd 0·46) and CP = - 3·11 (sd 0·98), P < 0·001; urate C = 0·18 (sd 0·62) and CP = - 0·58 (sd 0·93), P = 0·002. Post hoc analysis, the present data show potentially greater protein metabolism issues in enterally fed children, compared with the other groups. This may also support recent literature that suggests shortfalls in current recommendations.

  7. High-level expression of nonglycosylated human pancreatic lipase-related protein 2 in Pichia pastoris.

    Science.gov (United States)

    Sebban-Kreuzer, Corinne; Deprez-Beauclair, Paule; Berton, Amelie; Crenon, Isabelle

    2006-10-01

    The human pancreatic lipase-related protein 2 (HPLRP2) was produced in the methylotrophic yeast Pichia pastoris. The HPLRP2 cDNA corresponding to the protein coding sequence including the native signal sequence, was cloned into the pPIC9K vector and integrated into the genome of P. pastoris. P. pastoris transformants secreting high-level rHPLRP2 were obtained and the expression level into the liquid culture medium reached about 40mg/L after 4 days of culture. rHPLRP2 was purified by a single anion-exchange step after an overnight dialysis. N-terminal sequence analysis showed that the purified rHPLRP2 mature protein possessed a correct N-terminal amino acid sequence indicating that its signal peptide was properly processed. Mass spectrometry analysis showed that the recombinant HPLRP2 molecular weight was 52,532Da which was 2451Da greater than the mass calculated from the sequence of the protein (50,081Da) and 1536Da greater than the mass of the native human protein (50,996Da). In vitro deglycosylation experiments by peptide:N-glycosidase F (PNGase F) indicated that rHPLRP2 secreted from P. pastoris was N-glycosylated. Specific conditions were setup in order to obtain a recombinant protein free of glycan chain. We observed that blocking glycosylation in vivo by addition of tunicamycin in the culture medium during the production resulted in a correct processing of the rHPLRP2 mature protein. The lipase activity of glycosylated or nonglycosylated rHPLRP2, which was about 800U/mg on tributyrin, was inhibited by the presence of bile salts and not restored by adding colipase. In conclusion, the experimental procedure which we have developed will allow us to get a high-level production in P. pastoris of glycosylated and nonglycosylated rHPLRP2, suitable for subsequent biophysical and structural studies.

  8. CSF Neurofilament Proteins Levels are Elevated in Sporadic Creutzfeldt-Jakob Disease

    NARCIS (Netherlands)

    van Eijk, Jeroen J. J.; van Everbroeck, Bart; Abdo, W. Farid; Kremer, Berry P. H.; Verbeek, Marcel M.

    2010-01-01

    In this study we investigated the cerebrospinal fluid (CSF) levels of neurofilament light (NFL) and heavy chain (NFHp35), total tau (t-tau), and glial fibrillary acidic protein (GFAP) to detect disease specific profiles in sporadic Creutzfeldt Jakob disease (sCJD) patients and Alzheimer's disease

  9. CSF neurofilament proteins levels are elevated in sporadic Creutzfeldt-Jakob disease.

    NARCIS (Netherlands)

    Eijk, J.J.J. van; Everbroeck, B. van; Abdo, W.; Kremer, H.P.H.; Verbeek, M.M.

    2010-01-01

    In this study we investigated the cerebrospinal fluid (CSF) levels of neurofilament light (NFL) and heavy chain (NFHp35), total tau (t-tau), and glial fibrillary acidic protein (GFAP) to detect disease specific profiles in sporadic Creutzfeldt Jakob disease (sCJD) patients and Alzheimer's disease

  10. CSF Neurofilament Proteins Levels are Elevated in Sporadic Creutzfeldt-Jakob Disease

    NARCIS (Netherlands)

    van Eijk, Jeroen J. J.; van Everbroeck, Bart; Abdo, W. Farid; Kremer, Berry P. H.; Verbeek, Marcel M.

    2010-01-01

    In this study we investigated the cerebrospinal fluid (CSF) levels of neurofilament light (NFL) and heavy chain (NFHp35), total tau (t-tau), and glial fibrillary acidic protein (GFAP) to detect disease specific profiles in sporadic Creutzfeldt Jakob disease (sCJD) patients and Alzheimer's disease (A

  11. Effects of dietary protein and energy levels on cow manure extretion and ammonia votalization

    NARCIS (Netherlands)

    Stelt, van der B.; Vliet, van P.C.J.; Reijs, J.W.; Temminghoff, E.J.M.; Riemsdijk, van W.H.

    2008-01-01

    Adjusting dietary composition is considered an effective way to reduce nutrient losses to the environment. The effects of various dietary protein and energy levels on manure composition (Ca, Mg, K, Na, N, P, and pH) were studied by determining total and direct available (free) nutrient concentration

  12. Modeling of DNA and Protein Organization Levels with Cn3D Software

    Science.gov (United States)

    Stasinakis, Panagiotis K.; Nicolaou, Despoina

    2017-01-01

    The molecular structure of living organisms and the complex interactions amongst its components are the basis for the diversity observed at the macroscopic level. Proteins and nucleic acids are some of the major molecular components, and play a key role in several biological functions, such as those of development and evolution. This article…

  13. Long-term stability and circadian variation in circulating levels of surfactant protein D

    DEFF Research Database (Denmark)

    Hoegh, Silje Vermedal; Sorensen, Grith Lykke; Tornoe, Ida;

    2010-01-01

    Surfactant protein D (SP-D) is an oligomeric calcium-dependent lectin with important roles in innate host defence against infectious microorganisms. Several studies have shown that patients with inflammatory lung disease have elevated levels of circulating SP-D, and serum SP-D has been suggested ...

  14. C-reactive protein serum level in patients with psoriasis before and after treatment with narrow-band ultraviolet B*

    Science.gov (United States)

    Farshchian, Mahmoud; Ansar, Akram; Sobhan, Mohammadreza; Hoseinpoor, Valiollah

    2016-01-01

    Background C-reactive protein is an inflammatory biomarker and its level increases in the serum of psoriatic patients. Its level is also associated with Psoriasis Area and Severity Index score. Objective The aim of this study was to assess the decrement of serum C-reactive protein level with narrow-band ultraviolet B (NB-UVB) therapy. Methods C-reactive protein serum levels in psoriasis patients were measured before and after treatment with NB-UVB and the data were analyzed in relation to the Psoriasis Area and Severity Index score improvement. Results Baseline C-reactive protein levels among psoriatic patients were higher than normal. These levels decreased significantly after treatment (P<0.001). At the beginning of the study, patients with higher levels of C-reactive protein also had more extensive and severe skin involvement. The highest decrease in C-reactive protein was observed in patients who responded better to the treatment and achieved a higher Psoriasis Area and Severity Index 75%. There was an association between baseline Psoriasis Area and Severity Index scores and C-reactive protein levels. Conclusion Patients with moderate to severe plaque-type psoriasis had active systemic inflammation, which was demonstrated by increased levels of C-reactive protein. Furthermore, skin disease severity was correlated with C-reactive protein levels. Phototherapy healed the psoriatic skin lesions and reduced inflammation, while decreasing C-reactive protein levels. PMID:27828628

  15. The effects of high temperature level on square Bt protein concentration of Bt cotton

    Institute of Scientific and Technical Information of China (English)

    WANG Jun; CHEN Yuan; YAO Meng-hao; LI Yuan; WEN Yu-jin; ZHANG Xiang; CHEN De-hua

    2015-01-01

    Higher bol worm survival rates were detected after high temperature presented during square period in Bt cotton. The objective of this study was to investigate the effects of high temperature level on the Bt efifcacy of two different types of Bt cotton cultivars at squaring stage. During the 2011 to 2013 cotton growth seasons, high temperature treatments ranged from 34 to 44°C in climate chambers, and ifeld experiments under high temperature weather with various temperature levels were conducted to investigate the effects of the high temperature level on square Bt protein concentration and nitrogen metabolism. The climate chamber experiments showed that the square insecticidal protein contents reduced after 24 h elevated temperature treatments for both cultivars, whereas signiifcant declines of the square insecticidal protein contents were detected at temperature >38°C, and only slightly numerical reductions were observed when temperature below 38°C. Similar high temperature responses were also observed at the two ifeld experimental sites in 2013. Correspondingly, high temperature below 38°C seems have little effect on the square amino acid concentrations, soluble protein contents, glutam-ic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) activities as wel as protease and peptidase activities; however, when the temperature was above 38°C, reduced soluble protein contents, enhanced amino acid con-centrations, decreased GPT and GOT activities, bolstered protease and peptidase activities in square were detected. In general, the higher the temperature is (>38°C), the larger the changes for the above compound contents and key enzymes activities of the square protein cycle. The ifndings indicated that the unstable insect resistance of the square was related to high temperature level during square stage.

  16. A simple atomic-level hydrophobicity scale reveals protein interfacial structure.

    Science.gov (United States)

    Kapcha, Lauren H; Rossky, Peter J

    2014-01-23

    Many amino acid residue hydrophobicity scales have been created in an effort to better understand and rapidly characterize water-protein interactions based only on protein structure and sequence. There is surprisingly low consistency in the ranking of residue hydrophobicity between scales, and their ability to provide insightful characterization varies substantially across subject proteins. All current scales characterize hydrophobicity based on entire amino acid residue units. We introduce a simple binary but atomic-level hydrophobicity scale that allows for the classification of polar and non-polar moieties within single residues, including backbone atoms. This simple scale is first shown to capture the anticipated hydrophobic character for those whole residues that align in classification among most scales. Examination of a set of protein binding interfaces establishes good agreement between residue-based and atomic-level descriptions of hydrophobicity for five residues, while the remaining residues produce discrepancies. We then show that the atomistic scale properly classifies the hydrophobicity of functionally important regions where residue-based scales fail. To illustrate the utility of the new approach, we show that the atomic-level scale rationalizes the hydration of two hydrophobic pockets and the presence of a void in a third pocket within a single protein and that it appropriately classifies all of the functionally important hydrophilic sites within two otherwise hydrophobic pores. We suggest that an atomic level of detail is, in general, necessary for the reliable depiction of hydrophobicity for all protein surfaces. The present formulation can be implemented simply in a manner no more complex than current residue-based approaches.

  17. Protein-sparing effect with high energy levels and obtaining the optimum digestible energy/digestible protein ratio in growth diets to Oreochromis niloticus(L.

    Directory of Open Access Journals (Sweden)

    Felix Walter Gutierrez

    2014-03-01

    Full Text Available The sparing effect of the dietary protein using diets with high energy levels was evaluated. A factorial design was used to examine the interaction of two levels of protein (30 and 35% and two levels of digestible energy (3.3 and 3.7 kcal/g on the weight gain (WG, food conversion FC, retained protein (PR, retained energy (ER and protein efficiency ratio (PER in “sex reversal Nile tilapia” (Oreochromis niloticus. Diets contained anchovy and soybean meal as protein sources and yellow corn and wheat bran as energy sources. The fish oil was used to adjust the energy levels. There were highly significant interactions (P<0.01 between the experimental diets. In diets with 30% proteins, the increase of digestible energy significantly enhances WG, FC, PR, ER and PER, where the level of 3.7 kcal/g had the best performance. With 35% protein, the increase of digestible energy had not significant influence on the GP, CA, PR, ER and PER, however the growth of sex reversal Nile Tilapia was improved with the level of 3.3 kcal/g. When were compared both diets, were not find significant differences. Considering the protein cost, it can be concluded that best response was obtained with a diet containing 30 % of protein and 3.7 kcal/g of digestible energy and optimum digestible energy/ digestible protein ratio of 13.75 kcal/g protein.

  18. Microarray analysis of genes associated with cell surface NIS protein levels in breast cancer

    Directory of Open Access Journals (Sweden)

    Richardson Andrea L

    2011-10-01

    Full Text Available Abstract Background Na+/I- symporter (NIS-mediated iodide uptake allows radioiodine therapy for thyroid cancer. NIS is also expressed in breast tumors, raising potential for radionuclide therapy of breast cancer. However, NIS expression in most breast cancers is low and may not be sufficient for radionuclide therapy. We aimed to identify biomarkers associated with NIS expression such that mechanisms underlying NIS modulation in human breast tumors may be elucidated. Methods Published oligonucleotide microarray data within the National Center for Biotechnology Information Gene Expression Omnibus database were analyzed to identify gene expression tightly correlated with NIS mRNA level among human breast tumors. NIS immunostaining was performed in a tissue microarray composed of 28 human breast tumors which had corresponding oligonucleotide microarray data available for each tumor such that gene expression associated with cell surface NIS protein level could be identified. Results and Discussion NIS mRNA levels do not vary among breast tumors or when compared to normal breast tissues when detected by Affymetrix oligonucleotide microarray platforms. Cell surface NIS protein levels are much more variable than their corresponding NIS mRNA levels. Despite a limited number of breast tumors examined, our analysis identified cysteinyl-tRNA synthetase as a biomarker that is highly associated with cell surface NIS protein levels in the ER-positive breast cancer subtype. Conclusions Further investigation on genes associated with cell surface NIS protein levels within each breast cancer molecular subtype may lead to novel targets for selectively increasing NIS expression/function in a subset of breast cancers patients.

  19. Low Circulating Protein C Levels Are Associated with Lower Leg Ulcers in Patients with Diabetes

    Directory of Open Access Journals (Sweden)

    K. Whitmont

    2013-01-01

    Full Text Available Activated protein C (APC promotes angiogenesis and reepithelialisation and accelerates healing of diabetic ulcers. The aim of this study was to determine the relationship between the incidence of lower leg ulcers and plasma levels of APC's precursor, protein C (PC, in diabetic patients. Patients with diabetes who had a lower leg ulcer(s for >6 months (n=36 were compared with age-, type of diabetes-, and sex-matched subjects with diabetes but without an ulcer (n=36, controls. Total PC was assessed using a routine PC colorimetric assay. There was a significantly (P<0.001 lower level of plasma PC in patients with ulcers (103.3 ± 22.7, mean ± SD compared with control (127.1±34.0 subjects, when corrected for age and matched for gender and type of diabetes. Ulcer type (neuropathic, ischaemic, or mixed was not a significant covariate for plasma PC levels (P=0.35. There was no correlation between PC levels and gender, type of diabetes, HbA1c, or C-reactive protein in either group. In summary, decreased circulating PC levels are associated with, and may predispose to, lower leg ulceration in patients with diabetes.

  20. Association of MASP2 polymorphisms and protein levels with rheumatic fever and rheumatic heart disease.

    Science.gov (United States)

    Catarino, Sandra Jeremias dos Santos; Boldt, Angelica Beate Winter; Beltrame, Marcia Holsbach; Nisihara, Renato Mitsunori; Schafranski, Marcelo Derbli; de Messias-Reason, Iara Jose

    2014-12-01

    MASP-2 is a key protein of the lectin pathway of complement system. Several MASP2 polymorphisms were associated with MASP-2 serum levels or functional activity. Here we investigated a possible association between MASP2 polymorphisms and MASP-2 serum levels with the susceptibility to rheumatic fever (RF) and rheumatic heart disease (RHD). We haplotyped 11 MASP2 polymorphisms with multiplex sequence-specific PCR in 145 patients with history of RF from south Brazil (103 with RHD and 42 without cardiac lesion [RFo]) and 342 healthy controls. MASP-2 levels were determined by ELISA. The low MASP-2 producing p.377A and p.439H variants were negatively associated with RF (P=0.02, OR=0.36) and RHD (P=0.01, OR=0.25). In contrast, haplotypes that share the intron 9 - exon 12 g.1961795C, p.371D, p.377V and p.439R polymorphisms increased the susceptibility to RHD (P=0.02, OR=4.9). MASP-2 levels were associated with MASP2 haplotypes and were lower in patients (P<0.0001), which may reflect protein consumption due to complement activation. MASP2 gene polymorphisms and protein levels seem to play an important role in the development of RF and establishment of RHD.

  1. BDNF Val66Met polymorphism and protein levels in Amniotic Fluid

    Directory of Open Access Journals (Sweden)

    Calabrese Francesca

    2010-02-01

    Full Text Available Abstract Background Brain-Derived Neurotrophic Factor (BDNF is a neurotrophin which plays survival- and growth-promoting activity in neuronal cells and it is involved in cellular plasticity mechanisms as it controls activity dependent synaptic transmission. A functional polymorphism (Val66Met in the pro-region of BDNF, which affects the intracellular trafficking of proBDNF has been associated with memory and cognitive deficits as well as to an increased susceptibility for several psychiatric disorders especially those with a neurodevelopmental origin. To date, no study has evaluated the influence of the Val66Met polymorphism on BDNF levels in a peripheral system that may reflect fetal neurodevelopment. Therefore we investigated in amniotic fluids (AF obtained from 139 healthy women during 15-17 week of pregnancy, BDNF protein levels in correlation with the Val66Met polymorphism. Results Interestingly we found a significant BDNF protein levels reduction in 55 Met carriers (Val/Met and Met/Met (p = 0.002 as compared to 84 non carriers (Val/Val, and no effect of fetus gender, maternal age or gestation week on BDNF levels has been observed. Conclusion These results, although explorative, indicate that during fetal life the Val66Met genotype might influences BDNF protein levels in AF supporting the involvement of this polymorphism in behavioral and functional brain individual differences in the adulthood.

  2. Elevated C-Reactive Protein Levels, Psychological Distress, and Depression in 73 131 Individuals

    DEFF Research Database (Denmark)

    Wium-Andersen, Marie Kim; Ørsted, David Dynnes; Nielsen, Sune Fallgaard

    2013-01-01

    CONTEXT The pathogenesis of depression is not fully understood, but studies suggest that low-grade systemic inflammation contributes to the development of depression. OBJECTIVE To test whether elevated plasma levels of C-reactive protein (CRP) are associated with psychological distress and depres......CONTEXT The pathogenesis of depression is not fully understood, but studies suggest that low-grade systemic inflammation contributes to the development of depression. OBJECTIVE To test whether elevated plasma levels of C-reactive protein (CRP) are associated with psychological distress...... and depression. DESIGN We performed cross-sectional and prospective analyses of CRP levels in 4 clinically relevant categories using data from 2 general population studies. SETTING The Copenhagen General Population and the Copenhagen City Heart studies. PARTICIPANTS We examined 73 131 men and women aged 20...

  3. Increased levels of tau-like protein in patients with Down syndrome.

    Science.gov (United States)

    Mehta, P D; Patrick, B A; Dalton, A J; Aisen, P S; Emmerling, M E; Sersen, E A; Wisniewski, H M

    1999-11-19

    Tau-like protein levels from 40 Down syndrome (DS) persons (31-70 years old), 40 non-DS age-matched normal controls, 18 non-DS mentally retarded (MR) persons (26-91 years old), 25 probable Alzheimer disease (AD) patients (55-99 years old) and 24 non-demented elderly controls (54-79 years old) were measured using a sandwich enzyme linked immunosorbent assay. The levels were detected in 22 of 40 DS persons and were significantly higher in DS than any other group (P < 0.0001). There was no relationship between tau-like protein levels and age, gender or apolipoprotein E phenotypes in any of the five groups.

  4. Low levels of aflatoxin B1, ricin, and milk enhance recombinant protein production in mammalian cells.

    Directory of Open Access Journals (Sweden)

    Reuven Rasooly

    Full Text Available Gene expression in transduced mammalian cells correlates with virus titer, but high doses of vector for gene therapy leads to toxicity in humans and in animals. Changing the optimal tissue culture medium by adding low levels of environmental stressors, such as 1 µM of the fungal toxin aflatoxin B1 (AFB1, 1 ng of the castor bean protein toxin ricin, or 1% reconstituted milk, enhances transcription and increases production of proteins in transduced mammalian cells as demonstrated by production of the following three recombinant proteins: firefly luciferase, β-galactosidase, and green fluorescent protein (GFP. Higher concentrations of the stress-producing substances damage the cells beyond recovery, resulting in inhibited gene expression and cell death. We also evaluated the effect of the stressor substances on the enhanced infectivity of virus. The presented findings extend methods for large-scale transient recombinant protein production in mammalian cells and suggest that it may be possible to reduce the cytotoxicity of the adenovirus by reducing the virus titer without adversely affecting gene expression levels.

  5. Levels of digestible protein to surubim (Pseudoplatystoma sp. reared in net cages

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    Claucia Aparecida Honorato

    2014-10-01

    Full Text Available The Pseudoplatystoma sp. is species of carnivorous fish that require special attention in the diet offered. This work had the objective to determine the digestible protein requirement of juvenile the Pseudoplatystoma sp. reared in net cages. The test consisted of four isoenergetic diets (2606.69 ± 39.16 kcal kg-1 of digestible energy containing increasing levels of digestible protein (23, 24, 26 and 28%PD provided to juveniles of surubim (157.35±11.23g for five months. The parameters of growth, fillet composition, metabolic liver enzymes and morphometry of the intestine and liver were analyzed in completely randomized design with four treatments and four replicates. An increase of protein in the diet provided better weight gain. The metabolic liver enzymes increased in fish fed 24PD. The histopathological changes were not observed in the liver of the fish. The bowel histology showed adaptation to increased protein in the diet until the 26 level PD. juveniles of Pseudoplatystoma sp. Were demanding in digestible protein, showing the best results of production performance and nutrient use efficiency with the diet containing 28%PD.

  6. Comparison of C-reactive protein and high-sensitivity C-reactive protein levels in patients on hemodialysis

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    Imed Helal

    2012-01-01

    Full Text Available Chronic inflammation is highly prevalent in patients on hemodialysis (HD, as evidenced by increased levels of C-reactive protein (CRP. We compared CRP to high-sensitivity C-reactive protein (hs-CRP to determine whether it has any clinical implications and prognostic significance in terms of mortality. CRP was measured using a standard immunoturbidometric assay on the COBAS; INTEGRA system and hs-CRP was measured using the Dade Behring on the Konelab Nephelometer in 50 patients on HD. CRP (≥6 mg/L and hs-CRP (≥3 mg/L levels were elevated in 30% and 54% of the patients, respectively. A significant correlation was noted between hs-CRP and CRP levels (r = 0.98, P <0.001. Deming regression analysis showed that the slope was near one (r = 0.90; 0.83-0.94 and that the intercept was small. Multivariate regression confirmed that age above 40 years (RR = 3.69, P = 0.027 and duration on HD greater than five years (RR = 3.71, P = 0.028 remained significant independent predictors of serum hs-CRP. Thirteen patients died during follow-up (26%. Multivariate Cox regression demonstrated that hs-CRP (RR = 1.062, P = 0.03 and CRP levels (RR = 1.057, P = 0.009 and age (RR = 1.078, P = 0.001 were the most powerful predictors of mortality. The CRP standard assay presents a reasonable alternative to the hs-CRP assay in patients on HD. The advantages of the CRP standard assay are its online and real-time availability as well as lower costs, particularly in developing countries.

  7. Comparison of C-reactive protein and high-sensitivity C-reactive protein levels in patients on hemodialysis.

    Science.gov (United States)

    Helal, Imed; Zerelli, Lilia; Krid, Madiha; ElYounsi, Fethi; Ben Maiz, Hedi; Zouari, Bechir; Adelmoula, Jaouida; Kheder, Adel

    2012-05-01

    Chronic inflammation is highly prevalent in patients on hemodialysis (HD), as evidenced by increased levels of C-reactive protein (CRP). We compared CRP to high-sensitivity C-reactive protein (hs-CRP) to determine whether it has any clinical implications and prognostic significance in terms of mortality. CRP was measured using a standard immunoturbidometric assay on the COBAS® INTEGRA system and hs-CRP was measured using the Dade Behring on the Konelab Nephelometer in 50 patients on HD. CRP (≥6 mg/L) and hs-CRP (≥3 mg/L) levels were elevated in 30% and 54% of the patients, respectively. A significant correlation was noted between hs-CRP and CRP levels (r = 0.98, P <0.001). Deming regression analysis showed that the slope was near one (r = 0.90; 0.83-0.94) and that the intercept was small. Multivariate regression confirmed that age above 40 years (RR = 3.69, P = 0.027) and duration on HD greater than five years (RR = 3.71, P = 0.028) remained significant independent predictors of serum hs-CRP. Thirteen patients died during follow-up (26%). Multivariate Cox regression demonstrated that hs-CRP (RR = 1.062, P = 0.03) and CRP levels (RR = 1.057, P = 0.009) and age (RR = 1.078, P = 0.001) were the most powerful predictors of mortality. The CRP standard assay presents a reasonable alternative to the hs-CRP assay in patients on HD. The advantages of the CRP standard assay are its online and real-time availability as well as lower costs, particularly in developing countries.

  8. High-level expression of housefly cecropin A in Escherichia coli using a fusion protein

    Institute of Scientific and Technical Information of China (English)

    Xueli Zheng; Wei Wang

    2010-01-01

    Objective:To investigate the effect of utilizing a molecular partner on high-level expression of Musca domestica (M. domestica) cecropin in Escherichia coli (E. coli) and to identify the expressed products. Methods:The genomic sequence of M. domestica cecropin A (MC) and M. domestica ubiquitin (UBI) were searched from Genbank and amplified by reverse transcriptase polymerase chain reaction (RT-PCR). Two expression plasmids, pET32a-MC and pET32a-UBI-MC, were constructed and transferred into E. coli and were then induced by Isopropylβ-D-1-Thiogalactopyranoside (IPTG). The expression of the fusion proteins Trx-MC and Trx-UBI-MC was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Fusion protein Trx-MC was verified by Western blot analysis. The bactericidal activity of the purified MC was quantitatively determined using E. coli BL21(DE3). Results:The result showed that the fusion proteins were successively expressed in E. coli BL21 cells. A band at the expected position of 24 kDa representing the Trx-MC target protein was positivelystained, and the band at 4 kDa representing the hydrolysis of mature MC protein was also observed at the expected position. The expression levels of Trx-UBI-MC were higher than that of Trx-MC in E. coli. MC exhibited antimicrobial activity. Conclusions:With high-level expression of housefly cecropin A in E. coli using a fusion protein, MC exhibited antimicrobial activity.

  9. Alternatively Spliced Homologous Exons Have Ancient Origins and Are Highly Expressed at the Protein Level

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    Abascal, Federico; Ezkurdia, Iakes; Rodriguez-Rivas, Juan; Rodriguez, Jose Manuel; del Pozo, Angela; Vázquez, Jesús; Valencia, Alfonso; Tress, Michael L.

    2015-01-01

    Alternative splicing of messenger RNA can generate a wide variety of mature RNA transcripts, and these transcripts may produce protein isoforms with diverse cellular functions. While there is much supporting evidence for the expression of alternative transcripts, the same is not true for the alternatively spliced protein products. Large-scale mass spectroscopy experiments have identified evidence of alternative splicing at the protein level, but with conflicting results. Here we carried out a rigorous analysis of the peptide evidence from eight large-scale proteomics experiments to assess the scale of alternative splicing that is detectable by high-resolution mass spectroscopy. We find fewer splice events than would be expected: we identified peptides for almost 64% of human protein coding genes, but detected just 282 splice events. This data suggests that most genes have a single dominant isoform at the protein level. Many of the alternative isoforms that we could identify were only subtly different from the main splice isoform. Very few of the splice events identified at the protein level disrupted functional domains, in stark contrast to the two thirds of splice events annotated in the human genome that would lead to the loss or damage of functional domains. The most striking result was that more than 20% of the splice isoforms we identified were generated by substituting one homologous exon for another. This is significantly more than would be expected from the frequency of these events in the genome. These homologous exon substitution events were remarkably conserved—all the homologous exons we identified evolved over 460 million years ago—and eight of the fourteen tissue-specific splice isoforms we identified were generated from homologous exons. The combination of proteomics evidence, ancient origin and tissue-specific splicing indicates that isoforms generated from homologous exons may have important cellular roles. PMID:26061177

  10. Alternatively Spliced Homologous Exons Have Ancient Origins and Are Highly Expressed at the Protein Level.

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    Federico Abascal

    2015-06-01

    Full Text Available Alternative splicing of messenger RNA can generate a wide variety of mature RNA transcripts, and these transcripts may produce protein isoforms with diverse cellular functions. While there is much supporting evidence for the expression of alternative transcripts, the same is not true for the alternatively spliced protein products. Large-scale mass spectroscopy experiments have identified evidence of alternative splicing at the protein level, but with conflicting results. Here we carried out a rigorous analysis of the peptide evidence from eight large-scale proteomics experiments to assess the scale of alternative splicing that is detectable by high-resolution mass spectroscopy. We find fewer splice events than would be expected: we identified peptides for almost 64% of human protein coding genes, but detected just 282 splice events. This data suggests that most genes have a single dominant isoform at the protein level. Many of the alternative isoforms that we could identify were only subtly different from the main splice isoform. Very few of the splice events identified at the protein level disrupted functional domains, in stark contrast to the two thirds of splice events annotated in the human genome that would lead to the loss or damage of functional domains. The most striking result was that more than 20% of the splice isoforms we identified were generated by substituting one homologous exon for another. This is significantly more than would be expected from the frequency of these events in the genome. These homologous exon substitution events were remarkably conserved--all the homologous exons we identified evolved over 460 million years ago--and eight of the fourteen tissue-specific splice isoforms we identified were generated from homologous exons. The combination of proteomics evidence, ancient origin and tissue-specific splicing indicates that isoforms generated from homologous exons may have important cellular roles.

  11. Protein logic: a statistical mechanical study of signal integration at the single-molecule level.

    Science.gov (United States)

    de Ronde, Wiet; Rein ten Wolde, Pieter; Mugler, Andrew

    2012-09-05

    Information processing and decision-making is based upon logic operations, which in cellular networks has been well characterized at the level of transcription. In recent years, however, both experimentalists and theorists have begun to appreciate that cellular decision-making can also be performed at the level of a single protein, giving rise to the notion of protein logic. Here we systematically explore protein logic using a well-known statistical mechanical model. As an example system, we focus on receptors that bind either one or two ligands, and their associated dimers. Notably, we find that a single heterodimer can realize any of the 16 possible logic gates, including the XOR gate, by variation of biochemical parameters. We then introduce what to our knowledge is a novel idea: that a set of receptors with fixed parameters can encode functionally unique logic gates simply by forming different dimeric combinations. An exhaustive search reveals that the simplest set of receptors (two single-ligand receptors and one double-ligand receptor) can realize several different groups of three unique gates, a result for which the parametric analysis of single receptors and dimers provides a clear interpretation. Both results underscore the surprising functional freedom readily available to cells at the single-protein level. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  12. Stable Plastid Transformation for High-Level Recombinant Protein Expression: Promises and Challenges

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    Meili Gao

    2012-01-01

    Full Text Available Plants are a promising expression system for the production of recombinant proteins. However, low protein productivity remains a major obstacle that limits extensive commercialization of whole plant and plant cell bioproduction platform. Plastid genetic engineering offers several advantages, including high levels of transgenic expression, transgenic containment via maternal inheritance, and multigene expression in a single transformation event. In recent years, the development of optimized expression strategies has given a huge boost to the exploitation of plastids in molecular farming. The driving forces behind the high expression level of plastid bioreactors include codon optimization, promoters and UTRs, genotypic modifications, endogenous enhancer and regulatory elements, posttranslational modification, and proteolysis. Exciting progress of the high expression level has been made with the plastid-based production of two particularly important classes of pharmaceuticals: vaccine antigens, therapeutic proteins, and antibiotics and enzymes. Approaches to overcome and solve the associated challenges of this culture system that include low transformation frequencies, the formation of inclusion bodies, and purification of recombinant proteins will also be discussed.

  13. Effect of Diets with Different Energy and Protein Levels on Breast Muscle Characteristics at Broiler Chickens

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    Adela Marcu

    2013-05-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE In this paper was studied the effect of dietary energy and protein levels on breast muscle characteristics at broiler chickens, which were sacrificed at 42 days old. The genetic material was represented by broiler chickens that belonged to the „Ross-308” hybrid, with three groups (LC-control group, L1 and L2 experimental groups. In the growth periods (starter, growing and finishing have received compound feed ad libitum, with different energy and protein levels (LC-was conforming to recommendations of Aviagen Company; L1-higher with 10%; L2-lower with 10%. After evisceration, from each group were sampled breasts from 10 carcasses (five per sex and were determined: muscle mass, meat:bones ratio, chemical composition of meat, pH value (after evisceration up to 24 h of refrigeration and the thickness of myocytes in the superficial pectoral muscle. For these characteristics, highest values were obtained at L1 group, and the lowest values were at L2 group. At the L1 group, high levels of dietary proteins and energy has significantly influenced: muscle mass, meat:bones ratio, chemical composition of meat (water, proteins and lipids, pH value and the thickness of myocytes in the superficial pectoral muscle, as compared with LC and L2.

  14. Trace levels of innate immune response modulating impurities (IIRMIs) synergize to break tolerance to therapeutic proteins.

    Science.gov (United States)

    Verthelyi, Daniela; Wang, Vivian

    2010-12-22

    Therapeutic proteins such as monoclonal antibodies, replacement enzymes and toxins have significantly improved the therapeutic options for multiple diseases, including cancer and inflammatory diseases as well as enzyme deficiencies and inborn errors of metabolism. However, immune responses to these products are frequent and can seriously impact their safety and efficacy. Of the many factors that can impact protein immunogenicity, this study focuses on the role of innate immune response modulating impurities (IIRMIs) that could be present despite product purification and whether these impurities can synergize to facilitate an immunogenic response to therapeutic proteins. Using lipopolysaccharide (LPS) and CpG ODN as IIRMIs we showed that trace levels of these impurities synergized to induce IgM, IFNγ, TNFα and IL-6 expression. In vivo, trace levels of these impurities synergized to increase antigen-specific IgG antibodies to ovalbumin. Further, whereas mice treated with human erythropoietin showed a transient increase in hematocrit, those that received human erythropoietin containing low levels of IIRMIs had reduced response to erythropoietin after the 1(st) dose and developed long-lasting anemia following subsequent doses. This suggests that the presence of IIRMIs facilitated a breach in tolerance to the endogenous mouse erythropoietin. Overall, these studies indicate that the risk of enhancing immunogenicity should be considered when establishing acceptance limits of IIRMIs for therapeutic proteins.

  15. Trace levels of innate immune response modulating impurities (IIRMIs synergize to break tolerance to therapeutic proteins.

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    Daniela Verthelyi

    Full Text Available Therapeutic proteins such as monoclonal antibodies, replacement enzymes and toxins have significantly improved the therapeutic options for multiple diseases, including cancer and inflammatory diseases as well as enzyme deficiencies and inborn errors of metabolism. However, immune responses to these products are frequent and can seriously impact their safety and efficacy. Of the many factors that can impact protein immunogenicity, this study focuses on the role of innate immune response modulating impurities (IIRMIs that could be present despite product purification and whether these impurities can synergize to facilitate an immunogenic response to therapeutic proteins. Using lipopolysaccharide (LPS and CpG ODN as IIRMIs we showed that trace levels of these impurities synergized to induce IgM, IFNγ, TNFα and IL-6 expression. In vivo, trace levels of these impurities synergized to increase antigen-specific IgG antibodies to ovalbumin. Further, whereas mice treated with human erythropoietin showed a transient increase in hematocrit, those that received human erythropoietin containing low levels of IIRMIs had reduced response to erythropoietin after the 1(st dose and developed long-lasting anemia following subsequent doses. This suggests that the presence of IIRMIs facilitated a breach in tolerance to the endogenous mouse erythropoietin. Overall, these studies indicate that the risk of enhancing immunogenicity should be considered when establishing acceptance limits of IIRMIs for therapeutic proteins.

  16. Measures of bioavailable serum sex hormone levels in aging Chinese by protein chip

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yong; DU Guanhua; CHANG Shuying; MENG Xiaoluo; YU Huafeng; WANG Luning; HE Jinggui; ZHANG Baohe; ZHANG Juntian; GENG Meiyu

    2006-01-01

    The purpose of this study was to develop a protein chip technique based on receptor binding assays to measure bioavailable serum sex hormone levels (BSSHL). 224 aging healthy Chinese were investigated to get the referenced values of BSSHL for the first time. In the assays recombined sex hormone receptor proteins were jointed to polysaccharide coated slides to make protein chip, and the dose-dependence curves of sex hormone on chip were prepared. The data showed that this method had good precision (CV<16%) and accuracy (Bias<10%), and the sensitivity could reach 1 pmol/L. From the results, BSSHL of men and women declined with aging, but no significant differences were observed. The BSSHL of aging men were higher than those of women. The bioavailable serum androgen level of men was 52―112 pmol/L, women's was 3―70 pmol/L and the whole group was 41.9―81.4 pmol/L. The bioavailable serum estrogen level of men was 0.8―3.0 pmol/L, women's was 1.2―2.5 pmol/L and the whole group was 0.6―2.64 pmol/L. Based on the assays, BSSHL measurement by protein chip can meet the needs of epidemiological studies in terms of speed, accuracy and sample volume required, and was helpful in quantitative assessment of aging people's health.

  17. Normal human epidermal keratinocytes treated with 7-dehydrocholesterol express increased levels of heat shock protein.

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    Mammone, Thomas; Muizzuddin, Neelam; Goyarts, Earl; Gan, David; Giacomoni, Paolo; Marenus, Ken; Maes, Daniel

    2004-01-01

    Human skin, and its isolated cells, respond to insults with a variety of repair and protective mechanisms. One such mechanism is the production of heat shock proteins (HSPs). Heat shock proteins help the other cellular proteins fold correctly into their active three-dimensional structures. Therefore, they can enhance the survival of cells under harsh, denaturing conditions. In order to develop a means of promoting the heat shock response to prepare the skin to withstand insult, we are investigating materials that appear to protect the skin biologically. One such material is vitamin D3 and its precursors. We have observed that keratinocytes treated with 7-dehydrocholesterol (7-DHC), a precursor of vitamin D3, have increased levels of protein and mRNA for heat shock proteins. In addition, we observed that topically applied 7-DHC increases the minimal dose of UVB required to induce erythema. These data suggest that 7-DHC can induce heat shock proteins in skin keratinocytes and that they will be more resistant to UVB insult.

  18. Extraction of proteins with low fluoride level from Antarctic krill (Euphausia superba) and their composition analysis.

    Science.gov (United States)

    Wang, Lingzhao; Xue, Changhu; Wang, Yuming; Yang, Bao

    2011-06-08

    The extraction of proteins with low fluoride level (LFP) from Antarctic krill (Euphausia superba) was investigated in this work. The optimal conditions for protein solubilization were determined to be pH 11.5 and 4 °C. The proteins were solubilized two times; a water/krill ratio (mL/g) of 6 and a time of 30 min were used for the first step, whereas the second used a water/krill residue ratio (mL/g) of 3 and a time of 30 min. The optimum pH for protein precipitation was 4.6. A LFP with fluoride content of 9.86 mg/kg (dry weight) was finally obtained through a fluoride removal program. The protein yield of LFP was 52.68%. Composition analysis of LFP indicated it was composed of 66.96% of crude proteins (dry weight) and 33.01% of total lipids (dry weight),, and all nine essential amino acids were in sufficient amounts to meet FAO/WHO/UNU requirements for adults and infants. In addition, LFP could be taken as a good source of EPA and DHA for consideration of use as a food item for human consumption.

  19. Various dietary levels of protein and energy interaction on growth performance of white plumage japanese quails

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    Runjun Dowarah

    2014-06-01

    Full Text Available Aim: The present study was investigated to determine the optimum energy and protein requirement for growth performance and nutrient digestibility of white plumage Japanese quail during starter (1 to 3 week and finisher (4 to 5 week phases. Materials and Methods: Japanese quails were fed with nine diets consisting of three levels of energy (2600, 2800 and 3000; 2800, 2900 and 3000 Kcal ME/kg and protein (23, 25 and 27; 18, 20 and 22% CP during starter and finisher phases, respectively, in 3×3 factorial design. Results: Protein and energy individually had no influence on final body weight (FBW and weight gain, while feed efficiency was significantly (P<0.05 increased. The protein × energy interaction had significant (P<0.05 effect on growth and nutrient utilization during both starter and finisher phases. Conclusion: The growth performance was higher (P<0.05 in quails fed on high protein-high energy (HP- HE combination (27% CP and 3000 Kcal ME/kg and medium protein-high energy (MP-HE combination (20% CP and 3000 Kcal ME/kg during starter and finisher phase, respectively in autumn season. The diet with 20% CP and 3000 Kcal ME/kg significantly (P<0.05 increased nutrients digestibility and major minerals (Ca& P balance.

  20. Nutrient digestibility and growth of five breeds of sheep under different levels of undegradable protein

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    Yulistiani D

    2015-03-01

    Full Text Available A study was conducted to evaluate the effect of feeding different levels of rumen undegradable protein (UDP in concentrate on performance of five breeds of sheep. Namely: Compass Agrinak (CA; Garut Composite breed (KG; Barbados Cross breed (BC; St Croix breed (SC; and Local Garut (GL breed. Ten heads of sheep were used from each breed, in which each breed was grouped into 5 groups according to their body weight, and each sheep in each group was assigned to one of two treatments diet. The diet treatment consisted of concentrate containing different levels of undegradable protein (UDP. The levels of undegradable protein were 4.5% and 7.5%. Concentrate were formulated in iso nitrogen (CP content 16.7% and iso energy (ME content 2.5 Mcal/kg. During feeding trial the sheep were kept in individual pen for three months. Study was conducted in randomized complete block design in factorial 5 x 2 arrangement, 5 levels of breeds sheep and 2 levels of UDP content. Results shows that there was no interaction effect of breed and levels UDP on feed consumption, average daily gain and feed conversion. These variables were significantly (P0.05 by interaction between breed of sheep and UDP levels in the diet except for OM digestibility. While breed of sheep did not affect nutrient digestibility and UDP levels only affected NDF digestibility. From this study, it is concluded that increasing UDP in the diet did not improve growth performance of sheep. At similar quality of feed the growth performance was affected by breed of sheep in which KG and GL sheep had similar average daily gain (86.01 vs 82.38 g/day.

  1. Accounting for experimental noise reveals that mRNA levels, amplified by post-transcriptional processes, largely determine steady-state protein levels in yeast.

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    Gábor Csárdi

    2015-05-01

    Full Text Available Cells respond to their environment by modulating protein levels through mRNA transcription and post-transcriptional control. Modest observed correlations between global steady-state mRNA and protein measurements have been interpreted as evidence that mRNA levels determine roughly 40% of the variation in protein levels, indicating dominant post-transcriptional effects. However, the techniques underlying these conclusions, such as correlation and regression, yield biased results when data are noisy, missing systematically, and collinear---properties of mRNA and protein measurements---which motivated us to revisit this subject. Noise-robust analyses of 24 studies of budding yeast reveal that mRNA levels explain more than 85% of the variation in steady-state protein levels. Protein levels are not proportional to mRNA levels, but rise much more rapidly. Regulation of translation suffices to explain this nonlinear effect, revealing post-transcriptional amplification of, rather than competition with, transcriptional signals. These results substantially revise widely credited models of protein-level regulation, and introduce multiple noise-aware approaches essential for proper analysis of many biological phenomena.

  2. Serum C-reactive protein levels predict neurological outcome after aneurysmal subarachnoid hemorrhage

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    Flávio Ramalho Romero

    2012-03-01

    Full Text Available OBJECTIVES: Our aim was to evaluate the relationship between serum C-reactive protein (CRP levels and the neurological prognosis and development of vasospasm in patients with aneurysmal subarachnoid hemorrhage (aSAH. METHODS: Eighty-two adult patients with aSAH diagnoses were prospectively evaluated. Glasgow Coma Scale (GCS score, Hunt and Hess grade, Fisher grade, cranial CT scans, digital subtraction angiography studies and daily neurological examinations were recorded. Serial serum CRP measurements were obtained daily between admission and the tenth day. Glasgow Outcome Scale (GOS and the modified Rankin Scale (mRS were used to assess the prognosis. RESULTS: Serum CRP levels were related to severity of aSAH. Patients with lower GCS scores and higher Hunt and Hess and Fisher grades presented statistically significant higher serum CRP levels. Patients with higher serum CRP levels had a less favorable prognosis. CONCLUSIONS: Increased serum CRP levels were strongly associated with worse clinical prognosis in this study.

  3. The protein LJM 111 from Lutzomyia longipalpis salivary gland extract (SGE) accounts for the SGE-inhibitory effects upon inflammatory parameters in experimental arthritis model.

    Science.gov (United States)

    Grespan, Renata; Lemos, Henrique P; Carregaro, Vanessa; Verri, Waldiceu A; Souto, Fabricio O; de Oliveira, Carlo J F; Teixeira, Clarissa; Ribeiro, José Marcos; Valenzuela, Jesus G; Cunha, Fernando Q

    2012-04-01

    Several studies have pointed out the immunomodulatory properties of the Salivary Gland Extract (SGE) from Lutzomyia longipalpis. We aimed to identify the SGE component (s) responsible for its effect on ovalbumin (OVA)-induced neutrophil migration (NM) and to evaluate the effect of SGE and components in the antigen-induced arthritis (AIA) model. We tested the anti-arthritic activities of SGE and the recombinant LJM111 salivary protein (rLJM111) by measuring the mechanical hypernociception and the NM into synovial cavity. Furthermore, we measured IL-17, TNF-α and IFN-γ released by lymph nodes cells stimulated with mBSA or anti-CD3 using enzyme-linked immunosorbent assay (ELISA). Additionally, we tested the effect of SGE and rLJM111 on co-stimulatory molecules expression (MHC-II and CD-86) by flow cytometry, TNF-α and IL-10 production (ELISA) of bone marrow-derived dendritic cells (BMDCs) stimulated with LPS, chemotaxis and actin polymerization from neutrophils. Besides, the effect of SGE on CXCR2 and GRK-2 expression on neutrophils was investigated. We identified one plasmid expressing the protein LJM111 that prevented NM in OVA-challenged immunized mice. Furthermore, both SGE and rLJM111 inhibited NM and pain sensitivity in AIA and reduced IL-17, TNF-α and IFN-γ. SGE and rLJM111 also reduced MHC-II and CD-86 expression and TNF-α whereas increased IL-10 release by LPS-stimulated BMDCs. SGE, but not LJM 111, inhibited neutrophils chemotaxis and actin polymerization. Additionally, SGE reduced neutrophil CXCR2 expression and increased GRK-2. Thus, rLJM111 is partially responsible for SGE mechanisms by diminishing DC function and maturation but not chemoattraction of neutrophils. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Obese patients have higher circulating protein levels of dipeptidyl peptidase IV.

    Science.gov (United States)

    Stengel, Andreas; Goebel-Stengel, Miriam; Teuffel, Pauline; Hofmann, Tobias; Buße, Petra; Kobelt, Peter; Rose, Matthias; Klapp, Burghard F

    2014-11-01

    Dipeptidyl peptidase IV (DPPIV) is a protease with broad distribution involved in various homeostatic processes such as immune defense, psychoneuroendocrine functions and nutrition. While DPPIV protein levels were investigated in patients with hyporectic disorders, less is known under conditions of obesity. Therefore, we investigated DPPIV across a broad range of body mass index (BMI). Blood samples from hospitalized patients with normal weight (BMI 18.5-25 kg/m(2)), anorexia nervosa (BMI 50 kg/m(2), n = 15/group) were tested cross-sectionally and DPPIV concentration and total enzyme activity and the DPPIV targets, pancreatic polypeptide (PP) and glucagon-like peptide (GLP-1) were measured. DPPIV protein expression was detected in human plasma indicated by a strong band at the expected size of 110 kDa and another major band at 50 kDa, likely representing a fragment comprised of two heavy chains. Obese patients had higher DPPIV protein levels compared to normal weight and anorexics (+50%, p0.05), while the concentration/activity ratio was higher in obese patients (p<0.05). Plasma PP levels were highest in anorexic patients (∼ 2-fold increase compared to other groups, p<0.05), whereas GLP-1 did not differ among groups (p<0.05). Taken together, circulating DPPIV protein levels depend on body weight with increased levels in obese resulting in an increased concentration/activity ratio. Since DPPIV deactivates food intake-inhibitory hormones like PP, an increased DPPIV concentration/activity ratio might contribute to reduced food intake-inhibitory signaling under conditions of obesity.

  5. Individual variation in levels of haptoglobin-related protein in children from Gabon.

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    Heather J Imrie

    Full Text Available BACKGROUND: Haptoglobin related protein (Hpr is a key component of trypanosome lytic factors (TLF, a subset of high-density lipoproteins (HDL that form the first line of human defence against African trypanosomes. Hpr, like haptoglobin (Hp can bind to hemoglobin (Hb and it is the Hpr-Hb complexes which bind to these parasites allowing uptake of TLF. This unique form of innate immunity is primate-specific. To date, there have been no population studies of plasma levels of Hpr, particularly in relation to hemolysis and a high prevalence of ahaptoglobinemia as found in malaria endemic areas. METHODS AND PRINCIPAL FINDINGS: We developed a specific enzyme-linked immunosorbent assay to measure levels of plasma Hpr in Gabonese children sampled during a period of seasonal malaria transmission when acute phase responses (APR, malaria infection and associated hemolysis were prevalent. Median Hpr concentration was 0.28 mg/ml (range 0.03-1.1. This was 5-fold higher than that found in Caucasian children (0.049 mg/ml, range 0.002-0.26 with no evidence of an APR. A general linear model was used to investigate associations between Hpr levels, host polymorphisms, parasitological factors and the acute phase proteins, Hp, C-reactive protein (CRP and albumin. Levels of Hpr were associated with Hp genotype, decreased with age and were higher in females. Hpr concentration was strongly correlated with that of Hp, but not CRP. CONCLUSIONS/SIGNIFICANCE: Individual variation in Hpr levels was related to Hp level, Hp genotype, demographics, malaria status and the APR. The strong correlations between plasma levels of Hp and Hpr suggest that they are regulated by similar mechanisms. These population-based observations indicate that a more dynamic view of the relative roles of Hpr and Hpr-Hb complexes needs to be considered in understanding innate immunity to African trypanosomes and possibly other pathogens including the newly discovered Plasmodium spp of humans and

  6. Gallium nitrate regulates rat osteoblast expression of osteocalcin protein and mRNA levels.

    Science.gov (United States)

    Guidon, P T; Salvatori, R; Bockman, R S

    1993-01-01

    Gallium nitrate, a group IIIa metal salt, has been found to be clinically effective for the treatment of accelerated bone resorption in cancer-related hypercalcemia and Paget's disease. Here we report the effects of gallium nitrate on osteocalcin mRNA and protein levels on the rat osteoblast-like cell line ROS 17/2.8. Gallium nitrate reduced both constitutive and vitamin D3-stimulated osteocalcin protein levels in culture medium by one-half and osteocalcin mRNA levels to one-third to one-tenth of control. Gallium nitrate also inhibited vitamin D3 stimulation of osteocalcin and osteopontin mRNA levels but did not affect constitutive osteopontin mRNA levels. Among several different metals examined, gallium was unique in its ability to reduce osteocalcin mRNA levels without decreasing levels of other mRNAs synthesized by ROS 17/2.8 cells. The effects of gallium nitrate on osteocalcin mRNA and protein synthesis mimic those seen when ROS 17/2.8 cells are exposed to transforming growth factor beta 1 (TGF beta 1); however, TGF-beta 1 was not detected in gallium nitrate-treated ROS 17/2.8 cell media. Use of the RNA polymerase II inhibitor 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole demonstrated that gallium nitrate did not alter the stability of osteocalcin mRNA. Transient transfection assays using the rat osteocalcin promoter linked to the bacterial reporter gene chloramphenicol acetyltransferase indicated that gallium nitrate blocked reporter gene expression stimulated by the osteocalcin promoter. This is the first reported effect of gallium nitrate on isolated osteoblast cells.

  7. MSH3 polymorphisms and protein levels affect CAG repeat instability in Huntington's disease mice.

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    Stéphanie Tomé

    Full Text Available Expansions of trinucleotide CAG/CTG repeats in somatic tissues are thought to contribute to ongoing disease progression through an affected individual's life with Huntington's disease or myotonic dystrophy. Broad ranges of repeat instability arise between individuals with expanded repeats, suggesting the existence of modifiers of repeat instability. Mice with expanded CAG/CTG repeats show variable levels of instability depending upon mouse strain. However, to date the genetic modifiers underlying these differences have not been identified. We show that in liver and striatum the R6/1 Huntington's disease (HD (CAG∼100 transgene, when present in a congenic C57BL/6J (B6 background, incurred expansion-biased repeat mutations, whereas the repeat was stable in a congenic BALB/cByJ (CBy background. Reciprocal congenic mice revealed the Msh3 gene as the determinant for the differences in repeat instability. Expansion bias was observed in congenic mice homozygous for the B6 Msh3 gene on a CBy background, while the CAG tract was stabilized in congenics homozygous for the CBy Msh3 gene on a B6 background. The CAG stabilization was as dramatic as genetic deficiency of Msh2. The B6 and CBy Msh3 genes had identical promoters but differed in coding regions and showed strikingly different protein levels. B6 MSH3 variant protein is highly expressed and associated with CAG expansions, while the CBy MSH3 variant protein is expressed at barely detectable levels, associating with CAG stability. The DHFR protein, which is divergently transcribed from a promoter shared by the Msh3 gene, did not show varied levels between mouse strains. Thus, naturally occurring MSH3 protein polymorphisms are modifiers of CAG repeat instability, likely through variable MSH3 protein stability. Since evidence supports that somatic CAG instability is a modifier and predictor of disease, our data are consistent with the hypothesis that variable levels of CAG instability associated with

  8. MSH3 polymorphisms and protein levels affect CAG repeat instability in Huntington's disease mice.

    Science.gov (United States)

    Tomé, Stéphanie; Manley, Kevin; Simard, Jodie P; Clark, Greg W; Slean, Meghan M; Swami, Meera; Shelbourne, Peggy F; Tillier, Elisabeth R M; Monckton, Darren G; Messer, Anne; Pearson, Christopher E

    2013-01-01

    Expansions of trinucleotide CAG/CTG repeats in somatic tissues are thought to contribute to ongoing disease progression through an affected individual's life with Huntington's disease or myotonic dystrophy. Broad ranges of repeat instability arise between individuals with expanded repeats, suggesting the existence of modifiers of repeat instability. Mice with expanded CAG/CTG repeats show variable levels of instability depending upon mouse strain. However, to date the genetic modifiers underlying these differences have not been identified. We show that in liver and striatum the R6/1 Huntington's disease (HD) (CAG)∼100 transgene, when present in a congenic C57BL/6J (B6) background, incurred expansion-biased repeat mutations, whereas the repeat was stable in a congenic BALB/cByJ (CBy) background. Reciprocal congenic mice revealed the Msh3 gene as the determinant for the differences in repeat instability. Expansion bias was observed in congenic mice homozygous for the B6 Msh3 gene on a CBy background, while the CAG tract was stabilized in congenics homozygous for the CBy Msh3 gene on a B6 background. The CAG stabilization was as dramatic as genetic deficiency of Msh2. The B6 and CBy Msh3 genes had identical promoters but differed in coding regions and showed strikingly different protein levels. B6 MSH3 variant protein is highly expressed and associated with CAG expansions, while the CBy MSH3 variant protein is expressed at barely detectable levels, associating with CAG stability. The DHFR protein, which is divergently transcribed from a promoter shared by the Msh3 gene, did not show varied levels between mouse strains. Thus, naturally occurring MSH3 protein polymorphisms are modifiers of CAG repeat instability, likely through variable MSH3 protein stability. Since evidence supports that somatic CAG instability is a modifier and predictor of disease, our data are consistent with the hypothesis that variable levels of CAG instability associated with polymorphisms of

  9. Ultraviolet-ozone treatment reduces levels of disease-associated prion protein and prion infectivity

    Science.gov (United States)

    Johnson, C.J.; Gilbert, P.; McKenzie, D.; Pedersen, J.A.; Aiken, Judd M.

    2009-01-01

    Background. Transmissible spongiform encephalopathies (TSEs) are a group of fatal neurodegenerative diseases caused by novel infectious agents referred to as prions. Prions appear to be composed primarily, if not exclusively, of a misfolded isoform of the cellular prion protein. TSE infectivity is remarkably stable and can resist many aggressive decontamination procedures, increasing human, livestock and wildlife exposure to TSEs. Findings. We tested the hypothesis that UV-ozone treatment reduces levels of the pathogenic prion protein and inactivates the infectious agent. We found that UV-ozone treatment decreased the carbon and prion protein content in infected brain homogenate to levels undetectable by dry-ashing carbon analysis or immunoblotting, respectively. After 8 weeks of ashing, UV-ozone treatment reduced the infectious titer of treated material by a factor of at least 105. A small amount of infectivity, however, persisted despite UV-ozone treatment. When bound to either montmorillonite clay or quartz surfaces, PrPTSE was still susceptible to degradation by UV-ozone. Conclusion. Our findings strongly suggest that UV-ozone treatment can degrade pathogenic prion protein and inactivate prions, even when the agent is associated with surfaces. Using larger UV-ozone doses or combining UV-ozone treatment with other decontaminant methods may allow the sterilization of TSE-contaminated materials. ?? 2009 Aiken et al; licensee BioMed Central Ltd.

  10. Ultraviolet-ozone treatment reduces levels of disease-associated prion protein and prion infectivity

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    McKenzie Debbie

    2009-07-01

    Full Text Available Abstract Background Transmissible spongiform encephalopathies (TSEs are a group of fatal neurodegenerative diseases caused by novel infectious agents referred to as prions. Prions appear to be composed primarily, if not exclusively, of a misfolded isoform of the cellular prion protein. TSE infectivity is remarkably stable and can resist many aggressive decontamination procedures, increasing human, livestock and wildlife exposure to TSEs. Findings We tested the hypothesis that UV-ozone treatment reduces levels of the pathogenic prion protein and inactivates the infectious agent. We found that UV-ozone treatment decreased the carbon and prion protein content in infected brain homogenate to levels undetectable by dry-ashing carbon analysis or immunoblotting, respectively. After 8 weeks of ashing, UV-ozone treatment reduced the infectious titer of treated material by a factor of at least 105. A small amount of infectivity, however, persisted despite UV-ozone treatment. When bound to either montmorillonite clay or quartz surfaces, PrPTSE was still susceptible to degradation by UV-ozone. Conclusion Our findings strongly suggest that UV-ozone treatment can degrade pathogenic prion protein and inactivate prions, even when the agent is associated with surfaces. Using larger UV-ozone doses or combining UV-ozone treatment with other decontaminant methods may allow the sterilization of TSE-contaminated materials.

  11. Intake of Mung Bean Protein Isolate Reduces Plasma Triglyceride Level in Rats

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    Nobuhiko Tachibana

    2013-09-01

    Full Text Available ABSTRACTBackground: Mung bean is well known as a starch source, but the physiological effects of mung bean protein have received little attention. In this study, we isolated mung bean protein from de-starched mung bean solutions, and investigated its influence on lipid metabolism. Objective: The aim of this study is to clarify the influence of the lipid metabolism by consumption of mung bean protein isolate (MPIMethods: Diets containing either mung bean protein isolate (MPI or casein were fed to normal rats for 28 days.Results: Both groups ate the same amount of food, but the plasma triglyceride level, relative liver weight and liver lipid contents (cholesterol and triglyceride pool in the MPI group were significantly lower than in the casein group. In the MPI group, the expression of sterol regulatory-element binding factor 1 (SREBF1 mRNA in the liver was significantly different when compared with the casein group. The significantly lower levels of insulin and free fatty acids in the MPI-fed rats may be due to the regulation of genes related to lipid metabolism in the liver.Conclusions: These results suggest that MPI may improve the plasma lipid profile by normalizing insulin sensitivity.Keywords: mung bean, Vigna radiata L., 8S globulin, triglyceride, β-conglycinin, 7S globulin, insulin sensitivity, SREBF1

  12. COMPARISON OF SLAUGHTER, CARCASS VALUES OF SUDAN GOAT ECOTYPES FED DIFFERENT LEVELS OF ENERGY/PROTEIN

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    A.A. TAMEEM ELDAR

    2012-07-01

    Full Text Available This experiment aimed to assess the slaughter and carcass values of different Sudan goat ecotypes (Nubian, Desert and Swiss Nubian fed different levels of energy/protein. Nine male goat kids (2-3 months and average weight 9.23 kg of either ecotypes were used in a 3x3x3 arrangement, fed three experimental diets A (control, B and C, with varying energy: protein 1:0.14, 1:0.16 and 1:0.18 respectively. The study showed that Nubian goats possessed heavier slaughter weight; empty body weight, warm and cold carcass weight than the Desert and Swiss Nubian goats in the ration B and C. Dressing percentage on both basis (slaughter weight and empty body weight of Nubian were higher for ration B and C. Warm carcass weights of all goat ecotypes increased directly with energy: protein ratio. Highest weight of leg, loin, rack and neck and shoulder were recorded by Nubian goats for ration B and C. It was concluded that, all ecotype kids respond well to increasing energy protein levels. According to the results obtained local goat ecotypes, mainly Nubian goats, responded well to improved nutrition, therefore results obtained can be applied for the local goat ecotypes studied to encourage goat meat consumption and exportation.

  13. Effects of Dietary Crude Protein Levels and Cysteamine Supplementation on Protein Synthetic and Degradative Signaling in Skeletal Muscle of Finishing Pigs.

    Science.gov (United States)

    Zhou, Ping; Zhang, Lin; Li, Jiaolong; Luo, Yiqiu; Zhang, Bolin; Xing, Shen; Zhu, Yuping; Sun, Hui; Gao, Feng; Zhou, Guanghong

    2015-01-01

    Dietary protein levels and cysteamine (CS) supplementation can affect growth performance and protein metabolism of pigs. However, the influence of dietary protein intake on the growth response of CS-treated pigs is unclear, and the mechanisms involved in protein metabolism remain unknown. Hence, we investigated the interactions between dietary protein levels and CS supplementation and the effects of dietary crude protein levels and CS supplementation on protein synthetic and degradative signaling in skeletal muscle of finishing pigs. One hundred twenty barrows (65.84 ± 0.61 kg) were allocated to a 2 × 2 factorial arrangement with five replicates of six pigs each. The primary variations were dietary crude protein (CP) levels (14% or 10%) and CS supplemental levels (0 or 700 mg/kg). The low-protein (LP) diets (10% CP) were supplemented with enough essential amino acids (EAA) to meet the NRC AA requirements of pigs and maintain the balanced supply of eight EAA including lysine, methionine, threonine, tryptophan, valine, phenylalanine, isoleucine, and leucine. After 41 days, 10 pigs per treatment were slaughtered. We found that LP diets supplemented with EAA resulted in decreased concentrations of plasma somatostatin (SS) (Pdietary protein levels did not affect other traits. However, CS supplementation increased the average daily gain (Ppigs receiving CS. Additionally, CS supplementation increased the protein levels for the phosphorylated mammalian target of rapamycin (mTOR), eIF-4E binding protein 1, and ribosomal protein S6 kinase 1 (Pdietary protein levels and CS supplementation for all traits. In conclusion, dietary protein levels and CS supplementation influenced growth and protein metabolism through independent mechanisms in pigs. In addition, LP diets supplemented with EAA did not affect growth performance and other traits except the concentrations of SS and PUN probably through maintenance of protein synthesis and degradation signaling. Moreover, CS

  14. Relationship between the plasma levels of neurodegenerative proteins and motor subtypes of Parkinson's disease.

    Science.gov (United States)

    Ding, Jian; Zhang, Jiejin; Wang, Xixi; Zhang, Li; Jiang, Siming; Yuan, Yongsheng; Li, Junyi; Zhu, Lin; Zhang, Kezhong

    2017-03-01

    The aim of our study is to examine the plasma levels of the four kinds of neurodegenerative proteins in plasma: α-syn, T-tau, P-tau181, and Aβ-42 in Parkinson's disease (PD) and to evaluate the relationship between their plasma levels and PD motor subtypes. 84 patients with PD were enrolled in our study, and finally, 73 of them were classified into the tremor-dominant subtype (TD) and the postural instability gait difficulty subtype (PIGD). Their motor performance was evaluated by a series of clinical assessments: Freezing of Gait Questionnaire (FOGQ), Timed Up and Go (TUGs), Tinetti balance, and Tinetti gait. Plasma levels of these proteins were measured by enzyme-linked immunosorbent assay (ELISA). The plasma level of α-syn was significantly higher in PD patients when compared to controls (p = 0.004), and significantly higher in the PIGD group when compared to the TD group (p = 0.03). While the plasma level of Aβ-42 was significantly lower in PD patients than in controls (p = 0.002), and significantly lower in the PIGD group than in the TD group (p = 0.05). In PD patients, the plasma level of α-syn (r = -0.355, p score, even after performing multiple linear regression (p = 0.002). While the plasma level of Aβ-42 (r = -0.261, p score and remained correlate when performed multiple linear regression (p = 0.005). The patients with PIGD subtype are characterized with a lower level of plasma Aβ-42 and a higher plasma level of α-syn, which may be used as biomarkers for diagnosis and progression of the subtypes of PD.

  15. Serum Adenosine deaminase activity and C-reactive protein levels in unstable angina

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    Rani Surekha

    2003-01-01

    Full Text Available In unstable angina (USA patients, immunological responses contributing to inflammation play a vital role in plaque rupture and thrombosis causing stroke. In the present study an attempt is made to estimate the levels of adenosine deaminase activity, an immunoenzyme marker and C-reactive protein, a marker of inflammation in USA patients. 45 patients presenting USA and 50 age and sex matched healthy controls were included in the study. Serum ADA activity was measured spectrophotometrically at 630nm and serum C-reactive protein was detected using Avitex CRP kit, which is a rapid latex agglutination test. The Mean ADA levels were 41.15 ± 11.04 in patients and 20.71±5.63 in controls and 66.6% of patients and none of the controls were positive to CRP. The present study observed the importance of ADA as a serum marker in addition to CRP for assessing the immune response in USA patients.

  16. Blood parameters in growing pigs fed increasing levels of bacterial protein meal

    DEFF Research Database (Denmark)

    Hellwing, Anne Louise Frydendahl; Tauson, Anne-Helene; Skrede, Anders

    2007-01-01

    The experiment investigated the effects of increasing dietary levels of bacterial protein meal (BPM) on various blood parameters reflecting protein and fat metabolism, liver function, and purine base metabolism in growing pigs. Sixteen barrows were allocated to four different experimental diets....... The control diet was based on soybean meal. In the other three diets soybean meal was replaced with increasing levels of BPM, approximately 17%, 35%, and 50% of the nitrogen being derived from BPM. Blood samples from the jugular vein were taken when the body weights of the pigs were approximately 10 kg, 21 kg......, 45 kg, and 77 kg. The blood parameters reflecting fat metabolism and liver funtion were not affected by diet. Both the plasma albumin and uric acid concentrations tended to decrease (P = 0.07 and 0.01, respectively) with increasing dietary BPM content, whereas the plasma glucose concentration tended...

  17. The serum protein carbonyl content level in relation to exercise stress test

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    Titiporn Mekrungruangwong

    2012-01-01

    Full Text Available Background: Protein carbonyl (P is oxidatively-modified protein with diagnostic potential for acute myocardial infarction. However, many findings indicated the elevation of serum PC content level related to exercise, which could cause false positive results and limiting the specificity for acute coronary syndrome diagnosis. This study aims to evaluate the level of serum protein carbonyl content in healthy volunteers subjected to exercise stress test (EST. Materials and Methods: Serum from healthy volunteers was collected 5-10 min before performing EST and 1 hour after the EST was achieved. The serum was collected, and the serum PC content level was determined by spectrophotometric DNPH assay. Results: The serum PC content level after exercise stress test was significantly higher than that of before performing EST (0.373 ± 0.05 nM/mg vs. 0.275 ± 0.02 nM/mg, P < 0.0001. The results demonstrated that in both male and female, serum PC content level after EST was significantly higher than that of before performing EST (0.29 ± 0.03 nM/mg vs. 0.36 ± 0.05 nM/mg P < 0.0001 in male, 0.27 ± 0.02 nM/mg vs. 0.38 ± 0.06 nM/mg P < 0.0001 in female, respectively. Conclusions: This study demonstrated that exercise stress test could result in non-specificity and false positive increasing in serum PC content level in healthy subjects, which may cause misinterpretation when using PC as cardiac marker, especially in patients, who underwent exercise stress test or patients who performing heavy physical activities.

  18. Patients with nephrolithiasis had lower fetuin-A protein level in urine and renal tissue.

    Science.gov (United States)

    Wu, Yong Xian; Li, Cheng Yang; Deng, Yao Liang

    2014-02-01

    Fetuin-A acts as an inhibitor of systemic and local ectopic calcification and inflammatory response, but the role of fetuin-A in the etiology of urolithiasis is still unclear. We aim to investigate the expression of fetuin-A in the serum, urine and renal tissue of patients with or without nephrolithiasis. 48 patients with nephrolithiasis (group A) and 32 individuals without urolithiasis (group B, control group) were enrolled into our study. Level of fetuin-A in serum and urine was measured by ELISA, and expression of fetuin-A in renal tissue was localized and assessed by immunohistochemistry, real-time polymerase chain reaction, and Western blotting, respectively. Indexes of oxidative stress in kidney were evaluated. Other routine serum and urine chemistries for inpatients were measured biochemically. The results showed that fetuin-A expressed widely in the proximal and distal renal tubule, the thin segment of Henle's loop and the collecting duct epithelium. There were no differences in serum fetuin-A level between the two groups. Compared with control group, cellular expression of P47phox and fetuin-A mRNAs in the renal tissue of patients with nephrolithiasis increased, the level of MDA in renal tissue and the level of urinary calcium also increased, but urinary and renal fetuin-A protein and the activities of SOD in renal tissue decreased. Correlation analysis showed that there was a negative correlation between the level of renal fetuin-A protein and the expression of P47phox mRNA and MDA. These results revealed that nephrolithiasis patients had lower fetuin-A protein level in urine and renal tissue.

  19. Effect of physical training on glucose transporter protein and mRNA levels in rat adipocytes

    DEFF Research Database (Denmark)

    Stallknecht, B; Andersen, P H; Vinten, J

    1993-01-01

    Physical training increases insulin-stimulated glucose transport and the number of glucose transporters in adipocytes measured by cytochalasin B binding. In the present study we used immunoblotting to measure the abundance of two glucose transporters (GLUT-4, GLUT-1) in white adipocytes from....../or intrinsic activity). GLUT-1 protein and mRNA levels/adipocyte volume did not change with age or training....

  20. Serum levels of C-reactive protein in adolescents with periodontitis

    DEFF Research Database (Denmark)

    López, Rodrigo; Baelum, Vibeke; Hedegaard, Chris Juul;

    2011-01-01

    Background: The results of several cross-sectional studies suggested a relationship between periodontitis and higher serum levels of C-reactive protein (CRP). Most of these studies were restricted to adult study groups with severe periodontal inflammation, and the potential effects of confounding...... ng/ml (31 to 183 ng/ml), respectively (P = 0.8). Conclusions: Serum levels of CRP were not significantly higher among subjects with periodontitis than among controls. However, a statistically significant positive association between percentages of sites with bleeding on probing and log...

  1. Small intestine development of laying hens fed different fiber sources diets and crude protein levels

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    MFFM Praes

    2011-09-01

    Full Text Available The objective of the presente study was to evaluate the effects on different dietary fiber sources and crude protein levels on the intestinal morphometry of commercial layers. Isa Brown® layers with 48 weeks of age were distributed in a completely randomized experimental design with a 3 x 2 + 1 factorial arrangement, resulting in seven treatments with seven replicates of eight birds each. At the end of the fourth experimental period (28 days each, birds were 64 weeks of age and were randomly chosen (two birds per replicate, totaling 14 birds per treatment, weighed and sacrificed by neck dislocation. Their intestine was dissected and the duodenum, jejunum and ileum were collected for subsequent analysis of intestinal morphometry. Treatments consisted of diets containing three different fiber sources (cottonseed hulls, soybean hulls or rice husks and two crude protein levels (12% or 16%. Soybean hulls and 16% crude protein level promoted, in general, an increase in villus height and crypt depth in the three intestinal segments. In the duodenum, the control diet resulted in higher villus height and crypt depth relative to the diets containing fiber. In the jejunum, higher crypt depth values. In the ileum, dietary fiber increased villus height as compared to the control diet.

  2. Melamine Impairs Female Fertility via Suppressing Protein Level of Juno in Mouse Eggs.

    Science.gov (United States)

    Dai, Xiaoxin; Zhang, Mianqun; Lu, Yajuan; Miao, Yilong; Zhou, Changyin; Sun, Shaochen; Xiong, Bo

    2015-01-01

    Melamine is an organic nitrogenous compound widely used as an industrial chemical, and it has been recently reported by us that melamine has a toxic effect on the female reproductive system in mice, and renders females subfertile; the molecular basis, however, has not been adequately assessed. In the present study, we explore the underlying mechanism regarding how melamine compromises fertility in the mouse. The data showed that melamine exposure significantly impaired the fertilization capability of the egg during in vitro fertilization. To further figure out the cause, we analyzed ovastacin localization and protein level, the sperm binding ability of zona pellucida, and ZP2 cleavage status in unfertilized eggs from melamine fed mice, and no obvious differences were found between control and treatment groups. However, the protein level of Juno on the egg plasma membrane in the high-dose feeding group indeed significantly decreased compared to the control group. Thus, these data suggest that melamine compromises female fertility via suppressing Juno protein level on the egg membrane.

  3. Inclusion of Yucca schidigera extract in diets with different protein levels for dogs.

    Science.gov (United States)

    Dos Reis, Jéssica S; Zangerônimo, Márcio G; Ogoshi, Rosana C S; França, Janine; Costa, Adriano C; Almeida, Thomás N; Dos Santos, João P F; Pires, Carolina P; Chizzotti, Ana F; Leite, Carlos A L; Saad, Flávia M O B

    2016-08-01

    This study evaluated the effects of inclusion of Yucca schidigera extract (YSE) in two diets with different levels of crude protein (CP) for dogs on facal odour, nutrient digestibility, ammonia concentration in feces and hematological and serum biochemical profiles. Twenty adults Beagles were used, distributed in a randomized block design in a 2 × 4 factorial design (two diets, 25% and 34% CP, and four YSE levels: 0, 250, 500 and 750 mg/kg) with five replicates, obtained during two experimental periods. The fecal odour reduced (P < 0.05) when 500 mg/kg of YSE was used in diets with higher CP. The inclusion of YSE reduced (P < 0.05) fecal ammonia, and the inclusion of 250 and 500 mg/kg YSE reduced intestinal gas. The inclusion of 750 mg/kg YSE increased the mean corpuscular hemoglobin (MCH), alanine aminotransferase (ALT) activity and tended to increase the serum cholesterol concentration, regardless of the protein level of the diets. There was no effect on the digestibility of nutrients, fecal consistency, nitrogen balance and thickness of the intestinal wall. The inclusion of 500 mg/kg YSE is effective in reducing fecal odour in dogs receiving diets with 34% of CP. Regardless of the protein content, YSE reduces fecal ammonia, but may cause adverse effects if included at higher doses. © 2016 Japanese Society of Animal Science.

  4. Increased Levels of Antinutritional and/or Defense Proteins Reduced the Protein Quality of a Disease-Resistant Soybean Cultivar

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    Daniele O. B. Sousa

    2015-07-01

    Full Text Available The biochemical and nutritional attributes of two soybean (Glycine max (L. Merr. cultivars, one susceptible (Seridó and the other resistant (Seridó-RCH to stem canker, were examined to assess whether the resistance to pathogens was related to levels of antinutritional and/or defense proteins in the plant and subsequently affected the nutritional quality. Lectin, urease, trypsin inhibitor, peroxidase and chitinase activities were higher in the resistant cultivar. Growing rats were fed with isocaloric and isoproteic diets prepared with defatted raw soybean meals. Those on the Seridó-RCH diet showed the worst performance in terms of protein quality indicators. Based on regression analysis, lectin, trypsin inhibitor, peroxidase and chitinase appear to be involved in the resistance trait but also in the poorer nutritional quality of Seridó-RCH. Thus, the development of cultivars for disease resistance may lead to higher concentrations of antinutritional compounds, affecting the quality of soybean seeds. Further research that includes the assessment of more cultivars/genotypes is needed.

  5. Wilm's tumor-1 protein levels in urinary exosomes from diabetic patients with or without proteinuria.

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    Anuradha Kalani

    Full Text Available BACKGROUND: Podocyte injury is an early feature of diabetic nephropathy (DN. Recently, urinary exosomal Wilm's tumor-1 protein (WT1, shed by renal epithelial cells, has been proposed as a novel biomarker for podocyte injury. However, its usefulness as biomarker for early diabetic nephropathy has not been verified yet. We investigated urinary exosomal WT1 in type-1 diabetic patients to confirm its role as a non-invasive biomarker for predicting early renal function decline. METHODS: The expression of WT1 protein in urinary exosomes from spot urine samples of type-1 diabetes mellitus patients (n = 48 and healthy controls (n = 25 were analyzed. Patients were divided based on their urinary albumin excretion, ACR (mg/g creatinine into non- proteinuria group (ACR30 mg/g, n = 18. Regression analysis was used to assess the association between urinary exosomal levels of WT1 with parameters for renal function. Receiver Operating Characteristic (ROC curve analysis was used to determine the diagnostic performance of exosomal WT-1. RESULTS: WT1 protein was detected in 33 out of 48 diabetic patients and in only 1 healthy control. The levels of urinary exosomal WT1 protein is significantly higher (p = 0.001 in patients with proteinuria than in those without proteinuria. In addition, all the patients with proteinuria but only half of the patients without proteinuria were positive for exosomal WT1. We found that the level of exosomal WT1 were associated with a significant increase in urine protein-to-creatinine ratio, albumin-to-creatinine ratio, and serum creatinine as well as a decline in eGFR. Furthermore, patients exhibiting WT1-positive urinary exosomes had decreased renal function compared to WT1-negative patients. ROC analysis shows that WT-1 effectively predict GFR<60 ml. min-1/1.73 m(2. CONCLUSION: The predominant presence of WT1 protein in urinary exosomes of diabetic patients and increase in its expression level with decline in renal

  6. Myelin basic protein and ischemia modified albumin levels in acute ischemic stroke cases.

    Science.gov (United States)

    Can, Serdar; Akdur, Okhan; Yildirim, Ahmet; Adam, Gurhan; Cakir, Dilek Ulker; Karaman, Handan Isin Ozisik

    2015-01-01

    To investigate early diagnostic effects of serum myelin basic protein (MBP) and ischemic modified albumin (IMA) levels in patients with ischemic stroke. Fifty patients who presented to an emergency service with acute ischemic stroke between June 2013 to March 2014 were evaluated with the National Institute of Health Stroke Scale (NIHSS) and diffusion-weighted magnetic resonance imaging (MRI). Thirty four healthy cases were included as control group. All patients' serum IMA and MBP level were assessed. Mean IMA value was 0.52±0.25 cases with acute ischemic stroke and serum IMA levels were significantly higher than the control group (pacute ischemic stroke group and control group related to the MBP serum levels (P>0.05). Statistically significant correlation was detected between the volumes of diffusion restriction on MRI and NIHSS score (P=0.002, r=0.43) and IMA (P=0.015, r=0.344) levels. We have found that serum IMA levels are elevated in acute ischemic stroke cases and these levels are correlated with the ischemic tissue volume. MBP levels do not increase in early period of stroke cases.

  7. Correlation of MGMT promoter methylation status with gene and protein expression levels in glioblastoma

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    Miyuki Uno

    2011-01-01

    Full Text Available OBJECTIVES: 1 To correlate the methylation status of the O6-methylguanine-DNA-methyltransferase (MGMT promoter to its gene and protein expression levels in glioblastoma and 2 to determine the most reliable method for using MGMT to predict the response to adjuvant therapy in patients with glioblastoma. BACKGROUND: The MGMT gene is epigenetically silenced by promoter hypermethylation in gliomas, and this modification has emerged as a relevant predictor of therapeutic response. METHODS: Fifty-one cases of glioblastoma were analyzed for MGMT promoter methylation by methylation-specific PCR and pyrosequencing, gene expression by real time polymerase chain reaction, and protein expression by immunohistochemistry. RESULTS: MGMT promoter methylation was found in 43.1% of glioblastoma by methylation-specific PCR and 38.8% by pyrosequencing. A low level of MGMT gene expression was correlated with positive MGMT promoter methylation (p = 0.001. However, no correlation was found between promoter methylation and MGMT protein expression (p = 0.297. The mean survival time of glioblastoma patients submitted to adjuvant therapy was significantly higher among patients with MGMT promoter methylation (log rank = 0.025 by methylation-specific PCR and 0.004 by pyrosequencing, and methylation was an independent predictive factor that was associated with improved prognosis by multivariate analysis. DISCUSSION AND CONCLUSION: MGMT promoter methylation status was a more reliable predictor of susceptibility to adjuvant therapy and prognosis of glioblastoma than were MGMT protein or gene expression levels. Methylation-specific polymerase chain reaction and pyrosequencing methods were both sensitive methods for determining MGMT promoter methylation status using DNA extracted from frozen tissue.

  8. Determination of Moisture Levels, Protein and Water Absorption of Chicken Giblets

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    D Scaratti

    2016-03-01

    Full Text Available Abstract This study aimed at evaluating the levels of moisture, protein, water to protein ratio, and water absorption during chilling of chicken giblets (heart, liver, and gizzard to set legal limits of water absorption during this process. The survey was conducted in the southern Brazil, the largest broiler-producing region of this country. Giblets (heart, liver, and gizzard were collected fresh from the processing line after evisceration and at the exit of the chiller after the immersion process from two processing plants. One of the plants (PP1 processes small chickens (1,100g live weight and PP2 processes chickens with 2,800g live weight. In total, 448 samples were collected. Laboratory tests were performed in duplicate for each parameter measured. The results show that moisture levels of fresh giblets were higher in the gizzard, followed by the liver and the heart, whereas in chilled giblets, the gizzard still maintained the highest moisture level, but was followed by the heart and then the liver. Both in fresh and chilled samples, the liver presented the highest protein content, followed by the gizzard and the heart. Water to protein ratios were higher in chilled than in fresh samples, and was highest in the heart, followed by the gizzard and the liver. After immersion in the chiller, the heart presented the highest water absorption rate (6.59%, which was significantly higher compared with those of the liver (4.16% and the gizzard (4.51%. Considering that the water absorption rates obtained both in fresh and chilled chicken giblets were below 8.00%, the following upper limits of water absorption are suggested for chicken giblet processing in Brazil: 7.0% for the heart, and 5.0% for the gizzard and the liver.

  9. Unraveling protein folding mechanism by analyzing the hierarchy of models with increasing level of detail

    Science.gov (United States)

    Hayashi, Tomohiko; Yasuda, Satoshi; Škrbić, Tatjana; Giacometti, Achille; Kinoshita, Masahiro

    2017-09-01

    Taking protein G with 56 residues for a case study, we investigate the mechanism of protein folding. In addition to its native structure possessing α-helix and β-sheet contents of 27% and 39%, respectively, we construct a number of misfolded decoys with a wide variety of α-helix and β-sheet contents. We then consider a hierarchy of 8 different models with increasing level of detail in terms of the number of entropic and energetic physical factors incorporated. The polyatomic structure is always taken into account, but the side chains are removed in half of the models. The solvent is formed by either neutral hard spheres or water molecules. Protein intramolecular hydrogen bonds (H-bonds) and protein-solvent H-bonds (the latter is present only in water) are accounted for or not, depending on the model considered. We then apply a physics-based free-energy function (FEF) corresponding to each model and investigate which structures are most stabilized. This special approach taken on a step-by-step basis enables us to clarify the role of each physical factor in contributing to the structural stability and separately elucidate its effect. Depending on the model employed, significantly different structures such as very compact configurations with no secondary structures and configurations of associated α-helices are optimally stabilized. The native structure can be identified as that with lowest FEF only when the most detailed model is employed. This result is significant for at least the two reasons: The most detailed model considered here is able to capture the fundamental aspects of protein folding notwithstanding its simplicity; and it is shown that the native structure is stabilized by a complex interplay of minimal multiple factors that must be all included in the description. In the absence of even a single of these factors, the protein is likely to be driven towards a different, more stable state.

  10. Serum levels of hypersensitive-C-reactive protein in moderate and severe acne

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    M R Namazi

    2015-01-01

    Full Text Available Background: Elevation of C-reactive protein (CRP has been reported to occur in psoriasis, urticaria, acne, rosacea and many other dermatological and nondermatological conditions. Chronic systemic inflammation has been implicated in the development of neuropsychiatric/degenerative disorders, atherosclerosis, coronary artery disease, diabetes mellitus and even carcinogenesis. The present study is designed to determine whether the level of inflammation created by acne vulgaris could be high enough to raise the serum levels of high-sensitive CRP. Materials and Methods: Forty-two patients with moderate and severe acne vulgaris were enrolled, along with 44 age and sex matched healthy blood donors as controls. Hypersensitive-CRP (Hs-CRP was measured in both groups. Results: Hypersensitive-C-reactive protein levels in the case group varied between 0 and 28.1 μg/ml with an average of 2.24 ± 4.87 μg/ml (mean ± standard deviation and a median of 0.6 μg/ml (interquartile range [IQR] =0.3, 1.4 μg/ml. Hs-CRP levels of the control group varied between 0 and 14 μg/ml with an average of 3.12 ± 3.67 μg/ml and a median of 1.5 μg/ml (IQR = 0.55, 5.0 μg/ml. No significant difference of Hs-CRP level between the two groups was seen (t = -0.961, 95% confidence interval: Lower = -2.6942, upper = 0.9377; P = 0.339. Additionally, no significant difference in the level of Hs-CRP was noted between the moderate and severe acne groups (95% confidence interval: Lower = -5.2495, upper = 1.6711; P = 0.165. Conclusion: Acne vulgaris, even in its severe grades (excluding acne fulminans and acne conglobata, does not induce significant inflammation at the systemic level.

  11. Monocyte chemoattractant protein-1 in spinal tuberculosis: -362G/C genetic variant and protein levels in Chinese patients.

    Science.gov (United States)

    Guo, Chaofeng; Zhang, Hongqi; Gao, Qile; He, Dan; Tang, Mingxing; Liu, Shaohua; Deng, Ang; Wang, Yuxiang; Lu, Shijin; Li, Jingsong; Yin, Xinhua; Guo, Qiang

    2014-01-01

    The objective of the study is to explore the possible association of the monocyte chemoattractant protein (MCP)-1-362G/C genetic polymorphism and plasma levels of MCP-1 in patients with spinal tuberculosis (TB). The MCP-1-362G/C (rs2857656) polymorphism and blood levels of MCP-1 in patients with spinal TB and healthy subjects were evaluated and compared. Three hundred thirty-two patients and 336 healthy subjects were genotyped using polymerase chain reaction and Sanger DNA sequencing technology. MCP-1 plasma levels were measured by a solid-phase enzyme-linked immunosorbent assay. When comparisons were made between patients and controls, the frequency of the MCP-1-362*C minor allele (55.4% versus 47.5%, P = 0.004, odds ratio [OR] = 1.376, 95% confidence interval [CI]: 1.109-1.706) and the carriers of the MCP-1-362*C allele (80.7% versus 71.4%, P = 0.005, OR = 1. 657, 95% CI: 1.167-2.403) were over-represented in patients. The mean MCP-1 plasma level in spinal TB patients was significantly higher than in controls (154.44 ± 68.81 pg/mL versus 36.69 ± 21.71 pg/mL, t = -5.85, P < 0.001). The patients with the CC genotype had the highest MCP-1 level (150.63 ± 73.89 pg/mL), followed by those with the GC genotype (108.63 ± 52.09 pg/mL, t = 2.351, P = 0.022) and GG (91.29 ± 54.31 pg/mL, t = 3.091, P = 0.003) homozygotes. We report the association of the -362G/C genetic polymorphism and increased plasma levels of MCP-1 in patients with spinal TB and nominate the -362*C minor allele as a risk factor for spinal TB in the Chinese population.

  12. Crude protein level of pre-starter diets and nutritive solution for broilers

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    Rodrigo Santana Toledo

    2011-10-01

    Full Text Available The objective of the present study was to evaluate the effect of dietary crude protein (CP levels and the use of a nutritive solution via drinking water on the performance of pre-starter broilers. A total of 1,224 male Avian Farm chicks, from one to 40 days of age were used. Birds were distributed in a completely randomized experimental design with a 3 × 2 × 2 factorial arrangement, consisting of three initial body weights (low: 41 g, standard: 45 g and high: 49 g; two crude protein levels at the pre-starter phase (22 and 25% CP,with or without the addition of a nutritive solution, whose nutritional level was similar to the 25% CP pre-starter diet, at a concentration of 5% of the drinking water. Each treatment included six replicates and 17 chicks per experimental unit. At the end of the pre-starter phase, all hens received a diet with 22% CP until day 21 and a diet with 20% CP from the 21st to the 40th day. The use of the pre-starter diet with higher nutritional levels and the nutritive solution enhanced broiler performance. The early nutrient supply via drinking water resulted in better broiler performance and uniformity. However, birds with low initial body weight continued to present lower body weight at market age.

  13. Effect of Dietary Protein Levels on Composition of Odorous Compounds and Bacterial Ecology in Pig Manure

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    Sungback Cho

    2015-09-01

    Full Text Available This study was performed to investigate the effect of different levels of dietary crude protein (CP on composition of odorous compounds and bacterial communities in pig manure. A total of 48 male pigs (average initial body weight 45 kg fed diets containing three levels of dietary CP (20%, 17.5%, and 15% and their slurry samples were collected from the pits under the floor every week for one month. Changes in composition of odorous compounds and bacterial communities were analyzed by gas chromatography and 454 FLX titanium pyrosequencing systems, respectively. Levels of phenols, indoles, short chain fatty acid and branched chain fatty acid were lowest (p<0.05 in CP 15% group among three CP levels. Relative abundance of Bacteroidetes phylum and bacterial genera including Leuconostoc, Bacillus, Atopostipes, Peptonphilus, Ruminococcaceae_uc, Bacteroides, and Pseudomonas was lower (p<0.05 in CP 15% than in CP 20% group. There was a positive correlation (p<0.05 between odorous compounds and bacterial genera: phenol, indole, iso-butyric acid, and iso-valeric acid with Atopostipes, p-cresol and skatole with Bacteroides, acetic acid and butyric acid with AM982595_g of Porphyromonadaceae family, and propionic acid with Tissierella. Taken together, administration of 15% CP showed less production of odorous compounds than 20% CP group and this result might be associated with the changes in bacterial communities especially whose roles in protein metabolism.

  14. Investigation of change of serum immunosuppressive acidic protein levels in gynecological tumors

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Objective: To study the clinical significance for measuring serum immunosuppressive acidic protein(IAP) levels to diagnose and follow up survey patients with the gynecological tumor.Methods: Serum IAP levels were determined by IAP-single radial immunodiffusion test in 235 patients with the gynecological tumor,including 38 cases of benign tumor of ovary,41 cases of malignant tumor of ovary,66 cases of hysteromyoma,34 carcinomas of uterine cervix, 16 endometrial carcinomas,27 cases of chemotherapy,13 cases of recurrence, and the control group was 50 cases health women.Results: Serum IAP level was 889.4±207.8mg/L in malignant ovary tumors,which was significantly higher than that of health women and benign tumors of ovary (P<0.01).In patients with carcinoma of uterine cervix and endometrial carcinoma, their IAP levels were 741.4±212.6mg/L and 763.3±209.4mg/L,which were higher than those of the health women and benign tumor of ovary(P<0.01).After chemotherapy, serum IAP levels of malignant tumor of ovary were decreased;in patients with recurrence of tumor of ovary,IAP levels increased compared with the health women(P<0.01).Incidence of the abnormal value was 100%.Conclusion:Measuring IAP level of the gynecological tumor may be an auxiliary index for monitoring gynecological tumor and identifying benign and malignant tumor.

  15. The Effect of Cinnamon Extract on Serum Proteins Levels of Male Balb/c Mice

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    M Modaresi

    2011-12-01

    Full Text Available Background & Aim: Cinnamon is a plant with many pharmaceutical effects. The present research evaluated the effects of Cinnamon bark extract on serum proteins level in male Balb/c mice. Methods: In this experimental study, 40 small Balb/c mice were chosen and divided into 5 groups: a control group, a case group, and three treatment groups. Normal saline was administered as placebo to the case group while the control group received no injections.. Cinnamon extract in doses of 50, 100 and 200 mg/Kg/48hr were injected intraperitoneally for 20 days to treatment groups. The levels of pre albumin, albumin, alpha-1, alpha-2, beta and gamma globulins were separated electrophoretically and calculated from the pattern of electrophoretogram. Results: The result indicated that the levels of pre albumin decreased significantly in two experimental groups (doses of 50 and 100 mg/kg, the levels of alpha-1 and beta in 200 group, alpha-2 level in 100 and gamma in 100 and 200 groups increased significantly .The injection of 100 mg/Kg/48h extract of Cinnamon decreased (p<0.05 the albumin level in plasma in treatment group as compared to the control group. Conclusion: Results of this study indicated that the serum level of globulins has not changed dramatically by the extract of Cinnamon. Since albumin synthesis occurs in the liver cells‚ thus administration of Cinnamon may affect the function of liver cells.

  16. Mapping Hydrophobicity on the Protein Molecular Surface at Atom-Level Resolution

    Science.gov (United States)

    Nicolau Jr., Dan V.; Paszek, Ewa; Fulga, Florin; Nicolau, Dan V.

    2014-01-01

    A precise representation of the spatial distribution of hydrophobicity, hydrophilicity and charges on the molecular surface of proteins is critical for the understanding of the interaction with small molecules and larger systems. The representation of hydrophobicity is rarely done at atom-level, as this property is generally assigned to residues. A new methodology for the derivation of atomic hydrophobicity from any amino acid-based hydrophobicity scale was used to derive 8 sets of atomic hydrophobicities, one of which was used to generate the molecular surfaces for 35 proteins with convex structures, 5 of which, i.e., lysozyme, ribonuclease, hemoglobin, albumin and IgG, have been analyzed in more detail. Sets of the molecular surfaces of the model proteins have been constructed using spherical probes with increasingly large radii, from 1.4 to 20 Å, followed by the quantification of (i) the surface hydrophobicity; (ii) their respective molecular surface areas, i.e., total, hydrophilic and hydrophobic area; and (iii) their relative densities, i.e., divided by the total molecular area; or specific densities, i.e., divided by property-specific area. Compared with the amino acid-based formalism, the atom-level description reveals molecular surfaces which (i) present an approximately two times more hydrophilic areas; with (ii) less extended, but between 2 to 5 times more intense hydrophilic patches; and (iii) 3 to 20 times more extended hydrophobic areas. The hydrophobic areas are also approximately 2 times more hydrophobicity-intense. This, more pronounced “leopard skin”-like, design of the protein molecular surface has been confirmed by comparing the results for a restricted set of homologous proteins, i.e., hemoglobins diverging by only one residue (Trp37). These results suggest that the representation of hydrophobicity on the protein molecular surfaces at atom-level resolution, coupled with the probing of the molecular surface at different geometric resolutions

  17. Effect of pumpkin seed (Cucurbita pepo) protein isolate on the activity levels of certain plasma enzymes in CCl4-induced liver injury in low-protein fed rats.

    Science.gov (United States)

    Nkosi, C Z; Opoku, A R; Terblanche, S E

    2005-04-01

    The effects of pumpkin seed (Cucurbita pepo) protein isolate on the activity levels of lactate dehydrogenase (LD), alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) against carbon tetrachloride (CCl4)-induced acute liver injury in low-protein fed rats were investigated. A group of male Sprague-Dawley rats maintained on a low-protein diet for 5 days were divided into three subgroups. Two subgroups were injected with carbon tetrachloride and the other group with an equivalent amount of olive oil. Two hours after CCl4 intoxication one of the two subgroups was administered with pumpkin seed protein isolate. All three subgroups of rats were maintained on the low-protein diet for the duration of the investigation. Groups of rats from the different subgroups were killed at 24, 48 and 72 h after their respective treatments. After 5 days on the low-protein diet the activity levels of all four enzymes were significantly higher than their counterparts on a normal balanced diet. CCl4 intoxication resulted in significant increases in the activity levels of all four enzymes investigated. The administration of pumpkin seed protein isolate after CCl4 intoxication resulted in significantly reduced activity levels of all four enzymes. It is concluded that pumpkin seed protein isolate administration was effective in alleviating the detrimental effects associated with protein malnutrition.

  18. Levels of the E2 interacting protein TopBP1 modulate papillomavirus maintenance stage replication

    Energy Technology Data Exchange (ETDEWEB)

    Kanginakudru, Sriramana, E-mail: skangina@iu.edu [Department of Dermatology, Indiana University School of Medicine, Indianapolis, IN (United States); DeSmet, Marsha, E-mail: mdesmet@iupui.edu [Department of Dermatology, Indiana University School of Medicine, Indianapolis, IN (United States); Thomas, Yanique, E-mail: ysthomas@umail.iu.edu [Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN (United States); Morgan, Iain M., E-mail: immorgan@vcu.edu [VCU Philips Institute for Oral Health Research, Virginia Commonwealth University, Richmond, Virginia (United States); Androphy, Elliot J., E-mail: eandro@iu.edu [Department of Dermatology, Indiana University School of Medicine, Indianapolis, IN (United States); Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN (United States)

    2015-04-15

    The evolutionarily conserved DNA topoisomerase II beta-binding protein 1 (TopBP1) functions in DNA replication, DNA damage response, and cell survival. We analyzed the role of TopBP1 in human and bovine papillomavirus genome replication. Consistent with prior reports, TopBP1 co-localized in discrete nuclear foci and was in complex with papillomavirus E2 protein. Similar to E2, TopBP1 is recruited to the region of the viral origin of replication during G1/S and early S phase. TopBP1 knockdown increased, while over-expression decreased transient virus replication, without affecting cell cycle. Similarly, using cell lines harboring HPV-16 or HPV-31 genome, TopBP1 knockdown increased while over-expression reduced viral copy number relative to genomic DNA. We propose a model in which TopBP1 serves dual roles in viral replication: it is essential for initiation of replication yet it restricts viral copy number. - Highlights: • Protein interaction study confirmed In-situ interaction between TopBP1 and E2. • TopBP1 present at papillomavirus ori in G1/S and early S phase of cell cycle. • TopBP1 knockdown increased, over-expression reduced virus replication. • TopBP1 protein level change did not influence cell survival or cell cycle. • TopBP1 displaced from papillomavirus ori after initiation of replication.

  19. Lung surfactant levels are regulated by Ig-Hepta/GPR116 by monitoring surfactant protein D.

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    Taku Fukuzawa

    Full Text Available Lung surfactant is a complex mixture of lipids and proteins, which is secreted from the alveolar type II epithelial cell and coats the surface of alveoli as a thin layer. It plays a crucial role in the prevention of alveolar collapse through its ability to reduce surface tension. Under normal conditions, surfactant homeostasis is maintained by balancing its release and the uptake by the type II cell for recycling and the internalization by alveolar macrophages for degradation. Little is known about how the surfactant pool is monitored and regulated. Here we show, by an analysis of gene-targeted mice exhibiting massive accumulation of surfactant, that Ig-Hepta/GPR116, an orphan receptor, is expressed on the type II cell and sensing the amount of surfactant by monitoring one of its protein components, surfactant protein D, and its deletion results in a pulmonary alveolar proteinosis and emphysema-like pathology. By a coexpression experiment with Sp-D and the extracellular region of Ig-Hepta/GPR116 followed by immunoprecipitation, we identified Sp-D as the ligand of Ig-Hepta/GPR116. Analyses of surfactant metabolism in Ig-Hepta(+/+ and Ig-Hepta(-/- mice by using radioactive tracers indicated that the Ig-Hepta/GPR116 signaling system exerts attenuating effects on (i balanced synthesis of surfactant lipids and proteins and (ii surfactant secretion, and (iii a stimulating effect on recycling (uptake in response to elevated levels of Sp-D in alveolar space.

  20. Lung surfactant levels are regulated by Ig-Hepta/GPR116 by monitoring surfactant protein D.

    Science.gov (United States)

    Fukuzawa, Taku; Ishida, Junji; Kato, Akira; Ichinose, Taro; Ariestanti, Donna Maretta; Takahashi, Tomoya; Ito, Kunitoshi; Abe, Jumpei; Suzuki, Tomohiro; Wakana, Shigeharu; Fukamizu, Akiyoshi; Nakamura, Nobuhiro; Hirose, Shigehisa

    2013-01-01

    Lung surfactant is a complex mixture of lipids and proteins, which is secreted from the alveolar type II epithelial cell and coats the surface of alveoli as a thin layer. It plays a crucial role in the prevention of alveolar collapse through its ability to reduce surface tension. Under normal conditions, surfactant homeostasis is maintained by balancing its release and the uptake by the type II cell for recycling and the internalization by alveolar macrophages for degradation. Little is known about how the surfactant pool is monitored and regulated. Here we show, by an analysis of gene-targeted mice exhibiting massive accumulation of surfactant, that Ig-Hepta/GPR116, an orphan receptor, is expressed on the type II cell and sensing the amount of surfactant by monitoring one of its protein components, surfactant protein D, and its deletion results in a pulmonary alveolar proteinosis and emphysema-like pathology. By a coexpression experiment with Sp-D and the extracellular region of Ig-Hepta/GPR116 followed by immunoprecipitation, we identified Sp-D as the ligand of Ig-Hepta/GPR116. Analyses of surfactant metabolism in Ig-Hepta(+/+) and Ig-Hepta(-/-) mice by using radioactive tracers indicated that the Ig-Hepta/GPR116 signaling system exerts attenuating effects on (i) balanced synthesis of surfactant lipids and proteins and (ii) surfactant secretion, and (iii) a stimulating effect on recycling (uptake) in response to elevated levels of Sp-D in alveolar space.

  1. Intermediate divergence levels maximize the strength of structure-sequence correlations in enzymes and viral proteins.

    Science.gov (United States)

    Jackson, Eleisha L; Shahmoradi, Amir; Spielman, Stephanie J; Jack, Benjamin R; Wilke, Claus O

    2016-07-01

    Structural properties such as solvent accessibility and contact number predict site-specific sequence variability in many proteins. However, the strength and significance of these structure-sequence relationships vary widely among different proteins, with absolute correlation strengths ranging from 0 to 0.8. In particular, two recent works have made contradictory observations. Yeh et al. (Mol. Biol. Evol. 31:135-139, 2014) found that both relative solvent accessibility (RSA) and weighted contact number (WCN) are good predictors of sitewise evolutionary rate in enzymes, with WCN clearly out-performing RSA. Shahmoradi et al. (J. Mol. Evol. 79:130-142, 2014) considered these same predictors (as well as others) in viral proteins and found much weaker correlations and no clear advantage of WCN over RSA. Because these two studies had substantial methodological differences, however, a direct comparison of their results is not possible. Here, we reanalyze the datasets of the two studies with one uniform analysis pipeline, and we find that many apparent discrepancies between the two analyses can be attributed to the extent of sequence divergence in individual alignments. Specifically, the alignments of the enzyme dataset are much more diverged than those of the virus dataset, and proteins with higher divergence exhibit, on average, stronger structure-sequence correlations. However, the highest structure-sequence correlations are observed at intermediate divergence levels, where both highly conserved and highly variable sites are present in the same alignment.

  2. Plasma lactate, GH and GH-binding protein levels in exercise following BCAA supplementation in athletes.

    Science.gov (United States)

    De Palo, E F; Gatti, R; Cappellin, E; Schiraldi, C; De Palo, C B; Spinella, P

    2001-01-01

    Branched chain amino acids (BCAA) stimulate protein synthesis, and growth hormone (GH) is a mediator in this process. A pre-exercise BCAA ingestion increases muscle BCAA uptake and use. Therefore after one month of chronic BCAA treatment (0.2 gkg(-1) of body weight), the effects of a pre-exercise oral supplementation of BCAA (9.64 g) on the plasma lactate (La) were examined in triathletes, before and after 60 min of physical exercise (75% of VO2 max). The plasma levels of GH (pGH) and of growth hormone binding protein (pGHBP) were also studied. The end-exercise La of each athlete was higher than basal. Furthermore, after the chronic BCAA treatment, these end-exercise levels were lower than before this treatment (8.6+/-0.8 mmol L(-1) after vs 12.8+/-1.0 mmol L(-1) before treatment; p BCAA chronic treatment, this end-exercise pGHBP was 738+/-85 pmol L(-1) before vs 1691+/-555 pmol L(-1) after. pGH/pGHBP ratio was unchanged in each athlete and between the groups, but a tendency to increase was observed at end-exercise. The lower La at the end of an intense muscular exercise may reflect an improvement of BCAA use, due to the BCAA chronic treatment. The chronic BCAA effects on pGH and pGHBP might suggest an improvement of muscle activity through protein synthesis.

  3. Renal and urinary levels of endothelial protein C receptor correlate with acute renal allograft rejection.

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    Lionel Lattenist

    Full Text Available The Endothelial Protein C Receptor (EPCR is expressed on leukocytes, on endothelium of large blood vessels and to a lesser extent on capillaries. Membrane bound EPCR plays an important role in the activation of protein C which has anticoagulant, anti-inflammatory and cytoprotective effects. After cleavage by a protease EPCR is also found as a soluble protein. Acute rejection of kidney allografts can be divided in T-cell-mediated rejection (TCMR and antibody-mediated (ABMR rejection. The latter is characterized by strong activation of coagulation. Currently no reliable non-invasive biomarkers are available to monitor rejection. Renal biopsies were available from 81 renal transplant patients (33 without rejection, 26 TCMR and 22 ABMR, we had access to mRNA material, matched plasma and urine samples for a portion of this cohort. Renal EPCR expression was assessed by RT-PCR and immunostaining. Plasma and urine sEPCR levels were measured by ELISA. ABMR patients showed higher levels of EPCR mRNA than TCMR patients. EPCR expression on glomeruli was significantly elevated in ABMR patients than in TCMR or control patients. In the peritubular capillaries EPCR expression was higher in ABMR patients than in control patients. EPCR expression was higher in tubules and arteries of rejection patients than in control patients. Plasma sEPCR levels did not differ. Urine sEPCR levels were more elevated in the ABMR group than in patients with TCMR or without rejection. ROC analysis demonstrated that urinary sEPCR is appropriate to discriminate between ABMR patients and TCMR or control patients. We conclude that urinary sEPCR could be a novel non-invasive biomarker of antibody mediated rejection in renal transplantation.

  4. Comparison Of Serum Calcium, Phosphorus And Total Protein Levels, In Pregnancy With Or Without Hypertensive Disorders

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    Rashid Pooraei M

    2005-06-01

    Full Text Available Background: Pre-eclampsia is characterized by hypertension development and proteinuria during pregnancy. Hypertension disorder is a leading cause of maternal and fetal morbidity and death in worldwide. Although the pathophysiology of hypertension during pregnancy is unclear, but there is consensus that early diagnosis and aggressive treatment is warranted to prevent complicated to both fetus and mother. The changes of serum trace elements during pregnancy are paramount important to predict and good understanding the situation of patients. The aim of this study was about this issue. Materials and Methods: In a case-control study we investigated the possible differences in the level of serum calcium, phosphorus and total protein in 55 healthy pregnant and 52 pregnant with hypertensive disease at 32-40 weeks of gestational age during the recent two years in Loghman hospital of Tehran. Some information such as age, blood group, parity number and blood pressure was taken from patients by a questionnaire. Results: The case population consisted of 22 sever preeclampsia, 15 mild preeclampsia, 8 eclampsia, and 7 chronic hypertensive. The mean serum calcium concentration (mg/dl was 9.180.74 in control group, 8.810.9 in mild preeclampsia, 7.850.38 in sever preeclampsia, 7.83 0.47 in eclampsia, 8.91 0.3 in chronic blood pressure. The mean serum phosphorus (mg/dl level observed, 4.27 in sever preeclampsia, 3.74 in eclampsia, 3.59 in mild eclampsia, 4.09 in chronic blood pressure, and 3.43 in control pregnant women. The mean serum total protein concentration level in sever preeclampsia and eclampsia was 5.46 and 5.04 mg/dl respectively. Conclusion: In conclusion, sever preeclampsia and eclampsia are associated with decreased level of calcium, total protein, and increased concentration of phosphorus.

  5. Brain and hepatic Hsp70 protein levels in heat-acclimated broiler chickens during heat stress

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    EN Guerreiro

    2004-12-01

    Full Text Available In the present study we have investigated the effects of heat acclimation on brain and hepatic Hsp70 protein levels and body temperature of broiler chickens in response to gradual heat stress. Two groups of broilers were raised up to 47 days of age under distinct temperature conditions: thermoneutral (TN, according to bird age or hot environmental (HS, 31-33°C. At 46 days of age, the birds reared at high ambient temperature were transferred to thermoneutrality conditions. After 18 h, these birds and the birds reared at thermoneutral temperature were submitted to gradual heat stress in a climatic chamber so that environment temperature was increased from 28 to 40ºC at a rate of 2ºC/h. Colonic temperature was measured using a thermometer sensor probe at each two hours, and hepatic and brain tissues were collected immediately after slaughter in order to assess Hsp70 protein level by Western blotting analysis. The colonic temperatures of birds reared at high temperature increased steeply during the first 2 h of heat stress (1.06ºC/h and more slowly thereafter (0.59ºC/h. Broilers reared at thermoneutral temperature showed a small increase in the first 4 h of heat stress (0.18ºC/h and then colonic temperature increased sharply (0.72ºC/h. Nevertheless, both groups presented similar final colonic temperature by the end of the stress period. Hsp70 levels (ng Hsp70 µg total protein-1 did not change in the liver or brain of the birds reared at high temperature. On the other hand, both liver and brain Hsp70 levels increased significantly during heat stress in the animals reared at thermoneutrality, with a higher expression of this peptide in brain tissue.

  6. RELATIONSHIP AMONG COX-2 PROTEIN EXPRESSION, PGs LEVELS AND BIOLOGIC BEHAVIOR IN OVARIAN CARCINOMA TISSUES

    Institute of Scientific and Technical Information of China (English)

    王敏; 王欣彦; 唐丽霞; 高岩

    2004-01-01

    Objective: To study the relationship among cyclooxygenase-2 (COX-2) protein expression, prostaglandins levels and biologic behavior in ovarian carcinoma tissues. Methods: The expression of COX-2 protein, levels of prostaglandin (PG)E2, 6-keto-PGF1( and thromboxane (TX)B2 in 54 biopsy specimens from patients with ovarian serous tumors which included three groups: 33 samples of ovarian serous carcinoma; 10 samples of borderline ovarian serous tumors and 11 samples of benign ovarian serous tumors and 10 samples of normal ovarian tissues were detected by Western blot analysis and radioimmunoassay to investigate their clinical significance. Results: The expression of COX-2 protein (82%, 27/33) and its relative content (20.08±3.53) in ovarian serous carcinoma tissues were statistically higher than those in benign ovarian serous tumor tissues and normal ovary tissues i.e., 0 and (15.04(0.12), 0 and (15.33(0.60) (P0.05). The levels of PGE2, 6-keto-PGF1( and TXB2 showed no significant differences in ovarian carcinoma tissues with different clinical stages (I to II and III to IV), different histological grades, with or without ascites and lymph metastasis. COX-2 expression was correlated with the levels of PGE2, 6-KETO-PGF1( and TXB2 (P<0.01). Conclusion: Our data suggest that COX-2 overexpression leads to increased PGE2, 6-KETA-PGF1( and TXB2 biosynthesis, which may be mechanisms underlying the contribution of COX-2 to the development of ovarian serous carcinoma. BGF2, 6-keto-PGF1( and TXB2 may be helpful parameters of diagnosis and differentiate diagnosis in ovarian serous carcinoma.

  7. Serum Levels of Surfactant Proteins in Patients with Combined Pulmonary Fibrosis and Emphysema (CPFE.

    Directory of Open Access Journals (Sweden)

    Andriana I Papaioannou

    Full Text Available Emphysema and idiopathic pulmonary fibrosis (IPF present either per se or coexist in combined pulmonary fibrosis and emphysema (CPFE. Serum surfactant proteins (SPs A, B, C and D levels may reflect lung damage. We evaluated serum SP levels in healthy controls, emphysema, IPF, and CPFE patients and their associations to disease severity and survival.122 consecutive patients (31 emphysema, 62 IPF, and 29 CPFE and 25 healthy controls underwent PFTs, ABG-measurements, 6MWT and chest HRCT. Serum levels of SPs were measured. Patients were followed-up for 1-year.SP-A and SP-D levels differed between groups (p = 0.006 and p<0.001 respectively. In post-hoc analysis, SP-A levels differed only between controls and CPFE (p<0.05 and CPFE and emphysema (p<0.05. SP-D differed between controls and IPF or CPFE (p<0.001 for both comparisons. In IPF SP-B correlated to pulmonary function while SP-A, correlated to the Composite Physiological Index (CPI. Controls current smokers had higher SP-A and SP-D levels compared to non-smokers (p = 0.026 and p = 0.023 respectively. SP-D levels were higher in CPFE patients with extended emphysema (p = 0.042. In patients with IPF, SP-B levels at the upper quartile of its range (≥26 ng/mL presented a weak association with reduced survival (p = 0.05.In conclusion, serum SP-A and SP-D levels were higher where fibrosis exists or coexists and related to disease severity, suggesting that serum SPs relate to alveolar damage in fibrotic lungs and may reflect either local overproduction or overleakage. The weak association between high levels of SP-B and survival needs further validation in clinical trials.

  8. Carvacrol attenuates serum levels of total protein, phospholipase A2 and histamine in asthmatic guinea pig

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    Mohammad Hossein Boskabady

    2016-11-01

    Full Text Available Objective: Pharmacological effects of carvacrol such as its anti-inflammatory activities have been shows. In this study the effects of carvacrol on serum levels of total protein (TP, phospholipase A2 (PLA2 and histamine in sensitized guinea pigs was evaluated. Materials and Methods: Sensitized guinea pigs were given drinking water alone (group S, drinking water containing three concentrations of carvacrol (40, 80 and 160 µg/ml or dexamethasone. Serum levels of TP, PLA2 and histamine were examined I all sensitized groups as well as a non-sensitized control group (n=6 for each group. Results: In sensitized animals, serum levels of TP, PLA2 and histamine were significantly increased compared to control animals (p

  9. IL-18 and IL-18 binding protein levels in patients with dengue virus infection

    Institute of Scientific and Technical Information of China (English)

    Chintana Chirathaworn; Yong Poovorawan; Viboonsak Vuthitanachot

    2010-01-01

    Objective:To determine the levels ofIL-18 andIL-18binding protein(BP) in patients with dengue virus infection.Methods: Acute and convalescent sera were collected from each patient. Control group was sera from blood donors. The levels of both IL-18 andIL-18BP were measured byELISA assays.Results: It was shown thatIL-18 andIL-18BPlevels were significantly higher in patients when compared with controls. In addition, the level ofIL-18BP was lower in convalescent than in acute sera.Conclusions: These data suggest that bothIL-18 andIL-18BP production was induced following dengue virus infection. Investigating the regulation of IL-18 by its natural regulator could lead to further understanding of the immune response or immunopathogenesis following dengue virus infection.

  10. C-reactive protein level as a predictor of mortality in liver disease patients with bacteremia

    DEFF Research Database (Denmark)

    Janum, Sine H; Søvsø, Morten; Gradel, Kim O

    2011-01-01

    Abstract Background and objective. C-reactive protein (CRP) is synthesized in the liver in response to inflammation, and CRP is a widely used marker of sepsis. In bacteremia the initial CRP level is an independent predictor of mortality. Since the CRP response in patients with chronic liver disease...... is lower than in patients without liver disease the objective was to assess whether CRP levels in chronic liver disease and bacteremia was associated with case fatality. Patients. The study enrolled 105 patients with chronic liver disease and bacteremia as well as 202 patients with bacteremia...... was observed. Conclusion. Mortality associated with bacteremia is increased in patients with chronic liver disease and it is correlated with Child-Turcotte-Pugh score. The prognostic information of initial CRP levels in patients with chronic liver disease is weak. The clinical management of patients...

  11. Increased plasma amyloid beta protein 1-42 levels in Down syndrome.

    Science.gov (United States)

    Mehta, P D; Dalton, A J; Mehta, S P; Kim, K S; Sersen, E A; Wisniewski, H M

    1998-01-23

    Amyloid beta protein 1-40 (A beta40) and A beta42 levels were quantitated in plasma from 43 persons with Down syndrome (DS; 26-68 years of age), 43 age-matched normal controls, and 19 non-DS mentally retarded (MR) persons (26-91 years of age) by using a sandwich enzyme linked immunosorbent assay. A beta40 levels were higher in DS and MR than controls, but were similar between DS and MR groups. A beta42 levels were higher in DS than controls or MR persons. The ratios of A beta42/A beta40 were higher in DS than controls or MR persons. The findings are consistent with those seen in DS brains.

  12. Study of p53 protein expression levels from irradiated peripheral blood lymphocytes for biodosimetry

    Energy Technology Data Exchange (ETDEWEB)

    Cavalcanti, M.B.; Fernandes, T.S. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Energia Nuclear; Amaral, A. [Universite Paris XII (UPXII) (France); Melo, J.A. [Centro de Radioterapia de Pernambuco (CERAPE), PE (Brazil); Neves, M.A.B.; Machado, C.G.F, E-mail: maribrayner@yahoo.com.br [Fundacao de Hematologia e Hemoterapia de Pernambuco, PE (Brazil)

    2005-07-01

    Biodosimetry can be defined as the investigation of radioinduced biological effects in order to correlate them with the absorbed dose. Scoring of unstable chromosomal aberrations and micronuclei, from in vitro irradiated peripheral blood lymphocytes, is commonly used for biodosimetry based on cytogenetic analysis. However, this method of analysis is time-consuming, which may represent a pitfall when fast investigation of a possible exposure to ionizing radiation (IR) is needed. The interaction of IR with the living cell can cause injuries in the DNA molecules. However, normal cells possess mechanisms of repair that are capable to correct those damages. During the repair process of the DNA various proteins are expressed. Among these proteins, p53 plays an important role. This protein is a transcription factor that helps in the maintenance of the genomic integrity. p53 protein is found into the cytoplasm in reduced concentrations and has a short average life. However, expression of p53 protein can be induced by DNA harmful radioinduced, which increases the concentration and the average life of this protein, making possible its detection. Thus, the correlation between the increasing of p53 expression and the irradiation may constitute a fast and reliable method of individual monitoring in cases of accidental or suspected exposures to IR. In this context, the objective of this research was to evaluate the p53 protein expression levels from lymphocytes of the human peripheral blood after in vitro irradiation. For this, samples of peripheral blood from healthy individuals were irradiated with known doses. Lymphocytes were separated on ficoll gradient by centrifugation and re-suspended at 1x 10{sub 6}/mL in RPMI medium enriched with fetal calf serum. Hence, lymphocytes were incubated in 5% CO{sub 2} at 37 deg C prior to the methodology of flow cytometry, using intranuclear antigens for the quantification of p53. In this report, the methodology performed and the results

  13. Effect of dietary protein level on ewe milk yield and on air quality under different ventilation rates

    Directory of Open Access Journals (Sweden)

    A. Sevi

    2010-01-01

    Full Text Available The efficiency of dietary N utilization for milk protein synthesis in dairy animals is quite low (15 to 35% (NRC, 1988; Tamminga, 1992, therefore farmers are driven to use high protein level diets for sustaining milk production in lactating animals. Previous experiments have demonstrated that an increase in the protein level of diet from 13 to 16% resulted in higher blood urea concentrations (Jaime and Purroy, 1995 and increased N excretion in urine in sheep (Gonzalez et al., 1984.

  14. Reduced NM23 Protein Level Correlates With Worse Clinicopathologic Features in Colorectal Cancers

    Science.gov (United States)

    Yang, Tian; Chen, Bo-Zan; Li, Dan-Feng; Wang, Huai-Ming; Lin, Xiao-Sheng; Wei, Hong-Fa; Zeng, Yong-Ming

    2016-01-01

    Abstract The clinical value of a prominent metastasis suppressor, nonmetastatic protein 23 (NM23), remains controversial. In this study, we examined the correlation between NM23 protein levels and the clinicopathologic features of colorectal cancers (CRC), and assessed the overall prognostic value of NM23 for CRC. Embase, PubMed, Web of Science, and other scientific literature databases were exhaustively searched to identify relevant studies published prior to June 31, 2015. The methodological qualities of selected studies were scored based on the critical appraisal skills program (CASP) criteria, as independently assessed by 2 reviewers. NM23 protein levels in tumor tissues of CRC patients were examined in relation to Dukes stage, differentiation grade, T-stage, lymph node metastasis status, and overall survival (OS). STATA software version 12.0 (Stata Corp, College Station, TX) was used for statistical analysis of data pooled from selected studies. Nineteen cohort studies met the inclusion criteria for present study and contained a combined total of 2148 study subjects. Pooled odd ratios (ORs) for NM23 expression revealed that reduced NM23 protein levels in CRC tumor tissues correlated with Dukes stage C and D (OR = 1.89, 95% CI: 1.06–3.39, P = 0.032), poor differentiation grades (OR = 1.41, 95% CI: 1.03–1.94, P = 0.032), and positive lymph node metastasis status (OR = 3.21, 95% CI: 1.95–5.29, P < 0.001). On the other hand, no such correlations were evident with T-stage T3-4 (OR = 1.56, 95% CI: 0.60–4.06, P = 0.367) or OS (OR = 0.79, 95% CI: 0.58–1.08, P = 0.138). Our analysis of pooled data found that NM23 expression is reduced in CRC tissues and low NM23 levels tightly correlate with higher Dukes stages, poorer differentiation grade, and positive lymph node metastases. However, NM23 levels did not influence the OS in CRC patients. PMID:26825905

  15. Blood parameters in growing pigs fed increasing levels of bacterial protein meal

    DEFF Research Database (Denmark)

    Hellwing, Anne Louise Frydendahl; Tauson, Anne-Helene; Skrede, Anders

    2007-01-01

    The experiment investigated the effects of increasing dietary levels of bacterial protein meal (BPM) on various blood parameters reflecting protein and fat metabolism, liver function, and purine base metabolism in growing pigs. Sixteen barrows were allocated to four different experimental diets......, 45 kg, and 77 kg. The blood parameters reflecting fat metabolism and liver funtion were not affected by diet. Both the plasma albumin and uric acid concentrations tended to decrease (P = 0.07 and 0.01, respectively) with increasing dietary BPM content, whereas the plasma glucose concentration tended...... to increase (P = 0.07) with increasing dietary BPM content. It was concluded that up to 50% of the nitrogen could be derived from BPM without affecting metabolic function, as reflected in the measured blood parameters....

  16. Data in support of DPF2 regulates OCT4 protein level and nuclear distribution

    Directory of Open Access Journals (Sweden)

    Chao Liu

    2015-12-01

    Full Text Available DPF2, also named ubi-d4/requiem (REQU, interacts with a protein complex containing OCT4. This paper provides data in support of the research article entitled “DPF2 regulates OCT4 protein level and nuclear distribution”. The highlights include: (1 Denature-immunoprecipitation assay revealed ubiquitination of OCT4 in pluripotent H9 cells, which was enhancedby MG132, a proteasome inhibitor. (2 Well colocalization of ectopic OCT4 and FLAG-Ub was found in HeLa cells, which was also increased by MG132. (3 MG132 treatment decreased DPF2 cytoplasmic expression in vivo. These data give insights into how proteasome inhibition contributes to studying ubiquitnation of OCT4.

  17. Utility of C-Reactive Protein Levels for Early Prediction of Dengue Severity in Adults

    Directory of Open Access Journals (Sweden)

    Chien-Chih Chen

    2015-01-01

    Full Text Available Dengue has broad clinical presentation with unpredictable clinical evolution and outcome. We aimed to evaluate the utility of C-reactive protein (CRP levels for distinguishing between mild and severe cases in the early phase of the dengue illness. We retrospectively evaluated adults with dengue from 2006 to 2014, according to 1997 and 2009 World Health Organization (WHO criteria for severity. Of 191 included patients, 32.9% had nonshock dengue hemorrhagic fever (DHF, 3.1% dengue shock syndrome (DSS, and 7.9% severe dengue. The risk of DHF/DSS and severe dengue is significantly related to the increasing levels of CRP. Of 191 patients, 97 had CRP levels measured during the febrile (days 1–3; 85 during the critical (days 4–6; and 9 during the convalescent (days 7–10 illness phases. During the febrile phase, there was significant higher CRP level for DSS versus DF/nonshock DHF and severe dengue versus nonsevere dengue, with CRP cutoff level 30.1 mg/L (area under the receiver operating characteristic curve (AUC, 0.938; 100% sensitivity, 76.3% specificity and 24.2 mg/L (AUC, 0.717; 70% sensitivity, 71.3% specificity, respectively. Our study highlights the utility of the CRP levels in early prediction of DSS and severe dengue in adult patients.

  18. C-reactive protein levels are influenced by common IL-1 gene variations.

    Science.gov (United States)

    Berger, Peter; McConnell, Joseph P; Nunn, Martha; Kornman, Kenneth S; Sorrell, Julian; Stephenson, Katherine; Duff, Gordon W

    2002-02-21

    Elevated markers of systemic inflammation are associated with the development of acute coronary syndromes, but there is no current explanation for increased inflammation in overtly healthy individuals. The influence of genetic control of the inflammatory response on the observed variability is unknown. We studied the frequency of four polymorphisms in interleukin (IL) 1 genes, known to modulate inflammation, in 454 individuals undergoing coronary angiography and analysed their influence on plasma C-reactive protein (CRP) and fibrinogen levels. Females and smokers had higher levels of CRP than males (Pi = 0.001) and non-smokers (Pi = 0.001). Patients with genotype 2.2 for the IL-1B(+3954) polymorphism had twice the median CRP levels of patients who were genotype 1.1 (4.33 vs 2.01 mg/l; P = 0.001). Patients with genotype 1.2 or 2.2 at the IL-1A(+4845) polymorphism also had higher median CRP (2.92 vs 2.05 mg/l, Pi = 0.023). In multivariate analyses, CRP levels remained significantly associated with IL-1 polymorphisms after adjustment for smoking, gender and age. Fibrinogen levels had similar associations with the IL-1 genotypes. These data indicate that IL-1 gene polymorphisms known to affect the inflammatory response are highly related to plasma levels of CRP and fibrinogen in patients referred for coronary angiography.

  19. Efficacy of erythrocyte sedimentation rate and C-reactive protein level in determining periprosthetic hip infections.

    Science.gov (United States)

    Costa, Christopher R; Johnson, Aaron J; Naziri, Qais; Maralunda, German A; Delanois, Ronald E; Mont, Michael A

    2012-04-01

    The diagnosis of periprosthetic hip infections is often challenging. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) level blood laboratory tests are commonly used to aid in the diagnosis. We studied the sensitivity, specificity, and false-negative rates of ESR and CRP level in a prospective group of patients who underwent revision total hip arthroplasty between 2000 and 2008. Seventy-seven patients with periprosthetic hip infections and ESR and CRP data were identified. Chi-square analysis was performed to determine the significance of false-negatives, compared with sex, body mass index, primary diagnosis, infection type, and immunity status. ESR had 89% sensitivity and 69% specificity. CRP level had 93% sensitivity and 40% specificity. The false-negative rate was 10.8% for ESR and 7% for CRP level. The false-negative rate for ESR and CRP level combined (with either result positive) was 3%. All false-negatives in the combined group were immunocompromised. Chi-square analysis did not find a significant correlation between false-negatives and any other variables. ESR and CRP level are useful in the diagnosis of periprosthetic hip infections. Ordering these tests concurrently reduces the chance of false-negative results.

  20. Serum Growth Arrest Specific Protein 6 (Gas-6 Levels in Patients with Schizophrenia

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    Fethullah Gerin

    2016-03-01

    Full Text Available Objective: We have investigated serum growth ar­rest-specific protein 6 (GAS-6 levels from patients with schizophrenia divided into acute phase remission phases as well as control group. Methods: This study was conducted in Psychiatry De­partment of Istanbul University Cerrahpasa Medical Fac­ulty. The patients who were diagnosed with schizophrenia after regular psychiatric examination according to DSM-IV criteria (n=22 as well as control subjects were includ­ed in the study. Schizophrenia patients with acute phase and remission phase were evaluated by Positive and Negative Syndrome Scale (PANSS and Clinical global Impression Scale (CGI-S. The serum GAS-6 levels of schizophrenia patients during acute phase and remission phase were compared with the serum GAS-6 levels of healthy controls. Serum GAS-6 levels were measured by commercial ELISA kits. Results: No difference was found in serum GAS-6 levels among the three groups; schizophrenia with acute phase, schizophrenia with remission phase, and controls. There were no correlations between serum GAS-6 levels and PANSS and CGI scores. Conclusion: To reach a definitive data and better in­terpretation about the relationship between GAS-6 and schizophrenia, future studies with larger groups of pa­tients with schizophrenia subdivided by drug naïve and treated with antipsychotics/other treatment modalities and controls are needed.

  1. Mechanical spectroscopy of retina explants at the protein level employing nanostructured scaffolds.

    Science.gov (United States)

    Mayazur Rahman, S; Reichenbach, Andreas; Zink, Mareike; Mayr, Stefan G

    2016-04-14

    Development of neuronal tissue, such as folding of the brain, and formation of the fovea centralis in the human retina are intimately connected with the mechanical properties of the underlying cells and the extracellular matrix. In particular for neuronal tissue as complex as the vertebrate retina, mechanical properties are still a matter of debate due to their relation to numerous diseases as well as surgery, where the tension of the retina can result in tissue detachment during cutting. However, measuring the elasticity of adult retina wholemounts is difficult and until now only the mechanical properties at the surface have been characterized with micrometer resolution. Many processes, however, such as pathological changes prone to cause tissue rupture and detachment, respectively, are reflected in variations of retina elasticity at smaller length scales at the protein level. In the present work we demonstrate that freely oscillating cantilevers composed of nanostructured TiO2 scaffolds can be employed to study the frequency-dependent mechanical response of adult mammalian retina explants at the nanoscale. Constituting highly versatile scaffolds with strong tissue attachment for long-term organotypic culture atop, these scaffolds perform damped vibrations as fingerprints of the mechanical tissue properties that are derived using finite element calculations. Since the tissue adheres to the nanostructures via constitutive proteins on the photoreceptor side of the retina, the latter are stretched and compressed during vibration of the underlying scaffold. Probing mechanical response of individual proteins within the tissue, the proposed mechanical spectroscopy approach opens the way for studying tissue mechanics, diseases and the effect of drugs at the protein level.

  2. Evaluation of behaviour in stabled draught horse foals fed diets with two protein levels.

    Science.gov (United States)

    Sartori, C; Guzzo, N; Normando, S; Bailoni, L; Mantovani, R

    2017-01-01

    The present work is aimed at evaluating the behaviour of Italian Heavy Draught Horse (IHDH) foals reared in semi-covered stables and fed two isoenergetic total mixed rations with different dietary protein levels (13.2% and 10.6% of CP on dry matter). The study was prompted by the restrictions for nitrate emissions in farms of the European Nitrate Directive. One suggested solution is to reduce dietary protein while maintaining normal performance and welfare, but there is a lack of literature in studies of horses. The behaviours of 20 foals of 437±60 kg of BW, aged 379±37 days and stabled in four pens by sex (S) and diet (D) were video recorded and analysed to build a suitable ethogram including 18 behaviours in six categories: ingestion, resting, maintenance, movement, social activities, other. The percentage of the daily time spent in each behavioural category and single behaviours was analysed via a single traits GLM including S, D and their interaction. Daily activity was consistent with existing literature: foals spent about 33% of the day in ingestion activities and 41% in resting, whereas social interactions constituted 8% of the time and individual maintenance <2%. Concerning diet, foals fed high protein spent more time in movement (19.62±0.73% of day v. 10.45±0.73% in low-protein (LP) foals; P⩽0.001), whereas the LP group increased resting (43.42±1.12% v. 38.02±1.12%; P⩽0.001). No stereotypies were found, and daily activity followed the typical values for draught breeds for foals in both dietary groups, a result that suggests the maintenance of well-being after dietary protein reduction. This result, together with the findings of a companion study showing no changes in growth performances of foals, showed that a reduction of CP in foal diet is reconcilable with the maintenance of performance and welfare.

  3. Single prolonged stress enhances hippocampal glucocorticoid receptor and phosphorylated protein kinase B levels

    Science.gov (United States)

    Eagle, Andrew L.; Knox, Dayan; Roberts, Megan M.; Mulo, Kostika; Liberzon, Israel; Galloway, Matthew P.; Perrine, Shane A.

    2012-01-01

    Animal models of posttraumatic stress disorder (PTSD) can explore neurobiological mechanisms by which trauma enhances fear and anxiety reactivity. Single prolonged stress (SPS) shows good validity in producing PTSD-like behavior. While SPS-induced behaviors have been linked to enhanced glucocorticoid receptor (GR) expression, the molecular ramifications of enhanced GR expression have yet to be identified. Phosphorylated protein kinase B (pAkt) is critical for stress-mediated enhancement in general anxiety and memory, and may be regulated by GRs. However, it is currently unknown if pAkt levels are modulated by SPS, as well as if the specificity of GR and pAkt related changes contribute to anxiety-like behavior after SPS. The current study set out to examine the effects of SPS on GR and pAkt protein levels in the amygdala and hippocampus and to examine the specificity of these changes to unconditioned anxiety-like behavior. Levels of GR and pAkt were increased in the hippocampus, but not amygdala. Furthermore, SPS had no effect on unconditioned anxiety-like behavior suggesting that generalized anxiety is not consistently observed following SPS. The results suggest that SPS-enhanced GR expression is associated with phosphorylation of Akt, and also suggest that these changes are not related to an anxiogenic phenotype. PMID:23201176

  4. Cold induced changes in lipid, protein and carbohydrate levels in the tropical insect Gromphadorhina coquereliana.

    Science.gov (United States)

    Chowanski, Szymon; Lubawy, Jan; Spochacz, Marta; Ewelina, Paluch; Grzegorz, Smykalla; Rosinski, Grzegorz; Slocinska, Malgorzata

    2015-05-01

    Insects cope with thermal stressors using mechanisms such as rapid cold hardening and acclimation. These mechanisms have been studied in temperate insects, but little is known about their use by tropical insects in response to cold stress. Here, we investigated whether cold stress (1×8 h and 3×8 h at 4°C) triggers a metabolic response in the Madagascar cockroach Gromphadorhina coquereliana. We examined the effects of cold on the levels of selected metabolites in the fat body tissue of G. coquereliana. After cold exposure, we found that the quantity of total protein increased significantly in the insect fat body, whereas glycogen decreased slightly. Using antibodies, we observed upregulation of AQP-like proteins and changes in the HSP70 levels in the fat body of G. coquereliana when exposed to cold. We also examined the content and nature of the free sugars in the G. coquereliana hemolymph and discovered an increase in the levels of polyols and glucose in response to cold stress. These results suggest an important role of the fat body tissue of tropical insects upon cold exposure. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Effect of Dietary Protein Levels on Composition of Odorous Compounds and Bacterial Ecology in Pig Manure

    Science.gov (United States)

    Cho, Sungback; Hwang, Okhwa; Park, Sungkwon

    2015-01-01

    This study was performed to investigate the effect of different levels of dietary crude protein (CP) on composition of odorous compounds and bacterial communities in pig manure. A total of 48 male pigs (average initial body weight 45 kg) fed diets containing three levels of dietary CP (20%, 17.5%, and 15%) and their slurry samples were collected from the pits under the floor every week for one month. Changes in composition of odorous compounds and bacterial communities were analyzed by gas chromatography and 454 FLX titanium pyrosequencing systems, respectively. Levels of phenols, indoles, short chain fatty acid and branched chain fatty acid were lowest (pacid, and iso-valeric acid with Atopostipes, p-cresol and skatole with Bacteroides, acetic acid and butyric acid with AM982595_g of Porphyromonadaceae family, and propionic acid with Tissierella. Taken together, administration of 15% CP showed less production of odorous compounds than 20% CP group and this result might be associated with the changes in bacterial communities especially whose roles in protein metabolism. PMID:26194219

  6. The Evaluation of Increase in Hemodialysis Frequency on C-Reactive Protein Levels and Nutritional Status

    Directory of Open Access Journals (Sweden)

    Shokooh Sarbolouki

    2013-02-01

    Full Text Available Malnutrition and inflammation are the most important causes of cardiovascular disease in hemodialysis patients. This study was conducted to evaluate the effect of increase in hemodialysis frequency on C-reactive protein (CRP level and nutritional markers in contrast to previous routine method. 18 hemodialysis patients with a mean age of 53±16 years were randomly selected in this before-and-after clinical trial. The patients under a standard hemodialysis of 3 times/4 h per week were converted to 4 times/4 h for a period of 6 weeks. The CRP, albumin, triglyceride, total cholesterol, LDL, HDL serum levels, anthropometric indices and 24-h diet recall intake was assessed before and after of the period. The data were analyzed using paired t-test, and P-value less than 0.05 was considered significant. All patients completed the study. Mean weight, body mass index and serum albumin increased while serum CRP level decreased significantly after the intervention (P<0.03. Triglyceride, total cholesterol, LDL, HDL, as well as energy, protein and fat intake had no significant change before and after the study. Increase in dialysis frequency decreased systemic inflammation and improved the nutritional state of hemodialysis patients. Therefore, it may decrease the risk of cardiovascular events in these patients

  7. Increasing levels of crude protein in multiple supplements for grazing beef heifers in rainy season

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    Lívia Vieira de Barros

    2015-06-01

    Full Text Available The objective was to evaluate the effect of multiple supplements with differents levels of crude protein (CP or mineral supplements on the nutritional parameters and performance of beef heifers grazing Uruchloa decumbens in the rainy season. A complete random design was employed. The treatments were made up of increasing levels of CP in the multiple supplements and a control treatment (MM in which animals were offered only mineral mixture. Multiple supplements contained 17; 30; 43 and 56% of CP, for treatments CP17; CP30; CP43 and CP56, respectively. Average daily gain (ADG (g was 447.7; 554.6; 638.4; 587.9; 590.4, for treatments MM, CP17; CP30; CP43 and CP56, respectively. A quadratic effect of the levels of crude protein was found (p< 0.10 on ADG. A greater intake of dry matter (DM, organic matter (OM, CP, ether extract (EE, non-fibrous carbohydrates (NFC, total digestible nutrients (TDN, and digested dry matter (p< 0.10 was found in animals supplemented with multiple supplements. Multiple supplements increased the apparent digestibility coefficient of DM, CP, EE and NFC. Supply of multiple multiple supplements for heifers grazing in medium to high quality pastures in the rainy season improves the performance of the animals.

  8. Interleukin-6, desmosome and tight junction protein expression levels in reflux esophagitis-affected mucosa

    Institute of Scientific and Technical Information of China (English)

    Fei-Yue Li; Yan Li

    2009-01-01

    AIM: To investigate the correlation between the expression levels of interleukin (IL)-6 and proteins in tight junctions (TJs) in the esophageal mucosa of rats modeling different types of reflux esophagitis (RE), and the ability of aluminum phosphate to protect against RE-induced mucosal damage via these proteins. METHODS: Male SPF Wistar rats aged 56 d were divided randomly into acid RE, alkaline RE, mixed RE, and control groups. Various surgical procedures were performed to establish rat models of acid RE. At 14 d after the procedure, some of the rats started aluminum phosphate treatment. Transmission electron microscopy (TEM) was used to observe the morphological features of TJs and desmosomes in the esophageal epithelium. Immunohistochemical methods and Western blotting were used to measure expression of claudin 1, occludin, ZO-1, JAM-1, DSG-1 and IL-6; reverse transcription polymerase chain reaction (RTPCR) was used to measure expression of mRNA of claudin 1, occludin, ZO-1, JAM-1, DSG-1 and IL-6. RESULTS: At day 14 after the procedures, an RE model was established in all subsequently sacrificed rats of groups A, B and C. By both gross and microscopic observation, the mucosa was damaged and thickened as the disease progressed. With TEM observation, a widened intercellular space was noticed, with significantly fewer desmosomes. Immunohistochemistry showed significantly higher levels of all proteins in all RE models compared to control rats at 3 d after operation (65.5% ± 25.6% vs 20.5% ± 2.1%, P 0.05, treated vs untreated, respectively). These levels increased in the rat with alkaline RE, and this increase was accompanied by continued hyperplasia in comparison with controls (85.5% ± 25.6% vs 20.5% ± 2.1%, P < 0.05, respectively). Furthermore, the expression of TJ proteins was not correlated significantly with that of IL-6 in this group. CONCLUSION: These findings indicate that TJ proteins are highly expressed as an early molecular event involved in RE

  9. Effect of Different Protein Levels On Nutrient Digestion Metabolism and Serum Biochemical Parameters in Calves

    Institute of Scientific and Technical Information of China (English)

    LI Hui; DIAO Qi-yu; ZHANG Nai-feng; TU Yan; WANG Ji-feng

    2008-01-01

    The current study has been performed to examine the effects of different dietary protein levels on the growth,nutrient digestion and absorption,as well as biochemical parameters in calves.Nine healthy newborn calves were selected,randomly divided into 3 groups and fed 3 milk replacers with different protein levels(18,22,and 26%),respectively.Five period-digestion-metabolism trials were taken between 12-20,22-30,32-40,42-50,and 52-60 days after birth.All 3 groups showed a similar growth curve during 11 to 61 experimental days,however,the growth rate of the 22%crude protein(CP) group was 8.89%higher than that of the 18%CP group and 19.48%higher than that of the 26%CP group.respectively. The apparent digestibility of dry material(DM)declined gradually with age,whereas,the apparent digestibility of N,extract ether(EE)rose slightly.Compared to the 22%CP and 26%CP group,calves fed with 18%CP apparently had a lower digestibility than DM,EE,and nitrogen(N).The average apparent digestibilities of N were 69.39,75.36,and 74.55%, respectively.Both the apparent digestibility and retention of calcium(Ca)and phosphorus(P)were steady throughout the experiment,but the average apparent digestibility of P in the 26%CP group was only 63.83%,which was markedly lower than that of the 18%CP group(70.40%)and 22%CP group(69.73%).In addition,the sernm concentrations of total protein(TP),albumin(ALB),and globulin(GLOB)of the 22%CP group were higher than those in the 18%CP and 26%CP groups.The urea N(BUN)content in the 18%CP group,on the other hand,was significantly lower than that of the other two groups.The highest glucose(GLU)concentration was found in the 22%CP group(5.38 mmol L-1),at the end of the trials.The protein levels in the milk replacer affected the digestion metabolism of nutrition and the serum biochemical parameters of calves at different physiological phases.Calves fed with 22%CP milk replacer had a better growth performance than the other groups.

  10. Improved cycling performance with ingestion of hydrolyzed marine protein depends on performance level

    Directory of Open Access Journals (Sweden)

    Vegge Geir

    2012-04-01

    Full Text Available Abstract Background The effect on performance of protein ingestion during or after exercise is not clear. This has largely been attributed to the utilization of different scientific protocols and the neglection of accounting for factors such as differences in physical and chemical properties of protein supplements and differences in athletic performance level. Methods We hypothesized that ingestion of unprocessed whey protein (15.3 g·h-1 together with carbohydrate (60 g·h-1, would provide no ergogenic effect on 5-min mean-power performance following 120 min cycling at 50% of maximal aerobic power (2.8 ± 0.2 W·kg-1, corresponding to 60 ± 4% of VO2max, compared to CHO alone (60 g·h-1. Conversely, we hypothesized that ingestion of the hydrolyzed marine protein supplement NutriPeptin™ (Np, 2.7 g·h-1, a processed protein supplement with potentially beneficial amino acid composition, together with a PROCHO beverage (12.4 g·h-1 and 60 g·h-1, respectively would provide an ergogenic effect on mean-power performance. We also hypothesized that the magnitude of the ergogenic effect of NpPROCHO would be dependent on athletic performance. As for the latter analysis, performance level was defined according to a performance factor, calculated from individual pre values of Wmax, VO2max and 5-min mean-power performance, wherein the performance of each subject was ranked relative to the superior cyclist whos performance was set to one. Twelve trained male cyclists (VO2max = 65 ± 4 ml·kg-1·min-1 participated in a randomized double-blinded cross-over study. Results and conclusions Overall, no differences were found in 5-min mean-power performance between either of the beverages (CHO 5.4 ± 0.5 W·kg-1; PROCHO 5.3 ± 0.5 W·kg-1; NpPROCHO 5.4 ± 0.3 W·kg-1 (P = 0.29. A negative correlation was found between NpPROCHO mean-power performance and athletic performance level (using CHO-performance as reference; Pearson R = -0.74, P = 0.006. Moreover

  11. Nutrient digestibility and growth of five breeds of sheep under different levels of undegradable protein

    OpenAIRE

    Yulistiani D; Naufaliah N; Kardaya D; Subandriyo

    2015-01-01

    A study was conducted to evaluate the effect of feeding different levels of rumen undegradable protein (UDP) in concentrate on performance of five breeds of sheep. Namely: Compass Agrinak (CA); Garut Composite breed (KG); Barbados Cross breed (BC); St Croix breed (SC); and Local Garut (GL) breed. Ten heads of sheep were used from each breed, in which each breed was grouped into 5 groups according to their body weight, and each sheep in each group was assigned to one of two treatments diet. Th...

  12. In vitro effect of dietary protein level and nondigestible oligosaccharides on feline fecal microbiota.

    Science.gov (United States)

    Pinna, C; Stefanelli, C; Biagi, G

    2014-12-01

    The aim of the present study was to evaluate in vitro the effect of some prebiotic substances and 2 dietary protein levels on the composition and activity of feline fecal microbiota. Two in vitro studies were conducted. First, 6 nondigestible oligosaccharides were studied; treatments were control diet (CTRL), gluconic acid (GA), carrot fiber (CF), fructooligosaccharides (FOS), galactooligosaccharides (GOS), lactitol (LAC), and pectins from citrus fruit (PEC). Substrates were added to feline fecal cultures at 2 g/L for 24 h incubation. Compared with the CTRL, ammonia had been reduced (P<0.05) by GOS (-9%) after 6 h and by GA (-14%), LAC (-12%), and PEC (-10%) after 24 h. After 24 h, all treatments had resulted in a lower pH versus the CTRL. Putrescine concentrations at 24 h were greater (P<0.05) in cultures treated with FOS (+90%), GOS (+96%), and LAC (+87%). Compared with the CTRL, total VFA were higher (P<0.05) in bottles containing CF (+41%), whereas the acetic to propionic acid ratio was reduced by LAC (-51%; P<0.05). After 24 h, Enterobacteriaceae had been reduced (P<0.05) by LAC and PEC. In a second study, LAC and FOS were selected to be tested in the presence of 2 diets differing in their protein content. There were 6 treatments: low-protein (LP) CTRL with no addition of prebiotics (CTRL-LP), high-protein (HP) CTRL with no addition of prebiotics (CTRL-HP), LP diet plus FOS, CTRL-HP plus FOS, LP diet plus LAC, and CTRL-HP plus LAC. Both FOS and LAC were added to feline fecal cultures at 2 g/L for 24 h incubation. Ammonia at 24 h was affected (P<0.05) by the protein level (36.2 vs. 50.2 mmol/L for LP and HP, respectively). The CTRL-HPs resulted in a higher pH and increased concentrations of biogenic amines were found after 6 and 24 h of incubation (P<0.05); putrescine at 24 h showed an increase (P<0.05) in cultures treated with FOS. Total VFA were influenced (P<0.05) by the protein level (40.9 vs. 32.6 mmol/L for LP and HP, respectively). At 24 h, the CTRL

  13. Eosinophil cationic protein levels parallel coagulation activation in the blister fluid of patients with bullous pemphigoid.

    Science.gov (United States)

    Tedeschi, A; Marzano, A V; Lorini, M; Balice, Y; Cugno, M

    2015-04-01

    Bullous pemphigoid (BP) is an autoimmune blistering disease due to autoantibodies against two hemidesmosomal antigens, namely BP180 and BP230, and characterized by coagulation activation both at cutaneous and systemic levels. Skin-infiltrating eosinophils contribute to bulla formation and, upon activation, are supposed to initiate the coagulation cascade. The aim of this study was to investigate whether the activation of eosinophils and coagulation are linked in BP. We evaluated the correlation between eosinophil cationic protein (ECP) levels and concentrations of the prothrombotic markers F1 + 2 and D-dimer in blister fluid and blood samples of 30 BP patients. Thirty healthy subjects were used as normal controls. ECP, F1 + 2 and D-dimer plasma levels were significantly higher in BP patients than in normal subjects. A significant correlation was found between ECP plasma levels and blood eosinophil count (r = 0.54, P = 0.002). F1 + 2 plasma levels positively correlated with disease severity, expressed as the percentage of body surface area involved (r = 0.36, P = 0.048). A striking increase in ECP (288.8 ± 45.2 ng/mL), F1 + 2 (31 409.9 ± 2929.4 pmol/L) and D-dimer levels (342 798.3 ± 44 206 ng/mL) was found in blister fluid from BP patients. In blister fluid, ECP levels were significantly higher than in peripheral blood (P blister fluids from BP patients and correlate with markers of coagulation activation, supporting the view that eosinophils initiate the coagulation cascade at skin level. © 2014 European Academy of Dermatology and Venereology.

  14. Effects of High Estrogen Levels on Monocyte Chemoattractant Protein-1 and Wound Healing.

    Science.gov (United States)

    Plackett, Timothy P; Gregory, Meredith S; Kovacs, Elizabeth J

    2015-02-01

    Objective: Herein, we tested the effects of high levels of supplemental estrogen treatment on cutaneous wound healing. Approach: Female mice were implanted with a 17β-estradiol (E2) secreting pellet or placebo before receiving a full-thickness dermal excisional wound. Mice receiving the E2 pellet attained hormone levels that are comparable to those achieved during pregnancy. At 1, 3, and 5 days after injury, the dermal excision wound was examined for their histologic appearance, rate of closure, and chemokine levels. Results: Wound closure, assessed by percent reepithelialization, was slower in E2-treated mice relative to placebo (42.6%±6.6% vs. 70.0%±5.3%, respectively, 3 days after injury). In addition, there was a marked reduction in the subepithelial inflammatory infiltrate and granulation tissue in E2-treated mice relative to placebo. Wound levels of monocyte chemoattractant protein-1 (MCP-1) were increased by 3 days after injury and continued to rise at 5 days after injury in placebo-treated mice (p<0.01). By contrast, MCP-1 levels were significantly reduced at 3 and 5 days after injury in E2-treated mice relative to placebo-treated controls (p<0.01). This attenuation could be reversed by treatment with an estrogen receptor antagonist. Innovation: High levels of estrogen are able to suppress normal wound closure. Conclusion: Dermal wound healing can be altered by manipulating the gonadal steroid hormone levels. In particular, high levels of estrogen can be utilized to slow down the rate of wound healing through a reduction in the inflammatory response.

  15. The Prognostic Value of C-Reactive Protein Serum Levels in Patients with Uterine Leiomyosarcoma.

    Directory of Open Access Journals (Sweden)

    Richard Schwameis

    Full Text Available C-reactive protein (CRP has previously been shown to serve as a prognostic parameter in women with gynecologic malignancies. Due to the lack of valid prognostic markers for uterine leiomyosarcoma (ULMS this study set out to investigate the value of pre-treatment CRP serum levels as prognostic parameter.Data of women with ULMS were extracted from databases of three Austrian centres for gynaecologic oncology. Pre-treatment CRP serum levels were measured and correlated with clinico-pathological parameters. Univariate and multivariable survival analyses were performed.In total, 53 patients with ULMS were included into the analysis. Mean (SD CRP serum level was 3.46 mg/dL (3.96. Solely, an association between pre-treatment CRP serum levels and tumor size (p = 0.04 but no other clinic-pathologic parameter such as tumor stage (p = 0.16, or histological grade (p = 0.07, was observed. Univariate and multivariable survival analyses revealed that CRP serum levels (HR 2.7 [1.1-7.2], p = 0.037 and tumor stage (HR 6.1 [1.9-19.5], p = 0.002 were the only independent prognostic factors for overall survival (OS in patients with ULMS. Patients with high pre-treatment CRP serum levels showed impaired OS compared to women with low levels (5-year-OS rates: 22.6% and 52.3%, p = 0.007.High pre-treatment CRP serum levels were independently associated with impaired prognosis in women with ULMS and might serve as a prognostic parameter in these patients.

  16. Effects of Two Different Levels of Dietary Protein on Body Composition and Protein Nutritional Status of Growing Rats

    Science.gov (United States)

    Tirapegui, Julio; Ribeiro, Sandra Maria Lima; Pires, Ivanir Santana de Oliveira; Rogero, Marcelo Macedo

    2012-01-01

    This study aimed to investigate the effect of a high-protein diet on growth, body composition, and protein nutritional status of young rats. Newly-weaned Wistar rats, weighing 45–50 g, were distributed in two experimental groups, according to their diets, which contained 12% (G12) or 26% protein (G26), over a period of 3 weeks. The animals were euthanized at the end of this period and the following analyses were performed: chemical composition of the carcass, proteoglycan synthesis, IGF-I concentration (serum, muscle and cartilage), total tissue RNA, protein concentration (muscle and cartilage) and protein synthesis (muscle and cartilage). The high-protein diet was found to result in a higher fat-free mass and lower fat mass in the carcass, with no difference in growth or protein nutritional status. PMID:23112920

  17. Effects of two different levels of dietary protein on body composition and protein nutritional status of growing rats.

    Science.gov (United States)

    Tirapegui, Julio; Ribeiro, Sandra Maria Lima; Pires, Ivanir Santana de Oliveira; Rogero, Marcelo Macedo

    2012-09-01

    This study aimed to investigate the effect of a high-protein diet on growth, body composition, and protein nutritional status of young rats. Newly-weaned Wistar rats, weighing 45-50 g, were distributed in two experimental groups, according to their diets, which contained 12% (G12) or 26% protein (G26), over a period of 3 weeks. The animals were euthanized at the end of this period and the following analyses were performed: chemical composition of the carcass, proteoglycan synthesis, IGF-I concentration (serum, muscle and cartilage), total tissue RNA, protein concentration (muscle and cartilage) and protein synthesis (muscle and cartilage). The high-protein diet was found to result in a higher fat-free mass and lower fat mass in the carcass, with no difference in growth or protein nutritional status.

  18. Stathmin protein level, a potential predictive marker for taxane treatment response in endometrial cancer.

    Science.gov (United States)

    Werner, Henrica M J; Trovik, Jone; Halle, Mari K; Wik, Elisabeth; Akslen, Lars A; Birkeland, Even; Bredholt, Therese; Tangen, Ingvild L; Krakstad, Camilla; Salvesen, Helga B

    2014-01-01

    Stathmin is a prognostic marker in many cancers, including endometrial cancer. Preclinical studies, predominantly in breast cancer, have suggested that stathmin may additionally be a predictive marker for response to paclitaxel. We first evaluated the response to paclitaxel in endometrial cancer cell lines before and after stathmin knock-down. Subsequently we investigated the clinical response to paclitaxel containing chemotherapy in metastatic endometrial cancer in relation to stathmin protein level in tumors. Stathmin level was also determined in metastatic lesions, analyzing changes in biomarker status on disease progression. Knock-down of stathmin improved sensitivity to paclitaxel in endometrial carcinoma cell lines with both naturally higher and lower sensitivity to paclitaxel. In clinical samples, high stathmin level was demonstrated to be associated with poor response to paclitaxel containing chemotherapy and to reduced disease specific survival only in patients treated with such combination. Stathmin level increased significantly from primary to metastatic lesions. This study suggests, supported by both preclinical and clinical data, that stathmin could be a predictive biomarker for response to paclitaxel treatment in endometrial cancer. Re-assessment of stathmin level in metastatic lesions prior to treatment start may be relevant. Also, validation in a randomized clinical trial will be important.

  19. Stathmin protein level, a potential predictive marker for taxane treatment response in endometrial cancer.

    Directory of Open Access Journals (Sweden)

    Henrica M J Werner

    Full Text Available Stathmin is a prognostic marker in many cancers, including endometrial cancer. Preclinical studies, predominantly in breast cancer, have suggested that stathmin may additionally be a predictive marker for response to paclitaxel. We first evaluated the response to paclitaxel in endometrial cancer cell lines before and after stathmin knock-down. Subsequently we investigated the clinical response to paclitaxel containing chemotherapy in metastatic endometrial cancer in relation to stathmin protein level in tumors. Stathmin level was also determined in metastatic lesions, analyzing changes in biomarker status on disease progression. Knock-down of stathmin improved sensitivity to paclitaxel in endometrial carcinoma cell lines with both naturally higher and lower sensitivity to paclitaxel. In clinical samples, high stathmin level was demonstrated to be associated with poor response to paclitaxel containing chemotherapy and to reduced disease specific survival only in patients treated with such combination. Stathmin level increased significantly from primary to metastatic lesions. This study suggests, supported by both preclinical and clinical data, that stathmin could be a predictive biomarker for response to paclitaxel treatment in endometrial cancer. Re-assessment of stathmin level in metastatic lesions prior to treatment start may be relevant. Also, validation in a randomized clinical trial will be important.

  20. C-reactive protein levels in schizophrenia: a review and meta-analysis.

    Science.gov (United States)

    Miller, Brian J; Culpepper, Nick; Rapaport, Mark H

    2014-01-01

    There is an impression in the literature that schizophrenia is associated with increased inflammation, including abnormal blood levels of the acute phase reactant C-reactive protein (CRP). We performed a meta-analysis of blood CRP levels to estimate the overall effect size, as well as a pooled analysis of the prevalence of an elevated CRP level in patients with schizophrenia and related disorders. We identified articles by searching PubMed, PsycInfo, and ISI, and the reference lists of identified studies. Eight studies met the inclusion criteria for the meta-analysis, and five studies were included in the pooled analysis. CRP levels were significantly increased in patients compared to controls (effect size=0.45, 95% confidence interval 0.34-0.55, pschizophrenia and related disorders. Our results support a growing body of literature that schizophrenia is associated with increased inflammation, although many studies did not control for potential confounding factors such as BMI and smoking. Given the high prevalence of elevated CRP, metabolic syndrome, and premature cardiovascular mortality, our findings also suggest that measurement of blood CRP levels may be germane to the clinical care of patients with schizophrenia and related disorders.

  1. Effects of different dietary of protein and lipid levels on the growth of freshwater prawns (Macrobrachium carcinus) broodstock

    OpenAIRE

    Mario Benítez-Mandujano; Jesús T. Ponce-Palafox

    2015-01-01

    ABSTRACTObjective. Evaluate the effects of varying dietary protein and lipid levels on the growth and body composition of adult freshwater prawns, Macrobrachium carcinus (Linnaeus 1758), in a recirculation system for 11 weeks (77 days). Materials and methods. The experimental treatments were assigned in triplicate. Six test diets were formulated with three different protein levels (35, 40 and 45%) and two lipid levels (8 and 13%). Results. The highest survival rate, growth indices and feed ut...

  2. Effects of different dietary of protein and lipid levels on the growth of freshwater prawns (Macrobrachium carcinus) broodstock

    OpenAIRE

    Mario Benítez-Mandujano; Jesús T. Ponce-Palafox

    2014-01-01

    ABSTRACTObjective. Evaluate the effects of varying dietary protein and lipid levels on the growth and body composition of adult freshwater prawns, Macrobrachium carcinus (Linnaeus 1758), in a recirculation system for 11 weeks (77 days). Materials and methods. The experimental treatments were assigned in triplicate. Six test diets were formulated with three different protein levels (35, 40 and 45%) and two lipid levels (8 and 13%). Results. The highest survival rate, growth indices and feed ut...

  3. Effects of dietary protein levels during rearing and dietary energy levels during lay on body composition and reproduction in broiler breeder females

    NARCIS (Netherlands)

    Emous, van R.A.; Kwakkel, R.P.; Krimpen, van M.M.; Hendriks, W.H.

    2015-01-01

    A study with a 2 × 3 × 2 factorial arrangement was conducted to determine the effects of 2 dietary protein levels (high = CPh and low = CPl) during rearing, 3 dietary energy levels (3,000, MEh1; 2,800, MEs1; and 2,600, MEl1, kcal/kg AMEn, respectively) during the first phase of lay, and 2 dietary

  4. deconSTRUCT: general purpose protein database search on the substructure level.

    Science.gov (United States)

    Zhang, Zong Hong; Bharatham, Kavitha; Sherman, Westley A; Mihalek, Ivana

    2010-07-01

    deconSTRUCT webserver offers an interface to a protein database search engine, usable for a general purpose detection of similar protein (sub)structures. Initially, it deconstructs the query structure into its secondary structure elements (SSEs) and reassembles the match to the target by requiring a (tunable) degree of similarity in the direction and sequential order of SSEs. Hierarchical organization and judicious use of the information about protein structure enables deconSTRUCT to achieve the sensitivity and specificity of the established search engines at orders of magnitude increased speed, without tying up irretrievably the substructure information in the form of a hash. In a post-processing step, a match on the level of the backbone atoms is constructed. The results presented to the user consist of the list of the matched SSEs, the transformation matrix for rigid superposition of the structures and several ways of visualization, both downloadable and implemented as a web-browser plug-in. The server is available at http://epsf.bmad.bii.a-star.edu.sg/struct_server.html.

  5. CHANGES IN ENDOGENOUS GENE TRANSCRIPT AND PROTEIN LEVELS IN MAIZE PLANTS EXPRESSING THE SOYBEAN FERRITIN TRANSGENE

    Directory of Open Access Journals (Sweden)

    Milly N Kanobe

    2013-06-01

    Full Text Available Transgenic agricultural crops with increased nutritive value present prospects for contributing to public health. However, their acceptance is poor in many countries due to the perception that genetic modification may cause unintended effects on expression of native genes in the host plant. Here, we tested effects of soybean ferritin transgene (SoyFer1, M64337 on transcript and protein levels of endogenous genes in maize. Results showed that the transgene was successfully introduced and expressed in the maize seed endosperm. mRNA abundance of seven tested iron homeostasis genes and seed storage protein genes differed significantly between seed samples positive and negative for the transgene. The PCR negative samples had higher zein and total protein content compared to the positive samples. However, PCR positive samples had significantly higher concentrations of calcium, magnesium and iron. We have shown that the soybean ferritin transgene affected the expression of native iron homeostasis genes in the maize plant. These results underscore the importance of taking a holistic approach to the evaluation of transgenic events in target plants, comparing the transgenic plant to the untransformed controls.

  6. Fluctuations in species-level protein expression occur during element and nutrient cycling in the subsurface.

    Directory of Open Access Journals (Sweden)

    Michael J Wilkins

    Full Text Available While microbial activities in environmental systems play a key role in the utilization and cycling of essential elements and compounds, microbial activity and growth frequently fluctuates in response to environmental stimuli and perturbations. To investigate these fluctuations within a saturated aquifer system, we monitored a carbon-stimulated in situ Geobacter population while iron reduction was occurring, using 16S rRNA abundances and high-resolution tandem mass spectrometry proteome measurements. Following carbon amendment, 16S rRNA analysis of temporally separated samples revealed the rapid enrichment of Geobacter-like environmental strains with strong similarity to G. bemidjiensis. Tandem mass spectrometry proteomics measurements suggest high carbon flux through Geobacter respiratory pathways, and the synthesis of anapleurotic four carbon compounds from acetyl-CoA via pyruvate ferredoxin oxidoreductase activity. Across a 40-day period where Fe(III reduction was occurring, fluctuations in protein expression reflected changes in anabolic versus catabolic reactions, with increased levels of biosynthesis occurring soon after acetate arrival in the aquifer. In addition, localized shifts in nutrient limitation were inferred based on expression of nitrogenase enzymes and phosphate uptake proteins. These temporal data offer the first example of differing microbial protein expression associated with changing geochemical conditions in a subsurface environment.

  7. Ursolic acid regulates aging process through enhancing of metabolic sensor proteins level.

    Science.gov (United States)

    Bahrami, Soroush Alaghehband; Bakhtiari, Nuredin

    2016-08-01

    We previously reported that Ursolic Acid (UA) ameliorates skeletal muscle performance through satellite cells proliferation and cellular energy status. In studying the potential role of the hypothalamus in aging, we developed a strategy to pursue UA effects on the hypothalamus anti-aging proteins such as; SIRT1, SIRT6, PGC-1β and α-Klotho. In this study, we used a model of aging animals (C57BL/6). UA dissolved in Corn oil (20mg/ml) and then administrated (200mg/Kg i.p injection) to mice, twice daily for 7days. After treatment times, the mice perfused and the hypothalamus isolated for preparing of tissue to Immunofluorescence microscopy. The data illustrated that UA significantly increased SIRT1 (∼3.5±0.3 folds) and SIRT-6 (∼1.5±0.2 folds) proteins overexpression (Presults showed that UA enhanced α-Klotho (∼3.3±0.3) and PGC-1β (∼2.6±0.2 folds) proteins levels (PUA through enhancing of anti-aging biomarkers (SIRT1 and SIRT6) and PGC-1β in hypothalamus regulates aging-process and attenuates mitochondrial-related diseases. In regard to the key role of α-Klotho in aging, our data indicate that UA may be on the horizon to forestall diseases of aging.

  8. Data on early postoperative changes in aqueous monocyte chemoattractant protein-1 levels after phacoemulsification.

    Science.gov (United States)

    Kawai, Motofumi; Inoue, Toshihiro; Yoshida, Akitoshi; Tanihara, Hidenobu

    2016-12-01

    The data presented in this article are related to the research article entitled "Elevated levels of monocyte chemoattractant protein-1 in the aqueous humor after phacoemulsification" (M. Kawai, T. Inoue, M. Inatani, N. Tsuboi, K. Shobayashi, A. Matsukawa, A. Yoshida, H, 2012) [1]. The mean (±SE) aqueous MCP-1 levels (pg/ml) were 31.2±12.5, 1931.2±910.7, 2172.2±1015.7, 3315.4 ±1535.8, 3015.9 ±914.4, 2709.0 ±738.7, 72.8 ±26.9, and 207.1±62.9 at 0, 3, 6, 12, 24, 48, 168, and 720 h after phacoemulsification, respectively. The immunohistochemical analysis showed a number of MCP-1 positive inflammatory cells in the anterior chamber and conjunctiva. There were some MCP-1 positive cells in the corneal endothelium.

  9. Relationship between high-sensitivity C-reactive protein level and angiographical characteristics of coronary atherosclerosis

    Institute of Scientific and Technical Information of China (English)

    JIA En-zhi; HUANG Jun; MA Wen-zhu; YANG Zhi-jian; YUAN Biao; ZANG Xiao-ling; WANG Rong-hu; ZHU Tie-bing; WANG Lian-sheng; CHEN Bo; CAO Ke-jiang

    2006-01-01

    @@ Arole for inflammation has become well established over the past decade or more in theories describing the atherosclerotic disease process.1,2 From a pathological viewpoint, all stages, ie, initiation, growth, and complication of the atherosclerotic plaque,3,4 might be considered to be an inflammatory response to injury. Several prospective studies 5-7 recently showed that plasma high sensitivity C-reactive protein (hsCRP) levels, which are one of the markers of systemic inflammation, are a powerful predictor of future myocardial infarction and cardiac death among apparently healthy individuals. However, the association between the plasma hsCRP levels and the extent of coronary stenosis in subjects remains controversial. Some studies previously demon- strated such associations,8,9 whereas other could not found.10,11 Gensini's score assigns a severity score for a stenosed vessel depending on the degree of luminal narrowing and the importance of its location.12

  10. Increased CSF levels of phosphorylated neurofilament heavy protein following bout in amateur boxers.

    Directory of Open Access Journals (Sweden)

    Sanna Neselius

    Full Text Available INTRODUCTION: Diagnosis of mild TBI is hampered by the lack of imaging or biochemical measurements for identifying or quantifying mild TBI in a clinical setting. We have previously shown increased biomarker levels of protein reflecting axonal (neurofilament light protein and tau and glial (GFAP and S-100B damage in cerebrospinal fluid (CSF after a boxing bout. The aims of this study were to find other biomarkers of mild TBI, which may help clinicians diagnose and monitor mild TBI, and to calculate the role of APOE ε4 allele genotype which has been associated with poor outcome after TBI. MATERIALS AND METHODS: Thirty amateur boxers with a minimum of 45 bouts and 25 non-boxing matched controls were included in a prospective cohort study. CSF and blood were collected at one occasion between 1 and 6 days after a bout, and after a rest period for at least 14 days (follow up. The controls were tested once. CSF levels of neurofilament heavy (pNFH, amyloid precursor proteins (sAPPα and sAPPβ, ApoE and ApoA1 were analyzed. In blood, plasma levels of Aβ42 and ApoE genotype were analyzed. RESULTS: CSF levels of pNFH were significantly increased between 1 and 6 days after boxing as compared with controls (p<0.001. The concentrations decreased at follow up but were still significantly increased compared to controls (p = 0.018. CSF pNFH concentrations correlated with NFL (r =  0.57 after bout and 0.64 at follow up, p<0.001. No significant change was found in the other biomarkers, as compared to controls. Boxers carrying the APOE ε4 allele had similar biomarker concentrations as non-carriers. CONCLUSIONS: Subconcussive repetitive trauma in amateur boxing causes a mild TBI that may be diagnosed by CSF analysis of pNFH, even without unconsciousness or concussion symptoms. Possession of the APOE ε4 allele was not found to influence biomarker levels after acute TBI.

  11. Ck2-Dependent Phosphorylation Is Required to Maintain Pax7 Protein Levels in Proliferating Muscle Progenitors.

    Directory of Open Access Journals (Sweden)

    Natalia González

    Full Text Available Skeletal muscle regeneration and long term maintenance is directly link to the balance between self-renewal and differentiation of resident adult stem cells known as satellite cells. In turn, satellite cell fate is influenced by a functional interaction between the transcription factor Pax7 and members of the MyoD family of muscle regulatory factors. Thus, changes in the Pax7-to-MyoD protein ratio may act as a molecular rheostat fine-tuning acquisition of lineage identity while preventing precocious terminal differentiation. Pax7 is expressed in quiescent and proliferating satellite cells, while its levels decrease sharply in differentiating progenitors Pax7 is maintained in cells (reacquiring quiescence. While the mechanisms regulating Pax7 levels based on differentiation status are not well understood, we have recently described that Pax7 levels are directly regulated by the ubiquitin-ligase Nedd4, thus promoting proteasome-dependent Pax7 degradation in differentiating satellite cells. Here we show that Pax7 levels are maintained in proliferating muscle progenitors by a mechanism involving casein kinase 2-dependent Pax7 phosphorylation at S201. Point mutations preventing S201 phosphorylation or casein kinase 2 inhibition result in decreased Pax7 protein in proliferating muscle progenitors. Accordingly, this correlates directly with increased Pax7 ubiquitination. Finally, Pax7 down regulation induced by casein kinase 2 inhibition results in precocious myogenic induction, indicating early commitment to terminal differentiation. These observations highlight the critical role of post translational regulation of Pax7 as a molecular switch controlling muscle progenitor fate.

  12. Protein- and tryptophan-restricted diets induce changes in rat gonadal hormone levels.

    Science.gov (United States)

    Del Angel-Meza, A R.; Feria-Velasco, A; Ontiveros-Martínez, L; Gallardo, L; Gonzalez-Burgos, I; Beas-Zárate, C

    2001-04-01

    The release of gonadotrophic hormones starts at puberty and, along with the subsequent estral cyclicity, is subject to hormonal feedback systems and to the action of diverse neuroactive substances such as gamma amino butyric acid and catecholamines. This study shows the effect of the administration during 40 days of protein-restricted and corn-based (tryptophan- and lysine-deficient) diets on the serotonin concentration in medial hypothalamic fragments as well as in follicle-stimulating luteinizing hormones, 17-beta-estradiol and progesterone serum levels, and estral cyclicity in 60- and 100-day-old rats (young, mature, and in gestation). In young rats, a delay in vaginal aperture development, and a lengthening of the estral cycle to a continuous anestral state was observed, mainly in the group fed corn. This group showed a 25% decrease in the serotonin concentration compared with the protein-restricted group, which exhibited an increase of 9% over the control group. Luteinizing hormone levels decreased in 16% and 13%, whereas follicle-stimulating hormone increased in 13% and 5% in the young animals of restricted groups, respectively, compared with the control group. Serum progesterone levels decreased only in young restricted versus control animals, and no differences were seen among adult and gestational rats. Serum levels of 17-beta-estradiol in restricted animals showed different concentration patterns, mainly in the corn group, which was higher at the 20th gestational day, falling drastically postpartum. The results obtained in this study show serotonin to be a very important factor in the release of gonadotrophic hormones and the start of puberty.

  13. Levels of procalcitonin, C-reactive protein and neopterin in patients with advanced HIV-1 infection

    Directory of Open Access Journals (Sweden)

    P Bipath

    2012-06-01

    Full Text Available Objectives. To compare the value of procalcitonin, C-reactive protein (CRP and neopterin as indicators of immune deficiency, co-infection, efficacy of treatment, and disease progression, in patients with advanced HIV-1 infection. Design. Cross-sectional, investigating baseline blood measurements and clinical observations in 82 HIV-positive patients divided into an antiretroviral treatment (ART group and an ART-naïve group. Setting. Secondary general hospital in Pretoria. Results. Procalcitonin and CRP levels showed no significant differences between the ART and ART-naïve groups, and no correlations with CD4 counts or viral loads. CRP levels were significantly higher with TB co-infection (p<0.05. Neopterin levels were raised above normal in 92% of the ART-naïve group and in 75% of the ART group. The levels were significantly higher (p<0.05 in the ART- naïve group. Negative correlations were found between neopterin and CD4 counts for the total patient group (r=-0.482; p<0.001. Neopterin was significantly (p<0.05 higher in the HIV/TB co-infection group than in those without TB. Higher neopterin levels at baseline were associated with a decline in CD4 counts over the ensuing 6-month period, and patients with higher baseline neopterin levels developed more complications over the 6-month period. Conclusions. Compared with procalcitonin and CRP, neopterin appears to be associated with the degree of immunodeficiency and of co-infection with TB. Neopterin levels may be investigated further as a measure of disease progression or treatment response. S Afr J HIV Med 2012;13(2:78-82.

  14. Novel E3 ubiquitin ligases that regulate histone protein levels in the budding yeast Saccharomyces cerevisiae.

    Directory of Open Access Journals (Sweden)

    Rakesh Kumar Singh

    Full Text Available Core histone proteins are essential for packaging the genomic DNA into chromatin in all eukaryotes. Since multiple genes encode these histone proteins, there is potential for generating more histones than what is required for chromatin assembly. The positively charged histones have a very high affinity for negatively charged molecules such as DNA, and any excess of histone proteins results in deleterious effects on genomic stability and cell viability. Hence, histone levels are known to be tightly regulated via transcriptional, posttranscriptional and posttranslational mechanisms. We have previously elucidated the posttranslational regulation of histone protein levels by the ubiquitin-proteasome pathway involving the E2 ubiquitin conjugating enzymes Ubc4/5 and the HECT (Homologous to E6-AP C-Terminus domain containing E3 ligase Tom1 in the budding yeast. Here we report the identification of four additional E3 ligases containing the RING (Really Interesting New Gene finger domains that are involved in the ubiquitylation and subsequent degradation of excess histones in yeast. These E3 ligases are Pep5, Snt2 as well as two previously uncharacterized Open Reading Frames (ORFs YKR017C and YDR266C that we have named Hel1 and Hel2 (for Histone E3 Ligases respectively. Mutants lacking these E3 ligases are sensitive to histone overexpression as they fail to degrade excess histones and accumulate high levels of endogenous histones on histone chaperones. Co-immunoprecipitation assays showed that these E3 ligases interact with the major E2 enzyme Ubc4 that is involved in the degradation related ubiquitylation of histones. Using mutagenesis we further demonstrate that the RING domains of Hel1, Hel2 and Snt2 are required for histone regulation. Lastly, mutants corresponding to Hel1, Hel2 and Pep5 are sensitive to replication inhibitors. Overall, our results highlight the importance of posttranslational histone regulatory mechanisms that employ multiple E3

  15. ENO1 Protein Levels in the Tumor Tissues and Circulating Plasma Samples of Non-small Cell Lung Cancer Patients

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    Ying ZHANG

    2010-12-01

    Full Text Available Background and objective Proper tumor markers are useful to diagnosis, prognosis and treatment for lung cancer. The aim of this study is to examine the levels of alpha-enolase (ENO1 protein in the tumor tissues and peripheral plasma samples obtained from non-small cell lung cancer (NSCLC patients, and evaluate its potential clinical significance. Methods The ENO1 protein levels in the tumor tissues and corresponding normal tissues from 16 cases of lung squamous cell carcinoma were analyzed by Western blot. The ENO1 protein levels in the plasma samples from 42 healthy individuals, 34 patients with lung benign disease and 84 patients with NSCLC were measured by double antibody sandwich enzyme-linked immunosorbent assay. Results For 87.5% (14/16 of the patients with lung squamous cell carcinoma, the ENO1 protein level in the tumor tissues was higher than that in the corresponding normal lung tissues. The ENO1 protein level in the plasma of NSCLC patients was significantly higher than that in the plasma of healthy individuals (P=0.031 and patients with lung benign disease (P=0.019. Furthermore, the ENO1 protein level was significantly higher in the plasma of patients with lung adenocarcinoma than that of patients with lung squamous cell carcinoma. Conclusion The elevated levels of ENO1 protein in the tumor tissues and the plasma samples from NSCLC patients indicate ENO1 may be a candidate biomarker of lung cancer.

  16. Zinc finger protein 521 overexpression increased transcript levels of Fndc5 in mouse embryonic stem cells

    Indian Academy of Sciences (India)

    Motahere-Sadat Hashemi; Abbas Kiani Esfahani; Maryam Peymani; Alireza Shoaraye Nejati; Kamran Ghaedi; Mohammad Hossein Nasr-Esfahani; Hossein Baharvand

    2016-03-01

    Zinc finger protein 521 is highly expressed in brain, neural stem cells and early progenitors of the human hematopoietic cells. Zfp521 triggers the cascade of neurogenesis inmouse embryonic stemcells through inducing expression of the early neuroectodermal genes Sox1, Sox3 and Pax6. Fndc5, a precursor of Irisin has inducing effects on the expression level of brain derived neurotrophic factor in hippocampus. Therefore, it is most likely that Fndc5 may play an important role in neural differentiation. To exhibit whether the expression of this protein is under regulation with Zfp521, we overexpressed Zfp521 in a stable transformants of mESCs expressing EGFP under control of Fndc5 promoter. Increased expression of Zfp521 enhanced transcription levels of both EGFP and endogenous Fndc5. This result was confirmed by overexpression the aforementioned vectors in HEK cells and indicated that Zfp521 functions upstream of Fndc5 expression. It is most likely that Zfp521 may act through the binding to its response element on Fndc5 core promoter. Therefore it is concluding that an enhanced expression of Fndc5 in neural progenitor cells is stimulated by Zfp521 overexpression in these cells.

  17. Proteotyping: A New Approach Studying Influenza Virus Evolution at the Protein Level

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Phylogenetic methods have been widely used to detect the evolution of influenza viruses.However, previous phylogenetic studies of influenza viruses do not make full use of the genetic information at the protein level and therefore cannot distinguish the subtle differences among viral genes. Proteotyping is a new approach to study influenza virus evolution. It aimed at mining the potential genetic information of the viral gene at the protein level by visualizing unique amino acid signatures (proteotypes). Neuraminidase gene fragments of some H5N1 avian influenza viruses were used as an example to illustrate how the proteotyping method worked. Bayesian analysis confirmed that the NA gene tree was mainly divided into three lineages. The NA proteotype analysis further suggested there might be multiple proteotypes within these three lineages and even within single genotypes. At the same time, some proteotypes might even involve more than one genotype. In particular, it also discovered some amino acids of viruses of some genotypes might co-reassort. All these results proved this approach could provide additional information in contrast to results from standard phylogenetic tree analysis.

  18. Blood parameters in growing pigs fed increasing levels of bacterial protein meal

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    Tauson Anne-Helene

    2007-11-01

    Full Text Available Abstract The experiment investigated the effects of increasing dietary levels of bacterial protein meal (BPM on various blood parameters reflecting protein and fat metabolism, liver function, and purine base metabolism in growing pigs. Sixteen barrows were allocated to four different experimental diets. The control diet was based on soybean meal. In the other three diets soybean meal was replaced with increasing levels of BPM, approximately 17%, 35%, and 50% of the nitrogen being derived from BPM. Blood samples from the jugular vein were taken when the body weights of the pigs were approximately 10 kg, 21 kg, 45 kg, and 77 kg. The blood parameters reflecting fat metabolism and liver function were not affected by diet. Both the plasma albumin and uric acid concentrations tended to decrease (P = 0.07 and 0.01, respectively with increasing dietary BPM content, whereas the plasma glucose concentration tended to increase (P = 0.07 with increasing dietary BPM content. It was concluded that up to 50% of the nitrogen could be derived from BPM without affecting metabolic function, as reflected in the measured blood parameters.

  19. Tricornered Kinase Regulates Synapse Development by Regulating the Levels of Wiskott-Aldrich Syndrome Protein.

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    Rajalaxmi Natarajan

    Full Text Available Precise regulation of synapses during development is essential to ensure accurate neural connectivity and function of nervous system. Many signaling pathways, including the mTOR (mechanical Target of Rapamycin pathway operate in neurons to maintain genetically determined number of synapses during development. mTOR, a kinase, is shared between two functionally distinct multi-protein complexes- mTORC1 and mTORC2, that act downstream of Tuberous Sclerosis Complex (TSC. We and others have suggested an important role for TSC in synapse development at the Drosophila neuromuscular junction (NMJ synapses. In addition, our data suggested that the regulation of the NMJ synapse numbers in Drosophila largely depends on signaling via mTORC2. In the present study, we further this observation by identifying Tricornered (Trc kinase, a serine/threonine kinase as a likely mediator of TSC signaling. trc genetically interacts with Tsc2 to regulate the number of synapses. In addition, Tsc2 and trc mutants exhibit a dramatic reduction in synaptic levels of WASP, an important regulator of actin polymerization. We show that Trc regulates the WASP levels largely, by regulating the transcription of WASP. Finally, we show that overexpression of WASP (Wiskott-Aldrich Syndrome Protein in trc mutants can suppress the increase in the number of synapses observed in trc mutants, suggesting that WASP regulates synapses downstream of Trc. Thus, our data provide a novel insight into how Trc may regulate the genetic program that controls the number of synapses during development.

  20. Maternal High Fat Diet Affects Offspring's Vitamin K-Dependent Proteins Expression Levels.

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    Stuart Lanham

    Full Text Available Studies suggest bone growth & development and susceptibility to vascular disease in later life are influenced by maternal nutrition, during intrauterine and early postnatal life. There is evidence for a role of vitamin K-dependent proteins (VKDPs including Osteocalcin, Matrix-gla protein, Periostin, and Gas6, in bone and vascular development. This study extends the analysis of VKDPs previously conducted in 6 week old offspring, into offspring of 30 weeks of age, to assess the longer term effects of a maternal and postnatal high fat (HF diet on VKDP expression. Overall a HF maternal diet and offspring diet exacerbated the bone changes observed. Sex specific and tissue specific differences were observed in VKDP expression for both aorta and femoral tissues. In addition, significant correlations were observed between femoral OCN, Periostin Gas6, and Vkor expression levels and measures of femoral bone structure. Furthermore, MGP, OCN, Ggcx and Vkor expression levels correlated to mass and fat volume, in both sexes. In summary the current study has highlighted the importance of the long-term effects of maternal nutrition on offspring bone development and the correlation of VKDPs to bone structure.

  1. The effect of protein-energy levels dietary on Kacang goats performances

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    MuchJi Martawidjaja

    1999-10-01

    Full Text Available An experiment was done to evaluate the protein-energy requirement for growing Kacang goats. Twelve males and 18 female goats, seven to eight months old were used in this study and randomized into three treatment groups, with four and six animals each, and were kept in individual pens. The treatments used were: R1= Elephant grass (E.G. + concentrate C1 (21% CP; 3.9 Mcal GE/kg, R2 = E.G. + concentrate C2 (17% CP; 3.7 Mcal GE/kg, and R3 = E.G. + concentrate C3 (12% CP; 3.5 Mcal GE/kg, respectively. Fresh Elephant grass was offered in restricted, and concentrate was offered at 3% of body weight. The experiment was carried out for 12 weeks. Data were analysed by using factorial completely randomized design 2x3 (3 rations and 2 sexes. Parameters measured were: feed intake; average daily gain and feed conversion. The results indicated that among treatments there was no significant difference on dry matter (DM and gross energy (GE intake (P>0.05, but crude protein (CP intake of R1 was 23,6% higher than treatment R2; treatment R2 was 38.1% higher than R3 (P0.05, but treatment R1 was 36.9% and significantly higher than R3 (P0.05, but ration R1 was more efficient than R3 (P0.05. It was concluded that protein intake and average daily gain were increased, and feed conversion was more efficient as the crude protein-energy levels increased in the ration. Feed intake and average daily gain of male goats were higher and feed conversion was more efficient than the female goats.

  2. Heme protein-induced tubular cytoresistance: expression at the plasma membrane level.

    Science.gov (United States)

    Zager, R A

    1995-05-01

    Following experimental rhabdomyolysis, animals become resistant to heme protein-induced acute renal failure (ARF). The goals of this study were to: (a) ascertain whether this resistance, previously documented only in vivo, is expressed directly at the proximal tubular cell level; (b) determine whether heme proteinuria (vs. other consequences of rhabdomyolysis) is its trigger; and (c) ascertain some of its subcellular determinants. Rats were injected with a borderline toxic dose of glycerol and 24 hours later proximal tubular segments (PTS) were isolated for study. Their vulnerability to diverse forms of injury (FeSO4-induced oxidant stress, hypoxia, Ca2+ ionophore, cytochalasin D, PLA2) was compared to that found in normal PTS. Post-glycerol PTS manifested significant resistance to each insult (decreased lactate dehydrogenase +/- N-acetyl-beta-D-glucosaminidase release). Protection against FeSO4 was virtually complete and it was associated with a 50% decrease in membrane lipid peroxidation. No decrease in hydroxyl radical generation was noted during the FeSO4 challenge (salicylate trap assessment), suggesting a primary increase in membrane resistance to attack. That PLA2 addition caused less deacylation, plasma membrane enzyme (alanine aminopeptidase) release, and LDH leakage from post-glycerol versus normal tubules supported this hypothesis. To test whether cytoresistance was specifically triggered by heme proteins (vs. being a non-specific filtered protein effect, or a result of endotoxin cascade activation), rats were injected with purified myoglobin, non-heme containing filterable proteins, or endotoxin. Only myoglobin induced cytoresistance. In vivo heme oxygenase inhibition (tin-protoporphyrin) did not block the emergence of cytoresistance and it was expressed despite Na,K-ATPase inhibition (ouabain) or cytoskeletal disruption (cytochalasin D). In vivo heat shock failed to protect. In conclusion, (1) rhabdomyolysis induces broad based proximal tubular

  3. How Integrated Management Strategies Promote Protein Quality of Cotton Embryos: High Levels of Soil Available N, N Assimilation and Protein Accumulation Rate.

    Science.gov (United States)

    Yang, HongKun; Meng, YaLi; Chen, BingLin; Zhang, XingYue; Wang, YouHua; Zhao, WenQing; Zhou, ZhiGuo

    2016-01-01

    Cottonseed is widely used as a source of ruminant feed and for industrial purposes. Therefore, there is a tremendous need to improve the nutritional value of cotton embryos. In this study, a conventional management (CM) and two integrated cotton management strategies (IMS1, IMS2) were performed at two soil fertility levels to study the relationships among soil N, N assimilation, embryonic protein accumulation and protein quality. The levels of proteins, essential amino acids, and semi-essential amino acids, especially those of glutamate, lysine, and methionine, were higher in IMS1 and IMS2 embryos than in CM embryos. These changes were significantly positively correlated with the soil-available N content, glutamine synthetase activity and peak value of protein accumulation rate and were negatively correlated with the free amino acid level. These results illustrated that integrated management strategies, especially the rates and timing of N application, raise the level of soil available N, which is beneficial for N assimilation in developing cotton embryos. The protein content was limited by the rate of protein accumulation rather than by the free amino acid content. The combination of target yield fertilization, a growth-driven N application schedule, a high plant density and the seedling raising with bio-organic fertilizer can substantially improve protein quality in cotton embryos, especially at a soil with low soil organic matter and total nitrogen.

  4. Effect of Dietary Protein Level and Origin on the Redox Status in the Digestive Tract of Mice

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    Guowei Le

    2008-04-01

    Full Text Available The present study was undertaken to evaluate the effects of high protein (soybean protein or casein on the balance between production of free radicals and antioxidant level in digestive organs of mice. For this purpose, male (C57BL/6J mice were adapted to experimental diets containing soybean protein or casein with 20% (normal protein diets, NPDs or 60% (high protein diets, HPDs, and HPDs supplemented with 0.06g/kg cysteamine. After two weeks of feeding, oxidative and antioxidative parameters in duodenum, liver and pancreas were measured. The results show that ingestion of high protein markedly increased contents of superoxide anion and malondialdehyde (MDA, decreased activities of superoxide dismutase (SOD, glutathione peroxidase (GSH-Px, catalase (CAT and Na+ K+-ATPase, and content of reduced glutathione (GSH in digestive organs of mice (P<0.05. Levels of oxidative parameters were lower and antioxidant capacity of both enzyme and non-enzyme was higher in mice fed with soybean protein than those fed with casein. In groups fed HPDs supplemented with cysteamine, oxidative stress was mitigated. However, oxidative parameter levels were still higher than those of NPD-fed groups. The present study indicates that ingestion of high protein diets could result in an imbalance between oxidant and antioxidant, and thus induce oxidative stress in digestive organs of mice. The oxidative damage was smaller in mice fed with high level of soy protein in comparison with casein.

  5. Effects of different levels of protein intake and physical training on growth and nutritional status of young rats.

    Science.gov (United States)

    Ribeiro, Sandra Maria Lima; Rogero, Marcelo Macedo; Bacurau, Reury Frank Pereira; de Campos, Patrícia Lopes; Luz, Silmara dos Santos; Lancha, Antonio Herber; Tirapegui, Julio

    2010-01-01

    This study aimed to investigate the effects of physical training, and different levels of protein intake in the diet, on the growth and nutritional status of growing rats. Newly-weaned Wistar rats (n=48) were distributed into six experimental groups; three of them were subjected to physical swim training (1 h per day, 5 d per week, for 4 wk, after 2 wk of familiarization) and the other three were considered as controls (non-trained). Each pair of groups, trained and non-trained, received diets with a different level of protein in their composition: 14%, 21% or 28%. The animals were euthanized at the end of the training period and the following analyses were performed: proteoglycan synthesis as a biomarker of bone and cartilage growth, IGF-I (insulin-like growth factor-I) assay as a biomarker of growth and nutritional status, total RNA and protein concentration and protein synthesis measured in vivo using a large-dose phenylalanine method. As a main finding, increased dietary protein, combined with physical training, was able to improve neither tissue protein synthesis nor muscle growth. In addition, cartilage and bone growth seem to be deteriorated by the lower and the higher levels of protein intake. Our data allow us to conclude that protein enhancement in the diet, combined with physical exercise, does not stimulate tissue protein synthesis or muscle mass growth. Furthermore, physical training, combined with low protein intake, was not favorable to bone development in growing animals.

  6. Increased Circulating Levels of Vitamin D Binding Protein in MS Patients

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    Arturo Ottavio Rinaldi

    2015-01-01

    Full Text Available Vitamin D (vitD low status is currently considered a main environmental factor in multiple sclerosis (MS etiology and pathogenesis. VitD and its metabolites are highly hydrophobic and circulate mostly bound to the vitamin D binding protein (DBP and with lower affinity to albumin, while less than 1% are in a free form. The aim of this study was to investigate whether the circulating levels of either of the two vitD plasma carriers and/or their relationship are altered in MS. We measured DBP and albumin plasma levels in 28 MS patients and 24 healthy controls. MS patients were found to have higher DBP levels than healthy subjects. Concomitant interferon beta therapy did not influence DBP concentration, and the difference with the control group was significant in both females and males. No significant correlation between DBP and albumin levels was observed either in healthy controls or in patients. These observations suggest the involvement of DBP in the patho-physiology of MS.

  7. Cholesteryl ester transfer protein levels and gene deficiency in Chinese patients with cardio-cerebrovascular diseases

    Institute of Scientific and Technical Information of China (English)

    庄一义; 汪俊军; 张宏娟; 李勇; 刘小传; 李露言; 陈光辉

    2002-01-01

    Objective To detect cholesteryl ester transfer protein (CETP) levels, frequencies of CETP D442G and Ⅰ14A mutations and characteristics of abnormal lipids in patients with cardio-cerebro vascular diseases. Methods Ninety-four myocardial infarction (MI) patients,110 stroke patients and 335 healthy controls were selected. The CETP concentration was determined using ELISA. The CETP activity was measured using a substrate of 14 C-radiolabeled discoidal bilayer particles. The CETP gene mutations were detected by PCR-RFLP. Results The CETP concentrations in the MI and stroke group, were higher than those in the controls. The gene mutation frequencies of D442G in the MI, stroke and control group were 3.5%, 3.6% and 5%, respectively, and the frequencies of Ⅰ14A were 1.05%, 0.91% and 1%, respectively. One case of D442G homozygote was detected in the healthy group. The frequency of two CETP gene mutations showed no significant difference among the patients and controls. The CETP concentration and activity in subjects with CETP mutations were one-third of those in the control group. The level of HDL-C, apo-A1 increased in the mutation subjects, while the TG level decreased. Conclusions The CETP level increased significantly in patients with cardio-cerebrovascular diseases. The carriers of CETP deficiency had CETP and lipid abnormalities.

  8. Metformin impairs systemic bile acid homeostasis through regulating SIRT1 protein levels.

    Science.gov (United States)

    Chen, Qi; Yang, Xiaoying; Zhang, Huabing; Kong, Xingxing; Yao, Lu; Cui, Xiaona; Zou, Yongkang; Fang, Fude; Yang, Jichun; Chang, Yongsheng

    2017-01-01

    Metformin is widely used to treat hyperglycemia. However, metformin treatment may induce intrahepatic cholestasis and liver injury in a few patients with type II diabetes through an unknown mechanism. Here we show that metformin decreases SIRT1 protein levels in primary hepatocytes and liver. Both metformin-treated wild-type C57 mice and hepatic SIRT1-mutant mice had increased hepatic and serum bile acid levels. However, metformin failed to change systemic bile acid levels in hepatic SIRT1-mutant mice. Molecular mechanism study indicates that SIRT1 directly interacts with and deacetylates Foxa2 to inhibit its transcriptional activity on expression of genes involved in bile acids synthesis and transport. Hepatic SIRT1 mutation elevates Foxa2 acetylation levels, which promotes Foxa2 binding to and activating genes involved in bile acids metabolism, impairing hepatic and systemic bile acid homeostasis. Our data clearly suggest that hepatic SIRT1 mediates metformin effects on systemic bile acid metabolism and modulation of SIRT1 activity in liver may be an attractive approach for treatment of bile acid-related diseases such as cholestasis. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. 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.

  10. EFFECT OF DECREASING DIETARY PROTEIN LEVELS WITH OPTIMAL AMINO ACIDS PROFILE ON THE PERFORMANCE OF BROILERS

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    Z. Kamran, M. Aslam Mirza, Ahsan-ul-Haq1 and S. Mahmood1

    2004-10-01

    Full Text Available A six-week trial was conducted to study the effect of decreasing dietary crude protein (CP level on the performance of broilers in hot climatic conditions. Four experimental rations having CP 23 (control group, 22, 21 and 20%, with optimal amino acid balance were prepared. All the four rations were isocaloric having ME 3200 kcal/kg with Energy: Protein (E:P 139.0, 146.5, 152.4 and 160 in diets A, B, C and D respectively. One hundred and twenty day-old chicks were randomly distributed into 12 experimental units, each having 10 chicks. Rations were randomly allotted to experimental units such that each unit received three replicates. The experimental diets were fed to birds from day 1st to 42nd. Performance of birds was monitored in terms of weight gain, feed consumption and feed conversion ratio (FCR. At the end of experiment, two birds per each replicate were randomly selected and slaughtered to record the data on carcass yield, breast meat yield, abdominal fat and composition of breast meat. Results of the trial suggested that weight gain was significantly (P<0.01 increased in birds on diets with CP 20 and 21%. Feed consumption and FCR remained un-changed for all the treatment groups. Eviscerated carcass yield was significantly (P<0.05 higher for the group fed on diet with 20% CP. Breast meat yield, abdominal fat and composition of breast meat also remained un-changed. Economic evaluation of the trial revealed that decreasing CP levels from 23 to 20% resulted in reduced feed cost per kg of live weight gain, which clearly indicated that this approach was useful especially in severe summer conditions. The overall picture of the study suggests that dietary protein level of broilers could be reduced from 23 to 20%, with beneficial effects on growth performance and carcass characteristics and increased economic returns in hot environmental conditions, provided that levels of essential amino acids are closely looked after.

  11. Elevated levels of Dickkopf-related protein 3 in seminal plasma of prostate cancer patients

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    Zenzmaier Christoph

    2011-11-01

    Full Text Available Abstract Background Expression of Dkk-3, a secreted putative tumor suppressor, is altered in age-related proliferative disorders of the human prostate. We now investigated the suitability of Dkk-3 as a diagnostic biomarker for prostate cancer (PCa in seminal plasma (SP. Methods SP samples were obtained from 81 patients prior to TRUS-guided prostate biopsies on the basis of elevated serum prostate-specific antigen (PSA; > 4 ng/mL levels and/or abnormal digital rectal examination. A sensitive indirect immunoenzymometric assay for Dkk-3 was developed and characterized in detail. SP Dkk-3 and PSA levels were determined and normalized to total SP protein. The diagnostic accuracies of single markers including serum PSA and multivariate models to discriminate patients with positive (N = 40 and negative (N = 41 biopsy findings were investigated. Results Biopsy-confirmed PCa showed significantly higher SP Dkk-3 levels (100.9 ± 12.3 vs. 69.2 ± 9.4 fmol/mg; p = 0.026. Diagnostic accuracy (AUC of SP Dkk-3 levels (0.633 was enhanced in multivariate models by including serum PSA (model A; AUC 0.658 or both, serum and SP PSA levels (model B; AUC 0.710. In a subpopulation with clinical follow-up > 3 years post-biopsy to ensure veracity of negative biopsy status (positive biopsy N = 21; negative biopsy N = 25 AUCs for SP Dkk-3, model A and B increased to 0.667, 0.724 and 0.777, respectively. Conclusions In multivariate models to detect PCa, inclusion of SP Dkk-3 levels, which were significantly elevated in biopsy-confirmed PCa patients, improved the diagnostic performance compared with serum PSA only.

  12. Association between serum levels of C-reactive protein and personality traits in women

    Directory of Open Access Journals (Sweden)

    Anckarsäter Henrik

    2008-04-01

    Full Text Available Abstract Background While low-grade inflammation has consistently been observed in subjects with depression, studies on the possible relationship between inflammation and other aspects of brain function are as yet sparse. In this study, we aimed to investigate the possible association between serum levels of the inflammation marker C-reactive protein (CRP and personality traits. Methods In this study, serum levels of high-sensitivity CRP were determined by ELISA in a population of 270 42-year-old women recruited from the population registry who had been assessed using the Temperament and Character Inventory. Self-reported previous or ongoing depression was also recorded. Unpaired two-tailed t-tests were used for comparison between two groups and correlations were evaluated by the calculation of Pearson's r-coefficient. Results The temperament trait harm avoidance was positively (r = 0.227, p r = -0.261, p p-values corrected for multiple comparisons. The correlations between the personality traits and CRP were observed also after exclusion of subjects reporting ongoing depression (n = 26. Whereas women reporting ongoing depression showed significantly increased levels of CRP as compared to non-depressed women (n = 155, women reporting a history of depression displayed no significant difference in CRP levels as compared to women that reported that they had never been depressed. Conclusion Serum levels of CRP in women was found to be associated with the personality traits harm avoidance and self-directedness. In addition, moderately elevated levels may be a state dependent marker of depression.

  13. PLASMA C-REACTIVE PROTEIN LEVELS AS A PROGNOSTIC MARKER IN FIRST EVER ACUTE ISCHEMIC STROKE

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    Bharat

    2014-12-01

    Full Text Available BACKGROUND AND PURPOSE: Acute ischemic stroke may trigger an inflammatory response that leads to increased levels of C-reactive protein (CRP. High levels of CRP may be associated with poor outcome because they reflect either an inflammatory reaction or tissue damage. We related plasma CRP levels to first ever ischemic stroke and its role as a diagnostic aid. METHODS: Sixty patients fulfilling inclusion and exclusion criteria with first ever acute ischemic stroke were included in study. CT scan of brain was done after 24 hours of onset of symptoms to confirm the diagnosis. Plasma CRP level was determined after 12 hours and before 72 hours of onset of symptoms in all CT confirmed ischemic stroke patients. This clinical study was done from January 2008 to June 2009. CRP was randomly measured in 60 age and sex matched individuals admitted in other wards of the hospital matched in all possible criteria expect the disease under study as a control group. RESULTS: The CRP concentration in ischemic strokes was independent of infarction site, the value was more between 51-70 years of age group and almost equal in both genders. 54 of the 60 ischemic strokes studied had CRP value >6 mg/l and only 6 patients had 6 mg/l, which is insignificant. CONCLUSION: The CRP level is significantly higher in ischemic strokes and by its elevation between 12-72 hours of symptom onset is a bad prognostic indicator. The risk of poor outcome or death at 3 months increased with higher levels of CRP. Elevated CRP values is a risk factor in association with other risk factors like diabetes/hypertension

  14. MAP17 and SGLT1 protein expression levels as prognostic markers for cervical tumor patient survival.

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    Marco Perez

    Full Text Available MAP17 is a membrane-associated protein that is overexpressed in human tumors. Because the expression of MAP17 increases reactive oxygen species (ROS generation through SGLT1 in cancer cells, in the present work, we investigated whether MAP17 and/or SGLT1 might be markers for the activity of treatments involving oxidative stress, such as cisplatin or radiotherapy. First, we confirmed transcriptional alterations in genes involved in the oxidative stress induced by MAP17 expression in HeLa cervical tumor cells and found that Hela cells expressing MAP17 were more sensitive to therapies that induce ROS than were parental cells. Furthermore, MAP17 increased glucose uptake through SGLT receptors. We then analyzed MAP17 and SGLT1 expression levels in cervical tumors treated with cisplatin plus radiotherapy and correlated the expression levels with patient survival. MAP17 and SGLT1 were expressed in approximately 70% and 50% of cervical tumors of different types, respectively, but they were not expressed in adenoma tumors. Furthermore, there was a significant correlation between MAP17 and SGLT1 expression levels. High levels of either MAP17 or SGLT1 correlated with improved patient survival after treatment. However, the patients with high levels of both MAP17 and SGLT1 survived through the end of this study. Therefore, the combination of high MAP17 and SGLT1 levels is a marker for good prognosis in patients with cervical tumors after cisplatin plus radiotherapy treatment. These results also suggest that the use of MAP17 and SGLT1 markers may identify patients who are likely to exhibit a better response to treatments that boost oxidative stress in other cancer types.

  15. Vitamin D receptor deletion leads to reduced level of IκBα protein through protein translation, protein-protein interaction, and post-translational modification

    OpenAIRE

    Wu, Shaoping; Xia, Yinglin; Liu, Xingyin; Sun, Jun

    2009-01-01

    Vitamin D receptor plays an essential role in regulation of inflammation. Previous studies demonstrate that vitamin D receptor negatively modulates the pro-inflammatory NF-κB pathway. However, it is unknown how vitamin D receptor regulates IκBα, the endogenous inhibitor of NF-κB. Here we investigated the molecular mechanism of vitamin D receptor deletion and IκBα expression. We found that cells lacking vitamin D receptor had significantly increased levels of IκBα mRNA and simultaneously decre...

  16. Effects of different dietary of protein and lipid levels on the growth of freshwater prawns (Macrobrachium carcinus broodstock

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    Mario Benítez-Mandujano

    2014-03-01

    Full Text Available Objective. Evaluate the effects of varying dietary protein and lipid levels on the growth and body composition of adult freshwater prawns, Macrobrachium carcinus (Linnaeus 1758, in a recirculation system for 11 weeks (77 days. Materials and methods. The experimental treatments were assigned in triplicate. Six test diets were formulated with three different protein levels (35, 40 and 45% and two lipid levels (8 and 13%. Results. The highest survival rate, growth indices and feed utilization were observed for M. carcinus adults fed protein:lipid diets of 35:13, 40:13 and 45:13, and the lowest values for these parameters were recorded for prawns fed diets with the lowest lipid levels; the differences in these parameters between these types of diets were significant (p<0.05. A nonsignificant tendency for an increased percentage of protein in the body with an increased dietary protein level was observed. The percentage of lipids decreased with an increasing dietary protein level, and no definite trends in ash content were found. Conclusions. The results suggest that a diet with 35% dietary crude protein and 13% lipids enhances the growth and body composition of adult M. carcinus.

  17. Increased plasma retinol binding protein 4 levels in patients with inflammatory cardiomyopathy.

    Science.gov (United States)

    Bobbert, Peter; Weithäuser, Alice; Andres, Janin; Bobbert, Thomas; Kühl, Uwe; Schultheiss, Heinz Peter; Rauch, Ursula; Skurk, Carsten

    2009-12-01

    Chronic heart failure (CHF) is associated with a higher risk for diabetes mellitus. Retinol binding protein 4 (RBP 4) is an adipose tissue-derived protein with pro-diabetogenic effects. A complete understanding of the association of CHF and insulin resistance remains elusive. The purpose of this study was to examine the relationship between CHF and diabetes mellitus. Plasma levels of RBP 4, insulin, and interleukins (IL) 2, 8, and 10, were assessed in patients with dilated cardiomyopathy (DCM, n = 53), dilated inflammatory cardiomyopathy (DCMi, n = 54), and controls (n = 20). In addition, a possible mechanism of RBP 4 regulation was examined in adipocytes in vitro. Plasma levels of RBP 4 and insulin were measured by a specific ELISA. Interleukin concentrations were obtained by multiplex ELISA. Cell culture with 3T3-L1 adipocytes was performed to measure RBP 4 mRNA expression after stimulation with IL-8. RBP 4 levels were significantly increased in patients with DCMi (52.95 +/- 20.42 microg/mL) compared with DCM (35.54 +/- 23.08 microg/mL) and the control group (27.3 +/- 18.51 microg/mL). RBP 4 was positively correlated with IL-8 (r=0.416, P < 0.05) in human plasma in patients with DCMi. Moreover, increased insulin resistance was observed in patients with DCMi compared with the control and DCM groups. In vitro, IL-8 induced a significant upregulation of RBP 4 mRNA expression in adipocytes. Elevated RBP 4 plasma concentrations, induced by IL-8, might be one mechanism leading to a higher incidence of diabetes in patients with DCMi.

  18. Effect of dietary protein quality and feeding level on milk secretion and mammary protein synthesis in the rat

    Energy Technology Data Exchange (ETDEWEB)

    Sampson, D.A.; Jansen, G.R.

    1985-04-01

    Protein synthesis was studied in mammary tissue of rats fed diets deficient in protein quality and/or restricted in food intake throughout gestation and lactation. Diets containing 25% wheat gluten (WG), wheat gluten plus lysine and threonine (WGLT), or casein (C) were pair-fed from conception until day 15 of lactation at 100% or 85% of WG ad libitum consumption (PF100 and PF85, respectively). A seventh group was fed C ad libitum. Rates of protein synthesis were measured in vivo at day 15 of lactation from incorporation of (3-/sup 3/H)phenylalanine. At both PF100 and PF85, fractional and absolute rates of mammary gland protein synthesis were two- to three-fold higher in rats fed C than in those fed WG. Pup weights showed similar treatment effects. Both mammary protein synthesis rates and pup weights were significantly higher in rats fed C at PF85 than rats fed WG ad libitum. Food restriction from PF100 to PF85 depressed pup weights and mammary protein synthesis rates in rats fed WGLT, but had no effect in rats fed WG. These results demonstrate that when food intake is restricted, improvement of protein quality of the maternal diet increases milk output in the rat in association with increased rates of mammary protein synthesis.

  19. Effects of Different Exercise Intensities with Isoenergetic Expenditures on C-Reactive Protein and Blood Lipid Levels

    Science.gov (United States)

    Tsao, Te Hung; Yang, Chang Bin; Hsu, Chin Hsing

    2012-01-01

    We investigated the effects of different exercise intensities on C-reactive protein (CRP), and whether changes in CRP levels correlated with blood lipid levels. Ten men exercised at 25%, 65%, and 85% of their maximum oxygen consumption rates. Participants' blood was analyzed for CRP and blood lipid levels before and after the exercise sessions.…

  20. Effects of Different Exercise Intensities with Isoenergetic Expenditures on C-Reactive Protein and Blood Lipid Levels

    Science.gov (United States)

    Tsao, Te Hung; Yang, Chang Bin; Hsu, Chin Hsing

    2012-01-01

    We investigated the effects of different exercise intensities on C-reactive protein (CRP), and whether changes in CRP levels correlated with blood lipid levels. Ten men exercised at 25%, 65%, and 85% of their maximum oxygen consumption rates. Participants' blood was analyzed for CRP and blood lipid levels before and after the exercise sessions.…

  1. Relation of resistin levels with C-reactve protein, homocysteine and uric acid in smokers and non-smokers

    Directory of Open Access Journals (Sweden)

    Onur Esbah

    2011-01-01

    Conclusions: We found that smoking may have influence on resistin levels and in smokers, insulin resistance is related to resistin levels, but in smoker and non-smokers body mass may not have any association with resistin. Resistin also may not have a role in C-reactive protein, homocysteine and uric acid levels both in smokers and non-smokers.

  2. Fish protein intake induces fast-muscle hypertrophy and reduces liver lipids and serum glucose levels in rats.

    Science.gov (United States)

    Kawabata, Fuminori; Mizushige, Takafumi; Uozumi, Keisuke; Hayamizu, Kohsuke; Han, Li; Tsuji, Tomoko; Kishida, Taro

    2015-01-01

    In our previous study, fish protein was proven to reduce serum lipids and body fat accumulation by skeletal muscle hypertrophy and enhancing basal energy expenditure in rats. In the present study, we examined the precise effects of fish protein intake on different skeletal muscle fiber types and metabolic gene expression of the muscle. Fish protein increased fast-twitch muscle weight, reduced liver triglycerides and serum glucose levels, compared with the casein diet after 6 or 8 weeks of feeding. Furthermore, fish protein upregulated the gene expressions of a fast-twitch muscle-type marker and a glucose transporter in the muscle. These results suggest that fish protein induces fast-muscle hypertrophy, and the enhancement of basal energy expenditure by muscle hypertrophy and the increase in muscle glucose uptake reduced liver lipids and serum glucose levels. The present results also imply that fish protein intake causes a slow-to-fast shift in muscle fiber type.

  3. Effects of Dietary Crude Protein Levels and Cysteamine Supplementation on Protein Synthetic and Degradative Signaling in Skeletal Muscle of Finishing Pigs.

    Directory of Open Access Journals (Sweden)

    Ping Zhou

    Full Text Available Dietary protein levels and cysteamine (CS supplementation can affect growth performance and protein metabolism of pigs. However, the influence of dietary protein intake on the growth response of CS-treated pigs is unclear, and the mechanisms involved in protein metabolism remain unknown. Hence, we investigated the interactions between dietary protein levels and CS supplementation and the effects of dietary crude protein levels and CS supplementation on protein synthetic and degradative signaling in skeletal muscle of finishing pigs. One hundred twenty barrows (65.84 ± 0.61 kg were allocated to a 2 × 2 factorial arrangement with five replicates of six pigs each. The primary variations were dietary crude protein (CP levels (14% or 10% and CS supplemental levels (0 or 700 mg/kg. The low-protein (LP diets (10% CP were supplemented with enough essential amino acids (EAA to meet the NRC AA requirements of pigs and maintain the balanced supply of eight EAA including lysine, methionine, threonine, tryptophan, valine, phenylalanine, isoleucine, and leucine. After 41 days, 10 pigs per treatment were slaughtered. We found that LP diets supplemented with EAA resulted in decreased concentrations of plasma somatostatin (SS (P<0.01 and plasma urea nitrogen (PUN (P<0.001, while dietary protein levels did not affect other traits. However, CS supplementation increased the average daily gain (P<0.001 and lean percentage (P<0.05, and decreased the feed conversion ratio (P<0.05 and back fat (P<0.05. CS supplementation also increased the concentrations of plasma insulin-like growth factor 1 (IGF-1 (P<0.001, and reduced the concentrations of leptin, SS, and PUN (P<0.001. Increased mRNA abundance of Akt1 and IGF-1 signaling (P<0.001 and decreased mRNA abundance of Forkhead Box O (FOXO 4 (P<0.01 and muscle atrophy F-box (P<0.001 were observed in pigs receiving CS. Additionally, CS supplementation increased the protein levels for the phosphorylated mammalian target of

  4. Effect of dietary energy levels and phase feeding by protein levels on growth performance, blood profiles and carcass characteristics in growing-finishing pigs.

    Science.gov (United States)

    Hong, J S; Lee, G I; Jin, X H; Kim, Y Y

    2016-01-01

    Providing of insufficient nutrients limits the potential growth of pig, while feeding of excessive nutrients increases the economic loss and causes environment pollution. For these reasons, phase feeding had been introduced in swine farm for improving animal production. This experiment was conducted to evaluate the effects of dietary energy levels and phase feeding by protein levels on growth performance, blood profiles and carcass characteristics in growing-finishing pigs. A total of 128 growing pigs ([Yorkshire × Landrace] × Duroc), averaging 26.62 ± 3.07 kg body weight, were assigned in a 2 × 4 factorial arrangement with 4 pigs per pen. The first factor was two dietary energy level (3,265 kcal of ME/kg or 3,365 kcal of ME/kg), and the second factor was four different levels of dietary protein by phase feeding (1growing(G)-2finishing(F) phases, 2G-2F phases, 2G-3F phases and 2G-3F phases with low CP requirement). In feeding trial, there was no significant difference in growth performance. The BUN concentration was decreased as dietary protein level decreased in 6 week and blood creatinine was increased in 13 week when pigs were fed diets with different dietary energy level. The digestibility of crude fat was improved as dietary energy levels increased and excretion of urinary nitrogen was reduced when low protein diet was provided. Chemical compositions of longissimus muscle were not affected by dietary treatments. In backfat thickness (P2) at 13 week, pigs fed high energy diet had thicker backfat thickness (P = 0.06) and pigs fed low protein diet showed the trend of backfat thinness reduction (P = 0.09). In addition, water holding capacity was decreased (P = 0.01) and cooking loss was increased (P = 0.07) as dietary protein level reduced. When pigs were fed high energy diet with low subdivision of phase feeding, days to 120 kg market weight was reached earlier compared to other treatments. Feeding the low energy diet

  5. Serum Heart-Type Fatty Acid-Binding Protein Levels in Patients with Overt Hypothyroidism

    Directory of Open Access Journals (Sweden)

    Esra Tutal

    2017-03-01

    Full Text Available Purpose: Overt hypothyroidism affects mostly women with an increasing prevalence with age. Hypothyroidism is associated with accelerated atherosclerotic cardiovascular diseases possibly caused by the higher incidence of hyperlipidemia, insulin resistance, and hypertension. Heart-type fatty acid-binding protein (H-FABP is specific for cardiomyocytes and a sensitive marker of myocardial injury. The purpose of this study was examining the effect of hypothyroidism on H-FABP levels and carotid artery intima-media thickness (CIMT. Material and Method: We measured serum H-FABP levels in 33 patients with overt hypothyroidism and age, gender, and body mass index-matched 39 control subjects. The patients were newly diagnosed with Hashimoto’s thyroiditis. All participants underwent high-resolution B-mode ultrasonography for the measurement of CIMT. Results: There was no significant difference in serum levels of H-FABP between the patient group and controls (1515.87±2143.0 pg/mL vs. 953.0±416.0 pg/mL, respectively; p=0.15. CIMT level was significantly higher in the patient group than in the control group (0.53±0.08 mm vs. 0.48±0.05 mm; p=0.02. However, the homeostasis model assessment of insulin resistance and fasting insulin levels did not differ between the two groups. Discussion: Based on the results of this study, we assume that H-FABP is not a useful marker in detecting preclinical atherosclerosis in patients with overt hypothyroidism associated with Hashimoto’s thyroiditis, however, CIMT might be a useful marker in detecting early atherosclerosis.

  6. Quantitative co-expression of proteins at the single cell level - application to a multimeric FRET sensor

    NARCIS (Netherlands)

    Goedhart, J.; van Weeren, L.; Adjobo-Hermans, M.J.W.; Elzenaar, I.; Hink, M.A.; Gadella (jr.), T.W.J.

    2011-01-01

    Background Co-expression of proteins is generally achieved by introducing two (or more) independent plasmids into cells, each driving the expression of a different protein of interest. However, the relative expression levels may vary strongly between individual cells and cannot be controlled. Ideall

  7. Quantitative co-expression of proteins at the single cell level--application to a multimeric FRET sensor.

    NARCIS (Netherlands)

    Goedhart, J.; Weeren, L. van; Adjobo-Hermans, M.J.W.; Elzenaar, I.; Hink, M.A.; Gadella, T.W.

    2011-01-01

    BACKGROUND: Co-expression of proteins is generally achieved by introducing two (or more) independent plasmids into cells, each driving the expression of a different protein of interest. However, the relative expression levels may vary strongly between individual cells and cannot be controlled. Ideal

  8. 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.

  9. A synthetic arabinose-inducible promoter confers high levels of recombinant protein expression in hyperthermophilic archaeon Sulfolobus islandicus

    DEFF Research Database (Denmark)

    Peng, Nan; Deng, Ling; Mei, Yuxia;

    2012-01-01

    levels of target gene expression. More strikingly, N-terminal amino acid sequencing of recombinant proteins unraveled that the protein synthesized from pEXA-N-lacS lacked the designed 6×His tag and that translation initiation did not start at the ATG codon of the fusion gene. Instead, it started...

  10. The human BDNF gene: peripheral gene expression and protein levels as biomarkers for psychiatric disorders

    Science.gov (United States)

    Cattaneo, A; Cattane, N; Begni, V; Pariante, C M; Riva, M A

    2016-01-01

    Brain-derived neurotrophic factor (BDNF) regulates the survival and growth of neurons, and influences synaptic efficiency and plasticity. The human BDNF gene consists of 11 exons, and distinct BDNF transcripts are produced through the use of alternative promoters and splicing events. The majority of the BDNF transcripts can be detected not only in the brain but also in the blood cells, although no study has yet investigated the differential expression of BDNF transcripts at the peripheral level. This review provides a description of the human BDNF gene structure as well as a summary of clinical and preclinical evidence supporting the role of BDNF in the pathogenesis of psychiatric disorders. We will discuss several mechanisms as possibly underlying BDNF modulation, including epigenetic mechanisms. We will also discuss the potential use of peripheral BDNF as a biomarker for psychiatric disorders, focusing on the factors that can influence BDNF gene expression and protein levels. Within this context, we have also characterized, for we believe the first time, the expression of BDNF transcripts in the blood, with the aim to provide novel insights into the molecular mechanisms and signaling that may regulate peripheral BDNF gene expression levels. PMID:27874848

  11. Cardiorespiratory fitness, pulmonary function and C-reactive protein levels in nonsmoking individuals with diabetes

    Energy Technology Data Exchange (ETDEWEB)

    Francisco, C.O.; Catai, A.M.; Moura-Tonello, S.C.G. [Universidade Federal de São Carlos, Departamento de Fisioterapia, São Carlos, SP, Brasil, Departamento de Fisioterapia, Universidade Federal de São Carlos, São Carlos, SP (Brazil); Lopes, S.L.B. [Universidade Federal de São Carlos, Departamento de Medicina, São Carlos, SP, Brasil, Departamento de Medicina, Universidade Federal de São Carlos, São Carlos, SP (Brazil); Benze, B.G. [Universidade Federal de São Carlos, Departamento de Estatística, São Carlos, SP, Brasil, Departamento de Estatística, Universidade Federal de São Carlos, São Carlos, SP (Brazil); Del Vale, A.M.; Leal, A.M.O. [Universidade Federal de São Carlos, Departamento de Medicina, São Carlos, SP, Brasil, Departamento de Medicina, Universidade Federal de São Carlos, São Carlos, SP (Brazil)

    2014-04-15

    The objective of this study was to evaluate cardiorespiratory fitness and pulmonary function and the relationship with metabolic variables and C-reactive protein (CRP) plasma levels in individuals with diabetes mellitus (DM). Nineteen men with diabetes and 19 age- and gender-matched control subjects were studied. All individuals were given incremental cardiopulmonary exercise and pulmonary function tests. In the exercise test, maximal workload (158.3±22.3 vs 135.1±25.2, P=0.005), peak heart rate (HR{sub peak}: 149±12 vs 139±10, P=0.009), peak oxygen uptake (VO{sub 2peak}: 24.2±3.2 vs 18.9±2.8, P<0.001), and anaerobic threshold (VO{sub 2VT}: 14.1±3.4 vs 12.2±2.2, P=0.04) were significantly lower in individuals with diabetes than in control subjects. Pulmonary function test parameters, blood pressure, lipid profile (triglycerides, HDL, LDL, and total cholesterol), and CRP plasma levels were not different in control subjects and individuals with DM. No correlations were observed between hemoglobin A1C (HbA1c), CRP and pulmonary function test and cardiopulmonary exercise test performance. In conclusion, the results demonstrate that nonsmoking individuals with DM have decreased cardiorespiratory fitness that is not correlated with resting pulmonary function parameters, HbA1c, and CRP plasma levels.

  12. C-reactive protein level and obesity as cardiovascular risk factors in polycystic ovary syndrome

    Directory of Open Access Journals (Sweden)

    Eda Ülkü Uludağ

    2013-09-01

    Full Text Available Objective: To investigate the role of C-reactive protein(CRP level elevation and obesity for the increased cardiovasculardisease risk in polycystic ovary syndrome(PCOS.Methods: A hundred and nine patients with PCOS and 30age matched healthy volunteers with regular menstrualcycle are involved in the study. PCOS group is furthersubdivided into three subgroups according to the bodymass index (BMI. Subgroups included 54 with BMI30. Blood samplesfor glucose, insulin, uric acid, and CRP were collected inthe morning after overnight fasting (12 hours. Homeostasismodel assessment-insulin resistance (HOMA-IRwas calculated. Results: Fasting blood glucose, insulin,and HOMA-IR was significantly higher in PCOS group(p=0.02, p=0.01 and p=0.02. CRP level was higher insubgroup with BMI>30. High CRP level in PCOS wasfound to be independent from BMI (p30.When compared with the control group high insulin levelwas the only to be statistically significant in obese PCOSpatients (p=0.005. HOMA-IR was higher in PCOS subgroupwith BMI>30 when compared with controls and thePCOS subgroup with BMI<25 (p<0.001, p= 0.003.Conclusion: Obesity, hyperinsulinemia, and high CRPlevels are seemed to be related and potentiating eachother in PCOS. Struggling with obesity is one of the mostimportant issues for preventive medicine.Key words: PCOS, CRP, obesity, cardiovascular risk

  13. Glucose regulates diacylglycerol intracellular levels and protein kinase C activity by modulating diacylglycerol kinase subcellular localization.

    Science.gov (United States)

    Miele, Claudia; Paturzo, Flora; Teperino, Raffaele; Sakane, Fumio; Fiory, Francesca; Oriente, Francesco; Ungaro, Paola; Valentino, Rossella; Beguinot, Francesco; Formisano, Pietro

    2007-11-02

    Although chronic hyperglycemia reduces insulin sensitivity and leads to impaired glucose utilization, short term exposure to high glucose causes cellular responses positively regulating its own metabolism. We show that exposure of L6 myotubes overexpressing human insulin receptors to 25 mm glucose for 5 min decreased the intracellular levels of diacylglycerol (DAG). This was paralleled by transient activation of diacylglycerol kinase (DGK) and of insulin receptor signaling. Following 30-min exposure, however, both DAG levels and DGK activity returned close to basal levels. Moreover, the acute effect of glucose on DAG removal was inhibited by >85% by the DGK inhibitor R59949. DGK inhibition was also accompanied by increased protein kinase C-alpha (PKCalpha) activity, reduced glucose-induced insulin receptor activation, and GLUT4 translocation. Glucose exposure transiently redistributed DGK isoforms alpha and delta, from the prevalent cytosolic localization to the plasma membrane fraction. However, antisense silencing of DGKdelta, but not of DGKalpha expression, was sufficient to prevent the effect of high glucose on PKCalpha activity, insulin receptor signaling, and glucose uptake. Thus, the short term exposure of skeletal muscle cells to glucose causes a rapid induction of DGK, followed by a reduction of PKCalpha activity and transactivation of the insulin receptor signaling. The latter may mediate, at least in part, glucose induction of its own metabolism.

  14. Hyperglycemic myocardial damage is mediated by proinflammatory cytokine: macrophage migration inhibitory factor.

    Directory of Open Access Journals (Sweden)

    Xi-Yong Yu

    Full Text Available BACKGROUND: Diabetes has been regarded as an inflammatory condition which is associated with left ventricular diastolic dysfunction (LVDD. The purpose of this study was to examine the expression levels of macrophage migration inhibitory factor (MIF and G protein-coupled receptor kinase 2 (GRK2 in patients with early diabetic cardiomyopathy, and to investigate the mechanisms involved in MIF expression and GRK2 activation. METHODS: 83 patients in the age range of 30-64 years with type 2 diabetes and 30 matched healthy men were recruited. Left ventricular diastolic function was evaluated by cardiac Doppler echocardiography. Plasma MIF levels were determined by ELISA. To confirm the clinical observation, we also studied MIF expression in prediabetic rats with impaired glucose tolerance (IGT and relationship between MIF and GRK2 expression in H9C2 cardiomyoblasts exposed to high glucose. RESULTS: Compared with healthy subjects, patients with diabetes have significantly increased levels of plasma MIF which was further increased in diabetic patients with Left ventricular diastolic dysfunction (LVDD. The increased plasma MIF levels in diabetic patients correlated with plasma glucose, glycosylated hemoglobin and urine albumin levels. We observed a significant number of TUNEL-positive cells in the myocardium of IGT-rats but not in the control rats. Moreover, we found higher MIF expression in the heart of IGT with cardiac dysfunction compared to that of the controls. In H9C2 cardiomyoblast cells, MIF and GRK2 expression was significantly increased in a glucose concentration-dependant manner. Furthermore, GRK2 expression was abolished by siRNA knockdown of MIF and by the inhibition of CXCR4 in H9C2 cells. CONCLUSIONS: Our findings indicate that hyperglycemia is a causal factor for increased levels of pro-inflammatory cytokine MIF which plays a role in the development of cardiomyopathy occurring in patients with type 2 diabetes. The elevated levels of MIF

  15. Identification of a functional SNP in the 3'-UTR of caprine MTHFR gene that is associated with milk protein levels.

    Science.gov (United States)

    An, Xiaopeng; Song, Yuxuan; Hou, Jinxing; Wang, Shan; Gao, Kexin; Cao, Binyun

    2016-08-01

    Xinong Saanen (n = 305) and Guanzhong (n = 317) dairy goats were used to detect SNPs in the caprine MTHFR 3'-UTR by DNA sequencing. One novel SNP (c.*2494G>A) was identified in the said region. Individuals with the AA genotype had greater milk protein levels than did those with the GG genotype at the c.*2494 G>A locus in both dairy goat breeds (P A substitution could increase the binding activity of bta-miR-370 with the MTHFR 3'-UTR. In addition, we observed a significant increase in the MTHFR protein level of AA carriers relative to that of GG carriers. These altered levels of MTHFR protein may account for the association of the SNP with milk protein level.

  16. The relationship between cholesteryl ester transfer protein levels and risk factor profile in patients with familial hypercholesterolemia

    NARCIS (Netherlands)

    de Grooth, Greetje J; Smilde, Tineke J; Van Wissen, Sanne; Klerkx, Anke H E M; Zwinderman, Aeilko H; Fruchart, Jean-Charles; Kastelein, John J P; Stalenhoef, Anton F H; Kuivenhoven, Jan Albert

    2004-01-01

    BACKGROUND: Cholesteryl ester transfer protein (CETP) mediates the transfer of neutral lipids between lipoproteins. The role of CETP in atherogenesis is controversial. To better understand the relationships between plasma CETP levels, lipoproteins and atherosclerosis, we assessed these parameters in

  17. Present perspectives on the automated classification of the G-protein coupled receptors (GPCRs) at the protein sequence level

    DEFF Research Database (Denmark)

    Davies, Matthew N; Gloriam, David E; Secker, Andrew;

    2011-01-01

    The G-protein coupled receptors--or GPCRs--comprise simultaneously one of the largest and one of the most multi-functional protein families known to modern-day molecular bioscience. From a drug discovery and pharmaceutical industry perspective, the GPCRs constitute one of the most commercially...... and economically important groups of proteins known. The GPCRs undertake numerous vital metabolic functions and interact with a hugely diverse range of small and large ligands. Many different methodologies have been developed to efficiently and accurately classify the GPCRs. These range from motif-based techniques...

  18. Insulin-like growth factor binding proteins increase intracellular calcium levels in two different cell lines.

    Directory of Open Access Journals (Sweden)

    Danielle Seurin

    Full Text Available BACKGROUND: Insulin-like growth factor binding proteins (IGFBPs are six related secreted proteins that share IGF-dependent and -independent functions. If the former functions begin to be well described, the latter are somewhat more difficult to investigate and to characterize. At the cellular level, IGFBPs were shown to modulate numerous processes including cell growth, differentiation and apoptosis. However, the molecular mechanisms implicated remain largely unknown. We previously demonstrated that IGFBP-3, but not IGFBP-1 or IGFBP-5, increase intracellular calcium concentration in MCF-7 cells (Ricort J-M et al. (2002 FEBS lett 527: 293-297. METHODOLOGY/PRINCIPAL FINDINGS: We perform a global analysis in which we studied, by two different approaches, the binding of each IGFBP isoform (i.e., IGFBP-1 to -6 to the surface of two different cellular models, MCF-7 breast adenocarcinoma cells and C2 myoblast proliferative cells, as well as the IGFBP-induced increase of intracellular calcium concentration. Using both confocal fluorescence microscopy and flow cytometry analysis, we showed that all IGFBPs bind to MCF-7 cell surface. By contrast, only four IGFBPs can bind to C2 cell surface since neither IGFBP-2 nor IGFBP-4 were detected. Among the six IGFBPs tested, only IGFBP-1 did not increased intracellular calcium concentration whatever the cellular model studied. By contrast, IGFBP-2, -3, -4 and -6, in MCF-7 cells, and IGFBP-3, -5 and -6, in C2 proliferative cells, induce a rapid and transient increase in intracellular free calcium concentration. Moreover, IGFBP-2 and -3 (in MCF-7 cells and IGFBP-5 (in C2 cells increase intracellular free calcium concentration by a pertussis toxin sensitive signaling pathway. CONCLUSIONS: Our results demonstrate that IGFBPs are able to bind to cell surface and increase intracellular calcium concentration. By characterizing the IGFBPs-induced cell responses and intracellular couplings, we highlight the cellular

  19. DRAM1 regulates apoptosis through increasing protein levels and lysosomal localization of BAX

    Science.gov (United States)

    Guan, J-J; Zhang, X-D; Sun, W; Qi, L; Wu, J-C; Qin, Z-H

    2015-01-01

    DRAM1 (DNA damage-regulated autophagy modulator 1) is a TP53 target gene that modulates autophagy and apoptosis. We previously found that DRAM1 increased autophagy flux by promoting lysosomal acidification and protease activation. However, the molecular mechanisms by which DRAM1 regulates apoptosis are not clearly defined. Here we report a novel pathway by which DRAM1 regulates apoptosis involving BAX and lysosomes. A549 or HeLa cells were treated with the mitochondrial complex II inhibitor, 3-nitropropionic acid (3NP), or an anticancer drug, doxorubicin. Changes in the protein and mRNA levels of BAX and DRAM1 and the role of DRAM1 in BAX induction were determined. The interaction between DRAM1 and BAX and its effect on BAX degradation, BAX lysosomal localization, the release of cathepsin B and cytochrome c by BAX and the role of BAX in 3NP- or doxorubicin-induced cell death were studied. The results showed that BAX, a proapoptotic protein, was induced by DRAM1 in a transcription-independent manner. BAX was degraded by autophagy under basal conditions; however, its degradation was inhibited when DRAM1 expression was induced. There was a protein interaction between DRAM1 and BAX and this interaction prolonged the half-life of BAX. Furthermore, upregulated DRAM1 recruited BAX to lysosomes, leading to the release of lysosomal cathepsin B and cleavage of BID (BH3-interacting domain death agonist). BAX mediated the release of mitochondrial cytochrome c, activation of caspase-3 and cell death partially through the lysosome-cathepsin B-tBid pathway. These results indicate that DRAM1 regulates apoptosis by inhibiting BAX degradation. In addition to mitochondria, lysosomes may also be involved in BAX-initiated apoptosis. PMID:25633293

  20. Expression levels of autophagy related proteins and their prognostic significance in retinocytoma and retinoblastoma

    Institute of Scientific and Technical Information of China (English)

    Yue; Liu; Shang-Tao; Wan; Ping; Zhang; Wen-Xin; Zhang; Jian-Ling; Zheng; Jian-Xian; Lin; Yong-Ping; Li

    2014-01-01

    ·AIM: To discuss the prognostic significant of autophagy related proteins(ARPs) in retinoblastoma(RB)and to find the molecular marker to distinguish retinocytoma(RC) and RB by investigating the different expression profiling of microtubule-associated protein light chain 3(LC3B) and other ARPs in RC and RB.·METHODS: Specimens with retinocytoma region(RCR)or mainly composed with Flexner-Winterstein rosettes(FWR) were screen out from 219 paraffin-embedded RB samples and respectively taken as RCR group and FWR group. Others were taken as undifferentiated(UD) group.Immunochemistry(IHC) of LC3 B and electronic microscopy was used to identify autophagy. The IHC scores of LC3 B and other ARPs, such as Beclin, PTEN,p27, p16INK4 a, mTOR and BCL-2 were compared and correlation analysis was applied to find potential proteins which may involve in autophagy regulation. The prognostics significance of LC3 B was evaluated by comparing the high risk features(HRFs) in 3 groups of total 219 samples.·RESULTS: Twenty-one specimens with RCR and 36 specimens mainly composed with FWR were screen out.RCR cell had a high level of LC3 B and lots of autophagic vacuoles. Beclin, PTEN, p27 had positive correlation with LC3, and p16INK4 ahad negative correlation, while the expression of mTOR and BCL-2 in RCR and RB region did not show any difference. Cases with RCR had lower rate of HRFs than undifferentiated cases.·CONCLUSION: ARPs had different expression pattern between RCR and other pathological types of RB, and could be ideal markers to distinguish RC from RB. Our finding indicated cases with RCR had favorable prognosis just like those with FWR.

  1. Training cessation does not alter GLUT-4 protein levels in human skeletal muscle.

    Science.gov (United States)

    Houmard, J A; Hortobágyi, T; Neufer, P D; Johns, R A; Fraser, D D; Israel, R G; Dohm, G L

    1993-02-01

    The purpose of this study was to determine whether short-term training cessation resulted in reduced GLUT-4 protein levels. Endurance- (n = 12, ET) and strength-trained (n = 12) individuals (ST) were examined before and after 14 days of training withdrawal. GLUT-4 content was determined from muscle biopsy samples of the gastrocnemius in ET and the vastus lateralis in ST. Insulin sensitivity (oral glucose tolerance test) was significantly (P 0.05) in both groups (92 and 100% of trained values for ET and ST, respectively). In ET, citrate synthase activity decreased significantly (P < 0.05) with training withdrawal (41.0 +/- 3.6 vs. 30.6 +/- 2.8 mumol.g-1.min-1); in ST no change was evident. The decrement in insulin sensitivity with the cessation of endurance- or resistance-oriented activity is therefore not associated with a reduction in GLUT-4 protein content. Muscle oxidative capacity and GLUT-4 content do not coincide with the removal of endurance training.

  2. ABI4 mediates antagonistic effects of abscisic acid and gibberellins at transcript and protein levels.

    Science.gov (United States)

    Shu, Kai; Chen, Qian; Wu, Yaorong; Liu, Ruijun; Zhang, Huawei; Wang, Pengfei; Li, Yanli; Wang, Shengfu; Tang, Sanyuan; Liu, Chunyan; Yang, Wenyu; Cao, Xiaofeng; Serino, Giovanna; Xie, Qi

    2016-02-01

    Abscisic acid (ABA) and gibberellins (GAs) are plant hormones which antagonistically mediate numerous physiological processes, and their optimal balance is essential for normal plant development. However, the molecular mechanism underlying ABA and GA antagonism still needs to be determined. Here, we report that ABA-INSENSITIVE 4 (ABI4) is a central factor in GA/ABA homeostasis and antagonism in post-germination stages. ABI4 overexpression in Arabidopsis (OE-ABI4) leads to developmental defects including a decrease in plant height and poor seed production. The transcription of a key ABA biosynthetic gene, NCED6, and of a key GA catabolic gene, GA2ox7, is significantly enhanced by ABI4 overexpression. ABI4 activates NCED6 and GA2ox7 transcription by directly binding to the promoters, and genetic analysis revealed that mutation in these two genes partially rescues the dwarf phenotype of ABI4 overexpressing plants. Consistently, ABI4 overexpressing seedlings have a lower GA/ABA ratio than the wild type. We further show that ABA induces GA2ox7 transcription while GA represses NCED6 expression in an ABI4-dependent manner; and that ABA stabilizes the ABI4 protein whereas GA promotes its degradation. Taken together, these results suggest that ABA and GA antagonize each other by oppositely acting on ABI4 transcript and protein levels.

  3. Accuracy of the FAMACHA© method in ewes fed different levels of crude protein

    Directory of Open Access Journals (Sweden)

    Francisco de Assis Fonseca de Macedo

    2014-05-01

    Full Text Available The accuracy of the FAMACHA© method was evaluated on the identification of female sheep fed two levels of crude protein, naturally infected with Haemonchus contortus, by means of the correspondent hematocrit value. Forty-seven female sheep of the breeds Santa Inês (n = 16, Texel (n = 16 and Ile de France (n = 15 aged between eight and twelve months were assigned to two treatments, 12 or 16 % crude protein in the diet. All the animals were wormed thirty days before the first data collections, which were done fortnightly between July 2005 and March 2006. The color of the ocular conjunctiva was individually evaluated according to the precepts of the FAMACHA© method and the hematocrit value of each animal was obtained in laboratory. A correlation of 1:0.7991 was found between the hematocrit values and the classification given by the FAMACHA© method aiming to identify animals with different degrees of anemia. The method was efficient to identify animals to worm, thus representing a support in the identification of animals susceptible to Haemonchus contortus.

  4. The Highwire ubiquitin ligase promotes axonal degeneration by tuning levels of Nmnat protein.

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    Xin Xiong

    Full Text Available Axonal degeneration is a hallmark of many neuropathies, neurodegenerative diseases, and injuries. Here, using a Drosophila injury model, we have identified a highly conserved E3 ubiquitin ligase, Highwire (Hiw, as an important regulator of axonal and synaptic degeneration. Mutations in hiw strongly inhibit Wallerian degeneration in multiple neuron types and developmental stages. This new phenotype is mediated by a new downstream target of Hiw: the NAD+ biosynthetic enzyme nicotinamide mononucleotide adenyltransferase (Nmnat, which acts in parallel to a previously known target of Hiw, the Wallenda dileucine zipper kinase (Wnd/DLK MAPKKK. Hiw promotes a rapid disappearance of Nmnat protein in the distal stump after injury. An increased level of Nmnat protein in hiw mutants is both required and sufficient to inhibit degeneration. Ectopically expressed mouse Nmnat2 is also subject to regulation by Hiw in distal axons and synapses. These findings implicate an important role for endogenous Nmnat and its regulation, via a conserved mechanism, in the initiation of axonal degeneration. Through independent regulation of Wnd/DLK, whose function is required for proximal axons to regenerate, Hiw plays a central role in coordinating both regenerative and degenerative responses to axonal injury.

  5. Variation of Serum Levels of Glucose and Protein During the Day in Sheep of Different Sex

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    Carmen Galván Doria

    2014-07-01

    Full Text Available In order to establish the effect of sampling hour and sex on blood glucose levels and total protein, there were monitored six Colombian native sheep for 3 days. The blood samples were taken by jugular venipuncture, after asepsis and stored at -20°C to its subsequent processing through Spectrophotometry. The results were tabulated in Excel and analyzed using statistical program SAS 9.1. The mean values found were: glucose 75.57 ± 27.5 mg/dL for females and 83.70 ± 37.7 mg/dL for males; total protein 8.47 ± 2.76 g/dL for females and 7.14 ± 3.26 g/dL for males; albumin 4.29 ± 1.41 g/dL for females and 3.83 ± 1.46 g/dL for males; globulin 4.62 ± 2.35 g/dL for females and 3.92 ± 2.45 g/dL for males. The day and the hour of sampling affect serum concentrations (p < 0.05 of evaluated metabolites, also there was variation of themself regarding sex (p < 0.05.

  6. Serum Total Homocysteine Level: A True Cardiovascular Risk Factor or an Acute Phase Reactant Protein?

    Directory of Open Access Journals (Sweden)

    Dr. Malik Asif Humayoun et al.

    2011-08-01

    Full Text Available Atherosclerosis is the leading cause of death in the developed world1.In Pakistan; the disease seems to follow an accelerated course with ischaemic events occurring a decade earlier than those reported worldwide2. While many risk factors of cardiovascular diseases are well established, many are still under evaluation; like homocysteine, which is now being considered an independent risk factor for cardiovascular diseases3.Homocysteine is a sulfur containing amino acid which is not our dietary constituent but is formed by metabolism of methionine, another amino acid present in our daily protein diet. In case of excess formation of homocysteine as compared to its consumption, it appears in urine if levels are too high3. Normal plasma homocysteine concentration ranges from 5 to 15μmol/litre. Of this, almost 75% is bound to proteins, especially albumin, through disulfide bond. Classification of hyperhomocysteinemia described by Kang SS 4 is followed widely.Moderate hyperhomocysteinemia (15 to 30 μmol/litreIntermediate hyperhomocysteinemia (30 to 100μmol/litreSevere hyperhomocysteinemia (>100 μmol/litre

  7. Elevated pre-treatment levels of plasma C-reactive protein are associated with poor prognosis after breast cancer

    DEFF Research Database (Denmark)

    Allin, Kristine H; Nordestgaard, Børge G; Flyger, Henrik;

    2011-01-01

    We examined whether plasma C-reactive protein (CRP) levels at the time of diagnosis of breast cancer are associated with overall survival, disease-free survival, death from breast cancer, and recurrence of breast cancer.......We examined whether plasma C-reactive protein (CRP) levels at the time of diagnosis of breast cancer are associated with overall survival, disease-free survival, death from breast cancer, and recurrence of breast cancer....

  8. Relation of resistin levels with C-reactve protein, homocysteine and uric acid in smokers and non-smokers

    OpenAIRE

    Onur Esbah; Gül Gürsoy; Nazli Gulsoy Kirnap; Hacer Cetiner; Berrin Demirbas; Yasar Acar; Murat Bayram

    2011-01-01

    Background: The association between C-reactive protein, homocysteine, uric acid levels and cardiovascular risk have been debated for decades. Resistin is a newly discovered adipocyte derived cytokine. Smoking besides its effect on atherosclerosis, is shown to alter adipocytokine levels. Bearing in mind, these complex relationship of resistin with smoking, C-reactive protein, homocysteine and uric acid, we planned to investigate the association of resistin and these cardiovascular risk factors...

  9. Signaling Pathways Related to Protein Synthesis and Amino Acid Concentration in Pig Skeletal Muscles Depend on the Dietary Protein Level, Genotype and Developmental Stages.

    Science.gov (United States)

    Liu, Yingying; Li, Fengna; Kong, Xiangfeng; Tan, Bie; Li, Yinghui; Duan, Yehui; Blachier, François; Hu, Chien-An A; Yin, Yulong

    2015-01-01

    Muscle growth is regulated by the homeostatic balance of the biosynthesis and degradation of muscle proteins. To elucidate the molecular interactions among diet, pig genotype, and physiological stage, we examined the effect of dietary protein concentration, pig genotype, and physiological stages on amino acid (AA) pools, protein deposition, and related signaling pathways in different types of skeletal muscles. The study used 48 Landrace pigs and 48 pure-bred Bama mini-pigs assigned to each of 2 dietary treatments: lower/GB (Chinese conventional diet)- or higher/NRC (National Research Council)-protein diet. Diets were fed from 5 weeks of age to respective market weights of each genotype. Samples of biceps femoris muscle (BFM, type I) and longissimus dorsi muscle (LDM, type II) were collected at nursery, growing, and finishing phases according to the physiological stage of each genotype, to determine the AA concentrations, mRNA levels for growth-related genes in muscles, and protein abundances of mechanistic target of rapamycin (mTOR) signaling pathway. Our data showed that the concentrations of most AAs in LDM and BFM of pigs increased (Ppigs had generally higher (Ppigs. The mRNA levels for myogenic determining factor, myogenin, myocyte-specific enhancer binding factor 2 A, and myostatin of Bama mini-pigs were higher (Ppigs, while total and phosphorylated protein levels for protein kinase B, mTOR, and p70 ribosomal protein S6 kinases (p70S6K), and ratios of p-mTOR/mTOR, p-AKT/AKT, and p-p70S6K/p70S6K were lower (Ppig genotype-dependent effect of dietary protein on the levels for mTOR and p70S6K. When compared with the higher protein-NRC diet, the lower protein-GB diet increased (Ppigs, but repressed (Ppigs. The higher protein-NRC diet increased ratio of p-mTOR/mTOR in Landrace pigs. These findings indicated that the dynamic consequences of AA profile and protein deposition in muscle tissues are the concerted effort of distinctive genotype, nutrient status, age, and

  10. Peningkatan Kadar Protein Putak melalui Fermentasi oleh Kapang Trichoderma reesei (THE INCREASE OF PROTEIN LEVEL FROM PUTAK THROUGH FERMENTATION OF FUNGI TRICHODERMA REESEI

    Directory of Open Access Journals (Sweden)

    Maritje Aleonor Hilakore

    2013-09-01

    Full Text Available A study was conducted was to increasing the protein level in putak by fermentation using fungiTrichoderma reesei. A laboratoryum experimental study was conducted using  factorial CompletelyRandomized Design 3 x 4 x 3. The main factor is were  inoculant levels of fungi T. reesei (T: 5,0; 7,5 and 10,0% (w/w,the level  and the second factor were of incubation time (W:  2; 3; and 4 days. Variables tested werecrude protein (CP, true protein (TP and crude fiber (CF. The result showed that  treatment with 7.5% ofT.reesei  and incubation time for 4 days gawe the highest of crude and true protein level (20,60%  from14,17% and 13,25% from 3,25%, and  lowest crude fiber 9,08% from 9,70%. Through fermentation of  fungiT.reesei can be increase the protein and decrease the fiber level of putak.

  11. Involvement of decreased neuroglobin protein level in cognitive dysfunction induced by 1-bromopropane in rats.

    Science.gov (United States)

    Guo, Ying; Yuan, Hua; Jiang, Lulu; Yang, Junlin; Zeng, Tao; Xie, Keqin; Zhang, Cuili; Zhao, Xiulan

    2015-03-10

    1-Bromopropane (1-BP) is used as a substitute for ozone-depleting solvents (ODS) in industrial applications. 1-BP could display central nervous system (CNS) neurotoxicity manifested by cognitive dysfunction. Neuroglobin (Ngb) is an endogenous neuroprotectant and is predominantly expressed in the nervous system. The present study aimed to investigate Ngb involvement in CNS neurotoxicity induced by 1-BP in rats. Male Wistar rats were randomly divided into 5 groups (n=14) and treated with 0, 100, 200, 400 and 800 mg/kg bw 1-BP, respectively, by gavage for consecutive 12 days. Rats displayed cognitive dysfunction dose-dependently through Morris water maze (MWM) test. Significant neuron loss in layer 5 of the prelimbic cortex (PL) was observed. Moreover, 1-BP decreased Ngb protein level in cerebral cortex and Ngb decrease was significantly positively correlated with cognitive dysfunction. Glutathione (GSH) content, GSH/oxidized glutathione (GSSG) ratio and glutamate cysteine ligase (GCL) activity decreased in cerebral cortex, coupled with the increase in GSSG content. GSH and GSH/GSSG ratio decrease were significantly positively correlated with cortical Ngb decrease. Additionally, levels of N-epsilon-hexanoyl-lysine (HEL) and 4-hydroxy-2-nonenal (4-HNE) modified proteins in cerebral cortex of 1-BP-treated rats increased significantly. In conclusion, it was suggested that 1-BP resulted in decreased endogenous neuroprotectant Ngb in cerebral cortex, which might play an important role in CNS neurotoxicity induced by 1-BP and that 1-BP-induced oxidative stress in cerebral cortex might partly be responsible for Ngb decrease.

  12. Blood profiling of proteins and steroids during weight maintenance with manipulation of dietary protein level and glycaemic index

    DEFF Research Database (Denmark)

    Wang, Ping; Holst, Claus; Astrup, Arne

    2012-01-01

    Weight regain after weight loss is common. In the Diogenes dietary intervention study, a high-protein and low-glycaemic index (GI) diet improved weight maintenance. The objective of the present study was to identify (1) blood profiles associated with continued weight loss and weight regain (2) bl...... differences between continued weight losers and weight regainers. Increases in leptin (LEP) and C-reactive protein (CRP) were significantly associated with weight regain (P ...

  13. Levels of common salivary protein 1 in healthy subjects and periodontal patients

    Science.gov (United States)

    2016-01-01

    Purpose Human saliva, as a vital part of the immune defense system, contains a number of distinct proteins and peptides. Recently human common salivary protein 1 (CSP1) has been identified as an abundant salivary protein and may play a role in promoting the binding of cariogenic bacteria to salivary pellicles. However, nothing else is known regarding the role of CSP1 in periodontology. The aim of this study was to quantify and compare CSP1 levels between healthy subjects and periodontal patients. Methods This controlled clinical study was conducted in periodontally healthy individuals and patients with chronic periodontitis Chonbuk National University Hospital, with Institutional Review Board approval. Whole saliva samples were collected from 36 healthy subjects and 33 chronic periodontitis patients and analyzed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immune blotting were conducted to ensure that anti-CSP1 monoclonal antibody (mAb) binds to CSP1 in human saliva. A sandwich enzyme-linked immunosorbent assay (ELISA) system was house-fabricated using mAb-hCSP1#14 and mAb-hCSP1#4 as a capture and a detector mAb, respectively. The CSP1 concentrations in saliva from 36 healthy subjects and 33 periodontal patients were quantified using the CSP1 sandwich ELISA system, and the results were analyzed using the Student’s t-test. Results Immunoblot analysis using mAb-hCSP1 as a probe confirmed that CSP1 in human saliva existed as a single band with a molecular weight of approximately 27-kDa. The quantification of CSP1 concentrations by CSP1 ELISA showed that the median values (25th to 75th percentiles) of periodontal patients and healthy subjects were 9,474 ng/mL (range, 8,434–10,139 ng/mL) and 8,598 ng/mL (range, 7,421–9,877 ng/mL), respectively. The Student’s t-test indicated the presence of a statistically significant difference between the 2 groups (P=0.024). Conclusions The presence of a significant difference in CSP1 levels between healthy

  14. Carotenoid crystal formation in Arabidopsis and carrot roots caused by increased phytoene synthase protein levels.

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    Dirk Maass

    Full Text Available BACKGROUND: As the first pathway-specific enzyme in carotenoid biosynthesis, phytoene synthase (PSY is a prime regulatory target. This includes a number of biotechnological approaches that have successfully increased the carotenoid content in agronomically relevant non-green plant tissues through tissue-specific PSY overexpression. We investigated the differential effects of constitutive AtPSY overexpression in green and non-green cells of transgenic Arabidopsis lines. This revealed striking similarities to the situation found in orange carrot roots with respect to carotenoid amounts and sequestration mechanism. METHODOLOGY/PRINCIPAL FINDINGS: In Arabidopsis seedlings, carotenoid content remained unaffected by increased AtPSY levels although the protein was almost quantitatively imported into plastids, as shown by western blot analyses. In contrast, non-photosynthetic calli and roots overexpressing AtPSY accumulated carotenoids 10 and 100-fold above the corresponding wild-type tissues and contained 1800 and 500 microg carotenoids per g dry weight, respectively. This increase coincided with a change of the pattern of accumulated carotenoids, as xanthophylls decreased relative to beta-carotene and carotene intermediates accumulated. As shown by polarization microscopy, carotenoids were found deposited in crystals, similar to crystalline-type chromoplasts of non-green tissues present in several other taxa. In fact, orange-colored carrots showed a similar situation with increased PSY protein as well as carotenoid levels and accumulation patterns whereas wild white-rooted carrots were similar to Arabidopsis wild type roots in this respect. Initiation of carotenoid crystal formation by increased PSY protein amounts was further confirmed by overexpressing crtB, a bacterial PSY gene, in white carrots, resulting in increased carotenoid amounts deposited in crystals. CONCLUSIONS: The sequestration of carotenoids into crystals can be driven by the

  15. Serum high-sensitivity C-reaction protein and heart fatty acid binding protein level and cardiac accidents in patients with unstable angina pectoris

    Institute of Scientific and Technical Information of China (English)

    朱红秋

    2006-01-01

    Objective To investigate the relationship between serum high-sensitivity C-reaction protein (hs-CRP) and heart fatty acid binding protein (h-FABP) on cardiac accidents in patients with unstable angina pectoris (UAP). Methods Serum levels of hs-CRP, h-FABP, cardiac troponin-Ⅰ(cTn-Ⅰ) and creatine kinase MB isoenzyme (CK-MB) were measured and cardiac accidents within 2 weeks after the test were observed in 74 patients (male

  16. High-level expression, purification and characterization of codon-optimized recombinant hemagglutinin 5 proteins in mammalian cells

    Institute of Scientific and Technical Information of China (English)

    YANG Jing-lin; WANG Hong-liang; WANG Shun-xin; YANG Peng; LIU Kang-tai; JIANG Cheng-yu

    2010-01-01

    Background Numerous Asian cases of avian influenza virus infection, especially the highly pathogenic strain H5N1, in humans have raised the concern that another influenza pandemic is close. However, there are no effective therapeutic drugs or preventative vaccines available. Hemagglutinin is the membrane glycoprotein of avian influenza virus responsible for receptor binding to human cells and the main immunogenic protein that elicits a strong immune response. Although this protein is of great importance to the study of pathogenesis and vaccine development, its expression and purification are difficult due to high levels of glycosylation.Methods In this study, we expressed codon-optimized, full-length hemagglutinin 5 (H5) protein fused with a human IgG Fc tag (H5-Fc) in HEK293 cells. To enhance secretion of this protein, we also deleted the transmembrane domain and the intracellular domain of the H5 protein (H5ΔTM-Fc). Purified proteins were obtained using a protein A column. Results ELISA revealed that the yield of soluble H5ATM-Fc protein in the supernatant was about 20 mg/L. Western blotting and fluorescence activated cell sorter (FACS) indicated that the purified H5 protein was correctly folded and biologically active.Conclusion Purification of H5 proteins from mammalian cells could be used for large-scale production of recombinant H5 protein for basic scientific research or the development of vaccines.

  17. Discrepant NOXA (PMAIP1) transcript and NOXA protein levels: a potential Achilles' heel in mantle cell lymphoma

    OpenAIRE

    2014-01-01

    Mantle cell lymphoma (MCL) is an aggressive lymphoid neoplasm with transient response to conventional chemotherapy. We here investigated the role of the Bcl-2 homology domain 3-only protein NOXA for life–death decision in MCL. Surprisingly, NOXA (PMAIP1) mRNA and NOXA protein levels were extremely discrepant in MCL cells: NOXA mRNA was found to be highly expressed whereas NOXA protein levels were low. Chronic active B-cell receptor signaling and to a minor degree cyclin D1 overexpression cont...

  18. Decreased dietary protein or energy intake and plasma growth hormone levels of the pregnant pig, its fetuses and developing progeny.

    Science.gov (United States)

    Atinmo, T; Baldijrao, C; Pond, W G; Barnes, R H

    1976-07-01

    The effects of low protein diets on plasma growth hormone were studied in pregnant pigs, fetuses and the developing progeny. Pregnant pigs were fed 18%, 3% or 0.5% protein diet throughout the gestation period. At 10, 13 and 15 week of gestation, fetuses were removed from the uterus after the dam had been bled to death. Plasma samples were used for growth hormone determinations. In a second experiment, 2-day old pigs from another set of pregnant pigs fed the diet containing 18%, 3% or 0.5% protein during gestation were cross-fostered to control nursing dams and weaned at 4 weeks of age to a standard diet. Plasma obtained at regular intervals was used for growth hormone determination. Plasma growth hormone was significantly higher in dams fed 0.5% protein after week 13 of gestation. High growth hormone (ten times the dam GH level) was observed in all fetuses irrespective of maternal dietary manipulation. Offspring of severely protein deprived pits (0.5% protein) had significantly elevated growth hormone levels up to 12 weeks of age in spite of cross fostering to a control dam after birth. The data suggest that there is little or no effect of maternal protein restriction on fetal growth hormone levels but the persistent high growth hormone levels in the progeny of severely malnourished pigs indicate a possible impairment of the production, release or catabolism of growth hormone and/or its releasing factor.

  19. Mass-tag labeling reveals site-specific and endogenous levels of protein S-fatty acylation.

    Science.gov (United States)

    Percher, Avital; Ramakrishnan, Srinivasan; Thinon, Emmanuelle; Yuan, Xiaoqiu; Yount, Jacob S; Hang, Howard C

    2016-04-19

    Fatty acylation of cysteine residues provides spatial and temporal control of protein function in cells and regulates important biological pathways in eukaryotes. Although recent methods have improved the detection and proteomic analysis of cysteine fatty (S-fatty) acylated proteins, understanding how specific sites and quantitative levels of this posttranslational modification modulate cellular pathways are still challenging. To analyze the endogenous levels of protein S-fatty acylation in cells, we developed a mass-tag labeling method based on hydroxylamine-sensitivity of thioesters and selective maleimide-modification of cysteines, termed acyl-PEG exchange (APE). We demonstrate that APE enables sensitive detection of protein S-acylation levels and is broadly applicable to different classes of S-palmitoylated membrane proteins. Using APE, we show that endogenous interferon-induced transmembrane protein 3 is S-fatty acylated on three cysteine residues and site-specific modification of highly conserved cysteines are crucial for the antiviral activity of this IFN-stimulated immune effector. APE therefore provides a general and sensitive method for analyzing the endogenous levels of protein S-fatty acylation and should facilitate quantitative studies of this regulated and dynamic lipid modification in biological systems.

  20. Cell cycle regulation of tubulin RNA level, tubulin protein synthesis, and assembly of microtubules in Physarum.

    Science.gov (United States)

    Schedl, T; Burland, T G; Gull, K; Dove, W F

    1984-07-01

    The temporal relationship between tubulin expression and the assembly of the mitotic spindle microtubules has been investigated during the naturally synchronous cell cycle of the Physarum plasmodium. The cell cycle behavior of the tubulin isoforms was examined by two-dimensional gel electrophoresis of proteins labeled in vivo and by translation of RNA in vitro. alpha 1-, alpha 2-, beta 1-, and beta 2-tubulin synthesis increases coordinately until metaphase, and then falls, with beta 2 falling more rapidly than beta 1. Nucleic acid hybridization demonstrated that alpha- and beta-tubulin RNAs accumulate coordinately during G2, peaking at metaphase. Quantitative analysis demonstrated that alpha-tubulin RNA increases with apparent exponential kinetics, peaking with an increase over the basal level of greater than 40-fold. After metaphase, tubulin RNA levels fall exponentially, with a short half-life (19 min). Electron microscopic analysis of the plasmodium showed that the accumulation of tubulin RNA begins long before the polymerization of mitotic spindle microtubules. By contrast, the decay of tubulin RNA after metaphase coincides with the depolymerization of the spindle microtubules.

  1. Immunological Roles of Elevated Plasma Levels of Matricellular Proteins in Japanese Patients with Pulmonary Tuberculosis

    Directory of Open Access Journals (Sweden)

    Beata Shiratori

    2016-12-01

    Full Text Available Elevated matricellular proteins (MCPs, including osteopontin (OPN and galectin-9 (Gal-9, were observed in the plasma of patients with Manila-type tuberculosis (TB previously. Here, we quantified plasma OPN, Gal-9, and soluble CD44 (sCD44 by enzyme-linked immunosorbent assay (ELISA, and another 29 cytokines by Luminex assay in 36 patients with pulmonary TB, six subjects with latent tuberculosis (LTBI, and 19 healthy controls (HCs from Japan for a better understanding of the roles of MCPs in TB. All TB subjects showed positive results of enzyme-linked immunospot assays (ELISPOTs. Spoligotyping showed that 20 out of 36 Mycobacterium tuberculosis (MTB strains belong to the Beijing type. The levels of OPN, Gal-9, and sCD44 were higher in TB (positivity of 61.1%, 66.7%, and 63.9%, respectively than in the HCs. Positive correlations between OPN and Gal-9, between OPN and sCD44, and negative correlation between OPN and ESAT-6-ELISPOT response, between chest X-ray severity score of cavitary TB and ESAT-6-ELISPOT response were observed. Instead of OPN, Gal-9, and sCD44, cytokines G-CSF, GM-CSF, IFN-α, IFN-γ, IL-12p70, and IL-1RA levels were higher in Beijing MTB-infected patients. These findings suggest immunoregulatory, rather than inflammatory, effect of MCPs and can advance the understanding of the roles of MCPs in the context of TB pathology.

  2. Protein pheromone expression levels predict and respond to the formation of social dominance networks.

    Science.gov (United States)

    Nelson, A C; Cunningham, C B; Ruff, J S; Potts, W K

    2015-06-01

    Communication signals are key regulators of social networks and are thought to be under selective pressure to honestly reflect social status, including dominance status. The odours of dominants and nondominants differentially influence behaviour, and identification of the specific pheromones associated with, and predictive of, dominance status is essential for understanding the mechanisms of network formation and maintenance. In mice, major urinary proteins (MUPs) are excreted in extraordinary large quantities and expression level has been hypothesized to provide an honest signal of dominance status. Here, we evaluate whether MUPs are associated with dominance in wild-derived mice by analysing expression levels before, during and after competition for reproductive resources over 3 days. During competition, dominant males have 24% greater urinary MUP expression than nondominants. The MUP darcin, a pheromone that stimulates female attraction, is predictive of dominance status: dominant males have higher darcin expression before competition. Dominants also have a higher ratio of darcin to other MUPs before and during competition. These differences appear transient, because there are no differences in MUPs or darcin after competition. We also find MUP expression is affected by sire dominance status: socially naive sons of dominant males have lower MUP expression, but this apparent repression is released during competition. A requisite condition for the evolution of communication signals is honesty, and we provide novel insight into pheromones and social networks by showing that MUP and darcin expression is a reliable signal of dominance status, a primary determinant of male fitness in many species.

  3. Influence of morphine on levels of type Ⅱ inhibitory guanine nucleotide binding protein in primary hippocampal neurons

    Institute of Scientific and Technical Information of China (English)

    Qinghua Wu; Qiang Fu; Xinhua Wang; Jianhua Zhao; Liwei Liu; Shirong Tang

    2008-01-01

    BACKGROUND: The pharmacological action of opioid drugs is related to signal transduction of inhibitory guanine nucleotide binding protein.OBJECTIVE: To quantitatively and qualitatively analyze the influence of morphine on levels of type Ⅱ inhibitory guanine nucleotide binding protein (Gi2 protein) in primary cultured hippocampal neurons at different time points.DESIGN, TIME AND SETTING: A randomized controlled study, which was performed at the Department of Neurobiology, Changzheng Hospital, Second Military Medical University of Chinese PLA between September 2002 and March 2004.MATERIALS: Cerebral hippocampal neurons were obtained from newborn SD rats at 1-2 days of age. Biotin-antibody Ⅱ-avidin fluorescein isothiocyanate (Avidin-FITC) was purchased from Sigma Company (USA) and the Gi2 protein polyclonal antibody from Santa Cruz Biochemistry Company (USA).METHODS: Seven days after culture, mature hippocampal neurons were randomly divided into six groups: 4-, 8-, 16-, 24-, and 48-hour morphine groups, and a blank control group. Neurons in the morphine groups Received morphine (10μmol/L), which could cause alterations of G-protein mRNA and cAMP expression in the prefrontal cortex. Neurons in the blank control group were given the same volume of saline.MAIN OUTCOME MEASURES: Gi2 protein levels were detected by an immunofluorescence technique, and were analyzed by the image analytic system with the use of green fluorescence intensity.RESULTS: Gi2 protein levels in hippocampal neurons gradually decreased in the 4-, 8-, 16-, 24-, and 48-hour morphine groups. In particular, Gi2 protein levels in the 16-, 24-, and 48-hour morphine groups were significantly lower than that in the blank control group (P<0.05-0.01).CONCLUSION: Morphine may decrease Gi2 protein level in primary hippocampal neurons, and the decreasing trend is positively related to morphine-induced time.

  4. Adrenal G protein-coupled receptor kinase-2 in regulation of sympathetic nervous system activity in heart failure

    Institute of Scientific and Technical Information of China (English)

    Katie; A; Mc; Crink; Ava; Brill; Anastasios; Lymperopoulos

    2015-01-01

    Heart failure(HF), the number one cause of death in the western world, is caused by the insufficient performance of the heart leading to tissue underperfusion in response to an injury or insult. It comprises complex interactions between important neurohormonal mechanisms that try but ultimately fail to sustain cardiac output. The most prominent such mechanism is the sympathetic(adrenergic) nervous system(SNS), whose activity and outflow are greatly elevated in HF. SNS hyperactivity confers significant toxicity to the failing heart and markedly increases HF morbidity and mortality via excessive activation of adrenergic receptors, which are G protein-coupled receptors. Thus, ligand binding induces their coupling to heterotrimeric G proteins that transduce intracellular signals. G protein signaling is turned-off by the agonist-bound receptor phosphorylation courtesy of G protein-coupled receptor kinases(GRKs), followed by βarrestin binding, which prevents the GRK-phosphorylated receptor from further interaction with the G proteins and simultaneously leads it inside the cell(receptor sequestration). Recent evidence indicates that adrenal GRK2 and βarrestins can regulate adrenal catecholamine secretion, thereby modulating SNS activity in HF. The present review gives an account of all these studies on adrenal GRKs and βarrestins in HF and discusses the exciting new therapeutic possibilities for chronic HF offered by targeting these proteins pharmacologically.

  5. Inhibition of SIRT1 Increases EZH2 Protein Level and Enhances the Repression of EZH2 on Target Gene Expression

    Institute of Scientific and Technical Information of China (English)

    Lu Lu; Lei Li; Xiang Lü; Xue-song Wu; De-pei Liu; Chih-chuan Liang

    2011-01-01

    Objective To study the regulatory roles of SIRT1 on EZH2 expression and the further effects on EZH2's repression of target gene expression. Methods The stable SIRT1 RNAi and Control RNAi HeLa cells were established by infection with retroviruses expressing shSIRT1 and shLuc respectively followed by puromycin selection. EZH2 protein level was detected by Western blot in either whole cell lysate or the fractional cell extract. Reverse transcription-polymerase chain reaction was performed to detect the mRNA level of EZH2. Cycloheximide was used to treat SIRT1 RNAi and Control RNAi cells for protein stability assay. Chromatin immunoprecipitation (CHIP) assay was applied to measure enrichment of SIRT1, EZH2, and trimethylated H3K27 (H3K27me3) at SATB1 promoter in SIRT1 RNAi and Control RNAi cells.Results Western blot results showed that EZH2 protein level increased upon SIRT1 depletion. Fractional extraction results showed unchanged cytoplasmic fraction and increased chromatin fraction of EZH2 protein in SIRTI RNAi cells. The mRNA level of EZH2 was not affected by knockdown of SIRT1. SIRT1 recruitment was not detected at the promoter region of EZH2 gene locus. The protein stability assay showed that the protein stability of EZH2 increases upon SIRTI knockdown. Upon SIRT1 depletion, EZH2 and H3K27me3 recruitment at SATB1 promoter increases and the mRNA level of SATB1 decreases.Conclusions Depletion of SIRT1 increases the protein stability of EZH2. The regulation of EZH2 protein level by SIRTI affects the repressive effects of EZH2 on the target gene expression.

  6. Comparison of serum placental protein hormone levels in diabetic and normal pregnancy.

    Science.gov (United States)

    Braunstein, G D; Mills, J L; Reed, G F; Jovanovic, L G; Holmes, L B; Aarons, J; Simpson, J L

    1989-01-01

    Conflicting data exist concerning maternal serum concentrations of placental hormones during pregnancy in women with diabetes mellitus. To resolve some of these discrepancies, women participating in the NICHD-Diabetes in Early Pregnancy Study were studied. In this collaborative study, pregnancy was identified within 21 days of conception by serum hCG measurements. We prospectively collected 185 blood samples from 35 insulin-dependent diabetic women and 166 blood samples from 31 control women, all between 5 and 37 weeks gestation. Serum concentrations of hCG, pregnancy-specific beta-1-glycoprotein, placental lactogen, and hCG alpha were measured serially. The relationship between serum hormone, fasting blood glucose, 1-h postprandial blood glucose, and glycosylated hemoglobin concentrations was compared. Serum hCG alpha levels were significantly lower in the diabetic women than in control women at multiple time points during the first and second trimesters, while no consistent differences in the serum concentrations of hCG or pregnancy-specific beta-1-glycoprotein were found between pregnant diabetic and control women. Serum placental lactogen levels were significantly lower in diabetic women at 9-10 weeks and 20 weeks gestation. There were no correlations between fasting blood glucose, 1-h postprandial blood glucose, or glycosylated hemoglobin and any of the placental protein levels in the diabetic women. These data are consistent with a defect in synthesis and/or secretion of hCG alpha by the cytotrophoblast during the first two trimesters of pregnancy in insulin-requiring diabetic women.

  7. Blood levels of glial fibrillary acidic protein (GFAP in patients with neurological diseases.

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    Christoph A Mayer

    Full Text Available BACKGROUND AND PURPOSE: The brain-specific astroglial protein GFAP is a blood biomarker candidate indicative of intracerebral hemorrhage in patients with symptoms suspicious of acute stroke. Comparably little, however, is known about GFAP release in other neurological disorders. In order to identify potential "specificity gaps" of a future GFAP test used to diagnose intracerebral hemorrhage, we measured GFAP in the blood of a large and rather unselected collective of patients with neurological diseases. METHODS: Within a one-year period, we randomly selected in-patients of our university hospital for study inclusion. Patients with ischemic stroke, transient ischemic attack and intracerebral hemorrhage were excluded. Primary endpoint was the ICD-10 coded diagnosis reached at discharge. During hospital stay, blood was collected, and GFAP plasma levels were determined using an advanced prototype immunoassay at Roche Diagnostics. RESULTS: A total of 331 patients were included, covering a broad spectrum of neurological diseases. GFAP levels were low in the vast majority of patients, with 98.5% of cases lying below the cut-off that was previously defined for the differentiation of intracerebral hemorrhage and ischemic stroke. No diagnosis or group of diagnoses was identified that showed consistently increased GFAP values. No association with age and sex was found. CONCLUSION: Most acute and chronic neurological diseases, including typical stroke mimics, are not associated with detectable GFAP levels in the bloodstream. Our findings underline the hypothesis that rapid astroglial destruction as in acute intracerebral hemorrhage is mandatory for GFAP increase. A future GFAP blood test applied to identify patients with intracerebral hemorrhage is likely to have a high specificity.

  8. 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 ...

  9. Effect of Lower Protein Level in Feed on Production Performance of Zagorje Turkey

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    Stjepan Mužić

    2003-06-01

    Full Text Available Interest for production and marketing of Zagorje turkey in Croatia is rising. Traditional way of breeding Zagorje turkey is manifested through the variety of production and nutritional practices. Typical Croatian customer demands a carcass of Zagorje turkey that has a low weight, ranging between 2.5 . 3.5 kg and is pre-processed in the slaughterhouse. As the data on nutrition, breeding and production of light autochthonous turkey breeds, particularly, among them, Zagorje turkey, are insufficient; there is a need for research on the nutritional and technological aspects of free-range production of Zagorje turkey. In that direction we carried out the research of the effect of different protein level in the diet of turkey chicks up to 8 weeks of age to the final production performance and meat quality of Zagorje turkey slaughtered at the age of six months. Two control (68 ♂ and 68 ♀ and two experimental groups (68 ♂ and 68 ♀ of Zagorje turkeys were housed indoors for the first 8 weeks. After that period each group had access to a fenced pasture of 0.5 ha. Both experimental groups were fed a diet with 25 % less protein than the control groups during the first period of 8 weeks, while after this period all of the groups were fed equally until the end of month 6. At the end of production period, birds of both sexes of the experimental groups had a lower body weight that the control ones, but still within the suitable category for Croatian consumers, while males from the control groups were too heavy (6854 g. Feed conversion ratio (FCR over the whole experimental period both in the control and experimental groups ranged between 3.47-4.06 kg (concentrates only, without a pattern considering treatment or sex. In the same time, share of different parts of body in the carcass differed between treatments and sexes.

  10. Krill protein hydrolysate reduces plasma triacylglycerol level with concurrent increase in plasma bile acid level and hepatic fatty acid catabolism in high-fat fed mice

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    Marie S. Ramsvik

    2013-11-01

    Full Text Available Background: Krill powder, consisting of both lipids and proteins, has been reported to modulate hepatic lipid catabolism in animals. Fish protein hydrolysate diets have also been reported to affect lipid metabolism and to elevate bile acid (BA level in plasma. BA interacts with a number of nuclear receptors and thus affects a variety of signaling pathways, including very low density lipoprotein (VLDL secretion. The aim of the present study was to investigate whether a krill protein hydrolysate (KPH could affect lipid and BA metabolism in mice. Method: C57BL/6 mice were fed a high-fat (21%, w/w diet containing 20% crude protein (w/w as casein (control group or KPH for 6 weeks. Lipids and fatty acid composition were measured from plasma, enzyme activity and gene expression were analyzed from liver samples, and BA was measured from plasma. Results: The effect of dietary treatment with KPH resulted in reduced levels of plasma triacylglycerols (TAG and non-esterified fatty acids (NEFAs. The KPH treated mice had also a marked increased plasma BA concentration. The increased plasma BA level was associated with induction of genes related to membrane canalicular exporter proteins (Abcc2, Abcb4 and to BA exporters to blood (Abcc3 and Abcc4. Of note, we observed a 2-fold increased nuclear farnesoid X receptor (Fxr mRNA levels in the liver of mice fed KPH. We also observed increased activity of the nuclear peroxiosme proliferator-activated receptor alpha (PPARα target gene carnitine plamitoyltransferase 2 (CPT-2. Conclusion: The KPH diet showed to influence lipid and BA metabolism in high-fat fed mice. Moreover, increased mitochondrial fatty acid oxidation and elevation of BA concentration may regulate the plasma level of TAGs and NEFAs.

  11. Evidence that kidney function but not type 2 diabetes determines retinol-binding protein 4 serum levels

    DEFF Research Database (Denmark)

    Henze, Andrea; Frey, Simone K; Raila, Jens

    2008-01-01

    It has been suggested that retinol-binding protein 4 (RBP4) links adiposity, insulin resistance, and type 2 diabetes. However, circulating RBP4 levels are also affected by kidney function. Therefore, the aim of this study was to test whether RBP4 serum levels are primarily associated with kidney...

  12. Pregnancy-associated plasma protein A in human ovarian follicles and its association with intrafollicular hormone levels

    DEFF Research Database (Denmark)

    Bøtkjær, Jane Alrø; Jeppesen, Janni Vikkelsø; Wissing, Marie Louise

    2015-01-01

    To evaluate follicular fluid (FF) levels of pregnancy-associated plasma protein A (PAPP-A) in relation to levels of intrafollicular hormones. Furthermore, immunostaining of human follicles of varying diameters was studied for PAPP-A, antimüllerian hormone (AMH), and aromatase, and the biological ...

  13. A Pelargonium ARGONAUTE4 gene shows organ-specific expression and differences in RNA and protein levels.

    Science.gov (United States)

    He, Jie; Gray, John; Leisner, Scott

    2010-03-01

    RNAi-induced gene silencing plays a role in plant DNA methylation and defense. While most gene silencing studies have been performed on annuals, little is known about the expression of key components of this process (like ARGONAUTE proteins) in ornamentals. Using a combination of polymerase chain reaction techniques, an ARGONAUTE4 gene, PhAGO4, was isolated from Pelargonium. PhAGO4 encodes a predicted product of 934 amino acids that contains the PAZ and PIWI domains typical of ARGONAUTE (AGO) proteins. Phylogenetic analyses indicate that PhAGO4 clusters with other plant AGO4 proteins. Organ expression patterns of the AGO4 genes in Pelargonium and Arabidopsis show intriguing differences. AGO4 RNA levels decline with leaf age in both Arabidopsis and Pelargonium. In contrast AGO4 RNA levels in roots relative to leaves are higher in Pelargonium than in Arabidopsis. Both Arabidopsis and Pelargonium AGO4 showed higher RNA levels in flowers than leaves or roots. Even though flowers show higher levels of PhAGO4 RNA when compared to leaves and roots, protein gel blot analysis shows that at the protein level, the reverse is true. This suggests that PhAGO4 expression may be regulated at the translational or post-translational level in Pelargonium flowers.

  14. [Relationship between the included levels of coffee pulp and the protein content in rations for monogastric animals].

    Science.gov (United States)

    Gómez-Brenes, R A; Bendaña, G; González, J M; Braham, J E; Bressani, R

    1985-09-01

    The purpose of this research was to determine the effect of including fresh and ensilaged coffee pulp in rations for monogastric animals, and find the best protein and coffee pulp levels in rations for rats. Fresh coffee pulp and pulp ensilaged for 12 months were used; both kinds of pulp were sun-dried before incorporating them into the rations. The chemical analyses of the pulps revealed a lower content in caffeine, tannins, chlorogenic acid and caffeic acid in the ensilaged pulp than in fresh coffee pulp. Thirty-two experimental rations were prepared, 16 with fresh coffee pulp and 16 with the ensilaged by-product, distributed into four different protein levels (10, 15, 20 and 25%), and three levels of pulp (15, 30 and 45%) for each protein level. The rations thus prepared were fed to Wistar albino rats for a six-week period. The parameters used to measure the effect of the two types of pulp were mortality rate, food consumption, weight gain, food conversion and apparent digestibility of the rations. Ensilaged pulp had a higher nutritive value, lower toxicity and better digestibility than fresh pulp. The increase in the protein level of the ration resulted in partial protection against the negative effects of coffee pulp on the performance of animals, since this improved as the protein level of the ration increased.

  15. Ileal Mucosal and Fecal Pancreatitis Associated Protein Levels Reflect Severity of Salmonella Inflection in Rats

    NARCIS (Netherlands)

    Ampting, van M.T.J.; Rodenburg, G.C.H.; Vink, C.; Kramer, E.; Schonewille, A.; Keijer, J.; Meer, van der R.; Bovee-Oudenhoven, I.M.J.

    2009-01-01

    Background Microbial infections induce ileal pancreatitis-associated protein/regenerating gene III (PAP/RegIII) mRNA expression. Despite increasing interest, little is known about the PAP/RegIII protein. Therefore, ileal mucosal PAP/RegIII protein expression, localization, and fecal excretion were s

  16. Parkinson's disease-associated kinase PINK1 regulates Miro protein level and axonal transport of mitochondria.

    Directory of Open Access Journals (Sweden)

    Song Liu

    Full Text Available Mutations in Pten-induced kinase 1 (PINK1 are linked to early-onset familial Parkinson's disease (FPD. PINK1 has previously been implicated in mitochondrial fission/fusion dynamics, quality control, and electron transport chain function. However, it is not clear how these processes are interconnected and whether they are sufficient to explain all aspects of PINK1 pathogenesis. Here we show that PINK1 also controls mitochondrial motility. In Drosophila, downregulation of dMiro or other components of the mitochondrial transport machinery rescued dPINK1 mutant phenotypes in the muscle and dopaminergic (DA neurons, whereas dMiro overexpression alone caused DA neuron loss. dMiro protein level was increased in dPINK1 mutant but decreased in dPINK1 or dParkin overexpression conditions. In Drosophila larval motor neurons, overexpression of dPINK1 inhibited axonal mitochondria transport in both anterograde and retrograde directions, whereas dPINK1 knockdown promoted anterograde transport. In HeLa cells, overexpressed hPINK1 worked together with hParkin, another FPD gene, to regulate the ubiquitination and degradation of hMiro1 and hMiro2, apparently in a Ser-156 phosphorylation-independent manner. Also in HeLa cells, loss of hMiro promoted the perinuclear clustering of mitochondria and facilitated autophagy of damaged mitochondria, effects previously associated with activation of the PINK1/Parkin pathway. These newly identified functions of PINK1/Parkin and Miro in mitochondrial transport and mitophagy contribute to our understanding of the complex interplays in mitochondrial quality control that are critically involved in PD pathogenesis, and they may explain the peripheral neuropathy symptoms seen in some PD patients carrying particular PINK1 or Parkin mutations. Moreover, the different effects of loss of PINK1 function on Miro protein level in Drosophila and mouse cells may offer one explanation of the distinct phenotypic manifestations of PINK1

  17. Determining synthesis rates of individual proteins in zebrafish (Danio rerio) with low levels of a stable isotope labelled amino acid.

    Science.gov (United States)

    Geary, Bethany; Magee, Kieran; Cash, Phillip; Young, Iain S; Whitfield, Phillip D; Doherty, Mary K

    2016-05-01

    The zebrafish is a powerful model organism for the analysis of human cardiovascular development and disease. Understanding these processes at the protein level not only requires changes in protein concentration to be determined but also the rate at which these changes occur on a protein-by-protein basis. The ability to measure protein synthesis and degradation rates on a proteome-wide scale, using stable isotope labelling in conjunction with mass spectrometry is now a well-established experimental approach. With the advent of more selective and sensitive mass spectrometers, it is possible to accurately measure lower levels of stable isotope incorporation, even when sample is limited. In order to challenge the sensitivity of this approach, we successfully determined the synthesis rates of over 600 proteins from the cardiac muscle of the zebrafish using a diet where either 30% or 50% of the L-leucine was replaced with a stable isotope labelled analogue ([(2) H7 ]L-leucine]. It was possible to extract sufficient protein from individual zebrafish hearts to determine the incorporation rate of the label into hundreds of proteins simultaneously, with the two labelling regimens showing a good correlation of synthesis rates.

  18. Protein debradability and transformation of miimosine from leucaena Leucocephala (LAM de with in the rumen of buffaloes under diets with different protein levels

    Directory of Open Access Journals (Sweden)

    Raul Franzolin Neto

    1992-12-01

    Full Text Available The ruminal suspended nylon bags technique was used to study the degradability of crude protein and disintegration of minosin of Leucaena leucocephala in water buffaloes. The experimental desing was in a 4 x 4 Latin square, with variable levels of crude protein (8.8, 10,5, 12,5, and 14.9% of dry matter. The diets did not cause significant differences on the ruminal degradation of crude protein of Leucaena (P > 0.01, that ranged from 30.92 to 71.70% in, respectively, 3 or 48 hours of ruminal exposition. There was observed a clow ruminal protein degradation and a strong desingration of mimosin on the samples of Leucaena exposed in nylon bags to the rumen of water buffaloes

  19. Atomic-level description of protein-lipid interactions using an accelerated membrane model.

    Science.gov (United States)

    Baylon, Javier L; Vermaas, Josh V; Muller, Melanie P; Arcario, Mark J; Pogorelov, Taras V; Tajkhorshid, Emad

    2016-07-01

    Peripheral membrane proteins are structurally diverse proteins that are involved in fundamental cellular processes. Their activity of these proteins is frequently modulated through their interaction with cellular membranes, and as a result techniques to study the interfacial interaction between peripheral proteins and the membrane are in high demand. Due to the fluid nature of the membrane and the reversibility of protein-membrane interactions, the experimental study of these systems remains a challenging task. Molecular dynamics simulations offer a suitable approach to study protein-lipid interactions; however, the slow dynamics of the lipids often prevents sufficient sampling of specific membrane-protein interactions in atomistic simulations. To increase lipid dynamics while preserving the atomistic detail of protein-lipid interactions, in the highly mobile membrane-mimetic (HMMM) model the membrane core is replaced by an organic solvent, while short-tailed lipids provide a nearly complete representation of natural lipids at the organic solvent/water interface. Here, we present a brief introduction and a summary of recent applications of the HMMM to study different membrane proteins, complementing the experimental characterization of the presented systems, and we offer a perspective of future applications of the HMMM to study other classes of membrane proteins. This article is part of a Special Issue entitled: Membrane proteins edited by J.C. Gumbart and Sergei Noskov.

  20. Proteomic analysis of day-night variations in protein levels in the rat pineal gland

    DEFF Research Database (Denmark)

    Møller, Morten; Sparre, Thomas; Bache, Nicolai

    2007-01-01

    The pineal gland secretes the hormone melatonin. This secretion exhibits a circadian rhythm with a zenith during night and a nadir during day. We have performed proteome analysis of the superficial pineal gland in rats during daytime and nighttime. The proteins were extracted and subjected to 2-DE....... Of 1747 protein spots revealed by electrophoresis, densitometric analysis showed the up-regulation of 25 proteins during nighttime and of 35 proteins during daytime. Thirty-seven of the proteins were identified by MALDI-TOF MS. The proteins up-regulated during the night are involved in the Krebs cycle......, energy transduction, calcium binding, and intracellular transport. During the daytime, enzymes involved in glycolysis, electron transport, and also the Krebs cycle were up-regulated as well as proteins taking part in RNA binding and RNA processing. Our data show a prominent day-night variation...

  1. Gestational Protein Restriction Increases Cardiac Connexin 43 mRNA levels in male adult rat offspring.

    Science.gov (United States)

    Rossini, Kamila Fernanda; Oliveira, Camila Andrea de; Rebelato, Hércules Jonas; Esquisatto, Marcelo Augusto Marreto; Catisti, Rosana

    2017-07-01

    The dietary limitation during pregnancy influences the growth and development of the fetus and offspring and their health into adult life. The mechanisms underlying the adverse effects of gestational protein restriction (GPR) in the development of the offspring hearts are not well understood. The aim of this study was to evaluate the effects of GPR on cardiac structure in male rat offspring at day 60 after birth (d60). Pregnant Wistar rats were fed a normal-protein (NP, 17% casein) or low-protein (LP, 6% casein) diet. Blood pressure (BP) values from 60-day-old male offspring were measured by an indirect tail-cuff method using an electro sphygmomanometer. Hearts (d60) were collected for assessment of connexin 43 (Cx43) mRNA expression and morphological and morphometric analysis. LP offspring showed no difference in body weight, although they were born lighter than NP offspring. BP levels were significantly higher in the LP group. We observed a significant increase in the area occupied by collagen fibers, a decrease in the number of cardiomyocytes by 104 µm2, and an increase in cardiomyocyte area associated with an increased Cx43 expression. GPR changes myocardial levels of Cx43 mRNA in male young adult rats, suggesting that this mechanism aims to compensate the fibrotic process by the accumulation of collagen fibers in the heart interstitium. A limitação dietética durante a gravidez influencia o crescimento e desenvolvimento do feto e da prole e sua saúde na vida adulta. Os mecanismos subjacentes dos efeitos adversos da restrição proteica gestacional (RPG) no desenvolvimento dos corações da prole não são bem compreendidos. Avaliar os efeitos da RPG sobre a estrutura cardíaca em filhotes machos de ratas aos 60 dias após o nascimento (d60). Ratos fêmeas Wistar grávidas foram alimentadas com uma dieta de proteína normal (PN, 17% caseína) ou de baixa proteína (BP, caseína 6%). Os valores de pressão arterial (PA) de descendentes do sexo masculino de

  2. Effect of experimental design and protein substitution strategy on the outcome of feeding different levels of protein in primiparous dairy cows

    Science.gov (United States)

    Inferences about lactation responses to diet have been hypothesized to be affected by the use of change-over instead of continuous experimental designs. A direct test of this hypothesis has not been well studied. Additionally, when dietary protein level is changed it must occur through dilution with...

  3. In vivo quantitative proteomics of somatosensory cortical synapses shows which protein levels are modulated by sensory deprivation.

    Science.gov (United States)

    Butko, Margaret T; Savas, Jeffrey N; Friedman, Beth; Delahunty, Claire; Ebner, Ford; Yates, John R; Tsien, Roger Y

    2013-02-19

    Postnatal bilateral whisker trimming was used as a model system to test how synaptic proteomes are altered in barrel cortex by sensory deprivation during synaptogenesis. Using quantitative mass spectrometry, we quantified more than 7,000 synaptic proteins and identified 89 significantly reduced and 161 significantly elevated proteins in sensory-deprived synapses, 22 of which were validated by immunoblotting. More than 95% of quantified proteins, including abundant synaptic proteins such as PSD-95 and gephyrin, exhibited no significant difference under high- and low-activity rearing conditions, suggesting no tissue-wide changes in excitatory or inhibitory synaptic density. In contrast, several proteins that promote mature spine morphology and synaptic strength, such as excitatory glutamate receptors and known accessory factors, were reduced significantly in deprived synapses. Immunohistochemistry revealed that the reduction in SynGAP1, a postsynaptic scaffolding protein, was restricted largely to layer I of barrel cortex in sensory-deprived rats. In addition, protein-degradation machinery such as proteasome subunits, E2 ligases, and E3 ligases, accumulated significantly in deprived synapses, suggesting targeted synaptic protein degradation under sensory deprivation. Importantly, this screen identified synaptic proteins whose levels were affected by sensory deprivation but whose synaptic roles have not yet been characterized in mammalian neurons. These data demonstrate the feasibility of defining synaptic proteomes under different sensory rearing conditions and could be applied to elucidate further molecular mechanisms of sensory development.

  4. Evaluation of Lipid Profile and Proteins level among Infected Patients with Hepatitis B and C Viruses

    Directory of Open Access Journals (Sweden)

    Safa Mohamed Osman

    2015-10-01

    Full Text Available Examination of plasma lipid profile could be considered as a clinical index to reflect liver damage in the active phase of hepatitis. This study aims to assessment of serum of lipid profile and proteins in patients infected with HBV and HCV. In cross sectional study 80 subjects were enrolled in this study. 40 diagnosed with positive HBV or HCV and 40 healths apparently as control group, age ranging between 19-55 years. Lipid profile, TP and Alb were measured by using spectophotometric methods of full automated Mindray analyzer. The results showed HBV and HCV is common in youngest with 62.5% versus adults 37.5%. Also found significant increase in mean concentration of TG and LDL-C infected patients with (P-value 0.004 and 0.000 respectively. The results of TC, HDL-C, TP and Alb showed insignificant differences with P-value (0.346, 0.121, 0.328 respectively. Study concludes that, patients infected with HBV and HCV have higher level of TG and LDL-C, may indicate fatty liver as the results of hepatocellular damage.   

  5. Environmental load of wels (Silurus glanis fed by feeds of different protein levels

    Directory of Open Access Journals (Sweden)

    Gábor Beliczky

    2013-01-01

    Full Text Available The most dangerous effect of the environment on traditional and intensive fish farming is thereleased nutriment loaded effluent water. Intensity is increased and the nutriment intake by artificialfeeds cannot be utilized all by fish. Parts of these remain in the system in particulate or dissolved formand begin to accumulate, may cause insufficient water quality. During our experiment we examined theenvironmental effects of three different feeds of different protein levels. The declared crude proteincontents of feeds were 33%, 40% and 49%. At the beginning of our research, catfish were fed by feedsof 1% of bodyweight. We measured the inorganic dissolved nitrogen forms of water before addingnourishment and after in the 6th, 12th, 24th, 36th, 48th, 60th and 72nd hours. Before sampling wechecked the main physical and chemical parameters of water which are essential to analysis. With regardto the results, the concentrations of ammonium nitrogen were permanently increased in control and alsoin the other treatments during the 72h examined period.

  6. Meiotic regulation of TPX2 protein levels governs cell cycle progression in mouse oocytes.

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    Stéphane Brunet

    Full Text Available Formation of female gametes requires acentriolar spindle assembly during meiosis. Mitotic spindles organize from centrosomes and via local activation of the RanGTPase on chromosomes. Vertebrate oocytes present a RanGTP gradient centred on chromatin at all stages of meiotic maturation. However, this gradient is dispensable for assembly of the first meiotic spindle. To understand this meiosis I peculiarity, we studied TPX2, a Ran target, in mouse oocytes. Strikingly, TPX2 activity is controlled at the protein level through its accumulation from meiosis I to II. By RNAi depletion and live imaging, we show that TPX2 is required for spindle assembly via two distinct functions. It controls microtubule assembly and spindle pole integrity via the phosphorylation of TACC3, a regulator of MTOCs activity. We show that meiotic spindle formation in vivo depends on the regulation of at least a target of Ran, TPX2, rather than on the regulation of the RanGTP gradient itself.

  7. Leptin level in plasma of lactating buffaloes fed two diets with different energy and protein concentrations

    Directory of Open Access Journals (Sweden)

    A. Parmeggiani

    2011-03-01

    Full Text Available Leptin, a protein mainly secreted from the white adipocytes, has been shown to contribute to the regulation of energy metabolism, feeding behaviour and whole body energy balance. Moreover, leptin gene activity and leptin secretion are correlated with body adiposity and changes in food intake. Furthermore, leptin could also modulate endocrine response to changes in nutritional status and/or tissue sensitivity to hormones (Houseknecht et al., 1998; Romsos, 1998. Several factors are known to influence plasma leptin in rodents and humans: particularly it increases by body fatness, insulin, glucocorticoids, estrogens and decreases by food deprivation (Saladin et al., 1995; Ahima et al., 1996; Shimizu et al., 1997. These ones and several other observations have led to the hypothesis that leptin is a signal arising from adipose tissue, linked to the level of fat reserves and/or the nutritional status. This signal directly influences the central nervous system and peripheral organs, resulting in a better adaptation of body metabolism and physiological functions to the availability of metabolic energy...........

  8. Inhibition of fatty acid binding proteins elevates brain anandamide levels and produces analgesia.

    Directory of Open Access Journals (Sweden)

    Martin Kaczocha

    Full Text Available The endocannabinoid anandamide (AEA is an antinociceptive lipid that is inactivated through cellular uptake and subsequent catabolism by fatty acid amide hydrolase (FAAH. Fatty acid binding proteins (FABPs are intracellular carriers that deliver AEA and related N-acylethanolamines (NAEs to FAAH for hydrolysis. The mammalian brain expresses three FABP subtypes: FABP3, FABP5, and FABP7. Recent work from our group has revealed that pharmacological inhibition of FABPs reduces inflammatory pain in mice. The goal of the current work was to explore the effects of FABP inhibition upon nociception in diverse models of pain. We developed inhibitors with differential affinities for FABPs to elucidate the subtype(s that contributes to the antinociceptive effects of FABP inhibitors. Inhibition of FABPs reduced nociception associated with inflammatory, visceral, and neuropathic pain. The antinociceptive effects of FABP inhibitors mirrored their affinities for FABP5, while binding to FABP3 and FABP7 was not a predictor of in vivo efficacy. The antinociceptive effects of FABP inhibitors were mediated by cannabinoid receptor 1 (CB1 and peroxisome proliferator-activated receptor alpha (PPARα and FABP inhibition elevated brain levels of AEA, providing the first direct evidence that FABPs regulate brain endocannabinoid tone. These results highlight FABPs as novel targets for the development of analgesic and anti-inflammatory therapeutics.

  9. Signaling Pathways Related to Protein Synthesis and Amino Acid Concentration in Pig Skeletal Muscles Depend on the Dietary Protein Level, Genotype and Developmental Stages.

    Directory of Open Access Journals (Sweden)

    Yingying Liu

    Full Text Available Muscle growth is regulated by the homeostatic balance of the biosynthesis and degradation of muscle proteins. To elucidate the molecular interactions among diet, pig genotype, and physiological stage, we examined the effect of dietary protein concentration, pig genotype, and physiological stages on amino acid (AA pools, protein deposition, and related signaling pathways in different types of skeletal muscles. The study used 48 Landrace pigs and 48 pure-bred Bama mini-pigs assigned to each of 2 dietary treatments: lower/GB (Chinese conventional diet- or higher/NRC (National Research Council-protein diet. Diets were fed from 5 weeks of age to respective market weights of each genotype. Samples of biceps femoris muscle (BFM, type I and longissimus dorsi muscle (LDM, type II were collected at nursery, growing, and finishing phases according to the physiological stage of each genotype, to determine the AA concentrations, mRNA levels for growth-related genes in muscles, and protein abundances of mechanistic target of rapamycin (mTOR signaling pathway. Our data showed that the concentrations of most AAs in LDM and BFM of pigs increased (P<0.05 gradually with increasing age. Bama mini-pigs had generally higher (P<0.05 muscle concentrations of flavor-related AA, including Met, Phe, Tyr, Pro, and Ser, compared with Landrace pigs. The mRNA levels for myogenic determining factor, myogenin, myocyte-specific enhancer binding factor 2 A, and myostatin of Bama mini-pigs were higher (P<0.05 than those of Landrace pigs, while total and phosphorylated protein levels for protein kinase B, mTOR, and p70 ribosomal protein S6 kinases (p70S6K, and ratios of p-mTOR/mTOR, p-AKT/AKT, and p-p70S6K/p70S6K were lower (P<0.05. There was a significant pig genotype-dependent effect of dietary protein on the levels for mTOR and p70S6K. When compared with the higher protein-NRC diet, the lower protein-GB diet increased (P<0.05 the levels for mTOR and p70S6K in Bama mini-pigs, but

  10. Effect of dietary crude protein level and degradability on ruminal fermentation and nitrogen utilization in lactating dairy cows.

    Science.gov (United States)

    Hristov, A N; Etter, R P; Ropp, J K; Grandeen, K L

    2004-11-01

    The objectives of this experiment were to investigate the effects of two ruminally degradable protein (RDP) levels in diets containing similar ruminally undegradable protein (RUP) and metabolizable protein (MP) concentrations on ruminal fermentation, digestibility, and transfer of ruminal ammonia N into milk protein in dairy cows. Four ruminally and duodenally cannulated Holstein cows were allocated to two dietary treatments in a crossover design. The diets (adequate RDP [ARDP] and high RDP [HRDP]), had similar concentrations of RUP and MP, but differed in CP/RDP content. Ruminal ammonia was labeled with 15N and secretion of tracer in milk protein was determined for a period of 120 h. Ammonia concentration in the rumen tended to be greater (P = 0.06) with HRDP than with ARDP. Microbial N flow to the duodenum, ruminal digestibility of dietary nutrients, DMI, milk yield, fat content, and protein content and yield were not statistically different between diets. There was a tendency (P = 0.07) for increased urinary N excretion, and blood plasma and milk urea N concentrations were greater (P = 0.002 and P = 0.01, respectively) with HRDP compared with ARDP. Milk N efficiency was decreased (P = 0.01) by the HRDP diet. The cumulative secretion of ammonia 15N into milk protein, as a proportion of 15N dosed intraruminally, was greater (P = 0.003) with ARDP than with HRDP. The proportions of bacterial protein originating from ammonia N and milk protein originating from bacterial or ammonia N averaged 43, 61, and 26% and were not affected by diet. This experiment indicated that excess RDP in the diet of lactating dairy cows could not be efficiently utilized for microbial protein synthesis and was largely lost through urinary N excretion. At a similar MP supply, increased CP or RDP concentration of the diet would result in decreased efficiency of conversion of dietary N into milk protein and less efficient use of ruminal ammonia N for milk protein syntheses.

  11. In Situ Molecular Level Studies on Membrane Related Peptides and Proteins in Real Time Using Sum Frequency Generation Vibrational Spectroscopy

    Science.gov (United States)

    Ye, Shuji; Nguyen, Khoi Tan; Le Clair, Stéphanie V.; Chen, Zhan

    2009-01-01

    Sum frequency generation (SFG) vibrational spectroscopy has been demonstrated to be a powerful technique to study the molecular structures of surfaces and interfaces in different chemical environments. This review summarizes recent SFG studies on hybrid bilayer membranes and substrate-supported lipid monolayers and bilayers, the interaction between peptides/proteins and lipid monolayers/bilayers, and bilayer perturbation induced by peptides/proteins. To demonstrate the ability of SFG to determine the orientations of various secondary structures, studies on the interaction between different peptides/proteins (melittin, G proteins, almethicin, and tachyplesin I) and lipid bilayers are discussed. Molecular level details revealed by SFG in these studies show that SFG can provide a unique understanding on the interactions between a lipid monolayer/bilayer and peptides/proteins in real time, in situ and without any exogenous labeling. PMID:19306928

  12. Low-level lasers on microRNA and uncoupling protein 2 mRNA levels in human breast cancer cells

    Science.gov (United States)

    Canuto, K. S.; Teixeira, A. F.; Rodrigues, J. A.; Paoli, F.; Nogueira, E. M.; Mencalha, A. L.; Fonseca, A. S.

    2017-06-01

    MicroRNA is short non-coding RNA and is a mediator of post-transcriptional regulation of gene expression. In addition, uncoupling proteins (UCPs) regulate thermogenesis, metabolic and energy balance, and decrease reactive oxygen species production. Both microRNA and UCP2 expression can be altered in cancer cells. At low power, laser wavelength, frequency, fluence and emission mode deternube photobiological responses, which are the basis of low-level laser therapy. There are few studies on miRNA and UCP mRNA levels after low-level laser exposure on cancer cells. In this work, we evaluate the micrRNA (mir-106b and mir-15a) and UCP2 mRNA levels in human breast cancer cells exposed to low-level lasers. MDA-MB-231 human breast cancer cells were exposed to low-level red and infrared lasers, total RNA was extracted for cDNA synthesis and mRNA levels by real time quantitative polymerase chain reaction were evaluated. Data show that mir-106b and mir-15a relative levels are not altered, but UCP2 mRNA relative levels are increased in MDA-MB-231 human breast cancer cells exposed to low-level red and infrared lasers at fluences used in therapeutic protocols.

  13. 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.

  14. Discrepant NOXA (PMAIP1) transcript and NOXA protein levels: a potential Achilles' heel in mantle cell lymphoma.

    Science.gov (United States)

    Dengler, M A; Weilbacher, A; Gutekunst, M; Staiger, A M; Vöhringer, M C; Horn, H; Ott, G; Aulitzky, W E; van der Kuip, H

    2014-01-23

    Mantle cell lymphoma (MCL) is an aggressive lymphoid neoplasm with transient response to conventional chemotherapy. We here investigated the role of the Bcl-2 homology domain 3-only protein NOXA for life-death decision in MCL. Surprisingly, NOXA (PMAIP1) mRNA and NOXA protein levels were extremely discrepant in MCL cells: NOXA mRNA was found to be highly expressed whereas NOXA protein levels were low. Chronic active B-cell receptor signaling and to a minor degree cyclin D1 overexpression contributed to high NOXA mRNA expression levels in MCL cells. The phoshatidyl-inositol-3 kinase/AKT/mammalian target of rapamycin pathway was identified as the major downstream signaling pathway involved in the maintenance of NOXA gene expression. Interestingly, MCL cells adapt to this constitutive pro-apoptotic signal by extensive ubiquitination and rapid proteasomal degradation of NOXA protein (T½∼15-30 min). In addition to the proteasome inhibitor Bortezomib, we identified the neddylation inhibitor MLN4924 and the fatty acid synthase inhibitor Orlistat as potent inducers of NOXA protein expression leading to apoptosis in MCL. All inhibitors targeted NOXA protein turnover. In contrast to Bortezomib, MLN4924 and Orlistat interfered with the ubiquitination process of NOXA protein thereby offering new strategies to kill Bortezomib-resistant MCL cells. Our data, therefore, highlight a critical role of NOXA in the balance between life and death in MCL. The discrepancy between NOXA transcript and protein levels is essential for sensitivity of MCL to ubiquitin-proteasome system inhibitors and could therefore provide a druggable Achilles' heel of MCL cells.

  15. Level of heat shock proteins decreases in individuals carrying B-chromosomes in the grasshopper Eyprepocnemis plorans

    DEFF Research Database (Denmark)

    Teruel, M; Sørensen, Jesper Givskov; Loeschcke, Volker

    2010-01-01

    We analyzed the effect of B-chromosome presence on expression level of heat shock protein 70 (Hsp70) in cerebral ganglion and gonad in both males and females of the grasshopper Eyprepocnemis plorans. Two natural Spanish populations, Salobreña (Granada) and Torrox (Málaga) were assayed, the former......-chromosome effects in Torrox showed a dose-dependent pattern. The results point to an interesting interaction between B-chromosome and stress protein expression in reproductive tissue....

  16. Influence of periodontal therapy on C-reactive protein level: a systematic review and meta-analysis

    OpenAIRE

    Camila Oliveira Teixeira de Freitas; Isaac Suzart Gomes-Filho; Roberta Catapano Naves; Getúlio da Rocha Nogueira Filho; Simone Seixas da Cruz; Carlos Antonio de Souza Teles Santos; Leonardo Dunningham; Lituânia Fialho de Miranda; Mônica Dourado da Silva Barbosa

    2012-01-01

    The influence of oral infections, especially periodontal disease, on systemic diseases has been extensively discussed in the literature. Because periodontal disease is a persistent infection, it promotes an inflammatory response. C-reactive protein is a marker for inflammatory reactions that is frequently studied, since elevated levels of this protein are related to coronary events. OBJECTIVE: The aim of this study was to investigate the effect of periodontal therapy on reducing the serum lev...

  17. High level of serum cholesteryl ester transfer protein inactive hepatitis C virus infection

    Institute of Scientific and Technical Information of China (English)

    2016-01-01

    AIM To determine the significance of cholesteryl estertransfer protein (CETP) in lipoprotein abnormalities inchronic hepatitis C virus (HCV) infection.METHODS: We evaluated the significance of the serumconcentration of CETP in 110 Japanese patients withchronic HCV infection. Fifty-five patients had active HCVinfection, and HCV eradication had been achieved in55. The role of CETP in serum lipoprotein abnormalities,specifically, in triglyceride (TG) concentrations in thefour major classes of lipoproteins, was investigatedusing Pearson correlations in conjunction with multipleregression analysis and compared them between thosewith active HCV infection and those in whom eradicationhad been achieved.RESULTS: The serum CETP levels of patients withactive HCV infection were significantly higher than thoseof patients in whom HCV eradication was achieved(mean ± SD, 2.84 ± 0.69 μg/mL vs 2.40 ± 1.00 μg/mL, P = 0.008). In multiple regression analysis, HCVinfection status (active or eradicated) was an independentfactor significantly associated with the serumCETP level. TG concentrations in low-density lipoprotein(mean ± SD, 36.25 ± 15.28 μg/mL vs 28.14 ± 9.94μg/mL, P = 0.001) and high-density lipoprotein (HDL)(mean ± SD, 25.9 ± 7.34 μg/mL vs 17.17 ± 4.82μg/mL, P 〈 0.001) were significantly higher in patients with active HCV infection than in those in whom HCVeradication was achieved. The CETP level was stronglycorrelated with HDL-TG in patients with active HCVinfection (R = 0.557, P 〈 0.001), whereas CETP wasnot correlated with HDL-TG in patients in whom HCVeradication was achieved (R = -0.079, P = 0.56).CONCLUSION: Our results indicate that CETP playsa role in abnormalities of lipoprotein metabolism inpatients with chronic HCV infection.

  18. Effects of dietary crude protein level on odour from pig manure.

    Science.gov (United States)

    Le, P D; Aarnink, A J A; Jongbloed, A W; Peet-Schwering, C M C Van der; Ogink, N W M; Verstegen, M W A

    2007-06-01

    The objective of this study was to determine the effects of dietary crude protein (CP) level on odour emission, odour intensity, hedonic tone, and ammonia emission from pig manure and on manure composition (pH, total nitrogen, ammonium, volatile fatty acids, indolic, phenolic and sulphur-containing compounds). An experiment was conducted with growing pigs (n = 18) in a randomised complete-block design with three treatments in six blocks. Treatment groups were 12%, 15% and 18% CP diets. Barley was exchanged for soya-bean meal. Crystalline amino acids (AA) were included in the 12% CP diet up to the level of pigs' requirement; the same amount of AA was added to the 15% and 18% CP diets. Pigs with an initial body weight (BW) of 36.5 ± 3.4 kg (mean ± s.d.) were individually penned in partly slatted floor pens and offered a daily feed allowance of 2.8 × maintenance requirement for net energy (NE: 293 kJ/kg BW0.75). Feed was mixed with water, 1/2.5 (w/w). Faeces and urine of each pig were accumulated together in a separate manure pit under the slatted floor. After an adaptation period of 2 weeks, the manure pits were cleaned and manure was collected. In the 5th week of the collection period, air samples for odour and ammonia analyses, and manure samples were collected directly from each manure pit. Air samples were analysed for odour concentration and for hedonic value and intensity above odour detection threshold. Manure samples were analysed for volatile fatty acids, and indolic, phenolic and sulphurous compounds, ammonium and total nitrogen concentrations. Reducing dietary CP from 18% to 12% lowered odour emission ( P acids and cresols concentrations in the manure of pigs fed different dietary CP levels were similar. A reduction of dietary CP and at the same time providing essential AA is an option to reduce odour emission as well as ammonia emission from pig manure.

  19. Repeat use of human recombinant bone morphogenetic protein-2 for second level lumbar arthrodesis.

    Science.gov (United States)

    Singh, Kern; Dumonski, Mark; Stanley, Tom; Ponnappan, Ravi; Phillips, Frank M

    2011-02-01

    Prospective randomized controlled animal model. The purpose of this study is to determine whether the readministration of human recombinant bone morphogenetic protein-2 (rhBMP-2) induces an immune response and inhibits successful fusion in repeat posterolateral spinal surgery. Little research has been performed on the effectiveness or immunoreactivity of rhBMP-2 (Infuse, Medtronic, Memphis, TN) in the context of its reuse in posterolateral fusion spinal surgery at adjacent levels. A total of 34 New Zealand White rabbits underwent posterior intertransverse process fusion with the use of rhBMP-2 delivered on an absorbable collagen sponge (rhBMP-2/ACS). Two rabbits were killed early leaving 32 total rabbits. Serologic studies (Type I bovine collagen and rhBMP-2 antibodies) were obtained at 2-week intervals throughout the experiment. At 10 weeks, posteroanterior radiographs confirmed solid fusion masses in all rabbits. The 32 rabbits were randomly separated into 2 groups of 16, and each group underwent an adjacent level, bilateral intertransverse process fusion with either rhBMP-2/ACS or iliac crest. There was no statistical difference in fusion rates with repeat use of rhBMP-2 (n = 15/16, 94%) or iliac crest (n = 11/16, 69%) (P = 0.17) at the adjacent level. Four rabbits (n = 4/32, 13%) developed rhBMP-2 antibodies. Of these 4 rabbits, 1 developed anti-rhBMP antibodies after the first exposure and 3 developed antibodies after the second surgery. Eight rabbits (n = 8/32, 25%) developed collagen antibodies with 7 rabbits developing antibodies after the first exposure and 1 rabbit developing antibodies after the second exposure. The development of antibodies did not effect fusion rates. No rabbit demonstrated evidence of a systemic or anaphylactic reaction to repeat exposure to rhBMP-2. rhBMP-2 appears to be successful in promoting intertransverse fusions when used in both primary and repeat fusion environments. The infrequent development of antibodies to rhBMP-2 after

  20. Influence of periodontal therapy on C-reactive protein level: a systematic review and meta-analysis

    Directory of Open Access Journals (Sweden)

    Camila Oliveira Teixeira de Freitas

    2012-02-01

    Full Text Available The influence of oral infections, especially periodontal disease, on systemic diseases has been extensively discussed in the literature. Because periodontal disease is a persistent infection, it promotes an inflammatory response. C-reactive protein is a marker for inflammatory reactions that is frequently studied, since elevated levels of this protein are related to coronary events. OBJECTIVE: The aim of this study was to investigate the effect of periodontal therapy on reducing the serum levels of C-reactive protein, by means of a systematic review of the literature and meta-analysis. MATERIAL AND METHODS: A systematic review of the English-language literature was conducted in the PUBMED-MEDLINE database, using the key words "periodontal disease", "C-reactive protein", "periodontal therapy" and "periodontal treatment", in accordance with the terms for Medical Subject Headings (MeSH, to evaluate the effect of periodontal therapy on C-reactive protein levels. A qualitative analysis of studies of randomized clinical trial design was performed using CONSORT, with subsequent meta-analysis. RESULTS: The literature search initially retrieved 46 potentially relevant studies available in the databases. From these, in accordance with the inclusion criteria, only 11 were selected, of which only 4 fulfilled the criteria of randomized clinical trial design. According to CONSORT, the studies evaluated generally presented good quality with regard to the criteria analyzed. Through meta-analysis, the reduction in mean levels of C-reactive protein (-0.231; p=0.000 after introducing periodontal treatment was estimated. The result was statistically significant, without evidence of heterogeneity between the studies (p=0.311. CONCLUSIONS: The findings indicated that non-surgical periodontal treatment had a positive effect with regard to reduction of the serum levels of C-reactive protein.

  1. Influence of periodontal therapy on C-reactive protein level: a systematic review and meta-analysis

    Science.gov (United States)

    de FREITAS, Camila Oliveira Teixeira; GOMES-FILHO, Isaac Suzart; NAVES, Roberta Catapano; NOGUEIRA FILHO, Getúlio da Rocha; da CRUZ, Simone Seixas; SANTOS, Carlos Antonio de Souza Teles; DUNNINGHAM, Leonardo; de MIRANDA, Lituânia Fialho; BARBOSA, Mônica Dourado da Silva

    2012-01-01

    The influence of oral infections, especially periodontal disease, on systemic diseases has been extensively discussed in the literature. Because periodontal disease is a persistent infection, it promotes an inflammatory response. C-reactive protein is a marker for inflammatory reactions that is frequently studied, since elevated levels of this protein are related to coronary events. Objective The aim of this study was to investigate the effect of periodontal therapy on reducing the serum levels of C-reactive protein, by means of a systematic review of the literature and meta-analysis. Material and Methods A systematic review of the English-language literature was conducted in the PUBMED-MEDLINE database, using the key words "periodontal disease", "C-reactive protein", "periodontal therapy" and "periodontal treatment", in accordance with the terms for Medical Subject Headings (MeSH), to evaluate the effect of periodontal therapy on C-reactive protein levels. A qualitative analysis of studies of randomized clinical trial design was performed using CONSORT, with subsequent meta-analysis. Results The literature search initially retrieved 46 potentially relevant studies available in the databases. From these, in accordance with the inclusion criteria, only 11 were selected, of which only 4 fulfilled the criteria of randomized clinical trial design. According to CONSORT, the studies evaluated generally presented good quality with regard to the criteria analyzed. Through meta-analysis, the reduction in mean levels of C-reactive protein (-0.231; p=0.000) after introducing periodontal treatment was estimated. The result was statistically significant, without evidence of heterogeneity between the studies (p=0.311). Conclusions The findings indicated that non-surgical periodontal treatment had a positive effect with regard to reduction of the serum levels of C-reactive protein. PMID:22437670

  2. Plasma protein C levels in immunocompromised septic patients are significantly lower than immunocompetent septic patients: a prospective cohort study

    Directory of Open Access Journals (Sweden)

    Bird Robert

    2009-10-01

    Full Text Available Abstract Introduction Activated Protein C [APC] improves outcome in immunocompetent patients with severe sepsis particularly in those who are perceived to have high mortality risk. Before embarking on a trial of APC administration in immunocompromised septic patients, a preliminary study on plasma levels of protein C in this cohort is essential. Objective To assess serum Protein C concentrations in immunocompromised patients as compared to immunocompetent patients during sepsis, severe sepsis, septic shock and recovery. Methods Prospective cohort study in a tertiary hospital. Patients satisfying inclusion criteria were enrolled after informed consent. Clinical variables were noted with sample collection when patients met criteria for sepsis, severe sepsis, septic shock and recovery. Protein C levels were measured using monoclonal antibody based fluorescence immunoassay. Results Thirty one patients participated in this study (22 immunocompromised, 9 immunocompetent. Protein C levels were found to be significantly lower in the immunocompromised group compared to the immunocompetent group, particularly observed in severe sepsis [2.27 (95% CI: 1.63-2.9 vs 4.19 (95% CI: 2.87-5.52 mcg/ml] (p = 0.01 and sepsis [2.59 (95% CI: 1.98-3.21 vs 3.64 (95% CI: 2.83-4.45 mcg/ml] (p = 0.03. SOFA scores were similar in both the groups across sepsis, severe sepsis and septic shock categories. Protein C levels improved significantly in recovery (p = 0.001 irrespective of immune status. Conclusion Protein C levels were significantly lower in immunocompromised patients when compared to immunocompetent patients in severe sepsis and sepsis categories. Our study suggests a plausible role for APC in severely septic immunocompromised patients which need further elucidation.

  3. Ski protein levels increase during in vitro progression of HPV16-immortalized human keratinocytes and in cervical cancer

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Yi; Pirisi, Lucia [Department of Pathology, Microbiology and Immunology, School of Medicine, University of South Carolina, Columbia, SC (United States); Creek, Kim E., E-mail: creekk@sccp.sc.edu [Department of Drug Discovery and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina, Columbia, SC (United States)

    2013-09-15

    We compared the levels of the Ski oncoprotein, an inhibitor of transforming growth factor-beta (TGF-β) signaling, in normal human keratinocytes (HKc), HPV16 immortalized HKc (HKc/HPV16), and differentiation resistant HKc/HPV16 (HKc/DR) in the absence and presence of TGF-β. Steady-state Ski protein levels increased in HKc/HPV16 and even further in HKc/DR, compared to HKc. TGF-β treatment of HKc, HKc/HPV16, and HKc/DR dramatically decreased Ski. TGF-β-induced Ski degradation was delayed in HKc/DR. Ski and phospho-Ski protein levels are cell cycle dependent with maximal Ski expression and localization to centrosomes and mitotic spindles during G2/M. ShRNA knock down of Ski in HKc/DR inhibited cell proliferation. More intense nuclear and cytoplasmic Ski staining and altered Ski localization were found in cervical cancer samples compared to adjacent normal tissue in a cervical cancer tissue array. Overall, these studies demonstrate altered Ski protein levels, degradation and localization in HPV16-transformed human keratinocytes and in cervical cancer. - Highlights: • Ski oncoprotein levels increase during progression of HPV16-transformed cells. • Ski and phospho-Ski protein levels are cell cycle dependent. • Ski knock-down in HPV16-transformed keratinocytes inhibited cell proliferation. • Cervical cancer samples overexpress Ski.

  4. Ezrin Binds to DEAD-Box RNA Helicase DDX3 and Regulates Its Function and Protein Level.

    Science.gov (United States)

    Çelik, Haydar; Sajwan, Kamal P; Selvanathan, Saravana P; Marsh, Benjamin J; Pai, Amrita V; Kont, Yasemin Saygideger; Han, Jenny; Minas, Tsion Z; Rahim, Said; Erkizan, Hayriye Verda; Toretsky, Jeffrey A; Üren, Aykut

    2015-09-01

    Ezrin is a key regulator of cancer metastasis that links the extracellular matrix to the actin cytoskeleton and regulates cell morphology and motility. We discovered a small-molecule inhibitor, NSC305787, that directly binds to ezrin and inhibits its function. In this study, we used a nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS-MS)-based proteomic approach to identify ezrin-interacting proteins that are competed away by NSC305787. A large number of the proteins that interact with ezrin were implicated in protein translation and stress granule dynamics. We validated direct interaction between ezrin and the RNA helicase DDX3, and NSC305787 blocked this interaction. Downregulation or long-term pharmacological inhibition of ezrin led to reduced DDX3 protein levels without changes in DDX3 mRNA. Ectopic overexpression of ezrin in low-ezrin-expressing osteosarcoma cells caused a notable increase in DDX3 protein levels. Ezrin inhibited the RNA helicase activity of DDX3 but increased its ATPase activity. Our data suggest that ezrin controls the translation of mRNAs preferentially with a structured 5' untranslated region, at least in part, by sustaining the protein level of DDX3 and/or regulating its function. Therefore, our findings suggest a novel function for ezrin in regulation of gene translation that is distinct from its canonical role as a cytoskeletal scaffold at the cell membrane.

  5. Improvement of proteolytic efficiency towards low-level proteins by an antifouling surface of alumina gel in a microchannel.

    Science.gov (United States)

    Liu, Yun; Wang, Huixiang; Liu, Qingping; Qu, Haiyun; Liu, Baohong; Yang, Pengyuan

    2010-11-07

    A microfluidic reactor has been developed for rapid enhancement of protein digestion by constructing an alumina network within a poly(ethylene terephthalate) (PET) microchannel. Trypsin is stably immobilized in a sol-gel network on the PET channel surface after pretreatment, which produces a protein-resistant interface to reduce memory effects, as characterized by X-ray fluorescence spectrometry and electroosmotic flow. The gel-derived network within a microchannel provides a large surface-to-volume ratio stationary phase for highly efficient proteolysis of proteins existing both at a low level and in complex extracts. The maximum reaction rate of the encapsulated trypsin reactor, measured by kinetic analysis, is much faster than in bulk solution. Due to the microscopic confinement effect, high levels of enzyme entrapment and the biocompatible microenvironment provided by the alumina gel network, the low-level proteins can be efficiently digested using such a microreactor within a very short residence time of a few seconds. The on-chip microreactor is further applied to the identification of a mixture of proteins extracted from normal mouse liver cytoplasm sample via integration with 2D-LC-ESI-MS/MS to show its potential application for large-scale protein identification.

  6. A Low Protein Diet Alters Bone Material Level Properties and the Response to In Vitro Repeated Mechanical Loading

    Directory of Open Access Journals (Sweden)

    Victor Dubois-Ferrière

    2014-01-01

    Full Text Available Low protein intake is associated with an alteration of bone microstructure and material level properties. However, it remains unknown whether these alterations of bone tissue could influence the response to repeated mechanical loading. The authors investigated the in vitro effect of repeated loading on bone strength in humeri collected from 20 6-month-old female rats pair-fed with a control (15% casein or an isocaloric low protein (2.5% casein diet for 10 weeks. Bone specimens were cyclically loaded in three-point bending under load control for 2000 cycles. Humeri were then monotonically loaded to failure. The load-displacement curve of the in vitro cyclically loaded humerus was compared to the contralateral noncyclically loaded humerus and the influence of both protein diets. Material level properties were also evaluated through a nanoindentation test. Cyclic loading decreased postyield load and plastic deflection in rats fed a low protein diet, but not in those on a regular diet. Bone material level properties were altered in rats fed a low protein diet. This suggests that bone biomechanical alterations consequent to cyclic loading are more likely to occur in rats fed a low protein diet than in control animals subjected to the same in vitro cyclic loading regimen.

  7. Proteomic Investigation of Protein Profile Changes and Amino Acid Residue Level Modification in Cooked Lamb Meat: The Effect of Boiling.

    Science.gov (United States)

    Yu, Tzer-Yang; Morton, James D; Clerens, Stefan; Dyer, Jolon M

    2015-10-21

    Hydrothermal treatment (heating in water) is a common method of general food processing and preparation. For red-meat-based foods, boiling is common; however, how the molecular level effects of this treatment correlate to the overall food properties is not yet well-understood. The effects of differing boiling times on lamb meat and the resultant cooking water were here examined through proteomic evaluation. The longer boiling time was found to result in increased protein aggregation involving particularly proteins such as glyceraldehyde-3-phosphate dehydrogenase, as well as truncation in proteins such as in α-actinin-2. Heat-induced protein backbone cleavage was observed adjacent to aspartic acid and asparagine residues. Side-chain modifications of amino acid residues resulting from the heating, including oxidation of phenylalanine and formation of carboxyethyllysine, were characterized in the cooked samples. Actin and myoglobin bands from the cooked meat per se remained visible on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, even after significant cooking time. These proteins were also found to be the major source of observed heat-induced modifications. This study provides new insights into molecular-level modifications occurring in lamb meat proteins during boiling and a protein chemistry basis for better understanding the effect of this common treatment on the nutritional and functional properties of red-meat-based foods.

  8. 2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum.

    Directory of Open Access Journals (Sweden)

    Andrea Kreidenweiss

    Full Text Available Analysis of gene function in Plasmodium falciparum, the most important human malaria parasite, is restricted by the lack of robust and simple reverse genetic tools. Approaches to manipulate protein levels post-translationally are powerful tools to study protein-off effects especially in the haploid malaria parasite where genetic knockouts of essential genes are lethal. We investigated if the auxin-inducible degron system is functional in P. falciparum and found that degron-tagged yellow fluorescent protein levels were efficiently reduced upon addition of auxin which otherwise had no effect on parasite viability. The genetic components required in this conditional approach were co-expressed in P. falciparum by applying the small peptide 2A. 2A is a self-processing peptide from Foot-And-Mouth Disease virus that allows the whole conditional system to be accommodated on a single plasmid vector and ensures stoichiometric expression levels.

  9. Genome-wide association study of CSF levels of 59 alzheimer's disease candidate proteins: significant associations with proteins involved in amyloid processing and inflammation.

    Directory of Open Access Journals (Sweden)

    John S K Kauwe

    2014-10-01

    Full Text Available Cerebrospinal fluid (CSF 42 amino acid species of amyloid beta (Aβ42 and tau levels are strongly correlated with the presence of Alzheimer's disease (AD neuropathology including amyloid plaques and neurodegeneration and have been successfully used as endophenotypes for genetic studies of AD. Additional CSF analytes may also serve as useful endophenotypes that capture other aspects of AD pathophysiology. Here we have conducted a genome-wide association study of CSF levels of 59 AD-related analytes. All analytes were measured using the Rules Based Medicine Human DiscoveryMAP Panel, which includes analytes relevant to several disease-related processes. Data from two independently collected and measured datasets, the Knight Alzheimer's Disease Research Center (ADRC and Alzheimer's Disease Neuroimaging Initiative (ADNI, were analyzed separately, and combined results were obtained using meta-analysis. We identified genetic associations with CSF levels of 5 proteins (Angiotensin-converting enzyme (ACE, Chemokine (C-C motif ligand 2 (CCL2, Chemokine (C-C motif ligand 4 (CCL4, Interleukin 6 receptor (IL6R and Matrix metalloproteinase-3 (MMP3 with study-wide significant p-values (p<1.46×10-10 and significant, consistent evidence for association in both the Knight ADRC and the ADNI samples. These proteins are involved in amyloid processing and pro-inflammatory signaling. SNPs associated with ACE, IL6R and MMP3 protein levels are located within the coding regions of the corresponding structural gene. The SNPs associated with CSF levels of CCL4 and CCL2 are located in known chemokine binding proteins. The genetic associations reported here are novel and suggest mechanisms for genetic control of CSF and plasma levels of these disease-related proteins. Significant SNPs in ACE and MMP3 also showed association with AD risk. Our findings suggest that these proteins/pathways may be valuable therapeutic targets for AD. Robust associations in cognitively normal

  10. Post-transcriptional regulation of ITGB6 protein levels in damaged skeletal muscle

    OpenAIRE

    Ducceschi, Melissa; Clifton, Lisa G.; Stimpson, Stephen A.; Billin, Andrew N.

    2014-01-01

    We have identified integrin beta 6 (Itgb6) as a transcript highly enriched in skeletal muscle. This finding is unexpected because Itgb6 is typically associated with epithelial expression domains in normal tissue. Further we find that ITGB6 protein expression in muscle is post-transcriptionally regulated. Uninjured muscle expresses Itgb6 RNA but no ITGB6 protein is detectable. Muscle injury induces ITGB6 protein accumulation rapidly post-injury in myofibers adjacent to the site of injury. As r...

  11. High-level synthesis of a heterologous milk protein in the mammary glands of transgenic swine.

    OpenAIRE

    Wall, R J; Pursel, V G; Shamay, A; McKnight, R A; Pittius, C W; Hennighausen, L

    1991-01-01

    The whey acidic protein (WAP) is a major milk protein in mice, rats, and rabbits but has not been found in milk of livestock including swine. To determine whether mammary gland regulatory elements from the WAP gene function across species boundaries and whether it is possible to qualitatively alter milk protein composition, we introduced the mouse WAP gene into the genome of swine. Three lines of transgenic swine were analyzed, and mouse WAP was detected in milk from all lactating females at ...

  12. Plasma surfactant protein D levels and the relation to body mass index in a chinese population

    DEFF Research Database (Denmark)

    Zhao, X M; Wu, Y P; Wei, R

    2007-01-01

    Surfactant protein D (SP-D) is a member of the collectin family and is an important component of the pulmonary innate host defence. The protein has a widespread distribution in the human body and is present in multiple epithelia, in endothelium and in blood. Various studies have looked at the rel......Surfactant protein D (SP-D) is a member of the collectin family and is an important component of the pulmonary innate host defence. The protein has a widespread distribution in the human body and is present in multiple epithelia, in endothelium and in blood. Various studies have looked...

  13. Glutamine metabolism in uricotelic species: variation in skeletal muscle glutamine synthetase, glutaminase, glutamine levels and rates of protein synthesis.

    Science.gov (United States)

    Watford, Malcolm; Wu, Guoyao

    2005-04-01

    High intracellular glutamine levels have been implicated in promoting net protein synthesis and accretion in mammalian skeletal muscle. Little is known regarding glutamine metabolism in uricotelic species but chicken breast muscle exhibits high rates of protein accretion and would be predicted to maintain high glutamine levels. However, chicken breast muscle expresses high glutaminase activity and here we report that chicken breast muscle also expresses low glutamine synthetase activity (0.07+/-0.01 U/g) when compared to leg muscle (0.50+/-0.04 U/g). Free glutamine levels were 1.38+/-0.09 and 9.69+/-0.12 nmol/mg wet weight in breast and leg muscles of fed chickens, respectively. Glutamine levels were also lower in dove breast muscle (4.82+/-0.35 nmol/mg wet weight) when compared to leg muscle (16.2+/-1.0 nmol/mg wet weight) and much lower (1.80+/-0.46 nmol/mg wet weight) in lizard leg muscle. In fed chickens, rates of fractional protein synthesis were higher in leg than in breast muscle, and starvation (48 h) resulted in a decrease in both glutamine content and rate of protein synthesis in leg muscle. Thus, although tissue-specific glutamine metabolism in uricotelic species differs markedly from that in ureotelic animals, differences in rates of skeletal muscle protein synthesis are associated with corresponding differences in intramuscular glutamine content.

  14. Gamma glutamyltransferase levels and its association with high sensitive C-reactive protein in patients with acute coronary syndromes

    Science.gov (United States)

    Emiroglu, Mehmet Yunus; Esen, Özlem Batukan; Bulut, Mustafa; Karapinar, Hekim; Kaya, Zekeriya; Akcakoyun, Mustafa; Kargin, Ramazan; Aung, Soe Moe; Alızade, Elnur; Pala, Selcuk; Esen, Ali Metin

    2010-01-01

    Background: Elevated Gamma-glutamyltransferase (GGT) level is independently correlated with conditions associatedwith increased atherosclerosis, such as obesity, elevated serum cholesterol, high blood pressure and myocardial infarction. It is also demonstrated that serum gamma-glutamyltransferase activity is an independent risk factor for myocardial infarction and cardiac death in patients with coronary artery disease. Although the relationship between gamma-glutamyltransferase and coronary artery disease has been reported, not many studies have shown the relationship between changes ofgamma-glutamyltransferase in acute coronary syndromes and a well established coronary risk factor high sensitive C-reactive protein. (hs-CRP). Aims: In this study, how gamma-glutamyltransferase levels changed in acute coronary syndromes and its relationship with high sensitive C-reactive protein if any were studied. Patients & Methods: This trial was carried out at Kosuyolu Cardiovascular Training and Research Hospital and Van Yuksek Ihtisas Hospital, Turkey. 219 patients (177 males and 42 females) presenting with acute coronary syndrome, and 51 control subjects between September 2007 and September 2008 were included in the study. Serum gamma-glutamyltransferase, high sensitive C-reactive protein, serum lipoprotein levels and troponin I were determined. Results: Serum gamma-glutamyltransferase and high sensitive C-reactive protein levels were higher in acute coronary syndrome patients compared to control. There was also correlation between gamma-glutamyltransferase and high sensitive C-reactive protein levels. Conclusion: Serum gamma-glutamyltransferase and high sensitive C-reactive protein levels were higher in acute coronary syndrome patients. In subgroup analyses, the higher difference with Non-ST elevation myocardial infarction and ST elevation myocardial infarction groups than unstable angina oectoris group proposes a relationship between gamma-glutamyltransferase and severity

  15. Gamma glutamyltransferase levels and its association with high sensitive C-reactive protein in patients with acute coronary syndromes

    Directory of Open Access Journals (Sweden)

    Mehmet Yunus Emiroglu

    2010-07-01

    Full Text Available Background: Elevated Gamma-glutamyltransferase (GGT level is independently correlated with conditions associatedwith increased atherosclerosis, such as obesity, elevated serum cholesterol, high blood pressure and myocardial infarction. It is also demonstrated that serum gamma-glutamyltransferase activity is an independent risk factor for myocardial infarction and cardiac death in patients with coronary artery disease. Although the relationship between gamma-glutamyltransferase and coronary artery disease has been reported, not many studies have shown the relationship between changes ofgamma-glutamyltransferase in acute coronary syndromes and a well established coronary risk factor high sensitive C-reactive protein. (hs-CRP. Aims: In this study, how gamma-glutamyltransferase levels changed in acute coronary syndromes and its relationship with high sensitive C-reactive protein if any were studied. Patients & Methods:This trial was carried out at Kosuyolu Cardiovascular Training and Research Hospital and Van Yuksek Ihtisas Hospital, Turkey. 219 patients (177 males and 42 females presenting with acute coronary syndrome, and 51 control subjects between September 2007 and September 2008 were included in the study. Serum gamma-glutamyltransferase, high sensitive C-reactive protein, serum lipoprotein levels and troponin I were determined. Results: Serum gamma-glutamyltransferase and high sensitive C-reactive protein levels were higher in acute coronary syndrome patients compared to control. There was also correlation between gamma-glutamyltransferase and high sensitive C-reactive protein levels. Conclusion: Serum gamma-glutamyltransferase and high sensitive C-reactive protein levels were higher in acute coronary syndrome patients. In subgroup analyses, the higher difference with Non-ST elevation myocardial infarction and ST elevation myocardial infarction groups than unstable angina oectoris group proposes a relationship between gamma

  16. Gamma glutamyltransferase levels and its association with high sensitive C-reactive protein in patients with acute coronary syndromes

    Directory of Open Access Journals (Sweden)

    Mehmet Yunus Emiroglu

    2010-01-01

    Full Text Available Background: Elevated Gamma-glutamyltransferase (GGT level is independently correlated with conditions associatedwith increased atherosclerosis, such as obesity, elevated serum cholesterol, high blood pressure and myocardial infarction. It is also demonstrated that serum gamma-glutamyltransferase activity is an independent risk factor for myocardial infarction and cardiac death in patients with coronary artery disease. Although the relationship between gamma-glutamyltransferase and coronary artery disease has been reported, not many studies have shown the relationship between changes ofgamma-glutamyltransferase in acute coronary syndromes and a well established coronary risk factor high sensitive C-reactive protein. (hs-CRP. Aims: In this study, how gamma-glutamyltransferase levels changed in acute coronary syndromes and its relationship with high sensitive C-reactive protein if any were studied. Patients & Methods: This trial was carried out at Kosuyolu Cardiovascular Training and Research Hospital and Van Yuksek Ihtisas Hospital, Turkey. 219 patients (177 males and 42 females presenting with acute coronary syndrome, and 51 control subjects between September 2007 and September 2008 were included in the study. Serum gamma-glutamyltransferase, high sensitive C-reactive protein, serum lipoprotein levels and troponin I were determined. Results: Serum gamma-glutamyltransferase and high sensitive C-reactive protein levels were higher in acute coronary syndrome patients compared to control. There was also correlation between gamma-glutamyltransferase and high sensitive C-reactive protein levels. Conclusion: Serum gamma-glutamyltransferase and high sensitive C-reactive protein levels were higher in acute coronary syndrome patients. In subgroup analyses, the higher difference with Non-ST elevation myocardial infarction and ST elevation myocardial infarction groups than unstable angina oectoris group proposes a relationship between gamma

  17. Icariin upregulates phosphorylated cyclic adenosine monophosphate response element binding protein levels in the hippocampus of the senescence- accelerated mouse

    Institute of Scientific and Technical Information of China (English)

    Zhanwei Zhang; Ting Zhang; Keli Dong

    2012-01-01

    At 8 weeks after intragastric administration of icariin to senescence-accelerated mice (P8 strain), Morris water maze results showed that escape latency was shortened, and the number of platform crossings was increased. Immunohistochemical staining and western blot assay detected signifi-cantly increased levels of cyclic adenosine monophosphate response element binding protein. These results suggest that icariin upregulates phosphorylated cyclic adenosine monophosphate response element binding protein levels and improves learning and memory functions in hippo-campus of the senescence-accelerated mouse.

  18. Protein levels of genes encoded on chromosome 21 in fetal Down Syndrome brain (Part V): overexpression of phosphatidyl-inositol-glycan class P protein (DSCR5).

    Science.gov (United States)

    Ferrando-Miguel, R; Cheon, M S; Lubec, G

    2004-06-01

    Down Syndrome (DS, trisomy 21) is the most common genetic cause of mental retardation. The completed sequencing of genes encoded on chromosome 21 provides excellent basic information, however the molecular mechanisms leading to the phenotype of DS remain to be elucidated. Although overexpression of chromosome 21 encoded genes has been documented information at the protein expression level is mandatory as it is the proteins that carry out function. We therefore decided to evaluated expression level of seven proteins whose genes are encoded on chromosome 21: DSCR4, DSCR5, DSCR6; KIR4.2, GIRK2, KCNE1 and KCNE2 in fetal cortex brain of DS and controls at the early second trimester of pregnancy by Western blotting. beta-actin and neuron specific enolase (NSE) were used to normalise cell loss and neuronal loss. DSCR5 (PIG-P), a component of glycosylphosphatidylinositol- N-acetylglucosaminyltransferase (GPI-GnT), was overexpressed about twofold, even when levels were normalised with NSE. DSCR6 was overexpressed in addition but when normalised versus NSE, levels were comparable to controls. DSCR4 was not detectable in fetal brain. Potassium channels KIR4.2 and GIRK2 were comparable between DS and controls, whereas KCNE1 and KCNE2 were not detectable. Quantification of these proteins encoded on chromosome 21 revealed that not all gene products of the DS critical region are overexpressed in DS brain early in life, indicating that the DS phenotype cannot be simply explained by the gene dosage effect hypothesis. Overexpression of PIG-P (DSCR5) may lead to or represent impaired glycosylphosphatidylinositol- N-acetylglucosaminyltransferase mediated posttranslational modifications and subsequent anchoring of proteins to the plasma membrane.

  19. Serum levels of advanced glycation endproducts and other markers of protein damage in early diabetic nephropathy in type 1 diabetes.

    Directory of Open Access Journals (Sweden)

    Bruce A Perkins

    Full Text Available OBJECTIVE: To determine the role of markers of plasma protein damage by glycation, oxidation and nitration in microalbuminuria onset or subsequent decline of glomerular filtration rate (termed "early GFR decline" in patients with type 1 diabetes. METHODS: From the 1(st Joslin Kidney Study, we selected 30 patients with longstanding normoalbuminuria and 55 patients with new onset microalbuminuria. Patients with microalbuminuria had 8-12 years follow-up during which 33 had stable GFR and 22 early GFR decline. Mean baseline GFR(CYSTATIN C was similar between the three groups. Glycation, oxidation and nitration markers were measured in protein and ultrafiltrate at baseline by liquid chromatography-tandem mass spectrometry using the most reliable methods currently available. RESULTS: Though none were significantly different between patients with microalbuminuria with stable or early GFR decline, levels of 6 protein damage adduct residues of plasma protein and 4 related free adducts of plasma ultrafiltrate were significantly different in patients with microalbuminuria compared to normoalbuminuria controls. Three protein damage adduct residues were decreased and 3 increased in microalbuminuria while 3 free adducts were decreased and one increased in microalbuminuria. The most profound differences were of N-formylkynurenine (NFK protein adduct residue and N(ω-carboxymethylarginine (CMA free adduct in which levels were markedly lower in microalbuminuria (P<0.001 for both. CONCLUSIONS: Complex processes influence levels of plasma protein damage and related proteolysis product free adducts in type 1 diabetes and microalbuminuria. The effects observed point to the possibility that patients who have efficient mechanisms of disposal of damaged proteins might be at an increased risk of developing microalbuminuria but not early renal function decline. The findings support the concept that the mechanisms responsible for microalbuminuria may differ from the

  20. Fusion protein-based biofilm fabrication composed of recombinant azurin–myoglobin for dual-level biomemory application

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Taek [Research Institute for Basic Science, Sogang University, Heukseok-dong, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Chung, Yong-Ho; Yoon, Jinho [Department of Chemical and Biomolecular Engineering, Sogang University, Heukseok-dong, Dongjak-gu, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742 (Korea, Republic of); Min, Junhong [School of Integrative Engineering, Chung-Ang University, Heukseok-dong, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Choi, Jeong-Woo, E-mail: jwchoi@sogang.ac.kr [Department of Chemical and Biomolecular Engineering, Sogang University, Heukseok-dong, Dongjak-gu, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742 (Korea, Republic of)

    2014-11-30

    Graphical abstract: - Highlights: • We developed the fusion protein-based biofilm on the inorganic surface. • For making the fusion protein, the recombinant azurin and the myoglobin was conjugated by the native chemical ligation method. • The developed fusion protein shows unique electrochemical property. • The proposed fusion protein biofilm appears to be a good method for dual-level biomemory device. - Abstract: In the present study, a fusion protein-based biofilm composed of a recombinant azurin–myoglobin (Azu-Myo) has been developed and confirmed its original electrochemical property for dual-level biomemory device application. For this purpose, the azurin was modified with cysteine residues for direct immobilization and conjugation. Then, the recombinant azurin was conjugated with the myoglobin via a sulfo-SMCC bifunctional linker using the chemical ligation method (CLM). The SDS-PAGE and UV–vis spectroscopy were performed to examine the fusion protein conjugates. The prepared Azu-Myo fusion protein was self-assembled onto Au substrate for the biofilm fabrication. Then, the atomic force microscopy (AFM) was used to confirm the immobilization and the surface-enhanced Raman spectroscopy (SERS) was carried out to the surface analysis. Also, the cyclic voltammetry (CV) was carried out to observe an electrochemical property of fabricated biofilm. As a result, the two pair of redox potential values was obtained for dual-level biomemory device application. Then, the dual-level biomemory function was verified by the multi-potential chronoamperometry (MPCA). The results indicate a new fabrication method and material combination for advances in bioelectronic device development.

  1. Overexpression of hydroxynitrile lyase in cassava roots elevates protein and free amino acids while reducing residual cyanogen levels.

    Directory of Open Access Journals (Sweden)

    Narayanan N Narayanan

    Full Text Available Cassava is the major source of calories for more than 250 million Sub-Saharan Africans, however, it has the lowest protein-to-energy ratio of any major staple food crop in the world. A cassava-based diet provides less than 30% of the minimum daily requirement for protein. Moreover, both leaves and roots contain potentially toxic levels of cyanogenic glucosides. The major cyanogen in cassava is linamarin which is stored in the vacuole. Upon tissue disruption linamarin is deglycosylated by the apolplastic enzyme, linamarase, producing acetone cyanohydrin. Acetone cyanohydrin can spontaneously decompose at pHs >5.0 or temperatures >35°C, or is enzymatically broken down by hydroxynitrile lyase (HNL to produce acetone and free cyanide which is then volatilized. Unlike leaves, cassava roots have little HNL activity. The lack of HNL activity in roots is associated with the accumulation of potentially toxic levels of acetone cyanohydrin in poorly processed roots. We hypothesized that the over-expression of HNL in cassava roots under the control of a root-specific, patatin promoter would not only accelerate cyanogenesis during food processing, resulting in a safer food product, but lead to increased root protein levels since HNL is sequestered in the cell wall. Transgenic lines expressing a patatin-driven HNL gene construct exhibited a 2-20 fold increase in relative HNL mRNA levels in roots when compared with wild type resulting in a threefold increase in total root protein in 7 month old plants. After food processing, HNL overexpressing lines had substantially reduced acetone cyanohydrin and cyanide levels in roots relative to wild-type roots. Furthermore, steady state linamarin levels in intact tissues were reduced by 80% in transgenic cassava roots. These results suggest that enhanced linamarin metabolism contributed to the elevated root protein levels.

  2. Overexpression of hydroxynitrile lyase in cassava roots elevates protein and free amino acids while reducing residual cyanogen levels.

    Science.gov (United States)

    Narayanan, Narayanan N; Ihemere, Uzoma; Ellery, Claire; Sayre, Richard T

    2011-01-01

    Cassava is the major source of calories for more than 250 million Sub-Saharan Africans, however, it has the lowest protein-to-energy ratio of any major staple food crop in the world. A cassava-based diet provides less than 30% of the minimum daily requirement for protein. Moreover, both leaves and roots contain potentially toxic levels of cyanogenic glucosides. The major cyanogen in cassava is linamarin which is stored in the vacuole. Upon tissue disruption linamarin is deglycosylated by the apolplastic enzyme, linamarase, producing acetone cyanohydrin. Acetone cyanohydrin can spontaneously decompose at pHs >5.0 or temperatures >35°C, or is enzymatically broken down by hydroxynitrile lyase (HNL) to produce acetone and free cyanide which is then volatilized. Unlike leaves, cassava roots have little HNL activity. The lack of HNL activity in roots is associated with the accumulation of potentially toxic levels of acetone cyanohydrin in poorly processed roots. We hypothesized that the over-expression of HNL in cassava roots under the control of a root-specific, patatin promoter would not only accelerate cyanogenesis during food processing, resulting in a safer food product, but lead to increased root protein levels since HNL is sequestered in the cell wall. Transgenic lines expressing a patatin-driven HNL gene construct exhibited a 2-20 fold increase in relative HNL mRNA levels in roots when compared with wild type resulting in a threefold increase in total root protein in 7 month old plants. After food processing, HNL overexpressing lines had substantially reduced acetone cyanohydrin and cyanide levels in roots relative to wild-type roots. Furthermore, steady state linamarin levels in intact tissues were reduced by 80% in transgenic cassava roots. These results suggest that enhanced linamarin metabolism contributed to the elevated root protein levels.

  3. Metabolic protein interactions in Bacillus subtilis studied at the single cell level

    NARCIS (Netherlands)

    Detert Oude Weme, Ruud Gerardus Johannes

    2015-01-01

    We have investigated protein-protein interactions in live Bacillus subtilis cells (a bacterium). B. subtilis’ natural habitat is the soil and the roots of plants, but also the human microbiota. B. subtilis is used worldwide as a model organism. Unlike eukaryotic cells, bacteria do not have organelle

  4. Revealing Atomic-Level Mechanisms of Protein Allostery with Molecular Dynamics Simulations.

    Directory of Open Access Journals (Sweden)

    Samuel Hertig

    2016-06-01

    Full Text Available Molecular dynamics (MD simulations have become a powerful and popular method for the study of protein allostery, the widespread phenomenon in which a stimulus at one site on a protein influences the properties of another site on the protein. By capturing the motions of a protein's constituent atoms, simulations can enable the discovery of allosteric binding sites and the determination of the mechanistic basis for allostery. These results can provide a foundation for applications including rational drug design and protein engineering. Here, we provide an introduction to the investigation of protein allostery using molecular dynamics simulation. We emphasize the importance of designing simulations that include appropriate perturbations to the molecular system, such as the addition or removal of ligands or the application of mechanical force. We also demonstrate how the bidirectional nature of allostery-the fact that the two sites involved influence one another in a symmetrical manner-can facilitate such investigations. Through a series of case studies, we illustrate how these concepts have been used to reveal the structural basis for allostery in several proteins and protein complexes of biological and pharmaceutical interest.

  5. Revealing Atomic-Level Mechanisms of Protein Allostery with Molecular Dynamics Simulations.

    Science.gov (United States)

    Hertig, Samuel; Latorraca, Naomi R; Dror, Ron O

    2016-06-01

    Molecular dynamics (MD) simulations have become a powerful and popular method for the study of protein allostery, the widespread phenomenon in which a stimulus at one site on a protein influences the properties of another site on the protein. By capturing the motions of a protein's constituent atoms, simulations can enable the discovery of allosteric binding sites and the determination of the mechanistic basis for allostery. These results can provide a foundation for applications including rational drug design and protein engineering. Here, we provide an introduction to the investigation of protein allostery using molecular dynamics simulation. We emphasize the importance of designing simulations that include appropriate perturbations to the molecular system, such as the addition or removal of ligands or the application of mechanical force. We also demonstrate how the bidirectional nature of allostery-the fact that the two sites involved influence one another in a symmetrical manner-can facilitate such investigations. Through a series of case studies, we illustrate how these concepts have been used to reveal the structural basis for allostery in several proteins and protein complexes of biological and pharmaceutical interest.

  6. The Impact of a Pure Protein Load on the Glucose Levels in Type 1 Diabetes Patients Treated with Insulin Pumps

    Directory of Open Access Journals (Sweden)

    Tomasz Klupa

    2015-01-01

    Full Text Available We aimed to estimate the impact of ingestion of a pure protein load on the glucose levels in T1DM patients treated with insulin pumps. We examined 10 T1DM patients (6 females, mean age—32.3 years, mean HbA1c—6.8% treated with insulin pumps equipped with a continuous glucose monitoring system (CGMS. In Phase I, baseline insulin infusion was optimized to minimize the differences in fasting glucose levels to less than 30 mg/dL between any two time points between 9 a.m. and 3 p.m. In Phase II, the patients were exposed to single pure protein load. CGMS record was performed and the glucose pattern was defined for 6 hours of each phase. The maximal glucose level increment was similar for the entire duration of the fasting and the protein load test (26.6 versus 27.6 mg/dL, resp., P<0.78. There was only a borderline difference in change between baseline versus 6th hour glucose (12.5 and 19.0 mg/dL, P=0.04. Glucose variability, assessed by standard deviation of mean glucose levels, was 36.4 and 37.9 mg/dL, respectively (P=0.01. The administration of a pure protein load does not seem to have a clinically significant impact on glucose levels in T1DM patients treated with insulin pumps.

  7. The impact of a pure protein load on the glucose levels in type 1 diabetes patients treated with insulin pumps.

    Science.gov (United States)

    Klupa, Tomasz; Benbenek-Klupa, Teresa; Matejko, Bartlomiej; Mrozinska, Sandra; Malecki, Maciej T

    2015-01-01

    We aimed to estimate the impact of ingestion of a pure protein load on the glucose levels in T1DM patients treated with insulin pumps. We examined 10 T1DM patients (6 females, mean age-32.3 years, mean HbA1c-6.8%) treated with insulin pumps equipped with a continuous glucose monitoring system (CGMS). In Phase I, baseline insulin infusion was optimized to minimize the differences in fasting glucose levels to less than 30 mg/dL between any two time points between 9 a.m. and 3 p.m. In Phase II, the patients were exposed to single pure protein load. CGMS record was performed and the glucose pattern was defined for 6 hours of each phase. The maximal glucose level increment was similar for the entire duration of the fasting and the protein load test (26.6 versus 27.6 mg/dL, resp., P < 0.78). There was only a borderline difference in change between baseline versus 6th hour glucose (12.5 and 19.0 mg/dL, P = 0.04). Glucose variability, assessed by standard deviation of mean glucose levels, was 36.4 and 37.9 mg/dL, respectively (P = 0.01). The administration of a pure protein load does not seem to have a clinically significant impact on glucose levels in T1DM patients treated with insulin pumps.

  8. The Impact of a Pure Protein Load on the Glucose Levels in Type 1 Diabetes Patients Treated with Insulin Pumps

    Science.gov (United States)

    Klupa, Tomasz; Benbenek-Klupa, Teresa; Matejko, Bartlomiej; Mrozinska, Sandra; Malecki, Maciej T.

    2015-01-01

    We aimed to estimate the impact of ingestion of a pure protein load on the glucose levels in T1DM patients treated with insulin pumps. We examined 10 T1DM patients (6 females, mean age—32.3 years, mean HbA1c—6.8%) treated with insulin pumps equipped with a continuous glucose monitoring system (CGMS). In Phase I, baseline insulin infusion was optimized to minimize the differences in fasting glucose levels to less than 30 mg/dL between any two time points between 9 a.m. and 3 p.m. In Phase II, the patients were exposed to single pure protein load. CGMS record was performed and the glucose pattern was defined for 6 hours of each phase. The maximal glucose level increment was similar for the entire duration of the fasting and the protein load test (26.6 versus 27.6 mg/dL, resp., P < 0.78). There was only a borderline difference in change between baseline versus 6th hour glucose (12.5 and 19.0 mg/dL, P = 0.04). Glucose variability, assessed by standard deviation of mean glucose levels, was 36.4 and 37.9 mg/dL, respectively (P = 0.01). The administration of a pure protein load does not seem to have a clinically significant impact on glucose levels in T1DM patients treated with insulin pumps. PMID:25767510

  9. Enhanced levels of glutathione and protein glutathiolation in rat tongue epithelium during 4-NQO-induced carcinogenesis.

    Science.gov (United States)

    Huang, Zhishan; Komninou, Despina; Kleinman, Wayne; Pinto, John T; Gilhooly, Elaine M; Calcagnotto, Ana; Richie, John P

    2007-04-01

    High glutathione (GSH) levels are commonly found in oral tumors and are thought to play an important role in tumorigenesis. While posttranslational binding of GSH to cellular proteins (protein glutathiolation) has recently been recognized as an important redox-sensitive regulatory mechanism, no data currently exist on this process during carcinogenesis. Our goal was to determine the effects of 4-nitroquinoline-N-oxide (4-NQO)-induced carcinogenesis on tongue levels of protein-bound and free GSH and related thiols in the rat. Male F-344 rats (6 weeks of age) were administered either 4-NQO (20 ppm) in drinking water or tap water alone (controls) for 8 weeks. Twenty-four weeks after cessation of 4-NQO, squamous cell carcinomas of the tongue were observed in all rats. The levels of both free and bound GSH in tumors, as well as in adjacent tissues, were 2- to 3-fold greater than in tongue epithelium from control rats (p tongue tissues from control rats (ptongue tissues from 4-NQO treated vs. control rats (p<0.05). Altogether, these results suggest that protein glutathiolation, together with GSH and GSSG levels, are induced during oral carcinogenesis in the rat possibly as a result of enhanced levels of oxidative stress.

  10. Different levels of protein and energy on physiological behavior and performance of European quail in the Brazilian semiarid

    Directory of Open Access Journals (Sweden)

    Thaiz Lamy Alves Ribeiro

    2016-06-01

    Full Text Available This study was designed to evaluate the effect of different levels of metabolizable energy and crude protein on the respiratory rate and the rectal and surface temperatures of 360 European quail (Coturnix coturnix coturnix raised on the Brazilian semiarid. The animals were distributed in a completely randomized design with a 3 per 2 factorial arrangement consisting of three protein levels (20, 21 and 22% and two energy levels (3050 and 3150 kcal/kg. We measured the physiological parameters of the quail at 42 days of age. The average rectal temperature noted in this experiment was 41.8°C, which was within the normal range. A significant effect (P <0.05 was verified at the levels of 21 and 22% of crude protein and 3150 kcal/kg. We found that the animals presented the highest respiratory rate in this setting, which is evidence that the panting mechanism results in maintaining thermal homeostasis. The surface temperature of the animals was lower for the 3050 kcal/kg diets. Levels of 21 and 22% of crude protein and 3150 kcal/kg of metabolizable energy were associated with the largest weight gain and the best feed conversion.

  11. Crude protein levels in diets containing pelleted concentrate for lactating goats: intake, digestibility, milk production and composition

    Directory of Open Access Journals (Sweden)

    Edileusa de Jesus dos Santos

    2015-08-01

    Full Text Available This study aimed to evaluate the effects of crude protein (100, 130, 160 and 190 g kg-1 of dry matter of diets composed of 200 g kg-1 of Tifton 85 grass hay and 800 g kg-1 of pelleted concentrate on intake, nutrient digestibility, production and composition of milk in lactating goats. Eight female Saanen goats with 42.7 ± 1.43 kg and 57.7 ± 7.37 days of lactation and milk production of 2 ± 0.22 kg at the beginning of the experiment were housed in individual 1.32 × 3.10 m stalls and distributed into two 4 × 4-balanced Latin squares. Intake of dry matter, organic matter, crude protein, neutral detergent fiber corrected for ash and protein, ether extract and total digestible nutrients showed a quadratic effect, with maximum intake of 2.030; 2.000; 305; 769; 55 and 1.574 g day-1 at the levels of 140.7; 140.8; 189.2; 140.9; 144.9 e 142.7 g kg-1 DM, respectively. Digestibility of dry matter, organic matter, crude protein, non-fibrous carbohydrates, ether extract and total digestible nutrient level varied linearly, with increases estimated at 0.54; 0.50, 2.02, 0.49, 0.80 and 0.63 g/100g for each percentage unit of protein added to the diet, respectively. Milk production was affected, with increase of 0.54 g for each 1% crude protein added to the diet. Milk lactose level decreased linearly, unlike the fat level, which increased linearly. Protein level showed a quadratic behavior, with a maximum of 36.7 g per kg of milk at the level of 160.5 g per kg of DM. It is recommended to use crude protein between 135 g and 150 g per kg of dry matter of diets consisting of 800 g of pelleted concentrate (composed of soybean meal replacing the alfalfa hay as protein source per kg of DM for lactating goats producing 2 kg of milk per day.

  12. Association of interleukin-6 and C-reactive protein genetic polymorphisms levels with venous thromboembolism

    Institute of Scientific and Technical Information of China (English)

    Ailiman Mahemuti; Kailibinuer Abudureheman; Xiaimuxikamaier Aihemaiti; HU Xue-mei; XIA Yu-ning; TANG Bao-peng; Halmurat Upur

    2012-01-01

    Background Increased levels of interleukin-6(IL-6)and C-reactive protein(CRP)have been reported in patients with venous thromboembolisms(VTE).However,prospective studies did not confirm an association between IL-6,CRP and their polymorphism with the risk of V-TE.Methods One hundred and forty patients(including 66 males and 74 females,mean age(55.55±17.11)years)and one hundred and sixty controls(including 74 males and 86 females,mean age(56.58±12.24)years)were involved.An enzyme linked immunosorbent assay(ELISA)method was used for detecting the serum levels of inflammatory factors IL-6 and CRP in both groups.Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)was used for analyzing the distribution of polymorphisms at the-572C/G and-597G/A sites of the promoter of the IL-6 gene and at 1059G/C of the CRP gene.Results Serum levels of IL-6 and CRP were significantly higher in the VTE group than in the control group(P<0.05).The frequencies of-572C/G promoter polymorphisms CC,CG,and GG in the IL-6 gene were found to be 34%,48%,and 18%,respectively,and the derived allele frequencies for the C and G alleles were 58% and 42%.There was a significant difference in the-572C/G promoter polymorphisms between the VTE group and control group(P<0.05).For the-597G/A polymorphism,individuals all carried the GG and GA type;AA genotypes were not detected.The frequency of the GG,GC,and CC genotypes at the CRP1059G/C promoter was 87.57%,7.86% and 3.57% in VTE group,while 86.25%,10%,and 3.75% in control group,respectively.The frequency of G and C alleles at CRP 1059G/C was 91.43%and 8.57% in VTE group and 91.56% and 8.44% in the control group.The results showed that there was no statistically significant difference of 1059G/C genotype and mutation frequency of the allele between the VTE group and control group(P>0.05).Multiple Logistic regression analysis showed CC homozygotes of the IL-6-572G/C,body mass index(BMI),and CRP

  13. C-Reactive Protein, Sialic Acid and Adenosine Deaminase Levels in Serum and Pleural Fluid from Patients with Pleural Effusion

    Science.gov (United States)

    Kim, Ji Woon; Yang, In Ae; Oh, Eun A; Rhyoo, Young Gun; Jang, Young Ho; Ryang, Dong Wook; Yoo, JooYong

    1988-01-01

    Laboratory analysis of pleural fluids is essential to determine underlying diseases. The authors evaluated the clinical significance of C-reactive protein (C-RP), sialic acid (SA), and adenosine deaminase (ADA) determinations in sera and pleural fluids from 37 patients with pleural effusion. (FP12)C-RP and sialic acid levels and ADA activities were higher in exudates than in transudates of pleural fluids. Serum and pleural fluid C-RP levels were high in patients with pyothorax. Determinations of serum sialic acid and the pleural fluid to serum ratio were useful for the differential diagnosis of pulmonary tuberculosis and malignancy. ADA activities of pleural fluid and serum are useful for the differentiation of malignancy from tuberculosis and nonspecific pyothorax. C-RP concentrations of pleural fluid correlated to serum levels. However, concentrations of sialic acid and ADA activities were not correlated to serum levels and only correlated to protein concentrations of pleural fluids. PMID:3154188

  14. meta-Tyrosine induces modification of reactive nitrogen species level, protein nitration and nitrosoglutathione reductase in tomato roots.

    Science.gov (United States)

    Krasuska, Urszula; Andrzejczak, Olga; Staszek, Paweł; Borucki, Wojciech; Gniazdowska, Agnieszka

    2017-08-01

    A non-protein amino acid (NPAA) - meta-Tyrosine (m-Tyr), is a harmful compound produced by fescue roots. Young (3-4 days old) tomato (Solanum lycopersicum L.) seedlings were supplemented for 24-72 h with m-Tyr (50 or 250 μM) inhibiting root growth by 50 or 100%, without lethal effect. Fluorescence of DAF-FM and APF derivatives was determined to show reactive nitrogen species (RNS) localization and level in roots of tomato plants. m-Tyr-induced restriction of root elongation growth was related to formation of nitrated proteins described as content of 3-nitrotyrosine. Supplementation with m-Tyr enhanced superoxide radicals generation in extracts of tomato roots and stimulated protein nitration. It correlated well to increase of fluorescence of DAF-FM derivatives, and transiently stimulated fluorescence of APF derivatives corresponding respectively to NO and ONOO(-) formation. Alterations in RNS formation induced by m-Tyr were linked to metabolism of nitrosoglutathione (GSNO). Activity of nitrosoglutatione reductase (GSNOR), catalyzing degradation of GSNO was enhanced by long term plant supplementation with m-Tyr, similarly as protein abundance, while transcripts level were only slightly altered by tested NPAA. We conclude, that although in animal cells m-Tyr is considered as a marker of oxidative stress, its secondary mode of action in tomato plants involves perturbation in RNS formation, alteration in GSNO metabolism and modification of protein nitration level. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. The effect of varying protein levels on blood chemistry, food consumption, and behavior of captive seaducks

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    Wells-Berlin, A. M.; Perry, M.C.; Olsen, G.H.

    2005-01-01

    The Chesapeake Bay is a primary wintering area for scoters and the long-tailed ducks (Clangia hyemalis) that migrate along the Atlantic Flyway. Recently, the Chesapeake Bay had undergone an ecosystem shift and little is known about how this is affecting the seaduck populations. We are determining what are the preferred food sources of the seaducks wintering on the Bay and analyzing the factors influencing prey selection whether it is prey composition, energy assimilated, prey availability, or a combination of any or all of these factors. We have established a captive colony of surf (Melanitta perspicillata) and white-winged scoters (Melanitta fusca) as well as long-tailed ducks at Patuxent Wildlife Research Center to allow us to examine these factors in a more controlled environment. This project contains a multitude of experiments and the resultant data will be compiled into a compartmental model on the feeding ecology of seaducks wintering on the Bay. The first experiment entailed feeding groups of each species (four ducks per pen of equal sex ratio, if possible, and four pens per species) three diets varying in percent protein levels from November to February. Each diet was randomly assigned to each pen and the amount of food consumed was recorded each day. New feed was given when all existing food was consumed. Behavioral trials and blood profiles were completed on all study birds to determine the effects of the varying diets. There were no significant differences in food consumption, blood chemistry, and behavior detected at the 5% level among the diets for all three species of interest. There was a seasonal effect determined based on the food consumption data for white-winged scoters, but not for surf scoters or long-tailed ducks. The blood profiles of the surf scoters were compared to blood profiles of wild surf scoters and a there was no difference detected at the 5% level. As a health check of the ducks an aspergillosis test was run on the blood obtained

  16. Plasma Monocyte Chemoattractant Protein-1 Level as a Predictor of the Severity of Community-Acquired Pneumonia.

    Science.gov (United States)

    Yong, Kok-Khun; Chang, Jer-Hwa; Chien, Ming-Hsien; Tsao, Shih-Ming; Yu, Ming-Chih; Bai, Kuan-Jen; Tsao, Thomas Chang-Yao; Yang, Shun-Fa

    2016-01-29

    Monocyte chemoattractant protein (MCP)-1 increases in the serum of immunocompetent patients with community-acquired pneumonia (CAP). However, the correlation between the circulating level of MCP-1 and severity of CAP remains unclear. This study investigated differential changes in the plasma MCP-1 levels of patients with CAP before and after an antibiotic treatment and further analyzes the association between the CAP severity and MCP-1 levels. We measured the plasma MCP-1 levels of 137 patients with CAP and 74 healthy controls by using a commercial enzyme-linked immunosorbent assay. Upon initial hospitalization, Acute Physiology and Chronic Health Evaluation II (APACHE II); confusion, urea level, respiratory rate, blood pressure, and age of >64 years (CURB-65); and pneumonia severity index (PSI) scores were determined for assessing the CAP severity in these patients. The antibiotic treatment reduced the number of white blood cells (WBCs) and neutrophils as well as the level of C-reactive protein (CRP) and MCP-1. The plasma MCP-1 level, but not the CRP level or WBC count, correlated with the CAP severity according to the PSI (r = 0.509, p < 0.001), CURB-65 (r = 0.468, p < 0.001), and APACHE II (r = 0.360, p < 0.001) scores. We concluded that MCP-1 levels act in the development of CAP and are involved in the severity of CAP.

  17. Blood profile of proteins and steroid hormones predicts weight change after weight loss with interactions of dietary protein level and glycemic index.

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    Ping Wang

    Full Text Available BACKGROUND: Weight regain after weight loss is common. In the Diogenes dietary intervention study, high protein and low glycemic index (GI diet improved weight maintenance. OBJECTIVE: To identify blood predictors for weight change after weight loss following the dietary intervention within the Diogenes study. DESIGN: Blood samples were collected at baseline and after 8-week low caloric diet-induced weight loss from 48 women who continued to lose weight and 48 women who regained weight during subsequent 6-month dietary intervention period with 4 diets varying in protein and GI levels. Thirty-one proteins and 3 steroid hormones were measured. RESULTS: Angiotensin I converting enzyme (ACE was the most important predictor. Its greater reduction during the 8-week weight loss was related to continued weight loss during the subsequent 6 months, identified by both Logistic Regression and Random Forests analyses. The prediction power of ACE was influenced by immunoproteins, particularly fibrinogen. Leptin, luteinizing hormone and some immunoproteins showed interactions with dietary protein level, while interleukin 8 showed interaction with GI level on the prediction of weight maintenance. A predictor panel of 15 variables enabled an optimal classification by Random Forests with an error rate of 24±1%. A logistic regression model with independent variables from 9 blood analytes had a prediction accuracy of 92%. CONCLUSIONS: A selected panel of blood proteins/steroids can predict the weight change after weight loss. ACE may play an important role in weight maintenance. The interactions of blood factors with dietary components are important for personalized dietary advice after weight loss. REGISTRATION: ClinicalTrials.gov NCT00390637.

  18. High expression level of bone degrading proteins as a possible inducer of osteolytic features in pigmented villonodular synovitis.

    Science.gov (United States)

    Geldyyev, Aman; Koleganova, Nadezda; Piecha, Grzegorz; Sueltmann, Holger; Finis, Katharina; Ruschaupt, Markus; Poustka, Annemarie; Gross, Marie-Luise; Berger, Irina

    2007-10-08

    Protein expression of osteopontin (OPN), osteoprotegerin (OPG), bone sialoprotein (BSP), osteocalcin (OC), RANKL and PTHrP was determined by use of immunohistochemical analysis on tissue arrays (48 cases of PVNS, 20 cases of active (a-RA), non-active rheumatoid arthritis (na-RA), and osteoarthritis (OA)). Additionally, gene expression was analysed using complimentary DNA (cDNA) microarrays. All PVNS cases showed a higher level of both protein and gene expression of RANKL, OPN and BSP in comparison with OA cases. Expression of OPG was not significantly different in PVNS compared to OA. The RANKL/OPG expression ratio was significantly higher in PVNS than in OA. High expressions level of proteins involved in bone degradation in PVNS may promote an intra-osseous propagation of the lesion. This evidence suggests that PVNS might respond to treatment using specific inhibitors of RANKL, OPN and BSP.

  19. Human oligodendroglial cells express low levels of C1 inhibitor and membrane cofactor protein mRNAs

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    McGeer Patrick L

    2004-08-01

    Full Text Available Abstract Background Oligodendrocytes, neurons, astrocytes, microglia, and endothelial cells are capable of synthesizing complement inhibitor proteins. Oligodendrocytes are vulnerable to complement attack, which is particularly observed in multiple sclerosis. This vulnerability may be related to a deficiency in their ability to express complement regulatory proteins. Methods This study compared the expression level of complement inhibitor mRNAs by human oligodendrocytes, astrocytes and microglia using semi-quantitative RT-PCR. Results Semi-quantitative RT-PCR analysis showed that C1 inhibitor (C1-inh mRNA expression was dramatically lower in oligodendroglial cells compared with astrocytes and microglia. The mRNA expression level of membrane cofactor protein (MCP by oligodendrocytes was also significantly lower than for other cell types. Conclusion The lower mRNA expression of C1-inh and MCP by oligodendrocytes could contribute to their vulnerability in several neurodegenerative and inflammatory diseases of the central nervous system.

  20. Comparison of serum leptin, glucose, total cholesterol and total protein levels in fertile and repeat breeder cows

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    Saime Guzel

    2014-12-01

    Full Text Available In the present study we measured serum glucose, leptin, total cholesterol and total protein concentrations in repeat breeder cows and compared them with fertile cows. For this aim, 20 repeat breeder cows and 20 fertile cows were used as material. Repeat breeder cows were found to have lower levels of leptin and glucose as compared with fertile ones. No significant differences in total cholesterol and total protein levels were observed between the two groups. No significant correlation of leptin with glucose, total cholesterol and total protein was observed in fertile and repeat breeder cows. Low concentrations of glucose and leptin can have some effects on reproductive problems as repeat breeder and help to understand potential mechanisms impairing fertility in repeat breeder cows.