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Sample records for residual tak activity

  1. HTLV-1 Tax-mediated TAK1 activation involves TAB2 adapter protein

    International Nuclear Information System (INIS)

    Yu Qingsheng; Minoda, Yasumasa; Yoshida, Ryoko; Yoshida, Hideyuki; Iha, Hidekatsu; Kobayashi, Takashi; Yoshimura, Akihiko; Takaesu, Giichi

    2008-01-01

    Human T cell leukemia virus type 1 (HTLV-1) Tax is an oncoprotein that plays a crucial role in the proliferation and transformation of HTLV-1-infected T lymphocytes. It has recently been reported that Tax activates a MAPKKK family, TAK1. However, the molecular mechanism of Tax-mediated TAK1 activation is not well understood. In this report, we investigated the role of TAK1-binding protein 2 (TAB2) in Tax-mediated TAK1 activation. We found that TAB2 physically interacts with Tax and augments Tax-induced NF-κB activity. Tax and TAB2 cooperatively activate TAK1 when they are coexpressed. Furthermore, TAK1 activation by Tax requires TAB2 binding as well as ubiquitination of Tax. We also found that the overexpression of TRAF2, 5, or 6 strongly induces Tax ubiquitination. These results suggest that TAB2 may be critically involved in Tax-mediated activation of TAK1 and that NF-κB-activating TRAF family proteins are potential cellular E3 ubiquitin ligases toward Tax

  2. TAK1 is activated in the myocardium after pressure overload and is sufficient to provoke heart failure in transgenic mice

    Science.gov (United States)

    Zhang, D.; Gaussin, V.; Taffet, G. E.; Belaguli, N. S.; Yamada, M.; Schwartz, R. J.; Michael, L. H.; Overbeek, P. A.; Schneider, M. D.

    2000-01-01

    The transforming-growth-factor-beta-activated kinase TAK1 is a member of the mitogen-activated protein kinase kinase kinase family, which couples extracellular stimuli to gene transcription. The in vivo function of TAK1 is not understood. Here, we investigated the potential involvement of TAK1 in cardiac hypertrophy. In adult mouse myocardium, TAK1 kinase activity was upregulated 7 days after aortic banding, a mechanical load that induces hypertrophy and expression of transforming growth factor beta. An activating mutation of TAK1 expressed in myocardium of transgenic mice was sufficient to produce p38 mitogen-activated protein kinase phosphorylation in vivo, cardiac hypertrophy, interstitial fibrosis, severe myocardial dysfunction, 'fetal' gene induction, apoptosis and early lethality. Thus, TAK1 activity is induced as a delayed response to mechanical stress, and can suffice to elicit myocardial hypertrophy and fulminant heart failure.

  3. MEK and TAK1 Regulate Apoptosis in Colon Cancer Cells with KRAS-Dependent Activation of Proinflammatory Signaling.

    Science.gov (United States)

    McNew, Kelsey L; Whipple, William J; Mehta, Anita K; Grant, Trevor J; Ray, Leah; Kenny, Connor; Singh, Anurag

    2016-12-01

    MEK inhibitors have limited efficacy in treating RAS-RAF-MEK pathway-dependent cancers due to feedback pathway compensation and dose-limiting toxicities. Combining MEK inhibitors with other targeted agents may enhance efficacy. Here, codependencies of MEK, TAK1, and KRAS in colon cancer were investigated. Combined inhibition of MEK and TAK1 potentiates apoptosis in KRAS-dependent cells. Pharmacologic studies and cell-cycle analyses on a large panel of colon cancer cell lines demonstrate that MEK/TAK1 inhibition induces cell death, as assessed by sub-G 1 accumulation, in a distinct subset of cell lines. Furthermore, TAK1 inhibition causes G 2 -M cell-cycle blockade and polyploidy in many of the cell lines. MEK plus TAK1 inhibition causes reduced G 2 -M/polyploid cell numbers and additive cytotoxic effects in KRAS/TAK1-dependent cell lines as well as a subset of BRAF-mutant cells. Mechanistically, sensitivity to MEK/TAK1 inhibition can be conferred by KRAS and BMP receptor activation, which promote expression of NF-κB-dependent proinflammatory cytokines, driving tumor cell survival and proliferation. MEK/TAK1 inhibition causes reduced mTOR, Wnt, and NF-κB signaling in TAK1/MEK-dependent cell lines concomitant with apoptosis. A Wnt/NF-κB transcriptional signature was derived that stratifies primary tumors into three major subtypes: Wnt-high/NF-κB-low, Wnt-low/NF-κB-high and Wnt-high/NF-κB-high, designated W, N, and WN, respectively. These subtypes have distinct characteristics, including enrichment for BRAF mutations with serrated carcinoma histology in the N subtype. Both N and WN subtypes bear molecular hallmarks of MEK and TAK1 dependency seen in cell lines. Therefore, N and WN subtype signatures could be utilized to identify tumors that are most sensitive to anti-MEK/TAK1 therapeutics. This study describes a potential therapeutic strategy for a subset of colon cancers that are dependent on oncogenic KRAS signaling pathways, which are currently difficult to

  4. Procarcinogenic effects of cyclosporine A are mediated through the activation of TAK1/TAB1 signaling pathway

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Jianmin; Walsh, Stephanie B.; Verney, Zoe M. [Department of Dermatology, University of Alabama at Birmingham, AL 35294 (United States); Kopelovich, Levy [Division of Cancer Prevention, National Cancer Institute, Bethesda, MD (United States); Elmets, Craig A. [Department of Dermatology, University of Alabama at Birmingham, AL 35294 (United States); Skin Diseases Research Center, University of Alabama at Birmingham, AL 35294 (United States); Athar, Mohammad, E-mail: mathar@uab.edu [Department of Dermatology, University of Alabama at Birmingham, AL 35294 (United States); Skin Diseases Research Center, University of Alabama at Birmingham, AL 35294 (United States)

    2011-05-13

    Research highlights: {yields} Organ transplant recipients are highly susceptible to early skin cancer development. {yields} CsA-mediated TGFB1-dependent TAK1/TAB1 signaling augments invasive tumor growth. {yields} CsA enhances accumulation of upstream kinases, ZMP, AMPK and IRAK to activate TAK1. {yields} TAK1 mediates enhanced proliferation and reduced apoptosis via CsA-dependent NF{kappa}B. -- Abstract: Cyclosporine A (CsA) is an immunosuppressive drug commonly used for maintaining chronic immune suppression in organ transplant recipients. It is known that patients receiving CsA manifest increased growth of aggressive non-melanoma skin cancers. However, the underlying mechanism by which CsA augments tumor growth is not fully understood. Here, we show that CsA augments the growth of A431 epidermoid carcinoma xenograft tumors by activating tumor growth factor {beta}-activated kinase1 (TAK1). The activation of TAK1 by CsA occurs at multiple levels by kinases ZMP, AMPK and IRAK. TAK1 forms heterodimeric complexes with TAK binding protein 1 and 2 (TAB1/TAB2) which in term activate nuclear factor {kappa}B (NF{kappa}B) and p38 MAP kinase. Transcriptional activation of NF{kappa}B is evidenced by IKK{beta}-mediated phosphorylation-dependent degradation of I{kappa}B and consequent nuclear translocation of p65. This also leads to enhancement in the expression of its transcriptional target genes cyclin D1, Bcl2 and COX-2. Similarly, activation of p38 leads to enhanced inflammation-related signaling shown by increased phosphorylation of MAPKAPK2 and which in turn phosphorylates its substrate HSP27. Activation of both NF{kappa}B and p38 MAP kinase provide mitogenic stimuli to augment the growth of SCCs.

  5. TAK1 regulates NF-ΚB and AP-1 activation in airway epithelial cells following RSV infection

    International Nuclear Information System (INIS)

    Dey, Nilay; Liu Tianshuang; Garofalo, Roberto P.; Casola, Antonella

    2011-01-01

    Respiratory syncytial virus (RSV) is the most common cause of epidemic respiratory diseases in infants and young children. RSV infection of airway epithelial cells induces the expression of immune/inflammatory genes through the activation of a subset of transcription factors, including Nuclear Factor-κB (NF-κB) and AP-1. In this study, we have investigated the signaling pathway leading to activation of these two transcription factors in response to RSV infection. Our results show that IKKβ plays a key role in viral-induced NF-κB activation, while JNK regulates AP-1-dependent gene transcription, as demonstrated by using kinase inactive proteins and chemical inhibitors of the two kinases. Inhibition of TAK1 activation, by overexpression of kinase inactive TAK1 or using cells lacking TAK1 expression, significantly reduced RSV-induced NF-κB and AP-1 nuclear translocation and DNA-binding activity, as well as NF-κB-dependent gene expression, identifying TAK1 as an important upstream signaling molecule regulating RSV-induced NF-κB and AP-1 activation. - Highlights: → IKKβ is a major kinase involved in RSV-induced NF-κB activation. → JNK regulates AP-1-dependent gene transcription in RSV infection. → TAK1 is a critical upstream signaling molecule for both pathways in infected cells.

  6. Labdanolic acid methyl ester (LAME) exerts anti-inflammatory effects through inhibition of TAK-1 activation

    International Nuclear Information System (INIS)

    Cuadrado, Irene; Cidre, Florencia; Herranz, Sandra; Estevez-Braun, Ana; Heras, Beatriz de las; Hortelano, Sonsoles

    2012-01-01

    Labdane derivatives obtained from the diterpenoid labdanediol suppressed NO and PGE 2 production in LPS-stimulated RAW 264.7 macrophages. However, mechanisms involved in these inhibitory effects are not elucidated. In this study, we investigated the signaling pathways involved in the anti-inflammatory effects of labdanolic acid methyl ester (LAME) in peritoneal macrophages and examined its therapeutic effect in a mouse endotoxic shock model. LAME reduced the production of NO and PGE 2 in LPS-activated macrophages. This effect involved the inhibition of NOS-2 and COX-2 gene expression, acting at the transcription level. Examination of the effects of the diterpene on NF-κB signaling showed that LAME inhibits the phosphorylation of IκBα and IκBβ, preventing their degradation and the nuclear translocation of the NF-κB p65 subunit. Moreover, inhibition of MAPK signaling was also observed. A further experiment revealed that LAME inhibited the phosphorylation of transforming growth factor-β (TGF-β)-activated kinase 1 (TAK1), an upstream signaling molecule required for IKK and mitogen-activated protein kinases (MAPKs) activation. Inflammatory cytokines such as IL-6, TNF-α and IP-10 were downregulated in the presence of this compound after stimulation with LPS. Additionally, LAME also improved survival in a mouse model of endotoxemia and reduced the circulatory levels of cytokines (IL-6, TNF-α). In conclusion, these results indicate that labdane diterpene LAME significantly attenuates the pro-inflammatory response induced by LPS both in vivo and in vitro. Highlights: ► LAME reduced the production of NO and PGE 2 in LPS-activated macrophages. ► IL-6, TNF-α and IP-10 were also inhibited by LAME. ► Inhibition of TAK-1 activation is the mechanism involved in this process. ► LAME improved survival in a mouse model of endotoxemia. ► LAME reduced the circulatory levels of cytokines (IL-6, TNF-α).

  7. Labdanolic acid methyl ester (LAME) exerts anti-inflammatory effects through inhibition of TAK-1 activation

    Energy Technology Data Exchange (ETDEWEB)

    Cuadrado, Irene [Departamento de Farmacología, Facultad de Farmacia, Universidad Complutense, Plaza Ramón y Cajal s/n, 28040 Madrid (Spain); Cidre, Florencia; Herranz, Sandra [Unidad de Inflamación y Cáncer. Área de Biología Celular y Desarrollo. Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid (Spain); Estevez-Braun, Ana [Instituto Universitario de Bio-Orgánica “Antonio González”. Universidad de La Laguna. Avda. Astrofísico Fco. Sánchez 2. 38206. La Laguna, Tenerife (Spain); Instituto Canario de Investigaciones del Cáncer (ICIC) (Spain); Heras, Beatriz de las, E-mail: lasheras@farm.ucm.es [Departamento de Farmacología, Facultad de Farmacia, Universidad Complutense, Plaza Ramón y Cajal s/n, 28040 Madrid (Spain); Hortelano, Sonsoles, E-mail: shortelano@isciii.es [Unidad de Inflamación y Cáncer. Área de Biología Celular y Desarrollo. Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid (Spain)

    2012-01-01

    Labdane derivatives obtained from the diterpenoid labdanediol suppressed NO and PGE{sub 2} production in LPS-stimulated RAW 264.7 macrophages. However, mechanisms involved in these inhibitory effects are not elucidated. In this study, we investigated the signaling pathways involved in the anti-inflammatory effects of labdanolic acid methyl ester (LAME) in peritoneal macrophages and examined its therapeutic effect in a mouse endotoxic shock model. LAME reduced the production of NO and PGE{sub 2} in LPS-activated macrophages. This effect involved the inhibition of NOS-2 and COX-2 gene expression, acting at the transcription level. Examination of the effects of the diterpene on NF-κB signaling showed that LAME inhibits the phosphorylation of IκBα and IκBβ, preventing their degradation and the nuclear translocation of the NF-κB p65 subunit. Moreover, inhibition of MAPK signaling was also observed. A further experiment revealed that LAME inhibited the phosphorylation of transforming growth factor-β (TGF-β)-activated kinase 1 (TAK1), an upstream signaling molecule required for IKK and mitogen-activated protein kinases (MAPKs) activation. Inflammatory cytokines such as IL-6, TNF-α and IP-10 were downregulated in the presence of this compound after stimulation with LPS. Additionally, LAME also improved survival in a mouse model of endotoxemia and reduced the circulatory levels of cytokines (IL-6, TNF-α). In conclusion, these results indicate that labdane diterpene LAME significantly attenuates the pro-inflammatory response induced by LPS both in vivo and in vitro. Highlights: ► LAME reduced the production of NO and PGE{sub 2} in LPS-activated macrophages. ► IL-6, TNF-α and IP-10 were also inhibited by LAME. ► Inhibition of TAK-1 activation is the mechanism involved in this process. ► LAME improved survival in a mouse model of endotoxemia. ► LAME reduced the circulatory levels of cytokines (IL-6, TNF-α).

  8. Belinostat-induced apoptosis and growth inhibition in pancreatic cancer cells involve activation of TAK1-AMPK signaling axis

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Bing, E-mail: wangbin69@yahoo.com; Wang, Xin-bao; Chen, Li-yu; Huang, Ling; Dong, Rui-zen

    2013-07-19

    Highlights: •Belinostat activates AMPK in cultured pancreatic cancer cells. •Activation of AMPK is important for belinostat-induced cytotoxic effects. •ROS and TAK1 are involved in belinostat-induced AMPK activation. •AMPK activation mediates mTOR inhibition by belinostat. -- Abstract: Pancreatic cancer accounts for more than 250,000 deaths worldwide each year. Recent studies have shown that belinostat, a novel pan histone deacetylases inhibitor (HDACi) induces apoptosis and growth inhibition in pancreatic cancer cells. However, the underlying mechanisms are not fully understood. In the current study, we found that AMP-activated protein kinase (AMPK) activation was required for belinostat-induced apoptosis and anti-proliferation in PANC-1 pancreatic cancer cells. A significant AMPK activation was induced by belinostat in PANC-1 cells. Inhibition of AMPK by RNAi knockdown or dominant negative (DN) mutation significantly inhibited belinostat-induced apoptosis in PANC-1 cells. Reversely, AMPK activator AICAR and A-769662 exerted strong cytotoxicity in PANC-1 cells. Belinostat promoted reactive oxygen species (ROS) production in PANC-1 cells, increased ROS induced transforming growth factor-β-activating kinase 1 (TAK1)/AMPK association to activate AMPK. Meanwhile, anti-oxidants N-Acetyl-Cysteine (NAC) and MnTBAP as well as TAK1 shRNA knockdown suppressed belinostat-induced AMPK activation and PANC-1 cell apoptosis. In conclusion, we propose that belinostat-induced apoptosis and growth inhibition require the activation of ROS-TAK1-AMPK signaling axis in cultured pancreatic cancer cells.

  9. Transforming growth factor beta-activated kinase 1 (TAK1)-dependent checkpoint in the survival of dendritic cells promotes immune homeostasis and function.

    Science.gov (United States)

    Wang, Yanyan; Huang, Gonghua; Vogel, Peter; Neale, Geoffrey; Reizis, Boris; Chi, Hongbo

    2012-02-07

    Homeostatic control of dendritic cell (DC) survival is crucial for adaptive immunity, but the molecular mechanism is not well defined. Moreover, how DCs influence immune homeostasis under steady state remains unclear. Combining DC-specific and -inducible deletion systems, we report that transforming growth factor beta-activated kinase 1 (TAK1) is an essential regulator of DC survival and immune system homeostasis and function. Deficiency of TAK1 in CD11c(+) cells induced markedly elevated apoptosis, leading to the depletion of DC populations, especially the CD8(+) and CD103(+) DC subsets in lymphoid and nonlymphoid tissues, respectively. TAK1 also contributed to DC development by promoting the generation of DC precursors. Prosurvival signals from Toll-like receptors, CD40 and receptor activator of nuclear factor-κB (RANK) are integrated by TAK1 in DCs, which in turn mediated activation of downstream NF-κB and AKT-Foxo pathways and established a gene-expression program. TAK1 deficiency in DCs caused a myeloid proliferative disorder characterized by expansion of neutrophils and inflammatory monocytes, disrupted T-cell homeostasis, and prevented effective T-cell priming and generation of regulatory T cells. Moreover, TAK1 signaling in DCs was required to prevent myeloid proliferation even in the absence of lymphocytes, indicating a previously unappreciated regulatory mechanism of DC-mediated control of myeloid cell-dependent inflammation. Therefore, TAK1 orchestrates a prosurvival checkpoint in DCs that affects the homeostasis and function of the immune system.

  10. Inhibition of transforming growth factor-activated kinase 1 (TAK1 blocks and reverses epithelial to mesenchymal transition of mesothelial cells.

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    Raffaele Strippoli

    Full Text Available Peritoneal fibrosis is a frequent complication of peritoneal dialysis following repeated low grade inflammatory and pro-fibrotic insults. This pathological process may lead to ultrafiltration failure and eventually to the discontinuing of the therapy. Fibrosis is linked to epithelial to mesenchymal transition (EMT of the peritoneal mesothelial cells, which acquire invasive and fibrogenic abilities. Here, we analyzed the role of the transforming growth factor-activated kinase-1 (TAK1 in the EMT of primary mesothelial cells from human peritoneum. The inhibition of TAK1 in mesenchymal-like mesothelial cells from the effluents of patients undergoing peritoneal dialysis led to the reacquisition of the apical to basolateral polarity, to increased expression of epithelial and to down-regulation of mesenchymal markers. TAK1 inhibition also resulted in decreased migratory/invasive abilities of effluent-derived mesothelial cells. Simultaneous inhibition of ERK1/2 and TAK1 pathways did not lead to an additive effect in the reacquisition of the epithelial phenotype. Inhibition of TAK1 also blocked EMT in vitro and reduced the levels of PAI-1, which is involved in fibrosis and invasion. Analysis of signalling pathways downstream of TAK1 involved in EMT induction, showed that TAK1 inhibition reduced the transcriptional activity of NF-κB and Smad3, as well as the phosphorylation of c-jun, while enhancing Smad1-5-8 activity. These results demonstrate that TAK1 is a cross-point in a network including different pro-EMT transcription factors, such as NF-κB, Snail, AP-1 and Smads. The identification of TAK1 as a main biochemical mediator of EMT and fibrosis in mesothelial cells from human peritoneum and the study of signalling pathways induced by its activity may be relevant in the design of new therapies aimed to counteract peritoneal fibrosis.

  11. A Vibrio parahaemolyticus T3SS effector mediates pathogenesis by independently enabling intestinal colonization and inhibiting TAK1 activation.

    Science.gov (United States)

    Zhou, Xiaohui; Gewurz, Benjamin E; Ritchie, Jennifer M; Takasaki, Kaoru; Greenfeld, Hannah; Kieff, Elliott; Davis, Brigid M; Waldor, Matthew K

    2013-05-30

    Vibrio parahaemolyticus type III secretion system 2 (T3SS2) is essential for the organism's virulence, but the effectors required for intestinal colonization and induction of diarrhea by this pathogen have not been identified. Here, we identify a type III secretion system (T3SS2)-secreted effector, VopZ, that is essential for V. parahaemolyticus pathogenicity. VopZ plays distinct, genetically separable roles in enabling intestinal colonization and diarrheagenesis. Truncation of VopZ prevents V. parahaemolyticus colonization, whereas deletion of VopZ amino acids 38-62 abrogates V. parahaemolyticus-induced diarrhea and intestinal pathology but does not impair colonization. VopZ inhibits activation of the kinase TAK1 and thereby prevents the activation of MAPK and NF-κB signaling pathways, which lie downstream. In contrast, the VopZ internal deletion mutant cannot counter the activation of pathways regulated by TAK1. Collectively, our findings suggest that VopZ's inhibition of TAK1 is critical for V. parahaemolyticus to induce diarrhea and intestinal pathology. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

  12. A Vibrio parahaemolyticus T3SS Effector Mediates Pathogenesis by Independently Enabling Intestinal Colonization and Inhibiting TAK1 Activation

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    Xiaohui Zhou

    2013-05-01

    Full Text Available Vibrio parahaemolyticus type III secretion system 2 (T3SS2 is essential for the organism’s virulence, but the effectors required for intestinal colonization and induction of diarrhea by this pathogen have not been identified. Here, we identify a type III secretion system (T3SS2-secreted effector, VopZ, that is essential for V. parahaemolyticus pathogenicity. VopZ plays distinct, genetically separable roles in enabling intestinal colonization and diarrheagenesis. Truncation of VopZ prevents V. parahaemolyticus colonization, whereas deletion of VopZ amino acids 38–62 abrogates V. parahaemolyticus-induced diarrhea and intestinal pathology but does not impair colonization. VopZ inhibits activation of the kinase TAK1 and thereby prevents the activation of MAPK and NF-κB signaling pathways, which lie downstream. In contrast, the VopZ internal deletion mutant cannot counter the activation of pathways regulated by TAK1. Collectively, our findings suggest that VopZ’s inhibition of TAK1 is critical for V. parahaemolyticus to induce diarrhea and intestinal pathology.

  13. Torilin Inhibits Inflammation by Limiting TAK1-Mediated MAP Kinase and NF-κB Activation

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    Mehari Endale

    2017-01-01

    Full Text Available Torilin, a sesquiterpene isolated from the fruits of Torilis japonica, has shown antimicrobial, anticancer, and anti-inflammatory properties. However, data on the mechanism of torilin action against inflammation is limited. This study aimed at determining the anti-inflammatory property of torilin in LPS-induced inflammation using in vitro model of inflammation. We examined torilin’s effect on expression levels of inflammatory mediators and cytokines in LPS-stimulated RAW 264.7 macrophages. The involvement of NF-kB and AP-1, MAP kinases, and adaptor proteins were assessed. Torilin strongly inhibited LPS-induced NO release, iNOS, PGE2, COX-2, NF-α, IL-1β, IL-6, and GM-CSF gene and protein expressions. In addition, MAPKs were also suppressed by torilin pretreatment. Involvement of ERK1/2, P38MAPK, and JNK1/2 was further confirmed by PD98059, SB203580, and SP600125 mediated suppression of iNOS and COX-2 proteins. Furthermore, torilin attenuated NF-kB and AP-1 translocation, DNA binding, and reporter gene transcription. Interestingly, torilin inhibited TAK1 kinase activation with the subsequent suppression of MAPK-mediated JNK, p38, ERK1/2, and AP-1 (ATF-2 and c-jun activation and IKK-mediated I-κBα degradation, p65/p50 activation, and translocation. Together, the results revealed the suppression of NF-κB and AP-1 regulated inflammatory mediator and cytokine expressions, suggesting the test compound’s potential as a candidate anti-inflammatory agent.

  14. TRAF6 promotes myogenic differentiation via the TAK1/p38 mitogen-activated protein kinase and Akt pathways.

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    Fang Xiao

    Full Text Available p38 mitogen-activated protein kinase (MAPK is an essential kinase involved in myogenic differentiation. Although many substrates of p38 MAPK have been identified, little is known about its upstream activators during myogenic differentiation. TRAF6 is known to function in cytokine signaling during inflammatory responses. However, not much is known about its role in myogenic differentiation and muscle regeneration. We showed here that TRAF6 and its intrinsic ubiquitin E3 ligase activity are required for myogenic differentiation. In mouse myoblasts, knockdown of TRAF6 compromised the p38 MAPK and Akt pathways, while deliberate activation of either pathway rescued the differentiation defect caused by TRAF6 knockdown. TAK1 acted as a key signal transducer downstream of TRAF6 in myogenic differentiation. In vivo, knockdown of TRAF6 in mouse muscles compromised the injury-induced muscle regeneration without impairing macrophage infiltration and myoblast proliferation. Collectively, we demonstrated that TRAF6 promotes myogenic differentiation and muscle regeneration via the TAK1/p38 MAPK and Akt pathways.

  15. HTLV-1 Tax Stimulates Ubiquitin E3 Ligase, Ring Finger Protein 8, to Assemble Lysine 63-Linked Polyubiquitin Chains for TAK1 and IKK Activation.

    Science.gov (United States)

    Ho, Yik-Khuan; Zhi, Huijun; Bowlin, Tara; Dorjbal, Batsukh; Philip, Subha; Zahoor, Muhammad Atif; Shih, Hsiu-Ming; Semmes, Oliver John; Schaefer, Brian; Glover, J N Mark; Giam, Chou-Zen

    2015-08-01

    Human T lymphotropic virus type 1 (HTLV-1) trans-activator/oncoprotein, Tax, impacts a multitude of cellular processes, including I-κB kinase (IKK)/NF-κB signaling, DNA damage repair, and mitosis. These activities of Tax have been implicated in the development of adult T-cell leukemia (ATL) in HTLV-1-infected individuals, but the underlying mechanisms remain obscure. IKK and its upstream kinase, TGFβ-activated kinase 1 (TAK1), contain ubiquitin-binding subunits, NEMO and TAB2/3 respectively, which interact with K63-linked polyubiquitin (K63-pUb) chains. Recruitment to K63-pUb allows cross auto-phosphorylation and activation of TAK1 to occur, followed by TAK1-catalyzed IKK phosphorylation and activation. Using cytosolic extracts of HeLa and Jurkat T cells supplemented with purified proteins we have identified ubiquitin E3 ligase, ring finger protein 8 (RNF8), and E2 conjugating enzymes, Ubc13:Uev1A and Ubc13:Uev2, to be the cellular factors utilized by Tax for TAK1 and IKK activation. In vitro, the combination of Tax and RNF8 greatly stimulated TAK1, IKK, IκBα and JNK phosphorylation. In vivo, RNF8 over-expression augmented while RNF8 ablation drastically reduced canonical NF-κB activation by Tax. Activation of the non-canonical NF-κB pathway by Tax, however, is unaffected by the loss of RNF8. Using purified components, we further demonstrated biochemically that Tax greatly stimulated RNF8 and Ubc13:Uev1A/Uev2 to assemble long K63-pUb chains. Finally, co-transfection of Tax with increasing amounts of RNF8 greatly induced K63-pUb assembly in a dose-dependent manner. Thus, Tax targets RNF8 and Ubc13:Uev1A/Uev2 to promote the assembly of K63-pUb chains, which signal the activation of TAK1 and multiple downstream kinases including IKK and JNK. Because of the roles RNF8 and K63-pUb chains play in DNA damage repair and cytokinesis, this mechanism may also explain the genomic instability of HTLV-1-transformed T cells and ATL cells.

  16. TGFβ activated kinase 1 (TAK1 at the crossroad of B cell receptor and Toll-like receptor 9 signaling pathways in human B cells.

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    Dániel Szili

    Full Text Available B cell development and activation are regulated by combined signals mediated by the B cell receptor (BCR, receptors for the B-cell activating factor of the tumor necrosis factor family (BAFF-R and the innate receptor, Toll-like receptor 9 (TLR9. However, the underlying mechanisms by which these signals cooperate in human B cells remain unclear. Our aim was to elucidate the key signaling molecules at the crossroads of BCR, BAFF-R and TLR9 mediated pathways and to follow the functional consequences of costimulation.Therefore we stimulated purified human B cells by combinations of anti-Ig, B-cell activating factor of the tumor necrosis factor family (BAFF and the TLR9 agonist, CpG oligodeoxynucleotide. Phosphorylation status of various signaling molecules, B cell proliferation, cytokine secretion, plasma blast generation and the frequency of IgG producing cells were investigated. We have found that BCR induced signals cooperate with BAFF-R- and TLR9-mediated signals at different levels of cell activation. BCR and BAFF- as well as TLR9 and BAFF-mediated signals cooperate at NFκB activation, while BCR and TLR9 synergistically costimulate mitogen activated protein kinases (MAPKs, ERK, JNK and p38. We show here for the first time that the MAP3K7 (TGF beta activated kinase, TAK1 is responsible for the synergistic costimulation of B cells by BCR and TLR9, resulting in an enhanced cell proliferation, plasma blast generation, cytokine and antibody production. Specific inhibitor of TAK1 as well as knocking down TAK1 by siRNA abrogates the synergistic signals. We conclude that TAK1 is a key regulator of receptor crosstalk between BCR and TLR9, thus plays a critical role in B cell development and activation.

  17. Poxviral protein A52 stimulates p38 mitogen-activated protein kinase (MAPK) activation by causing tumor necrosis factor receptor-associated factor 6 (TRAF6) self-association leading to transforming growth factor β-activated kinase 1 (TAK1) recruitment.

    Science.gov (United States)

    Stack, Julianne; Hurst, Tara P; Flannery, Sinead M; Brennan, Kiva; Rupp, Sebastian; Oda, Shun-ichiro; Khan, Amir R; Bowie, Andrew G

    2013-11-22

    Vaccinia virus encodes a number of proteins that inhibit and manipulate innate immune signaling pathways that also have a role in virulence. These include A52, a protein shown to inhibit IL-1- and Toll-like receptor-stimulated NFκB activation, via interaction with interleukin-1 receptor-associated kinase 2 (IRAK2). Interestingly, A52 was also found to activate p38 MAPK and thus enhance Toll-like receptor-dependent IL-10 induction, which was TRAF6-dependent, but the manner in which A52 manipulates TRAF6 to stimulate p38 activation was unclear. Here, we show that A52 has a non-canonical TRAF6-binding motif that is essential for TRAF6 binding and p38 activation but dispensable for NFκB inhibition and IRAK2 interaction. Wild-type A52, but not a mutant defective in p38 activation and TRAF6 binding (F154A), caused TRAF6 oligomerization and subsequent TRAF6-TAK1 association. The crystal structure of A52 shows that it adopts a Bcl2-like fold and exists as a dimer in solution. Residue Met-65 was identified as being located in the A52 dimer interface, and consistent with that, A52-M65E was impaired in its ability to dimerize. A52-M65E although capable of interacting with TRAF6, was unable to cause either TRAF6 self-association, induce the TRAF6-TAK1 association, or activate p38 MAPK. The results suggest that an A52 dimer causes TRAF6 self-association, leading to TAK1 recruitment and p38 activation. This reveals a molecular mechanism whereby poxviruses manipulate TRAF6 to activate MAPKs (which can be proviral) without stimulating antiviral NFκB activation.

  18. A non-redundant role for Drosophila Mkk4 and hemipterous/Mkk7 in TAK1-mediated activation of JNK.

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    Peter Geuking

    Full Text Available BACKGROUND: The JNK pathway is a mitogen-activated protein (MAP kinase pathway involved in the regulation of numerous physiological processes during development and in response to environmental stress. JNK activity is controlled by two MAPK kinases (MAPKK, Mkk4 and Mkk7. Mkk7 plays a prominent role upon Tumor Necrosis Factor (TNF stimulation. Eiger, the unique TNF-superfamily ligand in Drosophila, potently activates JNK signaling through the activation of the MAPKKK Tak1. METHODOLOGY/PRINCIPAL FINDINGS: In a dominant suppressor screen for new components of the Eiger/JNK-pathway in Drosophila, we have identified an allelic series of the Mkk4 gene. Our genetic and biochemical results demonstrate that Mkk4 is dispensable for normal development and host resistance to systemic bacterial infection but plays a non-redundant role as a MAPKK acting in parallel to Hemipterous/Mkk7 in dTAK1-mediated JNK activation upon Eiger and Imd pathway activation. CONCLUSIONS/SIGNIFICANCE: In contrast to mammals, it seems that in Drosophila both MAPKKs, Hep/Mkk7 and Mkk4, are required to induce JNK upon TNF or pro-inflammatory stimulation.

  19. Tak the hand

    OpenAIRE

    Scott, David D.

    2016-01-01

    In this paper David Scott discusses the theology and rites of ordination of the Scottish Reformed tradition and goes on to reflect on more specific aspects of ministry. His paper takes its title from his examination of the rites of ordination according to Knox’s liturgy whereby, rather than being set apart by the laying on of hands, new ministers were welcomed by colleagues who came forward to ‘tak the hand’ of the ordinand. Following on from a historical survey, the paper includes a critical...

  20. TGF-β-activated kinase 1 (TAK1 signaling regulates TGF-β-induced WNT-5A expression in airway smooth muscle cells via Sp1 and β-catenin.

    Directory of Open Access Journals (Sweden)

    Kuldeep Kumawat

    Full Text Available WNT-5A, a key player in embryonic development and post-natal homeostasis, has been associated with a myriad of pathological conditions including malignant, fibroproliferative and inflammatory disorders. Previously, we have identified WNT-5A as a transcriptional target of TGF-β in airway smooth muscle cells and demonstrated its function as a mediator of airway remodeling. Here, we investigated the molecular mechanisms underlying TGF-β-induced WNT-5A expression. We show that TGF-β-activated kinase 1 (TAK1 is a critical mediator of WNT-5A expression as its pharmacological inhibition or siRNA-mediated silencing reduced TGF-β induction of WNT-5A. Furthermore, we show that TAK1 engages p38 and c-Jun N-terminal kinase (JNK signaling which redundantly participates in WNT-5A induction as only simultaneous, but not individual, inhibition of p38 and JNK suppressed TGF-β-induced WNT-5A expression. Remarkably, we demonstrate a central role of β-catenin in TGF-β-induced WNT-5A expression. Regulated by TAK1, β-catenin is required for WNT-5A induction as its silencing repressed WNT-5A expression whereas a constitutively active mutant augmented basal WNT-5A abundance. Furthermore, we identify Sp1 as the transcription factor for WNT-5A and demonstrate its interaction with β-catenin. We discover that Sp1 is recruited to the WNT-5A promoter in a TGF-β-induced and TAK1-regulated manner. Collectively, our findings describe a TAK1-dependent, β-catenin- and Sp1-mediated signaling cascade activated downstream of TGF-β which regulates WNT-5A induction.

  1. Norisoboldine suppresses osteoclast differentiation through preventing the accumulation of TRAF6-TAK1 complexes and activation of MAPKs/NF-κB/c-Fos/NFATc1 Pathways.

    Directory of Open Access Journals (Sweden)

    Zhi-Feng Wei

    Full Text Available Norisoboldine (NOR is the main alkaloid constituent in the dry root of Lindera aggregata (Sims Kosterm. (L. strychnifolia Vill.. As reported previously, orally administered NOR displayed a robust inhibition of joint bone destruction present in both mouse collagen-induced arthritis and rat adjuvant-induced arthritis with lower efficacious doses than that required for ameliorating systemic inflammation. This attracted us to assess the effects of NOR on differentiation and function of osteoclasts, primary effector cells for inflammatory bone destruction, to get insight into its anti-rheumatoid arthritis mechanisms. Both RAW264.7 cells and mouse bone marrow-derived macrophages (BMMs were stimulated with RANKL (100 ng/mL to establish osteoclast differentiation models. ELISA, RT-PCR, gelatin zymography, western blotting, immunoprecipitation and EMSA were used to reveal related signalling pathways. NOR (10 and 30 µM, without significant cytotoxicity, showed significant reduction of the number of osteoclasts and the resorption pit areas, and it targeted osteoclast differentiation at the early stage. In conjunction with the anti-resorption effect of NOR, mRNA levels of cathepsin K and MMP-9 were decreased, and the activity of MMP-9 was attenuated. Furthermore, our mechanistic studies indicated that NOR obviously suppressed the ubiquitination of TRAF6, the accumulation of TRAF6-TAK1 complexes and the activation of ERK and p38 MAPK, and reduced the nuclear translocation of NF-κB-p65 and DNA-binding activity of NF-κB. However, NOR had little effect on expressions of TRAF6 or the phosphorylation and degradation of IκBα. Moreover, NOR markedly inhibited expressions of transcription factor NFATc1, but not c-Fos. Intriguingly, the subsequent nuclear translocations of c-Fos and NFATc1 were substantially down-regulated. Hence, we demonstrated for the first time that preventing the differentiation and function of osteoclasts at the early stage was an

  2. TAK1 modulates satellite stem cell homeostasis and skeletal muscle repair

    Science.gov (United States)

    Ogura, Yuji; Hindi, Sajedah M.; Sato, Shuichi; Xiong, Guangyan; Akira, Shizuo; Kumar, Ashok

    2015-01-01

    Satellite cells are resident adult stem cells that are required for regeneration of skeletal muscle. However, signalling mechanisms that regulate satellite cell function are less understood. Here we demonstrate that transforming growth factor-β-activated kinase 1 (TAK1) is important in satellite stem cell homeostasis and function. Inactivation of TAK1 in satellite cells inhibits muscle regeneration in adult mice. TAK1 is essential for satellite cell proliferation and its inactivation causes precocious differentiation. Moreover, TAK1-deficient satellite cells exhibit increased oxidative stress and undergo spontaneous cell death, primarily through necroptosis. TAK1 is required for the activation of NF-κB and JNK in satellite cells. Forced activation of NF-κB improves survival and proliferation of TAK1-deficient satellite cells. Furthermore, TAK1-mediated activation of JNK is essential to prevent oxidative stress and precocious differentiation of satellite cells. Collectively, our study suggests that TAK1 is required for maintaining the pool of satellite stem cells and for regenerative myogenesis. PMID:26648529

  3. TAK1 binding protein 2 is essential for liver protection from stressors.

    Directory of Open Access Journals (Sweden)

    Yuka Ikeda

    Full Text Available The liver is the first line of defense from environmental stressors in that hepatocytes respond to and metabolize them. Hence, hepatocytes can be damaged by stressors. Protection against hepatic cell damage and cell death is important for liver function and homeostasis. TAK1 (MAP3K7 is an intermediate of stressors such as bacterial moieties-induced signal transduction pathways in several cell types. Tak1 deficiency has been reported to induce spontaneous hepatocellular carcinoma. However, the regulatory mechanism of TAK1 activity in liver stress response has not yet been defined. Here we report that activation of TAK1 through TAK1 binding protein 2 (TAB2 is required for liver protection from stressors. We found that a bacterial moiety, lipopolysaccharides (LPS, activated TAK1 in primary hepatocytes, which was diminished by deletion of TAB2. Mice having hepatocyte-specific deletion of the Tab2 gene exhibited only late-onset moderate liver lesions but were hypersensitive to LPS-induced liver injury. Furthermore, we show that a chemical stressor induced greatly exaggerated liver injury in hepatocyte-specific Tab2-deficient mice. These results demonstrate that TAB2 is a sensor of stress conditions in the liver and functions to protect the liver by activating the TAK1 pathway.

  4. Adenovirus-Mediated Small Interfering RNA Targeting TAK1 Ameliorates Joint Inflammation with Collagen-Induced Arthritis in Mice.

    Science.gov (United States)

    Luo, Xinjing; Chen, Yongfeng; Lv, Guoju; Zhou, Zhidong; Chen, Jie; Mo, Xuanrong; Xie, Jiangwen

    2017-06-01

    Transforming growth factor β-activated kinase-1 (TAK1) is a key upstream kinase in cell signaling during inflammation, which regulates the expression of inflammatory mediators. Small interfering RNA (siRNA) against TAK1 offers promise as a potential therapeutic strategy in immune-mediated inflammatory disorder including rheumatoid arthritis. Here, we are to evaluate the therapeutic effects of intra-articular administration of adenoviral-mediated siRNA against TAK1 (ad-siRNA-TAK1) on collagen-induced arthritis (CIA) in mice. Ad-siRNA-TAK1 was constructed. The murine RAW 264.7 macrophages were infected with ad-siRNA-TAK1, and the silencing specificity of TAK1 was assessed by quantitative polymerase chain reaction (PCR) and western blot. DBA/1 mice were injected intra-articularly with ad-siRNA-TAK1. Development and severity of arthritis was assessed histologically. Synovial inflammation and bone destruction were determined by hematoxylin and eosin (HE) staining. Articular and serum concentrations of tumor necrosis factor-α, interleukin-1, and interleukin-6 were determined using enzyme-linked immunosorbent assay. Levels of phosphorylated p38, c-Jun N-terminal kinase (JNK), and extracellular-signal-regulated kinase (ERK) were detected by western blot. In vitro, ad--siRNA-TAK1 efficiently inhibited the expression of TAK1 at both mRNA and protein levels. In vivo, intra-articular injection of ad-siRNA-TAK1 efficiently alleviated joint inflammation, decreased the expression of pro-inflammatory mediators, and suppressed JNK pathways. Our results demonstrate the efficiency of ad--siRNA-TAK1 in controlling joint inflammation of CIA, which is associated with the suppression of the expression of pro-inflammatory cytokines and JNK activation.

  5. Some problems of residual activity measurements

    International Nuclear Information System (INIS)

    Katrik, P.; Mustafin, E.; Strasik, I.; Pavlovic, M.

    2013-01-01

    As a preparatory work for constructing the Facility for Antiproton and Ion Research (FAIR) at GSI Darmstadt, samples of copper were irradiated by 500 MeV/u 238 U ion beam and investigated by gamma-ray spectroscopy. The nuclides that contribute dominantly to the residual activity have been identified and their contributions have been quantified by two different methods: from the whole-target gamma spectra and by integration of depth-profiles of residual activity of individual nuclides. Results obtained by these two methods are compared and discussed in this paper. (authors)

  6. Kepedulian Lingkungan Dalam Jamangilak Tak Pernah Menangis

    Directory of Open Access Journals (Sweden)

    Lustantini Septiningsih

    2015-06-01

    Full Text Available Penelitian ini bertujuan mendeskripsikan kepedulian lingkungan dalam novel Jamangilak Tak Pernah Menangis karya Martin Aleida. Metode yang digunakan dalam penelitian ini adalah metode deskriptif, yaitu penelitian dilakukan atas dasar fakta yang ada sehingga pemerian yang dikemukakan sesuai dengan fakta. Untuk mengetahui kepedulian terhadap lingkungan yang terdapat dalam novel Jamangilak Tak Pernah Menangis digunakan pendekatan sosiologi sastra, yaitu bentuk penelaahan sastra yang mempertimbangkan unsur sosial atau kemasyarakatan. Hasil penelitian ini menunjukkan bahwa kepedulian yang dilakukan oleh tokoh Molek meliputi kepedulian terhadap sungai, kepedulian terhadap Danau Toba, dan kepedulian terhadap moral. Kepedulian Molek tersebut didasari oleh tidak adanya tindakan pemerintah dalam melihat daerah yang mengalami pendangkalan sungai serta adanya polusi air dan udara dari limbah pabrik bubur kayu. Kepeduliannya terhadap moral didasari oleh tanggung jawab Molek sebagai ibu yang selalu menanamkan ajaran agama dan nilai kehidupan yang berlaku dalam masyarakat dalam mendidik anaknya. Abstract: This research aims to describe environmental awareness in novel Jamangilak Tak Pernah Menangis by Martin Aleida. The method used in this research is descriptive method; the research was conducted based on fact so that the description presented was consistent with the fact. To identify the awareness on environmental issues in Jamangilak Tak Pernah Menangis, the researcher used the sociology of literature approach which is a form of literary study considering society factor. The result of this research shows careness which conducted by Molek’s figure includes careness of river, careness of lake Toba, and careness of moral. Those careness based on none of government’s action to views the area affected by river depth of air and water pollution from the industrial wood pulp. The careness of moral based on her responsibility as a mother who always instilled

  7. Macropinocytosis and TAK1 mediate anti-inflammatory to pro-inflammatory macrophage differentiation by HIV-1 Nef.

    Science.gov (United States)

    Hashimoto, M; Nasser, H; Chihara, T; Suzu, S

    2014-05-29

    Macrophages (MΦ) are functionally classified into two types, anti-inflammatory M2 and pro-inflammatory M1. Importantly, we recently revealed that soluble HIV-1 proteins, particularly the pathogenetic protein Nef, preferentially activate M2-MΦ and drive them towards an M1-like MΦ, which might explain the sustained immune activation seen in HIV-1-infected patients. Here, we show that the preferential effect of Nef on M2-MΦ is mediated by TAK1 (TGF-β-activated kinase 1) and macropinocytosis. As with MAP kinases and NF-κB pathway, Nef markedly activated TAK1 in M-CSF-derived M2-MΦ but not in GM-CSF-derived M1-MΦ. Two Nef mutants, which were unable to activate MAP kinases and NF-κB pathway, failed to activate TAK1. Indeed, the TAK1 inhibitor 5Z-7-oxozeaenol as well as the ectopic expression of a dominant-negative mutant of TAK1 or TRAF2, an upstream molecule of TAK1, inhibited Nef-induced signaling activation and M1-like phenotypic differentiation of M2-MΦ. Meanwhile, the preferential effect of Nef on M2-MΦ correlated with the fact the Nef entered M2-MΦ more efficiently than M1-MΦ. Importantly, the macropinosome formation inhibitor EIPA completely blocked the internalization of Nef into M2-MΦ. Because the macropinocytosis activity of M2-MΦ was higher than that of M1-MΦ, our findings indicate that Nef enters M2-MΦ efficiently by exploiting their higher macropinocytosis activity and drives them towards M1-like MΦ by activating TAK1.

  8. Characterization of binding and inhibitory properties of TAK-063, a novel phosphodiesterase 10A inhibitor.

    Directory of Open Access Journals (Sweden)

    Akina Harada

    Full Text Available Phosphodiesterase 10A (PDE10A inhibition is a novel and promising approach for the treatment of central nervous system disorders such as schizophrenia and Huntington's disease. A novel PDE10A inhibitor, TAK-063 [1-[2-fluoro-4-(1H-pyrazol-1-ylphenyl]-5-methoxy-3-(1-phenyl-1H-pyrazol-5-yl-pyridazin-4(1H-one] has shown high inhibitory activity and selectivity for human recombinant PDE10A2 in vitro; the half-maximal inhibitory concentration was 0.30 nM, and selectivity over other phosphodiesterases (PDEs was more than 15000-fold. TAK-063 at 10 µM did not show more than 50% inhibition or stimulation of 91 enzymes or receptors except for PDEs. In vitro autoradiography (ARG studies using rat brain sections revealed that [3H]TAK-063 selectively accumulated in the caudate putamen (CPu, nucleus accumbens (NAc, globus pallidus, substantia nigra, and striatonigral projection, where PDE10A is highly expressed. This [3H]TAK-063 accumulation was almost entirely blocked by an excess amount of MP-10, a PDE10A selective inhibitor, and the accumulation was not observed in brain slices of Pde10a-knockout mice. In rat brain sections, [3H]TAK-063 bound to a single high-affinity site with mean ± SEM dissociation constants of 7.2 ± 1.2 and 2.6 ± 0.5 nM for the CPu and NAc shell, respectively. Orally administered [14C]TAK-063 selectively accumulated in PDE10A expressing brain regions in an in vivo ARG study in rats. Striatal PDE10A occupancy by TAK-063 in vivo was measured using T-773 as a tracer and a dose of 0.88 mg/kg (p.o. was calculated to produce 50% occupancy in rats. Translational studies with TAK-063 and other PDE10A inhibitors such as those presented here will help us better understand the pharmacological profile of this class of potential central nervous system drugs.

  9. Mice with Tak1 Deficiency in Neural Crest Lineage Exhibit Cleft Palate Associated with Abnormal Tongue Development*

    Science.gov (United States)

    Song, Zhongchen; Liu, Chao; Iwata, Junichi; Gu, Shuping; Suzuki, Akiko; Sun, Cheng; He, Wei; Shu, Rong; Li, Lu; Chai, Yang; Chen, YiPing

    2013-01-01

    Cleft palate represents one of the most common congenital birth defects in humans. TGFβ signaling, which is mediated by Smad-dependent and Smad-independent pathways, plays a crucial role in regulating craniofacial development and patterning, particularly in palate development. However, it remains largely unknown whether the Smad-independent pathway contributes to TGFβ signaling function during palatogenesis. In this study, we investigated the function of TGFβ activated kinase 1 (Tak1), a key regulator of Smad-independent TGFβ signaling in palate development. We show that Tak1 protein is expressed in both the epithelium and mesenchyme of the developing palatal shelves. Whereas deletion of Tak1 in the palatal epithelium or mesenchyme did not give rise to a cleft palate defect, inactivation of Tak1 in the neural crest lineage using the Wnt1-Cre transgenic allele resulted in failed palate elevation and subsequently the cleft palate formation. The failure in palate elevation in Wnt1-Cre;Tak1F/F mice results from a malformed tongue and micrognathia, resembling human Pierre Robin sequence cleft of the secondary palate. We found that the abnormal tongue development is associated with Fgf10 overexpression in the neural crest-derived tongue tissue. The failed palate elevation and cleft palate were recapitulated in an Fgf10-overexpressing mouse model. The repressive effect of the Tak1-mediated noncanonical TGFβ signaling on Fgf10 expression was further confirmed by inhibition of p38, a downstream kinase of Tak1, in the primary cell culture of developing tongue. Tak1 thus functions to regulate tongue development by controlling Fgf10 expression and could represent a candidate gene for mutation in human PRS clefting. PMID:23460641

  10. Lack of TAK1 in dendritic cells inhibits the contact hypersensitivity response induced by trichloroethylene in local lymph node assay

    International Nuclear Information System (INIS)

    Yao, Pan; Hongqian, Chu; Qinghe, Meng; Lanqin, Shang; Jianjun, Jiang; Xiaohua, Yang; Xuetao, Wei; Weidong, Hao

    2016-01-01

    Trichloroethylene (TCE) is a ubiquitous environmental contaminant. Occupational TCE exposure has been associated with severe, generalized contact hypersensitivity (CHS) skin disorder. The development of CHS depends on innate and adaptive immune functions. Transforming growth factor-β activated kinase-1 (TAK1) controls the survival of dendritic cells (DCs) that affect the immune system homeostasis. We aimed to investigate the role of TAK1 activity in DC on TCE-induced CHS response. Control mice and DC-specific TAK1 deletion mice were treated with 80% (v/v) TCE using local lymph node assay (LLNA) to establish a TCE-induced CHS model. The draining lymph nodes (DLNs) were excised and the lymphocytes were measure for proliferation by BrdU-ELISA, T-cell phenotype analysis by flow cytometry and signaling pathway activation by western blot. The ears were harvested for histopathological analysis. Control mice in the 80% TCE group displayed an inflammatory response in the ears, increased lymphocyte proliferation, elevated regulatory T-cell and activated T-cell percentages, and more IFN-γ producing CD8 + T cells in DLNs. In contrast to control mice, DC-specific TAK1 deletion mice in the 80% TCE group showed an abolished CHS response and this was associated with defective T-cell expansion, activation and IFN-γ production. This effect may occur through Jnk and NF-κB signaling pathways. Overall, this study demonstrates a pivotal role of TAK1 in DCs in controlling TCE-induced CHS response and suggests that targeting TAK1 function in DCs may be a viable approach to preventing and treating TCE-related occupational health hazards. - Highlights: • Lack of TAK1 in DC caused an abolished TCE-induced CHS response. • TAK1 in DCs was essential to maintain the homeostasis of T cells in TCE-induced CHS. • Intact TAK1 in DCs was critical to promote T-cell priming in TCE-induced CHS. • DC-specific TAK1 deficiency abolished the TCE-mediated phosphorylation of Jnk.

  11. Alkali activation processes for incinerator residues management.

    Science.gov (United States)

    Lancellotti, Isabella; Ponzoni, Chiara; Barbieri, Luisa; Leonelli, Cristina

    2013-08-01

    Incinerator bottom ash (BA) is produced in large amount worldwide and in Italy, where 5.1 millionstons of municipal solid residues have been incinerated in 2010, corresponding to 1.2-1.5 millionstons of produced bottom ash. This residue has been used in the present study for producing dense geopolymers containing high percentage (50-70 wt%) of ash. The amount of potentially reactive aluminosilicate fraction in the ash has been determined by means of test in NaOH. The final properties of geopolymers prepared with or without taking into account this reactive fraction have been compared. The results showed that due to the presence of both amorphous and crystalline fractions with a different degree of reactivity, the incinerator BA geopolymers exhibit significant differences in terms of Si/Al ratio and microstructure when reactive fraction is considered. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. IL-17F induces IL-6 via TAK1-NFκB pathway in airway smooth muscle cells.

    Science.gov (United States)

    Nakajima, Masayuki; Kawaguchi, Mio; Ota, Kyoko; Fujita, Junichi; Matsukura, Satoshi; Huang, Shau-Ku; Morishima, Yuko; Ishii, Yukio; Satoh, Hiroaki; Sakamoto, Tohru; Hizawa, Nobuyuki

    2017-06-01

    Interleukin (IL)-17F plays a critical role in the pathophysiology of asthma. However, the precise role of IL-17F in airway smooth muscle cells (ASMCs) and its regulatory mechanisms remain to be defined. Therefore, we sought to investigate the expression of IL-6 by IL-17F and the involvement of transforming growth factor β-activated kinase 1 (TAK1) and nuclear factor (NF)-κB by in ASMCs. ASMCs were cultured in the presence or absence of IL-17F. The expression of IL-6 gene and protein was analyzed using real-time PCR and ELISA, and the activation of TAK1 and NF-κB was detected by Western blotting. The effect of TAK1 inhibitor 5Z-7-oxozeaenol and NF-κB inhibitor BAY 11-7082 on the expression of IL-6 was investigated. Finally, the short interfering RNAs (siRNAs) targeting TAK1 and a subunit of NF-κB, p65 were transfected into ASMCs. The expression of IL-6 gene and protein was significantly induced by IL-17F. IL-17F activated TAK1 and NF-κB in ASMCs. Transfection of siRNAs targeting TAK1 abolished IL-17F-induced phosphorylation of p65. Both 5Z-7-oxozeaenol and BAY 11-7082 significantly inhibited IL-17F-induced IL-6 production in a dose-dependent manner. Similarly, transfection of the cells with siRNAs targeting TAK1 and p65 inhibited the expression of IL-6. Collectively, these results provided evidence supporting the potential importance of the Th17-ASMCs crosstalk via the IL-17F-IL-6 axis in airway inflammation and as a candidate pharmacological target for airway inflammatory diseases such as asthma. © 2017 The Authors. Immunity, Inflammation and Disease Published by John Wiley & Sons Ltd.

  13. IL‐17F induces IL‐6 via TAK1‐NFκB pathway in airway smooth muscle cells

    Science.gov (United States)

    Nakajima, Masayuki; Ota, Kyoko; Fujita, Junichi; Matsukura, Satoshi; Huang, Shau‐Ku; Morishima, Yuko; Ishii, Yukio; Satoh, Hiroaki; Sakamoto, Tohru; Hizawa, Nobuyuki

    2017-01-01

    Abstract Introduction Interleukin (IL)‐17F plays a critical role in the pathophysiology of asthma. However, the precise role of IL‐17F in airway smooth muscle cells (ASMCs) and its regulatory mechanisms remain to be defined. Therefore, we sought to investigate the expression of IL‐6 by IL‐17F and the involvement of transforming growth factor β‐activated kinase 1 (TAK1) and nuclear factor (NF)‐κB by in ASMCs. Methods ASMCs were cultured in the presence or absence of IL‐17F. The expression of IL‐6 gene and protein was analyzed using real‐time PCR and ELISA, and the activation of TAK1 and NF‐κB was detected by Western blotting. The effect of TAK1 inhibitor 5Z‐7‐oxozeaenol and NF‐κB inhibitor BAY 11‐7082 on the expression of IL‐6 was investigated. Finally, the short interfering RNAs (siRNAs) targeting TAK1 and a subunit of NF‐κB, p65 were transfected into ASMCs. Results The expression of IL‐6 gene and protein was significantly induced by IL‐17F. IL‐17F activated TAK1 and NF‐κB in ASMCs. Transfection of siRNAs targeting TAK1 abolished IL‐17F‐induced phosphorylation of p65. Both 5Z‐7‐oxozeaenol and BAY 11‐7082 significantly inhibited IL‐17F‐induced IL‐6 production in a dose‐dependent manner. Similarly, transfection of the cells with siRNAs targeting TAK1 and p65 inhibited the expression of IL‐6. Conclusions Collectively, these results provided evidence supporting the potential importance of the Th17‐ASMCs crosstalk via the IL‐17F‐IL‐6 axis in airway inflammation and as a candidate pharmacological target for airway inflammatory diseases such as asthma. PMID:28474507

  14. Secondary polycythaemia in a Malay girl with homozygous Hb Tak.

    Science.gov (United States)

    Amran, H S; Aziz, M A; George, E; Mahmud, N; Lee, T Y; Md Noor, S

    2017-12-01

    Hb Tak is one of more than 200 high affinity haemoglobin variants reported worldwide. It results from the insertion of two nucleotides (AC) at the termination codon, between codon 146 and codon 147 of the beta-globin gene [Beta 147 (+AC)]. Polycythaemia is the main clinical feature although affected carriers are usually asymptomatic and do not require intervention. Several case studies in this region have reported the co-inheritance of Hb Tak with Hb E, delta beta and beta thalassaemia with one case of homozygous Hb Tak in a Thai boy. In this case report, a cluster of haemoglobin Tak was found in a family of Malay ethnic origin. Cascade family screening was conducted while investigating a 4-year old girl who presented with symptomatic polycythaemia. She had 2 previous Hb analysis done, at 7-month and 2-year-old with the diagnosis of possible Hb Q Thailand and Homozygous Hb D, respectively. Both diagnosis did not fit her clinical presentations. She was plethoric, had reduced exercise tolerance as well as cardiomyopathy. Her parents were consanguineously married and later diagnosed as asymptomatic carriers of Hb Tak. Consequently, re-analysis of the girl's blood sample revealed a homozygous state of Hb Tak. In conclusion, high oxygen affinity haemoglobin like Hb Tak should be considered in the investigation of polycythaemic patients with abnormal Hb analyses. In this case, DNA analysis was crucial in determining the correct diagnosis.

  15. Suppressor of cytokine Signaling-3 inhibits interleukin-1 signaling by targeting the TRAF-6/TAK1 complex

    DEFF Research Database (Denmark)

    Frobøse, Helle; Rønn, Sif Groth; Heding, Peter E

    2006-01-01

    IL-1 plays a major role in inflammation and autoimmunity through activation of nuclear factor kappa B (NFkappaB) and MAPKs. Although a great deal is known about the mechanism of activation of NFkappaB and MAPKs by IL-1, much less is known about the down-regulation of this pathway. Suppressor......-activated kinase 1, TNF receptor-associated factor (TRAF)6, and TGFbeta-activated kinase (TAK)1, but not when the MAP3K MAPK/ERK kinase kinase-1 is used instead of TAK1, indicating that the target for SOCS-3 is the TRAF6/TAK1 signaling complex. By coimmunoprecipitation, it was shown that SOCS-3 inhibited...

  16. Transcriptional corepressors HIPK1 and HIPK2 control angiogenesis via TGF-β-TAK1-dependent mechanism.

    Directory of Open Access Journals (Sweden)

    Yulei Shang

    Full Text Available Several critical events dictate the successful establishment of nascent vasculature in yolk sac and in the developing embryos. These include aggregation of angioblasts to form the primitive vascular plexus, followed by the proliferation, differentiation, migration, and coalescence of endothelial cells. Although transforming growth factor-β (TGF-β is known to regulate various aspects of vascular development, the signaling mechanism of TGF-β remains unclear. Here we show that homeodomain interacting protein kinases, HIPK1 and HIPK2, are transcriptional corepressors that regulate TGF-β-dependent angiogenesis during embryonic development. Loss of HIPK1 and HIPK2 leads to marked up-regulations of several potent angiogenic genes, including Mmp10 and Vegf, which result in excessive endothelial proliferation and poor adherens junction formation. This robust phenotype can be recapitulated by siRNA knockdown of Hipk1 and Hipk2 in human umbilical vein endothelial cells, as well as in endothelial cell-specific TGF-β type II receptor (TβRII conditional mutants. The effects of HIPK proteins are mediated through its interaction with MEF2C, and this interaction can be further enhanced by TGF-β in a TAK1-dependent manner. Remarkably, TGF-β-TAK1 signaling activates HIPK2 by phosphorylating a highly conserved tyrosine residue Y-361 within the kinase domain. Point mutation in this tyrosine completely eliminates the effect of HIPK2 as a transcriptional corepressor in luciferase assays. Our results reveal a previously unrecognized role of HIPK proteins in connecting TGF-β signaling pathway with the transcriptional programs critical for angiogenesis in early embryonic development.

  17. Studi Sistem Akustik Pada Gereja Katolik Santa Maria Tak Bercela Surabaya

    OpenAIRE

    Hedy C. Indrani, Lisayana

    2013-01-01

    The church is a building that has an image of God's glory so that through space design elements, sacred atmosphere can formone through acoustics. The church has an unique acoustic because the church has two activities, namely speech and music. Santa Maria Tak Bercela Catholic Church Surabaya adjacent to Santa Clara School Surabaya, which at certain hours of the atmosphere will be very crowded church. With the help of Sound Level Meter, the background noise is known and the power of the sound ...

  18. Antioxidant activity of extracts obtained from residues of different oilseeds.

    Science.gov (United States)

    Matthäus, B

    2002-06-05

    Residues of the oil-extracting process of oilseeds contain bioactive substances such as phenolic compounds, which could be used as natural antioxidants for the protection of fats and oils against oxidative deterioration. Thus, the extraction of such constituents from residual material can be considered to contribute to the added value of these residues, which could justify their isolation. In the present work the fat-free residues of eight different oilseeds whose oils are usable for nutritional applications, and also as renewable resources, were extracted with 70% methanol, 70% acetone, water, and ethyl acetate/water, respectively. The resulting extracts were investigated regarding their content of total phenolic compounds by the Folin-Ciocalteau assay, sinapine, flavanoids, and the UV-absorption spectra. Further, the antioxidant activity of the extracts was characterized by the DPPH method, the beta-carotene-linoleic acid assay, and ESR investigations. The fat-free residues of the different oilseeds contained considerable amounts of extractable substances. The yields decreased with decreasing polarity of the solvent in the order water, 70% methanol, 70% acetone, and ethyl acetate. The ratio of total phenolic compounds to the extractable compounds ranged from 3 to 19%. There was no significant correlation between the amount of total extractable compounds and the total phenolic compounds (p < 0.001). All extracts showed remarkable antioxidant activities determined with the different methods. The effects depended strongly on the solvent used for the extraction as well as on the extracted residue. A correlation between the methods used for the characterization of the antioxidant activity and the composition of the residues could not be shown.

  19. Antihypertensive Activity of Residue From “Gebto Arekei”. Locally ...

    African Journals Online (AJOL)

    Antihypertensive Activity of Residue From “Gebto Arekei”. Locally Distilled Medicinal Spirit From a Brew Containing Lupinus albus Seeds in Renovascular Hypertensive Guines-Pigs. Cherinet Ambaye, Tesfaye Tolessa, Abebe Abera, Hassen Taha Sherief, Dawit Abebe, Kelbessa Urga ...

  20. European standardization activities on residual stress analysis by neutron diffraction

    CERN Document Server

    Youtsos, A G

    2002-01-01

    A main objective of a recently completed European research project, RESTAND - residual stress standard using neutron diffraction, was to develop industrial confidence in the application of the neutron-diffraction technique for residual stress measurement and its principal deliverable was a relevant draft code of practice. In fact this draft standard was jointly developed within RESTAND and VAMAS TWA 20 - an international pre-normative research activity. As no such standard is yet available, on the basis of this draft standard document the European Standards Committee on Non-Destructive Testing (CEN TC/138) has established a new ad hoc Work Group (AHG7). The objective of this group is the development of a European pre-standard on a 'test method for measurement of residual stress by neutron diffraction'. The document contains the proposed protocol for making the measurements. It includes the scope of the method, an outline of the technique, the calibration and measurement procedures recommended, and details of ...

  1. Neutron activation analysis for noble metals in matte leach residues

    International Nuclear Information System (INIS)

    Hart, R.J.

    1978-01-01

    The development of the neutron activation analysis technique as a method for rapid and precise determinations of platinum group metals in matte leach residues depends on obtaining a method for effecting complete and homogeneous sample dilution. A simple method for solid dilution of metal samples is outlined in this study, which provided a basis for the accurate determination of all the noble metals by the Neutron Activation Analysis technique

  2. Synergistic effect of a factor Xa inhibitor, TAK-442, and antiplatelet agents on whole blood coagulation and arterial thrombosis in rats.

    Science.gov (United States)

    Konishi, Noriko; Hiroe, Katsuhiko; Kawamura, Masaki

    2010-08-01

    Activated platelets facilitate blood coagulation by providing factor V and a procoagulant surface for prothrombinase. Here, we investigated the potential synergy of a potent factor Xa/prothrombinase inhibitor, TAK-442, plus aspirin or clopidogrel in preventing arterial thrombosis and whole blood coagulation. Thrombus formation was initiated by FeCl(3)-induced rat carotid injury. Bleeding time was evaluated with the rat tail transection model. Whole blood coagulation was assessed by thromboelastographic examination (TEG) for which blood obtained from control, aspirin-, or clopidogrel-treated rats was transferred to a TEG analyzer containing, collagen or adenosine diphosphate (ADP), and TAK-442 or vehicle. TAK-442 (3mg/kg, po), aspirin (100mg/kg, po) or clopidogrel (3mg/kg, po) alone had no significant effect on thrombus formation, whereas the combination of TAK-442 with aspirin and clopidogrel remarkably prolonged the time to thrombus formation without additional significant prolongation of bleeding time. TEG demonstrated that the onset of collagen-induced blood coagulation were slightly longer in aspirin-treated rats than control; however, when the blood from aspirin-treated rats was subsequently treated in vitro with 100 nM TAK-442, the onset of clotting was significantly prolonged. In contrast, only marginal prolongation was observed with TAK-442 treatment of blood from control animals. The onset time of ADP-induced blood coagulation was slightly longer in clopidogrel-treated rats compared with control, and it was further extended by TAK-442 treatment. These results demonstrate that blood coagulation can be markedly delayed by the addition of TAK-442 to antiplatelets treatment which could contribute to synergistic antithrombotic efficacy in these settings. (c) 2010 Elsevier Ltd. All rights reserved.

  3. Prediction of Active Site and Distal Residues in E. coli DNA Polymerase III alpha Polymerase Activity.

    Science.gov (United States)

    Parasuram, Ramya; Coulther, Timothy A; Hollander, Judith M; Keston-Smith, Elise; Ondrechen, Mary Jo; Beuning, Penny J

    2018-02-20

    The process of DNA replication is carried out with high efficiency and accuracy by DNA polymerases. The replicative polymerase in E. coli is DNA Pol III, which is a complex of 10 different subunits that coordinates simultaneous replication on the leading and lagging strands. The 1160-residue Pol III alpha subunit is responsible for the polymerase activity and copies DNA accurately, making one error per 10 5 nucleotide incorporations. The goal of this research is to determine the residues that contribute to the activity of the polymerase subunit. Homology modeling and the computational methods of THEMATICS and POOL were used to predict functionally important amino acid residues through their computed chemical properties. Site-directed mutagenesis and biochemical assays were used to validate these predictions. Primer extension, steady-state single-nucleotide incorporation kinetics, and thermal denaturation assays were performed to understand the contribution of these residues to the function of the polymerase. This work shows that the top 15 residues predicted by POOL, a set that includes the three previously known catalytic aspartate residues, seven remote residues, plus five previously unexplored first-layer residues, are important for function. Six previously unidentified residues, R362, D405, K553, Y686, E688, and H760, are each essential to Pol III activity; three additional residues, Y340, R390, and K758, play important roles in activity.

  4. Exploring a model of human chemokine receptor CCR2 in presence of TAK779: A membrane based molecular dynamics study

    Science.gov (United States)

    Balupuri, Anand; Sobhia, M. Elizabeth

    2014-04-01

    Chemokine receptor 2 (CCR2) is a G-protein coupled receptor (GPCR) and a crucial target for various inflammation-driven diseases. In the present study, molecular docking and molecular dynamics simulations were performed on a CCR2 homology model. This work includes the comparative MD simulations of uncomplexed and ‘antagonist-complexed’ CCR2 models. These simulations yield insights into the binding mechanism of antagonist TAK779 and improve the understanding of various structural changes induced by the ligand in the CCR2 protein. Here, one 20 ns MD simulation was carried out on the uncomplexed CCR2 model in lipid bilayer to explore the effects of lipid membrane on the protein. Another 20 ns MD simulation was performed under the similar conditions on the docked CCR2-TAK779 complex. An alteration in the position and orientation of the ligand in binding site was observed after the simulation. Examination of protein-ligand complex suggested that TAK779 produced a greater structural change on the TM-III, TM-IV, TM-V and TM-VI than TM-I, TM-II and TM-VII. Interaction networks involving the conserved residues of uncomplexed and ‘antagonist-complexed’ CCR2 models were also examined. The major difference was observed to be the role of conserved residues of the DRY motif of TM-III and the NPxxY motif of TM-VII of CCR2.

  5. The TORC1/2 inhibitor TAK228 sensitizes atypical teratoid rhabdoid tumors to cisplatin-induced cytotoxicity.

    Science.gov (United States)

    Rubens, Jeffrey A; Wang, Sabrina Z; Price, Antoinette; Weingart, Melanie F; Allen, Sariah J; Orr, Brent A; Eberhart, Charles G; Raabe, Eric H

    2017-10-01

    Atypical teratoid/rhabdoid tumors (AT/RTs) are deadly pediatric brain tumors driven by LIN28. Mammalian target of rapamycin (mTOR) is activated in many deadly, drug-resistant cancers and governs important cellular functions such as metabolism and survival. LIN28 regulates mTOR in normal cells. We therefore hypothesized that mTOR is activated downstream of LIN28 in AT/RT, and the brain-penetrating mTOR complex 1 and 2 (mTORC1/2) kinase inhibitor TAK228 would reduce AT/RT tumorigenicity. Activation of mTOR in AT/RT was determined by measuring pS6 and pAKT (Ser473) by immunohistochemistry on tissue microarray of 18 primary AT/RT tumors. In vitro growth assays (BrdU and MTS), death assays (CC3, c-PARP by western blot), and survival curves of AT/RT orthotopic xenograft models were used to measure the efficacy of TAK228 alone and in combination with cisplatin. Lentiviral short hairpin RNA-mediated knockdown of LIN28A led to decreased mTOR activation. Primary human AT/RT had high levels of pS6 and pAKT (Ser473) in 21% and 87% of tumors by immunohistochemistry. TAK228 slowed cell growth, induced apoptosis in vitro, and nearly doubled median survival of orthotopic xenograft models of AT/RT. TAK228 combined with cisplatin synergistically slowed cell growth and enhanced cisplatin-induced apoptosis. Suppression of AKT sensitized cells to cisplatin-induced apoptosis and forced activation of AKT protected cells. Combined treatment with TAK228 and cisplatin significantly extended survival of orthotopic xenograft models of AT/RT compared with each drug alone. TAK228 has efficacy in AT/RT as a single agent and synergizes with conventional chemotherapies by sensitizing tumors to cisplatin-induced apoptosis. These results suggest TAK228 may be an effective new treatment for AT/RT. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

  6. Yeni Ürün Geliştirme Takımlarında Güven ve Takım Öğrenmesi

    Directory of Open Access Journals (Sweden)

    Volkan POLAT

    2018-01-01

    Full Text Available Farklı örgütsel seviyelerde güven ve öğrenme üzerine yapılan çalışmaların yanı sıra, güvenin takım bağlamında bilgi paylaşımı ve edinimi için önemli bir faktör olduğunu bildiren birtakım çalışmalar bulunmaktadır. Ancak bugüne kadar yeni ürün geliştirme takımlarında (new product development – NPD Takım Güveni ve Takım Öğrenmesi ilişkileri hakkında çok az tartışma yapılmıştır. Bu çalışma, Takım Güveninin, çeşitli takım faktörleri (Takım Üyesi Deneyimi, Takım İstikrarı ve Takım Otonomisi ve Takım Öğrenimi arasında arabuluculuk etkisi olup olmadığını incelemeyi amaçlamaktadır.

  7. Correlation of beam loss to residual activation in the AGS

    International Nuclear Information System (INIS)

    Brown, K.A.

    1991-01-01

    Studies of beam loss and activation at the AGS have provided a better understanding of measurements of beam loss and how they may be used to predict activation. Studies have been done in which first order correlations have been made between measured beam losses on the distributed ionization chamber system in the AGS and the health physics recorded residual activation. These studies have provided important insight into the ionization chamber system, its limitations, and its usefulness in the prediction of activation based on monitored beam loss. In recent years the AGS has run high intensity protons primarily for rare kaon decay experiments. In this mode of running the AGS typically accelerates beam from an injection momentum of 0.644 GeV/c up to a slow extracted beam (SEB) momentum of 24.2 GeV/c. The beam intensities are on the order of 4.5 x 10 13 protons per AGS cycle at injection to as high as 1.9 x 10 13 protons per AGS cycle at extraction. Residual activation varies around the AGS ring from the order of 5 mR/hour to levels of the order at 5 R/hour. The highest levels occur around the AGS beam catcher and the extraction equipment

  8. Correlation of beam loss to residual activation in the AGS

    International Nuclear Information System (INIS)

    Brown, K.A.

    1991-01-01

    Studies of beam loss and activation at the AGS have provided a better understanding of measurements of beam loss and how they may be used to predict activation. Studies have been done in which first order correlations have been made between measured beam losses on the distributed ionization chamber system in the AGS and the health physics recorded residual activation. These studies have provided important insight into the ionization chamber system, its limitations, and its usefulness in the prediction of activation based on monitored beam loss. In recent years the AGS has run high intensity protons primarily for rare known decay experiments. In this mode of running the AGS typically accelerates beam from an injection momentum of 0.644 GeV/c up to a slow extracted beam (SEB) momentum of 24.2 GeV/c. The beam intensities are on the order of 4.5 x 10 13 protons per AGS cycle at injection to as high as 1.9 x 10 13 protons per AGS cycle at extraction. Residual activation varies around the AGS ring from the order of 5 mR/hour to levels of the order at 5 R/hour. The highest levels occur around the AGS beam catcher and the extraction equipment. 7 refs., 3 figs., 2 tabs

  9. European standardization activities on residual stress analysis by neutron diffraction

    International Nuclear Information System (INIS)

    Youtsos, A.G.; Ohms, C.

    2002-01-01

    A main objective of a recently completed European research project, RESTAND - residual stress standard using neutron diffraction, was to develop industrial confidence in the application of the neutron-diffraction technique for residual stress measurement and its principal deliverable was a relevant draft code of practice. In fact this draft standard was jointly developed within RESTAND and VAMAS TWA 20 - an international pre-normative research activity. As no such standard is yet available, on the basis of this draft standard document the European Standards Committee on Non-Destructive Testing (CEN TC/138) has established a new ad hoc Work Group (AHG7). The objective of this group is the development of a European pre-standard on a 'test method for measurement of residual stress by neutron diffraction'. The document contains the proposed protocol for making the measurements. It includes the scope of the method, an outline of the technique, the calibration and measurement procedures recommended, and details of how the strain data should be analysed to calculate stresses and establish the reliability of the results obtained. (orig.)

  10. European standardization activities on residual stress analysis by neutron diffraction

    Science.gov (United States)

    Youtsos, A. G.; Ohms, C.

    A main objective of a recently completed European research project, RESTAND - residual stress standard using neutron diffraction, was to develop industrial confidence in the application of the neutron-diffraction technique for residual stress measurement and its principal deliverable was a relevant draft code of practice. In fact this draft standard was jointly developed within RESTAND and VAMAS TWA 20 - an international pre-normative research activity. As no such standard is yet available, on the basis of this draft standard document the European Standards Committee on Non-Destructive Testing (CEN TC/138) has established a new ad hoc Work Group (AHG7). The objective of this group is the development of a European pre-standard on a `test method for measurement of residual stress by neutron diffraction'. The document contains the proposed protocol for making the measurements. It includes the scope of the method, an outline of the technique, the calibration and measurement procedures recommended, and details of how the strain data should be analysed to calculate stresses and establish the reliability of the results obtained.

  11. Activated carbon from flash pyrolysis of eucalyptus residue

    Directory of Open Access Journals (Sweden)

    Grima-Olmedo C

    2016-09-01

    Full Text Available Forestry waste (eucalyptus sp was converted into activated carbon by initial flash pyrolysis followed carbonization and CO2 activation. These residues were obtained from a pilot plant in Spain that produces biofuel, the biochar represented 10–15% in weight. It was observed that the highest activation was achieved at a temperature of 800 °C, the specific surface increased with time but, on the contrary, high loss of matter was observed. At 600 °C, although there was an important increase of the specific surface and the volume of micropores, at this temperature it was observed that the activation time was not an influential parameter. Finally, at 400 °C it was observed that the activation process was not very significant. Assessing the average pore diameter it was found that the lowest value corresponded to the activation temperature of 600 °C, which indicated the development of microporosity. When the activation temperature increases up to 800 °C the pore diameter increased developing mesoporosity.

  12. Meteorological monitoring system of TÜBİTAK National Observatory

    Science.gov (United States)

    Koçak, M.; Selam, S. O.; Keskn, V.

    2004-10-01

    A custom meteorological monitoring system was constructed to reliably monitor the meteorological parameters of the site of TÜBİTAK National Observatory (TÜBİTAK: The Scientific and Technical Research Council of Turkey). The site is located on a mountain top known as Bakırlıtepe about 50 km west of the Antalya City at a height of 2547m. The system has software (C-based data acquisition/archiving structure and PHP based WEB monitoring support) and micro-controller based control electronics, fiber based custom designed encoder sensors (for wind speed and direction) and transmission lines using fiberoptic to RS232 transcievers. The constructed system can be used in any robotic telescope project for data monitoring and alert system creation.

  13. Prediction of residual metabolic activity after treatment in NSCLC patients

    International Nuclear Information System (INIS)

    Rios Velazquez, Emmanuel; Aerts, Hugo J.W.L.; Oberije, Cary; Ruysscher, Dirk De; Lambin, Philippe

    2010-01-01

    Purpose. Metabolic response assessment is often used as a surrogate of local failure and survival. Early identification of patients with residual metabolic activity is essential as this enables selection of patients who could potentially benefit from additional therapy. We report on the development of a pre-treatment prediction model for metabolic response using patient, tumor and treatment factors. Methods. One hundred and one patients with inoperable NSCLC (stage I-IV), treated with 3D conformal radical (chemo)-radiotherapy were retrospectively included in this study. All patients received a pre and post-radiotherapy fluorodeoxyglucose positron emission tomography-computed tomography FDG-PET-CT scan. The electronic medical record system and the medical patient charts were reviewed to obtain demographic, clinical, tumor and treatment data. Primary outcome measure was examined using a metabolic response assessment on a post-radiotherapy FDG-PET-CT scan. Radiotherapy was delivered in fractions of 1.8 Gy, twice a day, with a median prescribed dose of 60 Gy. Results. Overall survival was worse in patients with residual metabolic active areas compared with the patients with a complete metabolic response (p=0.0001). In univariate analysis, three variables were significantly associated with residual disease: larger primary gross tumor volume (GTVprimary, p=0.002), higher pre-treatment maximum standardized uptake value (SUV max , p=0.0005) in the primary tumor and shorter overall treatment time (OTT, p=0.046). A multivariate model including GTVprimary, SUV max , equivalent radiation dose at 2 Gy corrected for time (EQD2, T) and OTT yielded an area under the curve assessed by the leave-one-out cross validation of 0.71 (95% CI, 0.65-0.76). Conclusion. Our results confirmed the validity of metabolic response assessment as a surrogate of survival. We developed a multivariate model that is able to identify patients at risk of residual disease. These patients may benefit from

  14. TRADD, TRAF2, RIP1 and TAK1 are required for TNF-α-induced pro-labour mediators in human primary myometrial cells.

    Science.gov (United States)

    Lim, Ratana; Barker, Gillian; Lappas, Martha

    2017-07-01

    TNF-α plays a central role in the processes of human labour and delivery. This study sought to determine the role of the adaptor proteins TNFR1-associated death domain protein (TRADD), TNF receptor-associated factor 2 (TRAF2), receptor interacting protein 1 (RIP1) and transforming growth factor beta-activated kinase 1 (TAK1) in TNF-α-induced formation of pro-labour mediators. Human primary myometrial cells were transfected with siRNA against TRADD (siTRADD), TRAF2 (siTRAF2), RIP1 (siRIP1) or TAK1 (siTAK1), treated with TNF-α, and assayed for pro-inflammatory mediators expression. siTRADD, siTRAF2, siRIP1 and siTAK1 significantly decreased TNF-α-induced IL-1α, IL-1β, IL-6, IL-8, MCP-1 mRNA expression and release of IL-6, IL-8 and MCP-1; and cyclooxygenase (COX)-2 expression and release of prostaglandin PGF 2α . There was a significant attenuation of TNF-α-induced expression of adhesion molecules ICAM-1 and VCAM-1 mRNA with siTRADD, siTRAF2 or siRIP1. siTRADD and siRIP1 significantly attenuated TNF-α-induced MMP-9 mRNA expression and release and nuclear factor κB (NF-κB) transcriptional activity. There was a significant increase in TNF-α-induced sVCAM-1 release, MMP-9 mRNA expression and NF-κB activity with siTAK1. TRADD, TRAF2, RIP1 and TAK1 are involved in TNF-α signalling in human myometrium. Further studies are required to determine whether inhibition of these proteins can prevent preterm birth. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  15. An intention-activation account of residual switch costs

    NARCIS (Netherlands)

    De Jong, R; Monsell, S; Driver, J

    2000-01-01

    Residual switch costs are performance costs associated with a shift of task that persist even when there is ample time to prepare in advance for the new task. I present a mixture-model approach for evaluating the contributions of two possible causes of residual switch costs: (1) failures to take

  16. Momordica charantia Inhibits Inflammatory Responses in Murine Macrophages via Suppression of TAK1.

    Science.gov (United States)

    Yang, Woo Seok; Yang, Eunju; Kim, Min-Jeong; Jeong, Deok; Yoon, Deok Hyo; Sung, Gi-Ho; Lee, Seungihm; Yoo, Byong Chul; Yeo, Seung-Gu; Cho, Jae Youl

    2018-01-01

    Momordica charantia known as bitter melon is a representative medicinal plant reported to exhibit numerous pharmacological activities such as antibacterial, antidiabetic, anti-inflammatory, anti-oxidant, antitumor, and hypoglycemic actions. Although this plant has high ethnopharmacological value for treating inflammatory diseases, the molecular mechanisms by which it inhibits the inflammatory response are not fully understood. In this study, we aim to identify the anti-inflammatory mechanism of this plant. To this end, we studied the effects of its methanol extract (Mc-ME) on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Specifically, we evaluated nitric oxide (NO) production, mRNA expression of inflammatory genes, luciferase reporter gene activity, and putative molecular targets. Mc-ME blocked NO production in a dose-dependent manner in RAW264.7 cells; importantly, no cytotoxicity was observed. Moreover, the mRNA expression levels of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 were decreased by Mc-ME treatment in a dose-dependent manner. Luciferase assays and nuclear lysate immunoblotting analyses strongly indicated that Mc-ME decreases the levels of p65 [a nuclear factor (NF)-[Formula: see text]B subunit] and c-Fos [an activator protein (AP)-1 subunit]. Whole lysate immunoblotting assays, luciferase assays, and overexpression experiments suggested that transforming growth factor [Formula: see text]-activated kinase 1 (TAK1) is targeted by Mc-ME, thereby suppressing NF-[Formula: see text]B and AP-1 activity via downregulation of extracellular signal-regulated kinases (ERKs) and AKT. These results strongly suggest that Mc-ME exerts its anti-inflammatory activity by reducing the action of TAK1, which also affects the activation of NF-[Formula: see text]B and AP-1.

  17. Leaching and residual activity of imidazolinone herbicides in lowland soils

    Directory of Open Access Journals (Sweden)

    João Paulo Refatti

    Full Text Available ABSTRACT: Herbicides used in the Clearfield® rice (Oryza sativa L. production system have a potential for leaching. This can result in contamination of underground water resources and cause injury to not tolerant crops that are sown in a succession and/or crop rotation. The objective of this study was to determine the leaching potential and the residual activity of the herbicides used in the Clearfield® rice system. The experiment was conducted over a period of two years and consisted of conducting a field test to be followed by two bioassays with a year of difference between their implementation. Initially an experiment was conducted in lowland area where it was planted the cultivar of rice ‘PUITA INTA CL’. Approximately one and two years thereafter, soil samples from each plot were collected at intervals of 5cm to a depth of 30cm (B factor for the bioassay to evaluate persistence of herbicides. Factor A was composed of mixtures formulated of imazethapyr + imazapic (75 + 25g a.i. L-1, imazapyr + imazapic (525 + 175g a.i. kg-1 in two doses, imazethapyr (100g a.i. L-1 and treatment control without application. Basing on results, it was concluded that the mixtures imazethapyr + imazapic, imazapyr + imazapic and imazethapyr leached into the soil, reaching depths of up to 25cm in lowland soil. Imidazolinone herbicides used today in the irrigated rice Clearfield® system are persistent in soil, and their phytotoxic activity can be observed up to two years after application.

  18. Effect of a glycine residue insertion into crustacean hyperglycemic hormone on hormonal activity

    OpenAIRE

    Katayama, Hidekazu; Nagasawa, Hiromichi

    2004-01-01

    Crustacean hyperglycemic hormone (CHH) and molt-inhibiting hormone (MIH) have similar amino acid sequences and therefore comprise a peptide family referred to as the CHH family. All MIHs unexceptionally have an additional glycine residue at position 12, which is lacking in all CHHs. In order to understand the relevance of the absence of the glycine residue for hyperglycemic activity, a mutant CHH having a glycine residue insertion was prepared, and its hyperglycemic activity was assessed. Thi...

  19. Effect of a Glycine Residue Insertion into Crustacean Hyperglycemic Hormone on Hormonal Activity(Endocrinology)

    OpenAIRE

    Hidekazu, Katayama; Hiromichi, Nagasawa; Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo; Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo

    2004-01-01

    Crustacean hyperglycemic hormone (CHH) and molt-inhibiting hormone (MIH) have similar amino acid sequences and therefore comprise a peptide family referred to as the CHH family. All MIHs unexceptionally have an additional glycine residue at position 12, which is lacking in all CHHs. In order to understand the relevance of the absence of the glycine residue for hyperglycemic activity, a mutant CHH having a glycine residue insertion was prepared, and its hyperglycemic activity was assessed. Thi...

  20. Drug evaluation: TAK-475--an oral inhibitor of squalene synthase for hyperlipidemia.

    Science.gov (United States)

    Burnett, John R

    2006-09-01

    Takeda Pharmaceutical Co Ltd is developing TAK-475, a squalene synthetase inhibitor from a series of 4,1-benzoxazepine-3-acetic acid derivatives, for the potential oral treatment of hyperlipidemia. By March 2005, TAK-475 was undergoing phase III clinical trials in the US and Europe.

  1. Effect of soil-bound residues of malathion on microbial activities

    International Nuclear Information System (INIS)

    Hussain, A.; Iqbal, Z.; Asi, M.R.; Tahira, R.; Chudhary, J.A.

    2001-01-01

    The effect of soil-bound residues of malathion on CO/sub 2/ evolution, dehydrogenase activity and some nitrogen transformations in a loam soil was investigated under laboratory conditions. The soil samples containing bound residues arising from 10 mg g-1 of the applied malathion were mixed in equal quantity with fresh soil and compared with solvent extracted control soil without bound residues (extracted in the same way as soil containing bound residues). Another control comprising un extracted fresh soil without bound residues was also kept to study the effect of solvent extraction on the biological activity. Rate of Carbon mineralization (CO/sub 2/ evolution) was decreased in the presence of soil-bound residues of malathion. Bound residues also affected dehydrogenase activity of soil. Over 40% inhibition of dehydrogenase activity was observed after 4 days and the inhibition persisted at least for 12 days. Nitrogen mineralization was stimulated in soil containing bound residues of malathion and this stimulatory effect increased with time of incubation. Nitrification was partially inhibited in the presence of soil-bound residues of malathion. The inhibitory effect of the soil-bound residues on nitrification did not show much variation with time. The soil-bound residues did not affect denitrification rate (N/sub 2/O evolution). Nitrogen fixation (acetylene reduction) was partially inhibited in soil amended with bound residues of malathion and the inhibitory effect persisted for at least one week. In general, soil bound residues of malathion inhibited CO/sub 2/ evolution, dehydrogenase activity, nitrification and nitrogen fixation while mineralization of nitrogen was stimulated. Denitrification was not affected by the applied insecticide. (author)

  2. Enhancement of transglutaminase activity by NMR identification of its flexible residues affecting the active site.

    Science.gov (United States)

    Shimba, Nobuhisa; Shinohara, Mina; Yokoyama, Kei ichi; Kashiwagi, Tatsuki; Ishikawa, Kohki; Ejima, Daisuke; Suzuki, Ei ichiro

    2002-04-24

    Incorporation of inter- or intramolecular covalent cross-links into food proteins with microbial transglutaminase (MTG) improves the physical and textural properties of many food proteins, such as tofu, boiled fish paste, and sausage. By using nuclear magnetic resonance, we have shown that the residues exhibiting relatively high flexibility in MTG are localized in the N-terminal region; however, the N-terminal region influences the microenvironment of the active site. These results suggest that the N-terminal region is not of primary importance for the global fold, but influences the substrate binding. Therefore, in order to increase the transglutaminase activity, the N-terminal residues were chosen as candidates for site-directed replacement and deletion. We obtained several mutants with higher activity, del1-2, del1-3, and S2R. We propose a strategy for enzyme engineering targeted toward flexible regions involved in the enzymatic activity. In addition, we also briefly describe how the number of glutamine residues in a substrate protein can be increased by mixing more than two kinds of TGases with different substrate specificities.

  3. Active-site residues of procarboxypeptidase Y are accessible to chemical modification

    DEFF Research Database (Denmark)

    Sørensen, S O; Winther, Jakob R.

    1994-01-01

    The accessibility of the active-site cleft of procarboxypeptidase Y from Saccharomyces cerevisiae has been studied by chemical modifications of two specific amino-acid residues. Previous studies have shown that these residues, Cys-341 and Met-398 in the mature enzyme, are located in the S1 and S'1...

  4. A study on the neoasozine residues in rice grain by neutron activation method

    International Nuclear Information System (INIS)

    Kim, Y.H.; Lee, K.J.; Lee, S.R.

    1981-01-01

    Residues of neoasozine in rice grain were determined by neutron activation and colorimetric techniques. Twice application of the chemical before flowering did not lead to any increased residue level while 4-times application resulted in significant increase in the residue level up to 0.54 - 0.75 mg As 2 O 3 /kg. The partition ratio of arsenic residues into polished rice grain and bran was 73 : 27 in 100 % polishing while most of the residues in the bran was transferred to oil cake fraction during solvent extraction, reaching up to 2.9 mg As 2 O 3 /kg. The neutron activation technique was advantageous because of its high sensitivity and the smaller sample amounts required for analysis. (author)

  5. A method for the determination of residual beta activity in drinking water samples

    Energy Technology Data Exchange (ETDEWEB)

    Idoeta, R. [Dpto. Ingenieria Nuclear y Mecanica de Fluidos, E. T. S. Ingenieria de Bilbao - Universidad del Pais Vasco (UPV/EHU), Alda. Urquijo s/n. 48013 Bilbao (Spain)], E-mail: raquel.idoeta@ehu.es; Herranz, M.; Abelairas, A.; Legarda, F. [Dpto. Ingenieria Nuclear y Mecanica de Fluidos, E. T. S. Ingenieria de Bilbao - Universidad del Pais Vasco (UPV/EHU), Alda. Urquijo s/n. 48013 Bilbao (Spain)

    2007-09-15

    The determination of residual beta activity in drinking water is usually needed in most monitoring programs. In this work a procedure for its determination is described and expressions for the calculations of detection limits and uncertainties are proposed.

  6. A method for the determination of residual beta activity in drinking water samples

    International Nuclear Information System (INIS)

    Idoeta, R.; Herranz, M.; Abelairas, A.; Legarda, F.

    2007-01-01

    The determination of residual beta activity in drinking water is usually needed in most monitoring programs. In this work a procedure for its determination is described and expressions for the calculations of detection limits and uncertainties are proposed

  7. Residual activities of 99mTc-labelled radiopharmaceuticals in routine nuclear medicine practice.

    Science.gov (United States)

    Stavrou, Petros Z; Papachristou, Maria; Persakis, Evangelos; Kouvelis, Konstantinos; Datseris, Ioannis E

    2016-06-01

    In this study, we present a series of common Tc-labelled radiopharmaceuticals administrations performed in a busy nuclear medicine department within a 1-year timeframe in routine practice. The main objective is to identify the true activity administered to the patient and whether that is in compliance with the EANM guidelines, where applicable, and/or with the department's protocols. As a secondary objective, we aimed to assess whether the volume of the injected dose correlates to the percentage of residual activity measured after injection for each studied radiotracer. A number of commonly used Tc-labelled radiopharmaceuticals including Tc-pertechnetate, Tc-methylenediphosphonic acid, Tc-hydroxymethylenediphosphonic acid, Tc-3,3-diphosphono-1,2-propanedicarboxylic acid, Tc-dimercaptosuccinic acid (DMSA), Tc-mercaptoacetyltriglycine, Tc-2-methoxyisobutylisonitrile (sestamibi), Tc-1,2-bis[bis(2-ethoxyethyl)phosphino]ethane (tetrofosmin) and Tc-HYNIC-Tyr-octreotide (Tektrotyd) were assessed. All data were collected prospectively within a 1-year period under routine medical practice and included (a) injected activity, (b) residual activity, (c) percentage of residual activity (residual activity/injected activity×100), (d) effective activity (injected activity minus the residual activity), (e) injected volume, (f) time of injection and (g) type of procedure. In the 1837 collected measurements, the average percentage of residual activity was 13% (95% confidence interval: 12.75-13.39%). The mean effective activities were within the recommended range by the EANM guidelines, where applicable, or did not exceed the recommended doses from the department's protocols. In cases where actual injected activity was lower than the minimum suggested by the department's protocols, the imaging quality was not compromised. There was a negative correlation and thus an inverse relationship between the percentage of residual activity and administered volume for Tc-pertechnetate, Teceos

  8. The performance of activated carbons from sugarcane bagasse, babassu, and coconut shells in removing residual chlorine

    Directory of Open Access Journals (Sweden)

    E. F. Jaguaribe

    2005-03-01

    Full Text Available The capacity of activated carbons obtained from different raw materials, such as sugarcane bagasse, babassu (Orbygnia speciosa, and coconut (Cocus nucifera shells, to remove residual chlorine is studied. The influence of particle size and time of contact between particles of activated carbon and the chlorinated solution were taken into account. The adsorptive properties of the activated carbons were measured by gas adsorption (BET method, using an ASAP 2010 porosimeter, and liquid phase adsorption, employing iodine and methylene blue adsorbates. The activated carbon from sugarcane bagasse was the only adsorbent capable of removing 100% of the residual chlorine.

  9. Identification of key neoculin residues responsible for the binding and activation of the sweet taste receptor

    Science.gov (United States)

    Koizumi, Taichi; Terada, Tohru; Nakajima, Ken-ichiro; Kojima, Masaki; Koshiba, Seizo; Matsumura, Yoshitaka; Kaneda, Kohei; Asakura, Tomiko; Shimizu-Ibuka, Akiko; Abe, Keiko; Misaka, Takumi

    2015-01-01

    Neoculin (NCL) is a heterodimeric protein isolated from the edible fruit of Curculigo latifolia. It exerts a taste-modifying activity by converting sourness to sweetness. We previously demonstrated that NCL changes its action on the human sweet receptor hT1R2-hT1R3 from antagonism to agonism as the pH changes from neutral to acidic values, and that the histidine residues of NCL molecule play critical roles in this pH-dependent functional change. Here, we comprehensively screened key amino acid residues of NCL using nuclear magnetic resonance (NMR) spectroscopy and alanine scanning mutagenesis. We found that the mutations of Arg48, Tyr65, Val72 and Phe94 of NCL basic subunit increased or decreased both the antagonist and agonist activities. The mutations had only a slight effect on the pH-dependent functional change. These residues should determine the affinity of NCL for the receptor regardless of pH. Their locations were separated from the histidine residues responsible for the pH-dependent functional change in the tertiary structure. From these results, we concluded that NCL interacts with hT1R2-hT1R3 through a pH-independent affinity interface including the four residues and a pH-dependent activation interface including the histidine residues. Thus, the receptor activation is induced by local structural changes in the pH-dependent interface. PMID:26263392

  10. The balance of flexibility and rigidity in the active site residues of hen egg white lysozyme

    International Nuclear Information System (INIS)

    Qi Jian-Xun; Jiang Fan

    2011-01-01

    The crystallographic temperature factors (B factor) of individual atoms contain important information about the thermal motion of the atoms in a macromolecule. Previously the theory of flexibility of active site has been established based on the observation that the enzyme activity is sensitive to low concentration denaturing agents. It has been found that the loss of enzyme activity occurs well before the disruption of the three-dimensional structural scaffold of the enzyme. To test the theory of conformational flexibility of enzyme active site, crystal structures were perturbed by soaking in low concentration guanidine hydrochloride solutions. It was found that many lysozyme crystals tested could still diffract until the concentration of guanidine hydrochloride reached 3 M. It was also found that the B factors averaged over individually collected data sets were more accurate. Thus it suggested that accurate measurement of crystal temperature factors could be achieved for medium-high or even medium resolution crystals by averaging over multiple data sets. Furthermore, we found that the correctly predicted active sites included not only the more flexible residues, but also some more rigid residues. Both the flexible and the rigid residues in the active site played an important role in forming the active site residue network, covering the majority of the substrate binding residues. Therefore, this experimental prediction method may be useful for characterizing the binding site and the function of a protein, such as drug targeting. (interdisciplinary physics and related areas of science and technology)

  11. Dissection of Functional Residues in Receptor Activity-Modifying Proteins Through Phylogenetic and Statistical Analyses

    Directory of Open Access Journals (Sweden)

    Alfonso Benítez-Páez

    2008-01-01

    Full Text Available Type I and type-II functional divergences have been stated to highlight specific residues carrying out differential functions in evolutionary-divergent protein clusters from a single common ancestor. Briefly, type I analysis is based on residue constraints reflecting a gain of function just in one cluster of an entire family of proteins; while the type-II approach is based on residue constraints showing a different chemical nature in every cluster of a protein family. This last evidence is understood as differential functionality among clusters. The Receptor Activity-Modifying Proteins constitute a family characterized by its paralogous distribution in vertebrates. They are known as G-Protein Coupled Receptor modulators. Although several studies have determined their involvement in ligand binding, specificity, and enhancement of signal transduction, the responsible residues supporting those functions are unclear. Using different bioinformatic approaches, we predicted residues involved in different RAMP functional tasks. Many residues localized in an extracellular coil of RAMP proteins were predicted to be under functional divergence suggesting a gain of function in their respective proteins. Interestingly, the transmembrane region also showed important results for residues playing relevant roles where most of them showed a biased distribution on the structure. A relevant role was conferred by the enrichment of type-II residues observed in their sequences. We show a collection of residues explaining possible gain of function and differential functionality in RAMP proteins. These residues are still experimentally unexplored with regards to functionality. Finally, an evolutionary history could be discerned. Mainly, the RAMP2 cluster has evolved in a higher manner than other RAMP clusters. However, a deacceleration in the aminoacid substitution rate of RAMP2 was observed in mammals. Such effect could be caused by the co-evolution of ligands and

  12. Residual deposits (residual soil)

    International Nuclear Information System (INIS)

    Khasanov, A.Kh.

    1988-01-01

    Residual soil deposits is accumulation of new formate ore minerals on the earth surface, arise as a result of chemical decomposition of rocks. As is well known, at the hyper genes zone under the influence of different factors (water, carbonic acid, organic acids, oxygen, microorganism activity) passes chemical weathering of rocks. Residual soil deposits forming depends from complex of geologic and climatic factors and also from composition and physical and chemical properties of initial rocks

  13. The effects of residual platelets in plasma on plasminogen activator inhibitor-1 and plasminogen activator inhibitor-1-related assays

    NARCIS (Netherlands)

    M. Pieters (Marlien); S.A. Barnard (Sunelle A.); D.T. Loots (Du Toit); D.C. Rijken (Dingeman)

    2017-01-01

    textabstractDue to controversial evidence in the literature pertaining to the activity of plasminogen activator inhibitor-1 in platelets, we examined the effects of residual platelets present in plasma (a potential pre-analytical variable) on various plasminogen activator inhibitor-1 and plasminogen

  14. Controlling the activity of the Tec kinase Itk by mutation of the phenylalanine gatekeeper residue.

    Science.gov (United States)

    Joseph, Raji E; Andreotti, Amy H

    2011-01-18

    The regulatory spine is a set of conserved residues that are assembled and disassembled upon activation and inactivation of kinases. We recently identified the regulatory spine within the immunologically important Tec family kinases and have shown that in addition to the core spine residues within the kinase domain itself, contributions from the SH2-kinase linker region result in an extended spine structure for this kinase family. Disruption of the regulatory spine, either by mutation or by removal of the amino-terminal SH2-kinase linker region or by mutation of core spine residues, leads to inactivation of the Tec kinases. With a focus on the Tec family members, Itk and Btk, we now show that the gatekeeper residue is also critical for the assembly of the regulatory spine. Mutation of the bulky Itk F434 gatekeeper residue to alanine or glycine inactivates Itk. The activity of the Itk F434A mutant can be recovered by a secondary site mutation within the N-terminal lobe, specifically L432I. The Itk L432I mutation likely rescues the activity of the gatekeeper F434A mutation by promoting the assembly of the regulatory spine. We also show that mutation of the Itk and Btk gatekeeper residues to methionine is sufficient to activate the isolated kinase domains of Tec kinases in the absence of the amino-terminal SH2-kinase linker. Thus, shifting the conformational equilibrium between the assembled and disassembled states of the regulatory spine by changing the nature of the gatekeeper residue is key to regulating the activity of Tec kinases.

  15. Radiological characterization and evaluation of high volume bauxite residue alkali activated concretes.

    Science.gov (United States)

    Croymans, Tom; Schroeyers, Wouter; Krivenko, Pavel; Kovalchuk, Oleksandr; Pasko, Anton; Hult, Mikael; Marissens, Gerd; Lutter, Guillaume; Schreurs, Sonja

    2017-03-01

    Bauxite residue, also known as red mud, can be used as an aggregate in concrete products. The study involves the radiological characterization of different types of concretes containing bauxite residue from Ukraine. The activity concentrations of radionuclides from the 238 U, 232 Th decay series and 40 K were determined for concrete mixture samples incorporating 30, 40, 50, 60, 75, 85 and 90% (by mass) of bauxite residue using gamma-ray spectrometry with a HPGe detector. The studied bauxite residue can, from a radiological point of view using activity concentration indexes developed by Markkanen, be used in concrete for building materials and in road construction, even in percentages reaching 90% (by mass). However, when also occupational exposure is considered it is recommended to incorporate less than 75% (by mass) of Ukrainian bauxite residue during the construction of buildings in order to keep the dose to workers below the dose criterion used by Radiation Protection (RP) 122 (0.3 mSv/a). Considering RP122 for evaluation of the total effective dose to workers no restrictions are required for the use of the Ukrainian bauxite residue in road construction. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Simulation analysis of ASM/Takács models in the BSM1 configuration.

    Science.gov (United States)

    Sorour, M T; Bahgat, L M F

    2006-10-01

    The present paper summarizes the outlines of a simulation analysis study conducted on the BSM1 (formerly the COST benchmark) configuration using activated sludge models (ASM1, ASM2d and ASM3) coupled to Takács settler model. The prime objective was to develop reliable simulation software programs to implement these complex models according to the working conditions of a realistic plant. The analysis focused on comparing the steady state predictions of models ASM1/ ASM3 when imposed to pulse/ step type disturbances on wastewater characteristics and control variables, then on assessing the capability of the simulated configuration for bio-P removal using model ASM2d. Section 1 of the paper briefly presents problem definition/ solution approach while section 2 demonstrates some examples showing the main indications of the simulation analysis. ASM1/ ASM3 predictions indicate the presence of some significant differences between both models that could be related to their underlying concepts. ASM2d simulations show that adverse effects on the permissible limit of effluent's ammonia concentration should be expected when the plant is operated to achieve dual nutrient removal.

  17. Identification of the amino acid residues essential for proteolytic activity in an archaeal signal peptide peptidase.

    Science.gov (United States)

    Matsumi, Rie; Atomi, Haruyuki; Imanaka, Tadayuki

    2006-04-14

    Signal peptide peptidases (SPPs) are enzymes involved in the initial degradation of signal peptides after they are released from the precursor proteins by signal peptidases. In contrast to the eukaryotic enzymes that are aspartate peptidases, the catalytic mechanisms of prokaryotic SPPs had not been known. In this study on the SPP from the hyperthermophilic archaeon Thermococcus kodakaraensis (SppA(Tk)), we have identified amino acid residues that are essential for the peptidase activity of the enzyme. DeltaN54SppA(Tk), a truncated protein without the N-terminal 54 residues and putative transmembrane domain, exhibits high peptidase activity, and was used as the wild-type protein. Sixteen residues, highly conserved among archaeal SPP homologue sequences, were selected and replaced by alanine residues. The mutations S162A and K214A were found to abolish peptidase activity of the protein, whereas all other mutant proteins displayed activity to various extents. The results indicated the function of Ser(162) as the nucleophilic serine and that of Lys(214) as the general base, comprising a Ser/Lys catalytic dyad in SppA(Tk). Kinetic analyses indicated that Ser(184), His(191) Lys(209), Asp(215), and Arg(221) supported peptidase activity. Intriguingly, a large number of mutations led to an increase in activity levels of the enzyme. In particular, mutations in Ser(128) and Tyr(165) not only increased activity levels but also broadened the substrate specificity of SppA(Tk), suggesting that these residues may be present to prevent the enzyme from cleaving unintended peptide/protein substrates in the cell. A detailed alignment of prokaryotic SPP sequences strongly suggested that the majority of archaeal enzymes, along with the bacterial enzyme from Bacillus subtilis, adopt the same catalytic mechanism for peptide hydrolysis.

  18. Evidence of the Participation of Remote Residues in the Catalytic Activity of Co-Type Nitrile Hydratase from Pseudomonas putida

    Energy Technology Data Exchange (ETDEWEB)

    Brodkin, Heather R.; Novak, Walter R.P.; Milne, Amy C.; D’Aquino, J. Alejandro; Karabacak, N.M.; Goldberg, Ilana G.; Agar, Jeffrey N.; Payne, Mark S.; Petsko, Gregory A.; Ondrechen, Mary Jo; Ringe, Dagmar (Brandeis); (E.I. DuPont); (NEU)

    2011-09-28

    Active sites may be regarded as layers of residues, whereby the residues that interact directly with substrate also interact with residues in a second shell and these in turn interact with residues in a third shell. These residues in the second and third layers may have distinct roles in maintaining the essential chemical properties of the first-shell catalytic residues, particularly their spatial arrangement relative to the substrate binding pocket, and their electrostatic and dynamic properties. The extent to which these remote residues participate in catalysis and precisely how they affect first-shell residues remains unexplored. To improve our understanding of the roles of second- and third-shell residues in catalysis, we used THEMATICS to identify residues in the second and third shells of the Co-type nitrile hydratase from Pseudomonas putida (ppNHase) that may be important for catalysis. Five of these predicted residues, and three additional, conserved residues that were not predicted, have been conservatively mutated, and their effects have been studied both kinetically and structurally. The eight residues have no direct contact with the active site metal ion or bound substrate. These results demonstrate that three of the predicted second-shell residues ({alpha}-Asp164, {beta}-Glu56, and {beta}-His147) and one predicted third-shell residue ({beta}-His71) have significant effects on the catalytic efficiency of the enzyme. One of the predicted residues ({alpha}-Glu168) and the three residues not predicted ({alpha}-Arg170, {alpha}-Tyr171, and {beta}-Tyr215) do not have any significant effects on the catalytic efficiency of the enzyme.

  19. Biotransformation of fluoroquinolone antibiotics by ligninolytic fungi - Metabolites, enzymes and residual antibacterial activity

    Czech Academy of Sciences Publication Activity Database

    Čvančarová, Monika; Moeder, M.; Filipová, Alena; Cajthaml, Tomáš

    2015-01-01

    Roč. 136, OCT 2015 (2015), s. 311-320 ISSN 0045-6535 R&D Projects: GA TA ČR TE01020218; GA ČR GA13-28283S Institutional support: RVO:61388971 Keywords : Fluoroquinolone antibiotics * White rot fungi * Residual antibacterial activity Subject RIV: EE - Microbiology, Virology Impact factor: 3.698, year: 2015

  20. Molecular Basis for Enzymatic Sulfite Oxidation -- HOW THREE CONSERVED ACTIVE SITE RESIDUES SHAPE ENZYME ACTIVITY

    Energy Technology Data Exchange (ETDEWEB)

    Bailey, Susan; Rapson, Trevor; Johnson-Winters, Kayunta; Astashkin, Andrei; Enemark, John; Kappler, Ulrike

    2008-11-10

    Sulfite dehydrogenases (SDHs) catalyze the oxidation and detoxification of sulfite to sulfate, a reaction critical to all forms of life. Sulfite-oxidizing enzymes contain three conserved active site amino acids (Arg-55, His-57, and Tyr-236) that are crucial for catalytic competency. Here we have studied the kinetic and structural effects of two novel and one previously reported substitution (R55M, H57A, Y236F) in these residues on SDH catalysis. Both Arg-55 and His-57 were found to have key roles in substrate binding. An R55M substitution increased Km(sulfite)(app) by 2-3 orders of magnitude, whereas His-57 was required for maintaining a high substrate affinity at low pH when the imidazole ring is fully protonated. This effect may be mediated by interactions of His-57 with Arg-55 that stabilize the position of the Arg-55 side chain or, alternatively, may reflect changes in the protonation state of sulfite. Unlike what is seen for SDHWT and SDHY236F, the catalytic turnover rates of SDHR55M and SDHH57A are relatively insensitive to pH (~;;60 and 200 s-1, respectively). On the structural level, striking kinetic effects appeared to correlate with disorder (in SDHH57A and SDHY236F) or absence of Arg-55 (SDHR55M), suggesting that Arg-55 and the hydrogen bonding interactions it engages in are crucial for substrate binding and catalysis. The structure of SDHR55M has sulfate bound at the active site, a fact that coincides with a significant increase in the inhibitory effect of sulfate in SDHR55M. Thus, Arg-55 also appears to be involved in enabling discrimination between the substrate and product in SDH.

  1. Determination of Activated Carbon Residual Life using a Microwave Cavity Resonator

    International Nuclear Information System (INIS)

    Mason, A; Wylie, S; Shaw, A; Al-Shamma'a, A I; Thomas, A; Keele, H

    2011-01-01

    This paper presents the continuation of work conducted jointly between Dstl and LJMU. This unique body of work has been, largely, concerned with detecting the residual life of high performance filter materials using electromagnetic (EM) waves within a resonant cavity. Past work has considered both HEPA [1] and ASZM-TEDA[2] activated carbon filter materials. This paper continues the later work, considering the response of ASZM-TEDA activated carbon through the co-ageing of two distinct batches of the material. The paper briefly introduces activated carbon, discusses theory relevant to the work and the methodology used for investigation. A comprehensive set of results is included which seek to validate this technique for determining the residual lifespan of activated carbon.

  2. Chaperone-like activity of β-casein and its effect on residual in vitro activity of horseradish peroxidase

    DEFF Research Database (Denmark)

    Sulewska, Anna Maria; Olsen, Karsten; Sørensen, Jens Christian

    2014-01-01

    , as similar experiment with bovine serum albumin resulted in residual activity of horseradish peroxidase that was significantly lower than without any addition. The effect of β-casein on HRP disappears when pH is below the isoelectric point of β-casein. It was also proven by light scattering studies that β...... proteins. Incubating HRP (0.1 mg mL-1) for 10 min at 72 °C resulted in residual activity of 59 ± 5%, while addition of 1 mg mL-1 β-casein resulted in increase in residual activity up to 85 ± 1%. Increased residual activity is not merely attributed to an effect of higher total protein concentration......-casein interacts with horseradish peroxidase when the temperature was increased from 25 to 70 °C whereas interactions seem to cease when temperature was lowered back to 25 °C. This study highlights how specific proteins can influence enzyme activity, which is of potential importance for various industries...

  3. Production of energy and activated carbon from agri-residue: sunflower seed example.

    Science.gov (United States)

    Donaldson, Adam A; Kadakia, Parag; Gupta, Murlidhar; Zhang, Zisheng

    2012-09-01

    In this work, a biomass processing facility is designed and simulated for the annual conversion of 77 ktons of sunflower residue into electricity and activated carbon. The residue is initially pyrolized to produce low hydrocarbon gases (35 wt%), bio-oils (30 wt%), and char (35 wt%). The gases and bio-oils are separated and combusted to generate high pressure steam, electricity, and steam for conversion of char into activated carbon. Assuming 35% of the char's mass is lost during activation, the proposed process produces 15.6 ktons activated carbon and 5.5 ktons ash annually, while generating 10.2 MW of electricity. Economic analysis of the proposed facility yielded capital costs of $31.64 million, annual operating costs of $31.58 million, and a yearly gross revenue of $38.9 million. A discounted payback period of 6.1 years was determined for the current design, extending to 10 years if the facility were operated at 75% capacity. While the proposed process appears to be economically viable, profitability is highly sensitive to the selling price of electricity and activated carbon, highlighting the need for additional research into the pyrolysis reactor design, char/ash separation techniques, and the quality of activated carbon obtained using char from sunflower residue pyrolysis.

  4. Antitumor effects of the investigational selective MEK inhibitor TAK733 against cutaneous and uveal melanoma cell lines

    Directory of Open Access Journals (Sweden)

    von Euw Erika

    2012-04-01

    Full Text Available Abstract Background TAK733 is a novel allosteric, non-ATP-binding, inhibitor of the BRAF substrates MEK-1/2. Methods The growth inhibitory effects of TAK733 were assessed in a panel of 27 cutaneous and five uveal melanoma cell lines genotyped for driver oncogenic mutations. Flow cytometry, Western blots and metabolic tracer uptake assays were used to characterize the changes induced by exposure to TAK733. Results Fourteen cutaneous melanoma cell lines with different driver mutations were sensitive to the antiproliferative effects of TAK733, with a higher proportion of BRAFV600E mutant cell lines being highly sensitive with IC50s below 1 nM. The five uveal melanoma cell lines had GNAQ or GNA11 mutations and were either moderately or highly sensitive to TAK733. The tested cell lines wild type for NRAS, BRAF, GNAQ and GNA11 driver mutations were moderately to highly resistant to TAK733. TAK733 led to a decrease in pERK and G1 arrest in most of these melanoma cell lines regardless of their origin, driver oncogenic mutations and in vitro sensitivity to TAK733. MEK inhibition resulted in increase in pMEK more prominently in NRASQ61L mutant and GNAQ mutant cell lines than in BRAFV600E mutant cell lines. Uptake of the metabolic tracers FDG and FLT was inhibited by TAK733 in a manner that closely paralleled the in vitro sensitivity assays. Conclusions The MEK inhibitor TAK733 has antitumor properties in melanoma cell lines with different oncogenic mutations and these effects could be detectable by differential metabolic tracer uptake.

  5. An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues

    Directory of Open Access Journals (Sweden)

    Eveline Queiroz de Pinho Tavares

    2013-01-01

    Full Text Available Aspergillus nidulans is poorly exploited as a source of enzymes for lignocellulosic residues degradation for biotechnological purposes. This work describes the A. nidulans Endoglucanase A heterologous expression in Pichia pastoris, the purification and biochemical characterization of the recombinant enzyme. Active recombinant endoglucanase A (rEG A was efficiently secreted as a 35 kDa protein which was purified through a two-step chromatography procedure. The highest enzyme activity was detected at 50°C/pH 4. rEG A retained 100% of activity when incubated at 45 and 55°C for 72 h. Purified rEG A kinetic parameters towards CMC were determined as Km=27.5±4.33 mg/mL, Vmax=1.185±0.11 mmol/min, and 55.8 IU (international units/mg specific activity. Recombinant P. pastoris supernatant presented hydrolytic activity towards lignocellulosic residues such as banana stalk, sugarcane bagasse, soybean residues, and corn straw. These data indicate that rEG A is suitable for plant biomass conversion into products of commercial importance, such as second-generation fuel ethanol.

  6. Key amino acid residues for the endo-processive activity of GH74 xyloglucanase.

    Science.gov (United States)

    Matsuzawa, Tomohiko; Saito, Yuji; Yaoi, Katsuro

    2014-05-02

    Unlike endo-dissociative-xyloglucanases, Paenibacillus XEG74 is an endo-processive xyloglucanase that contains four unique tryptophan residues in the negative subsites (W61 and W64) and the positive subsites (W318 and W319), as indicated by three-dimensional homology modelling. Selective replacement of the positive subsite residues with alanine mutations reduced the degree of processive activity and resulted in the more endo-dissociative-activity. The results showed that W318 and W319, which are found in the positive subsites, are essential for processive degradation and are responsible for maintaining binding interactions with xyloglucan polysaccharide through a stacking effect. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  7. Structural plasticity among glutathione transferase Phi members: natural combination of catalytic residues confers dual biochemical activities.

    Science.gov (United States)

    Pégeot, Henri; Mathiot, Sandrine; Perrot, Thomas; Gense, Frédéric; Hecker, Arnaud; Didierjean, Claude; Rouhier, Nicolas

    2017-08-01

    The glutathione transferase (GST) gene family is divided into 14 classes in photosynthetic organisms. Among them, the Phi class (GSTF) is composed of a large number of genes that are often induced in response to environmental constraints due to their ability to detoxify xenobiotics, to their peroxidase activity and to their involvement in the biosynthesis and/or transport of secondary metabolites. However, the exact functions of GSTFs from many plants including Populus trichocarpa are unknown. Here, following GSTF1 characterization, we have performed a comparative analysis of the seven other GSTFs found in poplar by systematically evaluating the biochemical and enzymatic properties of the corresponding recombinant proteins and of variants mutated for active site residues and by determining the three-dimensional structures of several representatives. Owing to the presence of a cysteine with a pK a value around 5 in their active site, GSTF3, F7, and F8 displayed a thiol transferase activity in addition to the usual glutathione transferase and peroxidase activities. From structural analyses, it appeared that these dual biochemical properties originate from the existence of a certain variability in the β1-α1 loop. This allows positioning of several active site residues at proximity of the glutathione molecule, which itself remains unchanged in GSTF three-dimensional structures. These results highlight the promiscuity of some GSTFs and that changes of active site residues in some isoforms during evolution generated functional diversity by modifying their activity profile. Structural data are available in the PDB under the accession numbers 5EY6, 5F05, 5F06, and 5F07. © 2017 Federation of European Biochemical Societies.

  8. Identification of a highly reactive threonine residue at the active site of γ-glutamyl transpeptidase

    International Nuclear Information System (INIS)

    Stole, E.; Seddon, A.P.; Wellner, D.; Meister, A.

    1990-01-01

    γ-Glutamyl transpeptidase an enzyme of major importance in glutathione metabolism, was inactivated by treating it with L-(αS,5S)-α-amino-3-chloro-4,5-dihydro-5-[3- 14 C]isoxazoleacetic acid. This selective reagent binds stoichiometrically to the enzyme; more than 90% of the label was bound to its light subunit. Enzymatic digestion of the light subunit gave a 14 C-labeled peptide that corresponds to amino acid residues 517-527 of the enzyme and two incomplete digestion products that contain this labeled peptide moiety. The radioactivity associated with this peptide was released with threonine-523 during sequencing by the automated gas-phase Edman method. The light subunit contains 14 other threonine residues and a total of 19 serine residues; these were not labeled. Threonine-523 is situated in the enzyme in an environment that greatly increases its reactivity, indicating that other amino acid residues of the enzyme must also participate in the active-site chemistry of the enzyme

  9. Statistical modeling to management and treatment of scrap with low and very low residual activity

    International Nuclear Information System (INIS)

    Garcia-Bermejo Fernandez, R.; Anaya Lazaro, M.

    2011-01-01

    The experience of recent years on the management of scrap metal containing residual activity have allowed the development of a simple statistical model for the management of these materials. This statistical model includes a breakdown of the various processing operations to which these materials undergo and the effects in the process of radiological controls associated to the control of declassification that defines disposal (recycled by smelting, reclamation, temporary storage the plant or sent to final storage of radioactive waste.

  10. Chaperone-Like Activity of ß-Casein and Its Effect on Residual in Vitro Activity of Food Enzymes

    DEFF Research Database (Denmark)

    Sulewska, Anna Maria

    . The negative effect of BSA on enzyme was not observed before. The residual activity of horseradish peroxidase was also improved by the reconstituted skim milk: addition of reconstituted skim milk prior to heat treatment resulted in higher residual activity of HRP compared to no addition (58±3% and 30......±1%, respectively) The findings of this study show that β-casein can influence the response of food enzymes to heat treatment. β-Casein is not a universal chaperone and its effect on different targets needs to be evaluated on a case-by-case basis. This study also shows that proteins as e.g. BSA may affect......ABSTRACT Activity of endogenous enzymes may cause browning of fruits and vegetables. These enzymes can be inactivated, for example by heat treatment, but the response of enzymes to heat treatment depends on many factors. Foods are very complex systems and the stability of enzymes...

  11. Solid radioactive waste: evaluation of residual activity in nuclear medicine services

    International Nuclear Information System (INIS)

    Alabarse, Frederico G.; Xavier, Ana M.; Magalhaes, Maisa H.; Guerrero, Jesus S.P.

    2009-01-01

    An experimental programme to estimate, with a better degree of accuracy, the activity that remains adsorbed in flasks and syringes used in Nuclear Medicine Services for the administration of radionuclides to patients submitted to diagnostic or therapy is been conducted under the coordination of the Radioactive Waste Division of the Brazilian Nuclear Energy Commission, CNEN. The adopted recommendation in Brazil to allow an expedite solid waste management in nuclear medicine facilities, up to the present, is to consider that 2% of the initial activity remains adsorbed in the solid waste, which easily allows the calculation of the storage time to achieve regulatory clearance levels by decay. This research evaluates 17 different kinds of radiopharmaceuticals and three radioisotopes: 99m Tc, 67 Ga and 201 Tl. Results obtained by means of a weighting method to estimate the residual mass in flasks show that the ratio of the mass of the liquid that remains in the solid waste to the mass of the empty flask is constant. This suggests that the residual activity depends on the initial activity concentration of radiopharmaceutical contained in each flask, as assumed by the regulatory body. Additionally, results obtained by determining the remaining activity in flasks, shortly after the injection of its radionuclide contents in patients, indicate that an average value for the residual activity of the order of 10% of the initial activity contained in the flasks or syringes should be adopted to determine the decay storage time before the release of solid waste in the urban conventional land fill disposal system. The 'rule of thumb' of 10 half-lives for storage before clearance is also discussed in the present work. (author)

  12. Porcine reproductive and respiratory syndrome virus (PRRSV) up-regulates IL-8 expression through TAK-1/JNK/AP-1 pathways.

    Science.gov (United States)

    Liu, Yihao; Du, Yinping; Wang, Honglei; Du, Li; Feng, Wen-Hai

    2017-06-01

    The acute phase of respiratory distress caused by porcine reproductive and respiratory syndrome virus (PRRSV) is likely a consequence of the release of inflammatory cytokines in the lung. IL-8, the main chemokine and activator of neutrophils, might be related to the lung injury upon PRRSV infection. In this study, we showed that PRRSV induced IL-8 expression in vivo and in vitro. Subsequently, we demonstrated that JNK and NF-κB pathways were activated upon PRRSV infection and required for the enhancement of IL-8 expression. We further verified that PRRSV-activated TAK-1 was essential for the activation of JNK and NF-κB pathways and IL-8 expression. Moreover, we revealed an AP-1 binding motif in the cloned porcine IL-8 (pIL-8) promoter, and deletion of this motif abolished the pIL-8 promoter activity. Finally, we found that the JNK-activated AP-1 subunit c-Jun was critical for the up-regulation of IL-8 expression by PRRSV. These data suggest that PRRSV-induced IL-8 production is likely through the TAK-1/JNK/AP-1 pathways. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Relative toxicity and residual activity of insecticides used in blueberry pest management: mortality of natural enemies.

    Science.gov (United States)

    Roubos, Craig R; Rodriguez-Saona, Cesar; Holdcraft, Robert; Mason, Keith S; Isaacs, Rufus

    2014-02-01

    A series of bioassays were conducted to determine the relative toxicities and residual activities of insecticides labeled for use in blueberry (Vaccinium corymbosum L.) on natural enemies, to identify products with low toxicity or short duration effects on biological control agents. In total, 14 insecticides were evaluated using treated petri dishes and four commercially available natural enemies (Aphidius colemani Viereck, Orius insidiosus [Say], Chrysoperla rufilabris [Burmeister], and Hippodamia convergens [Guérin-Menéville]). Dishes were aged under greenhouse conditions for 0, 3, 7, or 14 d before introducing insects to test residual activity. Acute effects (combined mortality and knockdown) varied by insecticide, residue age, and natural enemy species. Broad-spectrum insecticides caused high mortality to all biocontrol agents, whereas products approved for use in organic agriculture had little effect. The reduced-risk insecticide acetamiprid consistently caused significant acute effects, even after aging for 14 d. Methoxyfenozide, novaluron, and chlorantraniliprole, which also are classified as reduced-risk insecticides, had low toxicity, and along with the organic products could be compatible with biological control. This study provides information to guide blueberry growers in their selection of insecticides. Further research will be needed to determine whether adoption of a pest management program based on the use of more selective insecticides will result in higher levels of biological control in blueberry.

  14. Review of the incineration of 500 tonnes of radio-active residues

    International Nuclear Information System (INIS)

    Rodier, J.; Seyfried, P.; Charbonneaux, M.

    1969-01-01

    During its first five years operation, the incinerator at the Marcoule Centre has burnt almost 500 tonnes of radio-active residues. Improvements in some of the details of the process have been made during this period; they concern the nature of the materials involved. The technical and radiological results for the installation are very favorable, and have made it possible to maintain a high charge factor.Although the overall economic results are not advantageous in the case of ungraded solid residues this method represents nevertheless the best available for eliminating oils, solvents, wood and dead animals. It can also be of use furthermore each time that a dilution in the atmosphere can advantageously be used as a method of disposing of certain radio elements such as tritium or carbon 14 in the form of gases or vapours. (author) [fr

  15. The masked cysteine residues in methylmalonyl-CoA mutase from Propionibacterium shermanii are essential for catalytic activity.

    Science.gov (United States)

    Roy, I

    1996-09-30

    Two masked cysteine residues have been reported in methylmalonyl-CoA mutase from Propionibacterium shermanii, Cys-535 in the alpha-subunit and Cys-517 in the beta-subunit, which are revealed only after reduction of the denatured enzyme with dithiothreitol. It has been postulated that these residues are involved in disulphide linkages to unknown thiols of low M(r). These two masked cysteine residues have been changed to an alanine, individually. Both the mutants, C535alphaA and C517betaA, were inactive. This shows that both these residues are essential for catalytic activity.

  16. Safety aspects of targets for ADTT: Activity, volatile products, residual heat release

    International Nuclear Information System (INIS)

    Gai, E.V.; Ignatyuk, A.V.; Lunev, V.P.; Shubin, Yu.N.

    1999-01-01

    Safety aspects of heavy metal liquid targets for the accelerator driven systems connected with the activity accumulation and residual energy release due to the irradiation with high energy proton beam are discussed. The results obtained for the lead-bismuth target that are under construction in IPPE now in the frame of ISTC Project No. 559 are briefly presented. The calculations and the analysis of the accumulation of the spallation reaction products, activity and energy release at various moments after the accelerator shutdown are presented. The concentrations of the reaction products, the total and partial activities, the activities of volatile products are determined. The contributions of the short-lived nuclides important for the prediction of the facility behaviour in regimes with the accelerator beam trips. The calculations and analysis of the residual energy release due to different decay type have been performed. The conclusions are as follows. The obtained results showed that long lived radioactivity accumulates mainly due to primary nuclear reactions. Secondary reactions are responsible for the production of small number of long-lived isotopes Bi-207, Po-210 and some others, being generated by radiative capture of low energy neutrons. It is possible to make a conclusion that neutrons in the energy range 20 - 800 MeV and protons with energy above 100 MeV give main contribution to the total activity generation although these parts of spectra inside the target give comparatively small contribution to the total flux. The correct consideration of short-lived nuclides contribution is the main problem in the analysis of the target behaviour in the case of short accelerator shutdowns. They make the determining contribution to the both activity and the heat release at the first moments after the accelerator shutdown, creating the intermediate links and additional channels for the long-lived nuclides accumulation chains. The strong dependence of calculated

  17. Residual activity evaluation: a benchmark between ANITA, FISPACT, FLUKA and PHITS codes

    Science.gov (United States)

    Firpo, Gabriele; Viberti, Carlo Maria; Ferrari, Anna; Frisoni, Manuela

    2017-09-01

    The activity of residual nuclides dictates the radiation fields in periodic inspections/repairs (maintenance periods) and dismantling operations (decommissioning phase) of accelerator facilities (i.e., medical, industrial, research) and nuclear reactors. Therefore, the correct prediction of the material activation allows for a more accurate planning of the activities, in line with the ALARA (As Low As Reasonably Achievable) principles. The scope of the present work is to show the results of a comparison between residual total specific activity versus a set of cooling time instants (from zero up to 10 years after irradiation) as obtained by two analytical (FISPACT and ANITA) and two Monte Carlo (FLUKA and PHITS) codes, making use of their default nuclear data libraries. A set of 40 irradiating scenarios is considered, i.e. neutron and proton particles of different energies, ranging from zero to many hundreds MeV, impinging on pure elements or materials of standard composition typically used in industrial applications (namely, AISI SS316 and Portland concrete). In some cases, experimental results were also available for a more thorough benchmark.

  18. Residual activity evaluation: a benchmark between ANITA, FISPACT, FLUKA and PHITS codes

    Directory of Open Access Journals (Sweden)

    Firpo Gabriele

    2017-01-01

    Full Text Available The activity of residual nuclides dictates the radiation fields in periodic inspections/repairs (maintenance periods and dismantling operations (decommissioning phase of accelerator facilities (i.e., medical, industrial, research and nuclear reactors. Therefore, the correct prediction of the material activation allows for a more accurate planning of the activities, in line with the ALARA (As Low As Reasonably Achievable principles. The scope of the present work is to show the results of a comparison between residual total specific activity versus a set of cooling time instants (from zero up to 10 years after irradiation as obtained by two analytical (FISPACT and ANITA and two Monte Carlo (FLUKA and PHITS codes, making use of their default nuclear data libraries. A set of ~40 irradiating scenarios is considered, i.e. neutron and proton particles of different energies, ranging from zero to many hundreds MeV, impinging on pure elements or materials of standard composition typically used in industrial applications (namely, AISI SS316 and Portland concrete. In some cases, experimental results were also available for a more thorough benchmark.

  19. Ventricular activity cancellation in electrograms during atrial fibrillation with constraints on residuals' power.

    Science.gov (United States)

    Corino, Valentina D A; Rivolta, Massimo W; Sassi, Roberto; Lombardi, Federico; Mainardi, Luca T

    2013-12-01

    During atrial fibrillation (AF), cancellation of ventricular activity from atrial electrograms (AEG) is commonly performed by template matching and subtraction (TMS): a running template, built in correspondence of QRSs, is subtracted from the AEG to uncover atrial activity (AA). However, TMS can produce poor cancellation, leaving high-power residues. In this study, we propose to modulate the templates before subtraction, in order to make the residuals as similar as possible to the nearby atrial activity, avoiding high-power ones. The coefficients used to modulate the template are estimated by maximizing, via Multi-swarm Particle Swarm Optimization, a fitness function. The modulated TMS method (mTMS) was tested on synthetic and real AEGs. Cancellation performances were assessed using: normalized mean squared error (NMSE, computed on simulated data only), reduction of ventricular activity (VDR), and percentage of segments (PP) whose power was outside the standard range of the atrial power. All testings suggested that mTMS is an improvement over TMS alone, being, on simulated data, NMSE and PP significantly decreased while VDR significantly increased. Similar results were obtained on real electrograms (median values of CS1 recordings PP: 2.44 vs. 0.38 p < 0.001; VDR: 6.71 vs. 8.15 p < 0.001). Copyright © 2013 IPEM. Published by Elsevier Ltd. All rights reserved.

  20. The effects of residual platelets in plasma on plasminogen activator inhibitor-1 and plasminogen activator inhibitor-1-related assays.

    Directory of Open Access Journals (Sweden)

    Marlien Pieters

    Full Text Available Due to controversial evidence in the literature pertaining to the activity of plasminogen activator inhibitor-1 in platelets, we examined the effects of residual platelets present in plasma (a potential pre-analytical variable on various plasminogen activator inhibitor-1 and plasminogen activator inhibitor-1-related assays. Blood samples were collected from 151 individuals and centrifuged at 352 and 1500 g to obtain plasma with varying numbers of platelet. In a follow-up study, blood samples were collected from an additional 23 individuals, from whom platelet-poor (2000 g, platelet-containing (352 g and platelet-rich plasma (200 g were prepared and analysed as fresh-frozen and after five defrost-refreeze cycles (to determine the contribution of in vitro platelet degradation. Plasminogen activator inhibitor-1 activity, plasminogen activator inhibitor-1 antigen, tissue plasminogen activator/plasminogen activator inhibitor-1 complex, plasma clot lysis time, β-thromboglobulin and plasma platelet count were analysed. Platelet α-granule release (plasma β-thromboglobulin showed a significant association with plasminogen activator inhibitor-1 antigen levels but weak associations with plasminogen activator inhibitor-1 activity and a functional marker of fibrinolysis, clot lysis time. Upon dividing the study population into quartiles based on β-thromboglobulin levels, plasminogen activator inhibitor-1 antigen increased significantly across the quartiles while plasminogen activator inhibitor-1 activity and clot lysis time tended to increase in the 4th quartile only. In the follow-up study, plasma plasminogen activator inhibitor-1 antigen was also significantly influenced by platelet count in a concentration-dependent manner. Plasma plasminogen activator inhibitor-1 antigen levels increased further after complete platelet degradation. Residual platelets in plasma significantly influence plasma plasminogen activator inhibitor-1 antigen levels mainly

  1. Identification of Residues in the Lipopolysaccharide ABC Transporter That Coordinate ATPase Activity with Extractor Function.

    Science.gov (United States)

    Simpson, Brent W; Owens, Tristan W; Orabella, Matthew J; Davis, Rebecca M; May, Janine M; Trauger, Sunia A; Kahne, Daniel; Ruiz, Natividad

    2016-10-18

    The surface of most Gram-negative bacteria is covered with lipopolysaccharide (LPS), creating a permeability barrier against toxic molecules, including many antimicrobials. To assemble LPS on their surface, Gram-negative bacteria must extract newly synthesized LPS from the inner membrane, transport it across the aqueous periplasm, and translocate it across the outer membrane. The LptA to -G proteins assemble into a transenvelope complex that transports LPS from the inner membrane to the cell surface. The Lpt system powers LPS transport from the inner membrane by using a poorly characterized ATP-binding cassette system composed of the ATPase LptB and the transmembrane domains LptFG. Here, we characterize a cluster of residues in the groove region of LptB that is important for controlling LPS transport. We also provide the first functional characterization of LptFG and identify their coupling helices that interact with the LptB groove. Substitutions at conserved residues in these coupling helices compromise both the assembly and function of the LptB 2 FG complex. Defects in LPS transport conferred by alterations in the LptFG coupling helices can be rescued by changing a residue in LptB that is adjacent to functionally important residues in the groove region. This suppression is achieved by increasing the ATPase activity of the LptB 2 FG complex. Taken together, these data identify a specific binding site in LptB for the coupling helices of LptFG that is responsible for coupling of ATP hydrolysis by LptB with LptFG function to achieve LPS extraction. Lipopolysaccharide (LPS) is synthesized at the cytoplasmic membrane of Gram-negative bacteria and transported across several compartments to the cell surface, where it forms a barrier that protects these organisms from antibiotics. The LptB 2 FG proteins form an ATP-binding cassette (ABC) transporter that uses energy from ATP hydrolysis in the cytoplasm to facilitate extraction of LPS from the outer face of the

  2. ZnCl{sub 2}-activated biochar from biogas residue facilitates aqueous As(III) removal

    Energy Technology Data Exchange (ETDEWEB)

    Xia, Dong; Tan, Fen; Zhang, Chuanpan [Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, and The Key Laboratory for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen 361005 (China); Jiang, Xiuli; Chen, Zheng; Li, Heng [Environmental Science Research Center, College of the Environment & Ecology, Xiamen University, Xiamen 361110 (China); Zheng, Yanmei [Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, and The Key Laboratory for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen 361005 (China); Li, Qingbiao [Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, and The Key Laboratory for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen 361005 (China); Environmental Science Research Center, College of the Environment & Ecology, Xiamen University, Xiamen 361110 (China); Wang, Yuanpeng, E-mail: wypp@xmu.edu.cn [Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, and The Key Laboratory for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen 361005 (China)

    2016-07-30

    Highlights: • The ZnCl{sub 2}-activated biochar from the biogas residue of pig manure showed an excellent ability to remove As(III). • ZnCl{sub 2}-activated biochar had a large BET surface area and well-distributed pore structure. • Zinc played a dominant role in the removal of As(III) by forming Zn-O-As(III). - Abstract: Biochars prepared from biogas residue using different chemical activators were investigated for their As(III) adsorption properties. The results indicated that the original biochars did not exhibit significant As(III) adsorption. However, ZnCl{sub 2}-activated biochar, which possessed the largest specific surface area, 516.67 cm{sup 2}/g, and exhibited a perfectly porous texture, showed excellent performance in a 500 μgL{sup −1} solution of As(III). Fourier transform infrared spectroscopy (FT-IR) and X-ray photoelectron spectroscopy were utilized to identify the mechanism of As(III) adsorption by ZnCl{sub 2}-activated biochar. Adsorption was found to occur mainly through ligand exchange of the hydroxyl in Zn-OH to form Zn-O-As(III), as well as through porous adsorption. As a low-cost adsorbent, the adsorption process was well fitted using a pseudo-second-order model, with an R{sup 2} > 0.993. The adsorption process was fast, requiring nearly 90 min to reach adsorption equilibrium. Batch adsorption experimental results indicated that ZnCl{sub 2}-activated biochar has a maximum adsorption capacity of 27.67 mg/g at pH 7.0, and the adsorption process followed the Freundlich isotherm model well, with an R{sup 2} > 0.994. In addition, the current work demonstrated the efficiency of using ZnCl{sub 2}-activated biochar adsorbent to treat As(III)-contaminated water.

  3. Phenol Adsorption on Nitrogen-enriched Activated Carbon Prepared from Bamboo Residues

    OpenAIRE

    Ji Zhang; Xiao-Juan Jin; Jian-Min Gao; Xiu-Dong Zhang

    2013-01-01

    Nitrogen-enriched activated carbons prepared from bamboo residues were characterized by means of BET, XPS, and elemental analysis. Then adsorption experiments were carried out to study the effects of various physicochemical parameters such as contact time, temperature, pH, and initial concentration. Adsorption equilibrium was achieved within 120 min at a phenol concentration of 250 mg/L. When the pH was 4 and 0.1 g of the carbon absorbent and 100 mL of phenol solution at 250 mg/L were used, t...

  4. C-H activation on rhodium: reaction mechanism and the role of carbonaceous residues

    International Nuclear Information System (INIS)

    Cogen, J.M.; Maier, W.F.

    1986-01-01

    Evidence for two C-H activation mechanisms, the oxidative addition of a single C-H bond and the concerted oxidative addition of two adjacent C-H bonds, is presented. The two mechanisms show different sensitivity to surface structure or particle size. While the first process (E/sub a/ ∼ 21 kcal/mol) is insensitive to the hydrocarbon structure, the second process (E/sub a/ ∼ 5-9 kcal/mol) is sensitive to the hydrocarbon structure. C-H activation is found to be 0.6 order in hexane and zero order in deuterium, which is consistent with oxidative addition as the rate-determining step. The d 2 maximum obtained by H/D exchange of linear alkanes on Rh results from surface olefins, as indicated by high-resolution deuterium NMR. Evidence against the participation of carbonaceous residues in the H/D exchange process is presented, indicating that carbonaceous material is not part of the active site. Two types of carbonaceous residues are detected. One forms at lower temperatures and does not interfere with the reaction; the other forms at higher temperatures and acts to poison the catalyst

  5. Residual activity induced by heavy ions and beam-loss criteria for heavy-ion accelerators

    Directory of Open Access Journals (Sweden)

    I. Strašík

    2010-07-01

    Full Text Available The paper presents results of FLUKA simulations of the residual activity induced by heavy ions in two target configurations representing: (1 a beam pipe of an accelerator and (2 a bulky accelerator structure like a magnet yoke or a coil. The target materials were stainless steel and copper representing the most common construction materials used for basic accelerator components. For these two materials, the inventory of the induced isotopes depends mainly on the target material and much less on the projectile species. Time evolution of the induced activity can be described by means of a generic curve that is independent from the projectile mass. Dependence of the induced residual activity on selected ion beam parameters was studied. The main goal of the study was establishing a scaling law expanding the existing proton beam-loss tolerance to heavy-ion beams. This scaling law enables specifying beam-loss criteria for projectile species from proton up to uranium at energies from 200  MeV/u up to 1  GeV/u.

  6. Tripartite Motif 8 (TRIM8) Positively Regulates Pro-inflammatory Responses in Pseudomonas aeruginosa-Induced Keratitis Through Promoting K63-Linked Polyubiquitination of TAK1 Protein.

    Science.gov (United States)

    Guo, Litao; Dong, Weili; Fu, Xiaoxiao; Lin, Jing; Dong, Zhijun; Tan, Xiaobo; Zhang, Tiemin

    2017-04-01

    Pseudomonas aeruginosa (PA)-induced keratitis is a rapidly progressive ocular infectious disease that often leads to inflammatory epithelial edema, stromal infiltration, tissue destruction, corneal ulceration, and vision loss. In this study, we investigate the role of tripartite motif 8 (TRIM8) in regulating the inflammatory process of PA-induced keratitis. The expression of TRIM8 was increased in mouse corneas and in vitro-cultured macrophages after PA infection. Knockdown of the expression of TRIM8 significantly inhibited the activation of NF-κB signaling and decreased the production of pro-inflammatory cytokines both in vivo and in vitro after infected with PA. Furthermore, we investigated the potential mechanism and we found after PA infection that TRIM8 could promote K63-linked polyubiquitination of transforming growth factor β-activated kinase 1 (TAK1), leading to the activation of TAK1 and enhanced inflammatory responses. Taken together, we demonstrated that TRIM8 has pro-inflammatory effect on PA-induced keratitis and suggest TRIM8 as a potential therapeutic target for keratitis.

  7. TAK-1/p38/nNFκB signaling inhibits myoblast differentiation by increasing levels of Activin A

    Directory of Open Access Journals (Sweden)

    Trendelenburg Anne

    2012-02-01

    Full Text Available Abstract Background Skeletal-muscle differentiation is required for the regeneration of myofibers after injury. The differentiation capacity of satellite cells is impaired in settings of old age, which is at least one factor in the onset of sarcopenia, the age-related loss of skeletal-muscle mass and major cause of frailty. One important cause of impaired regeneration is increased levels of transforming growth factor (TGF-β accompanied by reduced Notch signaling. Pro-inflammatory cytokines are also upregulated in aging, which led us hypothesize that they might potentially contribute to impaired regeneration in sarcopenia. Thus, in this study, we further analyzed the muscle differentiation-inhibition pathway mediated by pro-inflammatory cytokines in human skeletal muscle cells (HuSKMCs. Methods We studied the modulation of HuSKMC differentiation by the pro-inflammatory cytokines interleukin (IL-1α and tumor necrosis factor (TNF-α The grade of differentiation was determined by either imaging (fusion index or creatine kinase (CK activity, a marker of muscle differentiation. Secretion of TGF-β proteins during differentiation was assessed by using a TGF-β-responsive reporter-gene assay and further identified by means of pharmacological and genetic inhibitors. In addition, signaling events were monitored by western blotting and reverse transcription PCR, both in HuSKMC cultures and in samples from a rat sarcopenia study. Results The pro-inflammatory cytokines IL-1α and TNF-α block differentiation of human myoblasts into myotubes. This anti-differentiation effect requires activation of TGF-β-activated kinase (TAK-1. Using pharmacological and genetic inhibitors, the TAK-1 pathway could be traced to p38 and NFκB. Surprisingly, the anti-differentiation effect of the cytokines required the transcriptional upregulation of Activin A, which in turn acted through its established signaling pathway: ActRII/ALK/SMAD. Inhibition of Activin A signaling was

  8. Activation of the LRR Receptor-Like Kinase PSY1R Requires Transphosphorylation of Residues in the Activation Loop

    Directory of Open Access Journals (Sweden)

    Christian B. Oehlenschlæger

    2017-11-01

    Full Text Available PSY1R is a leucine-rich repeat (LRR receptor-like kinase (RLK previously shown to act as receptor for the plant peptide hormone PSY1 (peptide containing sulfated tyrosine 1 and to regulate cell expansion. PSY1R phosphorylates and thereby regulates the activity of plasma membrane-localized H+-ATPases. While this mechanism has been studied in detail, little is known about how PSY1R itself is activated. Here we studied the activation mechanism of PSY1R. We show that full-length PSY1R interacts with members of the SERK co-receptor family in planta. We identified seven in vitro autophosphorylation sites on serine and threonine residues within the kinase domain of PSY1R using mass spectrometry. We furthermore show that PSY1R autophosphorylation occurs in trans and that the initial transphosphorylation takes place within the activation loop at residues Ser951, Thr959, and Thr963. While Thr959 and Thr963 are conserved among other related plant LRR RLKs, Ser951 is unique to PSY1R. Based on homology modeling we propose that phosphorylation of Ser951 stabilize the inactive conformation of PSY1R.

  9. Activity and Residual Effect of Two Formulations of Lambdacyhalothrin Sprayed on Palm Leaves to Rhodnius prolixus

    Directory of Open Access Journals (Sweden)

    Mazariego-Arana Miguel Angel

    2002-01-01

    Full Text Available The insecticidal activity and residual effect of two formulations of lambdacyhalothrin were evaluated with Rhodnius prolixus;laboratory and field tests were conducted in the State of Chiapas, Mexico. The results indicate that the lethal concentrations of the active ingredient of SC (LC50 = 2.37 and LC90 = 8.5 mg, a.i./m² were 4-8 times than those with the insecticide WP applied on R. prolixus bugs in palm leaves, a common building material for thatched roofs. Other investigators in South America recommended applying 30 mg a.i./m² in porous materials; we obtained that the products WP and SC were 3.5 and 16 times more effective on palm leaves. Regarding the evaluation of the residual effects in field spraying, there was up to 15 months persistence after the application of WP in two doses (8.6 mg a.i./m² and 3.752 mg a.i./m² with SC. We consider R. prolixus highly susceptible to the employed pyrethroids; they could be used to control this vector in the state of Chiapas, Mexico.

  10. Identification of key residues that confer Rhodobacter sphaeroides LPS activity at horse TLR4/MD-2.

    Directory of Open Access Journals (Sweden)

    Katherine L Irvine

    Full Text Available The molecular determinants underpinning how hexaacylated lipid A and tetraacylated precursor lipid IVa activate Toll-like receptor 4 (TLR4 are well understood, but how activation is induced by other lipid A species is less clear. Species specificity studies have clarified how TLR4/MD-2 recognises different lipid A structures, for example tetraacylated lipid IVa requires direct electrostatic interactions for agonism. In this study, we examine how pentaacylated lipopolysaccharide from Rhodobacter sphaeroides (RSLPS antagonises human TLR4/MD-2 and activates the horse receptor complex using a computational approach and cross-species mutagenesis. At a functional level, we show that RSLPS is a partial agonist at horse TLR4/MD-2 with greater efficacy than lipid IVa. These data suggest the importance of the additional acyl chain in RSLPS signalling. Based on docking analysis, we propose a model for positioning of the RSLPS lipid A moiety (RSLA within the MD-2 cavity at the TLR4 dimer interface, which allows activity at the horse receptor complex. As for lipid IVa, RSLPS agonism requires species-specific contacts with MD-2 and TLR4, but the R2 chain of RSLA protrudes from the MD-2 pocket to contact the TLR4 dimer in the vicinity of proline 442. Our model explains why RSLPS is only partially dependent on horse TLR4 residue R385, unlike lipid IVa. Mutagenesis of proline 442 into a serine residue, as found in human TLR4, uncovers the importance of this site in RSLPS signalling; horse TLR4 R385G/P442S double mutation completely abolishes RSLPS activity without its counterpart, human TLR4 G384R/S441P, being able to restore it. Our data highlight the importance of subtle changes in ligand positioning, and suggest that TLR4 and MD-2 residues that may not participate directly in ligand binding can determine the signalling outcome of a given ligand. This indicates a cooperative binding mechanism within the receptor complex, which is becoming increasingly

  11. The arginine residue within the C-terminal active core of Bombyx mori pheromone biosynthesis-activating neuropeptide (PBAN is essential for receptor binding and activation

    Directory of Open Access Journals (Sweden)

    Takeshi eKawai

    2012-03-01

    Full Text Available In most lepidopteran insects, the biosynthesis of sex pheromones is regulated by pheromone biosynthesis activating neuropeptide (PBAN. Bombyx mori PBAN (BomPBAN consists of 33 amino acid residues and contains a C-terminus FSPRLamide motif as the active core. Among neuropeptides containing the FXPRLamide motif, the arginine (Arg, R residue two positions from the C-terminus is highly conserved across several neuropeptides, which can be designated as RXamide peptides. The purpose of this study was to reveal the role of the Arg residue in the BomPBAN active core. We synthesized a ten-residue peptide corresponding to the C-terminal part of BomPBAN with a series of point mutants at the 2nd position (ie, Arg from the C-terminus, termed the C2 position, and measured their efficacy in stimulating Ca2+ influx in insect cells concomitantly expressing a fluorescent PBAN receptor chimera (PBANR-EGFP and loaded with the fluorescent Ca2+ indicator, Fura Red-AM. PBAN analogs with the C2 position replaced with alanine (Ala, A, aspartic acid (Asp, D, serine (Ser, S or L-2-aminooctanoic acid (Aoc decreased PBAN-like activity. RC2A (SKTRYFSPALamide and RC2D (SKTRYFSPDLamide had the lowest activity and could not inhibit the activity of PBAN C10 (SKTRYFSPRLamide. We also prepared Rhodamine Red-labeled PBAN analogs of the mutants and examined their ability to bind PBANR. In contrast to 100 nM Rhodamine Red-PBAN C10, none of the mutants at the same concentration exhibited PBANR binding. Taken together, our results demonstrate that the C2 Arg residue in BomPBAN is essential for PBANR binding and activation.

  12. Effects of non-catalytic, distal amino acid residues on activity of E. coli DinB (DNA polymerase IV).

    Science.gov (United States)

    Walsh, Jason M; Parasuram, Ramya; Rajput, Pradyumna R; Rozners, Eriks; Ondrechen, Mary Jo; Beuning, Penny J

    2012-12-01

    DinB is one of two Y family polymerases in E. coli and is involved in copying damaged DNA. DinB is specialized to bypass deoxyguanosine adducts that occur at the N(2) position, with its cognate lesion being the furfuryl adduct. Active site residues have been identified that make contact with the substrate and carry out deoxynucleotide triphosphate (dNTP) addition to the growing DNA strand. In DNA polymerases, these include negatively charged aspartate and glutamate residues (D8, D103, and E104 in E. coli DNA polymerase IV DinB). These residues position the essential magnesium ions correctly to facilitate nucleophilic attack by the primer hydroxyl group on the α-phosphate group of the incoming dNTP. To study the contribution of DinB residues to lesion bypass, the computational methods THEMATICS and POOL were employed. These methods correctly predict the known active site residues, as well as other residues known to be important for activity. In addition, these methods predict other residues involved in substrate binding as well as more remote residues. DinB variants with mutations at the predicted positions were constructed and assayed for bypass of the N(2) -furfuryl-dG lesion. We find a wide range of effects of predicted residues, including some mutations that abolish damage bypass. Moreover, most of the DinB variants constructed are unable to carry out the extension step of lesion bypass. The use of computational prediction methods represents another tool that will lead to a more complete understanding of translesion DNA synthesis. Copyright © 2012 Wiley Periodicals, Inc.

  13. Characterization of the heterotrophic biomass and the endogenous residue of activated sludge.

    Science.gov (United States)

    Ramdani, Abdellah; Dold, Peter; Gadbois, Alain; Déléris, Stéphane; Houweling, Dwight; Comeau, Yves

    2012-03-01

    The activated sludge process generates an endogenous residue (X(E)) as a result of heterotrophic biomass decay (X(H)). A literature review yielded limited information on the differences between X(E) and X(H) in terms of chemical composition and content of extracellular polymeric substances (EPS). The objective of this project was to characterize the chemical composition (x, y, z, a, b and c in C(x)H(y)O(z)N(a)P(b)S(c)) of the endogenous and the active fractions and EPS of activated sludge from well designed experiments. To isolate X(H) and X(E) in this study, activated sludge was generated in a 200L pilot-scale aerobic membrane bioreactor (MBR) fed with a soluble and completely biodegradable synthetic influent of sodium acetate as the sole carbon source. This influent, which contained no influent unbiodegradable organic or inorganic particulate matter, allowed the generation of a sludge composed essentially of two fractions: heterotrophic biomass X(H) and an endogenous residue X(E), the nitrifying biomass being negligible. The endogenous decay rate and the active biomass fraction of the MBR sludge were determined in 21-day aerobic digestion batch tests by monitoring the VSS and OUR responses. Fractions of X(H) and X(E) were respectively 68% and 32% in run 1 (MBR at 5.2 day SRT) and 59% and 41% in run 2 (MBR at 10.4 day SRT). The endogenous residue was isolated by subjecting the MBR sludge to prolonged aerobic batch digestion for 3 weeks, and was characterized in terms of (a) elemental analysis for carbon, nitrogen, phosphorus and sulphur; and (b) content of EPS. The MBR sludge was characterized using the same procedures (a and b). Knowing the proportions of X(H) and X(E) in this sludge, it was possible to characterize X(H) by back calculation. Results from this investigation showed that the endogenous residue had a chemical composition different from that of the active biomass with a lower content of inorganic matter (1:4.2), of nitrogen (1:2.9), of phosphorus (1

  14. Application of Scharer's quantitative method for the determination of residual alkaline phosphatase activity in standard Minas

    Directory of Open Access Journals (Sweden)

    C.F. Soares

    2013-08-01

    Full Text Available Milk pasteurization is a critical issue in the dairy industry, and failures in this process can affect final product safety. Scharer's enzymatic method is still traditionally used to verify pasteurization efficiency compliance, and it is based on screening for residual alkaline phosphatase in milk. Although several methods are used to quantify enzymatic activity to assess milk pasteurization efficiency, there is a small amount of published data regarding the use of these methods to quantify alkaline phosphatase in cheese. In this study, the Scharer's modified method was used to determine the levels of residual alkaline phosphatase in standard minas cheese, before and after 20 days of ripening. The cheeses were made using raw or pasteurized milk with the addition of different concentrations of raw milk (0; 0.05%; 0.10%; 0.20%; and 0.50%. In the fresh cheese samples, the method showed a sensitivity of only 0.50% with the addition of raw milk to the pasteurized milk used to make cheese. In addition, levels of up 0.20% of raw milk in pasteurized milk, the concentrations of phenol was inferior to 1μg phenol/g of dairy product which is the preconized indicator value for adequate pasteurization.

  15. Procedure of Active Residual Heat Removal after Emergency Shutdown of High-Temperature-Gas-Cooled Reactor

    Directory of Open Access Journals (Sweden)

    Xingtuan Yang

    2014-01-01

    Full Text Available After emergency shutdown of high-temperature-gas-cooled reactor, the residual heat of the reactor core should be removed. As the natural circulation process spends too long period of time to be utilized, an active residual heat removal procedure is needed, which makes use of steam generator and start-up loop. During this procedure, the structure of steam generator may suffer cold/heat shock because of the sudden load of coolant or hot helium at the first few minutes. Transient analysis was carried out based on a one-dimensional mathematical model for steam generator and steam pipe of start-up loop to achieve safety and reliability. The results show that steam generator should be discharged and precooled; otherwise, boiling will arise and introduce a cold shock to the boiling tubes and tube sheet when coolant began to circulate prior to the helium. Additionally, in avoiding heat shock caused by the sudden load of helium, the helium circulation should be restricted to start with an extreme low flow rate; meanwhile, the coolant of steam generator (water should have flow rate as large as possible. Finally, a four-step procedure with precooling process of steam generator was recommended; sensitive study for the main parameters was conducted.

  16. Human xanthine oxidase changes its substrate specificity to aldehyde oxidase type upon mutation of amino acid residues in the active site: roles of active site residues in binding and activation of purine substrate.

    Science.gov (United States)

    Yamaguchi, Yuichiro; Matsumura, Tomohiro; Ichida, Kimiyoshi; Okamoto, Ken; Nishino, Takeshi

    2007-04-01

    Xanthine oxidase (oxidoreductase; XOR) and aldehyde oxidase (AO) are similar in protein structure and prosthetic group composition, but differ in substrate preference. Here we show that mutation of two amino acid residues in the active site of human XOR for purine substrates results in conversion of the substrate preference to AO type. Human XOR and its Glu803-to-valine (E803V) and Arg881-to-methionine (R881M) mutants were expressed in an Escherichia coli system. The E803V mutation almost completely abrogated the activity towards hypoxanthine as a substrate, but very weak activity towards xanthine remained. On the other hand, the R881M mutant lacked activity towards xanthine, but retained slight activity towards hypoxanthine. Both mutants, however, exhibited significant aldehyde oxidase activity. The crystal structure of E803V mutant of human XOR was determined at 2.6 A resolution. The overall molybdopterin domain structure of this mutant closely resembles that of bovine milk XOR; amino acid residues in the active centre pocket are situated at very similar positions and in similar orientations, except that Glu803 was replaced by valine, indicating that the decrease in activity towards purine substrate is not due to large conformational change in the mutant enzyme. Unlike wild-type XOR, the mutants were not subject to time-dependent inhibition by allopurinol.

  17. Mutational analysis of GlnB residues critical for NifA activation in Azospirillum brasilense.

    Science.gov (United States)

    Inaba, Juliana; Thornton, Jeremy; Huergo, Luciano Fernandes; Monteiro, Rose Adele; Klassen, Giseli; Pedrosa, Fábio de Oliveira; Merrick, Mike; de Souza, Emanuel Maltempi

    2015-02-01

    PII proteins are signal transduction that sense cellular nitrogen status and relay this signals to other targets. Azospirillum brasilense is a nitrogen fixing bacterium, which associates with grasses and cereals promoting beneficial effects on plant growth and crop yields. A. brasilense contains two PII encoding genes, named glnB and glnZ. In this paper, glnB was mutagenised in order to identify amino acid residues involved in GlnB signaling. Two variants were obtained by random mutagenesis, GlnBL13P and GlnBV100A and a site directed mutant, GlnBY51F, was obtained. Their ability to complement nitrogenase activity of glnB mutant strains of A. brasilense were determined. The variant proteins were also overexpressed in Escherichia coli, purified and characterized biochemically. None of the GlnB variant forms was able to restore nitrogenase activity in glnB mutant strains of A. brasilense LFH3 and 7628. The purified GlnBY51F and GlnBL13P proteins could not be uridylylated by GlnD, whereas GlnBV100A was uridylylated but at only 20% of the rate for wild type GlnB. Biochemical and computational analyses suggest that residue Leu13, located in the α helix 1 of GlnB, is important to maintain GlnB trimeric structure and function. The substitution V100A led to a lower affinity for ATP binding. Together the results suggest that NifA activation requires uridylylated GlnB bound to ATP. Copyright © 2014 Elsevier GmbH. All rights reserved.

  18. Muscle activation patterns during walking from transtibial amputees recorded within the residual limb-prosthetic interface

    Science.gov (United States)

    2012-01-01

    Background Powered lower limb prostheses could be more functional if they had access to feedforward control signals from the user’s nervous system. Myoelectric signals are one potential control source. The purpose of this study was to determine if muscle activation signals could be recorded from residual lower limb muscles within the prosthetic socket-limb interface during walking. Methods We recorded surface electromyography from three lower leg muscles (tibilias anterior, gastrocnemius medial head, gastrocnemius lateral head) and four upper leg muscles (vastus lateralis, rectus femoris, biceps femoris, and gluteus medius) of 12 unilateral transtibial amputee subjects and 12 non-amputee subjects during treadmill walking at 0.7, 1.0, 1.3, and 1.6 m/s. Muscle signals were recorded from the amputated leg of amputee subjects and the right leg of control subjects. For amputee subjects, lower leg muscle signals were recorded from within the limb-socket interface and from muscles above the knee. We quantified differences in the muscle activation profile between amputee and control groups during treadmill walking using cross-correlation analyses. We also assessed the step-to-step inter-subject variability of these profiles by calculating variance-to-signal ratios. Results We found that amputee subjects demonstrated reliable muscle recruitment signals from residual lower leg muscles recorded within the prosthetic socket during walking, which were locked to particular phases of the gait cycle. However, muscle activation profile variability was higher for amputee subjects than for control subjects. Conclusion Robotic lower limb prostheses could use myoelectric signals recorded from surface electrodes within the socket-limb interface to derive feedforward commands from the amputee’s nervous system. PMID:22882763

  19. Muscle activation patterns during walking from transtibial amputees recorded within the residual limb-prosthetic interface

    Directory of Open Access Journals (Sweden)

    Huang Stephanie

    2012-08-01

    Full Text Available Abstract Background Powered lower limb prostheses could be more functional if they had access to feedforward control signals from the user’s nervous system. Myoelectric signals are one potential control source. The purpose of this study was to determine if muscle activation signals could be recorded from residual lower limb muscles within the prosthetic socket-limb interface during walking. Methods We recorded surface electromyography from three lower leg muscles (tibilias anterior, gastrocnemius medial head, gastrocnemius lateral head and four upper leg muscles (vastus lateralis, rectus femoris, biceps femoris, and gluteus medius of 12 unilateral transtibial amputee subjects and 12 non-amputee subjects during treadmill walking at 0.7, 1.0, 1.3, and 1.6 m/s. Muscle signals were recorded from the amputated leg of amputee subjects and the right leg of control subjects. For amputee subjects, lower leg muscle signals were recorded from within the limb-socket interface and from muscles above the knee. We quantified differences in the muscle activation profile between amputee and control groups during treadmill walking using cross-correlation analyses. We also assessed the step-to-step inter-subject variability of these profiles by calculating variance-to-signal ratios. Results We found that amputee subjects demonstrated reliable muscle recruitment signals from residual lower leg muscles recorded within the prosthetic socket during walking, which were locked to particular phases of the gait cycle. However, muscle activation profile variability was higher for amputee subjects than for control subjects. Conclusion Robotic lower limb prostheses could use myoelectric signals recorded from surface electrodes within the socket-limb interface to derive feedforward commands from the amputee’s nervous system.

  20. Muscle activation patterns during walking from transtibial amputees recorded within the residual limb-prosthetic interface.

    Science.gov (United States)

    Huang, Stephanie; Ferris, Daniel P

    2012-08-10

    Powered lower limb prostheses could be more functional if they had access to feedforward control signals from the user's nervous system. Myoelectric signals are one potential control source. The purpose of this study was to determine if muscle activation signals could be recorded from residual lower limb muscles within the prosthetic socket-limb interface during walking. We recorded surface electromyography from three lower leg muscles (tibilias anterior, gastrocnemius medial head, gastrocnemius lateral head) and four upper leg muscles (vastus lateralis, rectus femoris, biceps femoris, and gluteus medius) of 12 unilateral transtibial amputee subjects and 12 non-amputee subjects during treadmill walking at 0.7, 1.0, 1.3, and 1.6 m/s. Muscle signals were recorded from the amputated leg of amputee subjects and the right leg of control subjects. For amputee subjects, lower leg muscle signals were recorded from within the limb-socket interface and from muscles above the knee. We quantified differences in the muscle activation profile between amputee and control groups during treadmill walking using cross-correlation analyses. We also assessed the step-to-step inter-subject variability of these profiles by calculating variance-to-signal ratios. We found that amputee subjects demonstrated reliable muscle recruitment signals from residual lower leg muscles recorded within the prosthetic socket during walking, which were locked to particular phases of the gait cycle. However, muscle activation profile variability was higher for amputee subjects than for control subjects. Robotic lower limb prostheses could use myoelectric signals recorded from surface electrodes within the socket-limb interface to derive feedforward commands from the amputee's nervous system.

  1. Identification of Residues Controlling Restriction versus Enhancing Activities of IFITM Proteins on Entry of Human Coronaviruses.

    Science.gov (United States)

    Zhao, Xuesen; Sehgal, Mohit; Hou, Zhifei; Cheng, Junjun; Shu, Sainan; Wu, Shuo; Guo, Fang; Le Marchand, Sylvain J; Lin, Hanxin; Chang, Jinhong; Guo, Ju-Tao

    2018-03-15

    Interferon-induced transmembrane proteins (IFITMs) are restriction factors that inhibit the infectious entry of many enveloped RNA viruses. However, we demonstrated previously that human IFITM2 and IFITM3 are essential host factors facilitating the entry of human coronavirus (HCoV) OC43. In a continuing effort to decipher the molecular mechanism underlying IFITM differential modulation of HCoV entry, we investigated the roles of structural motifs important for IFITM protein posttranslational modifications, intracellular trafficking, and oligomerization in modulating the entry of five HCoVs. We found that three distinct mutations in IFITM1 or IFITM3 converted the host restriction factors to enhance entry driven by the spike proteins of severe acute respiratory syndrome coronavirus (SARS-CoV) and/or Middle East respiratory syndrome coronavirus (MERS-CoV). First, replacement of IFITM3 tyrosine 20 with either alanine or aspartic acid to mimic unphosphorylated or phosphorylated IFITM3 reduced its activity to inhibit the entry of HCoV-NL63 and -229E but enhanced the entry of SARS-CoV and MERS-CoV. Second, replacement of IFITM3 tyrosine 99 with either alanine or aspartic acid reduced its activity to inhibit the entry of HCoV-NL63 and SARS-CoV but promoted the entry of MERS-CoV. Third, deletion of the carboxyl-terminal 12 amino acid residues from IFITM1 enhanced the entry of MERS-CoV and HCoV-OC43. These findings suggest that these residues and structural motifs of IFITM proteins are key determinants for modulating the entry of HCoVs, most likely through interaction with viral and/or host cellular components at the site of viral entry to modulate the fusion of viral envelope and cellular membranes. IMPORTANCE The differential effects of IFITM proteins on the entry of HCoVs that utilize divergent entry pathways and membrane fusion mechanisms even when using the same receptor make the HCoVs a valuable system for comparative investigation of the molecular mechanisms

  2. The determination of pesticide residues in local vegetables by means of neutron activation technique

    International Nuclear Information System (INIS)

    Mongkolphantha, S.; Karasuddhi, P.; Yamkate, P.; Serichareonsatit, N.

    1975-01-01

    Analytical methods based on neutron activation have been developed for studying pesticides residues of bromine, arsenic and mercury in local vegetables and fruits. The concentration of bromine, arsenic and mercury in samples are enriched prior to neutron irradiations by a technique of dry-ashing and freeze-drying for the determination of arsenic, bromine and mercury respectively. The element bromine is determined instrumentally while arsenic and mercury are determined destructively using a distillation technique. The limit of detection under the conditions used for bromine, arsenic and mercury as obtained are 0.01, 0.001 and 0.0001 microgram respectively. A total of 45 varieties of vegetables and 20 varieties of fruits are analyzed. The results of the investigation and the concentration range in part per million of bromine, arsenic and mercury are also presented

  3. Removal of dyes from aqueous solutions using activated carbon prepared from rice husk residue.

    Science.gov (United States)

    Li, Yaxin; Zhang, Xian; Yang, Ruiguang; Li, Guiying; Hu, Changwei

    2016-01-01

    The treatment of dye wastewater by activated carbon (AC) prepared from rice husk residue wastes was studied. Batch adsorption studies were conducted to investigate the effects of contact time, initial concentration (50-450 mg/L), pH (3-11) and temperature (30-70 °C) on the removal of methylene blue (MB), neutral red, and methyl orange. Kinetic investigation revealed that the adsorption of dyes followed pseudo-second-order kinetics. The results suggested that AC was effective to remove dyes, especially MB, from aqueous solutions. Desorption studies found that chemisorption by the adsorbent might be the major mode of dye removal. Fourier transform infrared results suggested that dye molecules were likely to combine with the O-H and P=OOH groups of AC.

  4. Enhancement of methylbenzene adsorption capacity through cetyl trimethyl ammonium bromide-modified activated carbon derived from Astragalus residue

    Science.gov (United States)

    Feng, Ningchuan; Zhang, Yumei; Fan, Wei; Zhu, Meilin

    2018-02-01

    Activated carbon was prepared from astragalus residue by KOH and then treated with cetyl trimethyl ammonium bromide (CTAB) and used for the removal of methylbenzene from aqueous solution. The samples were characterized by FTIR, XRD, SEM and Boehm titration. The results showed that CTAB changed the physicochemical properties of activated carbon significantly. The isotherm adsorption studies of methylbenzene onto the astragalus residue activated carbon (ASC) and CTAB-modified astragalus residue activated carbon (ASCCTAB) were examined by using batch techniques and agreed well with the Langmuir model. The maximum adsorption capacity of ASC and ASC-CTAB for methylbenzene determined from the Langmuir model was183.56 mg/g and 235.18 mg/g, respectively. The results indicated that using CTAB as a modifier for ASC modification could markedly enhance the methylbenzene removal from water.

  5. C-terminal tyrosine residues modulate the fusion activity of the Hendra virus fusion protein.

    Science.gov (United States)

    Popa, Andreea; Pager, Cara Teresia; Dutch, Rebecca Ellis

    2011-02-15

    The paramyxovirus family includes important human pathogens such as measles, mumps, respiratory syncytial virus, and the recently emerged, highly pathogenic Hendra and Nipah viruses. The viral fusion (F) protein plays critical roles in infection, promoting both the virus-cell membrane fusion events needed for viral entry as well as cell-cell fusion events leading to syncytia formation. We describe the surprising finding that addition of the short epitope HA tag to the cytoplasmic tail (CT) of the Hendra virus F protein leads to a significant increase in the extent of cell-cell membrane fusion. This increase was not due to alterations in surface expression, cleavage state, or association with lipid microdomains. Addition of a Myc tag of similar length did not alter Hendra F protein fusion activity, indicating that the observed stimulation was not solely a result of lengthening the CT. Three tyrosine residues within the HA tag were critical for the increase in the extent of fusion, suggesting C-terminal tyrosines may modulate Hendra fusion activity. The effects of addition of the HA tag varied with other fusion proteins, as parainfluenza virus 5 F-HA showed a decreased level of surface expression and no stimulation of fusion. These results indicate that additions to the C-terminal end of the F protein CT can modulate protein function in a sequence specific manner, reinforcing the need for careful analysis of epitope-tagged glycoproteins. In addition, our results implicate C-terminal tyrosine residues in the modulation of the membrane fusion reaction promoted by these viral glycoproteins.

  6. Preparation and Performance Evaluation of Biochars from Neem Seed Active Substance Extracted Residues (NSASER)

    Science.gov (United States)

    Li, Jialei; Lu, Shuwen

    2017-05-01

    Neem seed active substance extracted residues (NSASER) is industrial by-product, which is often discarded as a waste. It would lead to a certain degree of harm to the environment. The aim of this study was to prepare the biochars with neem seed active substance extracted residues (NSASER) under the anaerobic pyrolysis conditions. The pyrolysis process was studied with different pyrolysis power (200W, 500W, 800W, 1100W and 1400W), and the performance of the prepared biochars was evaluated. The results showed that the required time was to complete the pyrolysis process that gradually decreased with pyrolysis power increased from 200 to 1400 W, and the final pyrolysis temperature was to complete the pyrolysis process that increased with pyrolysis power increased from 200 to 1400 W. The biochars yield decreased with pyrolysis power increased from 200 to 1400 W, and the biochars yield has the maximum value when the pyrolysis power was of 200W. And the prepared biochars still had some characteristics of the plant cell and kept uniform porous structure, which was beneficial to absorb the small molecule substance. The water content of the prepared biochars was 7.18±0.53, the ash content of the prepared biochars was 5.92±0.31 and the fixed carbon content of the prepared biochars was 81.27±0.89. Compared with the bamboo charcoal, the performance index of the prepared biochars was in according with National Standard of the People’s Republic of China GBT26913-2011 of the bamboo charcoal. The prepared biochars had a potential value in application.

  7. In vitro residual activity of phenylalanine hydroxylase variants and correlation with metabolic phenotypes in PKU.

    Science.gov (United States)

    Trunzo, Roberta; Santacroce, Rosa; Shen, Nan; Jung-Klawitter, Sabine; Leccese, Angelica; De Girolamo, Giuseppe; Margaglione, Maurizio; Blau, Nenad

    2016-12-05

    Hyperphenylalaninemias (HPAs) are genetic diseases predominantly caused by a wide range of variants in the phenylalanine hydroxylase (PAH) gene. In vitro expression analysis of PAH variants offers the opportunity to elucidate the molecular mechanisms involved in HPAs and to clarify whether a disease-associated variant is genuinely pathogenic, while investigating the severity of a metabolic phenotype, and determining how a variant exerts its deleterious effects on the PAH enzyme. To study the effects of gene variants on PAH activity, we investigated eight variants: c.611A>G (p.Y204C), c.635T>C (p.L212P), c.746T>C (p.L249P), c.745C>T (p.L249F), c.809G>A (p.R270K), c.782G>C (p.R261P), c.587C>A (p.S196Y) and c.1139C>T (p.T380M), associated with different phenotypic groups. Transient expression of mutant full-length cDNAs in COS-7 cells yielded PAH proteins with PAH activity levels between 7% and 51% compared to the wild-type enzyme. With one exception (p.Y204C, which had no significant impact on PAH function), lower PAH activity was associated with a more severe phenotype (e.g. p.L249P with 7% PAH activity, 100% of classic PKU and no BH 4 responsiveness), while higher activity correlated with milder phenotypes (e.g. p.T380M with 28% PAH activity, 97% of mild HPA and 83% of BH 4 responsiveness). The results of the in vitro residual PAH activity have major implications, both for our understanding of genotype-phenotype correlations, and thereby existing inconsistencies, but also for the elucidation of the molecular basis of tetrahydrobiopterin (BH 4 ) responsiveness. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Mild thyroid peroxidase deficiency caused by TPO mutations with residual activity: Correlation between clinical phenotypes and enzymatic activity.

    Science.gov (United States)

    Narumi, Satoshi; Fox, Larry A; Fukudome, Keisuke; Sakaguchi, Zenichi; Sugisawa, Chiho; Abe, Kiyomi; Kameyama, Kaori; Hasegawa, Tomonobu

    2017-11-29

    Thyroid peroxidase (TPO) deficiency, caused by biallelic TPO mutations, is a well-established genetic form of congenital hypothyroidism (CH). More than 100 patients have been published, and the patients have been diagnosed mostly in the frame of newborn screening (NBS) programs. Correlation between clinical phenotypes and TPO activity remains unclear. Here, we report clinical and molecular findings of two unrelated TPO mutation-carrying mildly hypothyroid patients. The two patients were born at term after an uneventful pregnancy and delivery, and were NBS negative. They sought medical attention due to goiter at age 8 years. Evaluation of the thyroid showed mild elevation of serum TSH levels, normal or slightly low serum T 4 levels, high serum T 3 to T 4 molar ratio, high serum thyroglobulin levels, and high thyroidal 123 I uptake. We performed next-generation sequencing-based genetic screening, and found that one patient was compound heterozygous for two novel TPO mutations (p.Asp224del; c.820-2A>G), and the other was homozygous for a previously known mutation (p.Trp527Cys). In vitro functional analyses using HEK293 cells showed that the two amino acid-altering mutations (p.Asp224del and p.Trp527Cys) caused partial loss of the enzymatic activity. In conclusion, we report that TPO mutations with residual activity are associated with mild TPO deficiency, which is clinically characterized by marked goiter, mild TSH elevation, high serum T 3 to T 4 molar ratio, and high serum thyroglobulin levels. Our findings illuminate the hitherto under-recognized correlation between clinical phenotypes and residual enzymatic activity among patients with TPO deficiency.

  9. Inhibition of allergic dermal inflammation by the novel imidazopyridazine derivative TAK-427 in a guinea pig experimental model of eczema.

    Science.gov (United States)

    Fukuda, Shigeru; Midoro, Katsuo; Kamei, Takayuki; Gyoten, Michiyo; Kawano, Yasuhiko; Ashida, Yasuko; Nagaya, Hideaki

    2002-12-01

    Antigen challenge by patch ovalbumin emulsion induced an eczema-like skin lesion in epicutaneously sensitized guinea pigs. Diseased skin sites were macroscopically characterized by manifestations of dermatitis, such as erythema, edema, and papules, and microscopically characterized by acanthosis, spongiosis, and dermal infiltration by eosinophils. Using such lesions as a model of eczema, we evaluated the potential value of TAK-427 [2-[6-[[3-[4-(diphenylmethoxy)piperidino]propyl]amino] imidazo[1,2-b]pyridazin-2-yl]-2-methylpropionic acid dihydrate] as a therapeutic agent for atopic dermatitis by comparing it with dexamethasone and antihistamines. TAK-427 (0.3-30 mg/kg, p.o.) and dexamethasone (3 and 10 mg/kg, p.o.) inhibited eosinophil infiltration into the skin and ameliorated the dermatitis manifestations and epidermal damage. By contrast, none of the antihistamines tested (azelastine, ketotifen, terfenadine, and cetirizine) suppressed the eosinophil infiltration or dermatitis manifestations. To elucidate the mechanism by which TAK-427 inhibited the development of eczema, we investigated cytokine expression in the affected skin. Both TAK-427 and dexamethasone suppressed the increased mRNA expression of interleukin (IL)-13, granulocyte-macrophage colony-stimulating factor, IL-1alpha, tumor necrosis factor-alpha, interferon-gamma, and IL-8, but not IL-10, suggesting that TAK-427 inhibits allergic inflammation of the skin leading to the development of eczema by inhibiting the expression of proinflammatory cytokines after antigen challenge.

  10. Kombinasi Regresi Tak Bias Ridge dengan Regresi Komponen Utama untuk Mengatasi Masalah Multikolinieritas

    Directory of Open Access Journals (Sweden)

    Fitriana Novitasari

    2017-07-01

    Full Text Available Metode kuadrat terkecil (MKT merupakan salah satu metode penaksir parameter regresi. Dalam metode MKT terdapat salah satu asumsi yang harus dipenuhi yaitu tidak adanya multikolinieritas. Multikolinieritas merupakan hubungan linier antara sesama variabel bebas ( yang akan berakibat pada ragam penduga koefisisen regresi menjadi besar sehingga menyebabkan selang kepercayaan untuk parameter regresi cenderung akan lebih lebar. Salah satu metode yang dapat digunakan untuk mengatasi multikolinieritas adalah regresi MCRR, dimana metode MCRR ini merupakan kombinasi antara regresi tak bias ridge (URR dan regresi komponen utama. Makalah ini akan membahas penanganan multikolinieritas dengan menggunakan MCRR, sebagai bahan aplikasi akan digunakan data tingkat produksi Crude Palm Oil (CPO.

  11. Production of hydrolysate from processed Nile tilapia (Oreochromis niloticus residues and assessment of its antioxidant activity

    Directory of Open Access Journals (Sweden)

    Daniela Miotto BERNARDI

    2016-01-01

    Full Text Available Abstract The objective of this work was to produce protein hydrolysates from by-products of the Nile tilapia fileting process, and to assess the effects of different hydrolysis times on the antioxidant activity of the hydrolysed animal-based protein, in free form and incorporated into a food matrix. Gutted tilapia heads and carcasses were hydrolysed by Alcalase® for different hydrolysis times producing six hydrolysates. The protein content, degree of hydrolysis, reverse-phase high-performance liquid chromatography, and antioxidant activity by the ORAC, FRAP and TEAC methods were analysed. Three mini-hamburger formulations were produced and the lipidic oxidation of mini-hamburger was determined by TBARS. The protein contained in the residue was completely recovered in the process. The hydrolysates varied in their degree of hydrolysis, but presented similar levels of antioxidant activity. In the mini-hamburgers the hydrolysate was capable of delaying oxidation after 7 days of storage. Hydrolysis of tilapia processing by-products produced peptides may be used in the formulation of functional foods.

  12. Antioxidant, Anti-Tyrosinase and Anti-Inflammatory Activities of Oil Production Residues from Camellia tenuifloria

    Science.gov (United States)

    Chiou, Shu-Yuan; Ha, Choi-Lan; Wu, Pei-Shan; Yeh, Chiu-Ling; Su, Ying-Shan; Li, Man-Po; Wu, Ming-Jiuan

    2015-01-01

    Camellia tenuifloria is an indigenous Camellia species used for the production of camellia oil in Taiwan. This study investigated for the first time the potential antioxidant, anti-tyrosinase and anti-inflammatory activities of oil production byproducts, specifically those of the fruit shell, seed shell, and seed pomace from C. tenuifloria. It was found that the crude ethanol extract of the seed shell had the strongest DPPH scavenging and mushroom tyrosinase inhibitory activities, followed by the fruit shell, while seed pomace was the weakest. The IC50 values of crude extracts and fractions on monophenolase were smaller than diphenolase. The phenolic-rich methanol fraction of seed shell (SM) reduced nitric oxide (NO) production, and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. It also repressed the expression of IL-1β, and secretion of prostaglandin E2 (PGE2) and IL-6 in response to LPS. SM strongly stimulated heme oxygenase 1 (HO-1) expression and addition of zinc protoporphyrin (ZnPP), a HO-1 competitive inhibitor, reversed the inhibition of NO production, indicating the involvement of HO-1 in its anti-inflammatory activity. The effects observed in this study provide evidence for the reuse of residues from C. tenuifloria in the food additive, medicine and cosmetic industries. PMID:26690417

  13. Antioxidant, Anti-Tyrosinase and Anti-Inflammatory Activities of Oil Production Residues from Camellia tenuifloria

    Directory of Open Access Journals (Sweden)

    Shu-Yuan Chiou

    2015-12-01

    Full Text Available Camellia tenuifloria is an indigenous Camellia species used for the production of camellia oil in Taiwan. This study investigated for the first time the potential antioxidant, anti-tyrosinase and anti-inflammatory activities of oil production byproducts, specifically those of the fruit shell, seed shell, and seed pomace from C. tenuifloria. It was found that the crude ethanol extract of the seed shell had the strongest DPPH scavenging and mushroom tyrosinase inhibitory activities, followed by the fruit shell, while seed pomace was the weakest. The IC50 values of crude extracts and fractions on monophenolase were smaller than diphenolase. The phenolic-rich methanol fraction of seed shell (SM reduced nitric oxide (NO production, and inducible nitric oxide synthase (iNOS expression in lipopolysaccharide (LPS-stimulated RAW 264.7 cells. It also repressed the expression of IL-1β, and secretion of prostaglandin E2 (PGE2 and IL-6 in response to LPS. SM strongly stimulated heme oxygenase 1 (HO-1 expression and addition of zinc protoporphyrin (ZnPP, a HO-1 competitive inhibitor, reversed the inhibition of NO production, indicating the involvement of HO-1 in its anti-inflammatory activity. The effects observed in this study provide evidence for the reuse of residues from C. tenuifloria in the food additive, medicine and cosmetic industries.

  14. Residues in the alternative reading frame tumor suppressor that influence its stability and p53-independent activities

    Energy Technology Data Exchange (ETDEWEB)

    Tommaso, Anne di [Pole Biologie Sante, UMR 6187 CNRS, Pathologies Moleculaire de l' Adressage et de la Signalisation, Universite de Poitiers, Poitiers (France); Hagen, Jussara; Tompkins, Van [Department of Pharmacology, The University of Iowa, College of Medicine, Iowa City, IA (United States); Muniz, Viviane [Molecular and Cellular Biology Program, The University of Iowa, College of Medicine, Iowa City, IA (United States); Dudakovic, Amel [Department of Pharmacology, The University of Iowa, College of Medicine, Iowa City, IA (United States); Kitzis, Alain [Pole Biologie Sante, UMR 6187 CNRS, Pathologies Moleculaire de l' Adressage et de la Signalisation, Universite de Poitiers, Poitiers (France); CHU de Poitiers, Poitiers (France); Ladeveze, Veronique [Pole Biologie Sante, UMR 6187 CNRS, Pathologies Moleculaire de l' Adressage et de la Signalisation, Universite de Poitiers, Poitiers (France); Quelle, Dawn E., E-mail: dawn-quelle@uiowa.edu [Department of Pharmacology, The University of Iowa, College of Medicine, Iowa City, IA (United States); Molecular and Cellular Biology Program, The University of Iowa, College of Medicine, Iowa City, IA (United States)

    2009-04-15

    The Alternative Reading Frame (ARF) protein suppresses tumorigenesis through p53-dependent and p53-independent pathways. Most of ARF's anti-proliferative activity is conferred by sequences in its first exon. Previous work showed specific amino acid changes occurred in that region during primate evolution, so we programmed those changes into human p14ARF to assay their functional impact. Two human p14ARF residues (Ala{sup 14} and Thr{sup 31}) were found to destabilize the protein while two others (Val{sup 24} and Ala{sup 41}) promoted more efficient p53 stabilization and activation. Despite those effects, all modified p14ARF forms displayed robust p53-dependent anti-proliferative activity demonstrating there are no significant biological differences in p53-mediated growth suppression associated with simian versus human p14ARF residues. In contrast, p53-independent p14ARF function was considerably altered by several residue changes. Val{sup 24} was required for p53-independent growth suppression whereas multiple residues (Val{sup 24}, Thr{sup 31}, Ala{sup 41} and His{sup 60}) enabled p14ARF to block or reverse the inherent chromosomal instability of p53-null MEFs. Together, these data pinpoint specific residues outside of established p14ARF functional domains that influence its expression and signaling activities. Most intriguingly, this work reveals a novel and direct role for p14ARF in the p53-independent maintenance of genomic stability.

  15. Residues in the alternative reading frame tumor suppressor that influence its stability and p53-independent activities

    International Nuclear Information System (INIS)

    Tommaso, Anne di; Hagen, Jussara; Tompkins, Van; Muniz, Viviane; Dudakovic, Amel; Kitzis, Alain; Ladeveze, Veronique; Quelle, Dawn E.

    2009-01-01

    The Alternative Reading Frame (ARF) protein suppresses tumorigenesis through p53-dependent and p53-independent pathways. Most of ARF's anti-proliferative activity is conferred by sequences in its first exon. Previous work showed specific amino acid changes occurred in that region during primate evolution, so we programmed those changes into human p14ARF to assay their functional impact. Two human p14ARF residues (Ala 14 and Thr 31 ) were found to destabilize the protein while two others (Val 24 and Ala 41 ) promoted more efficient p53 stabilization and activation. Despite those effects, all modified p14ARF forms displayed robust p53-dependent anti-proliferative activity demonstrating there are no significant biological differences in p53-mediated growth suppression associated with simian versus human p14ARF residues. In contrast, p53-independent p14ARF function was considerably altered by several residue changes. Val 24 was required for p53-independent growth suppression whereas multiple residues (Val 24 , Thr 31 , Ala 41 and His 60 ) enabled p14ARF to block or reverse the inherent chromosomal instability of p53-null MEFs. Together, these data pinpoint specific residues outside of established p14ARF functional domains that influence its expression and signaling activities. Most intriguingly, this work reveals a novel and direct role for p14ARF in the p53-independent maintenance of genomic stability.

  16. Therapeutic effect of methyl salicylate 2-O-β-d-lactoside on LPS-induced acute lung injury by inhibiting TAK1/NF-kappaB phosphorylation and NLRP3 expression.

    Science.gov (United States)

    Yang, Shengqian; Yu, Ziru; Yuan, Tianyi; Wang, Lin; Wang, Xue; Yang, Haiguang; Sun, Lan; Wang, Yuehua; Du, Guanhua

    2016-11-01

    Acute lung injury (ALI), characterized by pulmonary edema and inflammatory cell infiltration, is a common syndrome of acute hypoxemic respiratory failure. Methyl salicylate 2-O-β-d-lactoside (MSL), a natural derivative of salicylate extracted from Gaultheria yunnanensis (Franch.) Rehder, was reported to have potent anti-inflammatory effects on the progression of collagen or adjuvant-induced arthritis in vivo and in vitro. The aim of this study is to investigate the therapeutic effect of MSL on lipopolysaccharide (LPS)-induced acute lung injury and reveal underlying molecular mechanisms. Our results showed that MSL significantly ameliorated pulmonary edema and histological severities, and inhibited IL-6 and IL-1β production in LPS-induced ALI mice. MSL also reduced MPO activity in lung tissues and the number of inflammatory cells in BALF. Moreover, we found that MSL significantly inhibited LPS-induced TAK1 and NF-κB p65 phosphorylation, as well as the expression of NLRP3 protein in lung tissues. Furthermore, MSL significantly inhibited LPS-induced TAK1 and NF-κB p65 phosphorylation in Raw264.7 cells. In addition, MSL significantly inhibited nuclear translocation of NF-κB p65 in cells treated with LPS in vitro. Taken together, our results suggested that MSL exhibited a therapeutic effect on LPS-induced ALI by inhibiting TAK1/NF-κB phosphorylation and NLRP3 expression. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Elimination of textile dyes using activated carbons prepared from vegetable residues and their characterization.

    Science.gov (United States)

    Peláez-Cid, Alejandra-Alicia; Herrera-González, Ana-María; Salazar-Villanueva, Martín; Bautista-Hernández, Alejandro

    2016-10-01

    In this study, three mesoporous activated carbons prepared from vegetable residues were used to remove acid, basic, and direct dyes from aqueous solutions, and reactive and vat dyes from textile wastewater. Granular carbons obtained by chemical activation at 673 K with phosphoric acid from prickly pear peels (CarTunaQ), broccoli stems (CarBrocQ), and white sapote seeds (CarZapQ) were highly efficient for the removal of dyes. Adsorption equilibrium studies were carried out in batch systems and treated with Langmuir and Freundlich isotherms. The maximum adsorption capacities calculated from the Langmuir isotherms ranged between 131.6 and 312.5 mg/g for acid dyes, and between 277.8 and 500.0 mg/g for basic dyes at 303 K. Our objective in this paper was to show that vegetable wastes can serve as precursors for activated carbons that can be used for the adsorption of dyes. Specifically CarBrocQ was the best carbon produced for the removal of textile dyes. The color removal of dyes present in textile wastewaters was compared with that of a commercial powdered carbon, and it was found that the carbons produced using waste material reached similar efficiency levels. Carbon samples were characterized by bulk density, point of zero charge, thermogravimetric analysis, elemental analysis, Fourier transform infrared spectroscopy, scanning electron microscopy, methylene blue adsorption isotherms at 303 K, and nitrogen adsorption isotherms at 77 K (SBET). The results show that the activated carbons possess a large specific surface area (1025-1177 m(2)/g) and high total pore volume (1.06-2.16 cm(3)/g) with average pore size diameters between 4.1 and 8.4 nm. Desorption and regeneration tests were made to test the viability of reusing the activated carbons. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Atividade residual de diuron, oxyfluorfen e prometryne no controle de Euphorbia heterophylla Residual activity of diuron, oxyfluorfen, and prometryne for Euphorbia heterophylla control

    Directory of Open Access Journals (Sweden)

    R.S. Oliveira Jr.

    2012-09-01

    Full Text Available As aplicações de herbicidas em pré-emergência têm por finalidade a obtenção da atividade residual no início do ciclo das culturas. Assim, o objetivo deste trabalho foi avaliar a atividade residual dos herbicidas diuron, oxyfluorfen e prometryne, aplicados isoladamente ou em misturas, no controle de Euphorbia heterophylla. Oito experimentos foram conduzidos em casa de vegetação, aplicando-se doses dos herbicidas ou das misturas aos 30, 20, 10 e 0 dias antes da semeadura da planta daninha (DAS. Com o diuron e prometryne, foram observados controles satisfatórios até 20 DAS nas doses a partir de 1,07 e 1,6 kg ha-1, respectivamente. Quanto ao oxyfluorfen, foi registrado um período residual inferior, obtendo-se controle mínimo de 80% até 10 DAS nas doses a partir de 0,324 kg ha-1. Em relação às misturas dos herbicidas, a mistura diuron+prometryne promoveu controle superior a 85% por períodos de até 30 dias, quando aplicada na menor dose (1+2 kg ha-1, e de 20 dias, quando aplicada na dose de 2+1 kg ha-1. Visando obter esse mesmo patamar de controle por 30 dias, foi necessário 1+0,288 kg ha-1 da mistura diuron+oxyfluorfen. A mistura prometryne+oxyfluorfen apresentou um mínimo de 80% de controle no período de 10 dias, quando utilizada a dose de 1+0,192 kg ha-1.Pre-emergence herbicide applications are designed to obtain residual activity at the beginning of the crop cycle. The objective of this study was to evaluate the residual activity of diuron, oxyfluorfen, and prometryne, applied alone or in mixture, to control Euphorbia heterophylla. Eight experiments were conducted under greenhouse conditions, by applying herbicide doses or mixtures at 30, 20, 10, and 0 days before weed sowing (DBWS.With diuron and prometryne, satisfactory controls were observed at doses up to 20 DBWS, from 1.07 to 1.6 kg ha-1, respectively. As for oxyfluorfen, a lower residual period was verified, with a minimum gaining control of at least 80% being obtained

  19. Modeled Oceanic Response and Sea Surface Cooling to Typhoon Kai-Tak

    Directory of Open Access Journals (Sweden)

    Yu-Heng Tseng

    2010-01-01

    Full Text Available An ocean response to typhoon Kai-Tak is simulated using an accurate fourth-order, basin-scale ocean model. The surface winds of typhoon Kai-Tak were obtained from QuikSCAT satellite images blended with the ECMWF wind fields. An intense nonlinear mesoscale eddy is generated in the northeast South China Sea (SCS with a Rossby number of O(1 and on a 50 - 100 km horizontal scale. Inertial oscillation is clearly observed. Advection dominates as a strong wind shear drives the mixed layer flows outward, away from the typhoon center, thus forcing upwelling from deep levels with a high upwelling velocity (> 30 m day-1. A drop in sea surface temperature (SST of more than 9°C is found in both observation and simulation. We attribute this significant SST drop to the influence of the slow moving typhoon, initial stratification and bathymetry-induced upwelling in the northeast of the SCS where the typhoon hovered.

  20. Social Marketing in Malaysia: Cognitive, Affective, and Normative Mediators of the TAK NAK Antismoking Advertising Campaign.

    Science.gov (United States)

    Lee, Wonkyong Beth; Fong, Geoffrey T; Dewhirst, Timothy; Kennedy, Ryan D; Yong, Hua-Hie; Borland, Ron; Awang, Rahmat; Omar, Maizurah

    2015-01-01

    Antismoking mass media campaigns are known to be effective as part of comprehensive tobacco control programs in high-income countries, but such campaigns are relatively new in low- and middle-income countries and there is a need for strong evaluation studies from these regions. This study examines Malaysia's first national antismoking campaign, TAK NAK. The data are from the International Tobacco Control Malaysia Survey, which is an ongoing cohort survey of a nationally representative sample of adult smokers (18 years and older; N = 2,006). The outcome variable was quit intentions of adult smokers, and the authors assessed the extent to which quit intentions may have been strengthened by exposure to the antismoking campaign. The authors also tested whether the impact of the campaign on quit intentions was related to cognitive mechanisms (increasing thoughts about the harm of smoking), affective mechanisms (increasing fear from the campaign), and perceived social norms (increasing perceived social disapproval about smoking). Mediational regression analyses revealed that thoughts about the harm of smoking, fear arousal, and social norms against smoking mediated the relation between TAK NAK impact and quit intentions. Effective campaigns should prompt smokers to engage in both cognitive and affective processes and encourage consideration of social norms about smoking in their society.

  1. Mutational and structural analyses of Caldanaerobius polysaccharolyticus Man5B reveal novel active site residues for family 5 glycoside hydrolases.

    Directory of Open Access Journals (Sweden)

    Takuji Oyama

    Full Text Available CpMan5B is a glycoside hydrolase (GH family 5 enzyme exhibiting both β-1,4-mannosidic and β-1,4-glucosidic cleavage activities. To provide insight into the amino acid residues that contribute to catalysis and substrate specificity, we solved the structure of CpMan5B at 1.6 Å resolution. The structure revealed several active site residues (Y12, N92 and R196 in CpMan5B that are not present in the active sites of other structurally resolved GH5 enzymes. Residue R196 in GH5 enzymes is thought to be strictly conserved as a histidine that participates in an electron relay network with the catalytic glutamates, but we show that an arginine fulfills a functionally equivalent role and is found at this position in every enzyme in subfamily GH5_36, which includes CpMan5B. Residue N92 is required for full enzymatic activity and forms a novel bridge over the active site that is absent in other family 5 structures. Our data also reveal a role of Y12 in establishing the substrate preference for CpMan5B. Using these molecular determinants as a probe allowed us to identify Man5D from Caldicellulosiruptor bescii as a mannanase with minor endo-glucanase activity.

  2. Mutational and structural analyses of Caldanaerobius polysaccharolyticus Man5B reveal novel active site residues for family 5 glycoside hydrolases.

    Science.gov (United States)

    Oyama, Takuji; Schmitz, George E; Dodd, Dylan; Han, Yejun; Burnett, Alanna; Nagasawa, Naoko; Mackie, Roderick I; Nakamura, Haruki; Morikawa, Kosuke; Cann, Isaac

    2013-01-01

    CpMan5B is a glycoside hydrolase (GH) family 5 enzyme exhibiting both β-1,4-mannosidic and β-1,4-glucosidic cleavage activities. To provide insight into the amino acid residues that contribute to catalysis and substrate specificity, we solved the structure of CpMan5B at 1.6 Å resolution. The structure revealed several active site residues (Y12, N92 and R196) in CpMan5B that are not present in the active sites of other structurally resolved GH5 enzymes. Residue R196 in GH5 enzymes is thought to be strictly conserved as a histidine that participates in an electron relay network with the catalytic glutamates, but we show that an arginine fulfills a functionally equivalent role and is found at this position in every enzyme in subfamily GH5_36, which includes CpMan5B. Residue N92 is required for full enzymatic activity and forms a novel bridge over the active site that is absent in other family 5 structures. Our data also reveal a role of Y12 in establishing the substrate preference for CpMan5B. Using these molecular determinants as a probe allowed us to identify Man5D from Caldicellulosiruptor bescii as a mannanase with minor endo-glucanase activity.

  3. Toxicity and residual activity of methoxyfenozide and tebufenozide to codling moth (Lepidoptera: Tortricidae) and oriental fruit moth (Lepidoptera: Tortricidae).

    Science.gov (United States)

    Borchert, Daniel M; Walgenbach, James F; Kennedy, George G; Long, John W

    2004-08-01

    A series of studies were conducted to examine the residual activity and toxicity of the ecdysone agonists tebufenozide and methoxyfenozide to codling moth, Cydia pomonella (L.), and oriental fruit moth, Grapholita molesta (Busck), in North Carolina apple systems. Methoxyfenozide exhibited greater activity than tebufenozide against codling moth eggs in dose-response bioassays, with a 4.5- and 5.3-fold lower LC50 value to eggs laid on fruit treated before or after oviposition, respectively. Oriental fruit moth eggs were 57- and 12-fold less sensitive to methoxyfenozide than were codling moth eggs on fruit treated before and after oviposition, respectively. Methoxyfenozide was effective in reducing larval entries of both codling moth and oriental fruit moth in field residual activity bioassays, exhibiting activity for at least 28 d after application. Residue breakdown on fruit was approximately 80% at 28 d after treatment for both methoxyfenozide and tebufenozide, with the most rapid residue decline (60%) occurring during the first 14 d after application. Two applications of methoxyfenozide applied at 14-d intervals provided better canopy coverage and higher residue levels than one application. Spray volume (683 versus 2,057 liters/ha) did not affect the efficacy of methoxyfenozide. Leaf and fruit expansion during the season was measured to determine potential plant-growth dilution effects on residual activity. There was very little increase in leaf area after mid May, but increase in fruit surface area over the season was described by a second order polynomial regression. Implications for codling moth and oriental fruit moth management programs are discussed.

  4. Canine Distemper Virus Fusion Activation: Critical Role of Residue E123 of CD150/SLAM.

    Science.gov (United States)

    Khosravi, Mojtaba; Bringolf, Fanny; Röthlisberger, Silvan; Bieringer, Maria; Schneider-Schaulies, Jürgen; Zurbriggen, Andreas; Origgi, Francesco; Plattet, Philippe

    2016-02-01

    Measles virus (MeV) and canine distemper virus (CDV) possess tetrameric attachment proteins (H) and trimeric fusion proteins, which cooperate with either SLAM or nectin 4 receptors to trigger membrane fusion for cell entry. While the MeV H-SLAM cocrystal structure revealed the binding interface, two distinct oligomeric H assemblies were also determined. In one of the conformations, two SLAM units were sandwiched between two discrete H head domains, thus spotlighting two binding interfaces ("front" and "back"). Here, we investigated the functional relevance of both interfaces in activating the CDV membrane fusion machinery. While alanine-scanning mutagenesis identified five critical regulatory residues in the front H-binding site of SLAM, the replacement of a conserved glutamate residue (E at position 123, replaced with A [E123A]) led to the most pronounced impact on fusion promotion. Intriguingly, while determination of the interaction of H with the receptor using soluble constructs revealed reduced binding for the identified SLAM mutants, no effect was recorded when physical interaction was investigated with the full-length counterparts of both molecules. Conversely, although mutagenesis of three strategically selected residues within the back H-binding site of SLAM did not substantially affect fusion triggering, nevertheless, the mutants weakened the H-SLAM interaction recorded with the membrane-anchored protein constructs. Collectively, our findings support a mode of binding between the attachment protein and the V domain of SLAM that is common to all morbilliviruses and suggest a major role of the SLAM residue E123, located at the front H-binding site, in triggering the fusion machinery. However, our data additionally support the hypothesis that other microdomain(s) of both glycoproteins (including the back H-binding site) might be required to achieve fully productive H-SLAM interactions. A complete understanding of the measles virus and canine distemper virus

  5. The Phosphodiesterase 10A Selective Inhibitor TAK-063 Improves Cognitive Functions Associated with Schizophrenia in Rodent Models.

    Science.gov (United States)

    Shiraishi, Eri; Suzuki, Kazunori; Harada, Akina; Suzuki, Noriko; Kimura, Haruhide

    2016-03-01

    Cognitive deficits in various domains, including recognition memory, attention, impulsivity, working memory, and executive function, substantially affect functional outcomes in patients with schizophrenia. TAK-063 [1-[2-fluoro-4-(1H-pyrazol-1-yl)phenyl]-5-methoxy-3-(1-phenyl-1H-pyrazol-5-yl)pyridazin-4(1H)-one] is a potent and selective phosphodiesterase 10A inhibitor that produces antipsychotic-like effects in rodent models of schizophrenia. We evaluated the effects of TAK-063 on multiple cognitive functions associated with schizophrenia using naïve and drug-perturbed rodents. TAK-063 at 0.1 and 0.3 mg/kg p.o. improved time-dependent memory decay in object recognition in naïve rats. TAK-063 at 0.1 and 0.3 mg/kg p.o. increased accuracy rate, and TAK-063 at 0.3 mg/kg p.o. reduced impulsivity in a five-choice serial reaction time task in naïve rats. N-methyl-d-aspartate receptor antagonists, such as phencyclidine and MK-801 [(5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine], were used to induce working memory deficits relevant to schizophrenia in animals. TAK-063 at 0.3 mg/kg p.o. attenuated both phencyclidine-induced working memory deficits in a Y-maze test in mice and MK-801-induced working memory deficits in an eight-arm radial maze task in rats. An attentional set-shifting task using subchronic phencyclidine-treated rats was used to assess the executive function. TAK-063 at 0.3 mg/kg p.o. reversed cognitive deficits in extradimensional shifts. These findings suggest that TAK-063 has a potential to ameliorate deficits in multiple cognitive domains impaired in schizophrenia. Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.

  6. Enzyme active site mimics based on TriAzaCyclophane (TAC)-scaffolded peptides and amino acid residues

    NARCIS (Netherlands)

    Albada, H.B.

    2009-01-01

    This thesis describes the scope and limitations of the application of TriAzaCyclophane (TAC)-scaffolded peptides or amino acid residues as enzyme active site mimics, as ligands in asymmetric catalysis and as hydrolysis catalysts attached to vancomycin. For the mimicry of functional group enzymes, of

  7. A simple measurement of the Ni-63 diffusion profiles by residual activity method using a liquid scintillation counter

    Czech Academy of Sciences Publication Activity Database

    Rothová, Věra

    2013-01-01

    Roč. 729, NOV (2013), s. 702-706 ISSN 0168-9002 Institutional support: RVO:68081723 Keywords : Nickel-63 * Low-energy β-emitter * Solid scintillation counting * Diffusion * Gruzin′s residual activity method Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 1.316, year: 2013

  8. Active disease and residual damage in treated Wegener's granulomatosis: an observational study using pulmonary high-resolution computed tomography

    International Nuclear Information System (INIS)

    Komocsi, Andras; Reuter, Michael; Heller, Martin; Murakoezi, Henriette; Gross, Wolfgang L.; Schnabel, Armin

    2003-01-01

    The purpose of this study was to determine to what extent high-resolution computed tomography (HRCT) of the lungs can distinguish active inflammatory disease from inactive cicatricial disease in patients treated for Wegener's granulomatosis (WG). Twenty-eight WG patients with active pulmonary disease underwent a first HRCT examination immediately before standard immunosuppressive treatment and a second examination after clinical remission had been achieved. Lesions remaining after treatment were categorized as residual damage and were compared with findings during active disease to see by what features active and cicatricial disease can be distinguished. During active disease 17 patients had nodules/masses, 12 had ground-glass opacities, 6 had septal lines and 6 had non-septal lines. After treatment, ground-glass opacities had resolved completely. Nodules/masses had resolved in 8 patients and had diminished in 7 patients. Residual nodules were distinguished from nodules/masses in active disease by lack of cavitation and a diameter of mostly <15 mm. In one-third of patients lines resolved, but in 8 instances new lines evolved during immunosuppression. During a follow-up period of a median 26.5 months (range 20.0-33.8), patients with residual nodules or lines had no more relapses than patients with completely cleared lungs. Treated pulmonary WG leaves substantial residual damage. High-resolution CT does assist in the distinction between active and inactive lesions. Ground-glass opacities, cavitating nodules/masses and masses measuring more than 3 cm represent active disease ordinarily. Non-cavitary small nodules and septal or non-septal lines can be either active or cicatricial lesions. The nature of these lesions needs to be clarified by longitudinal observation. (orig.)

  9. Identification of critical residues for the activity and thermostability of Streptomyces sp. SK glucose isomerase.

    Science.gov (United States)

    Ben Hlima, Hajer; Bejar, Samir; Riguet, Jonas; Haser, Richard; Aghajari, Nushin

    2013-11-01

    The role of residue 219 in the physicochemical properties of D-glucose isomerase from Streptomyces sp. SK strain (SKGI) was investigated by site-directed mutagenesis and structural studies. Mutants G219A, G219N, and G219F were generated and characterized. Comparative studies of their physicochemical properties with those of the wild-type enzyme highlighted that mutant G219A displayed increased specific activity and thermal stability compared to that of the wild-type enzyme, while for G219N and G219F, these properties were considerably decreased. A double mutant, SKGI F53L/G219A, displayed a higher optimal temperature and a higher catalytic efficiency than both the G219A mutant and the wild-type enzyme and showed a half-life time of about 150 min at 85 °C as compared to 50 min for wild-type SKGI. Crystal structures of SKGI wild-type and G219A enzymes were solved to 1.73 and 2.15 Å, respectively, and showed that the polypeptide chain folds into two structural domains. The larger domain consists of a (β/α)8 unit, and the smaller domain forms a loop of α helices. Detailed analyses of the three-dimensional structures highlighted minor but important changes in the active site region as compared to that of the wild-type enzyme leading to a displacement of both metal ions, and in particular that in site M2. The structural analyses moreover revealed how the substitution of G219 by an alanine plays a crucial role in improving the thermostability of the mutant enzyme.

  10. Inhibition of TAK1 and/or JAK can rescue impaired chondrogenic differentiation of human mesenchymal stem cells in osteoarthritis-like conditions.

    Science.gov (United States)

    van Beuningen, Henk M; de Vries-van Melle, Marloes L; Vitters, Elly L; Schreurs, Wim; van den Berg, Wim B; van Osch, Gerjo J V M; van der Kraan, Peter M

    2014-08-01

    To rescue chondrogenic differentiation of human mesenchymal stem cells (hMSCs) in osteoarthritic conditions by inhibition of protein kinases. hMSCs were cultured in pellets. During early chondrogenic differentiation, these were exposed to osteoarthritic synovium-conditioned medium (OAS-CM), combined with the Janus kinase (JAK)-inhibitor tofacitinib and/or the transforming growth factor β-activated kinase 1 (TAK1)-inhibitor oxozeaenol. To evaluate effects on chondrogenesis, the glycosaminoglycan (GAG) content of the pellets was measured at the time that chondrogenesis was manifest in control cultures. Moreover, mRNA levels of matrix molecules and enzymes were measured during this process, using real-time polymerase chain reaction (RT-PCR). Initial experiments were performed with hMSCs from a fetal donor, and results of these studies were confirmed with hMSCs from adult donors. Exposure to OAS-CM resulted in pellets with a much lower GAG content, reflecting inhibited chondrogenic differentiation. This was accompanied by decreased mRNA levels of aggrecan, type II collagen, and Sox9, and increased levels of matrix metalloproteinase (MMP)1, MMP3, MMP13, ADAMTS4, and ADAMTS5. Both tofacitinib (JAK-inhibitor) and oxozeaenol (TAK1 inhibitor) significantly increased the GAG content of the pellets in osteoarthritis (OA)-like conditions. The combination of both protein kinase inhibitors showed an additive effect on GAG content. In agreement with this, in the presence of OAS-CM, both tofacitinib and oxozeaenol increased mRNA expression of sox9. The expression of aggrecan and type II collagen was also up-regulated, but this only reached significance for aggrecan after TAK1 inhibition. Both inhibitors decreased the mRNA levels of MMP1, 3, and 13 in the presence of OAS-CM. Moreover, oxozeaenol also significantly down-regulated the mRNA levels of aggrecanases ADAMTS4 and ADAMTS5. When combined, the inhibitors caused additive reduction of OA-induced MMP1 mRNA expression

  11. Analysis of Active-Site Amino-Acid Residues of Human Serum Paraoxonase Using Competitive Substrates

    National Research Council Canada - National Science Library

    Yeung, David T; Lenz, David E; Cerasoli, Douglas M

    2005-01-01

    ...(s) or its catalytic mechanism. Through site-directed mutagenesis studies, designed from a DFPase-like homology model, and from a crystal structure of a hybrid PONl molecule, amino-acid residues essential for enzyme function...

  12. Ne/přízvučné slovo tak ve dvou televizních talk show

    Czech Academy of Sciences Publication Activity Database

    Jílková, Lucie

    2017-01-01

    Roč. 54, 3/4 (2017), s. 4-20 ISSN 1212-5326 R&D Projects: GA ČR GA15-01116S Institutional support: RVO:68378092 Keywords : Czech word tak * DIALOG corpus * word stress * television talk show Subject RIV: AI - Linguistics OBOR OECD: Linguistics

  13. Antibacterial and residual antimicrobial activities against Enterococcus faecalis biofilm: A comparison between EDTA, chlorhexidine, cetrimide, MTAD and QMix

    Science.gov (United States)

    Zhang, Rui; Chen, Min; Lu, Yan; Guo, Xiangjun; Qiao, Feng; Wu, Ligeng

    2015-08-01

    We compared the antibacterial and residual antimicrobial activities of five root canal irrigants (17% EDTA,2% chlorhexidine,0.2% cetrimide, MTAD, and QMix) in a model of Enterococcus faecalis biofilm formation. Sixty dentin blocks with 3-week E. faecalis biofilm were divided into six equal groups and flushed with irrigant for 2 min or left untreated. A blank control group was also established. Antibacterial activities of the irrigants were evaluated by counting colony forming units. To test residual antimicrobial activities, 280 dentin blocks were divided into seven equal groups and flushed with irrigant for 2 min or left untreated and then incubated with E. faecalis suspension for 48 h, or used as a blank. No bacteria were observed in the blank control group. The number of viable E. faecalis was significantly fewer in the irrigant-treated groups compared with the untreated control (P < 0.05). Among the five irrigants, QMix had the strongest antibacterial activity. Residual antimicrobial activities of CHX were significantly higher at 12 h, 24 h and 36 h compared to untreated control (P < 0.05). All five root canal irrigants were effective to some extent against E. faecalis, but QMix and CHX had the strongest, and CHX the longest (up to 36 h), antimicrobial activity.

  14. Penentuan Distribusi Suhu pada Permukaan Geometri Tak Tentu Menggunakan Metode Random Walk

    Directory of Open Access Journals (Sweden)

    Balduyanus Yosep Godja

    2016-05-01

    Full Text Available Telah dilakukan penentuan distribusi suhu dalam keadaan tunak pada sebuah plat bergeometri tak tentu menggunakan metode Random Walk yang dilengkapi fungsi green. Setiap sisi plat dikondisikan bervariasi terhadap suhu dalam rentang 10°C sampai 100°C dengan 4 (empat konfigurasi berkeadaan steady. Persamaan Laplace yang mendeskripsikan permasalahan ini dihampiri dengan mensimulasikan sejumlah walker pada setiap titik domain permasalahan untuk kemudian secara acak disebar menuju ke setiap sisi plat. Hasil yang diperoleh untuk setiap kondisi plat menunjukkan kesalahan relatif terhadap solusi numerik metode iterasi jacobi yang telah menghampiri solusi analitik, secara rata-rata adalah 0,85%. Nilai kesalahan tersebut diperoleh dengan menggunakan 5000 walker. Penelitian ini juga mendapatkan bahwa akurasi hampiran ditentukan oleh banyaknya walker yang digunakan. Secara umum, semakin banyak jumlah walker yang digunakan maka akurasi hampiran akan semakin baik.

  15. Sustainable waste management by production of activated carbon from agroforestry residues

    Directory of Open Access Journals (Sweden)

    Victor Ntuli

    2013-01-01

    Full Text Available Agroforestry waste presents a problem for disposal and negatively impacts on the environment if left to rot or burn. The aim of this study was to reduce environmental problems associated with agroforestry waste by promoting the innovative use of such waste in the production of activated carbons (ACs using a low-cost production technique, and ultimately delivering more affordable water and effluent treatment adsorbents. Four varieties of ACs from four different agroforestry materials – pine (Pinus contorta cones (PC, Abies (Abies cilicica seeds (AS, maple (Acer ginnala seeds (MS and peach (Prunus persica stones (PS – were prepared by single-step steam pyrolysis and characterised. The raw materials were evaluated for AC yield while the respective ACs were evaluated on the basis of iodine number, phenol specific area, ash content, pH, moisture content and removal of metal ions, nitrates and sulphates from aqueous solution. The AC yields for PS, PC, AS and MS were found to be 23.0%, 18.0%, 17.8% and 14.6%, respectively. The yield for PS (23% is within the specified commercial limits of 20% to 40%. The phenol specific areas of the ACs ranged between 381 m2/g and 415 m2/g higher than the commercial lower limit (300 m2/g generally specified. The ACs also showed the capacity to remove heavy metal ions from their aqueous solutions. Removal of both nitrates and sulphates in raw water was greater than 50%. Although no quantitative analysis has been performed to date, it is envisaged that the production of AC from agroforestry wastes can contribute to the sustainable management of environmental pollution by these residues and the concomitant delivery of cheaper adsorbents.

  16. Acute and Chronic Effects of Dietary Lactose in Adult Rats Are not Explained by Residual Intestinal Lactase Activity

    Directory of Open Access Journals (Sweden)

    Bert J. M. van de Heijning

    2015-07-01

    Full Text Available Neonatal rats have a high intestinal lactase activity, which declines around weaning. Yet, the effects of lactose-containing products are often studied in adult animals. This report is on the residual, post-weaning lactase activity and on the short- and long-term effects of lactose exposure in adult rats. Acutely, the postprandial plasma response to increasing doses of lactose was studied, and chronically, the effects of a 30% lactose diet fed from postnatal (PN Day 15 onwards were evaluated. Intestinal lactase activity, as assessed both in vivo and in vitro, was compared between both test methods and diet groups (lactose vs. control. A 50%–75% decreased digestive capability towards lactose was observed from weaning into adulthood. Instillation of lactose in adult rats showed disproportionally low increases in plasma glucose levels and did not elicit an insulin response. However, gavages comprising maltodextrin gave rise to significant plasma glucose and insulin responses, indicative of a bias of the adult GI tract to digest glucose polymers. Despite the residual intestinal lactase activity shown, a 30% lactose diet was poorly digested by adult rats: the lactose diet rendered the animals less heavy and virtually devoid of body fat, whereas their cecum tripled in size, suggesting an increased bacterial fermentation. The observed acute and chronic effects of lactose exposure in adult rats cannot be explained by the residual intestinal lactase activity assessed.

  17. Acute and Chronic Effects of Dietary Lactose in Adult Rats Are not Explained by Residual Intestinal Lactase Activity.

    Science.gov (United States)

    van de Heijning, Bert J M; Kegler, Diane; Schipper, Lidewij; Voogd, Eline; Oosting, Annemarie; van der Beek, Eline M

    2015-07-08

    Neonatal rats have a high intestinal lactase activity, which declines around weaning. Yet, the effects of lactose-containing products are often studied in adult animals. This report is on the residual, post-weaning lactase activity and on the short- and long-term effects of lactose exposure in adult rats. Acutely, the postprandial plasma response to increasing doses of lactose was studied, and chronically, the effects of a 30% lactose diet fed from postnatal (PN) Day 15 onwards were evaluated. Intestinal lactase activity, as assessed both in vivo and in vitro, was compared between both test methods and diet groups (lactose vs. control). A 50%-75% decreased digestive capability towards lactose was observed from weaning into adulthood. Instillation of lactose in adult rats showed disproportionally low increases in plasma glucose levels and did not elicit an insulin response. However, gavages comprising maltodextrin gave rise to significant plasma glucose and insulin responses, indicative of a bias of the adult GI tract to digest glucose polymers. Despite the residual intestinal lactase activity shown, a 30% lactose diet was poorly digested by adult rats: the lactose diet rendered the animals less heavy and virtually devoid of body fat, whereas their cecum tripled in size, suggesting an increased bacterial fermentation. The observed acute and chronic effects of lactose exposure in adult rats cannot be explained by the residual intestinal lactase activity assessed.

  18. Variant Amino Acid Residues Alter the Enzyme Activity of Peanut Type 2 Diacylglycerol Acyltransferases

    Directory of Open Access Journals (Sweden)

    Ling Zheng

    2017-10-01

    Full Text Available Diacylglycerol acyltransferase (DGAT catalyzes the final step in triacylglycerol (TAG biosynthesis via the acyl-CoA-dependent acylation of diacylglycerol. This reaction is a major control point in the Kennedy pathway for biosynthesis of TAG, which is the most important form of stored metabolic energy in most oil-producing plants. In this study, Arachis hypogaea type 2 DGAT (AhDGAT2 genes were cloned from the peanut cultivar ‘Luhua 14.’ Sequence analysis of 11 different peanut cultivars revealed a gene family of 8 peanut DGAT2 genes (designated AhDGAT2a-h. Sequence alignments revealed 21 nucleotide differences between the eight ORFs, but only six differences result in changes to the predicted amino acid (AA sequences. A representative full-length cDNA clone (AhDGAT2a was characterized in detail. The biochemical effects of altering the AhDGAT2a sequence to include single variable AA residues were tested by mutagenesis and functional complementation assays in transgenic yeast systems. All six mutant variants retained enzyme activity and produced lipid droplets in vivo. The N6D and A26P mutants also displayed increased enzyme activity and/or total cellular fatty acid (FA content. N6D mutant mainly increased the content of palmitoleic acid, and A26P mutant mainly increased the content of palmitic acid. The A26P mutant grew well both in the presence of oleic and C18:2, but the other mutants grew better in the presence of C18:2. AhDGAT2 is expressed in all peanut organs analyzed, with high transcript levels in leaves and flowers. These levels are comparable to that found in immature seeds, where DGAT2 expression is most abundant in other plants. Over-expression of AhDGAT2a in tobacco substantially increased the FA content of transformed tobacco seeds. Expression of AhDGAT2a also altered transcription levels of endogenous tobacco lipid metabolic genes in transgenic tobacco, apparently creating a larger carbon ‘sink’ that supports increased FA

  19. Synthesis of high specific active tritiated Leu-enkephalin in the leucine residue

    Energy Technology Data Exchange (ETDEWEB)

    Baba, S.; Hasegawa, H.; Shinohara, Y. (Tokyo Coll. of Pharmacy (Japan))

    1989-12-01

    Leu-enkephalin labelled with tritium in the Leu residue has been prepared. Synthesis of the precursor peptide, (4,5-dehydroLeu{sup 5}-)Leu-enkephalin, was carried out by solid phase synthesis using Fmoc amino acid derivatives. The peptide was tritiated catalytically yielding {sup 3}H-Leu-enkephalin with a specific radioactivity of 4.39 TBq/mmol. The distribution of tritium label was investigated by reversed-phase high performance liquid chromatography with a synchronized accumulating radioisotope detector following acidic and enzymatic hydrolysis, which confirmed that the tritium label was entirely located at the Leu residue. (author).

  20. A comparative study on the modes of action of TAK-438, a novel potassium-competitive acid blocker, and lansoprazole in primary cultured rabbit gastric glands.

    Science.gov (United States)

    Matsukawa, Jun; Hori, Yasunobu; Nishida, Haruyuki; Kajino, Masahiro; Inatomi, Nobuhiro

    2011-05-01

    TAK-438 is a novel potassium-competitive acid blocker (P-CAB) type antisecretory agent that reversibly inhibits gastric H+, K+-ATPase. Previously, we showed that TAK-438 has superior efficacy compared to lansoprazole, a proton pump inhibitor, in the inhibition of acid secretion in vivo. In this study, we investigated the differences in the mode of actions of the two drugs using primary cultured rabbit gastric glands. TAK-438 and lansoprazole inhibited gastric acid formation in acutely isolated gastric glands (IC₅₀) values, 0.30 and 0.76 μM, respectively). In cultured gastric glands that were preincubated with TAK-438, the inhibitory effect on forskolin-stimulated acid formation was augmented over the incubation period, whereas the inhibitory effect of lansoprazole was not affected by time of incubation. Next, we evaluated the durations of the actions of TAK-438 and lansoprazole after gastric glands were incubated with either drug for 2h followed by washout. Even 8h after the drug washout, TAK-438 at higher concentrations inhibited acid formation, but the inhibitory effect of lansoprazole disappeared immediately after washout. Additionally, only a small amount of [¹⁴C] lansoprazole accumulated in resting glands, and this accumulation was enhanced by treatment with 1 μM of forskolin. In contrast, high levels of [¹⁴C] TAK-438 accumulated in both resting and forskolin-treated glands. Furthermore, a 2-h preincubation followed by washout demonstrated a slow clearance of [¹⁴C] TAK-438 from the glands. These findings suggest that TAK-438 exerts a longer and more potent antisecretory effect than lansoprazole as a result of its high accumulation and slow clearance from the gastric glands. Copyright © 2011 Elsevier Inc. All rights reserved.

  1. Substitution of Two Active-Site Residues Alters C9-Hydroxylation in a Class II Diterpene Synthase.

    Science.gov (United States)

    Mafu, Sibongile; Fischer, Emil; Addison, J Bennett; Riberio Barbosana, Isabel; Zerbe, Philipp

    2016-12-14

    Diterpenes form a vast and diverse class of natural products of both ecological and economic importance. Class II diterpene synthase (diTPS) enzymes control the committed biosynthetic reactions underlying diterpene chemical diversity. Homology modelling with site-directed mutagenesis identified two active-site residues in the horehound (Marrubium vulgare) class II diTPS peregrinol diphosphate synthase (MvCPS1); residue substitutions abolished the unique MvCPS1-catalysed water-capture reaction at C9 and redirected enzyme activity toward formation of an alternative product, halima-5(10),13-dienyl diphosphate. These findings contributed new insight into the steric interactions that govern diTPS-catalysed regiospecific oxygenation reactions and highlight the feasibility of diTPS engineering to provide a broader spectrum of bioactive diterpene natural products. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Determination of the crystal structure and active residues of FabV, the enoyl-ACP reductase from Xanthomonas oryzae.

    Directory of Open Access Journals (Sweden)

    He Li

    Full Text Available Enoyl-ACP reductase (ENR catalyses the last reduction reaction in the fatty acid elongation cycle in bacteria and is a good antimicrobial target candidate. FabV is the most recently discovered class of ENR, but we lack information about the atomic structure and the key residues involved in reductase activity except for the known conserved tyrosine and lysine residues in the Y-X(8-K active site motif.Here we report the crystal structure of FabV from Xanthomonas oryzae (xoFabV. The crystal structure of this enzyme has been solved to 1.6 Å resolution in space group P2(12(12(1. The model of xoFabV consists of one monomer in the asymmetric unit which is composed of 13 α-helices and 11 β-strands, representing a canonical Rossmann fold architecture. Structural comparison presents that the locations of the conserved tyrosine (Y236 and lysine (K245 residues in the Y-X(8-K active site motif of xoFabV and the Y-X(6-K motif of ecFabI are notably similar. However, the conformations of Y236 in xoFabV and Y156 in ecFabI are distinct. Structure-based site-directed mutagenesis and enzymatic activity assays reveal that in addition to the conserved Y236 and K245 in the Y-X(8-K motif, Y53, D111 and Y226 are key residues implicated in the reductase activity, and F113 and T276 are also important for enzyme function. Moreover, a proposed active lysine located immediately after the Y-X(8-K motif in FabV from Burkholderia mallei (bmFabV is altered to an inactive V246 in xoFabV.We determine the first crystal structure of the FabV enzyme and identify several residues important for its enzymatic activity. These findings lay a solid foundation for the development of specific antibacterial inhibitors of the pathogenic bacteria, such as Vibrio cholerae, Burkholderia species and Xanthomonas species.

  3. Physical activity and gastric residuals as biomarkers for region-specific NEC lesions in preterm neonates

    DEFF Research Database (Denmark)

    Cao, Muqing; Andersen, Anders Daniel; Li, Yanqi

    2016-01-01

    surveillance system and they were evaluated twice daily for clinical signs of apathy, discoloration, respiratory distress, abdominal distension and diarrhea. The volume of gastric residuals and the presence of macroscopic NEC-like lesions in the stomach, intestine and colon were recorded at euthanasia on day 5...

  4. Minimizing Actuator-Induced Residual Error in Active Space Telescope Primary Mirrors

    Science.gov (United States)

    2010-09-01

    modeling process using Matlab and MSC Nastran to sim- ulate actuator-induced residual error. . . . . . . . . . . . . . . . . . 47 3-3 Finite element mirror...automatically gener- ates the structural design of space telescope via Nastran , adds representative dynamic disturbances, simulates the application of...polynomials and Bessel functions. The authors employ a piezoelectrically- actuated membrane mirror model implemented using MSC Nastran to calculate the

  5. The conserved basic residues and the charged amino acid residues at the α-helix of the zinc finger motif regulate the nuclear transport activity of triple C2H2 zinc finger proteins

    Science.gov (United States)

    Lin, Chih-Ying

    2018-01-01

    Zinc finger (ZF) motifs on proteins are frequently recognized as a structure for DNA binding. Accumulated reports indicate that ZF motifs contain nuclear localization signal (NLS) to facilitate the transport of ZF proteins into nucleus. We investigated the critical factors that facilitate the nuclear transport of triple C2H2 ZF proteins. Three conserved basic residues (hot spots) were identified among the ZF sequences of triple C2H2 ZF proteins that reportedly have NLS function. Additional basic residues can be found on the α-helix of the ZFs. Using the ZF domain (ZFD) of Egr-1 as a template, various mutants were constructed and expressed in cells. The nuclear transport activity of various mutants was estimated by analyzing the proportion of protein localized in the nucleus. Mutation at any hot spot of the Egr-1 ZFs reduced the nuclear transport activity. Changes of the basic residues at the α-helical region of the second ZF (ZF2) of the Egr-1 ZFD abolished the NLS activity. However, this activity can be restored by substituting the acidic residues at the homologous positions of ZF1 or ZF3 with basic residues. The restored activity dropped again when the hot spots at ZF1 or the basic residues in the α-helix of ZF3 were mutated. The variations in nuclear transport activity are linked directly to the binding activity of the ZF proteins with importins. This study was extended to other triple C2H2 ZF proteins. SP1 and KLF families, similar to Egr-1, have charged amino acid residues at the second (α2) and the third (α3) positions of the α-helix. Replacing the amino acids at α2 and α3 with acidic residues reduced the NLS activity of the SP1 and KLF6 ZFD. The reduced activity can be restored by substituting the α3 with histidine at any SP1 and KLF6 ZFD. The results show again the interchangeable role of ZFs and charge residues in the α-helix in regulating the NLS activity of triple C2H2 ZF proteins. PMID:29381770

  6. The conserved basic residues and the charged amino acid residues at the α-helix of the zinc finger motif regulate the nuclear transport activity of triple C2H2 zinc finger proteins.

    Science.gov (United States)

    Lin, Chih-Ying; Lin, Lih-Yuan

    2018-01-01

    Zinc finger (ZF) motifs on proteins are frequently recognized as a structure for DNA binding. Accumulated reports indicate that ZF motifs contain nuclear localization signal (NLS) to facilitate the transport of ZF proteins into nucleus. We investigated the critical factors that facilitate the nuclear transport of triple C2H2 ZF proteins. Three conserved basic residues (hot spots) were identified among the ZF sequences of triple C2H2 ZF proteins that reportedly have NLS function. Additional basic residues can be found on the α-helix of the ZFs. Using the ZF domain (ZFD) of Egr-1 as a template, various mutants were constructed and expressed in cells. The nuclear transport activity of various mutants was estimated by analyzing the proportion of protein localized in the nucleus. Mutation at any hot spot of the Egr-1 ZFs reduced the nuclear transport activity. Changes of the basic residues at the α-helical region of the second ZF (ZF2) of the Egr-1 ZFD abolished the NLS activity. However, this activity can be restored by substituting the acidic residues at the homologous positions of ZF1 or ZF3 with basic residues. The restored activity dropped again when the hot spots at ZF1 or the basic residues in the α-helix of ZF3 were mutated. The variations in nuclear transport activity are linked directly to the binding activity of the ZF proteins with importins. This study was extended to other triple C2H2 ZF proteins. SP1 and KLF families, similar to Egr-1, have charged amino acid residues at the second (α2) and the third (α3) positions of the α-helix. Replacing the amino acids at α2 and α3 with acidic residues reduced the NLS activity of the SP1 and KLF6 ZFD. The reduced activity can be restored by substituting the α3 with histidine at any SP1 and KLF6 ZFD. The results show again the interchangeable role of ZFs and charge residues in the α-helix in regulating the NLS activity of triple C2H2 ZF proteins.

  7. Selective effects of charge on G protein activation by FSH-receptor residues 551-555 and 650-653.

    Science.gov (United States)

    Grasso, P; Deziel, M R; Reichert, L E

    1995-01-01

    Two cytosolic regions of the rat testicular FSH receptor (FSHR), residues 533-555 and 645-653, have been identified as G protein-coupling domains. We localized the activity in these domains to their C-terminal sequences, residues 551-555 (KIAKR, net charge +3) and 650-653 (RKSH, net charge +3), and examined the effects of charge on G protein activation by the C-terminal peptides, using synthetic analogs containing additions, through alanine (A) linkages, of arginine (R, +), histidine (H, +) or both. RA-KIAKR (net charge +4) mimicked the effect of FSHR-(551-555) on guanine nucleotide exchange in rat testis membranes, but reduced its ability to inhibit FSH-stimulated estradiol biosynthesis in cultured rat Sertoli cells. Further increasing net charge by the addition of H (HARA-KIAKR, net charge +5) increased guanosine 5'-triphosphate (GTP) binding, but eliminated FSHR-(551-555) effects on FSH-stimulated steroidogenesis. HA-RKSH (net charge +4) significantly inhibited guanine nucleotide exchange in rat testis membranes, but stimulated basal and potentiated FSH-induced estradiol biosynthesis in cultured rat Sertoli cells. Addition of two H residues (HAHA-RKSH, net charge +5) restored GTP binding and further potentiated basal and FSH-stimulated steroidogenesis. These results suggest that positive charges in G protein-coupling domains of the FSHR play a role in modulating G protein activation and postbinding effects of FSH, such as steroidogenesis.

  8. Tak v současných mediálních mluvených rozhovorech: ty zákony co se vztahujou teďkon na nás soukromníky tak to asi budu muset zavřít (1. část)

    Czech Academy of Sciences Publication Activity Database

    Jílková, Lucie

    -, č. 14 (2016), s. 12-33 ISSN 1804-137X R&D Projects: GA ČR GA15-01116S Institutional support: RVO:68378092 Keywords : word tak * DIALOG corpus * mass media dialogue * spoken language * meanings and functions of the Czech word tak Subject RIV: AI - Linguistics OBOR OECD: Linguistics

  9. Mutational analysis of amino acid residues involved in catalytic activity of a family 18 chitinase from tulip bulbs.

    Science.gov (United States)

    Suzukawa, Keisuke; Yamagami, Takeshi; Ohnuma, Takayuki; Hirakawa, Hideki; Kuhara, Satoru; Aso, Yoichi; Ishiguro, Masatsune

    2003-02-01

    We expressed chitinase-1 (TBC-1) from tulip bulbs (Tulipa bakeri) in E. coli cells and used site-directed mutagenesis to identify amino acid residues essential for catalytic activity. Mutations at Glu-125 and Trp-251 completely abolished enzyme activity, and activity decreased with mutations at Asp-123 and Trp-172 when glycolchitin was the substrate. Activity changed with the mutations of Trp-251 to one of several amino acids with side-chains of little hydrophobicity, suggesting that hydrophobic interaction of Trp-251 is important for the activity. Molecular dynamics (MD) simulation analysis with hevamine as the model compound showed that the distance between Asp-123 and Glu-125 was extended by mutation of Trp-251. Kinetic studies of Trp-251-mutated chitinases confirmed these various phenomena. The results suggested that Glu-125 and Trp-251 are essential for enzyme activity and that Trp-251 had a direct role in ligand binding.

  10. Estimates of biomass in logging residue and standing residual inventory following tree-harvest activity on timberland acres in the southern region

    Science.gov (United States)

    Roger C. Conner; Tony G. Johnson

    2011-01-01

    This report provides estimates of biomass (green tons) in logging residue and standing residual inventory on timberland acres with evidence of tree cutting. Biomass as defined by Forest Inventory and Analysis is the aboveground dry weight of wood in the bole and limbs of live trees ≥ 1-inch diameter at breast height (d.b.h.), and excludes tree foliage, seedlings, and...

  11. Modulating the pH-activity profile of cellulase A from Cellulomonas fimi by replacement of surface residues.

    Science.gov (United States)

    Cockburn, Darrell W; Clarke, Anthony J

    2011-05-01

    One industrial process for the production of cellulosic ethanol and or value-added products involves exposing the cellulose content of plant materials by steam explosion in the presence of strong acid, followed by its neutralization and subsequent digestion with a cocktail of cellulolytic enzymes. These enzymes typically have activity optima at slightly acidic or neutral pH and so generating enzymes that are more active and tolerant in more acidic conditions would help to reduce associated costs. Here, we describe the engineering of cellulase A from Cellulomonas fimi as a model to replace residues that were identified as potentially influencing the pH-activity profile of the enzyme based on sequence alignments and analysis of the known three-dimensional structures of other CAZy family 6 glycoside hydrolases with the aim to lower its pH optimum. Twelve specific residues and a sequence of eight were identified and a total of 30 mutant enzymes were generated. In addition to being replaced with natural amino acids, some of the identified residues were substituted with cysteine and subsequently oxidized to cysteinesulfinate. Of the four single amino acid replacements that produced enhancements of activity at acidic pH, three involved the removal of charged groups from the surface of the enzyme. The generation of double mutations provided mixed results but the combination of Glu407 → Ala and Tyr321 → Phe replacements had an additive effect on the enhancement, reaching a total activity that was 162% of the wild-type level. This study thus illustrated the utility of altering the surface charge properties of the family 6 glycoside hydrolases to enhance activity at low pH and thereby an avenue for further protein engineering.

  12. Role of cysteine residues in the structure, stability, and alkane producing activity of cyanobacterial aldehyde deformylating oxygenase.

    Directory of Open Access Journals (Sweden)

    Yuuki Hayashi

    Full Text Available Aldehyde deformylating oxygenase (AD is a key enzyme for alkane biosynthesis in cyanobacteria, and it can be used as a catalyst for alkane production in vitro and in vivo. However, three free Cys residues in AD may impair its catalytic activity by undesired disulfide bond formation and oxidation. To develop Cys-deficient mutants of AD, we examined the roles of the Cys residues in the structure, stability, and alkane producing activity of AD from Nostoc punctiforme PCC 73102 by systematic Cys-to-Ala/Ser mutagenesis. The C71A/S mutations reduced the hydrocarbon producing activity of AD and facilitated the formation of a dimer, indicating that the conserved Cys71, which is located in close proximity to the substrate-binding site, plays crucial roles in maintaining the activity, structure, and stability of AD. On the other hand, mutations at Cys107 and Cys117 did not affect the hydrocarbon producing activity of AD. Therefore, we propose that the C107A/C117A double mutant is preferable to wild type AD for alkane production and that the double mutant may be used as a pseudo-wild type protein for further improvement of the alkane producing activity of AD.

  13. The roles of selected arginine and lysine residues of TAFI (Pro-CPU) in its activation to TAFIa by the thrombin-thrombomodulin complex.

    Science.gov (United States)

    Wu, Chengliang; Kim, Paul Y; Manuel, Reg; Seto, Marian; Whitlow, Marc; Nagashima, Mariko; Morser, John; Gils, Ann; Declerck, Paul; Nesheim, Michael E

    2009-03-13

    Thrombomodulin (TM) increases the catalytic efficiency of thrombin (IIa)-mediated activation of thrombin-activable fibrinolysis inhibitor (TAFI) 1250-fold. Negatively charged residues of the C-loop of TM-EGF-like domain 3 are required for TAFI activation. Molecular models suggested several positively charged residues of TAFI with which the C-loop residues could interact. Seven TAFI mutants were constructed to determine if these residues are required for efficient TAFI activation. TAFI wild-type or mutants were activated in the presence or absence of TM and the kinetic parameters of TAFI activation were determined. When the three consecutive lysine residues in the activation peptide of TAFI were substituted with alanine (K42/43/44A), the catalytic efficiencies for TAFI activation with TM decreased 8-fold. When other positively charged surface residues of TAFI (Lys-133, Lys-211, Lys-212, Arg-220, Lys-240, or Arg-275) were mutated to alanine, the catalytic efficiencies for TAFI activation with TM decreased by 1.7-2.7-fold. All decreases were highly statistically significant. In the absence of TM, catalytic efficiencies ranged from 2.8-fold lower to 1.24-fold higher than wild-type. None of these, except the 2.8-fold lower value, was statistically significant. The average half-life of the TAFIa mutants was 8.1+/-0.6 min, and that of wild type was 8.4+/-0.3 min at 37 degrees C. Our data show that these residues are important in the activation of TAFI by IIa, especially in the presence of TM. Whether the mutated residues promote a TAFI-TM or TAFI-IIa interaction remains to be determined. In addition, these residues do not influence spontaneous inactivation of TAFIa.

  14. Aromatic residue position on the nonpolar face of class a amphipathic helical peptides determines biological activity.

    Science.gov (United States)

    Datta, Geeta; Epand, Raquel F; Epand, Richard M; Chaddha, Manjula; Kirksey, Matthew A; Garber, David W; Lund-Katz, Sissel; Phillips, Michael C; Hama, Susan; Navab, Mohamad; Fogelman, Alan M; Palgunachari, Mayakonda N; Segrest, Jere P; Anantharamaiah, G M

    2004-06-18

    The apolipoprotein A-I mimetic peptide 4F (Ac-DWFKAFYDKVAEKFKEAF-NH(2)), with four Phe residues on the nonpolar face of the amphipathic alpha-helix, is strongly anti-inflammatory, whereas two 3F analogs (3F(3) and 3F(14)) are not. To understand how changes in helix nonpolar face structure affect function, two additional 3F analogs, Ac-DKLKAFYDKVFEWAKEAF-NH(2) (3F-1) and Ac-DKWKAVYDKFAEAFKEFL-NH(2) (3F-2), were designed using the same amino acid composition as 3F(3) and 3F(14). The aromatic residues in 3F-1 and 3F-2 are near the polar-nonpolar interface and at the center of the nonpolar face of the helix, respectively. Like 4F, but in contrast to 3F(3) and 3F(14), these peptides effectively inhibited lytic peptide-induced hemolysis, oxidized phospholipid-induced monocyte chemotaxis, and scavenged lipid hydroperoxides from low density lipoprotein. High pressure liquid chromatography retention times and monolayer exclusion pressures indicated that there is no direct correlation of peptide function with lipid affinity. Fluorescence studies suggested that, although the peptides bind phospholipids similarly, the Trp residue in 4F, 3F-1, and 3F-2 is less motionally restricted than in 3F(3) and 3F(14). Based on these results and molecular modeling studies, we propose that the arrangement of aromatic residues in class A amphipathic helical molecules regulates entry of reactive oxygen species into peptide-phospholipid complexes, thereby reducing the extent of monocyte chemotaxis, an important step in atherosclerosis.

  15. The effects of the surface-exposed residues on the binding and hydrolytic activities of Vibrio carchariae chitinase A

    Directory of Open Access Journals (Sweden)

    Suginta Wipa

    2008-01-01

    Full Text Available Abstract Background Vibrio carchariae chitinase A (EC3.2.1.14 is a family-18 glycosyl hydrolase and comprises three distinct structural domains: i the amino terminal chitin binding domain (ChBD; ii the (α/β8 TIM barrel catalytic domain (CatD; and iii the α + β insertion domain. The predicted tertiary structure of V. carchariae chitinase A has located the residues Ser33 & Trp70 at the end of ChBD and Trp231 & Tyr245 at the exterior of the catalytic cleft. These residues are surface-exposed and presumably play an important role in chitin hydrolysis. Results Point mutations of the target residues of V. carchariae chitinase A were generated by site-directed mutagenesis. With respect to their binding activity towards crystalline α-chitin and colloidal chitin, chitin binding assays demonstrated a considerable decrease for mutants W70A and Y245W, and a notable increase for S33W and W231A. When the specific hydrolyzing activity was determined, mutant W231A displayed reduced hydrolytic activity, whilst Y245W showed enhanced activity. This suggested that an alteration in the hydrolytic activity was not correlated with a change in the ability of the enzyme to bind to chitin polymer. A mutation of Trp70 to Ala caused the most severe loss in both the binding and hydrolytic activities, which suggested that it is essential for crystalline chitin binding and hydrolysis. Mutations varied neither the specific hydrolyzing activity against pNP-[GlcNAc]2, nor the catalytic efficiency against chitohexaose, implying that the mutated residues are not important in oligosaccharide hydrolysis. Conclusion Our data provide direct evidence that the binding as well as hydrolytic activities of V. carchariae chitinase A to insoluble chitin are greatly influenced by Trp70 and less influenced by Ser33. Though Trp231 and Tyr245 are involved in chitin hydrolysis, they do not play a major role in the binding process of crystalline chitin and the guidance of the chitin chain

  16. An atypical residue in the pore of Varroa destructor GABA-activated RDL receptors affects picrotoxin block and thymol modulation.

    Science.gov (United States)

    Price, Kerry L; Lummis, Sarah C R

    2014-12-01

    GABA-activated RDL receptors are the insect equivalent of mammalian GABAA receptors, and play a vital role in neurotransmission and insecticide action. Here we clone the pore lining M2 region of the Varroa mite RDL receptor and show that it has 4 atypical residues when compared to M2 regions of most other insects, including bees, which are the major host of Varroa mites. We create mutant Drosophila RDL receptors containing these substitutions and characterise their effects on function. Using two electrode voltage clamp electrophysiology we show that one substitution (T6'M) ablates picrotoxin inhibition and increases the potency of GABA. This mutation also alters the effect of thymol, which enhances both insect and mammalian GABA responses, and is widely used as a miticide. Thymol decreases the GABA EC50 of WT receptors, enhancing responses, but in T6'M-containing receptors it is inhibitory. The other 3 atypical residues have no major effects on either the GABA EC50, the picrotoxin potency or the effect of thymol. In conclusion we show that the RDL 6' residue is important for channel block, activation and modulation, and understanding its function also has the potential to prove useful in the design of Varroa-specific insecticidal agents. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  17. Contact and leaf residue activity of insecticides against the sweet Corn pest Euxesta stigmatias (Diptera: Otitidae).

    Science.gov (United States)

    Nuessly, Gregg S; Hentz, Matthew G

    2004-04-01

    Damage by Euxesta stigmatias Loew larvae to sweet corn renders the ears unmarketable. This report evaluates the efficacy of insecticides labeled for armyworm control in Florida sweet corn against E. stigmatias adults. Tests were performed in controlled settings by using direct contact and dried plant residues of esfenvalerate, cyfluthrin, lambda-cyhalothrin, chlorpyrifos, methyl parathion, methomyl, and thiodicarb. Direct application of all insecticides except thiodicarb quickly killed or caused incapacitating sublethal affects (uncoordinated movement, uncontrolled twitching, and hyperextension of mouthparts and ovipositor) to > 75% of the flies. Low rates (0.56 kg [AI]/ha) of chlorpyrifos and methyl parathion provided the most efficient control, killing 100 and 93%, respectively, within 2 h of direct contact. Low rates of pyrethroids induced low mortality but high sublethal effects that together immobilized nearly 100% of adults within 1 h of exposure. Mortality reached 95% within 2 h of direct contact in flies treated with high rates of pyrethroids. Methomyl killed as many as 94%, but required 24 h to reach this level after direct treatment. Residues on dipped leaves and field-treated plants of all tested insecticides except methyl parathion were less effective at killing adults compared with direct contact tests. Pyrethroid residues (particularly cyfluthrin) on field planted sweet corn induced significantly higher levels of sublethal effects (57-70%), and for a longer period of time, compared with materials in the other classes of chemistry.

  18. Reconstructing late Neolithic plant economies at the Eastern Adriatic site of Velištak (5th millennium cal BC

    Directory of Open Access Journals (Sweden)

    Kelly Reed

    2016-12-01

    Full Text Available The archaeobotanical remains from Velištak are the first evidence of plant economies from an open-air settlement dating to the late Neolithic Hvar culture in Croatia (c. 4900–4000 cal BC. The results presented here are from the 2007–2013 field seasons. Based on an examination of carbonised macro-remains, it is suggested that emmer, einkorn, and barley were the main crops at Velištak, along with lentils, bitter vetch, and possibly peas and flax. Wild plants were also exploited, with evidence of wild fruits, such as cornelian cherry. Similarities with archaeobotanical finds from the early/middle Neolithic (c. 6000–4900 cal BC also suggest that plant economies remained relatively unchanged during the Neolithic.

  19. Contribution of factor VIII light-chain residues 2007-2016 to an activated protein C-interactive site.

    Science.gov (United States)

    Takeyama, Masahiro; Wakabayashi, Hironao; Fay, Philip J

    2013-02-01

    Although factor (F) VIIIa is inactivated by activated protein C (APC) through cleavages in the FVIII heavy chain-derived A1 (Arg(336)) and A2 subunits (Arg(562), the FVIII light chain (LC) contributes to catalysis by binding the enzyme. ELISA-based binding assays showed that FVIII and FVIII LC bound to immobilised active site-modified APC (DEGR-APC) (apparent K(d) ~270 nM and 1.0 μM, respectively). Fluid-phase binding studies using fluorescence indicated an estimated K(d) of ~590 nM for acrylodan-labelled LC binding to DEGR-APC. Furthermore, FVIII LC effectively competed with FVIIIa in blocking APC-catalysed cleavage at Arg(336) (K(i) = 709 nM). A binding site previously identified near the C-terminal end of the A3 domain (residues 2007-2016) of FVIII LC was subjected to Ala-scanning mutagenesis. FXa generation assays and western and dot blotting were employed to assess the contribution of these residues to FVIIIa interactions with APC. Virtually all variants tested showed reductions in the rates of APC-catalysed inactivation of the cofactor and cleavage at the primary inactivation site (Arg(336)), with maximal reductions in inactivation rates (~3-fold relative to WT) and cleavage rates (~3 to ~9-fold relative to WT) observed for the Met2010Ala, Ser2011Ala, and Leu2013Ala variants. Titration of FVIIIa substrate concentration monitoring cleavage by a dot blot assay indicated that these variants also showed ~3-fold increases relative to WT while a double mutant (Met2010Ala/Ser2011Ala) showed a >4-fold increase in K(m). These results show a contribution of a number of residues within the 2007-2016 sequence, and in particular residues Met2010, Ser2011, and Leu2013 to an APC-interactive site.

  20. The Arabidopsis KS-type dehydrin recovers lactate dehydrogenase activity inhibited by copper with the contribution of His residues.

    Science.gov (United States)

    Hara, Masakazu; Monna, Shuhei; Murata, Takae; Nakano, Taiyo; Amano, Shono; Nachbar, Markus; Wätzig, Hermann

    2016-04-01

    Dehydrin, which is one of the late embryogenesis abundant (LEA) proteins, is involved in the ability of plants to tolerate the lack of water. Although many reports have indicated that dehydrins bind heavy metals, the physiological role of this metal binding has not been well understood. Here, we report that the Arabidopsis KS-type dehydrin (AtHIRD11) recovered the lactate dehydrogenase (LDH) activity denatured by Cu(2+). The LDH activity was partially inhibited by 0.93 μM Cu(2+) but totally inactivated by 9.3 μM Cu(2+). AtHIRD11 recovered the activity of LDH treated with 9.3 μM Cu(2+) in a dose-dependent manner. The recovery activity of AtHIRD11 was significantly higher than those of serum albumin and lysozyme. The conversion of His residues to Ala in AtHIRD11 resulted in the loss of the Cu(2+) binding of the protein as well as the disappearance of the conformational change induced by Cu(2+) that is observed by circular dichroism spectroscopy. The mutant protein showed lower recovery activity than the original AtHIRD11. These results indicate that AtHIRD11 can reactivate LDH inhibited by Cu(2+) via the His residues. This function may prevent physiological damage to plants due to heavy-metal stress. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  1. Türk ve Yabancı Futbol Takımlarının Borsa Performansları Üzerine Bir Uygulama

    Directory of Open Access Journals (Sweden)

    Feyyaz ZEREN

    2013-09-01

    Full Text Available Bu çalışmanın amacı hisse senetleri borsada işlem gören yerli ve yabancı toplam 8 futbol takımının kendi liglerinde aldıkları maç sonuçları ile borsa performansları arasında bir ilişki olup olmadığını araştırmaktadır. Türk futbol takımlarını Beşiktaş, Galatasaray, Fenerbahçe ve Trabzonspor; yabancı takımları ise Roma, Borussia Dortmund, Olympique Lyon ve Juventus oluşturmaktadır. Çalışmada bu takımların 2007-2011 sezonlarında oynadıkları maçların sonuçları temel alınmıştır. Bu amaca ulaşmak üzere SPSS 15 programında T Testi uygulaması yapılmıştır. Yapılan analiz sonucunda 8 takım içerisinden aldığı galibiyetler ile borsa performansı arasında anlamlı ilişki tespit edilen tek takım Galatasaray olarak çıkmıştır. Mağlubiyetler ile borsa performansı arasında anlamlı ilişki tespit edilen takımlar ise Fenerbahçe, Juventus, Roma ve Borussia Dortmund takımlarıdır.

  2. A remote palm domain residue of RB69 DNA polymerase is critical for enzyme activity and influences the conformation of the active site.

    Directory of Open Access Journals (Sweden)

    Agata Jacewicz

    Full Text Available Non-conserved amino acids that are far removed from the active site can sometimes have an unexpected effect on enzyme catalysis. We have investigated the effects of alanine replacement of residues distant from the active site of the replicative RB69 DNA polymerase, and identified a substitution in a weakly conserved palm residue (D714A, that renders the enzyme incapable of sustaining phage replication in vivo. D714, located several angstroms away from the active site, does not contact the DNA or the incoming dNTP, and our apoenzyme and ternary crystal structures of the Pol(D714A mutant demonstrate that D714A does not affect the overall structure of the protein. The structures reveal a conformational change of several amino acid side chains, which cascade out from the site of the substitution towards the catalytic center, substantially perturbing the geometry of the active site. Consistent with these structural observations, the mutant has a significantly reduced k pol for correct incorporation. We propose that the observed structural changes underlie the severe polymerization defect and thus D714 is a remote, non-catalytic residue that is nevertheless critical for maintaining an optimal active site conformation. This represents a striking example of an action-at-a-distance interaction.

  3. A survey of ectoparasitic arthropods on domestic animals in Tak Province, Thailand.

    Science.gov (United States)

    Changbunjong, Tanasak; Buddhirongawatr, Ruangrat; Suwanpakdee, Sarin; Siengsanan, Jarunee; Yongyuttawichai, Plern; Cheewajorn, Kecha; Jangjaras, Juthathip; Sangloung, Charoonluk; Ratanakorn, Parntep

    2009-05-01

    In July 2008 a survey of ectoparasites on domestic animals was conducted in the Royal Thai Army areas of operation along the Thai-Myanmar Border, Tak Province, Thailand. Eleven different ectoparasites were collected: two species of hard ticks (Ixodidae), three species of fleas (Siphonaptera) and 6 species of sucking or chewing lice (2 species each in the suborders Anoplura, Ischnocera and Amblycera) were collected. Domestic dogs (Canis lupusfamiliaris) (n = 94) were found infested with 2 species of flea Ctenocephalides felis orientis (86.2%) and Echidnophaga gallinacea (1.1%), one species of tick, Rhipicephalus sanguineus (21.3%), and one louse species, Heterodoxus spiniger (7.4%). Domestic cats (Felis catus) (n = 6) were found infested with only flea species, Ctenocephalides felis felis (100%) and E. gallinacea (33.3%). Cattle (Bos indicus) (n = 11) had Rhipicephalus (Boophilus) microplus (72.7%), Linognathus vituli (27.3%), and Solenopotes capillatus (9.1%) present, while chickens (Gallus domesticus) (n = 10) had infestations with E. gallinacea (20%), and 3 lice species, Lipeurus caponis (10%), Goniodes dissimilis (10%) and Menopon gallinae (60%). This is believed to be the first report of S. capillatus collected in Thailand.

  4. The Study of Critical Thinking skills of Seventh Grade Hmong Hilltribe Students in Tak Province

    Directory of Open Access Journals (Sweden)

    Kritsana Lohakarok

    2017-06-01

    Full Text Available The purpose of this study was to study critical thinking skills of Hmong hilltribe students who were studying in seventh grade, in Tak province of Thailand. The research instrument used in this study was the critical thinking skills test developed by researcher. The test was a multiple-choice test which composed of 30 items with 30 total scores including three compositions of critical thinking skills ; 1 problem defining 2 decision making and data processing and 3 making conclusion. The participant in this study were 30 Hmong hilltribe students using purposive sampling who were studying in seventh grade in academic year 2015. The findings of the study revealed that the mean score of critical thinking skills of Hmong hilltribe students equaled 13.07. The maximum score was 20. The minimum score was 7 with the standard deviation 3.59. Moreover, the levels of critical thinking skills presented that 13.33% (n=4 of these students had critical thinking skills in a fair level, 23.33% (n=7 in a poor level and 63.33% (n=19 in a very poor level. In addition, considering separately in each composition of critical thinking skills, the results found that 60.24% of students got the right answers in decision making and data processing, 34.44% in problem defining and 25.67% in making conclusion, respectively.

  5. Salvador Dali’nin Takı Çalışmaları / Jewelry Works of Salvador Dali

    OpenAIRE

    Yağmur, Önder; Bulat, Serap

    2014-01-01

    Öz: Modern sanatın büyük isimlerinden olan Salvador Dali’nin resim sanatındaki dehasınınyanında takı sanatında da nasıl bir dahi olduğunun anlatılması gerekir. Sinemadantakıya kadar birçok sanatsal alanda çalışmalar ortaya koymuş olan Dali, kendisineözgü sanatsal üslubunu çalışmış olduğu her sanat dalında olduğu gibi takı eserlerine deyansıtmıştır. Salvador Dali’nin resimlerinde görülen detaycı ve zihni zorlayıcı özelliği,Dali’nin takı alanında ortaya koymuş olduğu eserlerinde de görülmektedi...

  6. Activation of the Torpedo nicotinic acetylcholine receptor. The contribution of residues alphaArg55 and gammaGlu93.

    Science.gov (United States)

    Kapur, Ankur; Davies, Martin; Dryden, William F; Dunn, Susan M J

    2006-03-01

    The Torpedo nicotinic acetylcholine receptor is a heteropentamer (alpha2betagammadelta) in which structurally homologous subunits assemble to form a central ion pore. Viewed from the synaptic cleft, the likely arrangement of these subunits is alpha-gamma-alpha-delta-beta lying in an anticlockwise orientation. High affinity binding sites for agonists and competitive antagonists have been localized to the alpha-gamma and alpha-delta subunit interfaces. We investigated the involvement of amino acids lying at an adjacent interface (gamma-alpha) in receptor properties. Recombinant Torpedo receptors, expressed in Xenopus oocytes, were used to investigate the consequences of mutating alphaArg55 and gammaGlu93, residues that are conserved in most species of the peripheral nicotinic receptors. Based on homology modeling, these residues are predicted to lie in close proximity to one another and it has been suggested that they may form a salt bridge in the receptor's three-dimensional structure (Sine et al. 2002 J Biol Chem277, 29 210-29 223). Although substitution of alphaR55 by phenylalanine or tryptophan resulted in approximately a six-fold increase in the EC50 value for acetylcholine activation, the charge reversal mutation (alphaR55E) had no significant effect. In contrast, the replacement of gammaE93 by an arginine conferred an eight-fold increase in the potency for acetylcholine-induced receptor activation. In the receptor carrying the double mutations, alphaR55E-gammaE93R or alphaR55F-gammaE93R, the potency for acetylcholine activation was partially restored to that of the wild-type. The results suggest that, although individually these residues influence receptor activation, direct interactions between them are unlikely to play a major role in the stabilization of different conformational states of the receptor.

  7. Radiation dosimetry by neutron or X ray fluorescence activation of residual silver in ionographic emulsions

    International Nuclear Information System (INIS)

    Heilmann, C.

    1987-01-01

    A global measuring technique which is sensitive enough to detect small silver contents in films for dosimetry applications is presented. The applications studied are neutron dosimetry by measuring residual silver due to recoil protons in developed emulsions and high dose dosimetry by the detection of photolytic silver in fixed emulsions. An individual fast neutron dosimeter which can be used in radiation protection was developed, along with an automatic data analysis and readout system. Application of this technique to the measurement of high radiation doses (100 to 1 million Gy) via the measurement of photolytic silver in fixed, but undeveloped, emulsions confirms the usefulness of the method [fr

  8. Preparation of Ecofriendly Formulations Containing Biologically Active Monoterpenes with Their Fumigant and Residual Toxicities against Adults of Culex pipiens

    Directory of Open Access Journals (Sweden)

    Mohamed E. I. Badawy

    2016-01-01

    Full Text Available Different mixtures of monoterpenes (ketone, alcohol, and alkene were loaded on paper discs and wax and their knockdown activities were evaluated against Culex pipiens adults. Some individual monoterpenes were also evaluated by residual toxicity technique. Citronella oil as a reference was also loaded separately or in combination with monoterpenes on paper discs and wax. The ketone monoterpenes mixture (camphor, menthone, carvone, and fenchone on paper discs was the most active (KT50 = 17.20 min followed by ketone monoterpenes with citronella oil (KT50 = 20.79 min and citronella oil alone (KT50 = 28.72 min. Wax formulations proved that the ketone and alcohol (geraniol, thymol, and menthol monoterpenes gave the most activity as knockdown (KT50 = 31.79 and 43.39 min, resp.. Alcohol monoterpenes formulation recorded KT50 = 43.39 min. Residual activity of tested individual monoterpenes reported that the menthol was more toxic than camphor and camphene. Generally, this study suggests that the monoterpenes have the properties, which make them used as eco-friendly compounds in the control programs of Cx. pipiens adult. The use of paper discs is more applicable than wax in the adulticidal formulations.

  9. Preparation of Ecofriendly Formulations Containing Biologically Active Monoterpenes with Their Fumigant and Residual Toxicities against Adults ofCulex pipiens.

    Science.gov (United States)

    Badawy, Mohamed E I; Taktak, Nehad E M; Awad, Osama M; Elfiki, Souraya A; Abou El-Ela, Nadia E

    2016-01-01

    Different mixtures of monoterpenes (ketone, alcohol, and alkene) were loaded on paper discs and wax and their knockdown activities were evaluated against Culex pipiens adults. Some individual monoterpenes were also evaluated by residual toxicity technique. Citronella oil as a reference was also loaded separately or in combination with monoterpenes on paper discs and wax. The ketone monoterpenes mixture (camphor, menthone, carvone, and fenchone) on paper discs was the most active (KT 50 = 17.20 min) followed by ketone monoterpenes with citronella oil (KT 50 = 20.79 min) and citronella oil alone (KT 50 = 28.72 min). Wax formulations proved that the ketone and alcohol (geraniol, thymol, and menthol) monoterpenes gave the most activity as knockdown (KT 50 = 31.79 and 43.39 min, resp.). Alcohol monoterpenes formulation recorded KT 50 = 43.39 min. Residual activity of tested individual monoterpenes reported that the menthol was more toxic than camphor and camphene. Generally, this study suggests that the monoterpenes have the properties, which make them used as eco-friendly compounds in the control programs of Cx. pipiens adult. The use of paper discs is more applicable than wax in the adulticidal formulations.

  10. Insights into the roles of non-catalytic residues in the active site of a GH10 xylanase with activity on cellulose.

    Science.gov (United States)

    Chu, Yindi; Tu, Tao; Penttinen, Leena; Xue, Xianli; Wang, Xiaoyu; Yi, Zhuolin; Gong, Li; Rouvinen, Juha; Luo, Huiying; Hakulinen, Nina; Yao, Bin; Su, Xiaoyun

    2017-11-24

    Bifunctional glycoside hydrolases have potential for cost-savings in enzymatic decomposition of plant cell wall polysaccharides for biofuels and bio-based chemicals. The N-terminal GH10 domain of a bifunctional multimodular enzyme Cb Xyn10C/Cel48B from Caldicellulosiruptor bescii is an enzyme able to degrade xylan and cellulose simultaneously. However, the molecular mechanism underlying its substrate promiscuity has not been elucidated. Herein, we discovered that the binding cleft of Cb Xyn10C would have at least six sugar-binding subsites by using isothermal titration calorimetry analysis of the inactive E140Q/E248Q mutant with xylo- and cello-oligosaccharides. This was confirmed by determining the catalytic efficiency of the wild-type enzyme on these oligosaccharides. The free form and complex structures of Cb Xyn10C with xylose- or glucose-configured oligosaccharide ligands were further obtained by crystallographic analysis and molecular modeling and docking. Cb Xyn10C was found to have a typical (β/α) 8 -TIM barrel fold and "salad-bowl" shape of GH10 enzymes. In complex structures with xylo-oligosaccharides, seven sugar-binding subsites were found, and many residues responsible for substrate interactions were identified. Site-directed mutagenesis indicated that 6 and 10 amino acid residues were key residues for xylan and cellulose hydrolysis, respectively. The most important residues are centered on subsites -2 and -1 near the cleavage site, whereas residues playing moderate roles could be located at more distal regions of the binding cleft. Manipulating the residues interacting with substrates in the distal regions directly or indirectly improved the activity of Cb Xyn10C on xylan and cellulose. Most of the key residues for cellulase activity are conserved across GH10 xylanases. Revisiting randomly selected GH10 enzymes revealed unreported cellulase activity, indicating that the dual function may be a more common phenomenon than has been expected. © 2017 by

  11. Bacillus subtilis with endocellulase and exocellulase activities isolated in the thermophilic phase from composting with coffee residues.

    Science.gov (United States)

    Siu-Rodas, Yadira; Calixto-Romo, María de Los Angeles; Guillén-Navarro, Karina; Sánchez, José E; Zamora-Briseño, Jesús Alejandro; Amaya-Delgado, Lorena

    2017-12-27

    The goal of this study was to isolate, select and characterize bacteria with cellulolytic activity from two different coffee residue composting piles, one of which had an internal temperature of 57°C and pH 5.5 and the other, a temperature of 61°C, and pH 9.3. Culture media were manipulated with carboxymethylcellulose and crystalline cellulose as sole carbon sources. The enzyme activity was assessed by hydrolysis halo formation, reducing sugar production and zymograms. Three out of twenty isolated strains showed higher enzymatic activity and were identified as Bacillus subtilis according to their morphological, physiological, biochemical characteristics and based on the sequence analysis of 16S rDNA regions. The enzymatic extracts of the three selected strains showed exocellulase and endocellulase maximum activity of 0.254 and 0.519 U/ml, respectively; the activity of these enzymes was maintained even in acid pH (4.8) and basic (9.3) and at temperatures of up to 60°C. The enzymatic activities observed in this study are within the highest reported for cellulose produced by bacteria of the genus Bacillus. Endocellulase activity was shown in the zymograms from 24h until 144h of incubation. Furthermore, the pH effect on the endocellulase activity is reported for the first time by zymograms. The findings in this study entail the possibility to use these enzymes in the procurement of fermentable substrates for the production of energy from the large amount of residues generated by the coffee agroindustry. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  12. Atividade residual de diuron, oxyfluorfen e prometryne no controle de Euphorbia heterophylla Residual activity of diuron, oxyfluorfen, and prometryne for Euphorbia heterophylla control

    OpenAIRE

    R.S. Oliveira Jr.; J.C. Carneiro; J. Constantin; G. Santos; A.C. Francischini; A.M. Oliveira Neto

    2012-01-01

    As aplicações de herbicidas em pré-emergência têm por finalidade a obtenção da atividade residual no início do ciclo das culturas. Assim, o objetivo deste trabalho foi avaliar a atividade residual dos herbicidas diuron, oxyfluorfen e prometryne, aplicados isoladamente ou em misturas, no controle de Euphorbia heterophylla. Oito experimentos foram conduzidos em casa de vegetação, aplicando-se doses dos herbicidas ou das misturas aos 30, 20, 10 e 0 dias antes da semeadura da planta daninha (DAS)...

  13. Roles of the C-terminal tyrosine residues of LAT in GPVI-induced platelet activation: insights into the mechanism of PLC gamma 2 activation.

    Science.gov (United States)

    Ragab, Ashraf; Séverin, Sonia; Gratacap, Marie-Pierre; Aguado, Enrique; Malissen, Marie; Jandrot-Perrus, Martine; Malissen, Bernard; Ragab-Thomas, Jeannie; Payrastre, Bernard

    2007-10-01

    Linker for activation of T cells (LAT) is an adaptor protein required for organization of the signaling machinery downstream of the platelet collagen receptor, the glycoprotein VI (GPVI). Here, we investigated the effect of LAT mutations on specific signaling pathways and on platelet functions in response to GPVI triggering by convulxin (Cvx). Using mice containing tyrosine to phenylalanine mutations of the adaptor, we show the crucial role played by the tyrosine residues at positions 175, 195, and 235 in the phosphorylation of LAT and in the whole pattern of protein tyrosine phosphorylation in response to Cvx. These 3 C-terminal tyrosine residues are important to recruit the tyrosine kinase Fyn, which may be involved in LAT phosphorylation. Efficient phosphoinositide 3-kinase (PI3K) activation requires the 3 C-terminal tyrosine residues of LAT but not its tyrosine 136. Interestingly, single mutation of the tyrosine 136 results in the loss of phospholipase C gamma2 (PLCgamma2) activation without affecting its PI3K-dependent membrane association, and is sufficient to impair platelet responses to Cvx. Thus, activation of PLCgamma2 via GPVI is dependent on 2 complementary events: its interaction with the tyrosine 136 of LAT and its membrane location, which itself requires events mediated by the 3 C-terminal tyrosines of LAT.

  14. Control of the activity of Rubisco from Chlamydomonas reinhardtii through the redox state of its cysteine residues

    OpenAIRE

    Sudhani, Hemanth P. K.

    2012-01-01

    RESUMEN de la MEMORIA de TESIS DOCTORAL titulada: CONTROL OF THE ACTIVITY OF RUBISCO FROM CHLAMYDOMONAS REINHARDTII THROUGH THE REDOX STATE OF ITS CYSTEINE RESIDUES. presentada por HEMANTH PHANI KUMAR SUDHANI para optar al título de DOCTOR dentro del Programa Oficial de Posgrado en BIOTECNOLOGÍA (D030-01) de la Universitat de València. Tema: La actividad del enzima fijador de carbono en eucariotas fotosintéticos, la ribulosa 1,5-bisfosfato carboxilas/oxigenasa (Rubisco), puede re...

  15. High Residual Activity of PMM2 in Patients’ Fibroblasts: Possible Pitfall in the Diagnosis of CDG-Ia (Phosphomannomutase Deficiency)

    OpenAIRE

    Grünewald, Stephanie; Schollen, Els; Van Schaftingen, Emile; Jaeken, Jaak; Matthijs, Gert

    2001-01-01

    Congenital disorders of glycosylation (CDGs) are a rapidly enlarging group of inherited diseases with abnormal N-glycosylation of glycoconjugates. Most patients have CDG-Ia, which is due to a phosphomannomutase (PMM) deficiency. In this article, we report that a significant portion (9 of 54) of patients with CDG-Ia had a rather high residual PMM activity in fibroblasts included in the normal range (means of the controls ± 2 SD) and amounting to 35%–70% of the mean control value. The clinical ...

  16. Activated carbon derived from carbon residue from biomass gasification and its application for dye adsorption: Kinetics, isotherms and thermodynamic studies.

    Science.gov (United States)

    Maneerung, Thawatchai; Liew, Johan; Dai, Yanjun; Kawi, Sibudjing; Chong, Clive; Wang, Chi-Hwa

    2016-01-01

    In this work, activated carbon (AC) as an effective and low-cost adsorbent was successfully prepared from carbon residue (or char, one of the by-products from woody biomass gasification) via physical activation. The surface area of char was significantly increased from 172.24 to 776.46m(2)/g after steam activation at 900°C. The obtained activated carbons were then employed for the adsorption of dye (Rhodamine B) and it was found that activated carbon obtained from steam activation exhibited the highest adsorption capability, which is mainly attributed to the higher surface area and the abundance of hydroxyl (-OH) and carboxyl (-COOH) groups on the activated carbon surface. Moreover, it was also found that the adsorption capability significantly increased under the basic condition, which can be attributed to the increased electrostatic interaction between the deprotonated (negatively charged) activated carbon and dye molecules. Furthermore, the equilibrium data were fitted into different adsorption isotherms and found to fit well with Langmuir model (indicating that dye molecules form monolayer coverage on activated carbon) with a maximum monolayer adsorption capability of 189.83mg/g, whereas the adsorption kinetics followed the pseudo-second-order kinetics. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Biochemical characterization of mutants in the active site residues of the β-galactosidase enzyme of Bacillus circulans ATCC 31382

    Directory of Open Access Journals (Sweden)

    Jelle B. Bultema

    2014-01-01

    Full Text Available The Bacillus circulans ATCC 31382 β-galactosidase (BgaD is a retaining-type glycosidase of glycoside hydrolase family 2 (GH2. Its commercial enzyme preparation, Biolacta N5, is used for commercial-scale production of galacto-oligosaccharides (GOS. The BgaD active site and catalytic amino acid residues have not been studied. Using bioinformatic routines we identified two putative catalytic glutamates and two highly conserved active site histidines. The site-directed mutants E447N, E532Q, and H345F, H379F had lost (almost all catalytic activity. This confirmed their essential role in catalysis, as general acid/base catalyst (E447 and nucleophile (E532, and as transition state stabilizers (H345, H379, respectively.

  18. Conditioning of high activity solid waste: fuel claddings and dissolution residues

    International Nuclear Information System (INIS)

    Anon.

    1982-01-01

    This chapter reports on experimental studies of embedding into matrix material, the melting and conversion of zircaloy, and waste properties and characterization. Methods are developed for embedding the waste scrap into a solid and resistant matrix material in order to confine the radioactivity and to prevent it from dispersion. The matrix materials investigated are lead alloys, ceramics and compacted graphite or aluminium powder. The treatment of fuel hulls by melting or chemical conversion is described. Cemented hulls are characterized and the pyrophoricity of zircaloy fines is investigated. Topics considered include the embedding of hulls into graphite and aluminium, the embedding of hulls and dissolution residues into alumino-ceramics, the solidification of alpha-bearing wastes into a ceramic matrix, and the conditioning of cladding waste by eutectoidic melting and by embedding in glass

  19. Maize, Sunflower and Barley Sensitivity to the Residual Activity of Clomazone in Soil

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    Jelena Gajić Umiljendić

    2012-01-01

    Full Text Available Sensitivity of maize, sunflower and barley to clomazone residues in loamy soil wasassessed in the study using bioassay. Clomazone was applied at a series of concentrationsfrom 0.12 to 12 mg a.i./kg of soil. After 14 days, morphological (shoot height, fresh and dryweight and physiological (content of carotenoids, chlorophyll a and chlorophyll b parameterswere measured. The results showed that morphological parameters are not valid indicatorsof clomazone sensitivity. Based on the results showing inhibition of the physiologicalparameters, I50 values were calculated and used to estimate the difference in sensitivitybetween the species tested. Sunflower was the most sensitive species, while the differencein sensitivity between maize and barley was not significant.Nomenclature: clomazone (2-(2-chlorbenzyl-4,4-dimethyl-1,2-oxazolidin-3-one, maize(Zea mays L., sunflower (Helianthus annuus L., barley (Hordeum vulgare L.

  20. Mutational analysis of an archaeal minichromosome maintenance protein exterior hairpin reveals critical residues for helicase activity and DNA binding

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    Brewster Aaron S

    2010-08-01

    Full Text Available Abstract Background The mini-chromosome maintenance protein (MCM complex is an essential replicative helicase for DNA replication in Archaea and Eukaryotes. While the eukaryotic complex consists of six homologous proteins (MCM2-7, the archaeon Sulfolobus solfataricus has only one MCM protein (ssoMCM, six subunits of which form a homohexamer. We have recently reported a 4.35Å crystal structure of the near full-length ssoMCM. The structure reveals a total of four β-hairpins per subunit, three of which are located within the main channel or side channels of the ssoMCM hexamer model generated based on the symmetry of the N-terminal Methanothermobacter thermautotrophicus (mtMCM structure. The fourth β-hairpin, however, is located on the exterior of the hexamer, near the exit of the putative side channels and next to the ATP binding pocket. Results In order to better understand this hairpin's role in DNA binding and helicase activity, we performed a detailed mutational and biochemical analysis of nine residues on this exterior β-hairpin (EXT-hp. We examined the activities of the mutants related to their helicase function, including hexamerization, ATPase, DNA binding and helicase activities. The assays showed that some of the residues on this EXT-hp play a role for DNA binding as well as for helicase activity. Conclusions These results implicate several current theories regarding helicase activity by this critical hexameric enzyme. As the data suggest that EXT-hp is involved in DNA binding, the results reported here imply that the EXT-hp located near the exterior exit of the side channels may play a role in contacting DNA substrate in a manner that affects DNA unwinding.

  1. Depth profiling of residual activity of ^{237}U fragments as a range verification technique for ^{238}U primary ion beam

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    I. Strašík

    2012-07-01

    Full Text Available Experimental and simulation data concerning fragmentation of ^{238}U ion beam in aluminum, copper, and stainless-steel targets with the initial energy 500 and 950  MeV/u are collected in the paper. A range-verification technique based on depth profiling of residual activity is presented. The irradiated targets were constructed in the stacked-foil geometry and analyzed using gamma-ray spectroscopy. One of the purposes of these experiments was depth profiling of residual activity of induced nuclides and projectile fragments. Among the projectile fragments, special attention is paid to the ^{237}U isotope that has a range very close to the range of the primary ^{238}U ions. Therefore, the depth profiling of the ^{237}U isotope can be utilized for experimental verification of the ^{238}U primary-beam range, which is demonstrated and discussed in the paper. The experimental data are compared with computer simulations by FLUKA, SRIM, and ATIMA, as well as with complementary experiments.

  2. Differential Effects of Continuous Exposure to the Investigational Metastin/Kisspeptin Analog TAK-683 on Pulsatile and Surge Mode Secretion of Luteinizing Hormone in Ovariectomized Goats

    Science.gov (United States)

    TANAKA, Tomomi; OHKURA, Satoshi; WAKABAYASHI, Yoshihiro; KUROIWA, Takenobu; NAGAI, Kiyosuke; ENDO, Natsumi; TANAKA, Akira; MATSUI, Hisanori; KUSAKA, Masami; OKAMURA, Hiroaki

    2013-01-01

    Abstract The aim of the present study was to determine if the estradiol-induced luteinizing hormone (LH) surge is influenced by the constant exposure to TAK-683, an investigational metastin/kisspeptin analog, that had been established to depress the pulsatile gonadotropin-releasing hormone (GnRH) and LH secretion in goats. Ovariectomized goats subcutaneously received TAK-683 (TAK-683 group, n=6) or vehicle (control group, n=6) constantly via subcutaneous implantation of an osmotic pump. Five days after the start of the treatment, estradiol was infused intravenously in both groups to evaluate the effects on the LH surge. Blood samples were collected at 6-min intervals for 4 h prior to the initiation of either the TAK-683 treatment or the estradiol infusion, to determine the profiles of pulsatile LH secretion. They were also collected at 2-h intervals from –4 h to 32 h after the start of estradiol infusion for analysis of LH surges. The frequency and mean concentrations of LH pulses in the TAK-683 group were remarkably suppressed 5 days after the start of TAK-683 treatment compared with those of the control group (P<0.05). On the other hand, a clear LH surge was observed in all animals of both groups. There were no significant differences in the LH concentrations for surge peak and the peak time of the LH surge between the TAK-683 and control groups. These findings suggest that the effects of continuous exposure to kisspeptin or its analog on the mechanism(s) that regulates the pulsatile and surge mode secretion of GnRH/LH are different in goats. PMID:24047956

  3. Insights into the activity change of spore photoproduct lyase induced by mutations at a peripheral glycine residue

    Science.gov (United States)

    Yang, Linlin; Li, Lei

    2017-03-01

    UV radiation triggers the formation of 5-thyminyl-5,6-dihydrothymine, i.e. the spore photoproduct (SP), in the genomic DNA of bacterial endospores. These SPs, if not repaired in time, may lead to genome instability and cell death. SP is mainly repaired by spore photoproduct lyase (SPL) during spore outgrowth via an unprecedented protein-harbored radical transfer pathway that is composed of at least a cysteine and two tyrosine residues. This mechanism is consistent with the recently solved SPL structure that shows all three residues are located in proximity and thus able to participate in the radical transfer process during the enzyme catalysis. In contrast, an earlier in vivo mutational study identified a glycine to arginine mutation at the position 168 on the B. subtilis SPL that was later found to be > 15 Å away from the enzyme active site. This mutation appears to abolish the enzyme activity because endospores carrying this mutant were sensitive to UV light. To understand the molecular basis for this rendered enzyme activity, we constructed two SPL mutations G168A and G168R, examined their repair of dinucleotide SP TpT, and found that both mutants exhibit reduced enzyme activity. Comparing with the wildtype (WT) SPL enzyme, the G168A mutant slows down the SP TpT repair by 3 4 fold while the G168R mutant by 80 fold. Both mutants exhibit a smaller apparent (DV) kinetic isotope effect (KIE) but a bigger competitive (DV/K) KIE than that by the WT SPL. Moreover, the G168R mutant also produces a large portion of the abortive repair product TpT-SO2-; the formation of which indicates that cysteine 141 is no longer well positioned as the H-donor to the thymine allylic radical intermediate. All these data imply that the mutation at the remote glycine 168 residue alters the enzyme 3D structure, subsequently reducing the SPL activity by changing the positions of the essential amino acids involved in the radical transfer process.

  4. Determination of activity by gamma spectrometry of radionuclides present in drums of residues generated in nuclear centrals

    International Nuclear Information System (INIS)

    Aguiar, J.C.; Fernandez, J.

    2006-01-01

    The generation of radioactive residuals in nuclear centrals as CNA I (Atucha I Nuclear Central) and CNE (Embalse Nuclear Central) makes that the measurement of those radionuclides has been a previous stage to the waste management. A method used in those nuclear centrals it is the gamma spectrometry with HPGe detectors, previous to the immobilization of the residual in a cemented matrix, with this the contact with the external agents and its possible dispersion to the atmosphere in the short term is avoided. The ARN (Nuclear Regulatory Authority) of Argentina it carries out periodically intercomparisons and evaluations of the measurement and procedures systems used in the nuclear power stations for the correct measurement and determination of activity of radioactive residuals by gamma spectrometry. In this work an independent method of measurement is exposed to the nuclear power stations. To determine the activity of the residuals by gamma spectrometry deposited in drums, it is required of the precise knowledge of the efficiency curve for such geometry and matrix. Due to the RNA doesn't have a pattern of these characteristics, a mathematical model has been used to obtain this efficiency curve. For it, it is necessary to determine previously: 1) the geometric efficiency or solid angle sustained by the source-detector system (drum-detector) applying a mathematical model described in this work. 2) To estimate the auto-attenuation factor that present the photons in the cemented matrix, these calculations are carried out with a simple equation and its are verified with the Micro Shield 6.10 program. The container commonly used by these nuclear power stations its are drums for 220 liters constructed with SAE 1010 steel and with a thickness of 0.127 cm, with an approximate weight 7.73 Kg., internal diameter of 57.1 cm, and height: 87 cm. The results obtained until the moment register a discrepancy from 5 to 10% with relationship to the measurements carried out by the

  5. Neolithic and Eneolithic activities inferred from organic residue analysis of pottery from Mala Triglavca, Moverna vas and Ajdovska jama, Slovenia

    Directory of Open Access Journals (Sweden)

    Lucija Šoberl

    2014-12-01

    Full Text Available The research discussed in this paper focused on the analysis and identification of organic residues either preserved as visible or absorbed organic remains on Neolithic and Eneolithic pottery from various archaeological and geographical contexts. These are connected with various food preparation strategies and past human activities, i.e. cave burials in Ajdovska jama (food as a grave good/offering, the rock shelter at Mala Triglavca (meat and dairy animal husbandry practices and Moverna vas, which had a long occupation sequence (complex farming and animal management. The preservation of biomarkers mirrored past human activities and different pottery uses at various types of sites. The carbon stable isotope ratios of primary fatty acids in lipid pottery extracts confirmed the presence of adipose and dairy fats as well as biomarkers of plant fats, beeswax and birch bark tar.

  6. RESIDUAL ACTIVITY OF HERBICIDES APPLIED TO COTTON ON CROPS CULTIVATED IN SUCCESSION

    Directory of Open Access Journals (Sweden)

    ELIEZER ANTONIO GHENO

    2016-01-01

    Full Text Available Herbicides with high persistence in soil can cause problems for crops sown in succession to their application. Thus, the aim of this study was to estimate, in greenhouse conditions, the safe period of time after application of preemergent herbicides used on cotton crops (isolated or in mixtures for the crops grown in succession (bean, corn, and soybean. The experimental design was completely randomized in a factorial scheme (5 x 11 + 1, with five repetitions. For each experiment, treatments combined different time periods between herbicide application and sowing of crops (280, 210, 140, 70, and 0 days before sowing of crops with eleven herbicide treatments: fomesafen (625 g ha - 1 prometryne (1250 g ha - 1 , diuron (1250 g ha - 1 , S - metolachlor (768 g ha - 1 , clomazone (1000 g ha - 1 , fomesafen + prometryne (625 + 1250 g ha - 1 , fomesafen + diuron (625 + 1250 g ha - 1 , fomesafen + S - metolachlor (625 + 768 g ha - 1 , fomesafen + clomazone (625 + 1000 g ha - 1 , fomesafen + clomazone + diuron (625 + 1000 + 1250 g ha - 1 , and fomesafen + clomazone + prometryne (625 + 1000 + 1250 g ha - 1 , plus an untreated control. Applications of diuron showed the greatest persistence, causing the largest carryover effects for the three crops evaluated. The other treatments showed residual effects or affected crop development when sowings were performed up to 70 days after application. At later periods no significant damage was observed.

  7. Activation of peroxymonosulfate using drinking water treatment residuals for the degradation of atrazine.

    Science.gov (United States)

    Zhang, Huijuan; Liu, Xitao; Ma, Jun; Lin, Chunye; Qi, Chengdu; Li, Xiaowan; Zhou, Zhou; Fan, Guoxuan

    2018-02-15

    Drinking water treatment residuals (WTRs) are safe byproducts of water treatment plants containing iron. This work studies the degradation of atrazine (ATZ) by WTR-catalyzed peroxymonosulfate (PMS) in aqueous solutions. Factors that affect the catalytic performance (the PMS concentration, catalyst dose, initial solution pH, reaction temperature and water matrix species) were investigated. The results show that the catalytic degradation efficiency of ATZ increases with the increase in PMS concentration and temperature, whereas a higher content of WTRs results in lower removal efficiency because of the quenching effect and negative effect of high pH. For an initial solution pH of 3 and 5, 94.1% and 87.4% of ATZ degradation can be achieved within 6h, whereas the value is only 26% for pH of 7. The production of sulfate radicals (SO 4 - ) and hydroxyl radicals (OH) was confirmed by classic radical quenching and electron spin resonance (ESR) tests. Based on the GC-MS and LC-MS results, the main degradation pathways of ATZ may contain dealkylation, dechlorination-hydroxylation, and alkyl chain oxidation processes. In addition to the ATZ removal ability, the WTRs/PMS system can simultaneously remove phosphorus. This article provides a new idea for wastewater treatment and usage of WTRs as a resource. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Activated carbons from waste biomass: an alternative use for biodiesel production solid residues.

    Science.gov (United States)

    Nunes, Anne A; Franca, Adriana S; Oliveira, Leandro S

    2009-03-01

    Defective coffee press cake, a residue from coffee oil biodiesel production, was evaluated as raw material for production of an adsorbent for removal of methylene blue (MB) from aqueous solution. Batch adsorption tests were performed at 25 degrees C and the effects of particle size, contact time, adsorbent dosage and pH were investigated. Preliminary adsorption tests indicated that thermal treatment is necessary in order to improve adsorption capacity. Adsorption kinetics was determined by fitting first and second-order kinetic models to the experimental data, with the second-order model providing the best description of MB adsorption onto the prepared adsorbent. The experimental adsorption equilibrium data were fitted to Langmuir, Freundlich and Temkin adsorption models, with the last two providing the best fits. The experimental data obtained in the present study indicated that this type of waste material is a suitable candidate for use in the production of adsorbents for removal of cationic dyes, thus contributing for the implementation of sustainable development in both the coffee and biodiesel production chains.

  9. GW501516, a PPARδ agonist, ameliorates tubulointerstitial inflammation in proteinuric kidney disease via inhibition of TAK1-NFκB pathway in mice.

    Directory of Open Access Journals (Sweden)

    Xu Yang

    Full Text Available Peroxisome proliferator-activated receptors (PPARs are a nuclear receptor family of ligand-inducible transcription factors, which have three different isoforms: PPARα, δ and γ. It has been demonstrated that PPARα and γ agonists have renoprotective effects in proteinuric kidney diseases; however, the role of PPARδ agonists in kidney diseases remains unclear. Thus, we examined the renoprotective effect of GW501516, a PPARδ agonist, in a protein-overload mouse nephropathy model and identified its molecular mechanism. Mice fed with a control diet or GW501516-containing diet were intraperitoneally injected with free fatty acid (FFA-bound albumin or PBS(-. In the control group, protein overload caused tubular damages, macrophage infiltration and increased mRNA expression of MCP-1 and TNFα. These effects were prevented by GW501516 treatment. In proteinuric kidney diseases, excess exposure of proximal tubular cells to albumin, FFA bound to albumin or cytokines such as TNFα is detrimental. In vitro studies using cultured proximal tubular cells showed that GW501516 attenuated both TNFα- and FFA (palmitate-induced, but not albumin-induced, MCP-1 expression via direct inhibition of the TGF-β activated kinase 1 (TAK1-NFκB pathway, a common downstream signaling pathway to TNFα receptor and toll-like receptor-4. In conclusion, we demonstrate that GW501516 has an anti-inflammatory effect in renal tubular cells and may serve as a therapeutic candidate to attenuate tubulointerstitial lesions in proteinuric kidney diseases.

  10. Active subsite properties, subsite residues and targeting to lysosomes or midgut lumen of cathepsins L from the beetle Tenebrio molitor.

    Science.gov (United States)

    Damasceno, Ticiane F; Dias, Renata O; de Oliveira, Juliana R; Salinas, Roberto K; Juliano, Maria A; Ferreira, Clelia; Terra, Walter R

    2017-10-01

    Cathepsins L are the major digestive peptidases in the beetle Tenebrio molitor. Two digestive cathepsins L (TmCAL2 and TmCAL3) from it had their 3D structures solved. The aim of this paper was to study in details TmCAL3 specificity and properties and relate them to its 3D structure. Recombinant TmCAL3 was assayed with 64 oligopeptides with different amino acid replacements in positions P2, P1, P1' and P2'. Results showed that TmCAL3 S2 specificity differs from the human enzyme and that its specificities also explain why on autoactivation two propeptide residues remain in the enzyme. Data on free energy of binding and of activation showed that S1 and S2' are mainly involved in substrate binding, S1' acts in substrate binding and catalysis, whereas S2 is implied mainly in catalysis. Enzyme subsite residues were identified by docking with the same oligopeptide used for kinetics. The subsite hydrophobicities were calculated from the efficiency of hydrolysis of different amino acid replacements in the peptide and from docking data. The results were closer for S1 and S2' than for S1' and S2, indicating that the residue subsites that were more involved in transition state binding are different from those binding the substrate seen in docking. Besides TmCAL1-3, there are nine other cathepsins L, most of them more expressed at midgut. They are supposed to be directed to lysosomes by a Drosophila-like Lerp receptor and/or motifs in their prodomains. The mannose 6-phosphate lysosomal sorting machinery is absent from T. molitor transcriptome. Cathepsin L direction to midgut contents seems to depend on overexpression. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Determination of platinum group metal catalyst residues in active pharmaceutical ingredients by means of total reflection X-ray spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Marguí, Eva, E-mail: eva.margui@udg.edu [Department of Chemistry, University of Girona, Campus Montilivi, 17071 Girona (Spain); Queralt, Ignasi, E-mail: iqueralt@ictja.csic.es [Laboratory of X-ray Analytical Applications, Institute of Earth Sciences “Jaume Almera”, CSIC, Solé Sabarís s/n, 08028 Barcelona (Spain); Hidalgo, Manuela, E-mail: manuela.hidalgo@udg.edu [Department of Chemistry, University of Girona, Campus Montilivi, 17071 Girona (Spain)

    2013-08-01

    The control of metal catalyst residues (i.e., platinum group metals (PGMs)) in different stages of the manufacturing processes of the active pharmaceutical ingredients (APIs) and, especially, in the final product is crucial. For API specimens, there are strict guidelines to limit the levels of metal residues based on their individual levels of safety concern. For PGMs the concentration limit has been established at 10 mg/kg in the API. Therefore great effort is currently being devoted to the development of new and simple procedures to control metals in pharmaceuticals. In the present work, an analytical methodology based on benchtop total reflection X-ray fluorescence spectrometry (TXRF) has been developed for the rapid and simple determination of some PGM catalyst impurities (Rh, Pd, Ir and Pt) in different types of API samples. An evaluation of different sample treatments (dissolution and digestion of the solid pharmaceutical samples) has been carried out and the developed methodologies have been validated according to the analytical parameters to be considered and acceptance criteria for PGM determination according to the United States Pharmacopeia (USP). Limits of quantification obtained for PGM metals were in the range of 2–4 mg/kg which are satisfactory according to current legislation. From the obtained results it is shown that the developed TXRF method can be implemented in the pharmaceutical industries to increase productivity of the laboratory; offering an interesting and complementary analytical tool to other atomic spectroscopic methods. - Highlights: • A TXRF method for PGM catalyst residue determination in API samples is presented. • Analysis can be performed using 10 μL of the internal standardized dissolved API. • The method is rapid, simple and suitable according to the USP requirements.

  12. H-bonding networks of the distal residues and water molecules in the active site of Thermobifida fusca hemoglobin.

    Science.gov (United States)

    Nicoletti, Francesco P; Droghetti, Enrica; Howes, Barry D; Bustamante, Juan P; Bonamore, Alessandra; Sciamanna, Natascia; Estrin, Darío A; Feis, Alessandro; Boffi, Alberto; Smulevich, Giulietta

    2013-09-01

    The ferric form of truncated hemoglobin II from Thermobifida fusca (Tf-trHb) and its triple mutant WG8F-YB10F-YCD1F at neutral and alkaline pH, and in the presence of CN(-) have been characterized by resonance Raman spectroscopy, electron paramagnetic resonance spectroscopy, and molecular dynamics simulations. Tf-trHb contains three polar residues in the distal site, namely TrpG8, TyrCD1 and TyrB10. Whereas TrpG8 can act as a potential hydrogen-bond donor, the tyrosines can act as donors or acceptors. Ligand binding in heme-containing proteins is determined by a number of factors, including the nature and conformation of the distal residues and their capability to stabilize the heme-bound ligand via hydrogen-bonding and electrostatic interactions. Since both the RR Fe-OH(-) and Fe-CN(-) frequencies are very sensitive to the distal environment, detailed information on structural variations has been obtained. The hydroxyl ligand binds only the WT protein giving rise to two different conformers. In form 1 the anion is stabilized by H-bonds with TrpG8, TyrCD1 and a water molecule, in turn H-bonded to TyrB10. In form 2, H-bonding with TyrCD1 is mediated by a water molecule. Unlike the OH(-) ligand, CN(-) binds both WT and the triple mutant giving rise to two forms with similar spectroscopic characteristics. The overall results clearly indicate that H-bonding interactions both with distal residues and water molecules are important structural determinants in the active site of Tf-trHb. This article is part of a Special Issue entitled: Oxygen Binding and Sensing Proteins. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. How to reduce injuries to residual trees during stand management activities.

    Science.gov (United States)

    Paul E. Aho; Gary Fiddler; Gregory M. Filip

    1983-01-01

    Losses of trees and tree volume that result from decay initiated by mechanical injuries during stand management activities in the western United States are substantial. They can be reduced through improved logging methods and careful planning of other forest management activities.

  14. On the catalytic role of the active site residue E121 of E. coli L-aspartate oxidase.

    Science.gov (United States)

    Tedeschi, Gabriella; Nonnis, Simona; Strumbo, Bice; Cruciani, Gabriele; Carosati, Emanuele; Negri, Armando

    2010-10-01

    L-aspartate oxidase (LASPO) is a flavoenzyme catalyzing the first step in the de novo biosynthesis of NAD+. The enzyme oxidizes L-aspartate both under aerobic and anaerobic conditions using oxygen as well as fumarate as electron acceptor. In accordance with its catalytic activities, LASPO displays strong primary and tertiary structure similarity with the flavin containing subunit of the proteins belonging to the succinate dehydrogenase/fumarate reductase family. The similarity extends to the active site residues, with LASPO differing from the other enzymes of the family only for the presence of a conserved glutamate (E121), which is substituted by apolar amino acids in the other enzymes. Three complementary approaches have been used to define the role of E121 in LASPO: characterization of mutants (E121A, E121Q, E121D and E121K), investigation of the catalytic activities of WT and mutants towards substrates and substrate analogues and molecular docking studies. All mutants retain fumarate reductase activity. On the contrary, all mutants lack L-aspartate oxidase activity, although retaining the ability to bind L-aspartate (except for E121K). These results and investigations on the oxidase activity towards substrate analogues suggest that the roles of E121 in catalysis include orienting L-aspartate in a productive binding mode and favouring proton abstraction from C2 by an active site base. Molecular docking studies of the substrate (L-aspartate), inhibitor (D-aspartate) and product (imino aspartate) in the active site of LASPO confirm that (a) the substrate/product energetically favoured orientation in the active site supports the conclusions reported above, (b) E121 interacts favourably with the charged amino group of the substrate and (c) different ligands might assume different orientations in the active site of the enzyme. Copyright © 2010 Elsevier Masson SAS. All rights reserved.

  15. Bioactive Properties of Phaseolus lunatus (Lima Bean) and Vigna unguiculata (Cowpea) Hydrolyzates Incorporated into Pasta. Residual Activity after Pasta Cooking.

    Science.gov (United States)

    Drago, Silvina R; Franco-Miranda, Hanai; Cian, Raúl E; Betancur-Ancona, David; Chel-Guerrero, Luis

    2016-09-01

    The aims of the study were to study the inclusion of P. lunatus (PLH) and V. unguiculata (VUH) protein hydrolyzates with bioactive properties into a pasta-extruded product and determine residual activity after extrusion or pasta cooking. Both protein hydrolyzates showed angiotensin-converting enzyme inhibition (ACEI) and antioxidant activity (TEAC). PLH showed higher ACEI but lower TEAC than VUH (97.19 ± 0.23 vs. 91.95 ± 0.29 % and 244.7 ± 3.4 vs. 293.7 ± 3.3 μmol Trolox/g, respectively). They were included at 5 or 10 % into wheat pasta. Control pasta had the lowest ACEI activity or TEAC (22.01 ± 0.76 % or 14.14 ± 1.28 μmol Trolox/g, respectively). Higher activity remained in pasta with PLH than VUH after extrusion, and higher the level of addition, higher the ACEI was. Pasta had practically the same ACEI activity after cooking, thus active compounds were not lost by temperature or lixiviation. Regarding TEAC, higher activity remained in pasta with 10 % VUH (31.84 ± 0.17 μmol Trolox/g). Other samples with hydrolyzates had the same activity. After cooking, pasta with hydrolyzates had higher TEAC values than control, but these were not modified by the level of incorporation. Moreover, the profile changed because pasta with PLH had the highest TEAC values (21.39 ± 0.01 and 20.34 ± 0.15 for 5 or 10 % hydrolyzates, respectively). Cooking decreased this activity (~ 20 %), for all samples. Although a certain loss of antioxidant activity was observed, pasta could be a good vehicle for bioactive compounds becoming a functional food.

  16. Redox-active tyrosine residue in the microcin J25 molecule

    Energy Technology Data Exchange (ETDEWEB)

    Chalon, Miriam C. [Departamento de Bioquimica de la Nutricion, Instituto Superior de Investigaciones Biologicas (Consejo Nacional de Investigaciones Cientificas y Tecnicas-Universidad Nacional de Tucuman) and Instituto de Quimica Biologica ' Dr Bernabe Bloj' , Chacabuco 461, 4000 San Miguel de Tucuman, Tucuman (Argentina); Wilke, Natalia [CIQUIBIC, Dpto. de Quimica Biologica, Facultad de Ciencias Quimicas, Universidad Nacional de Cordoba, Pabellon Argentina, Ciudad Universitaria, X5000HUA, Cordoba (Argentina); Pedersen, Jens; Rufini, Stefano [Department of Biology, University of Rome Tor Vergata 00133, Rome (Italy); Morero, Roberto D.; Cortez, Leonardo; Chehin, Rosana N.; Farias, Ricardo N. [Departamento de Bioquimica de la Nutricion, Instituto Superior de Investigaciones Biologicas (Consejo Nacional de Investigaciones Cientificas y Tecnicas-Universidad Nacional de Tucuman) and Instituto de Quimica Biologica ' Dr Bernabe Bloj' , Chacabuco 461, 4000 San Miguel de Tucuman, Tucuman (Argentina); Vincent, Paula A., E-mail: pvincent@fbqf.unt.edu.ar [Departamento de Bioquimica de la Nutricion, Instituto Superior de Investigaciones Biologicas (Consejo Nacional de Investigaciones Cientificas y Tecnicas-Universidad Nacional de Tucuman) and Instituto de Quimica Biologica ' Dr Bernabe Bloj' , Chacabuco 461, 4000 San Miguel de Tucuman, Tucuman (Argentina)

    2011-03-18

    Research highlights: {yields} Cyclic voltammetry measurements showed irreversible oxidation of MccJ25 and MccJ25 (Y9F). {yields} Infrared spectroscopy studies showed that only Tyr9 could be deprotonated upon chemical oxidation. {yields} Formation of a long-lived tyrosyl radical in the native MccJ25 oxidized by H{sub 2}O{sub 2} was demonstrated. {yields} Tyr9 but not Tyr20 can be easily oxidized and form a tyrosyl radical. -- Abstract: Microcin J25 (MccJ25) is a 21 amino acid lasso-peptide antibiotic produced by Escherichia coli and composed of an 8-residues ring and a terminal 'tail' passing through the ring. We have previously reported two cellular targets for this antibiotic, bacterial RNA polymerase and the membrane respiratory chain, and shown that Tyr9 is essential for the effect on the membrane respiratory chain which leads to superoxide overproduction. In the present paper we investigated the redox behavior of MccJ25 and the mutant MccJ25 (Y9F). Cyclic voltammetry measurements showed irreversible oxidation of both Tyr9 and Tyr20 in MccJ25, but infrared spectroscopy studies demonstrated that only Tyr9 could be deprotonated upon chemical oxidation in solution. Formation of a long-lived tyrosyl radical in the native MccJ25 oxidized by H{sub 2}O{sub 2} was demonstrated by Electron Paramagnetic Resonance Spectroscopy; this radical was not detected when the reaction was carried out with the MccJ25 (Y9F) mutant. These results show that the essential Tyr9, but not Tyr20, can be easily oxidized and form a tyrosyl radical.

  17. Redox-active tyrosine residue in the microcin J25 molecule

    International Nuclear Information System (INIS)

    Chalon, Miriam C.; Wilke, Natalia; Pedersen, Jens; Rufini, Stefano; Morero, Roberto D.; Cortez, Leonardo; Chehin, Rosana N.; Farias, Ricardo N.; Vincent, Paula A.

    2011-01-01

    Research highlights: → Cyclic voltammetry measurements showed irreversible oxidation of MccJ25 and MccJ25 (Y9F). → Infrared spectroscopy studies showed that only Tyr9 could be deprotonated upon chemical oxidation. → Formation of a long-lived tyrosyl radical in the native MccJ25 oxidized by H 2 O 2 was demonstrated. → Tyr9 but not Tyr20 can be easily oxidized and form a tyrosyl radical. -- Abstract: Microcin J25 (MccJ25) is a 21 amino acid lasso-peptide antibiotic produced by Escherichia coli and composed of an 8-residues ring and a terminal 'tail' passing through the ring. We have previously reported two cellular targets for this antibiotic, bacterial RNA polymerase and the membrane respiratory chain, and shown that Tyr9 is essential for the effect on the membrane respiratory chain which leads to superoxide overproduction. In the present paper we investigated the redox behavior of MccJ25 and the mutant MccJ25 (Y9F). Cyclic voltammetry measurements showed irreversible oxidation of both Tyr9 and Tyr20 in MccJ25, but infrared spectroscopy studies demonstrated that only Tyr9 could be deprotonated upon chemical oxidation in solution. Formation of a long-lived tyrosyl radical in the native MccJ25 oxidized by H 2 O 2 was demonstrated by Electron Paramagnetic Resonance Spectroscopy; this radical was not detected when the reaction was carried out with the MccJ25 (Y9F) mutant. These results show that the essential Tyr9, but not Tyr20, can be easily oxidized and form a tyrosyl radical.

  18. ADAMTS13 Secretion and Residual Activity among Patients with Congenital Thrombotic Thrombocytopenic Purpura with and without Renal Impairment

    Science.gov (United States)

    Rurali, Erica; Banterla, Federica; Donadelli, Roberta; Bresin, Elena; Galbusera, Miriam; Gastoldi, Sara; Peyvandi, Flora; Underwood, Mary; Noris, Marina

    2015-01-01

    Background and objectives Acute renal impairment is observed in 11%–23% of patients with congenital thrombotic thrombocytopenic purpura (TTP) and deficiency of a disintegrin and metalloprotease with thrombospondin motifs 13 (ADAMTS13, a metalloprotease that cleaves von Willebrand factor [VWF] multimers), a substantial percentage of whom develop CKD during follow-up. Design, setting, participants, & measurements Here we investigated whether, in 18 patients with congenital recruited from 1996 to 2013 who fulfilled inclusion criteria, acute renal involvement occurred during bouts segregated with lower secretion and activity levels of ADAMTS13 mutants. We performed expression studies and a sensitive recombinant VWF (rVWF) A1-A2-A3 cleavage test (detection limit, 0.78% of normal ADAMTS13 activity). Results A higher risk of acute renal impairment during bouts was observed in patients with childhood (<18 years) onset (odds ratio [OR], 24.6 [95% confidence interval (CI), 1.11 to 542.44]) or a relapsing (≥1 episode per year) disease (OR, 54.6 [95% CI, 2.25 to 1326.28]) than in patients with adulthood onset or long-lasting remission, respectively. Whatever the age at onset, patients with acute renal impairment had mutations different from those in patients without renal involvement. Moreover, mutations in patients with acute renal impairment compared with those in patients without renal involvement caused lower in vitro rADAMTS13 secretion (1.33% versus 12.5%; P<0.001) and residual activity (0.11% versus 3.47%; P=0.003). rADAMTS13 secretion ≤3.75% and residual activity ≤0.4% best discriminated patients with renal impairment (receiver-operating characteristic curve sensitivity, 100% and 100%; specificity, 100% and 83.3%, respectively; logistic regression OR, 325 [95% CI, 6 to 18339] and 91.7 [95% CI, 3.2 to 2623.5], respectively). All mutations found in patients with childhood onset or relapsing disease were associated with acute renal impairment during bouts

  19. Preparation, characterization, and α-glycosidase inhibition activity of a carboxymethylated polysaccharide from the residue of Sarcandra glabra (Thunb.) Nakai.

    Science.gov (United States)

    Liu, Wei; Hu, Cheng; Liu, Yameng; Dai, Shijie; Lu, Weisheng; Lv, Xing; Yao, Wenbing; Gao, Xiangdong

    2017-06-01

    A carboxymethylated polysaccharide (CMSERP) was prepared from the residue of Sarcandra glabra (Thunb.) Nakai. CMSERP was mainly composed of galacturonic acid (GalA), glucose (Glc), galactose (Gal), glucuronic acid (GlcA), arabinose (Ara), rhamnose (Rha), xylose (Xyl), ribose (Rib), and fucose (Fuc) at the ratio of 29.79:19.30:11.92:6.32:4.68:3.95:3.39:2.31:1.00. The primary structure features of CMSERP were determined to be a pectin like polysaccharide according to FT-IR, NMR, and HPAEC-PAD. The results of HPSEC-MALLS-RID and DLS indicated the Mw, Mn, Mz, and S2Z1/2 of CMSERP were 5.515×10 4 g/mol, 1.566×10 4 g/mol, 1.510×10 6 g/mol, and 62.8 (±1.2%) nm, respectively. TEM and AFM revealed CMSERP was dispersed in 0.05M sodium sulfate but aggregated in water. Moreover, a high α-glucosidase inhibition activity (83.38%±2.30% at 1000μg/mL) of CMSERP which is higher than that of acarbose was observed. The results proved the effects of carboxymethylation on poor water-soluble polysaccharides and explore a potential α-glucosidase inhibitor which from abandoned extracted residue for the functional foods and pharmaceutical industries. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Determination of platinum group metal catalyst residues in active pharmaceutical ingredients by means of total reflection X-ray spectrometry

    Science.gov (United States)

    Marguí, Eva; Queralt, Ignasi; Hidalgo, Manuela

    2013-08-01

    The control of metal catalyst residues (i.e., platinum group metals (PGMs)) in different stages of the manufacturing processes of the active pharmaceutical ingredients (APIs) and, especially, in the final product is crucial. For API specimens, there are strict guidelines to limit the levels of metal residues based on their individual levels of safety concern. For PGMs the concentration limit has been established at 10 mg/kg in the API. Therefore great effort is currently being devoted to the development of new and simple procedures to control metals in pharmaceuticals. In the present work, an analytical methodology based on benchtop total reflection X-ray fluorescence spectrometry (TXRF) has been developed for the rapid and simple determination of some PGM catalyst impurities (Rh, Pd, Ir and Pt) in different types of API samples. An evaluation of different sample treatments (dissolution and digestion of the solid pharmaceutical samples) has been carried out and the developed methodologies have been validated according to the analytical parameters to be considered and acceptance criteria for PGM determination according to the United States Pharmacopeia (USP). Limits of quantification obtained for PGM metals were in the range of 2-4 mg/kg which are satisfactory according to current legislation. From the obtained results it is shown that the developed TXRF method can be implemented in the pharmaceutical industries to increase productivity of the laboratory; offering an interesting and complementary analytical tool to other atomic spectroscopic methods.

  1. In vitro residual anti-bacterial activity of difloxacin, sarafloxacin and their photoproducts after photolysis in water

    International Nuclear Information System (INIS)

    Kusari, Souvik; Prabhakaran, Deivasigamani; Lamshoeft, Marc; Spiteller, Michael

    2009-01-01

    Fluoroquinolones like difloxacin (DIF) and sarafloxacin (SARA) are adsorbed in soil and enter the aquatic environment wherein they are subjected to photolytic degradation. To evaluate the fate of DIF and SARA, their photolysis was performed in water under stimulated natural sunlight conditions. DIF primarily degrades to SARA. On prolonged photodegradation, seven photoproducts were elucidated by HR-LC-MS/MS, three of which were entirely novel. The residual anti-bacterial activities of DIF, SARA and their photoproducts were studied against a group of pathogenic strains. DIF and SARA revealed potency against both Gram-positive and -negative bacteria. The photoproducts also exhibited varying degrees of efficacies against the tested bacteria. Even without isolating the individual photoproducts, their impact on the aquatic environment could be assessed. Therefore, the present results call for prudence in estimating the fate of these compounds in water and in avoiding emergence of resistance in bacteria caused by the photoproducts of DIF and SARA. - Assessment of the residual anti-bacterial efficacies of difloxacin, sarafloxacin and their photoproducts in water, and estimating their impact on the aquatic environment in inducing resistance to microorganisms.

  2. Life stage toxicity and residual activity of insecticides to codling moth and oriental fruit moth (Lepidoptera: Tortricidae).

    Science.gov (United States)

    Magalhaes, Leonardo C; Walgenbach, James F

    2011-12-01

    The codling moth, Cydia pomonella (L.), and oriental fruit moth, Grapholita molesta (Busck), are two key pests of apple (Malus domestica Borkh.) in North Carolina. Growers extensively relied on organophosphate insecticides, primarily azinphosmethyl, for > 40 yr to manage these pests. Because of organophosphate resistance development and regulatory actions, growers are transitioning to management programs that use new, reduced-risk, and OP-replacement insecticides. This study evaluated the toxicity of a diversity of replacement insecticides to eggs, larvae, and adults, as well as an assessment of their residual activity, to codling moth and oriental fruit moth. Laboratory-susceptible strains of both species were used for all bioassays. Fresh field-harvested apples were used as a media for assessing the ovicidal activity of insecticides. For larval studies, insecticides were topically applied to the surface of lima bean-based diet, onto which neonates were placed. Toxicity was based on two measures of mortality; 5-d mortality and development to adult stage. Ovicidal bioassays showed that oriental fruit moth eggs were generally more tolerant than codling moth eggs to insecticides, with novaluron, acetamiprid, and azinphoshmethyl having the highest levels of toxicity to eggs of both species. In contrast, codling moth larvae generally were more tolerant than oriental fruit moth to most insecticides. Methoxyfenozide and pyriproxyfen were the only insecticides with lower LC50 values against codling moth than oriental fruit moth neonates. Moreover, a number of insecticides, particularly the IGRs methoxyfenozide and novaluron, the anthranilic diamide chlorantriliprole, and the spinosyn spinetoram, provided equal or longer residual activity against codling moth compared with azinphosmethyl in field studies. Results are discussed in relation to their use in devising field use patterns of insecticides and for insecticide resistance monitoring programs.

  3. Diagnosis of Compound Heterozygous Hb Tak/β-Thalassemia and HbD-Punjab/β-Thalassemia by HbA2 Levels on Capillary Electrophoresis.

    Science.gov (United States)

    Panyasai, Sitthichai; Sakkhachornphop, Supachai; Pornprasert, Sakorn

    2018-01-01

    A misdiagnosis of β-thalassemia carrier in samples with Hb Tak and HbD-Punjab, the β-variants, can be a cause of inappropriate genetic counseling thus having a new case of β-thalassemia major. A capillary electrophoresis (CE) is very efficient in separating and quantifying HbA 2 . In this study, HbA 2 levels of samples which were doubted for compound heterozygous Hb Tak/β-thalassemia or heterozygous HbD-Punjab/β-thalassemia were measured and compared between CE and high performance liquid chromatography (HPLC). The molecular confirmation for Hb Tak, HbD-Punjab and β-thalassemia codons 17 (A > T), 41/42 (-TCTT), 71/72 (+A) and IVSI-nt1 (G > T) mutations and 3.4 kb deletion were also performed. Based on DNA analysis, 3 cases were diagnosed as compound heterozygous Hb Tak/β-thalassemia and one for HbD-Punjab/β-thalassemia. The elevated HbA 2 levels were found in all 4 samples with rages of 4.6-7.3% on CE while those were not found on HPLC. Thus, the elevated HbA 2 measured by CE can be used as a screening parameter for differentiating the homozygote of Hb Tak and HbD-Punjab from the compound heterozygote of these hemoglobinopathies and β-thalassemia.

  4. Asn12 and Asn278: Critical Residues for In Vitro Biological Activity of Reteplase

    Directory of Open Access Journals (Sweden)

    Naganath Mandi

    2010-01-01

    Full Text Available Reteplase (rPA is a thrombolytic agent used for the treatment of acute myocardial infarction. We studied the expression of rPA and its selected asparagine mutants after integration into the Pichia genome. Though methanol induction of the native and the rPA mutants showed similar expression levels (~200–250 mg/L, the mutants displayed significant loss of protease activity. Strikingly, the clot lysis activities of these mutants were considerably different. While mutation of Asn12 (N12P of the Kringle 2 domain showed delayed clot lysis activity (1/2=38 min compared to the native rPA (1/2=33 min, a faster rate of clot lysis (1/2=27 min was observed when the Asn278 (N278S of the serine protease domain was mutated. Interestingly, the slowest clot lysis activity (1/2=49 min demonstrated by the double mutant (N12P, N278S suggests the dominant role of Asn12 in regulating the fibrinolytic activity of rPA. The results presented in this paper indicate that the fibrinolytic and the proteolytic activities of rPA are independent of each other.

  5. S-nitrosoglutathione covalently modifies cysteine residues of human carbonyl reductase 1 and affects its activity.

    Science.gov (United States)

    Hartmanová, Tereza; Tambor, Vojtěch; Lenčo, Juraj; Staab-Weijnitz, Claudia A; Maser, Edmund; Wsól, Vladimír

    2013-02-25

    Carbonyl reductase 1 (CBR1 or SDR21C1) is a ubiquitously-expressed, cytosolic, monomeric, and NADPH-dependent enzyme. CBR1 participates in apoptosis, carcinogenesis and drug resistance, and has a protective role in oxidative stress, cancer and neurodegeneration. S-Nitrosoglutathione (GSNO) represents the newest addition to its diverse substrate spectrum, which includes a wide range of xenobiotics and endogenous substances. GSNO has also been shown to covalently modify and inhibit CBR1. The aim of the present study was to quantify and characterize the resulting modifications. Of five candidate cysteines for modification by 2 mM GSNO (positions 26, 122, 150, 226, 227), the last four were analyzed using MALDI-TOF/TOF mass spectrometry and then quantified using the Selected Reaction Monitoring Approach on hyphenated HPLC with a triple quadrupole mass spectrometer. The analysis confirmed GSNO concentration-dependent S-glutathionylation of cysteines at positions 122, 150, 226, 227 which was 2-700 times higher compared to wild-type CBR1 (WT-CBR1). Moreover, a disulfide bond between neighboring Cys-226 and Cys-227 was detected. We suggest a role of these two cysteines as a redox-sensitive cysteine pair. The catalytic properties of wild-type and enzyme modified with 2 mM GSNO were also investigated by steady state kinetic experiments with various substrates. GSNO treatment of CBR1 resulted in a 2-5-fold decrease in kcat with menadione, 4-benzoylpyridine, 2,3-hexanedione, daunorubicin and 1,4-naphthoquinone. In contrast, the same treatment increased kcat for substrates containing a 1,2-diketo group in a ring structure (1,2-naphthoquinone, 9,10-phenanthrenequinone, isatin). Except for 9,10-phenanthrenequinone, all changes in kcat were at least in part compensated for by a similar change in Km, overall yielding no drastic changes in catalytic efficiency. The findings indicate that GSNO-induced covalent modification of cysteine residues affects the kinetic mechanism of CBR1

  6. Computation Of The Residual Radionuclide Activity Within Three Natural Waterways At The Savannah River Site

    Energy Technology Data Exchange (ETDEWEB)

    Hiergesell, R. A.; Phifer, M. A.

    2014-01-07

    In 2010 a Composite Analysis (CA) of the U.S. Department of Energy’s (DOE’s) Savannah River Site (SRS) was completed. This investigation evaluated the dose impact of the anticipated SRS End State residual sources of radionuclides to offsite members of the public. Doses were assessed at the locations where SRS site streams discharge into the Savannah River at the perimeter of the SRS. Although the model developed to perform this computation indicated that the dose constraint of 0.3 mSv/yr (30 mrem/yr), associated with CA, was not approached at the Points of Assessment (POAs), a significant contribution to the total computed dose was derived from the radionuclides (primarily Cs-137) bound-up in the soil and sediment of the drainage corridors of several SRS streams. DOE’s Low Level Waste Federal Review Group (LFRG) reviewed the 2010 CA and identified several items to be addressed in the SRS Maintenance Program. One of the items recognized Cs-137 in the Lower Three Runs (LTR) Integrator Operable Unit (IOU), as a significant CA dose driver. The item made the recommendation that SRS update the estimated radionuclide inventory, including Cs-137, in the LTR IOU. That initial work has been completed and its radionuclide inventory refined. There are five additional streams at SRS and the next phase of the response to the LFRG concern was to obtain a more accurate inventory and distribution of radionuclides in three of those streams, Fourmile Branch (FMB), Pen Branch (PB) and Steel Creek (SC). Each of these streams is designated as an IOU, which are defined for the purpose of this investigation as the surface water bodies and associated wetlands, including the channel sediment, floodplain sed/soil, and related biota. If present, radionuclides associated with IOUs are adsorbed to the streambed sediment and soils of the shallow floodplains that lie immediately adjacent to stream channels. The scope of this effort included the evaluation of any previous sampling and

  7. Role of a conserved glutamine residue in tuning the catalytic activity of Escherichia coli cytochrome c nitrite reductase.

    Science.gov (United States)

    Clarke, Thomas A; Kemp, Gemma L; Van Wonderen, Jessica H; Doyle, Rose-Marie A S; Cole, Jeffrey A; Tovell, Nick; Cheesman, Myles R; Butt, Julea N; Richardson, David J; Hemmings, Andrew M

    2008-03-25

    The pentaheme cytochrome c nitrite reductase (NrfA) of Escherichia coli is responsible for nitrite reduction during anaerobic respiration when nitrate is scarce. The NrfA active site consists of a hexacoordinate high-spin heme with a lysine ligand on the proximal side and water/hydroxide or substrate on the distal side. There are four further highly conserved active site residues including a glutamine (Q263) positioned 8 A from the heme iron for which the side chain, unusually, coordinates a conserved, essential calcium ion. Mutation of this glutamine to the more usual calcium ligand, glutamate, results in an increase in the K m for nitrite by around 10-fold, while V max is unaltered. Protein film voltammetry showed that lower potentials were required to detect activity from NrfA Q263E when compared with native enzyme, consistent with the introduction of a negative charge into the vicinity of the active site heme. EPR and MCD spectroscopic studies revealed the high spin state of the active site to be preserved, indicating that a water/hydroxide molecule is still coordinated to the heme in the resting state of the enzyme. Comparison of the X-ray crystal structures of the as-prepared, oxidized native and mutant enzymes showed an increased bond distance between the active site heme Fe(III) iron and the distal ligand in the latter as well as changes to the structure and mobility of the active site water molecule network. These results suggest that an important function of the unusual Q263-calcium ion pair is to increase substrate affinity through its role in supporting a network of hydrogen bonded water molecules stabilizing the active site heme distal ligand.

  8. Role of AP-endonuclease (Ape1) active site residues in stabilization of the reactant enzyme-DNA complex.

    Science.gov (United States)

    Batebi, Hossein; Dragelj, Jovan; Imhof, Petra

    2018-04-01

    Apurinic/apyrimidinic endonuclease 1 (Ape1) is an important metal-dependent enzyme in the base excision repair mechanism, responsible for the backbone cleavage of abasic DNA through a phosphate hydrolysis reaction. Molecular dynamics simulations of Ape1 complexed to its substrate DNA performed for models containing 1 or 2 Mg 2+ -ions as cofactor located at different positions show a complex with 1 metal ion bound on the leaving group site of the scissile phosphate to be the most likely reaction-competent conformation. Active-site residue His309 is found to be protonated based on pKa calculations and the higher conformational stability of the Ape1-DNA substrate complex compared to scenarios with neutral His309. Simulations of the D210N mutant further support the prevalence of protonated His309 and strongly suggest Asp210 as the general base for proton acceptance by a nucleophilic water molecule. © 2018 Wiley Periodicals, Inc.

  9. Comparative estrogenic activity of wine extracts and organochlorine pesticide residues in food

    NARCIS (Netherlands)

    Gaido, K.; Dohme, L.; Wang, F.; Chen, I.; Blankvoort, B.; Ramamoorthy, K.; Safe, S.

    1998-01-01

    The human diet contains industrial-derived, endocrine-active chemicals and higher levels of naturally occurring compounds that modulate multiple endocrine pathways. Hazard and risk assessment of these mixtures is complicated by noadditive interactions between different endocrine-mediated responses.

  10. Toxicity and Residual Activity of Insecticides Against Tamarixia triozae (Hymenoptera: Eulophidae), a Parasitoid of Bactericera cockerelli (Hemiptera: Triozidae).

    Science.gov (United States)

    Luna-Cruz, Alfonso; Rodríguez-Leyva, Esteban; Lomeli-Flores, J Refugio; Ortega-Arenas, Laura D; Bautista-Martínez, Néstor; Pineda, Samuel

    2015-10-01

    Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) is one of the most economically important pests of potato, tomato, and peppers in Central America, Mexico, the United States, and New Zealand. Its control is based on the use of insecticides; however, recently, the potential of the eulophid parasitoid Tamarixia triozae (Burks) (Hymenoptera: Eulophidae) for population regulation has been studied. Because T. triozae is likely to be exposed to insecticides on crops, the objective of this study was to explore the compatibility of eight insecticides with this parasitoid. The toxicity and residual activity (persistence) of spirotetramat, spiromesifen, beta-cyfluthrin, pymetrozine, azadirachtin, imidacloprid, abamectin, and spinosad against T. triozae adults were assessed using a method based on the residual contact activity of each insecticide on tomato leaf discs collected from treated plants growing under greenhouse conditions. All eight insecticides were toxic to T. triozae. Following the classification of the International Organization of Biological Control, the most toxic were abamectin and spinosad, which could be placed in toxicity categories 3 and 4, respectively. The least toxic were azadirachtin, pymetrozine, spirotetramat, spiromesifen, imidacloprid, and beta-cyfluthrin, which could be placed in toxicity category 2. In terms of persistence, by day 5, 6, 9, 11, 13, 24, and 41 after application, spirotetramat, azadirachtin, spiromesifen, pymetrozine, imidacloprid, beta-cyfluthrin, abamectin, and spinosad could be considered harmless, that is, placed in toxicity category 1 (insecticides allow them to be considered within integrated pest management programs that include T. triozae. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  11. Comparison of the activation energy barrier for succinimide formation from α- and β-aspartic acid residues obtained from density functional theory calculations.

    Science.gov (United States)

    Nakayoshi, Tomoki; Kato, Koichi; Fukuyoshi, Shuichi; Takahashi, Ohgi; Kurimoto, Eiji; Oda, Akifumi

    2018-01-03

    The l-α-Asp residues in peptides or proteins are prone to undergo nonenzymatic reactions to form l-β-Asp, d-α-Asp, and d-β-Asp residues via a succinimide five-membered ring intermediate. From these three types of isomerized aspartic acid residues, particularly d-β-Asp has been widely detected in aging tissue. In this study, we computationally investigated the cyclization of α- and β-Asp residues to form succinimide with dihydrogen phosphate ion as a catalyst (H 2 PO 4 - ). We performed the study using B3LYP/6-31+G(d,p) density functional theory calculations. The comparison of the activation barriers of both residues is discussed. All the calculations were performed using model compounds in which an α/β-Asp-Gly sequence is capped with acetyl and methylamino groups on the N- and C-termini, respectively. Moreover, H 2 PO 4 - catalyzes all the steps of the succinimide formation (cyclization-dehydration) acting as a proton-relay mediator. The calculated activation energy barriers for succinimide formation of α- and β-Asp residues are 26.9 and 26.0kcalmol -1 , respectively. Although it was experimentally confirmed that β-Asp has higher stability than α-Asp, there was no clear difference between the activation barriers. Therefore, the higher stability of β-Asp residue than α-Asp residue may be caused by an entropic effect associated with the succinimide formation. Copyright © 2018 Elsevier B.V. All rights reserved.

  12. MAGIK: a Monte Carlo system for computing induced residual activation dose rates

    International Nuclear Information System (INIS)

    Barish, J.; Gabriel, T.A.; Alsmiller, R.G. Jr.

    1979-08-01

    The photon dose rate from the induced activity produced by sustained bombardment of materials by neutrons and charged particles may present a significant radiation hazard. To minimize this hazard, the material configuration must be so designed that the photon dose rate decays to an acceptable level soon after the source beam is turned off. MAGIK calculates the time-independent photon dose rates that result from activities produced by nucleon-nucleus and meson-nucleus collisions over a wide range of energies. The system has been used both for high-energy accelerator studies and for fusion reactor studies. In the MAGIK system the lengthy photon transport calculations are carried out independent of time, and the time dependence is introduced in the final program, thereby permitting study of various operating scenarios with a minimum computing cost

  13. Pristimerin Inhibits LPS-Triggered Neurotoxicity in BV-2 Microglia Cells Through Modulating IRAK1/TRAF6/TAK1-Mediated NF-κB and AP-1 Signaling Pathways In Vitro.

    Science.gov (United States)

    Hui, Bin; Zhang, Liping; Zhou, Qinhua; Hui, Ling

    2018-02-01

    Microglia plays a prominent role in the brain's inflammatory response to injury or infection by migrating to affected locations and secreting inflammatory molecules. However, hyperactivated microglial is neurotoxic and plays critical roles in the pathogenesis of neurodegenerative diseases. Pristimerin, a naturally occurring triterpenoid, possesses antitumor, antioxidant, and anti-inflammatory activities. However, the effect and the molecular mechanism of pristimerin against lipopolysaccharide (LPS)-induced neurotoxicity in microglia remain to be revealed. In the present study, using BV-2 microglial cultures, we investigated whether pristimerin modifies neurotoxicity after LPS stimulation and which intracellular pathways are involved in the effect of pristimerin. Here we show that pristimerin markedly suppressed the release of Regulated on Activation, Normal T Expressed and Secreted (RANTES), transforming growth factor-β1 (TGF-β1), IL-6, tumor necrosis factor-α (TNF-α), and nitric oxide (NO). Pristimerin also significantly inhibited migration of BV-2 microglia and alleviated the death of neuron-like PC12 cell induced by the conditioned medium from LPS-activated BV-2 microglial cells. Moreover, pristimerin reduced the expression and interaction of TNF Receptor-Associated Factor 6 (TRAF6) and Interleukin-1 Receptor-Associated Kinases (IRAK1), limiting TGF-beta activating kinase 1 (TAK1) activation, and resulting in an inhibition of IKKα/β/NF-κB and MKK7/JNK/AP-1 signaling pathway in LPS-activated BV-2 microglia. Taken together, the anti-neurotoxicity action of pristimerin is mediated through the inhibition of TRAF6/IRAK1/TAK1 interaction as well as the related pathways: IKKα/β/NF-κB and MKK7/JNK/AP-1 signaling pathways. These findings may suggest that pristimerin might serve as a new therapeutic agent for treating hyperactivated microglial induced neurodegenerative diseases.

  14. Trimethyloxonium modification of batrachotoxin-activated Na channels alters functionally important protein residues.

    OpenAIRE

    Cherbavaz, D B

    1995-01-01

    The extracellular side of single batrachotoxin-activated voltage-dependent Na channels isolated from rat skeletal muscle membranes incorporated into neutral planar lipid bilayers were treated in situ with the carboxyl methylating reagent, trimethyloxonium (TMO). These experiments were designed to determine whether TMO alters Na channel function by a general through-space electrostatic mechanism or by methylating specific carboxyl groups essential to channel function. TMO modification reduced ...

  15. DAF in diabetic patients is subject to glycation/inactivation at its active site residues.

    Science.gov (United States)

    Flückiger, Rudolf; Cocuzzi, Enzo; Nagaraj, Ram H; Shoham, Menachem; Kern, Timothy S; Medof, M Edward

    2018-01-01

    Decay accelerating factor (DAF or CD55) is a cell associated C3 and C5 convertase regulator originally described in terms of protection of self-cells from systemic complement but now known to modulate adaptive T cell responses. It is expressed on all cell types. We investigated whether nonenzymatic glycation could impair its function and potentially be relevant to complications of diabetes mellitus and other conditions that result in nonenzymatic glycation including cancer, Alzheimer's disease, and aging. Immunoblots of affinity-purified DAF from erythrocytes of patients with diabetes showed pentosidine, glyoxal-AGEs, carboxymethyllysine, and argpyrimidine. HPLC/MS analyses of glucose modified DAF localized the sites of AGE modifications to K 125 adjacent to K 126 , K 127 at the junction of CCPs2-3 and spatially near R 96 , and R 100 , all identified as being critical for DAF's function. Functional analyses of glucose or ribose treated DAF protein showed profound loss of its regulatory activity. The data argue that de-regulated activation of systemic complement and de-regulated activation of T cells and leukocytes could result from non-enzymatic glycation of DAF. Copyright © 2017. Published by Elsevier Ltd.

  16. Statistical modeling to management and treatment of scrap with low and very low residual activity; Hacia la modelizacion estadistica de la gestion y tratamiento de chatarras con baja y muy bajo actividad residual

    Energy Technology Data Exchange (ETDEWEB)

    Garcia-Bermejo Fernandez, R.; Anaya Lazaro, M.

    2011-07-01

    The experience of recent years on the management of scrap metal containing residual activity have allowed the development of a simple statistical model for the management of these materials. This statistical model includes a breakdown of the various processing operations to which these materials undergo and the effects in the process of radiological controls associated to the control of declassification that defines disposal (recycled by smelting, reclamation, temporary storage the plant or sent to final storage of radioactive waste).

  17. Shelf-life evaluation of Kaṃsaharītakī avaleha and its granules: A preliminary study.

    Science.gov (United States)

    Khemuka, Nidhi; Galib, R; Patgiri, B J; Prajapati, Pradeep Kumar

    2015-01-01

    Savīryatā-avadhi (shelf life) of different Ayurvedic dosage forms is described in Śārṅgadharasaṃhitā. Though the concepts have a strong background, we seek to re-evaluate the age old concepts by following current norms. An attempt has been made in the present study to evaluate shelf-life of Kaṃsaharītakī avaleha and its granules. Raw materials were procured from Pharmacy, Gujarat Ayurved University, Jamnagar and utilized after proper authentication in pharmacognosy lab. The avaleha and its granules were prepared in the departmental laboratory following classical guidelines and subjected to accelerated stability studies. Both the products were found to be free from microbial contamination. Heavy metals were within the prescribed limit. Changes in physico-chemical profiles at different intervals are insignificant. On extrapolation of the observations, the shelf life of avaleha was found to be18 months and 27 months to the granules. Stability of granules was found to be comparatively higher than the avaleha. This observation may be exclusive to Kaṃsaharītakī avaleha. Studies on other avalehas and their granules need to be carried out to confirm this preliminary observation.

  18. Degradation of flumequine by the Fenton and photo-Fenton processes: Evaluation of residual antimicrobial activity

    International Nuclear Information System (INIS)

    Rodrigues-Silva, Caio; Maniero, Milena Guedes; Rath, Susanne; Guimarães, José Roberto

    2013-01-01

    Flumequine is a broad-spectrum antimicrobial agent of the quinolone class, and it is widely used as a veterinary drug in food-producing animals. The presence of flumequine in the environment may contribute to the development of drug resistant bacterial strains. In this study, water samples fortified with flumequine (500 μg L −1 ) were degraded using the Fenton and photo-Fenton processes. The maximum degradation efficiency for flumequine by the Fenton process was approximately 40% (0.5 mmol L −1 Fe(II), 2.0 mmol L −1 H 2 O 2 and 15 min). By applying UV radiation (photo-Fenton process), the efficiency reached more than 94% in 60 min when 0.25 mmol L −1 Fe(II) and 10.0 mmol L −1 H 2 O 2 were used. Under these conditions, the Fenton process was able to reduce the biological activity, whereas the photo-Fenton process eliminated almost all of the antimicrobial activity because it was not detected. Four byproducts with an m/z of 244, 238, 220 and 202 were identified by mass spectrometry, and a degradation pathway for flumequine was proposed. The byproducts were derived from decarboxylation and defluorination reactions and from modifications in the alkylamino chain of the fluoroquinolone. - Highlights: ► Photo-Fenton process achieved the maximum performance, degrading 94% of flumequine. ► As the flumequine concentration decreased, antimicrobial activity also decreased. ► Four byproducts with m/z of 244, 238, 220 and 202 were identified. ► A degradation pathway for flumequine was proposed

  19. Degradation of flumequine by the Fenton and photo-Fenton processes: Evaluation of residual antimicrobial activity

    Energy Technology Data Exchange (ETDEWEB)

    Rodrigues-Silva, Caio; Maniero, Milena Guedes [School of Civil Engineering, Architecture and Urbanism, University of Campinas — UNICAMP, P.O. Box 6021, CEP 13083-852, Campinas, SP (Brazil); Rath, Susanne [Chemistry Institute, University of Campinas — UNICAMP, P.O. Box 6154, CEP 13084-971, Campinas, SP (Brazil); Guimarães, José Roberto, E-mail: jorober@fec.unicamp.br [School of Civil Engineering, Architecture and Urbanism, University of Campinas — UNICAMP, P.O. Box 6021, CEP 13083-852, Campinas, SP (Brazil)

    2013-02-15

    Flumequine is a broad-spectrum antimicrobial agent of the quinolone class, and it is widely used as a veterinary drug in food-producing animals. The presence of flumequine in the environment may contribute to the development of drug resistant bacterial strains. In this study, water samples fortified with flumequine (500 μg L{sup −1}) were degraded using the Fenton and photo-Fenton processes. The maximum degradation efficiency for flumequine by the Fenton process was approximately 40% (0.5 mmol L{sup −1} Fe(II), 2.0 mmol L{sup −1} H{sub 2}O{sub 2} and 15 min). By applying UV radiation (photo-Fenton process), the efficiency reached more than 94% in 60 min when 0.25 mmol L{sup −1} Fe(II) and 10.0 mmol L{sup −1} H{sub 2}O{sub 2} were used. Under these conditions, the Fenton process was able to reduce the biological activity, whereas the photo-Fenton process eliminated almost all of the antimicrobial activity because it was not detected. Four byproducts with an m/z of 244, 238, 220 and 202 were identified by mass spectrometry, and a degradation pathway for flumequine was proposed. The byproducts were derived from decarboxylation and defluorination reactions and from modifications in the alkylamino chain of the fluoroquinolone. - Highlights: ► Photo-Fenton process achieved the maximum performance, degrading 94% of flumequine. ► As the flumequine concentration decreased, antimicrobial activity also decreased. ► Four byproducts with m/z of 244, 238, 220 and 202 were identified. ► A degradation pathway for flumequine was proposed.

  20. Preliminary results of MR imaging of lymphoma: Distinguishing active tumor from benign residue

    International Nuclear Information System (INIS)

    Drace, J.; Baker, L.L.; Chang, P.; Castellino, R.A.

    1987-01-01

    Distinguishing tumor from benign posttreatment tissue based on both morphologic and tissue characteristics is critically important. Patients are studied before, during, and after treatment; at the time of recurrence; and on long-term follow-up. Multisection spin-echo sequences in orthogonal planes and a special single-section tissue characterization matrix of 16 different repetition time/echo time combinations are used. These basic images are used for cluster analysis (approximate fuzzy C means), T1-T2 synthetic images, linear combinations, and comparison with internal standards. Preliminary results in 35 patients imaged before treatment and 12 patients with follow-up examinations consistently show lymphoma masses to have complex architecture with high T2-weighted signal and moderate T1-weighted signal, distinct from posttreatment fibrosis. Uncommon components of active tumor with low T2-weighted signal appear distinct from fibrosis on T1-weighted images. Preliminary cluster analysis results show distinct clustering of active lymphoma versus fibrosis and biopsy-proved cystic degeneration

  1. Residuation theory

    CERN Document Server

    Blyth, T S; Sneddon, I N; Stark, M

    1972-01-01

    Residuation Theory aims to contribute to literature in the field of ordered algebraic structures, especially on the subject of residual mappings. The book is divided into three chapters. Chapter 1 focuses on ordered sets; directed sets; semilattices; lattices; and complete lattices. Chapter 2 tackles Baer rings; Baer semigroups; Foulis semigroups; residual mappings; the notion of involution; and Boolean algebras. Chapter 3 covers residuated groupoids and semigroups; group homomorphic and isotone homomorphic Boolean images of ordered semigroups; Dubreil-Jacotin and Brouwer semigroups; and loli

  2. Elimination of microcystin-LR and residual Mn species using permanganate and powdered activated carbon: Oxidation products and pathways.

    Science.gov (United States)

    Jeong, Boyoung; Oh, Min-Seok; Park, Hyun-Mee; Park, Chanhyuk; Kim, Eun-Ju; Hong, Seok Won

    2017-05-01

    The oxidation of microcystin-LR (MC-LR) in deionized water (DI) and river water using potassium permanganate (KMnO 4 ) at a neutral pH and at 23 ± 2 °C was investigated. These two aqueous systems (i.e., DI and river water) gave comparable second-order rate constants (289.9 and 285.5 M -1 s -1 (r 2  > 0.99), respectively), which confirmed the effectiveness of this oxidation process for the treatment of natural surface water. The presence of either humic or fulvic acid reduced the removal efficiency of MC-LR, with the latter exhibiting a greater inhibitory effect. Monitoring of MC-LR and residual Mn 2+ levels with adding KMnO 4 (1 mg/L) and powdered activated carbon (PAC, 5-20 mg L -1 ) before and during coagulation, respectively, revealed that 60 min of permanganate pre-oxidation followed by coagulant addition with PAC was the most effective approach for reducing both levels below limits stated by WHO guidelines. The MC-LR degradation products were the result of oxidation occurring at the diene and aromatic moieties of the Adda (3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid) side-chain, in addition to amine bond hydrolysis of the Mdha (N-methyldehydroalanine) moiety. Several toxic by-products with an intact Adda chain were observed during the reaction, but completely disappeared after 60 min. This further supports the conclusion that sufficient contact time with permanganate (i.e., >60 min) is essential to reducing the residual toxicity and maximizing the efficiency of MC-LR oxidation when treating raw water. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Removal of residual functionalized ionic liquids from water by ultrasound-assisted zero-valent iron/activated carbon.

    Science.gov (United States)

    Zhou, Haimei; Lv, Ping; Qi, Hang; Ma, Jinqi; Wang, Jianji

    2018-03-02

    Numerous applications of ionic liquids (ILs) are often accompanied by the generation of aqueous wastes. Due to the high toxicity and poor biodegradability of ILs, effective chemical treatment is of great importance for their removal from aqueous solution. In this work, an ultrasound-assisted zero-valent iron/activated carbon (US-ZVI/AC) micro-electrolysis technique was used to degrade residual functionalized ILs, 1-butyl-3-methyl benzimidazolium bromide ([BMBIM]Br) and 1-allyl-3-methylimidazolium chloride ([AMIM]Cl) in aqueous solution, and the degradation degree, degradation kinetics and possible degradation pathways were investigated. It was shown that the degradation of these functionalized ILs was highly efficient in the US-ZVI/AC system, and the degradation degree was as high as 96.1% and 92.9% in 110 min for [BMBIM]Br and [AMIM]Cl, respectively. The degradation of [BMBIM]Br could be described by the second-order kinetics model, and [BMBIM] + was decomposed in two ways: (i) sequential cleavage of N-alkyl side chain of the cation produced three intermediates; (ii) the 2-positioned H atoms of the benzimidazolium ring were first oxidized, and then the imidazolium ring was opened. The degradation of [AMIM]Cl followed the first-order kinetics rule, and the 2,4,5-positioned H atoms of the imidazolium ring were oxidized to induce ring opening. In addition, the removal of total organic carbon was found to be >87%, which indicates that most of the ILs was mineralized in the degradation process. These results suggest that ultrasound-assisted ZVI/AC micro-electrolysis is highly effective for the removal of residual functionalized ILs from aqueous environment.

  4. Regulated internalization of NMDA receptors drives PKD1-mediated suppression of the activity of residual cell-surface NMDA receptors.

    Science.gov (United States)

    Fang, Xiao-Qian; Qiao, Haifa; Groveman, Bradley R; Feng, Shuang; Pflueger, Melissa; Xin, Wen-Kuan; Ali, Mohammad K; Lin, Shuang-Xiu; Xu, Jindong; Duclot, Florian; Kabbaj, Mohamed; Wang, Wei; Ding, Xin-Sheng; Santiago-Sim, Teresa; Jiang, Xing-Hong; Salter, Michael W; Yu, Xian-Min

    2015-11-19

    Constitutive and regulated internalization of cell surface proteins has been extensively investigated. The regulated internalization has been characterized as a principal mechanism for removing cell-surface receptors from the plasma membrane, and signaling to downstream targets of receptors. However, so far it is still not known whether the functional properties of remaining (non-internalized) receptor/channels may be regulated by internalization of the same class of receptor/channels. The N-methyl-D-aspartate receptor (NMDAR) is a principal subtype of glutamate-gated ion channel and plays key roles in neuronal plasticity and memory functions. NMDARs are well-known to undergo two types of regulated internalization - homologous and heterologous, which can be induced by high NMDA/glycine and DHPG, respectively. In the present work, we investigated effects of regulated NMDAR internalization on the activity of residual cell-surface NMDARs and neuronal functions. In electrophysiological experiments we discovered that the regulated internalization of NMDARs not only reduced the number of cell surface NMDARs but also caused an inhibition of the activity of remaining (non-internalized) surface NMDARs. In biochemical experiments we identified that this functional inhibition of remaining surface NMDARs was mediated by increased serine phosphorylation of surface NMDARs, resulting from the activation of protein kinase D1 (PKD1). Knockdown of PKD1 did not affect NMDAR internalization but prevented the phosphorylation and inhibition of remaining surface NMDARs and NMDAR-mediated synaptic functions. These data demonstrate a novel concept that regulated internalization of cell surface NMDARs not only reduces the number of NMDARs on the cell surface but also causes an inhibition of the activity of remaining surface NMDARs through intracellular signaling pathway(s). Furthermore, modulating the activity of remaining surface receptors may be an effective approach for treating receptor

  5. Optimization of analitycal control over residues of active ingridients of modern pesticides in reservoirs water

    Directory of Open Access Journals (Sweden)

    Semenenko V.M.

    2013-10-01

    Full Text Available A highly sensitive and selective method of pyraclostrobin, boscalid, tebufenpyrad and prohexadione-calcium determination under their combined presence in water sample, using high-performance liquid chromatography was developed. On the base of mentioned active ingredients (combined fungicide Bellis, insecto-acaricide Masai and plant growth regulator Regalis pesticides may be used in one vegetation season for fruit trees protection. Method of co-determination of these substances is based on the preparation of water samples for extraction, extraction of pyraclostrobin, boscalid, tebufenpyrad and prohexadione-calcium, concentrating of extract of substances mixtures and chromatographic determination with ultraviolet detection. A distinctive feature of this method is changing of ratio of components of mobile phase (mixture of acetonitrile and 0,1 % aqueous solution of phosphoric acid in the process of chromatographic analysis, which allowed to clearly visualize test substances in case of their joint presence in one sample. Implementation of developed and patented method into practice optimizes control over application of pesticides in agriculture and their monitoring in reservoirs water by significant acceleration of analysis and reduction of expenses in its carying out.

  6. Effect of repeated pesticide applications on soil properties in cotton fields: II. Insecticide residues and impact on dehydrogenase and arginine deaminase activities

    International Nuclear Information System (INIS)

    Vig, K.; Singh, D.K.; Agarwal, H.C.; Dhawan, A.K.; Dureja, P.

    2001-01-01

    Insecticides were applied sequentially at recommended dosages post crop emergence in cotton fields and soil was sampled at regular intervals after each treatment. Soil was analysed for insecticide residues and activity of the enzymes dehydrogenase and arginine deaminase. Insecticide residues detected in the soil were in small quantities and they did not persist for long. Only endosulfan leached below 15 cm. Insecticides had only temporary effects on enzyme activities which disappeared either before the next insecticide treatment or by the end of the experimental period. (author)

  7. Larvicidal activity of Myrtaceae essential oils and their components against Aedes aegypti, acute toxicity on Daphnia magna, and aqueous residue.

    Science.gov (United States)

    Park, Hye-Mi; Kim, Junheon; Chang, Kyu-Sik; Kim, Byung-Seok; Yang, Yu-Jung; Kim, Gil-Hah; Shin, Sang-Chul; Park, Il-Kwon

    2011-03-01

    The larvicidal activity of 11 Myrtaceae essential oils and their constituents was evaluated against Aedes aegypti L. Of the 11, Melaleuca linariifolia Sm., Melaleuca dissitiflora F. Muell., Melaleuca quinquenervia (Cav.) S. T. Blake, and Eucalyptus globulus Labill oils at 0.1 mg/ml exhibited > or = 80% larval mortality. At this same concentration, the individual constituents tested, allyl isothiocyanate, alpha-terpinene, p-cymene, (+)-limonene, (-)-limonene, gamma-terpinene, and (E)-nerolidol, resulted in > or = 95% mortality. We also tested the acute toxicity of these four active oils earlier mentioned and their constituents against Daphnia magna Straus. M. linariifolia and allyl isothiocyanate was the most toxic to D. magna. Twodays after treatment, residues of M. dissitiflora, M. linariifolia, M. quinquenervia, and E. globulus oils in water were 55.4, 46.6, 32.4, and 14.8%, respectively. Less than 10% of allyl isothiocyanate, alpha-terpinene, p-cymene, (-)-limonene, (+)-limonene, and gamma-terpinene was detected in the water at 2 d after treatment. Our results indicated that oils and their constituents could easily volatilize in water within a few days after application, thus minimizing their effect on the aqueous ecosystem. Therefore, Myrtaceae essential oils and their constituents could be developed as control agents against mosquito larvae.

  8. Residual β activity of particulate 234Th as a novel proxy for tracking sediment resuspension in the ocean

    Science.gov (United States)

    Lin, Wuhui; Chen, Liqi; Zeng, Shi; Li, Tao; Wang, Yinghui; Yu, Kefu

    2016-06-01

    Sediment resuspension occurs in the global ocean, which greatly affects material exchange between the sediment and the overlying seawater. The behaviours of carbon, nutrients, heavy metals, and other pollutants at the sediment-seawater boundary will further link to climate change, eutrophication, and marine pollution. Residual β activity of particulate 234Th (RAP234) is used as a novel proxy to track sediment resuspension in different marine environments, including the western Arctic Ocean, the South China Sea, and the Southern Ocean. Sediment resuspension identified by high activity of RAP234 is supported by different lines of evidence including seawater turbidity, residence time of total 234Th, Goldschmidt’s classification, and ratio of RAP234 to particulate organic carbon. A conceptual model is proposed to elucidate the mechanism for RAP234 with dominant contributions from 234Th-238U and 212Bi-228Th. The ‘slope assumption’ for RAP234 indicated increasing intensity of sediment resuspension from spring to autumn under the influence of the East Asian monsoon system. RAP234 can shed new light on 234Th-based particle dynamics and should benefit the interpretation of historical 234Th-238U database. RAP234 resembles lithophile elements and has broad implications for investigating particle dynamics in the estuary-shelf-slope-ocean continuum and linkage of the atmosphere-ocean-sediment system.

  9. The essential role of the sialic acid residues for IFN-β1a activity determined in vivo

    DEFF Research Database (Denmark)

    Olesen, Lasse Dissing; Andersen, Morten Thaysen

    the function of the rhIFN-β1a glycan moiety and its individual carbohydrate residues using the myxovirus resistance (Mx) mRNA as a biomarker in Mx-congenic mice. We showed that the Mx mRNA level in blood leukocytes peaked three hours after subcutaneous administration of rhIFN-β1a. In addition, a clear dose......-response relationship was confirmed and the Mx response was shown to be receptor mediated. Using specific glycosidases, different glycosylation analogs of rhIFN-β1a were obtained and their activities determined. The glycosylated rhIFN-β1a showed significantly higher activity than its deglycosylated counterpart, due...... to a protein stabilization/solubilization effect of the glycan. Interestingly, the terminating sialic acids were essential for these effects. Conclusively, the structure/bioactivity relationship of rhIFN-β1a was determined in vivo and provided a novel insight into the role of the rhIFN-β1a glycan and its...

  10. Indoor residual spraying with a mixture of clothianidin (a neonicotinoid insecticide) and deltamethrin provides improved control and long residual activity against pyrethroid resistant Anopheles gambiae sl in Southern Benin.

    Science.gov (United States)

    Ngufor, Corine; Fongnikin, Augustin; Rowland, Mark; N'Guessan, Raphael

    2017-01-01

    There is an urgent need for new insecticides for indoor residual spraying (IRS) which can provide improved and prolonged control of malaria vectors that have developed resistance to existing insecticides. The neonicotinoid, clothianidin represents a class of chemistry new to public health. Clothianidin acts as an agonist on nicotinic acetyl choline receptors. IRS with a mixture of Clothianidin and another WHO approved insecticide such as deltamethrin could provide improved control of insecticide resistant malaria vector populations and serve as a tool for insecticide resistance management. The efficacy and residual activity of a novel IRS mixture of deltamethrin and clothianidin was evaluated against wild pyrethroid resistant An. gambiae sl in experimental huts in Cove, Benin. Two application rates of the mixture were tested and comparison was made with clothianidin and deltamethrin applied alone. To assess the residual efficacy of the treatments on different local wall substrates, the inner walls of the experimental huts were covered with either cement, mud or plywood. Clothianidin demonstrated a clear delayed expression in mortality of wild pyrethroid resistant An. gambiae sl in the experimental huts which reached its full effect 120 hours after exposure. Overall mortality over the 12-month hut trial was 15% in the control hut and 24-29% in the deltamethrin-treated huts. The mixture of clothianidin 200mg/m2 and deltamethrin 25mg/m2 induced high overall hut mortality rates (87% on mud walls, 82% on cement walls and 61% on wooden walls) largely due to the clothianidin component and high hut exiting rates (67-76%) mostly due to the deltamethrin component. Mortality rates remained >80% for 8-9 months on mud and cement walls. The residual activity trend was confirmed by results from monthly in situ cone bioassays with laboratory susceptible An. gambiae Kisumu strain. IRS campaigns with the mixture of clothianidin plus deltamethrin have the potential to provide

  11. Indoor residual spraying with a mixture of clothianidin (a neonicotinoid insecticide and deltamethrin provides improved control and long residual activity against pyrethroid resistant Anopheles gambiae sl in Southern Benin.

    Directory of Open Access Journals (Sweden)

    Corine Ngufor

    Full Text Available There is an urgent need for new insecticides for indoor residual spraying (IRS which can provide improved and prolonged control of malaria vectors that have developed resistance to existing insecticides. The neonicotinoid, clothianidin represents a class of chemistry new to public health. Clothianidin acts as an agonist on nicotinic acetyl choline receptors. IRS with a mixture of Clothianidin and another WHO approved insecticide such as deltamethrin could provide improved control of insecticide resistant malaria vector populations and serve as a tool for insecticide resistance management.The efficacy and residual activity of a novel IRS mixture of deltamethrin and clothianidin was evaluated against wild pyrethroid resistant An. gambiae sl in experimental huts in Cove, Benin. Two application rates of the mixture were tested and comparison was made with clothianidin and deltamethrin applied alone. To assess the residual efficacy of the treatments on different local wall substrates, the inner walls of the experimental huts were covered with either cement, mud or plywood.Clothianidin demonstrated a clear delayed expression in mortality of wild pyrethroid resistant An. gambiae sl in the experimental huts which reached its full effect 120 hours after exposure. Overall mortality over the 12-month hut trial was 15% in the control hut and 24-29% in the deltamethrin-treated huts. The mixture of clothianidin 200mg/m2 and deltamethrin 25mg/m2 induced high overall hut mortality rates (87% on mud walls, 82% on cement walls and 61% on wooden walls largely due to the clothianidin component and high hut exiting rates (67-76% mostly due to the deltamethrin component. Mortality rates remained >80% for 8-9 months on mud and cement walls. The residual activity trend was confirmed by results from monthly in situ cone bioassays with laboratory susceptible An. gambiae Kisumu strain.IRS campaigns with the mixture of clothianidin plus deltamethrin have the potential to

  12. Chemometric comparison of polychlorinated biphenyl residues and toxicologically active polychlorinated biphenyl congeners in the eggs of Forster's Terns (Sterna fosteri)

    Science.gov (United States)

    Schwartz, Ted R.; Stalling, David L.

    1991-01-01

    The separation and characterization of complex mixtures of polychlorinated biphenyls (PCBs) is approached from the perspective of a problem in chemometrics. A technique for quantitative determination of PCB congeners is described as well as an enrichment technique designed to isolate only those congener residues which induce mixed aryl hydrocarbon hydroxylase enzyme activity. A congener-specific procedure is utilized for the determination of PCBs in whichn-alkyl trichloroacetates are used as retention index marker compounds. Retention indices are reproducible in the range of ±0.05 to ±0.7 depending on the specific congener. A laboratory data base system developed to aid in the editing and quantitation of data generated from capillary gas chromatography was employed to quantitate chromatographic data. Data base management was provided by computer programs written in VAX-DSM (Digital Standard MUMPS) for the VAX-DEC (Digital Equipment Corp.) family of computers.In the chemometric evaluation of these complex chromatographic profiles, data are viewed from a single analysis as a point in multi-dimensional space. Principal Components Analysis was used to obtain a representation of the data in a lower dimensional space. Two-and three-dimensional proections based on sample scores from the principal components models were used to visualize the behavior of Aroclor® mixtures. These models can be used to determine if new sample profiles may be represented by Aroclor profiles. Concentrations of individual congeners of a given chlorine substitution may be summed to form homologue concentration. However, the use of homologue concentrations in classification studies with environmental samples can lead to erroneous conclusions about sample similarity. Chemometric applications are discussed for evaluation of Aroclor mixture analysis and compositional description of environmental residues of PCBs in eggs of Forster's terns (Sterna fosteri) collected from colonies near Lake Poygan

  13. Atividade residual de herbicidas aplicados ao solo em relação ao controle de quatro espécies de Amaranthus Residual activity of herbicides applied to the soil in relation to control of four Amaranthus Species

    Directory of Open Access Journals (Sweden)

    M.A. Raimondi

    2010-01-01

    -metolachlor foram eficientes para todas as espécies até 30 DAA, em ambas as doses, demonstrando atividade residual consistente para o solo estudado.Herbicides applied at pre-emergence normally present residual activity in the soil, controlling the first weed germinations, and preventing initial weed competition. The aim of this study was to determine the period of residual activity provided by sufficient herbicide rates for 95% (C95 control of the species Amaranthus hybridus, A. lividus, A. spinosus and A. viridis, and to assess the recommended rates of these herbicides. The study was conducted under greenhouse conditions in soils of sandy clay loam texture (20% clay and 1.9 of organic matter, with rates of alachlor, diuron, oxyfluorfen, pendimethalin, prometryne, oxyfluorfen, S-metolachlor, trifluralin 450 and trifluralin 600 being applied at 30, 20, 10 and 0 days before weed sowing. Weed control was evaluated after herbicide permanence in soil for 0, 10, 20 and 30 days after application (DAA. The residual activity of alachlor and prometryne at a rate C95 was not sufficient for the efficient control (>80% of the species for up to 30 DAA. For alachlor, the use of the recommended rate did not provide considerable increase in residual activity, except for A. viridis. The recommended rate of prometryn promoted efficient control of the species for up to 30 DAA, except for A. hybridus. The recommended rate of oxyfluorfen provided an efficient control of A. hybridus and A. spinosus up to 30 DAA. These species had not been effectively controlled by the rate C95. Trifluralin 450 promoted efficient residual control for up to 30 DAA only compared with A. hybridus. Trifluralin 600 was effective in controlling A. hybridus and A. viridis up to 30 DAA and 29 and 28 DAA for A. lividus and A. spinosus, respectively. Clomazone did not promote efficient control of the species for 30 DAA, except for A. viridis. Diuron, pendimethalin and S-metolachlor were effective in controlling the species for up

  14. Identification of tyrosine residues in the intracellular domain of the growth hormone receptor required for transcriptional signaling and Stat5 activation

    DEFF Research Database (Denmark)

    Hansen, L. H.; Wang, X.; Kopchick, J J

    1996-01-01

    phosphorylation in intracellular signaling, we constructed GH receptors in which combinations of tyrosines were mutated to phenylalanines. We identified three tyrosine residues at positions 534, 566, and 627 that were required for activation of GH-stimulated transcription of the serine protease inhibitor (Spi) 2...

  15. Correlation of loss of activity of human aldehyde dehydrogenase with reaction of bromoacetophenone with glutamic acid-268 and cysteine-302 residues. Partial-sites reactivity of aldehyde dehydrogenase.

    Science.gov (United States)

    Abriola, D P; MacKerell, A D; Pietruszko, R

    1990-01-01

    Bromoacetophenone (2-bromo-1-phenylethanone) has been characterized as an affinity reagent for human aldehyde dehydrogenase (EC 1.2.1.3) [MacKerell, MacWright & Pietruszko (1986) Biochemistry 25, 5182-5189], and has been shown to react specifically with the Glu-268 residue [Abriola, Fields, Stein, MacKerell & Pietruszko (1987) Biochemistry 26, 5679-5684] with an apparent inactivation stoichiometry of two molecules of bromoacetophenone per molecule of enzyme. The specificity of bromoacetophenone for reaction with Glu-268, however, is not absolute, owing to the extreme reactivity of this reagent. When bromo[14C]acetophenone was used to label the human cytoplasmic E1 isoenzyme radioactively and tryptic fragmentation was carried out, peptides besides that containing Glu-268 were found to have reacted with reagent. These peptides were purified by h.p.l.c. and analysed by sequencing and scintillation counting to quantify radioactive label in the material from each cycle of sequencing. Reaction of bromoacetophenone with the aldehyde dehydrogenase molecule during enzyme activity loss occurs with two residues, Glu-268 and Cys-302. The activity loss, however, appears to be proportional to incorporation of label at Glu-268. The large part of incorporation of label at Cys-302 occurs after the activity loss is essentially complete. With both Glu-268 and Cys-302, however, the incorporation of label stops after one molecule of bromoacetophenone has reacted with each residue. Reaction with other residues continues after activity loss is complete. PMID:1968743

  16. The Role of Active Site Residues in ATP Binding and Catalysis in the Methanosarcina thermophila Acetate Kinase

    Directory of Open Access Journals (Sweden)

    Cheryl Ingram-Smith

    2015-03-01

    Full Text Available Acetate kinase (ACK, which catalyzes the reversible phosphorylation of acetate by ATP, is a member of the acetate and sugar kinase/heat shock cognate/actin (ASKHA superfamily. ASKHA family members share a common core fold that includes an ATPase domain with five structural motifs. The PHOSPHATE1 motif has previously been shown to be important for catalysis. We have investigated the role of two of these motifs in the Methanosarcina thermophila ACK (MtACK and have shown that residues projecting into the ACK active site from the PHOSPHATE2 and ADENOSINE loops and a third highly conserved loop designated here as LOOP3 play key roles in nucleotide triphosphate (NTP selection and utilization. Alteration of Asn211 of PHOSPHATE2, Gly239 of LOOP3, and Gly331 of ADENOSINE greatly reduced catalysis. In particular, Gly331, which is highly conserved throughout the ASKHA superfamily, has the greatest effect on substrate selection. Alteration at this site strongly skewed MtACK toward utilization of purines over pyrimidines, unlike the wild type enzyme that shows broad NTP utilization. Further investigation into differences between the ATPase domain in MtACK and other acetate kinases that show different substrate preferences will provide us with a better understanding of the diversity of phosphoryl donor selection in this enzyme family.

  17. Effect of different extracting solvents on antioxidant activity and phenolic compounds of a fruit and vegetable residue flour

    Directory of Open Access Journals (Sweden)

    Mônica C. P. Santos

    2016-01-01

    Full Text Available In order to quantify antioxidant capacity in food products, several methods have been proposed over the years. Among them, DPPH radical is widely used to determine the antioxidant capacity of different substrates. However, it is known that different types of extractants, providing different responses, can extract a variety of bioactive compounds. Besides, storage time seems to interfere in the stability of these substances. Integral use of fruits and vegetables has been proposed along the years as a means of reducing environmental pollution and give a better destination to by-products from food industries. Thus, this study aimed to evaluate the antioxidant potential of a fruit and vegetables residue flour (FVR with sequential and non-sequential extraction, in order to evaluate its antioxidant activity and phenolic compounds. And these compounds stability during storage of 180 days. It was observed that in non-sequential extraction, water was able to reduce by 74% the radical; however, at sequential extraction process, using six different extractors, each one was able to reduce at least 40% of DPPH. The total soluble phenolic contents in sequential extraction were 22.49 ± 1.59 mg GAE/g FVR on the first day and 5.35 ± 0.32 mg GAE/g FVR after 180 days.

  18. Evidence in Latin America of recurrence of V388M, a phenylketonuria mutation with high in vitro residual activity

    Energy Technology Data Exchange (ETDEWEB)

    Desviat, L.R.; Perez, B.; De Lucca, M. [Universidad Autonoma de Madrid, (Spain)] [and others

    1995-08-01

    Phenylketonuria mutation V388M is frequent in the Iberian Peninsula. In vitro, the V388M mutant enzyme has similar immunoreactive protein and phenylalanine hydroxylase mRNA and had 43% residual activity, which correlates well with the mild phenotype exhibited by the homozygous patients. In Spain it has been detected in 5.7% of the mutant alleles and is always associated with haplotype 1.7. This mutation is also present in high frequency in some Latin American countries (Brazil, 9% Chile, 13%). It is interesting that in Chile most of the alleles bearing this mutation carry haplotype 4.3, although in Brazil it is found only on the background of haplotype 1.7. The origin of V388M in Spain on haplotype 1.7 and in Chile on haplotype 4.3 is clearly different. Recurrence is the most plausible explanation, because the mutation involves a CpG dinucleotide, and a recombination event transferring the mutation from haplotype 1 to 4 is unlikely. 29 refs., 2 figs., 3 tabs.

  19. The Catalytic Activity of Modified Zeolite Lanthanum on the Catalytic Cracking of Al-Duara Atmospheric Distillation Residue

    Directory of Open Access Journals (Sweden)

    Karim Khalifa Esgair

    2016-03-01

    Full Text Available Atmospheric residue fluid catalytic cracking was selected as a probe reaction to test the catalytic performance of modified NaY zeolites and prepared NaY zeolites. Modified NaY zeolites have been synthesized by simple ion exchange methods. Three samples of modified zeolite Y have been obtained by replacing the sodium ions in the original sample with lanthanum and the weight percent added are 0.28, 0.53, and 1.02 respectively. The effects of addition of lanthanum to zeolite Y in different weight percent on the cracking catalysts were investigated using an experimental laboratory plant scale of fluidized bed reactor. The experiments have been performed with weight hourly space velocity (WHSV range of 6 to 24 h-1, and the range of temperature from 450 to 510 oC. The activity of the catalyst with 1.02 wt% lanthanum has been shown to be much greater than that of the sample parent NaY. Also it was observed that the addition of the lanthanum causes an increase in the thermal stability of the zeolite.

  20. CXC195 suppresses proliferation and inflammatory response in LPS-induced human hepatocellular carcinoma cells via regulating TLR4-MyD88-TAK1-mediated NF-κB and MAPK pathway

    International Nuclear Information System (INIS)

    Wang, Yiting; Tu, Qunfei; Yan, Wei; Xiao, Dan; Zeng, Zhimin; Ouyang, Yuming; Huang, Long; Cai, Jing; Zeng, Xiaoli; Chen, Ya-Jie; Liu, Anwen

    2015-01-01

    Highlights: • CXC195 exhibited significant anti-proliferative effect and induced cell cycle arrest in LPS-induced HepG2 cells. • CXC195 suppressed the release of pro-inflammatory mediators in LPS-induced HepG2 cells. • CXC195 regulated TLR4-MyD88-TAK1-mediated NF-κB and MAPK pathway in LPS-induced HepG2 cells. - Abstract: CXC195 showed strong protective effects in neuronal apoptosis by exerting its antioxidant activity. However, the anti-cancer effects of CXC195 is still with limited acquaintance. Here, we investigated the role of CXC195 in lipopolysaccharide (LPS)-induced human hepatocellular carcinoma (HCC) cells lines (HepG2) and the possible signaling pathways. CXC195 exhibited significant anti-proliferative effect and induced cell cycle arrest in LPS-induced HepG2 cells. In addition, CXC195 suppressed the release of pro-inflammatory mediators in LPS-induced HepG2 cells, including TNF-α, iNOS, IL-1β, IL-6, CC chemokine ligand (CCL)-2, CCL-22 and epidermal growth factor receptor (EGFR). Moreover, CXC195 inhibited the expressions and interactions of TLR4, MyD88 and TAK1, NF-κB translocation to nucleus and its DNA binding activity, phosphorylation of ERK1/2, p38 and JNK. Our results suggested that treatment with CXC195 could attenuate the TLR4-mediated proliferation and inflammatory response in LPS-induced HepG2 cells, thus might be beneficial for the treatment of HCC

  1. Sastra dan Difabel: Menilik Citra Difabel dalam Novel Biola Tak Berdawai dari Sudut Pandang Sosiologi Sastra Ian Watt

    Directory of Open Access Journals (Sweden)

    Mukhanif Yasin Yusuf

    2015-06-01

    Dengan menggunakan pendekatan teori sosiologi sastra Ian Watt, Biola Tak Berdawai tidak jauh berbeda dengan karya-karya Seno lainnya yang melemparkan gagasan kritis terhadap realitas sosial. Latar belakang sosial Seno yang sekaligus sebagai wartawan melemparkan gagasan kritisnya terkait kondisi difabel yang masih mendapat stigma negatif dari masyarakat. Cerminan sosial dalam novel tidak jauh berbeda dengan realitas yang terjadi di Indonesia, dimana ideologi kenormalan menyumbangkan berbagai bentuk ketidakadilan terhadap difabel. Difabel masih dianggap sebagai individu yang cacat, sebagai kutukan Tuhan, dan sebagai sumber aib bagi keluarga. Pendobrakan terhadap realitas yang ada, dilakukan pengarang lewat tokoh utama “Aku” yang difabel dengan menyajikan fakta bahwa difabel memiliki kemampuan yang berbeda, tetapi masyarakat masih belum memahaminya karena sudah terlanjur terjebak pada stigma negatif terhadap difabel.

  2. Active disease and residual damage in treated Wegener's granulomatosis: an observational study using pulmonary high-resolution computed tomography

    Energy Technology Data Exchange (ETDEWEB)

    Komocsi, Andras [Poliklinik fuer Rheumatologie, Universitaet Luebeck, Ratzeburger Allee 160, 23538 Luebeck (Germany); Reuter, Michael; Heller, Martin [Klinik fuer Diagnostische Radiologische, Universitaet Kiel, Arnold-Heller-Strasse 7, 24105 Kiel (Germany); Murakoezi, Henriette; Gross, Wolfgang L.; Schnabel, Armin [Poliklinik fuer Rheumatologie, Universitaet Luebeck, Ratzeburger Allee 160, 23538 Luebeck (Germany); Rheumaklinik Bad Bramstedt, Ratzeburger Allee 160, 23538 Luebeck (Germany)

    2003-01-01

    The purpose of this study was to determine to what extent high-resolution computed tomography (HRCT) of the lungs can distinguish active inflammatory disease from inactive cicatricial disease in patients treated for Wegener's granulomatosis (WG). Twenty-eight WG patients with active pulmonary disease underwent a first HRCT examination immediately before standard immunosuppressive treatment and a second examination after clinical remission had been achieved. Lesions remaining after treatment were categorized as residual damage and were compared with findings during active disease to see by what features active and cicatricial disease can be distinguished. During active disease 17 patients had nodules/masses, 12 had ground-glass opacities, 6 had septal lines and 6 had non-septal lines. After treatment, ground-glass opacities had resolved completely. Nodules/masses had resolved in 8 patients and had diminished in 7 patients. Residual nodules were distinguished from nodules/masses in active disease by lack of cavitation and a diameter of mostly <15 mm. In one-third of patients lines resolved, but in 8 instances new lines evolved during immunosuppression. During a follow-up period of a median 26.5 months (range 20.0-33.8), patients with residual nodules or lines had no more relapses than patients with completely cleared lungs. Treated pulmonary WG leaves substantial residual damage. High-resolution CT does assist in the distinction between active and inactive lesions. Ground-glass opacities, cavitating nodules/masses and masses measuring more than 3 cm represent active disease ordinarily. Non-cavitary small nodules and septal or non-septal lines can be either active or cicatricial lesions. The nature of these lesions needs to be clarified by longitudinal observation. (orig.)

  3. Influência do período de restrição hídrica na atividade residual de isoxaflutole no solo Influence of drought periods on the residual activity of isoxaflutole in soil

    Directory of Open Access Journals (Sweden)

    R.S. Oliveira Jr.

    2006-12-01

    Full Text Available O objetivo deste trabalho foi estudar a atividade residual do isoxaflutole (IFT no controle de Brachiaria decumbens sob diferentes períodos de seca, após a aplicação do herbicida, em dois solos. Para isso, foram realizados seis ensaios simultâneos, sendo utilizadas doses de 0, 230 e 270 g ha-1 de isoxaflutole em amostras de Latossolo Vermelho Distroférrico nitossólico (textura argilosa e doses de 0, 180 e 200 g ha-1 em amostras de Latossolo Vermelho distrófico (textura franco-argilo-arenosa. Em cada ensaio, foram combinados em esquema fatorial doses e períodos de seca após as aplicações do herbicida, utilizando-se B. decumbens como bioindicador da atividade residual do herbicida no solo. As avaliações de controle foram feitas aos 15, 30, 45 e 60 dias após a semeadura do bioindicador. O IFT apresentou alta estabilidade no solo de textura argilosa mesmo após três chuvas simuladas de 20 mm, espaçadas de 30 dias e seguidas de 120 dias de seca após sua aplicação. No solo de textura franco-arenosa a estabilidade foi menor, e o efeito residual (80% de controle persistiu entre 0 e 110 dias para B. decumbens, conforme dose, períodos de seca e data de avaliação após a semeadura. Esse fato evidenciou que, à medida que aumenta o tempo e o número de irrigações entre a aplicação do herbicida e a semeadura do bioindicador, há redução no potencial efetivo de controle de IFT no Latossolo Vermelho distrófico.This work was carried out to evaluate the residual activity of isoxaflutole (IFT in Brachiaria decumbens control after different simulated drought conditions following herbicide application in two soils. Six simultaneous experiments were performed, using rates of 0, 230 and 270 g ha-1 of IFT in samples of a heavy clay soil and of 0, 180 and 200 g ha-1 in samples of a sandy clay loam soil. In each experiment, rates of IFT were combined in a factorial scheme with drought periods after herbicide application. B. decumbens was

  4. Biological activities and molecular interactions of the C-terminal residue of thrombospondin-4, an epitome of acidic amphipathic peptides.

    Science.gov (United States)

    Congote, Luis F; Sadvakassova, Gulzhakhan; Dobocan, Monica C; Difalco, Marcos R; Kriazhev, Leonid

    2010-04-01

    C21, the C-terminal residue of thrombospondin-4 (TSP-4), was identified as a peptide growth factor during an investigation concerning erythropoietin-dependent, erythroid stimulating factors of endothelial origin. It is active in cultures of several human hematopoietic stem cells, skin fibroblasts and kidney epithelial cells and stimulates red cell formation in anemic mice. A method of affinity chromatography in the presence of high concentrations of Triton X-100, previously developed for identifying proteins associated with the TSP-1 receptor CD47, was utilized for the detection of C21 binding molecules and their detergent-resistant, associated partners. These experiments helped to delineate two different mechanisms of C21 action, which are compatible with its cell proliferating activity. As a cell matrix peptide, C21 binds to the osteopontin receptor CD44 and could act as an osteopontin antagonist, preventing the inhibition of primitive hematopoietic stem cell proliferation. TSP-1, another matrix protein, binds to C21 and could indirectly act as an antagonist, by shunting C21-CD44 interactions. The second mechanism is a direct effect of C21 on cell proliferation. The extremely rapid internalization and nuclear localization of the peptide could be explained by CD44-mediated internalization, followed by a microtubule-mediated transport towards the nucleus, or, eventually, direct membrane insertion. These alternative hypotheses are supported by previously observed membrane insertion of similar synthetic and viral acidic amphipathic peptides, the presence of microtubule-associated protein 1B (MAP1B) and dynactin in the triton-soluble complexes associated with C21 and the presence in such complexes of dual compartment proteins for nuclei and plasma membranes, such as MAP1B, AHNAK and CD44. Copyright (c) 2009 Elsevier Inc. All rights reserved.

  5. Residue processing

    Energy Technology Data Exchange (ETDEWEB)

    Gieg, W.; Rank, V.

    1942-10-15

    In the first stage of coal hydrogenation, the liquid phase, light and heavy oils were produced; the latter containing the nonliquefied parts of the coal, the coal ash, and the catalyst substances. It was the problem of residue processing to extract from these so-called let-down oils that which could be used as pasting oils for the coal. The object was to obtain a maximum oil extraction and a complete removal of the solids, because of the latter were returned to the process they would needlessly burden the reaction space. Separation of solids in residue processing could be accomplished by filtration, centrifugation, extraction, distillation, or low-temperature carbonization (L.T.C.). Filtration or centrifugation was most suitable since a maximum oil yield could be expected from it, since only a small portion of the let-down oil contained in the filtration or centrifugation residue had to be thermally treated. The most satisfactory centrifuge at this time was the Laval, which delivered liquid centrifuge residue and centrifuge oil continuously. By comparison, the semi-continuous centrifuges delivered plastic residues which were difficult to handle. Various apparatus such as the spiral screw kiln and the ball kiln were used for low-temperature carbonization of centrifuge residues. Both were based on the idea of carbonization in thin layers. Efforts were also being made to produce electrode carbon and briquette binder as by-products of the liquid coal phase.

  6. A stretch of polybasic residues mediates Cdc42 GTPase-activating protein (CdGAP) binding to phosphatidylinositol 3,4,5-trisphosphate and regulates its GAP activity.

    Science.gov (United States)

    Karimzadeh, Fereshteh; Primeau, Martin; Mountassif, Driss; Rouiller, Isabelle; Lamarche-Vane, Nathalie

    2012-06-01

    The Rho family of small GTPases are membrane-associated molecular switches involved in the control of a wide range of cellular activities, including cell migration, adhesion, and proliferation. Cdc42 GTPase-activating protein (CdGAP) is a phosphoprotein showing GAP activity toward Rac1 and Cdc42. CdGAP activity is regulated in an adhesion-dependent manner and more recently, we have identified CdGAP as a novel molecular target in signaling and an essential component in the synergistic interaction between TGFβ and Neu/ErbB-2 signaling pathways in breast cancer cells. In this study, we identified a small polybasic region (PBR) preceding the RhoGAP domain that mediates specific binding to negatively charged phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3). In vitro reconstitution of membrane vesicles loaded with prenylated Rac1 demonstrates that the PBR is required for full activation of CdGAP in the presence of PI(3,4,5)P3. In fibroblast cells, the expression of CdGAP protein mutants lacking an intact PBR shows a significant reduced ability of the protein mutants to induce cell rounding or to mediate negative effects on cell spreading. Furthermore, an intact PBR is required for CdGAP to inactivate Rac1 signaling into cells, whereas it is not essential in an in vitro context. Altogether, these studies reveal that specific interaction between negatively charged phospholipid PI(3,4,5)P3 and the stretch of polybasic residues preceding the RhoGAP domain regulates CdGAP activity in vivo and is required for its cellular functions.

  7. A small-molecule inhibitor of the ubiquitin activating enzyme for cancer treatment.

    Science.gov (United States)

    Hyer, Marc L; Milhollen, Michael A; Ciavarri, Jeff; Fleming, Paul; Traore, Tary; Sappal, Darshan; Huck, Jessica; Shi, Judy; Gavin, James; Brownell, Jim; Yang, Yu; Stringer, Bradley; Griffin, Robert; Bruzzese, Frank; Soucy, Teresa; Duffy, Jennifer; Rabino, Claudia; Riceberg, Jessica; Hoar, Kara; Lublinsky, Anya; Menon, Saurabh; Sintchak, Michael; Bump, Nancy; Pulukuri, Sai M; Langston, Steve; Tirrell, Stephen; Kuranda, Mike; Veiby, Petter; Newcomb, John; Li, Ping; Wu, Jing Tao; Powe, Josh; Dick, Lawrence R; Greenspan, Paul; Galvin, Katherine; Manfredi, Mark; Claiborne, Chris; Amidon, Benjamin S; Bence, Neil F

    2018-02-01

    The ubiquitin-proteasome system (UPS) comprises a network of enzymes that is responsible for maintaining cellular protein homeostasis. The therapeutic potential of this pathway has been validated by the clinical successes of a number of UPS modulators, including proteasome inhibitors and immunomodulatory imide drugs (IMiDs). Here we identified TAK-243 (formerly known as MLN7243) as a potent, mechanism-based small-molecule inhibitor of the ubiquitin activating enzyme (UAE), the primary mammalian E1 enzyme that regulates the ubiquitin conjugation cascade. TAK-243 treatment caused depletion of cellular ubiquitin conjugates, resulting in disruption of signaling events, induction of proteotoxic stress, and impairment of cell cycle progression and DNA damage repair pathways. TAK-243 treatment caused death of cancer cells and, in primary human xenograft studies, demonstrated antitumor activity at tolerated doses. Due to its specificity and potency, TAK-243 allows for interrogation of ubiquitin biology and for assessment of UAE inhibition as a new approach for cancer treatment.

  8. A Conserved Active Site Tyrosine Residue of Proline Dehydrogenase Helps Enforce the Preference for Proline over Hydroxyproline as the Substrate

    Energy Technology Data Exchange (ETDEWEB)

    Ostrander, E.L.; Larson, J.D.; Schuermann, J.P.; Tanner, J.J.; (Cornell); (Missouri)

    2009-03-02

    Proline dehydrogenase (PRODH) catalyzes the oxidation of L-proline to {Delta}-1-pyrroline-5-carboxylate. PRODHs exhibit a pronounced preference for proline over hydroxyproline (trans-4-hydroxy-L-proline) as the substrate, but the basis for specificity is unknown. The goal of this study, therefore, is to gain insight into the structural determinants of substrate specificity of this class of enzyme, with a focus on understanding how PRODHs discriminate between the two closely related molecules, proline and hydroxyproline. Two site-directed mutants of the PRODH domain of Escherichia coli PutA were created: Y540A and Y540S. Kinetics measurements were performed with both mutants. Crystal structures of Y540S complexed with hydroxyproline, proline, and the proline analogue L-tetrahydro-2-furoic acid were determined at resolutions of 1.75, 1.90, and 1.85 {angstrom}, respectively. Mutation of Tyr540 increases the catalytic efficiency for hydroxyproline 3-fold and decreases the specificity for proline by factors of 20 (Y540S) and 50 (Y540A). The structures show that removal of the large phenol side chain increases the volume of the substrate-binding pocket, allowing sufficient room for the 4-hydroxyl of hydroxyproline. Furthermore, the introduced serine residue participates in recognition of hydroxyproline by forming a hydrogen bond with the 4-hydroxyl. This result has implications for understanding the substrate specificity of the related enzyme human hydroxyproline dehydrogenase, which has serine in place of tyrosine at this key active site position. The kinetic and structural results suggest that Tyr540 is an important determinant of specificity. Structurally, it serves as a negative filter for hydroxyproline by clashing with the 4-hydroxyl group of this potential substrate.

  9. Residues Phe103 and Phe149 are critical for the co-chaperone activity of Bacillus licheniformis GrpE.

    Science.gov (United States)

    Lin, Min-Guan; Chi, Meng-Chun; Chen, Bo-En; Wang, Tzu-Fan; Lo, Huei-Fen; Lin, Long-Liu

    2015-01-01

    A tryptophan-free Bacillus licheniformis nucleotide exchange factor (BlGrpE) and its Trp mutants (F70W, F103W, F149W, F70/103W, F70/149W, F103/149W and F70/103/149W) were over-expressed and purified to near homogeneity. Simultaneous addition of B. licheniformis DnaJ, NR-peptide and individual variants synergistically stimulated the ATPase activity of a recombinant DnaK (BlDnaK) from the same bacterium by 3.1-14.7-fold, which are significantly lower than the synergistic stimulation (18.9-fold) of BlGrpE. Protein-protein interaction analysis revealed that Trp mutants relevant to amino acid positions 103 and 149 lost the ability to bind BlDnaK. Circular dichroism measurements indicate that F70W displayed a comparable level of secondary structure to that of BlGrpE, and the wild-type protein and the Trp mutants as well all experienced a reversible behavior of thermal denaturation. Guanidine hydrochloride (GdnHCl)-induced unfolding transition for BlGrpE was calculated to be 1.25 M corresponding to ΔG(N-U) of 4.29 kcal/mol, whereas the unfolding transitions of mutant proteins were in the range of 0.77-1.31 M equivalent to ΔG(N-U) of 2.41-4.14 kcal/mol. Taken together, the introduction of tryptophan residue, especially at positions 103 and 149, into the primary structure of BlGrpE has been proven to be detrimental to structural integrity and proper function of the protein. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Efficacy and safety of TAK-085 compared with eicosapentaenoic acid in Japanese subjects with hypertriglyceridemia undergoing lifestyle modification: the omega-3 fatty acids randomized double-blind (ORD) study.

    Science.gov (United States)

    Tatsuno, Ichiro; Saito, Yasushi; Kudou, Kentarou; Ootake, Jun

    2013-01-01

    Hypertriglyceridemia is a risk factor for cardiovascular disease, and clinical practice guidelines advocate treatment to reduce triglyceride (TG) levels. In Japan, an EPA-E (eicosapentaenoic acid-ethyl ester) product has been used clinically for treating dyslipidemia. We investigated the TG-lowering effects of TAK-085 (EPA-E + docosahexaenoic acid-ethyl ester) in comparison with EPA-E in Japanese patients with hypertriglyceridemia (TG ≥150 mg/dL and acid in TAK-085 contains approximately 465 mg of EPA plus 375 mg of docosahexaenoic acid-ethyl as ethyl esters. Guidance on lifestyle modifications was provided throughout. The primary end point was the percent change in TG levels (baseline from end of treatment), which was -10.8 ± 22.6, -22.9 ± 23.1, and -11.2 ± 25.7 in the TAK-085 2 g/day, TAK-085 4 g/day, and EPA-E 1.8 g/day groups, respectively. TAK-085 4 g/day produced a significantly greater reduction in TG than EPA-E 1.8 g/day (P < .0001), whereas TAK-085 2 g/day was not inferior to EPA-E 1.8 g/day. Changes in other lipid parameters were relatively modest. There were no notable safety or tolerability differences between the groups. In Japanese patients with modest hypertriglyceridemia who also underwent lifestyle intervention, TAK-085 4 g/day reduced TG more than EPA-E 1.8 g/day. TAK-085 2 g/day had similar effects on TG as EPA-E 1.8 g/day. TAK-085 was well-tolerated. Copyright © 2013 National Lipid Association. Published by Elsevier Inc. All rights reserved.

  11. Phytochemical Composition, Antioxidant Activity, and the Effect of the Aqueous Extract of Coffee (Coffea arabica L. Bean Residual Press Cake on the Skin Wound Healing

    Directory of Open Access Journals (Sweden)

    Regina Celis Lopes Affonso

    2016-01-01

    Full Text Available The world coffee consumption has been growing for its appreciated taste and its beneficial effects on health. The residual biomass of coffee, originated in the food industry after oil extraction from coffee beans, called coffee beans residual press cake, has attracted interest as a source of compounds with antioxidant activity. This study investigated the chemical composition of aqueous extracts of coffee beans residual press cake (AE, their antioxidant activity, and the effect of topical application on the skin wound healing, in animal model, of hydrogels containing the AE, chlorogenic acid (CGA, allantoin (positive control, and carbopol (negative control. The treatments’ performance was compared by measuring the reduction of the wound area, with superior result (p<0.05 for the green coffee AE (78.20% with respect to roasted coffee AE (53.71%, allantoin (70.83%, and carbopol (23.56%. CGA hydrogels reduced significantly the wound area size on the inflammatory phase, which may be associated with the well known antioxidant and anti-inflammatory actions of that compound. The topic use of the coffee AE studied improved the skin wound healing and points to an interesting biotechnological application of the coffee bean residual press cake.

  12. Phytochemical Composition, Antioxidant Activity, and the Effect of the Aqueous Extract of Coffee (Coffea arabicaL.) Bean Residual Press Cake on the Skin Wound Healing.

    Science.gov (United States)

    Affonso, Regina Celis Lopes; Voytena, Ana Paula Lorenzen; Fanan, Simone; Pitz, Heloísa; Coelho, Daniela Sousa; Horstmann, Ana Luiza; Pereira, Aline; Uarrota, Virgílio Gavicho; Hillmann, Maria Clara; Varela, Lucas Andre Calbusch; Ribeiro-do-Valle, Rosa Maria; Maraschin, Marcelo

    2016-01-01

    The world coffee consumption has been growing for its appreciated taste and its beneficial effects on health. The residual biomass of coffee, originated in the food industry after oil extraction from coffee beans, called coffee beans residual press cake, has attracted interest as a source of compounds with antioxidant activity. This study investigated the chemical composition of aqueous extracts of coffee beans residual press cake (AE), their antioxidant activity, and the effect of topical application on the skin wound healing, in animal model, of hydrogels containing the AE, chlorogenic acid (CGA), allantoin (positive control), and carbopol (negative control). The treatments' performance was compared by measuring the reduction of the wound area, with superior result ( p coffee AE (78.20%) with respect to roasted coffee AE (53.71%), allantoin (70.83%), and carbopol (23.56%). CGA hydrogels reduced significantly the wound area size on the inflammatory phase, which may be associated with the well known antioxidant and anti-inflammatory actions of that compound. The topic use of the coffee AE studied improved the skin wound healing and points to an interesting biotechnological application of the coffee bean residual press cake.

  13. Handling of Solid Residues

    International Nuclear Information System (INIS)

    Medina Bermudez, Clara Ines

    1999-01-01

    The topic of solid residues is specifically of great interest and concern for the authorities, institutions and community that identify in them a true threat against the human health and the atmosphere in the related with the aesthetic deterioration of the urban centers and of the natural landscape; in the proliferation of vectorial transmitters of illnesses and the effect on the biodiversity. Inside the wide spectrum of topics that they keep relationship with the environmental protection, the inadequate handling of solid residues and residues dangerous squatter an important line in the definition of political and practical environmentally sustainable. The industrial development and the population's growth have originated a continuous increase in the production of solid residues; of equal it forms, their composition day after day is more heterogeneous. The base for the good handling includes the appropriate intervention of the different stages of an integral administration of residues, which include the separation in the source, the gathering, the handling, the use, treatment, final disposition and the institutional organization of the administration. The topic of the dangerous residues generates more expectation. These residues understand from those of pathogen type that are generated in the establishments of health that of hospital attention, until those of combustible, inflammable type, explosive, radio-active, volatile, corrosive, reagent or toxic, associated to numerous industrial processes, common in our countries in development

  14. Preclinical FLT-PET and FDG-PET imaging of tumor response to the multi-targeted Aurora B kinase inhibitor, TAK-901

    International Nuclear Information System (INIS)

    Cullinane, Carleen; Waldeck, Kelly L.; Binns, David; Bogatyreva, Ekaterina; Bradley, Daniel P.; Jong, Ron de; McArthur, Grant A.; Hicks, Rodney J.

    2014-01-01

    Introduction: The Aurora kinases play a key role in mitosis and have recently been identified as attractive targets for therapeutic intervention in cancer. The aim of this study was therefore to investigate the utility of 3′-[ 18 F]fluoro-3′-deoxythymidine (FLT) and 2-deoxy-2-[ 18 F]fluoro-D-glucose (FDG) for assessment of tumor response to the multi-targeted Aurora B kinase inhibitor, TAK-901. Methods: Balb/c nude mice bearing HCT116 colorectal xenografts were treated with up to 30 mg/kg TAK 901 or vehicle intravenously twice daily for two days on a weekly cycle. Tumor growth was monitored by calliper measurements and PET imaging was performed at baseline, day 4, 8, 11 and 15. Tumors were harvested at time points corresponding to days of PET imaging for analysis of ex vivo markers of cell proliferation and metabolism together with markers of Aurora B kinase inhibition including phospho-histone H3 (pHH3) and senescence associated β-galactosidase. Results: Tumor growth was inhibited by 60% on day 12 of 30 mg/kg TAK-901 therapy. FLT uptake was significantly reduced by day 4 of treatment and this corresponded with reduction in bromodeoxyuridine and pHH3 staining by immunohistochemistry. All biomarkers rebounded towards baseline levels by the commencement of the next treatment cycle, consistent with release of Aurora B kinase suppression. TAK-901 therapy had no impact on glucose metabolism as assessed by FDG uptake and GLUT1 staining by immunohistochemistry. Conclusions: FLT-PET, but not FDG-PET, is a robust non-invasive imaging biomarker of early HCT116 tumor response to the on-target effects of the multi-targeted Aurora B kinase inhibitor, TAK-901. Advances in knowledge and implications for patient care: This is the first report to demonstrate the impact of the multi-targeted Aurora B kinase inhibitor, TAK-901 on tumor FLT uptake. The findings provide a strong rationale for the evaluation of FLT-PET as an early biomarker of tumor response in the early phase

  15. ACR/EULAR Definitions of Remission Are Associated with Lower Residual Inflammatory Activity Compared with DAS28 Remission on Hand MRI in Rheumatoid Arthritis.

    Science.gov (United States)

    Lisbona, Maria Pilar; Solano, Albert; Ares, Jesús; Almirall, Miriam; Salman-Monte, Tarek Carlos; Maymó, Joan

    2016-09-01

    To determine the level of residual inflammation [synovitis, bone marrow edema (BME), tenosynovitis, and total inflammation] quantified by hand magnetic resonance imaging (h-MRI) in patients with rheumatoid arthritis (RA) in remission according to 3 different definitions of clinical remission, and to compare these remission definitions. A cross-sectional study. To assess the level of residual MRI inflammation in remission, cutoff levels associated to remission and median scores of MRI residual inflammatory lesions were calculated. Data from an MRI register of patients with RA who have various levels of disease activity were used. These were used for the analyses: synovitis, BME according to the Rheumatoid Arthritis Magnetic Resonance Imaging Scoring system, tenosynovitis, total inflammation, and disease activity composite measures recorded at the time of MRI. Receiver-operating characteristic analysis was used to identify the best cutoffs associated with remission for each inflammatory lesion on h-MRI. Median values of each inflammatory lesion for each definition of remission were also calculated. A total of 388 h-MRI sets of patients with RA with different levels of disease activity, 130 in remission, were included. Cutoff values associated with remission according to the Simplified Disease Activity Index (SDAI) ≤ 3.3 and the Boolean American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) definitions for BME and tenosynovitis (1 and 3, respectively) were lower than BME and tenosynovitis (2 and 5, respectively) for the Disease Activity Score on 28 joints (DAS28) ≤ 2.6. Median scores for synovitis, BME, and total inflammation were also lower for the SDAI and Boolean ACR/EULAR remission criteria compared with DAS28. Patients with RA in remission according to the SDAI and Boolean ACR/EULAR definitions showed lower levels of MRI-detected residual inflammation compared with DAS28.

  16. Structural Characterization and Antioxidative Activity of Low-Molecular-Weights Beta-1,3-Glucan from the Residue of Extracted Ganoderma lucidum Fruiting Bodies

    Directory of Open Access Journals (Sweden)

    Pai-Feng Kao

    2012-01-01

    Full Text Available The major cell wall constituent of Ganoderma lucidum (G. lucidum is β-1,3-glucan. This study examined the polysaccharide from the residues of alkaline-extracted fruiting bodies using high-performance anion-exchange chromatography (HPAEC, and it employed nuclear magnetic resonance (NMR and mass spectrometry (MS to confirm the structures. We have successfully isolated low-molecular-weight β-1,3-glucan (LMG, in high yields, from the waste residue of extracted fruiting bodies of G. lucidum. The 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT assay evaluated the capability of LMG to suppress H2O2-induced cell death in RAW264.7 cells, identifying that LMG protected cells from H2O2-induced damage. LMG treatment decreased H2O2-induced intracellular reactive oxygen species (ROS production. LMG also influenced sphingomyelinase (SMase activity, stimulated by cell death to induce ceramide formation, and then increase cell ROS production. Estimation of the activities of neutral and acid SMases in vitro showed that LMG suppressed the activities of both neutral and acid SMases in a concentration-dependent manner. These results suggest that LMG, a water-soluble β-1,3-glucan recycled from extracted residue of G. lucidum, possesses antioxidant capability against H2O2-induced cell death by attenuating intracellular ROS and inhibiting SMase activity.

  17. Residual risk

    African Journals Online (AJOL)

    ing the residual risk of transmission of HIV by blood transfusion. An epidemiological approach assumed that all HIV infections detected serologically in first-time donors were pre-existing or prevalent infections, and that all infections detected in repeat blood donors were new or incident infections. During 1986 - 1987,0,012%.

  18. Influence of amino acid residues near the active site of cytochrome P450 from Bacillus megaterium on the selectivity of n-octane oxidation to octanol regioisomers

    Science.gov (United States)

    Miyaji, Akimitsu; Baba, Toshihide

    2017-09-01

    A mutant of cytochrome P450 from Bacillus megaterium (CYP450BM-3) was prepared by replacing two alanine residues around active site of the enzyme, alanine 328 and alanine 82, with leucine and tryptophan, respectively. The CYP450BM-3 mutant produced 2-octanol selectively from n-octane under atmospheric temperature and pressure; its selectivity was 74%. Furthermore, the mutant produced 1-octanol, which is not produced by wild-type enzyme.

  19. Residues within the Transmembrane Domain of the Glucagon-Like Peptide-1 Receptor Involved in Ligand Binding and Receptor Activation: Modelling the Ligand-Bound Receptor

    Science.gov (United States)

    Coopman, K.; Wallis, R.; Robb, G.; Brown, A. J. H.; Wilkinson, G. F.; Timms, D.

    2011-01-01

    The C-terminal regions of glucagon-like peptide-1 (GLP-1) bind to the N terminus of the GLP-1 receptor (GLP-1R), facilitating interaction of the ligand N terminus with the receptor transmembrane domain. In contrast, the agonist exendin-4 relies less on the transmembrane domain, and truncated antagonist analogs (e.g. exendin 9–39) may interact solely with the receptor N terminus. Here we used mutagenesis to explore the role of residues highly conserved in the predicted transmembrane helices of mammalian GLP-1Rs and conserved in family B G protein coupled receptors in ligand binding and GLP-1R activation. By iteration using information from the mutagenesis, along with the available crystal structure of the receptor N terminus and a model of the active opsin transmembrane domain, we developed a structural receptor model with GLP-1 bound and used this to better understand consequences of mutations. Mutation at Y152 [transmembrane helix (TM) 1], R190 (TM2), Y235 (TM3), H363 (TM6), and E364 (TM6) produced similar reductions in affinity for GLP-1 and exendin 9–39. In contrast, other mutations either preferentially [K197 (TM2), Q234 (TM3), and W284 (extracellular loop 2)] or solely [D198 (TM2) and R310 (TM5)] reduced GLP-1 affinity. Reduced agonist affinity was always associated with reduced potency. However, reductions in potency exceeded reductions in agonist affinity for K197A, W284A, and R310A, while H363A was uncoupled from cAMP generation, highlighting critical roles of these residues in translating binding to activation. Data show important roles in ligand binding and receptor activation of conserved residues within the transmembrane domain of the GLP-1R. The receptor structural model provides insight into the roles of these residues. PMID:21868452

  20. The E-screen test and the MELN gene-reporter assay used for determination of estrogenic activity in fruits and vegetables in relation to pesticide residues.

    Science.gov (United States)

    Schilirò, Tiziana; Porfido, Arianna; Longo, Annalisa; Coluccia, Sara; Gilli, Giorgio

    2013-12-01

    Endocrine-disrupting chemicals (EDCs) may lead to adverse systemic effects by interfering with normal hormone homeostasis, and diet is considered to be among the main routes of EDC exposure. The present study investigated the total estrogenic activity of fruits and vegetables by calculating the 17-β-estradiol equivalent quantity (EEQ) using two in vitro tests: the human breast cancer cell line (MCF-7 BUS) proliferation assay (E-screen test) and the luciferase-transfected human breast cancer cell line (MELN) gene-reporter assay. Of the 24 analyzed fruits and vegetables, 14 contained from 1 to 4 pesticide residues in concentrations ranging from 0.02 to 1.19 ppm, whereas the other 10 did not contain any pesticide residues. The EEQ values for all positive samples ranged from 0.010 to 0.616 μg/100g for the above in vitro tests. Our study demonstrates that estrogenic activity was present in fruits and vegetables and that the concentration of allowable pesticide residues and EEQ values were positively correlated; however, no correlation was found by comparing the estrogenic activity and the intrinsic content of phytoestrogens obtained from the available literature. A theoretical adult dietary intake of 0.7-0.9 ng EEQ/L/day from fruits and vegetables was calculated. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. Management of NORM Residues

    International Nuclear Information System (INIS)

    2013-06-01

    The IAEA attaches great importance to the dissemination of information that can assist Member States in the development, implementation, maintenance and continuous improvement of systems, programmes and activities that support the nuclear fuel cycle and nuclear applications, and that address the legacy of past practices and accidents. However, radioactive residues are found not only in nuclear fuel cycle activities, but also in a range of other industrial activities, including: - Mining and milling of metalliferous and non-metallic ores; - Production of non-nuclear fuels, including coal, oil and gas; - Extraction and purification of water (e.g. in the generation of geothermal energy, as drinking and industrial process water; in paper and pulp manufacturing processes); - Production of industrial minerals, including phosphate, clay and building materials; - Use of radionuclides, such as thorium, for properties other than their radioactivity. Naturally occurring radioactive material (NORM) may lead to exposures at some stage of these processes and in the use or reuse of products, residues or wastes. Several IAEA publications address NORM issues with a special focus on some of the more relevant industrial operations. This publication attempts to provide guidance on managing residues arising from different NORM type industries, and on pertinent residue management strategies and technologies, to help Member States gain perspectives on the management of NORM residues

  2. Authorization and Toxicity of Veterinary Drugs and Plant Protection Products: Residues of the Active Ingredients in Food and Feed and Toxicity Problems Related to Adjuvants

    Directory of Open Access Journals (Sweden)

    Szandra Klátyik

    2017-09-01

    Full Text Available Chemical substances applied in animal husbandry or veterinary medicine and in crop protection represent substantial environmental loads, and their residues occur in food and feed products. Product approval is governed differently in these two sectors in the European Union (EU, and the occurrence of veterinary drug (VD and pesticide residues indicated by contamination notification cases in the Rapid Alert System for Food and Feed of the EU also show characteristic differences. While the initial high numbers of VD residues reported in 2002 were successfully suppressed to less than 100 cases annually by 2006 and on, the number of notification cases for pesticide residues showed a gradual increase from a low (approximately 50 cases annually initial level until 2005 to more than 250 cases annually after 2009, with a halt occurring only in 2016. Main notifiers of VD residues include Germany, Belgium, the UK, and Italy (63, 59, 42, and 31 notifications announced, respectively, and main consigning countries of non-compliances are Vietnam, India, China, and Brazil (88, 50, 34, and 23 notifications, respectively. Thus, countries of South and Southeast Asia are considered a vulnerable point with regard to VD residues entering the EU market. Unintended side effects of VDs and plant protection products may be caused not only by the active ingredients but also by various additives in these preparations. Adjuvants (e.g., surfactants and other co-formulants used in therapeutic agents and feed additives, as well as in pesticide formulations have long been considered as inactive ingredients in the aspects of the required main biological effect of the pharmaceutical or pesticide, and in turn, legal regulations of the approval and marketing of these additives specified significantly less stringent risk assessment requirements, than those specified for the active ingredients. However, numerous studies have shown additive, synergistic, or antagonistic side effects

  3. Authorization and Toxicity of Veterinary Drugs and Plant Protection Products: Residues of the Active Ingredients in Food and Feed and Toxicity Problems Related to Adjuvants.

    Science.gov (United States)

    Klátyik, Szandra; Bohus, Péter; Darvas, Béla; Székács, András

    2017-01-01

    Chemical substances applied in animal husbandry or veterinary medicine and in crop protection represent substantial environmental loads, and their residues occur in food and feed products. Product approval is governed differently in these two sectors in the European Union (EU), and the occurrence of veterinary drug (VD) and pesticide residues indicated by contamination notification cases in the Rapid Alert System for Food and Feed of the EU also show characteristic differences. While the initial high numbers of VD residues reported in 2002 were successfully suppressed to less than 100 cases annually by 2006 and on, the number of notification cases for pesticide residues showed a gradual increase from a low (approximately 50 cases annually) initial level until 2005 to more than 250 cases annually after 2009, with a halt occurring only in 2016. Main notifiers of VD residues include Germany, Belgium, the UK, and Italy (63, 59, 42, and 31 notifications announced, respectively), and main consigning countries of non-compliances are Vietnam, India, China, and Brazil (88, 50, 34, and 23 notifications, respectively). Thus, countries of South and Southeast Asia are considered a vulnerable point with regard to VD residues entering the EU market. Unintended side effects of VDs and plant protection products may be caused not only by the active ingredients but also by various additives in these preparations. Adjuvants (e.g., surfactants) and other co-formulants used in therapeutic agents and feed additives, as well as in pesticide formulations have long been considered as inactive ingredients in the aspects of the required main biological effect of the pharmaceutical or pesticide, and in turn, legal regulations of the approval and marketing of these additives specified significantly less stringent risk assessment requirements, than those specified for the active ingredients. However, numerous studies have shown additive, synergistic, or antagonistic side effects between the

  4. Authorization and Toxicity of Veterinary Drugs and Plant Protection Products: Residues of the Active Ingredients in Food and Feed and Toxicity Problems Related to Adjuvants

    Science.gov (United States)

    Klátyik, Szandra; Bohus, Péter; Darvas, Béla; Székács, András

    2017-01-01

    Chemical substances applied in animal husbandry or veterinary medicine and in crop protection represent substantial environmental loads, and their residues occur in food and feed products. Product approval is governed differently in these two sectors in the European Union (EU), and the occurrence of veterinary drug (VD) and pesticide residues indicated by contamination notification cases in the Rapid Alert System for Food and Feed of the EU also show characteristic differences. While the initial high numbers of VD residues reported in 2002 were successfully suppressed to less than 100 cases annually by 2006 and on, the number of notification cases for pesticide residues showed a gradual increase from a low (approximately 50 cases annually) initial level until 2005 to more than 250 cases annually after 2009, with a halt occurring only in 2016. Main notifiers of VD residues include Germany, Belgium, the UK, and Italy (63, 59, 42, and 31 notifications announced, respectively), and main consigning countries of non-compliances are Vietnam, India, China, and Brazil (88, 50, 34, and 23 notifications, respectively). Thus, countries of South and Southeast Asia are considered a vulnerable point with regard to VD residues entering the EU market. Unintended side effects of VDs and plant protection products may be caused not only by the active ingredients but also by various additives in these preparations. Adjuvants (e.g., surfactants) and other co-formulants used in therapeutic agents and feed additives, as well as in pesticide formulations have long been considered as inactive ingredients in the aspects of the required main biological effect of the pharmaceutical or pesticide, and in turn, legal regulations of the approval and marketing of these additives specified significantly less stringent risk assessment requirements, than those specified for the active ingredients. However, numerous studies have shown additive, synergistic, or antagonistic side effects between the

  5. Pengaruh Pemanasan Awal dan Medan Elektromagnet pada Biodiesel terhadap Daya dan Kepekatan Gas Buang pada Motor Diesel 4 Tak 4 Silinder

    OpenAIRE

    soares, Jacinto; Sahbana, Muhammad Agus; Fuhaid, Naif

    2013-01-01

    Salah satu permasalahan di bidang otomotif adalah ketersediaan bahan bakar fosil minyak bumi. Bahan bakar Bio menjadi salah satu sumber alternatif untuk mesin diesel. Bahan bakar tersebut memerlukan perlakuan tertentu sebelum memasuki ruang bakar di mesin. Penelitian ini bertujuan untuk mendapatkan pengaruh pemanasan awal dan medan elektromagnet pada biodiesel terhadap daya dan kepekatan gas buang motor diesel 4 tak 4 silinder. Eksperimen dilakukan mengunakan mobil chevrolet/isuzu. Bahan baka...

  6. Plasminogen activator inhibitor-1 4G/5G genotype and residual venous occlusion following acute unprovoked deep vein thrombosis of the lower limb: A prospective cohort study.

    Science.gov (United States)

    Giurgea, Georgiana-Aura; Brunner-Ziegler, Sophie; Jilma, Bernd; Sunder-Plassmann, Raute; Koppensteiner, Renate; Gremmel, Thomas

    2017-05-01

    A recent study suggested that the plasminogen activator inhibitor (PAI)-1 4G/5G genotype may play a role in the resolution of deep vein thrombosis (DVT) after surgery. In the present study, we investigated the association between PAI-1 4G/5G genotype and the persistence of venous occlusion after acute idiopathic DVT of the lower limb. The PAI-1 4G/5G genotype was determined by real-Time PCR in 43 patients with unprovoked DVT of the lower limb. Residual venous occlusion was assessed by duplex sonography 1, 3, 6, 12 and 24months after the acute event. The PAI-1 Activity was determined by ELISA. Ten patients (23%) were homozygous for 4G (4G/4G), 27 patients (63%) were heterozygous 4G/5G and 6 patients (14%) were homozygous for 5G (5G/5G). Residual venous occlusion (RVO) was found in 77%, 65%, 58%, 56% and 37% of the overall study population, at 1, 3, 6, 12 and 24months after acute DVT, respectively. The presence of residual venous occlusion at 1, 3, 6, 12 and 24months after acute unprovoked DVT did not differ significantly between genotypes, but age was associated with RVO. Plasma levels of PAI-1 activity correlated with body mass index but was not associated with genotypes in our study. The PAI-1 4G/5G genotype was not a relevant predictor of persistent residual venous occlusion after idiopathic DVT, which however was associated with age. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Residual basins

    International Nuclear Information System (INIS)

    D'Elboux, C.V.; Paiva, I.B.

    1980-01-01

    Exploration for uranium carried out over a major portion of the Rio Grande do Sul Shield has revealed a number of small residual basins developed along glacially eroded channels of pre-Permian age. Mineralization of uranium occurs in two distinct sedimentary units. The lower unit consists of rhythmites overlain by a sequence of black shales, siltstones and coal seams, while the upper one is dominated by sandstones of probable fluvial origin. (Author) [pt

  8. Atividade residual de imazethapyr + imazapic em arroz semeado em rotação com o arroz Clearfield® Residual soil activity of imazethapyr + imazapic to rice planted in rotation after Clearfield rice®

    Directory of Open Access Journals (Sweden)

    J.J.O. Pinto

    2011-03-01

    Full Text Available Objetivou-se com este trabalho avaliar os efeitos da atividade residual da mistura comercial do herbicida Only® (imazethapyr+imazapic sobre o arroz convencional, cultivar IRGA 417, em solo cultivado em uma e três safras sequenciais de arroz Clearfield® (CL. Quatro experimentos foram conduzidos no município de Capão do Leão, RS, em delineamento de blocos ao acaso, com quatro e oito repetições, respectivamente para dois experimentos de campo (EC1 e EC3, e dois em casa de vegetação (CV1 e CV3. Os tratamentos foram arranjados em esquema fatorial; o fator A, dentro da mesma safra, comparava resíduos em repetições de anos de cultivo, e o fator B avaliava a dose aplicada sobre o arroz CL e a atividade residual do herbicida ao arroz não mutado. Os tratamentos utilizados foram os herbicidas imazethapyr (75 g L-1 + imazapic (25 g L-1 nas doses de 0, 100, 150 e 200 g ha-1 do produto comercial Only®, acrescido de 0,5% do adjuvante Dash®. As doses foram aplicadas nos estádios V3-V4 do arroz CL. As variáveis estudadas foram: massa seca da parte aérea, peso de mil grãos e produtividade, nos experimentos em campo (EC1 e EC3, e altura de plantas, massa seca da parte aérea e massa seca de raiz, nos experimentos em casa de vegetação (CV1 e CV3. Os resultados demonstraram que o sistema CL de arroz irrigado pode restringir o cultivo em safras que intercalam cultivares tolerantes e não tolerantes ao herbicida Only, uma vez que a dose de 100 g ha¹, recomendada para controlar plantas daninhas nesse sistema de produção, deixa resíduo no solo em quantidade suficiente para interferir negativamente na safra seguinte, em relação ao crescimento e à produtividade do arroz convencional, cultivar IRGA 417.The aim of this work was to evaluate the residual soil activity of the herbicide Only (imazethapyr+imazapic to susceptible rice, cv. IRGA 417, after one or three years of Clearfield® (CL rice cultivation. Four experiments were conducted at the

  9. Oxidation of the tryptophan 32 residue of human superoxide dismutase 1 caused by its bicarbonate-dependent peroxidase activity triggers the non-amyloid aggregation of the enzyme.

    Science.gov (United States)

    Coelho, Fernando R; Iqbal, Asif; Linares, Edlaine; Silva, Daniel F; Lima, Filipe S; Cuccovia, Iolanda M; Augusto, Ohara

    2014-10-31

    The role of oxidative post-translational modifications of human superoxide dismutase 1 (hSOD1) in the amyotrophic lateral sclerosis (ALS) pathology is an attractive hypothesis to explore based on several lines of evidence. Among them, the remarkable stability of hSOD1(WT) and several of its ALS-associated mutants suggests that hSOD1 oxidation may precede its conversion to the unfolded and aggregated forms found in ALS patients. The bicarbonate-dependent peroxidase activity of hSOD1 causes oxidation of its own solvent-exposed Trp(32) residue. The resulting products are apparently different from those produced in the absence of bicarbonate and are most likely specific for simian SOD1s, which contain the Trp(32) residue. The aims of this work were to examine whether the bicarbonate-dependent peroxidase activity of hSOD1 (hSOD1(WT) and hSOD1(G93A) mutant) triggers aggregation of the enzyme and to comprehend the role of the Trp(32) residue in the process. The results showed that Trp(32) residues of both enzymes are oxidized to a similar extent to hSOD1-derived tryptophanyl radicals. These radicals decayed to hSOD1-N-formylkynurenine and hSOD1-kynurenine or to a hSOD1 covalent dimer cross-linked by a ditryptophan bond, causing hSOD1 unfolding, oligomerization, and non-amyloid aggregation. The latter process was inhibited by tempol, which recombines with the hSOD1-derived tryptophanyl radical, and did not occur in the absence of bicarbonate or with enzymes that lack the Trp(32) residue (bovine SOD1 and hSOD1(W32F) mutant). The results support a role for the oxidation products of the hSOD1-Trp(32) residue, particularly the covalent dimer, in triggering the non-amyloid aggregation of hSOD1. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Carbons prepared from Spartina alterniflora and its anaerobically digested residue by H{sub 3}PO{sub 4} activation: Characterization and adsorption of cadmium from aqueous solutions

    Energy Technology Data Exchange (ETDEWEB)

    Wang Zhengfang; Nie Er; Li Jihua; Zhao Yongjun; Luo Xingzhang [Environmental Science and Engineering Department, Fudan University, Shanghai 200433 (China); Zheng Zheng, E-mail: zzhenghj@fudan.edu.cn [Environmental Science and Engineering Department, Fudan University, Shanghai 200433 (China)

    2011-04-15

    Two series of activated carbons were prepared from Spartina alterniflora and from its anaerobically digested residue by H{sub 3}PO{sub 4} activation at various process conditions, and used as adsorbents for the removal of cadmium (II) in aqueous solutions. The surface areas and pore volumes of carbons were derived from adsorption isotherms (N{sub 2} at 77 K). The surface chemistry of carbons was investigated by infrared spectroscopy. Comparison study indicated that physicochemical properties of the activated carbons were strongly dependent not only on activation conditions but also on biopolymer contents of precursors. Several isotherm models were investigated and the adsorption isotherm data were best represented by the Langmuir isotherm model, with a maximum monolayer adsorption capacity of 47.85 mg/g at 25 deg. C. The results showed that the activated carbon produced from S. alterniflora could be employed as a promising adsorbent for removing cadmium (II) from aqueous solutions.

  11. Differences in the roles of conserved glutamic acid residues in the active site of human class 3 and class 2 aldehyde dehydrogenases.

    Science.gov (United States)

    Mann, C J; Weiner, H

    1999-10-01

    Although the three-dimensional structure of the dimeric class 3 rat aldehyde dehydrogenase has recently been published (Liu ZJ et al., 1997, Nature Struct Biol 4:317-326), few mechanistic studies have been conducted on this isoenzyme. We have characterized the enzymatic properties of recombinant class 3 human stomach aldehyde dehydrogenase, which is very similar in amino acid sequence to the class 3 rat aldehyde dehydrogenase. We have determined that the rate-limiting step for the human class 3 isozyme is hydride transfer rather than deacylation as observed for the human liver class 2 mitochondrial enzyme. No enhancement of NADH fluorescence was observed upon binding to the class 3 enzyme, while fluorescence enhancement of NADH has been previously observed upon binding to the class 2 isoenzyme. It was also observed that binding of the NAD cofactor inhibited the esterase activity of the class 3 enzyme while activating the esterase activity of the class 2 enzyme. Site-directed mutagenesis of two conserved glutamic acid residues (209 and 333) to glutamine residues indicated that, unlike in the class 2 enzyme, Glu333 served as the general base in the catalytic reaction and E209Q had only marginal effects on enzyme activity, thus confirming the proposed mechanism (Hempel J et al., 1999, Adv Exp Med Biol 436:53-59). Together, these data suggest that even though the subunit structures and active site residues of the isozymes are similar, the enzymes have very distinct properties besides their oligomeric state (dimer vs. tetramer) and substrate specificity.

  12. Residue Y70 of the Nitrilase Cyanide Dihydratase fromBacillus pumilusIs Critical for Formation and Activity of the Spiral Oligomer.

    Science.gov (United States)

    Park, Jason M; Ponder, Christian M; Sewell, B Trevor; Benedik, Michael J

    2016-12-28

    Nitrilases pose attractive alternatives to the chemical hydrolysis of nitrile compounds. The activity of bacterial nitrilases towards substrate is intimately tied to the formation of large spiral-shaped oligomers. In the nitrilase CynD (cyanide dihydratase) from Bacillus pumilus , mutations in a predicted oligomeric surface region altered its oligomerization and reduced its activity. One mutant, CynD Y70C, retained uniform oligomer formation however it was inactive, unlike all other inactive mutants throughout that region all of which significantly perturbed oligomer formation. It was hypothesized that Y70 is playing an additional role necessary for CynD activity beyond influencing oligomerization. Here, we performed saturation mutagenesis at residue 70 and demonstrated that only tyrosine or phenylalanine is permissible for CynD activity. Furthermore, we show that other residues at this position are not only inactive, but have altered or disrupted oligomer conformations. These results suggest that Y70's essential role in activity is independent of its role in the formation of the spiral oligomer.

  13. Designing with residual materials

    NARCIS (Netherlands)

    Walhout, W.; Wever, R.; Blom, E.; Addink-Dölle, L.; Tempelman, E.

    2013-01-01

    Many entrepreneurial businesses have attempted to create value based on the residual material streams of third parties. Based on ‘waste’ materials they designed products, around which they built their company. Such activities have the potential to yield sustainable products. Many of such companies

  14. H-bonding networks of the distal residues and water molecules in the active site of Thermobifida fusca hemoglobin.

    OpenAIRE

    Nicoletti , Francesco P; Droghetti , Enrica; Howes , Barry D; Bustamante , Juan P; Bonamore , Alessandra; Sciamanna , Natascia; Estrin , Darío A; Feis , Alessandro; Boffi , Alberto; Smulevich , Giulietta

    2013-01-01

    International audience; The ferric form of truncated hemoglobin II from Thermobifida fusca (Tf-trHb) and its triple mutant WG8F-YB10F-YCD1F at neutral and alkaline pH, and in the presence of CN(-) have been characterized by resonance Raman spectroscopy, electron paramagnetic resonance spectroscopy, and molecular dynamics simulations. Tf-trHb contains three polar residues in the distal site, namely TrpG8, TyrCD1 and TyrB10. Whereas TrpG8 can act as a potential hydrogen-bond donor, the tyrosine...

  15. Terapi Aktifitas Kelompok (Tak): Stimulasi Persepsi Modifikasi sebagai Alternatif Pengendalian Halusinasi Dengar pada Klien Skizofrenia

    OpenAIRE

    Ah. Yusuf, Ah; Fitryasari, Rizki; Nursalam, Nursalam; Iskandar, Iskandar

    2007-01-01

    The objective of this study was to examine the effect of modified Group Activity Therapy (GAT) perception stimulation on auditory halucination controlling in schizophrenia. Modification means using only 3 sessions in GAT. A quasi-experimental pre post test design was used in this study. Eighteen samples with auditory halucination at male third's class wards Menur Mental Hospital Surabaya were used in this study and divided into 2 groups. Observation was applied to measure patient's respons...

  16. Suitable conditions for xylanases activities from Bacillus sp. GA2(1 and Bacillus sp. GA1(6 and their properties for agricultural residues hydrolysis

    Directory of Open Access Journals (Sweden)

    Sudathip Chantorn

    2016-04-01

    Full Text Available Bacillus sp. GA2(1 and Bacillus sp. GA1(6 were isolated from soybean field in Khon Kaen province, Thailand. Crude enzymes from both isolates showed the activities of cellulase, xylanase, and mannanase at 37°C for 24 h. The highest xylanase activities of Bacillus sp. GA2(1 and Bacillus sp. GA1(6 were 1.58±0.25 and 0.82±0.16 U/ml, respectively. The relative xylanase activities from both strains were more than 60% at pH 5.0 to 8.0. The optimum temperature of xylanases was 50°C in both strains. The residual xylanase activities from both strains were more than 70% at 60°C for 60 min. Five agricultural wastes (AWs, namely coffee residue, soybean meal, potato peel, sugarcane bagasse, and corn cobs, were used as substrates for hydrolysis properties. The highest reducing sugar content of 101±1.32 µg/ml was obtained from soybean meal hydrolysate produced by Bacillus sp. GA2(1 xylanase.

  17. Analysis of active site residues of botulinum neurotoxin E by mutational, functional, and structural studies: Glu335Gln is an apoenzyme.

    Science.gov (United States)

    Agarwal, Rakhi; Binz, Thomas; Swaminathan, Subramanyam

    2005-06-14

    Clostridial neurotoxins comprising the seven serotypes of botulinum neurotoxins and tetanus neurotoxin are the most potent toxins known to humans. Their potency coupled with their specificity and selectivity underscores the importance in understanding their mechanism of action in order to develop a strategy for designing counter measures against them. To develop an effective vaccine against the toxin, it is imperative to achieve an inactive form of the protein which preserves the overall conformation and immunogenicity. Inactive mutants can be achieved either by targeting active site residues or by modifying the surface charges farther away from the active site. The latter affects the long-range forces such as electrostatic potentials in a subtle way without disturbing the structural integrity of the toxin causing some drastic changes in the activity/environment. Here we report structural and biochemical analysis on several mutations on Clostridium botulinum neurotoxin type E light chain with at least two producing dramatic effects: Glu335Gln causes the toxin to transform into a persistent apoenzyme devoid of zinc, and Tyr350Ala has no hydrolytic activity. The structural analysis of several mutants has led to a better understanding of the catalytic mechanism of this family of proteins. The residues forming the S1' subsite have been identified by comparing this structure with a thermolysin-inhibitor complex structure.

  18. Autophosphorylation of Specific Threonine and Tyrosine Residues in Arabidopsis CERK1 is Essential for the Activation of Chitin-Induced Immune Signaling.

    Science.gov (United States)

    Suzuki, Maruya; Shibuya, Masatoshi; Shimada, Hikaru; Motoyama, Noriko; Nakashima, Masato; Takahashi, Shohei; Suto, Kenkichi; Yoshida, Issei; Matsui, Saki; Tsujimoto, Natsumi; Ohnishi, Mihoko; Ishibashi, Yuko; Fujimoto, Zui; Desaki, Yoshitake; Kaku, Hanae; Kito, Keiji; Shibuya, Naoto

    2016-11-01

    Pattern recognition receptors on the plant cell surface mediate the recognition of microbe/damage-associated molecular patterns (MAMPs/DAMPs) and activate downstream immune signaling. Autophosphorylation of signaling receptor-like kinases is a critical event for the activation of downstream responses but the function of each phosphorylation site in the regulation of immune signaling is not well understood. In this study, 41 Ser/Thr/Tyr and 15 Ser/Thr residues were identified as in vitro and in vivo autophosphorylation sites of Arabidopsis CERK1, which is essential for chitin signaling. Comprehensive analysis of transgenic plants expressing mutated CERK1 genes for each phosphorylation site in the cerk1-2 background indicated that the phosphorylation of T479 in the activation segment and Y428 located upstream of the catalytic loop is important for the activation of chitin-triggered defense responses. Contribution of the phosphorylation of T573 to the chitin responses was also suggested. In vitro evaluation of kinase activities of mutated kinase domains indicated that the phosphorylation of T479 and T573 is directly involved in the regulation of kinase activity of CERK1 but the phosphorylation of Y428 regulates chitin signaling independently of the regulation of kinase activity. These results indicated that the phosphorylation of specific residues in the kinase domain contributes to the regulation of downstream signaling either through the regulation of kinase activity or the different mechanisms, e.g. regulation of protein-protein interactions. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  19. Effect of a non-woven fabric covering on the residual activity of pendimethalin in lettuce and soil.

    Science.gov (United States)

    Jursík, Miroslav; Kováčová, Jana; Kočárek, Martin; Hamouzová, Kateřina; Soukup, Josef

    2017-05-01

    Lettuce (Lactuca sativa L.) is a crop that is very sensitive to herbicide contamination owing to its short growing season. The use of long-residual herbicides and non-woven fabric coverings could therefore influence pendimethalin concentrations in soil and lettuce. The pendimethalin half-life in soil ranged between 18 and 85 days and was mainly affected by season (i.e. weather), and especially by soil moisture. Pendimethalin degradation in soil was slowest under dry conditions. A longer pendimethalin half-life was observed under the non-woven fabric treatment, but the effect of varying application rate was not significant. Pendimethalin residue concentrations in lettuce heads were significantly influenced by pendimethalin application rate and by non-woven fabric cover, especially at the lettuce's early growth stages. The highest pendimethalin concentration at final harvest was determined in lettuce grown on uncovered plots treated with pendimethalin at an application rate of 1200 g ha -1 (7-38 µg kg -1 ). Depending on growing season duration and weather conditions, pendimethalin concentrations in lettuce grown under non-woven fabric ranged from 0 to 21 µg kg -1 . Use of transparent non-woven fabric cover with lettuce can help to reduce application rates of soil herbicides and diminish the risk of herbicide contamination in the harvested vegetables. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  20. Terapi Aktifitas Kelompok (TAK): Stimulasi Persepsi Modifikasi sebagai Alternatif Pengendalian Halusinasi Dengar pada Klien Skizofrenia

    OpenAIRE

    Ah. Yusuf, Ah.; Fitryasari, Rizki; Nursalam, Nursalam; Iskandar, Iskandar

    2017-01-01

    The objective of this study was to examine the effect of modified Group Activity Therapy (GAT) perception stimulation on auditory halucination controlling in schizophrenia. Modification means using only 3 sessions in GAT. A quasi-experimental pre post test design was used in this study.   Eighteen  samples with auditory halucination at male third’s class wards Menur Mental Hospital Surabaya were used in this study and divided into 2 groups. Observation was applied to measure patient’s respons...

  1. RESIDUAL RISK ASSESSMENTS - RESIDUAL RISK ...

    Science.gov (United States)

    This source category previously subjected to a technology-based standard will be examined to determine if health or ecological risks are significant enough to warrant further regulation for Coke Ovens. These assesments utilize existing models and data bases to examine the multi-media and multi-pollutant impacts of air toxics emissions on human health and the environment. Details on the assessment process and methodologies can be found in EPA's Residual Risk Report to Congress issued in March of 1999 (see web site). To assess the health risks imposed by air toxics emissions from Coke Ovens to determine if control technology standards previously established are adequately protecting public health.

  2. Characterization of human paraoxonase 1 variants suggest that His residues at 115 and 134 positions are not always needed for the lactonase/arylesterase activities of the enzyme

    Science.gov (United States)

    Bajaj, Priyanka; Tripathy, Rajan K; Aggarwal, Geetika; Pande, Abhay H

    2013-01-01

    Human paraoxonase 1 (h-PON1) hydrolyzes variety of substrates and the hydrolytic activities of enzyme can be broadly grouped into three categories; arylesterase, phosphotriesterase, and lactonase. Current models of the catalytic mechanism of h-PON1 suggest that catalytic residues H115 and H134 mediate the lactonase and arylesterase activities of the enzyme. H-PON1 is a strong candidate for the development of catalytic bioscavenger for organophosphate poisoning in humans. Recently, Gupta et al. (Nat. Chem. Biol. 2011. 7, 120) identified amino acid substitutions that significantly increased the activity of chimeric-PON1 variant (4E9) against some organophosphate nerve agents. In this study we have examined the effect of these (L69G/S111T/H115W/H134R/R192K/F222S/T332S) and other substitutions (H115W/H134R and H115W/H134R/R192K) on the hydrolytic activities of recombinant h-PON1 (rh-PON1) variants. Our results show that the substitutions resulted in a significant increase in the organophosphatase activity of all the three variants of rh-PON1 enzyme while had a variable effect on the lactonase/arylesterase activities. The results suggest that H residues at positions 115 and 134 are not always needed for the lactonase/arylesterase activities of h-PON1 and force a reconsideration of the current model(s) of the catalytic mechanism of h-PON1. PMID:24123308

  3. TERAPI AKTIFITAS KELOMPOK (TAK: STIMULASI PERSEPSI MODIFIKASI SEBAGAI ALTERNATIF PENGENDALIAN HALUSINASI DENGAR PADA KLIEN SKIZOFRENIA

    Directory of Open Access Journals (Sweden)

    Ah. Ah. Yusuf

    2017-07-01

    Full Text Available The objective of this study was to examine the effect of modified Group Activity Therapy (GAT perception stimulation on auditory halucination controlling in schizophrenia. Modification means using only 3 sessions in GAT. A quasi-experimental pre post test design was used in this study.   Eighteen  samples with auditory halucination at male third’s class wards Menur Mental Hospital Surabaya were used in this study and divided into 2 groups. Observation was applied to measure patient’s responses and data were analyzed by Wilcoxon Signed Rank test and Mann Whitney test with significance level of  α<0.05. Conclusion:  modified perception stimulation of GAT is able to increase patient’s ability for making differences between reality and non-reality and motivated patient to choose and use the best way  for controlling halucination.

  4. Scintigraphic criteria for administration of therapeutic activities of 131I for ablation of residual thyroid tissue in patients with differentiated thyroid cancer

    International Nuclear Information System (INIS)

    Dzhuzha, D.A.

    2003-01-01

    With the aim to determinate the optical scintigraphic criteria of radioablation of the small parts of residual thyroid tissue (RTT) after a thyroidectomy the results of postoperative monitoring of 29 women aged 23-60 with differentiated thyroid cancer, who did not get the radioiodine therapy, were analyzed. At the single postoperative scintigraphy with 131 I the optimal scintigraphic criteria for radioablation are the relative level of 131 I accumulation in RTT and the relative specific volume activity in RTT. At the values of these indices below 0.21% and 0.16%/sm 3 , respectively, only suppressive thyroid hormone therapy and standard monitoring are recommended

  5. Pathogen response-like recruitment and activation of neutrophils by sterile immunogenic dying cells drives neutrophil-mediated residual cell killing.

    Science.gov (United States)

    Garg, Abhishek D; Vandenberk, Lien; Fang, Shentong; Fasche, Tekele; Van Eygen, Sofie; Maes, Jan; Van Woensel, Matthias; Koks, Carolien; Vanthillo, Niels; Graf, Norbert; de Witte, Peter; Van Gool, Stefaan; Salven, Petri; Agostinis, Patrizia

    2017-05-01

    Innate immune sensing of dying cells is modulated by several signals. Inflammatory chemokines-guided early recruitment, and pathogen-associated molecular patterns-triggered activation, of major anti-pathogenic innate immune cells like neutrophils distinguishes pathogen-infected stressed/dying cells from sterile dying cells. However, whether certain sterile dying cells stimulate innate immunity by partially mimicking pathogen response-like recruitment/activation of neutrophils remains poorly understood. We reveal that sterile immunogenic dying cancer cells trigger (a cell autonomous) pathogen response-like chemokine (PARC) signature, hallmarked by co-release of CXCL1, CCL2 and CXCL10 (similar to cells infected with bacteria or viruses). This PARC signature recruits preferentially neutrophils as first innate immune responders in vivo (in a cross-species, evolutionarily conserved manner; in mice and zebrafish). Furthermore, key danger signals emanating from these dying cells, that is, surface calreticulin, ATP and nucleic acids stimulate phagocytosis, purinergic receptors and toll-like receptors (TLR) i.e. TLR7/8/9-MyD88 signaling on neutrophil level, respectively. Engagement of purinergic receptors and TLR7/8/9-MyD88 signaling evokes neutrophil activation, which culminates into H 2 O 2 and NO-driven respiratory burst-mediated killing of viable residual cancer cells. Thus sterile immunogenic dying cells perform 'altered-self mimicry' in certain contexts to exploit neutrophils for phagocytic targeting of dead/dying cancer cells and cytotoxic targeting of residual cancer cells.

  6. Cellulase with high β-glucosidase activity by Penicillium oxalicum under solid state fermentation and its use in hydrolysis of cassava residue.

    Science.gov (United States)

    Su, Lin-Hui; Zhao, Shuai; Jiang, Sui-Xin; Liao, Xu-Zhong; Duan, Cheng-Jie; Feng, Jia-Xun

    2017-02-01

    In this study, we investigated cellulase production by Penicillium oxalicum EU2106 under solid-state fermentation (SSF) and its hydrolysis efficiency toward NaOH-H 2 O 2 -pretreated cassava residue (NHCR) produced after bioethanol fermentation. Optimization of SSF cultivation conditions for P. oxalicum EU2106 using a Box-behnken design-based response-surface methodology resulted in maximal cellulase activity of 34.0 ± 2.8 filter-paper units/g dry substrate, exhibiting a ~ twofold increase relative to activities obtained under non-optimized conditions. Furthermore, SSF-derived cellulase converted 94.3 ± 1.5% of NHCR cellulose into glucose within 96 h. Interestingly, P. oxalicum EU2106 produced higher β-glucosidase activity under SSF conditions than that under submerged-state fermentation conditions, resulting in the elimination of cellobiose inhibition during the early stages of NHCR cellulose hydrolysis. Overall, this work provided an alternative for a potential cellulase source and a preferred option for cassava residue biotechnological application.

  7. Tank characterization report for single-shell tak 241-C-112. Revision 1

    Energy Technology Data Exchange (ETDEWEB)

    Simpson, B.C.

    1997-06-11

    One major function of the Tank Waste Remediation System (IWRS) is to characterize wastes in support of waste management and disposal activities at the Hanford Site. Analytical data from sampling and analysis and other available information about a tank are compiled and maintained in a tank characterization report (CR). This report and its appendixes serve as the CR for single-shell tank 24 1 -C- 1 12. The objectives of this report are: 1) to use characterization data in response to technical issues associated with tank 24 1 -C- 1 12 waste, and 2) to provide a standard characterization of this waste in terms of a best-basis inventory estimate. Section 2.0 summarizes the response to technical issues, Section 3.0 shows the best-basis inventory estimate, and Section 4.0 makes recommendations regarding safety status and additional sampling needs. The appendixes contain supporting data and information. This report supports the requirements of the Hanford Federal Facility Agreement and Consent Order, Milestone M-44-05 (Ecology et al. 1996).

  8. Residual nilpotence and residual solubility of groups

    International Nuclear Information System (INIS)

    Mikhailov, R V

    2005-01-01

    The properties of the residual nilpotence and the residual solubility of groups are studied. The main objects under investigation are the class of residually nilpotent groups such that each central extension of these groups is also residually nilpotent and the class of residually soluble groups such that each Abelian extension of these groups is residually soluble. Various examples of groups not belonging to these classes are constructed by homological methods and methods of the theory of modules over group rings. Several applications of the theory under consideration are presented and problems concerning the residual nilpotence of one-relator groups are considered.

  9. Transcatheter arterial embolization combined with radiofrequency ablation activates CD8+ T-cell infiltration surrounding residual tumors in the rabbit VX2 liver tumors

    Directory of Open Access Journals (Sweden)

    Duan XH

    2016-05-01

    Full Text Available Xu-Hua Duan,1,2 Teng-Fei Li,2 Guo-Feng Zhou,1,* Xin-Wei Han,2,* Chuan-Sheng Zheng,1 Peng-fei Chen,2 Gan-Sheng Feng11Department of Interventional Radiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 2Department of Interventional Radiology, The First Affiliated Hospital, Zhengzhou University, Henan Province, Zhengzhou, People’s Republic of China*These authors contributed equally to this work Purpose: To evaluate the effect of transcatheter arterial embolization (TAE combined with radiofrequency ablation (RFA treatment (TAE + RFA on the expression of heat shock protein 70 (HSP70 in residual tumors and explore the relationship between the HSP70 and CD8+ T-cell infiltrate surrounding residual tumors in the rabbit VX2 liver tumor model.Materials and methods: Animals with VX2 liver tumors were randomized into four groups (control, TAE, RFA, and TAE + RFA with 15 rabbits in each group. Five rabbits in each group were sacrificed on days 1, 3, and 7 after treatment. HSP70 expression and infiltration of CD8+ T-cells in the liver and residual tumors surrounding the necrosis zone were detected by immunohistochemistry staining. The maximal diameters of tumor necrosis, numbers of metastases, and tumor growth rate were compared on day 7 after treatment.Results: TAE + RFA achieved larger maximal diameter of tumor necrosis, lower tumor growth rate, and fewer metastatic lesions, compared with other treatments on day 7. The number of CD8+ T-cells in the TAE + RFA group was significantly higher than in other groups on days 1, 3, and 7. There was a positive correlation between HSP70 expression level and infiltration of CD8+ T-cells surrounding the residual tumor on day 1 (r=0.9782, P=0.012, day 3 (r=0.93, P=0.021, and day 7 (r=0.8934, P=0.034.Conclusion: In the rabbit VX2 liver tumor model, TAE + RFA activated the highest number of CD8+ T-cells surrounding residual tumors. TAE + RFA appears to be a beneficial

  10. Preparation of a Nanoemulsion with Carapa guianensis Aublet (Meliaceae Oil by a Low-Energy/Solvent-Free Method and Evaluation of Its Preliminary Residual Larvicidal Activity

    Directory of Open Access Journals (Sweden)

    Flávia L. M. Jesus

    2017-01-01

    Full Text Available Andiroba (Carapa guianensis seeds are the source of an oil with a wide range of biological activities and ethnopharmacological uses. However, few studies have devoted attention to innovative formulations, including nanoemulsions. The present study aimed to obtain a colloidal system with the andiroba oil using a low-energy and organic-solvent-free method. Moreover, the preliminary residual larvicidal activity of the nanoemulsion against Aedes aegypti was evaluated. Oleic and palmitic acids were the major fatty acids, in addition to the phytosterol β-sitosterol and limonoids (tetranortriterpenoids. The required hydrophile-lipophile was around 11.0 and the optimal nanoemulsion was obtained using polysorbate 85. The particle size distribution suggested the presence of small droplets (mean diameter around 150 nm and low polydispersity index (around 0.150. The effect of temperature on particle size distribution revealed that no major droplet size increase occurred. The preliminary residual larvicidal assay suggested that the mortality increased as a function of time. The present study allowed achievement of a potential bioactive oil in water nanoemulsion that may be a promising controlled release system. Moreover, the ecofriendly approach involved in the preparation associated with the great bioactive potential of C. guianensis makes this nanoemulsion very promising for valorization of this Amazon raw material.

  11. The Strictly Conserved Arg-321 Residue in the Active Site of Escherichia coli Topoisomerase I Plays a Critical Role in DNA Rejoining*

    Science.gov (United States)

    Narula, Gagandeep; Annamalai, Thirunavukkarasu; Aedo, Sandra; Cheng, Bokun; Sorokin, Elena; Wong, Agnes; Tse-Dinh, Yuk-Ching

    2011-01-01

    The strictly conserved arginine residue proximal to the active site tyrosine of type IA topoisomerases is required for the relaxation of supercoiled DNA and was hypothesized to be required for positioning of the scissile phosphate for DNA cleavage to take place. Mutants of recombinant Yersinia pestis topoisomerase I with hydrophobic substitutions at this position were found in genetic screening to exhibit a dominant lethal phenotype, resulting in drastic loss in Escherichia coli viability when overexpressed. In depth biochemical analysis of E. coli topoisomerase I with the corresponding Arg-321 mutation showed that DNA cleavage can still take place in the absence of this arginine function if Mg2+ is present to enhance the interaction of the enzyme with the scissile phosphate. However, DNA rejoining is inhibited in the absence of this conserved arginine, resulting in accumulation of the cleaved covalent intermediate and loss of relaxation activity. These new experimental results demonstrate that catalysis of DNA rejoining by type IA topoisomerases has a more stringent requirement than DNA cleavage. In addition to the divalent metal ions, the side chain of this arginine residue is required for the precise positioning of the phosphotyrosine linkage for nucleophilic attack by the 3′-OH end to result in DNA rejoining. Small molecules that can interfere or distort the enzyme-DNA interactions required for DNA rejoining by bacterial type IA topoisomerases could be developed into novel antibacterial drugs. PMID:21478161

  12. Potential active-site residues in polyneuridine aldehyde esterase, a central enzyme of indole alkaloid biosynthesis, by modelling and site-directed mutagenesis.

    Science.gov (United States)

    Mattern-Dogru, Emine; Ma, Xueyan; Hartmann, Joachim; Decker, Heinz; Stöckigt, Joachim

    2002-06-01

    In the biosynthesis of the antiarrhythmic alkaloid ajmaline, polyneuridine aldehyde esterase (PNAE) catalyses a central reaction by transforming polyneuridine aldehyde into epi-vellosimine, which is the immediate precursor for the synthesis of the ajmalane skeleton. The PNAE cDNA was previously heterologously expressed in E. coli. Sequence alignments indicated that PNAE has a 43% identity to a hydroxynitrile lyase from Hevea brasiliensis, which is a member of the alpha/beta hydrolase superfamily. The catalytic triad, which is typical for this family, is conserved. By site-directed mutagenesis, the members of the catalytic triad were identified. For further detection of the active residues, a model of PNAE was constructed based on the X-ray crystallographic structure of hydroxynitrile lyase. The potential active site residues were selected on this model, and were mutated in order to better understand the relationship of PNAE with the alpha/beta hydrolases, and as well its mechanism of action. The results showed that PNAE is a novel member of the alpha/beta hydrolase enzyme superfamily.

  13. Andreas Fülberth, „Riga. Kleine Geschichte der Stadt”. Ciekawy przewodnik nie tylko po Rydze, ale także po historii Łotwy

    Directory of Open Access Journals (Sweden)

    Tomasz Kamil Otocki

    2014-12-01

    At the beginning of 2014, the book „Riga. Kleine Geschichte der Stadt” (Riga. A short history of the town was issued. The author is Andreas Fülberth, a young historian from Germany, who is a lecturer of the history of Eastern Europe in the University of Kiel. He has already published several works about the Baltic states (in German: “Baltikum”, the most important of them being „Tallinn – Riga – Kaunas. Ihr Ausbau zu modernen Hauptstädten 1920-1940". Köln u. a. 2005 (Das Baltikum in Geschichte und Gegenwart, Bd. 2, which is dedicated to the plans of architectural rebuilding of the Baltic capitals (Kaunas – Riga – Tallinn during the time of the first independence (1918-1940. The history of Riga by Andreas Fülberth begins – very traditionally – with the establishing of the town by Bishop Albert of Riga in 1201. Actually we can learn not only about the history of the town. The book by Andreas Fülberth provides a quite long trip through the history of Livonia (now a part of Latvia. For Polish readers very important and interesting piece of  Riga’s history could be so called “Polish times” (to be more precise: “Polish-Lithuanian” in Livonia – which used to be seen quite critical by Latvian historians before the war. We can learn also about Ignacy Mościcki  who studied in Riga, the treaty of Riga from March 1921,  as well  as the Polish academic fraternities in Livonia (Arcadia and Welecja. Maybe the most important part of the book begins in 1918 – when Latvia gained independence for the first time in her history. We can learn not only about Kārlis Ulmanis and the Soviet-Latvian government of Pēteris Stučka, but also about  the activities of Andrievs Niedra, a pro-German prime minister of Latvia. Andreas Fülberth, as a passionate lover  of  architecture, provides an interesting piece of information about the architectural rebuilding of Riga during the time of Ulmanis – we can learn the history of the Freedom Monument

  14. Access to free or low-cost tuberculosis treatment for migrants and refugees along the Thailand-Myanmar border in Tak province, Thailand.

    Science.gov (United States)

    Tschirhart, Naomi; Nosten, Francois; Foster, Angel M

    2016-07-07

    In Tak province, Thailand migrants and refugees from Myanmar navigate a pluralistic healthcare system to seek Tuberculosis (TB) care from a variety of government and non-governmental providers. This multi-methods qualitative study examined access to TB, TB/HIV and multidrug-resistant tuberculosis (MDR-TB) treatment with an emphasis on barriers to care and enabling factors. In the summer and fall of 2014, we conducted 12 key informant interviews with public health officials and TB treatment providers. We also conducted 11 focus group discussions with migrants and refugees who were receiving TB, TB/HIV and MDR-TB treatment in Tak province as well as non-TB patients. We analyzed these data through thematic analysis using both predetermined and emergent codes. As a second step in the qualitative analysis, we explored the barriers and enabling factors separately for migrants and refugees. We found that refugees face fewer barriers to accessing TB treatment than migrants. For both migrants and refugees, legal status plays an important intermediary role in influencing the population's ability to access care and eligibility for treatment. Our results suggest that there is a large geographical catchment area for migrants who seek TB treatment in Tak province that extends beyond provincial boundaries. Migrant participants described their ability to seek care as linked to the financial and non-financial resources required to travel and undergo treatment. Patients identified language of health services, availability of free or low cost services, and psychosocial support as important health system characteristics that affect accessibility. Access to TB treatment for migrants and refugees occurs at the interface of health system accessibility, population ability and legal status. In Tak province, migrant patients draw upon their social networks and financial resources to navigate a pathway to treatment. We revised a conceptual framework for access to healthcare to incorporate

  15. Subcritical water extraction and antioxidant activity evaluation with on-line HPLC-ABTS(·+) assay of phenolic compounds from marigold (Tagetes erecta L.) flower residues.

    Science.gov (United States)

    Xu, Honggao; Wang, Weiyou; Jiang, Junping; Yuan, Fang; Gao, Yanxiang

    2015-06-01

    Subcritical water extraction (SWE) of phenolics was investigated from marigold (Tagetes erecta L.) flower residues. The total phenolics content (TPC), total flavonoids content (TFC) and antioxidant capacities of extracts were determined, furthermore, antioxidant activities of individual compounds were evaluated with on-line HPLC-ABTS(•+) system. The optimum SWE time was 45 min, solid-to-liquid ratio was 1:50, and the highest TPC and TFC were obtained at 220 °C respectively. The effect of SWE temperature on TPC and TFC was significant (p line HPLC-ABTS(•+) profiles revealed that quercetagetin from SWE at 200 °C had nearly twofold radical scavenging activities than that by leaching extraction.

  16. Evaluation of residue-residue contact predictions in CASP9

    KAUST Repository

    Monastyrskyy, Bohdan

    2011-01-01

    This work presents the results of the assessment of the intramolecular residue-residue contact predictions submitted to CASP9. The methodology for the assessment does not differ from that used in previous CASPs, with two basic evaluation measures being the precision in recognizing contacts and the difference between the distribution of distances in the subset of predicted contact pairs versus all pairs of residues in the structure. The emphasis is placed on the prediction of long-range contacts (i.e., contacts between residues separated by at least 24 residues along sequence) in target proteins that cannot be easily modeled by homology. Although there is considerable activity in the field, the current analysis reports no discernable progress since CASP8.

  17. Residual limb fluid volume change and volume accommodation: Relationships to activity and self-report outcomes in people with trans-tibial amputation.

    Science.gov (United States)

    Sanders, Joan E; Youngblood, Robert T; Hafner, Brian J; Ciol, Marcia A; Allyn, Katheryn J; Gardner, David; Cagle, John C; Redd, Christian B; Dietrich, Colin R

    2018-02-01

    Fluctuations in limb volume degrade prosthesis fit and require users to accommodate changes using management strategies, such as donning and doffing prosthetic socks. To examine how activities and self-report outcomes relate to daily changes in residual limb fluid volume and volume accommodation. Standardized, two-part laboratory protocol with an interim observational period. Participants were classified as "accommodators" or "non-accommodators," based on self-report prosthetic sock use. Participants' residual limb fluid volume change was measured using a custom bioimpedance analyzer and a standardized in-laboratory activity protocol. Self-report health outcomes were assessed with the Socket Comfort Score and Prosthesis Evaluation Questionnaire. Activity was monitored while participants left the laboratory for at least 3 h. They then returned to repeat the bioimpedance test protocol. Twenty-nine people were enrolled. Morning-to-afternoon percent limb fluid volume change per hour was not strongly correlated to percent time weight-bearing or to self-report outcomes. As a group, non-accommodators ( n = 15) spent more time with their prosthesis doffed and reported better outcomes than accommodators. Factors other than time weight-bearing may contribute to morning-to-afternoon limb fluid volume changes and reported satisfaction with the prosthesis among trans-tibial prosthesis users. Temporary doffing may be a more effective and satisfying accommodation method than sock addition. Clinical relevance Practitioners should be mindful that daily limb fluid volume change and prosthesis satisfaction are not dictated exclusively by activity. Temporarily doffing the prosthesis may slow daily limb fluid volume loss and should be investigated as an alternative strategy to sock addition.

  18. A single amino acid residue, Ala 105, confers 16alpha-hydroxylase activity to human cytochrome P450 17alpha-hydroxylase/17,20 lyase.

    Science.gov (United States)

    Swart, Amanda C; Storbeck, Karl-Heinz; Swart, Pieter

    2010-04-01

    In adrenal steroidogenesis, CYP17 catalyses the 17alpha-hydroxylation of pregnenolone and progesterone and the subsequent 17,20-lyase reaction, yielding adrenal androgens. The enzyme exhibits distinctly different selectivities towards these substrates in various species. CYP17 has also been shown to exhibit 16alpha-hydroxylase activity towards progesterone in some species, with only human and chimp CYP17 catalysing the biosynthesis of substantial amounts of 16-OHprogesterone. The 16alpha-hydroxylase activity was investigated by introducing an Ala105Leu substitution into human CYP17. The converse mutation, Leu105Ala was introduced into the baboon, goat and pig enzymes. Wt human CYP17 converted approximately 30% progesterone to 16-OHprogesterone while the Ala105Leu mutant converted negligible amounts to 16-OHprogesterone ( approximately 9%), comparable to wt CYP17 of the other three species when expressed in COS-1 cells. The ratio of 17-hydroxylated products to 16-OHprogesterone of human CYP17 was 2.7 and that of the mutant human construct 10.5. Similar ratios were observed for human and goat CYP17 with the corresponding Ala or Leu residues. Although the Leu105Ala mutation of both baboon and pig CYP17 exhibited the same trend regarding the ratios, the rate of progesterone conversion was reduced. Coexpression with cytochrome b(5) significantly decreased the ratio of 17-hydroxylated products to 16-OHprogesterone in the Leu105 constructs, while effects were negligible with Ala at this position. Homology models show that Ala105 faces towards the active pocket in the predicted B'-C domain of CYP17. The smaller residue allows more flexibility of movement in the active pocket than Leu, presenting both the C16 and C17 of progesterone to the iron-oxy complex. 2010 Elsevier Ltd. All rights reserved.

  19. Meget mere gaming tak

    DEFF Research Database (Denmark)

    Gamborg, Maria Louise; Lieberoth, Andreas; Mehlsen, Mimi

    2018-01-01

    Videnskaben og medierne har skabt en fortælling om computerspillende unge, der mister sociale kompetencer eller bliver afhængige af at game. Men det er en stærk unuanceret fortælling. Tværtimod kan computerspil være adgangen til ægte, følelsesmæssigt udviklende, sociale fællesskaber, hvor gameres...... sociale kapital øges ved at spille sammen online. Det skal vi lade os inspirere af - fx når det gælder ældre...

  20. Effects of Vitamin D Receptor Activation and Dietary Sodium Restriction on Residual Albuminuria in CKD: The ViRTUE-CKD Trial.

    Science.gov (United States)

    Keyzer, Charlotte A; van Breda, G Fenna; Vervloet, Marc G; de Jong, Maarten A; Laverman, Gozewijn D; Hemmelder, Marc H; Janssen, Wilbert M T; Lambers Heerspink, Hiddo J; Kwakernaak, Arjan J; Bakker, Stephan J L; Navis, Gerjan; de Borst, Martin H

    2017-04-01

    Reduction of residual albuminuria during single-agent renin-angiotensin-aldosterone blockade is accompanied by improved cardiorenal outcomes in CKD. We studied the individual and combined effects of the vitamin D receptor activator paricalcitol (PARI) and dietary sodium restriction on residual albuminuria in CKD. In a multicenter, randomized, placebo (PLAC)-controlled, crossover trial, 45 patients with nondiabetic CKD stages 1-3 and albuminuria >300 mg/24 h despite ramipril at 10 mg/d and BPalbuminuria (geometric mean) was 1060 (95% confidence interval, 778 to 1443) mg/24 h during RS + PLAC and 990 (95% confidence interval, 755 to 1299) mg/24 h during RS + PARI ( P =0.20 versus RS + PLAC). LS + PLAC reduced albuminuria to 717 (95% confidence interval, 512 to 1005) mg/24 h ( P albuminuria to 683 (95% confidence interval, 502 to 929) mg/24 h ( P albuminuria reduction ( P =0.04 LS + PARI versus LS + PLAC). Dietary adherence was good as reflected by urinary excretion of 174±64 mmol Na + per day in the combined RS groups and 108±61 mmol Na + per day in the LS groups ( P albuminuria during fixed dose angiotensin-converting enzyme inhibition. The additional effect of PARI was small and nonsignificant. Copyright © 2017 by the American Society of Nephrology.

  1. Activated sludge mass reduction and biodegradability of the endogenous residues by digestion under different aerobic to anaerobic conditions: Comparison and modeling.

    Science.gov (United States)

    Martínez-García, C G; Fall, C; Olguín, M T

    2016-03-01

    This study was performed to identify suitable conditions for the in-situ reduction of excess sludge production by intercalated digesters in recycle-activated sludge (RAS) flow. The objective was to compare and model biological sludge mass reduction and the biodegradation of endogenous residues (XP) by digestion under hypoxic, aerobic, anaerobic, and five intermittent-aeration conditions. A mathematical model based on the heterotrophic endogenous decay constant (bH) and including the biodegradation of XP was used to fit the long-term data from the digesters to identify and estimate the parameters. Both the bH constant (0.02-0.05 d(-1)) and the endogenous residue biodegradation constant (bP, 0.001-0.004 d(-1)) were determined across the different mediums. The digesters with intermittent aeration cycles of 12 h-12 h and 5 min-3 h (ON/OFF) were the fastest, compared to the aerobic reactor. The study provides a basis for rating RAS-digester volumes to avoid the accumulation of XP in aeration tanks. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. A Tyrosine Residue Along with a Glutamic Acid of the Omega-Like Loop Governs the Beta-Lactamase Activity of MSMEG_4455 in Mycobacterium smegmatis.

    Science.gov (United States)

    Bansal, Ankita; Kar, Debasish; Pandey, Satya Deo; Matcha, Ashok; Kumar, N Ganesh; Nathan, Soshina; Ghosh, Anindya S

    2017-06-01

    Mycobacterial beta-lactamases are involved in exerting beta-lactam resistance, though many of these proteins remain uncharacterized. Here, we have characterized MSMEG_4455 of Mycobacterium smegmatis as a beta-lactamase using molecular, biochemical and mutational techniques. To elucidate its nature in vivo and in vitro, and to predict its structure-function relationship in silico analysis is done. The MSMEG_4455 is cloned and expressed ectopically in a beta-lactamase deficient Escherichia coli mutant to establish the in vivo beta-lactamase like nature via minimum inhibitory concentration (MIC) determination. Likewise the in vivo results, purified soluble form of MSMEG_4455 showed beta-lactam hydrolysis pattern similar to group 2a penicillinase. In silico analyses of MSMEG_4455 reveal glutamic acid (E)193 and tyrosine (Y)194 of omega-like loop might have importance in strengthening hydrogen bond network around the active-site, though involvement of tyrosine is rare for beta-lactamase activity. Accordingly, these residues are mutated to alanine (A) and phenylalanine (F), respectively. The mutated proteins have partially lost their ability to exert beta-lactamase activity both in vivo and in vitro. The Y194F mutation had more prominent effect on the enzymatic activity. Therefore, we infer that Y194 is the key for beta-lactamase activity of MSMEG_4455.

  3. Understanding the role of residues around the active site tunnel towards generating a glucose-tolerant β-glucosidase from Agrobacterium tumefaciens 5A.

    Science.gov (United States)

    Goswami, Shubhasish; Das, Shibashis; Datta, Supratim

    2017-07-01

    Most β-glucosidases are subjected to inhibition by the final hydrolysis product glucose resulting in the accumulation of cellobiose and oligosaccharides. This accumulated cellobiose and oligosaccharides further inhibit the activities of endoglucanase and cellobiohydrolases, resulting in the inhibition of cellulose degradation and a more expensive biofuel. To elucidate the mechanism(s) of glucose tolerance, we designed and characterised six mutations of a moderately glucose-tolerant β-glucosidase (H0HC94) from the mesophilic bacterium Agrobacterium tumefaciens 5A. The hydrophobicity and steric were varied across non-conserved residues in specific regions of the active site tunnel. In contrast to the uncompetitive inhibition of WT enzyme by glucose, C174V and H229S are competitively inhibited pointing towards a possible glucose-binding site in the protein at these positions. Increasing hydrophobicity at the +1 subsite and increasing hydrophobicity and steric at +2 subsites seemed to be critical for glucose tolerance for this BG. Additionally, in L178E, specific activity was 1.8 times higher on the natural substrate cellobiose while both W127F and L178E mutants showed an enhancement in thermostability. The kinetic stability of W127F, V176A, L178A and L178E also increased between 2- and 3-folds compared to WT. Our results indicate that while the structure between subsites +1 and +2 is critical for the glucose tolerance, the specific residues may not be identical across such enzymes. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  4. Relationships between postweaning residual feed intake in heifers and forage use, body composition, feeding behavior, physical activity, and heart rate of pregnant beef females.

    Science.gov (United States)

    Hafla, A N; Carstens, G E; Forbes, T D A; Tedeschi, L O; Bailey, J C; Walter, J T; Johnson, J R

    2013-11-01

    The objectives of this study were to determine if residual feed intake (RFI) classification of beef heifers affected efficiency of forage utilization, body composition, feeding behavior, heart rate, and physical activity of pregnant females. Residual feed intake was measured in growing Bonsmara heifers for 2 yr (n=62 and 53/yr), and heifers with the lowest (n=12/yr) and highest (n=12/yr) RFI were retained for breeding. Of the 48 heifers identified as having divergent RFI, 19 second-parity and 23 first-parity females were used in the subsequent pregnant-female trial. Pregnant females were fed a chopped hay diet (ME=2.11 Mcal kg(-1) DM) in separate pens equipped with GrowSafe bunks to measure individual intake and feeding behavior. Body weights were measured at 7-d intervals and BCS and ultrasound measurements of 12th-rib fat depth, rump fat depth, and LM area obtained on d 0 and 77. Heart rate and physical activity were measured for 7 consecutive d. First-parity females had lower (Pgain, and initial hip height and tended (P=0.07) to have lower DMI compared to second-parity females. Females with low RFI as heifers consumed 17% less (Pheifers but maintained the same BW, BW gain, and body composition. Likewise, RFI classification did not affect calving date. An interaction (P=0.04) between heifer RFI classification and parity was found for calf birth weight. Calves from first-parity low-RFI females were lighter at birth (Pdaily step counts and greater lying-bout frequencies compared to second-parity females, but physical activity was not affected by RFI classification. Heart rates of females classified as low RFI were 7% lower (P=0.03) compared to high-RFI females. Heifer postweaning RFI but not G:F or residual gain were positively correlated with forage intake (r=0.38) and RFI (r=0.42) of pregnant females. Results indicate that heifers identified as having low postweaning RFI have greater efficiency of forage utilization as pregnant females, with minimal impacts on

  5. Residual limb wounds or ulcers heal in transtibial amputees using an active suction socket system. A randomized controlled study.

    Science.gov (United States)

    Traballesi, M; Delussu, A S; Fusco, A; Iosa, M; Averna, T; Pellegrini, R; Brunelli, S

    2012-12-01

    The factors that determine successful rehabilitation after lower limb amputation have been widely investigated in the literature, but little attention has been paid to the type of prosthesis and clinical state of the residual limb, particularly the presence of open ulcers. The aim of this study was to investigate the effects of a vacuum-assisted socket system (VASS) in a sample of trans-tibial amputees with wounds or ulcers on the stump and to evaluate prosthesis use as a primary outcome. Secondary outcome measures were mobility with the prosthesis, pain associated with prosthesis use, and wound/ulcer healing. Randomized controlled study. Inpatient. Twenty dysvascular transtibial amputees suffering from ulcers due to prosthesis use or delayed wound healing post-amputation were enrolled. Participants were separated into two groups: the experimental group was trained to use a VASS prosthesis in the presence of open ulcers/wounds on the stump; and the control group was trained to use a standard suction socket system prosthesis following ulcers/wounds healing. At the end of the 12-week rehabilitation program, all VASS users were able to walk independently with their prosthesis as reflected by a median Locomotor Capability Index (LCI) value of 42, whereas only five participants in the control group were able to walk independently with a median LCI value of 21. At the two-month follow-up, the participants used their VASS prostheses for 62 hours a week (median; range: 0-91), which was significantly longer than the control group using the standard prosthesis for 5 hours per week (range: 0-56, p=0.003). At the six-month follow-up, the difference between VASS-users (80, range: 0-112 hours a weeks) and control-users (59, range: 0-91) was no longer significant (p=0.191). Despite more intense use of the prosthesis, pain and wound healing did not significantly differ between the two groups. These results showed that the VASS prosthesis allowed early fitting with prompt

  6. Sharing Residual Liability

    DEFF Research Database (Denmark)

    Carbonara, Emanuela; Guerra, Alice; Parisi, Francesco

    2016-01-01

    Economic models of tort law evaluate the efficiency of liability rules in terms of care and activity levels. A liability regime is optimal when it creates incentives to maximize the value of risky activities net of accident and precaution costs. The allocation of primary and residual liability...... the virtues and limits of loss-sharing rules in generating optimal (second-best) incentives and allocations of risk. We find that loss sharing may be optimal in the presence of countervailing policy objectives, homogeneous risk avoiders, and subadditive risk, which potentially offers a valuable tool...

  7. The determination of pesticide residues and contaminants in vegetables by means of neutron activation analysis. Part of a coordinated programme on nuclear detection and analysis

    International Nuclear Information System (INIS)

    Mongkolphanta, S.

    1977-11-01

    Neutron activation techniques have been applied to study the existence of pesticide residues (As, Br, Hg) and pollutants (Cd, Co, Zn) in local vegetable and fruit samples, polished and unpolished glutinous and non-glutinous milled rice, and fish caught in polluted and unpolluted areas of the Gulf of Thailand. Since the baseline levels of these toxic elements in vegetables, fruits, rice and fish are not known as yet, the results cannot be regarded as conclusive. Furthermore, no significant difference was observed in the concentration of these elements in fish caught in polluted and unpolluted areas. The Gulf of Thailand does not therefore appear to be polluted at present. The substoichiometric isotope dilution technique was successfully applied to mercury determinations, and only to a limited extent to Cd due to shutdown of the TRR-1 reactor

  8. Triazacyclophane (TAC)-scaffolded histidine and aspartic acid residues as mimics of non-heme metalloenzyme active sites

    NARCIS (Netherlands)

    Albada, H.B.; Soulimani, F.; Jacobs, H.J.F.; Versluis, C.; Weckhuysen, B.M.; Liskamp, R.M.J.

    2012-01-01

    We describe the synthesis and coordination behaviour to copper(II) of two close structural triazacyclophane-based mimics of two often encountered aspartic acid and histidine containing metalloenzyme active sites. Coordination of these mimics to copper(I) and their reaction with molecular oxygen

  9. Antigen-driven CD4+ T cell and HIV-1 dynamics: residual viral replication under highly active antiretroviral therapy

    NARCIS (Netherlands)

    Ferguson, N. M.; DeWolf, F.; Ghani, A. C.; Fraser, C.; Donnelly, C. A.; Reiss, P.; Lange, J. M.; Danner, S. A.; Garnett, G. P.; Goudsmit, J.; Anderson, R. M.

    1999-01-01

    Antigen-induced stimulation of the immune system can generate heterogeneity in CD4+ T cell division rates capable of explaining the temporal patterns seen in the decay of HIV-1 plasma RNA levels during highly active antiretroviral therapy. Posttreatment increases in peripheral CD4+ T cell counts are

  10. Biochemical characterization of mutants in the active site residues of the beta-galactosidase enzyme of Bacillus circulans ATCC 31382

    NARCIS (Netherlands)

    Bultema, Jelle B.; Kuipers, Bas J. H.; Dijkhuizen, Lubbert

    2014-01-01

    The Bacillus circulans ATCC 31382 beta-galactosidase (BgaD) is a retaining-type glycosidase of glycoside hydrolase family 2 (GH2). Its commercial enzyme preparation, Biolacta N5, is used for commercial-scale production of galacto-oligosaccharides (GOS). The BgaD active site and catalytic amino acid

  11. Structure and Active Stie Residues of Pg1D, an N-Acetyltransferase from the Bacillosamine Synthetic Pathway Required for N-Glycan Synthesis in Campylobacter jejuni

    Energy Technology Data Exchange (ETDEWEB)

    Rangarajan,E.; Ruane, K.; Sulea, T.; Watson, D.; Proteau, A.; Leclerc, S.; Cygler, M.; Matte, A.; Young, N.

    2008-01-01

    Campylobacter jejuni is highly unusual among bacteria in forming N-linked glycoproteins. The heptasaccharide produced by its pgl system is attached to protein Asn through its terminal 2, 4-diacetamido-2, 4,6-trideoxy-d-Glc (QuiNAc4NAc or N, N'-diacetylbacillosamine) moiety. The crucial, last part of this sugar's synthesis is the acetylation of UDP-2-acetamido-4-amino-2, 4,6-trideoxy-d-Glc by the enzyme PglD, with acetyl-CoA as a cosubstrate. We have determined the crystal structures of PglD in CoA-bound and unbound forms, refined to 1.8 and 1.75 Angstroms resolution, respectively. PglD is a trimer of subunits each comprised of two domains, an N-terminal {alpha}/{beta}-domain and a C-terminal left-handed {beta}-helix. Few structural differences accompany CoA binding, except in the C-terminal region following the {beta}-helix (residues 189-195), which adopts an extended structure in the unbound form and folds to extend the {beta}-helix upon binding CoA. Computational molecular docking suggests a different mode of nucleotide-sugar binding with respect to the acetyl-CoA donor, with the molecules arranged in an 'L-shape', compared with the 'in-line' orientation in related enzymes. Modeling indicates that the oxyanion intermediate would be stabilized by the NH group of Gly143', with His125' the most likely residue to function as a general base, removing H+ from the amino group prior to nucleophilic attack at the carbonyl carbon of acetyl-CoA. Site-specific mutations of active site residues confirmed the importance of His125', Glu124', and Asn118. We conclude that Asn118 exerts its function by stabilizing the intricate hydrogen bonding network within the active site and that Glu124' may function to increase the pKa of the putative general base, His125'.

  12. Construction of a catalytically inactive cholesterol oxidase mutant: investigation of the interplay between active site-residues glutamate 361 and histidine 447.

    Science.gov (United States)

    Yin, Ye; Liu, Pingsheng; Anderson, Richard G W; Sampson, Nicole S

    2002-06-15

    Cholesterol oxidase catalyzes the oxidation of cholesterol to cholest-5-en-3-one and its subsequent isomerization into cholest-4-en-3-one. Two active-site residues, His447 and Glu361, are important for catalyzing the oxidation and isomerization reactions, respectively. Double-mutants were constructed to test the interplay between these residues in catalysis. We observed that the k(cat) of oxidation for the H447Q/E361Q mutant was 3-fold less than that for H447Q and that the k(cat) of oxidation for the H447E/E361Q mutant was 10-fold slower than that for H447E. Because both doubles-mutants do not have a carboxylate at position 361, they do not catalyze isomerization of the reaction intermediate cholest-5-en-3-one to cholest-4-en-3-one. These results suggest that Glu361 can compensate for the loss of histidine at position 447 by acting as a general base catalyst for oxidation of cholesterol. Importantly, the construction of the double-mutant H447E/E361Q yields an enzyme that is 31,000-fold slower than wild type in k(cat) for oxidation. The H447E/E361Q mutant is folded like native enzyme and still associates with model membranes. Thus, this mutant may be used to study the effects of membrane binding in the absence of catalytic activity. It is demonstrated that in assays with caveolae membrane fractions, the wild-type enzyme uncouples platelet-derived growth factor receptor beta (PDGFRbeta) autophosphorylation from tyrosine phosphorylation of neighboring proteins, and the H447E/E361Q mutant does not. Thus maintenance of membrane structure by cholesterol is important for PDGFRbeta-mediated signaling. The cholesterol oxidase mutant probe described will be generally useful for investigating the role of membrane structure in signal transduction pathways in addition to the PDGFRbeta-dependent pathway tested.

  13. Comparison of volumetric and functional parameters in simultaneous cardiac PET/MR: feasibility of volumetric assessment with residual activity from prior PET/CT

    Energy Technology Data Exchange (ETDEWEB)

    Luecke, C.; Brenneis, B.; Grothoff, M.; Gutberlet, M. [University Leipzig - Heart Center, Department of Diagnostic and Interventional Radiology, Leipzig (Germany); Oppolzer, B.; Werner, P.; Jochimsen, T.; Sattler, B.; Barthel, H.; Sabri, O. [University Hospital Leipzig, Department of Nuclear Medicine, Leipzig (Germany); Foldyna, B. [University Leipzig - Heart Center, Department of Diagnostic and Interventional Radiology, Leipzig (Germany); Massachusetts General Hospital - Harvard Medical School, Cardiac MR PET CT Program, Boston, MA (United States); Lurz, P. [University Leipzig - Heart Center, Clinic for Internal Medicine/Cardiology, Leipzig (Germany); Lehmkuhl, L. [Herz- und Gefaess-Klinik GmbH, Radiologische Klinik, Bad Neustadt (Germany)

    2017-12-15

    To compare cardiac left ventricular (LV) parameters in simultaneously acquired hybrid fluorine-18-fluorodeoxyglucose ([18F] FDG) positron emission tomography/magnetic resonance imaging (PET/MRI) in patients with residual tracer activity of upstream PET/CT. Twenty-nine patients (23 men, age 58±17 years) underwent cardiac PET/MRI either directly after a non-cardiac PET/CT with homogenous cardiac [18F] FDG uptake (n=20) or for viability assessment (n=9). Gated cardiac [18F] FDG PET and cine MR sequences were acquired simultaneously and evaluated blinded to the cross-imaging results. Image quality (IQ), end-diastolic (LVEDV), end-systolic volume (LVESV), ejection fraction (LVEF) and myocardial mass (LVMM) were measured. Pearson correlation and intraclass correlation coefficient (ICC), regression and a Bland-Altman analysis were assessed. Except LVMM, volumetric and functional LV parameters demonstrated high correlations (LVESV: r=0.97, LVEDV: r=0.95, LVEF: r=0.91, LVMM: r=0.87, each p<0.05), but wide limits of agreement (LOA) for LVEDV (-25.3-82.5ml); LVESV (-33.1-72.7ml); LVEF (-18.9-14.8%) and LVMM (-78.2-43.2g). Intra- and interobserver reliability were very high (ICC≥0.95) for all parameters, except for MR-LVEF (ICC=0.87). PET-IQ (0-3) was high (mean: 2.2±0.9) with significant influence on LVMM calculations only. In simultaneously acquired cardiac PET/MRI data, LVEDV, LVESV and LVEF show good agreement. However, the agreement seems to be limited if cardiac PET/MRI follows PET/CT and only the residual activity is used. (orig.)

  14. A Functional Interplay between Human Immunodeficiency Virus Type 1 Protease Residues 77 and 93 Involved in Differential Regulation of Precursor Autoprocessing and Mature Protease Activity.

    Directory of Open Access Journals (Sweden)

    Christopher J Counts

    Full Text Available HIV-1 protease (PR is a viral enzyme vital to the production of infectious virions. It is initially synthesized as part of the Gag-Pol polyprotein precursor in the infected cell. The free mature PR is liberated as a result of precursor autoprocessing upon virion release. We previously described a model system to examine autoprocessing in transfected mammalian cells. Here, we report that a covariance analysis of miniprecursor (p6*-PR sequences derived from drug naïve patients identified a series of amino acid pairs that vary together across independent viral isolates. These covariance pairs were used to build the first topology map of the miniprecursor that suggests high levels of interaction between the p6* peptide and the mature PR. Additionally, several PR-PR covariance pairs are located far from each other (>12 Å Cα to Cα relative to their positions in the mature PR structure. Biochemical characterization of one such covariance pair (77-93 revealed that each residue shows distinct preference for one of three alkyl amino acids (V, I, and L and that a polar or charged amino acid at either of these two positions abolishes precursor autoprocessing. The most commonly observed 77V is preferred by the most commonly observed 93I, but the 77I variant is preferred by other 93 variances (L, V, or M in supporting precursor autoprocessing. Furthermore, the 77I93V covariant enhanced precursor autoprocessing and Gag polyprotein processing but decreased the mature PR activity. Therefore, both covariance and biochemical analyses support a functional association between residues 77 and 93, which are spatially distant from each other in the mature PR structure. Our data also suggests that these covariance pairs differentially regulate precursor autoprocessing and the mature protease activity.

  15. Automated multi-filtration cleanup with nitrogen-enriched activated carbon material as pesticide multi-residue analysis method in representative crop commodities.

    Science.gov (United States)

    Qin, Yuhong; Zhang, Jingru; Li, Yifan; Wang, Qiuxiao; Wu, Yangliu; Xu, Lanshu; Jin, Xiaojuan; Pan, Canping

    2017-09-15

    An automated multi-filtration cleanup (Auto m-FC) method with nitrogen-enriched activated carbon material based on modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) extracts was developed. It was applied to pesticide multi-residue analysis in six representative crop commodities. The automatic device was aimed to improve the cleanup efficiency and reduce manual operation workload in cleanup step. By controlling extracts volume, flow rate and Auto m-FC cycles, the device could finish cleanup process accurately. In this work, nitrogen-enriched activated carbon mixed with alternative sorbents and anhydrous magnesium sulfate (MgSO 4 ) was packed in a column for Auto m-FC and followed by liquid chromatography with tandem mass spectrometric (LC-MS/MS) detection. This newly developed carbon material showed excellent cleanup performance. It was validated by analyzing 23 pesticides in six representative matrices spiked at two concentration levels of 10 and 100μg/kg. Water addition volume, salts, sorbents, Auto m-FC procedure including the flow rate and the Auto m-FC cycles for each matrix were optimized. Then, three general Auto m-FC methods were introduced to high water content, high oil and starch content, difficult commodities. Spike recoveries were within 82 and 106% and 1-14% RSD for all analytes in the tested matrices. Matrix-matched calibrations were performed with the coefficients of determination over 0.997 between concentration levels of 10 and 1000μg/kg. The developed method was successfully applied to the determination of pesticide residues in market samples. Copyright © 2017. Published by Elsevier B.V.

  16. Productivity, Physicochemical Changes, and Antioxidant Activity of Shiitake Culinary-Medicinal Mushroom Lentinus edodes (Agaricomycetes) Cultivated on Lignocellulosic Residues.

    Science.gov (United States)

    Gaitán-Hernández, Rigoberto; Zavaleta, Marco Antonio Barradas; Aquino-Bolaños, Elia Nora

    2017-01-01

    The effects of substrate and strain on productivity, physicochemical characteristics, and compounds with antioxidant activity were evaluated in basidiomes of the shiitake mushroom, Lentinus edodes. Strains IE-245 and IE-256 and the substrates oak wood shavings (OW), sorghum stubble (SS), and sugar cane bagasse (SC) were used. Productivity was evaluated by measuring biological efficiency (BE), production rate (PR), and yield. Total sugars, total soluble solids, pH, titratable acidity, color parameters, total phenolics, flavonoids, ascorbic acid, and antioxidant activity of the basidiomes were measured. BE, PR and yield were higher with the combination IE-256/SS, at 103.71%, 1.32%, and 34.57%, respectively. The largest amount of total sugars (17.61 mg glucose · g-1 dry weight) was found with combination IE-256/SS. Variation was observed in basidiome color; the lowest luminosity (L*) value (darkest color) was found in the IE-256 strain on the OW substrate (L* = 30.45), whereas that of the IE-245 strain on the SC substrate was the lightest in color (L* = 57.00). The largest amounts of total phenolics were recorded in the IE-256 strain on the OW (6.50 mg gallic acid equivalents [GAE] · g-1 dry weight) and the SS substrates (5.85 mg GAE · g-1 dry weight). The best antioxidant activity was obtained with IE-256-0.80, 0.65, and 0.59 μmol Trolox equivalents · g dry weight-1-on the OW, SC, and SS substrates, respectively. Based on the values of BE, PR, and yield, IE-256/SS was the most productive. Substrate and strain, and their interactions, influenced the physicochemical characteristics of the basidiomes and the amounts of compounds with antioxidant activity they contained.

  17. Site directed mutagenesis of amino acid residues at the active site of mouse aldehyde oxidase AOX1.

    Directory of Open Access Journals (Sweden)

    Silvia Schumann

    Full Text Available Mouse aldehyde oxidase (mAOX1 forms a homodimer and belongs to the xanthine oxidase family of molybdoenzymes which are characterized by an essential equatorial sulfur ligand coordinated to the molybdenum atom. In general, mammalian AOs are characterized by broad substrate specificity and an yet obscure physiological function. To define the physiological substrates and the enzymatic characteristics of mAOX1, we established a system for the heterologous expression of the enzyme in Escherichia coli. The recombinant protein showed spectral features and a range of substrate specificity similar to the native protein purified from mouse liver. The EPR data of recombinant mAOX1 were similar to those of AO from rabbit liver, but differed from the homologous xanthine oxidoreductase enzymes. Site-directed mutagenesis of amino acids Val806, Met884 and Glu1265 at the active site resulted in a drastic decrease in the oxidation of aldehydes with no increase in the oxidation of purine substrates. The double mutant V806E/M884R and the single mutant E1265Q were catalytically inactive enzymes regardless of the aldehyde or purine substrates tested. Our results show that only Glu1265 is essential for the catalytic activity by initiating the base-catalyzed mechanism of substrate oxidation. In addition, it is concluded that the substrate specificity of molybdo-flavoenzymes is more complex and not only defined by the three characterized amino acids in the active site.

  18. Tak v současných mediálních mluvených rozhovorech: ty zákony co se vztahujou teďkon na nás soukromníky (2. část)

    Czech Academy of Sciences Publication Activity Database

    Jílková, Lucie

    -, č. 15 (2017), s. 31-46 ISSN 1804-137X R&D Projects: GA ČR GA15-01116S Institutional support: RVO:68378092 Keywords : word tak * DIALOG corpus * mass media dialogue * spoken language * meanings and functions of the Czech word tak Subject RIV: AI - Linguistics OBOR OECD: Linguistics

  19. Effect of ultrasonic extraction conditions on antioxidative and immunomodulatory activities of a Ganoderma lucidum polysaccharide originated from fermented soybean curd residue.

    Science.gov (United States)

    Shi, Min; Yang, Yingnan; Hu, Xuansheng; Zhang, Zhenya

    2014-07-15

    A crude Ganoderma lucidum polysaccharide (GLPL) was extracted from fermented soybean curd residue by ultrasonic assisted extraction. The optimal extraction conditions were 30 min at 80 °C with 80 W and water to solid ratio of 10, and with this method 115.47 ± 2.95 mg/g of GLPL yield was obtained. Additionally, the antioxidant and immunomodulatory activities of GLPL were investigated. The results showed that GLPL exhibited strong antioxidant effects, which included scavenging activities against DPPH radicals, hydrogen oxide and ABTS radicals with IC50 values of 0.23, 0.48 and 0.69 mg/mL, respectively. For immunomodulatory activities, GLPL was shown to strongly stimulate the proliferation of macrophages (158.02 ± 13.12%), the production of nitric oxide and phagocytosis (21.16 ± 1.65 μM), and, at 40.00 μg/mL, protected macrophage from Doxorubicin (DOX) (0.16 ± 0.003). Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Production of bio-oil rich in acetic acid and phenol from fast pyrolysis of palm residues using a fluidized bed reactor: Influence of activated carbons.

    Science.gov (United States)

    Jeong, Jae-Yong; Lee, Uen-Do; Chang, Won-Seok; Jeong, Soo-Hwa

    2016-11-01

    In this study, palm residues were pyrolyzed in a bench-scale (3kg/h) fast pyrolysis plant equipped with a fluidized bed reactor and bio-oil separation system for the production of bio-oil rich in acetic acid and phenol. Pyrolysis experiments were performed to investigate the effects of reaction temperature and the types and amounts of activated carbon on the bio-oil composition. The maximum bio-oil yield obtained was approximately 47wt% at a reaction temperature of 515°C. The main compounds produced from the bio-oils were acetic acid, hydroxyacetone, phenol, and phenolic compounds such as cresol, xylenol, and pyrocatechol. When coal-derived activated carbon was applied, the acetic acid and phenol yields in the bio-oils reached 21 and 19wt%, respectively. Finally, bio-oils rich in acetic acid and phenol could be produced separately by using an in situ bio-oil separation system and activated carbon as an additive. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Transforming growth factor β-activated kinase 1 negatively regulates interleukin-1α-induced stromal-derived factor-1 expression in vascular smooth muscle cells

    International Nuclear Information System (INIS)

    Yang, Bin; Li, Wei; Zheng, Qichang; Qin, Tao; Wang, Kun; Li, Jinjin; Guo, Bing; Yu, Qihong; Wu, Yuzhe; Gao, Yang; Cheng, Xiang; Hu, Shaobo; Kumar, Stanley Naveen; Liu, Sanguang; Song, Zifang

    2015-01-01

    Stromal-derived Factor-1 (SDF-1) derived from vascular smooth muscle cells (VSMCs) contributes to vascular repair and remodeling in various vascular diseases. In this study, the mechanism underlying regulation of SDF-1 expression by interleukin-1α (IL-1α) was investigated in primary rat VSMCs. We found IL-1α promotes SDF-1 expression by up-regulating CCAAT-enhancer-binding protein β (C/EBPβ) in an IκB kinase β (IKKβ) signaling-dependent manner. Moreover, IL-1α-induced expression of C/EBPβ and SDF-1 was significantly potentiated by knockdown of transforming growth factor β-activated kinase 1 (TAK1), an upstream activator of IKKβ signaling. In addition, we also demonstrated that TAK1/p38 mitogen-activated protein kinase (p38 MAPK) signaling exerted negative effect on IL-1α-induced expression of C/EBPβ and SDF-1 through counteracting ROS-dependent up-regulation of nuclear factor erythroid 2-related factor 2 (NRF2). In conclusion, TAK1 acts as an important regulator of IL-1α-induced SDF-1 expression in VSMCs, and modulating activity of TAK1 may serve as a potential strategy for modulating vascular repair and remodeling. - Highlights: • IL-1α induces IKKβ signaling-dependent SDF-1 expression by up-regulating C/EBPβ. • Activation of TAK1 by IL-1α negatively regulates C/EBPβ-dependent SDF-1 expression. • IL-1α-induced TAK1/p38 MAPK signaling counteracts ROS-dependent SDF-1 expression. • TAK1 counteracts IL-1α-induced SDF-1 expression by attenuating NRF2 up-regulation

  2. Transforming growth factor β-activated kinase 1 negatively regulates interleukin-1α-induced stromal-derived factor-1 expression in vascular smooth muscle cells

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Bin [Department of Hepatobiliary Surgery, Union Hospital, Tongji Medical College, Huangzhong University of Science and Technology, Wuhan 430022 (China); Li, Wei [Department of Gerontology, Union Hospital, Tongji Medical College, Huangzhong University of Science and Technology, Wuhan 430022 (China); Zheng, Qichang [Department of Hepatobiliary Surgery, Union Hospital, Tongji Medical College, Huangzhong University of Science and Technology, Wuhan 430022 (China); Qin, Tao [Department of Hepatobiliary Pancreatic Surgery, People' s Hospital of Zhengzhou University, School of Medicine, Zhengzhou University, Zhengzhou 450003 (China); Wang, Kun; Li, Jinjin; Guo, Bing; Yu, Qihong; Wu, Yuzhe; Gao, Yang; Cheng, Xiang; Hu, Shaobo; Kumar, Stanley Naveen [Department of Hepatobiliary Surgery, Union Hospital, Tongji Medical College, Huangzhong University of Science and Technology, Wuhan 430022 (China); Liu, Sanguang, E-mail: sanguang1998@sina.com [Department of Hepatobiliary Surgery, The Second Hospital, Hebei Medical University, Shijiazhuang 050000 (China); Song, Zifang, E-mail: zsong@hust.edu.cn [Department of Hepatobiliary Surgery, Union Hospital, Tongji Medical College, Huangzhong University of Science and Technology, Wuhan 430022 (China)

    2015-07-17

    Stromal-derived Factor-1 (SDF-1) derived from vascular smooth muscle cells (VSMCs) contributes to vascular repair and remodeling in various vascular diseases. In this study, the mechanism underlying regulation of SDF-1 expression by interleukin-1α (IL-1α) was investigated in primary rat VSMCs. We found IL-1α promotes SDF-1 expression by up-regulating CCAAT-enhancer-binding protein β (C/EBPβ) in an IκB kinase β (IKKβ) signaling-dependent manner. Moreover, IL-1α-induced expression of C/EBPβ and SDF-1 was significantly potentiated by knockdown of transforming growth factor β-activated kinase 1 (TAK1), an upstream activator of IKKβ signaling. In addition, we also demonstrated that TAK1/p38 mitogen-activated protein kinase (p38 MAPK) signaling exerted negative effect on IL-1α-induced expression of C/EBPβ and SDF-1 through counteracting ROS-dependent up-regulation of nuclear factor erythroid 2-related factor 2 (NRF2). In conclusion, TAK1 acts as an important regulator of IL-1α-induced SDF-1 expression in VSMCs, and modulating activity of TAK1 may serve as a potential strategy for modulating vascular repair and remodeling. - Highlights: • IL-1α induces IKKβ signaling-dependent SDF-1 expression by up-regulating C/EBPβ. • Activation of TAK1 by IL-1α negatively regulates C/EBPβ-dependent SDF-1 expression. • IL-1α-induced TAK1/p38 MAPK signaling counteracts ROS-dependent SDF-1 expression. • TAK1 counteracts IL-1α-induced SDF-1 expression by attenuating NRF2 up-regulation.

  3. HEU Measurements of Holdup and Recovered Residue in the Deactivation and Decommissioning Activities of the 321-M Reactor Fuel Fabrication Facility at the Savannah River Site

    Energy Technology Data Exchange (ETDEWEB)

    DEWBERRY, RAYMOND; SALAYMEH, SALEEM R.; CASELLA, VITO R.; MOORE, FRANK S.

    2005-03-11

    This paper contains a summary of the holdup and material control and accountability (MC&A) assays conducted for the determination of highly enriched uranium (HEU) in the deactivation and decommissioning (D&D) of Building 321-M at the Savannah River Site (SRS). The 321-M facility was the Reactor Fuel Fabrication Facility at SRS and was used to fabricate HEU fuel assemblies, lithium-aluminum target tubes, neptunium assemblies, and miscellaneous components for the SRS production reactors. The facility operated for more than 35 years. During this time thousands of uranium-aluminum-alloy (U-Al) production reactor fuel tubes were produced. After the facility ceased operations in 1995, all of the easily accessible U-Al was removed from the building, and only residual amounts remained. The bulk of this residue was located in the equipment that generated and handled small U-Al particles and in the exhaust systems for this equipment (e.g., Chip compactor, casting furnaces, log saw, lathes A & B, cyclone separator, Freon{trademark} cart, riser crusher, ...etc). The D&D project is likely to represent an important example for D&D activities across SRS and across the Department of Energy weapons complex. The Savannah River National Laboratory was tasked to conduct holdup assays to quantify the amount of HEU on all components removed from the facility prior to placing in solid waste containers. The U-235 holdup in any single component of process equipment must not exceed 50 g in order to meet the container limit. This limit was imposed to meet criticality requirements of the low level solid waste storage vaults. Thus the holdup measurements were used as guidance to determine if further decontamination of equipment was needed to ensure that the quantity of U-235 did not exceed the 50 g limit and to ensure that the waste met the Waste Acceptance Criteria (WAC) of the solid waste storage vaults. Since HEU is an accountable nuclear material, the holdup assays and assays of recovered

  4. Effect of alterations of key active site residues in O6-alkylguanine-DNA Alkyltransferase on its ability to modulate the genotoxicity of 1,2-dibromoethane.

    Science.gov (United States)

    Liu, Liping; Watanabe, Kengo; Fang, Qingming; Williams, Kevin M; Guengerich, F Peter; Pegg, Anthony E

    2007-01-01

    The production of mutations and the reduction in survival of cells treated with alpha,omega-dihaloalkanes is greatly enhanced by the presence of O6-alkylguanine-DNA alkyltransferase (AGT), a DNA repair protein that removes O6-alkylguanine adducts from DNA [Liu, L., Hachey, D. L., Valadez, G., Williams, K. M., Guengerich, F. P., Loktionova, N. A., Kanugula, S., and Pegg, A. E. (2004) J. Biol. Chem. 279, 4250-4259]. The effects of alterations to key residues in the active site of AGT were studied using AGTs with point mutations. It was found that mutants of AGT at positions Tyr114, Arg128, Pro140, Gly156, Gly160, and Tyr158 did not bring about the increase in genotoxicity of 1,2-dibromoethane seen with wild-type AGT, although these mutants, with the exception of those at Tyr114 and Arg128, are known to have sufficient AGT repair function to be able to protect cells from alkylating agents. The R128A mutant was able to react with 1,2-dibromoethane at the Cys145 acceptor site, but the resulting AGT-Cys145S-(CH2)2Br was much less able to produce a covalent adduct with DNA. This result is explained by the need for AGT to induce a structural change in the DNA "flipping" of a guanine nucleotide into the substrate binding pocket where Cys145 is located since the side chain of residue Arg128 plays a critical role in this reaction. Point mutations in AGT at the other sites (Y114A, P140K, and Y158H) reduced the ability of the protein to react with 1,2-dibromoethane as measured by the loss of activity. These results were confirmed by MS analysis of the tryptic peptide that contains the modified Cys145. There was no change in the stability of the AGT-Cys145S-(CH2)2Br intermediate formed in mutants Y158H and P140K. The reaction was studied in detail with mutant P140K using dihaloalkanes of different length; no effect of the mutations was seen with dibromomethane, but an enhanced difference was observed with 1,3-dibromopropane and 1,5-dibromopentane. These results show that even

  5. Design of an Anticancer Copper(II) Prodrug Based on the Lys199 Residue of the Active Targeting Human Serum Albumin Nanoparticle Carrier.

    Science.gov (United States)

    Gou, Yi; Zhang, Yao; Zhang, Zhenlei; Wang, Jun; Zhou, Zuping; Liang, Hong; Yang, Feng

    2017-06-05

    We not only modified the types and numbers of coordinated ligands in a metal agent to enhance its anticancer activity, but we also designed a metal prodrug based on the N-donor residues of the human serum albumin (HSA) IIA subdomain to improve its delivery efficiency and selectivity in vivo. However, there may be a conflict in simultaneously achieving the two goals because Lys199 and His242 in the IIA subdomain of HSA can replace its two coordinated ligands, which will decrease its anticancer activity relative to the original metal agent. Thus, to improve the delivery efficiency of the metal agent and simultaneously avoid decreasing its anticancer activity in vivo, we decided to develop an anticancer metal prodrug by regulating its pharmacophore ligand so that it would not be displaced by the Lys199 residue of the folic acid (FA)-functionalized HSA nanoparticle (NP) carrier. To this end, we first synthesized two (E)-N'-(5-chloro-2-hydroxybenzylidene)benzohydrazide Schiff base (HL) Cu(II) compounds by designing a second ligand with a different coordinating atom with Cu 2+ /Cu(L)(QL)(Br) [C1, QL = quinolone] and Cu(L)(DMF)(Br) [C2, DMF = N,N-dimethylformamide]. As revealed by the structures of the two HSA complexes, the Cu compounds bind to the hydrophobic cavity in the HSA IIA subdomain. The QL ligand of C1 is replaced by Lys199, which coordinates with Cu 2+ , whereas the DMF ligand of C2 is kept intact and His242 is replaced with Br - of C2 and coordinates with Cu 2+ . The cytotoxicity of the Cu compounds was enhanced by the FA-HSA NPs in the Bel-7402 cells approximately 2-4-fold; however, they raise the cytotoxicity levels in the normal cells in vitro, and the FA-HSA NPs did not. Importantly, the in vivo data showed that FA-HSA-C2 NPs increased selectivity and the capacity to inhibit tumor growth and were less toxic than HSA-C2 NPs and C2. Moreover, C2/HSA-C2 NPs/FA-HSA-C2 NPs induced Bel-7402 cell death by potentially multiple mechanisms.

  6. Indoor residual spraying with a mixture of clothianidin (a neonicotinoid insecticide) and deltamethrin provides improved control and long residual activity against pyrethroid resistant Anopheles gambiae sl in Southern Benin.

    OpenAIRE

    Ngufor, C; Fongnikin, A; Rowland, M; N'Guessan, R

    2017-01-01

    There is an urgent need for new insecticides for indoor residual spraying (IRS) which can provide improved and prolonged control of malaria vectors that have developed resistance to existing insecticides. The neonicotinoid, clothianidin represents a class of chemistry new to public health. Clothianidin acts as an agonist on nicotinic acetyl choline receptors. IRS with a mixture of Clothianidin and another WHO approved insecticide such as deltamethrin could provide improved control of insectic...

  7. Kinetics of coffee industrial residue pyrolysis using distributed activation energy model and components separation of bio-oil by sequencing temperature-raising pyrolysis.

    Science.gov (United States)

    Chen, Nanwei; Ren, Jie; Ye, Ziwei; Xu, Qizhi; Liu, Jingyong; Sun, Shuiyu

    2016-12-01

    This study was carried out to investigate the kinetics of coffee industrial residue (CIR) pyrolysis, the effect of pyrolysis factors on yield of bio-oil component and components separation of bio-oil. The kinetics of CIR pyrolysis was analyzed using distributed activation energy model (DAEM), based on the experiments in thermogravimetric analyzer (TGA), and it indicated that the average of activation energy (E) is 187.86kJ·mol -1 . The bio-oils were prepared from CIR pyrolysis in vacuum tube furnace, and its components were determined by gas chromatography/mass spectrometry (GC-MS). Among pyrolysis factors, pyrolysis temperature is the most influential factor on components yield of bio-oil, directly concerned with the volatilization and yield of components (palmitic acid, linoleic acid, oleic acid, octadecanoic acid and caffeine). Furthermore, a new method (sequencing temperature-raising pyrolysis) was put forward and applied to the components separation of bio-oil. Based on experiments, a solution of components separation of bio-oil was come out. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Optimization of Ultrasonic-Assisted Enzymatic Extraction Conditions for Improving Total Phenolic Content, Antioxidant and Antitumor Activities In Vitro from Trapa quadrispinosa Roxb. Residues.

    Science.gov (United States)

    Li, Feng; Mao, Yi-Dan; Wang, Yi-Fan; Raza, Aun; Qiu, Li-Peng; Xu, Xiu-Quan

    2017-03-06

    Stems are the important residues of Trapa quadrispinosa Roxb., which are abundant in phenolic compounds. Ultrasonic-assisted enzymatic extraction (UAEE) is confirmed as a novel extraction technology with main advantages of enhancing extraction yield and physiological activities of the extracts from various plants. In this study, UAEE was applied to obtain the highest yield of phenolic content, strongest antioxidant, and antitumor activities and to optimize the extraction conditions using response surface methodology (RSM). The extracts from the stems of T. quadrispinosa were characterized by determination of their antioxidant activities through 2,2-azinobis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS), 1,1-Diphenyl-2-picrylhydrazxyl (DPPH) radical scavenging, total antioxidant capacity (TAC), ferric reducing antioxidant capacity (FRAC) methods and of their antitumor activity by MTT method. The selected key independent variables were cellulase concentration ( X ₁: 1.5%-2.5%), extraction time ( X ₂: 20-30 min) and extraction temperature ( X ₃: 40-60 °C). The optimal extraction conditions for total phenolic content (TPC) value of the extracts were determined as 1.74% cellulase concentration, 25.5 min ultrasonic extraction time and 49.0 °C ultrasonic temperature. Under these conditions, the highest TPC value of 53.6 ± 2.2 mg Gallic acid equivalent (GAE)/g dry weight (DW) was obtained, which agreed well with the predicted value (52.596 mg GAE/g·DW. Furthermore, the extracts obtained from UAEE presented highest antioxidant activities through ABTS, DPPH, TAC and FRAC methods were of 1.54 ± 0.09 mmol Trolox equivalent (TE)/g·DW; 1.45 ± 0.07 mmol·TE/g·DW; 45.2 ± 2.2 mg·GAE/g·DW; 50.4 ± 2.6 μmol FeSO₄ equivalent/g·DW and lowest IC 50 values of 160.4 ± 11.6 μg/mL, 126.1 ± 10.8 μg/mL, and 178.3 ± 13.1 μg/mL against Hela, HepG-2 and U251 tumor cells, respectively. The results indicated that the UAEE was an efficient alternative to improve

  9. Marine Tar Residues: a Review.

    Science.gov (United States)

    Warnock, April M; Hagen, Scott C; Passeri, Davina L

    Marine tar residues originate from natural and anthropogenic oil releases into the ocean environment and are formed after liquid petroleum is transformed by weathering, sedimentation, and other processes. Tar balls, tar mats, and tar patties are common examples of marine tar residues and can range in size from millimeters in diameter (tar balls) to several meters in length and width (tar mats). These residues can remain in the ocean environment indefinitely, decomposing or becoming buried in the sea floor. However, in many cases, they are transported ashore via currents and waves where they pose a concern to coastal recreation activities, the seafood industry and may have negative effects on wildlife. This review summarizes the current state of knowledge on marine tar residue formation, transport, degradation, and distribution. Methods of detection and removal of marine tar residues and their possible ecological effects are discussed, in addition to topics of marine tar research that warrant further investigation. Emphasis is placed on benthic tar residues, with a focus on the remnants of the Deepwater Horizon oil spill in particular, which are still affecting the northern Gulf of Mexico shores years after the leaking submarine well was capped.

  10. SIRT1 Regulates Thyroid-Stimulating Hormone Release by Enhancing PIP5Kgamma Activity through Deacetylation of Specific Lysine Residues in Mammals.

    Directory of Open Access Journals (Sweden)

    Sayaka Akieda-Asai

    Full Text Available BACKGROUND: SIRT1, a NAD-dependent deacetylase, has diverse roles in a variety of organs such as regulation of endocrine function and metabolism. However, it remains to be addressed how it regulates hormone release there. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report that SIRT1 is abundantly expressed in pituitary thyrotropes and regulates thyroid hormone secretion. Manipulation of SIRT1 level revealed that SIRT1 positively regulated the exocytosis of TSH-containing granules. Using LC/MS-based interactomics, phosphatidylinositol-4-phosphate 5-kinase (PIP5Kgamma was identified as a SIRT1 binding partner and deacetylation substrate. SIRT1 deacetylated two specific lysine residues (K265/K268 in PIP5Kgamma and enhanced PIP5Kgamma enzyme activity. SIRT1-mediated TSH secretion was abolished by PIP5Kgamma knockdown. SIRT1 knockdown decreased the levels of deacetylated PIP5Kgamma, PI(4,5P(2, and reduced the secretion of TSH from pituitary cells. These results were also observed in SIRT1-knockout mice. CONCLUSIONS/SIGNIFICANCE: Our findings indicated that the control of TSH release by the SIRT1-PIP5Kgamma pathway is important for regulating the metabolism of the whole body.

  11. Site-directed mutagenesis under the direction of in silico protein docking modeling reveals the active site residues of 3-ketosteroid-Δ1-dehydrogenase from Mycobacterium neoaurum.

    Science.gov (United States)

    Qin, Ning; Shen, Yanbing; Yang, Xu; Su, Liqiu; Tang, Rui; Li, Wei; Wang, Min

    2017-07-01

    3-Ketosteroid-Δ 1 -dehydrogenases (KsdD) from Mycobacterium neoaurum could transform androst-4-ene-3,17-dione (AD) to androst-1,4-diene-3,17-dione. This reaction has a significant effect on the product of pharmaceutical steroid. The crystal structure and active site residues information of KsdD from Mycobacterium is not yet available, which result in the engineering of KsdD is tedious. In this study, by the way of protein modeling and site-directed mutagenesis, we find that, Y122, Y125, S138, E140 and Y541 from the FAD-binding domain and Y365 from the catalytic domain play a key role in this transformation. Compared with the wild type, the decline in AD conversion for mutants illustrated that Y125, Y365, and Y541 were essential to the function of KsdD. Y122, S138 and E140 contributed to the catalysis of KsdD. The following analysis revealed the catalysis mechanism of these mutations in KsdD of Mycobacterium. These information presented here facilitate the manipulation of the catalytic properties of the enzyme to improve its application in the pharmaceutical steroid industry.

  12. Structure-based network analysis of activation mechanisms in the ErbB family of receptor tyrosine kinases: the regulatory spine residues are global mediators of structural stability and allosteric interactions.

    Directory of Open Access Journals (Sweden)

    Kevin A James

    Full Text Available The ErbB protein tyrosine kinases are among the most important cell signaling families and mutation-induced modulation of their activity is associated with diverse functions in biological networks and human disease. We have combined molecular dynamics simulations of the ErbB kinases with the protein structure network modeling to characterize the reorganization of the residue interaction networks during conformational equilibrium changes in the normal and oncogenic forms. Structural stability and network analyses have identified local communities integrated around high centrality sites that correspond to the regulatory spine residues. This analysis has provided a quantitative insight to the mechanism of mutation-induced "superacceptor" activity in oncogenic EGFR dimers. We have found that kinase activation may be determined by allosteric interactions between modules of structurally stable residues that synchronize the dynamics in the nucleotide binding site and the αC-helix with the collective motions of the integrating αF-helix and the substrate binding site. The results of this study have pointed to a central role of the conserved His-Arg-Asp (HRD motif in the catalytic loop and the Asp-Phe-Gly (DFG motif as key mediators of structural stability and allosteric communications in the ErbB kinases. We have determined that residues that are indispensable for kinase regulation and catalysis often corresponded to the high centrality nodes within the protein structure network and could be distinguished by their unique network signatures. The optimal communication pathways are also controlled by these nodes and may ensure efficient allosteric signaling in the functional kinase state. Structure-based network analysis has quantified subtle effects of ATP binding on conformational dynamics and stability of the EGFR structures. Consistent with the NMR studies, we have found that nucleotide-induced modulation of the residue interaction networks is not

  13. p.X654R IDUA variant among Thai individuals with intermediate mucopolysaccharidosis type I and its residual activity as demonstrated in COS-7 cells.

    Science.gov (United States)

    Ngiwsara, Lukana; Ketudat-Cairns, James R; Sawangareetrakul, Phannee; Charoenwattanasatien, Ratana; Champattanachai, Voraratt; Kuptanon, Chulaluck; Pangkanon, Suthipong; Tim-Aroon, Thipwimol; Wattanasirichaigoon, Duangrurdee; Svasti, Jisnuson

    2017-12-28

    Mucopolysaccharidosis type I (MPS I) is a rare autosomal-recessive disorder caused by defects in alpha-L-iduronidase (IDUA), a lysosomal enzyme encoded by the IDUA gene. Herein, we characterized IDUA mutations underlying mucopolysaccharidosis type I intermediate form (Hurler-Scheie syndrome) and its molecular pathogenic mechanisms. Clinical data, activity of the IDUA enzyme in leukocytes, and a mutation of the IDUA gene were analyzed. Pathogenesis associated with an IDUA mutation was further investigated by evaluating the mutant cDNA sequence, protein expression and activity in COS-7 cells. Five unrelated patients were identified to have clinical diagnosis of intermediate form of MPS I (Hurler-Scheie) and exhibited low-to-absent levels of leukocyte IDUA activity. Genetic analysis revealed homozygous c.*1T>C (p.X654R) mutation in two patients and compound heterozygosity between the c.*1T>C and another allele including c.265G>A (p.R89Q), c.935G>A (p.W312X), or c.1138 C>T (p.Q380X), each in a single patient. Sequencing the 3'RACE product of the c.*1T>C (p.X654R) allele indicated a 38-amino acids elongation of the mutant protein. COS-7 cells expressing IDUA with the mutations exhibited extremely low levels or complete absence of enzyme activity compared to wild-type IDUA. Western blot analysis detected no protein in p.W312X and p.Q380X samples, while an elevated molecular mass and a different pattern of protein bands were found in p.X654R specimen compared with the wild type IDUA. Mutational spectrum underlying intermediate MPS I is expanded. Our data suggest that the p.X654R is an intermediate IDUA mutant allele with residual enzyme activity. It can lead to intermediate or milder form of MPS I depending on another associated allele. © 2017 John Wiley & Sons Ltd/University College London.

  14. The dapE-encoded N-succinyl-L,L-Diaminopimelic Acid Desuccinylase from Haemophilus influenzae Contains two Active Site Histidine Residues

    Science.gov (United States)

    Gillner, Danuta M.; Bienvenue, David L.; Nocek, Boguslaw P.; Joachimiak, Andrzej; Zachary, Vincentos; Bennett, Brian; Holz, Richard C.

    2009-01-01

    The catalytic and structural properties of the H67A and H349A altered dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE) from H. influenzae were investigated. Based on sequence alignment with CPG2 both H67 and H349 were predicted to be Zn(II) ligands. Catalytic activity was observed for the H67A altered DapE enzyme which exhibited kcat = 1.5 ± 0.5 sec−1 and Km = 1.4 ± 0.3 mM. No catalytic activity was observed for H349A under the experimental conditions used. The EPR and electronic absorption data indicate that the Co(II) ion bound to H349A-DapE is analogous to WT DapE after the addition of a single Co(II) ion. The addition of one equivalent of Co(II) to H67A altered DapE provides spectra that are very different from the first Co(II) binding site of the WT enzyme, but similar to the second binding site. The EPR and electronic absorption data, in conjunction with the kinetic data, are consistent with the assignment of H67 and H349 as active site metal ligands for the DapE from H. influenzae. Furthermore, the data suggest that H67 is a ligand in the first metal binding site while H349 resides in the second metal binding site. A three-dimensional homology structure of the DapE from H. influenzae was generated using the X-ray crystal structure of the DapE from N. meningitidis as a template and superimposed on the structure of AAP. This homology structure confirms the assignment of H67 and H349 as active site ligands. The superimposition of the homology model of DapE with the dizinc(II) structure of AAP indicates that within 4.0 Å of the Zn(II) binding sites of AAP, all of the amino acid residues of DapE are nearly identical. PMID:18712420

  15. Transforming growth factor β activated kinase 1: a potential therapeutic target for rheumatic diseases.

    Science.gov (United States)

    Fechtner, Sabrina; Fox, David A; Ahmed, Salahuddin

    2017-07-01

    Pro-inflammatory cytokines such as IL-1β, IL-6 and TNF-α are central regulators of autoinflammatory diseases. While targeting these cytokines has proven to be a successful clinical strategy, the long-term challenges such as drug resistance, lack of efficacy and poor clinical outcomes in some patients are some of the limitations faced by these therapies. This has ignited strategies to reduce inflammation by potentially targeting a variety of molecules, including cell surface receptors, signalling proteins and/or transcription factors to minimize cytokine-induced inflammation and tissue injury. In this regard, transforming growth factor β activated kinase 1 (TAK1) is activated in the inflammatory signal transduction pathways in response to IL-1β, TNF-α or toll-like receptor stimulation. Because of its ideal position upstream of mitogen-activated protein kinases and the IκB kinase complex in signalling cascades, targeting TAK1 may be an attractive strategy for treating diseases characterized by chronic inflammation. Here, we discuss the emerging role of TAK1 in mediating the IL-1β, TNF-α and toll-like receptor mediated inflammatory responses in diseases such as RA, OA, gout and SS. We also review evidence suggesting that TAK1 inhibition may have potential therapeutic value. Finally, we focus on the current status of the development of TAK1 inhibitors and suggest further opportunities for testing TAK1 inhibitors in rheumatic diseases. © The Author 2016. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  16. PRODUCTION OF PASSION FRUIT SEEDLINGS WITH THE RESIDUE OF AGROINDUSTRIAL PROCESSING OF POTATOES AS A STRATEGY IN THE SUSTAINABILITY OF AGRICULTURAL FAMILY ACTIVITY

    Directory of Open Access Journals (Sweden)

    Leonardo Humberto Silva e Castro

    2012-12-01

    Full Text Available In order to contribute to family farming sustainable development and to the environmental management of the residue generated from potato processing by the company "Bem Brasil Alimentos LTDA" from Araxa, Minas Gerais, Brazil, an experiment was conducted at the Experimental Farm of Araxa (FEAX - EPAMIG between February to April of 2012, aiming to analyze passion fruit seedlings development from the use of different doses of this residue such as substrate, to reduce costs of raw materials. The experiment was conducted in randomized block design with six treatments, four repetitions and 20 plants per plot, being the treatments, blends of soil (latosol with levels of 10%, 20% and 30% of potato processing residue associated to chemical treatment and a witness. After 70 days of sowing, the height of air part (cm, root length (cm, number of leaves and total dry matter (g were evaluated. The best treatment was the M4 which used latosol + chemical treatment + 20% of residue. It was diagnosed that the use of agroindustrial potato processing residue, as part of the substrate, is a favorable alternative for passion fruit seedlings development. Thus, it gives a destination to this residue instead of discarding it into the environment, as well as decreases the costs of raw material that permit its correct ecological use, besides generating more income for family farmers.

  17. The dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase from Haemophilus influenzae contains two active-site histidine residues.

    Science.gov (United States)

    Gillner, Danuta M; Bienvenue, David L; Nocek, Boguslaw P; Joachimiak, Andrzej; Zachary, Vincentos; Bennett, Brian; Holz, Richard C

    2009-01-01

    The catalytic and structural properties of the H67A and H349A dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) from Haemophilus influenzae were investigated. On the basis of sequence alignment with the carboxypeptidase from Pseudomonas sp. strain RS-16, both H67 and H349 were predicted to be Zn(II) ligands. The H67A DapE enzyme exhibited a decreased catalytic efficiency (180-fold) compared with wild-type (WT) DapE towards N-succinyldiaminopimelic acid. No catalytic activity was observed for H349A under the experimental conditions used. The electronic paramagnetic resonance (EPR) and electronic absorption data indicate that the Co(II) ion bound to H349A-DapE is analogous to that of WT DapE after the addition of a single Co(II) ion. The addition of 1 equiv of Co(II) to H67A DapE provides spectra that are very different from those of the first Co(II) binding site of the WT enzyme, but that are similar to those of the second binding site. The EPR and electronic absorption data, in conjunction with the kinetic data, are consistent with the assignment of H67 and H349 as active-site metal ligands for the DapE from H. influenzae. Furthermore, the data suggest that H67 is a ligand in the first metal binding site, while H349 resides in the second metal binding site. A three-dimensional homology structure of the DapE from H. influenzae was generated using the X-ray crystal structure of the DapE from Neisseria meningitidis as a template and superimposed on the structure of the aminopeptidase from Aeromonas proteolytica (AAP). This homology structure confirms the assignment of H67 and H349 as active-site ligands. The superimposition of the homology model of DapE with the dizinc(II) structure of AAP indicates that within 4.0 A of the Zn(II) binding sites of AAP all of the amino acid residues of DapE are nearly identical.

  18. Characterization of a bifunctional PutA homologue from Bradyrhizobium japonicum and identification of an active site residue that modulates proline reduction of the flavin adenine dinucleotide cofactor.

    Science.gov (United States)

    Krishnan, Navasona; Becker, Donald F

    2005-06-28

    PutA is a bifunctional flavoenzyme in bacteria that catalyzes the four-electron oxidation of proline to glutamate. In certain prokaryotes such as Escherichia coli, PutA is also a transcriptional repressor of the proline utilization (put) genes and thus is trifunctional. In this work, we have begun to assess differences between bifunctional and trifunctional PutA enzymes by examining the PutA protein from Bradyrhizobium japonicum (BjPutA). Primary structure analysis of BjPutA shows it lacks the DNA-binding domain of E. coli PutA (EcPutA). Consistent with this prediction, purified BjPutA does not exhibit DNA-binding activity in native gel mobility shift assays with promoter regions of the putA gene from B. japonicum. The catalytic and redox properties of BjPutA were characterized and a reduction potential (E(m)) value of -0.132 V (pH 7.5) was determined for the bound FAD/FADH(2) couple in BjPutA that is significantly more negative ( approximately 55 mV) than the E(m) for EcPutA-bound FAD. The more negative E(m) value thermodynamically limits proline reduction of the FAD cofactor in BjPutA. In the presence of phospholipids, reduction of BjPutA is stimulated, suggesting lipids influence the FAD redox environment. Accordingly, an E(m) value of -0.114 V (pH 7.5) was determined for BjPutA-bound FAD in the presence of polar lipids. The molecular basis for the lower reduction potential of FAD in BjPutA relative to EcPutA was explored by site-directed mutagenesis. Amino acid sequence alignment between BjPutA and EcPutA indicates only one difference in active site residues near the isoalloxazine ring of FAD: Val402 in EcPutA is substituted at the analogous position in BjPutA with Ala310. Replacement of A310 by Val in the BjPutA mutant A310V raised the reduction potential of bound FAD relative to wild-type BjPutA to an E(m) value of -0.09 V (pH 7.5). The >40-mV positive shift in the potential of the BjPutA mutant A310V suggests that the corresponding Val residue in Ec

  19. Carbaryl residues in maize products

    International Nuclear Information System (INIS)

    Zayed, S.M.A.D.; Mansour, S.A.; Mostafa, I.Y.; Hassan, A.

    1976-01-01

    The 14 C-labelled insecticide carbaryl was synthesized from [1- 14 C]-1-naphthol at a specific activity of 3.18mCig -1 . Maize plants were treated with the labelled insecticide under simulated conditions of agricultural practice. Mature plants were harvested and studied for distribution of total residues in untreated grains as popularly roasted and consumed, and in the corn oil and corn germ products. Total residues found under these conditions in the respective products were 0.2, 0.1, 0.45 and 0.16ppm. (author)

  20. Study of the surface-enhanced Raman spectroscopy of residual impurities in hydroxylamine-reduced silver colloid and the effects of anions on the colloid activity.

    Science.gov (United States)

    Dong, Xiao; Gu, Huaimin; Liu, Fangfang

    2012-03-01

    The paper investigated the residual ions in hydroxylamine-reduced silver colloid (HRSC) and the relationship between the condition of HRSC and the enhanced mechanisms of this colloid. We also detected the SERS of MB and studied the effects of anions on the Raman signal. In the case of HRSC, the bands of residual ions diminish while the bands of Ag-anions increase gradually with increasing the concentrations of Cl(-) and NO(3)(-). It means the affinity of residual ions on the silver surface is weaker than that of Cl(-) and NO(3)(-) and the residual ions are replaced gradually by the added Cl(-) or NO(3)(-). The Raman signal of residual ions can be detected by treatment with anions that do not bind strongly to the silver surface, such as SO(4)(2-). The most intense band of Ag-anions bonds can be also observed when adding weakly binding anions to the colloid. However, the anions which make up the Ag-anions bonds are residual Cl(-) and the effect of weakly binding anions is only to aggregate the silver particles. Residual Cl(-) can be replaced by I(-) which has the highest affinity. From the detection of methylene blue (MB), the effects of anions on the enhancement of Raman signal are discussed in detail, and these findings could make the conditions suitable for detecting analytes in high efficiency. This study will have a profound implication to SERS users about their interpretation of SERS spectra when obtaining these anomalous bands. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. FAKTOR PSIKOLOGIS, LINGKUNGAN DAN BAURAN PEMASARAN YANG MEMPENGARUHI PEMBELIAN SEPEDA MOTOR Studi Pada Konsumen Sepeda Motor Bebek 4-tak Merk Suzuki dan Honda di Kota Banjarmasin

    Directory of Open Access Journals (Sweden)

    Taufiq Adi Rahmanie

    2012-05-01

    Full Text Available ABSTRAK Penelitian ini dilakukan untuk mengetahui faktor psikologis, lingkungan dan bauran pemasaran yang mempengaruhi pembelian sepeda motor bebek 4-tak merk Suzuki dan Honda di Kota Banjarmasin, dan untuk mengetahui variabel yang dominan terhadap keputusan pembelian. Penelitian ini dilakukan di Kota Banjarmasin. Pengambilan sampel dilakukan dengan teknik non random sampling dengan motode convenience sampling, dengan jumlah sampel 706 responden (355 konsumen Suzuki dan  351 konsumen Honda. Tingkat signifikansi yang ditetapkan dalam penelitian ini adalah 5%. Sebanyak 49 item dalam daftar pertanyaan yang diuraikan dalam 12 variabel dalam faktor psikologis, lingkungan dan bauran pemasaran. Pengujian terhadap semua instrumen penelitian menunjukkan tingkat reliabilitas dan validitas yang tinggi. Hasil penelitian ini semua variabel bebas yaitu persepsi (X1, sikap (X2, gaya hidup (X3, kepribadian (X4, budaya (X5, kelas sosial (X6, kelompok referensi (X7, situasi penentu (X8, produk (X9, harga (X10, promosi (X11 dan distribusi (X12 mempunyai korelasi yang signifikan terhadap keputusan pembelian sepeda motor bebek 4-tak merk Suzuki dan Honda. Diantara keduabelas variabel tersebut, variabel persepsi (X1, sikap (X2, gaya hidup (X3, kelompok referensi (X7, situasi penentu (X8 dan produk (X9 berpengaruh secara nyata terhadap keputusan pembelian sepeda motor bebek 4-tak merk Suzuki dan Honda di Kota Banjarmasin. Koefisien determinasi (R2 sebesar 0.591 menunjukkan kontribusi variabel-variabel independen untuk menjelaskan variabel dependen sebesar 59.1%. Sedangkan variabel yang paling dominan pengaruhnya adalah sikap sebesar 39.6%.   Kata kunci: pemasaran, bauran, psikologis ABSTRACT This research has been done to know psychological, environmental, marketing mix to influence 4 stroke bebek motorcycle (Honda and Suzuki buying in Banjarmasin, and also to know the dominant variable on buying decision. This research has been done in Banjarmasin. Writer uses non

  2. Blocking RhoA/ROCK inhibits the pathogenesis of pemphigus vulgaris by suppressing oxidative stress and apoptosis through TAK1/NOD2-mediated NF-κB pathway.

    Science.gov (United States)

    Liang, Junqin; Zeng, Xuewen; Halifu, Yilinuer; Chen, Wenjing; Hu, Fengxia; Wang, Peng; Zhang, Huan; Kang, Xiaojing

    2017-12-01

    Oxidative stress and apoptosis play critical roles in pemphigus vulgaris (PV). The main aim of the present study was to investigate the effects of RhoA/ROCK signaling on UVB-induced oxidative damage, and to delineate the molecular mechanisms involved in the UVB-mediated inflammatory and apoptotic response. In HaCaT cells, we observed that blockage of RhoA/ROCK signaling with the inhibitor CT04 or Y27632 greatly inhibited the UVB-mediated increase in intracellular reactive oxygen species (ROS). Additionally, inhibition of RhoA/ROCK signaling reduced UVB-induced apoptosis, as exemplified by a reduction in DNA fragmentation, and also elevated anti-apoptotic Bcl-2 protein, concomitant with reduced levels of pro-apoptotic protein Bax, caspase-3 cleavage and decreased PARP-1 protein. The release of inflammatory mediators TNF-α, IL-1β, and IL-6 was also attenuated. Mechanically, we observed that blockage of RhoA/ROCK repressed the TAK1/NOD2-mediated NF-κB pathway in HaCaT cells exposed to UVB. Taken together, these data reveal that RhoA/ROCK signaling is one of the regulators contributing to oxidative damage and apoptosis in human keratinocytes, suggesting that RhoA/ROCK signaling has strong potential to be used as a useful therapeutic target in skin diseases including PV.

  3. Additional Burden of Diseases Associated with Cadmium Exposure: A Case Study of Cadmium Contaminated Rice Fields in Mae Sot District, Tak Province, Thailand

    Directory of Open Access Journals (Sweden)

    Nisarat Songprasert

    2015-08-01

    Full Text Available The cadmium (Cd contaminated rice fields in Mae Sot District, Tak Province, Thailand has been one of the major environmental problems in Thailand for the last 10 years. We used disability adjusted life years (DALYs to estimate the burden of disease attributable to Cd in terms of additional DALYs of Mae Sot residents. Cd exposure data included Cd and β2–microglobulin (β2-MG in urine (as an internal exposure dose and estimated cadmium daily intake (as an external exposure dose. Compared to the general Thai population, Mae Sot residents gained 10%–86% DALYs from nephrosis/nephritis, heart diseases, osteoporosis and cancer depending on their Cd exposure type and exposure level. The results for urinary Cd and dietary Cd intake varied according to the studies used for risk estimation. The ceiling effect was observed in results using dietary Cd intake because of the high Cd content in rice grown in the Mae Sot area. The results from β2-MG were more robust with additional DALYs ranging from 36%–86% for heart failure, cerebral infraction, and nephrosis/nephritis. Additional DALYs is a useful approach for assessing the magnitude of environmental Cd exposure. The Mae Sot population lost more healthy life compared to populations living in a non- or less Cd polluted area. This method should be applicable to various types of environmental contamination problems if exposure assessment information is available.

  4. A field-scale study of cadmium phytoremediation in a contaminated agricultural soil at Mae Sot District, Tak Province, Thailand: (1) Determination of Cd-hyperaccumulating plants.

    Science.gov (United States)

    Khaokaew, Saengdao; Landrot, Gautier

    2015-11-01

    The cadmium (Cd) phytoremediation capabilities of Gynura pseudochina, Chromolaena odorata, Conyza sumatrensis, Crassocephalum crepidioides and Nicotiana tabacum were determined by conducting in-situ experiments in a highly Cd-contaminated agricultural field at Mae Sot District, Tak Province, Thailand. Most of these five plant species, which are commonly found in Thailand, previously demonstrated Cd-hyperaccumulating capacities under greenhouse conditions. This study represented an important initial step in determining if any of these plants could, under field-conditions, effectively remove Cd from the Mae Sot contaminated fields, which represent a health threat to thousands of local villagers. All plant species had at least a 95% survival rate on the final harvest day. Additionally, all plant species, except C. odorata, could hyperaccumulate the extractable Cd amounts present in the soil, based on their associated Bioaccumulation Factor (BAF), Translocation Factor (TF), and background Vegetation Factor (VF). Therefore, the four Cd-hyperaccumulating plant species identified in this study may successfully treat a majority of contaminated fields at Mae Sot, as it was previously reported that Cd amounts present in a number of these soils were mostly available. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Additional Burden of Diseases Associated with Cadmium Exposure: A Case Study of Cadmium Contaminated Rice Fields in Mae Sot District, Tak Province, Thailand

    Science.gov (United States)

    Songprasert, Nisarat; Sukaew, Thitiporn; Kusreesakul, Khanitta; Swaddiwudhipong, Witaya; Padungtod, Chantana; Bundhamcharoen, Kanitta

    2015-01-01

    The cadmium (Cd) contaminated rice fields in Mae Sot District, Tak Province, Thailand has been one of the major environmental problems in Thailand for the last 10 years. We used disability adjusted life years (DALYs) to estimate the burden of disease attributable to Cd in terms of additional DALYs of Mae Sot residents. Cd exposure data included Cd and β2–microglobulin (β2-MG) in urine (as an internal exposure dose) and estimated cadmium daily intake (as an external exposure dose). Compared to the general Thai population, Mae Sot residents gained 10%–86% DALYs from nephrosis/nephritis, heart diseases, osteoporosis and cancer depending on their Cd exposure type and exposure level. The results for urinary Cd and dietary Cd intake varied according to the studies used for risk estimation. The ceiling effect was observed in results using dietary Cd intake because of the high Cd content in rice grown in the Mae Sot area. The results from β2-MG were more robust with additional DALYs ranging from 36%–86% for heart failure, cerebral infarction, and nephrosis/nephritis. Additional DALYs is a useful approach for assessing the magnitude of environmental Cd exposure. The Mae Sot population lost more healthy life compared to populations living in a non- or less Cd polluted area. This method should be applicable to various types of environmental contamination problems if exposure assessment information is available. PMID:26262629

  6. Residual gas analysis

    International Nuclear Information System (INIS)

    Berecz, I.

    1982-01-01

    Determination of the residual gas composition in vacuum systems by a special mass spectrometric method was presented. The quadrupole mass spectrometer (QMS) and its application in thin film technology was discussed. Results, partial pressure versus time curves as well as the line spectra of the residual gases in case of the vaporization of a Ti-Pd-Au alloy were demonstrated together with the possible construction schemes of QMS residual gas analysers. (Sz.J.)

  7. Earthworm tolerance to residual agricultural pesticide contamination

    DEFF Research Database (Denmark)

    Givaudan, Nicolas; Binet, Françoise; Le Bot, Barbara

    2014-01-01

    This study investigates if acclimatization to residual pesticide contamination in agricultural soils is reflected in detoxification, antioxidant enzyme activities and energy budget of earthworms. Five fields within a joint agricultural area exhibited different chemical and farming histories from...

  8. Residue Phe112 of the Human-Type Corrinoid Adenosyltransferase (PduO) Enzyme of Lactobacillus reuteri Is Critical to the Formation of the Four-Coordinate Co(II) Corrinoid Substrate and to the Activity of the Enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Mera, Paola E.; St. Maurice, Martin; Rayment, Ivan; Escalante-Semerena, Jorge C.; UW

    2009-06-08

    ATP:Corrinoid adenosyltransferases (ACAs) catalyze the transfer of the adenosyl moiety from ATP to cob(I)alamin via a four-coordinate cob(II)alamin intermediate. At present, it is unknown how ACAs promote the formation of the four-coordinate corrinoid species needed for activity. The published high-resolution crystal structure of the ACA from Lactobacillus reuteri (LrPduO) in complex with ATP and cob(II)alamin shows that the environment around the alpha face of the corrin ring consists of bulky hydrophobic residues. To understand how these residues promote the generation of the four-coordinate cob(II)alamin, variants of the human-type ACA enzyme from L. reuteri (LrPduO) were kinetically and structurally characterized. These studies revealed that residue Phe112 is critical in the displacement of 5,6-dimethylbenzimidazole (DMB) from its coordination bond with the Co ion of the ring, resulting in the formation of the four-coordinate species. An F112A substitution resulted in a 80% drop in the catalytic efficiency of the enzyme. The explanation for this loss of activity was obtained from the crystal structure of the mutant protein, which showed cob(II)alamin bound in the active site with DMB coordinated to the cobalt ion. The crystal structure of an LrPduO(F112H) variant showed a DMB-off/His-on interaction between the corrinoid and the enzyme, whose catalytic efficiency was 4 orders of magnitude lower than that of the wild-type protein. The analysis of the kinetic parameters of LrPduO(F112H) suggests that the F112H substitution negatively impacts product release. Substitutions of other hydrophobic residues in the Cbl binding pocket did not result in significant defects in catalytic efficiency in vitro; however, none of the variant enzymes analyzed in this work supported AdoCbl biosynthesis in vivo.

  9. Agricultural pesticide residues

    International Nuclear Information System (INIS)

    Fuehr, F.

    1984-01-01

    The utilization of tracer techniques in the study of agricultural pesticide residues is reviewed under the following headings: lysimeter experiments, micro-ecosystems, translocation in soil, degradation of pesticides in soil, biological availability of soil-applied substances, bound residues in the soil, use of macro- and microautography, double and triple labelling, use of tracer labelling in animal experiments. (U.K.)

  10. Inhibition of Glycogen Synthase Kinase-3ß Enhances Cognitive Recovery after Stroke: The Role of TAK1

    Science.gov (United States)

    Venna, Venugopal Reddy; Benashski, Sharon E.; Chauhan, Anjali; McCullough, Louise D.

    2015-01-01

    Memory deficits are common among stroke survivors. Identifying neuroprotective agents that can prevent memory impairment or improve memory recovery is a vital area of research. Glycogen synthase kinase-3ß (GSK-3ß) is involved in several essential intracellular signaling pathways. Unlike many other kinases, GSK-3ß is active only when…

  11. Molecular dissection of the C-terminal regulatory domain of the plant plasma membrane H+-ATPase AHA2: Mapping of residues that when altered give rise to an activated enzyme

    DEFF Research Database (Denmark)

    Axelsen, K.B.; Venema, K.; Jah, T.

    1999-01-01

    The plasma membrane H+-ATPase is a proton pump belonging to the P-type ATPase superfamily and is important for nutrient acquisition in plants, The H+-ATPase is controlled by an autoinhibitory C-terminal regulatory domain and is activated by 14-3-3 proteins which bind to this part of the enzyme...... that when altered lead to increased pump activity group together in two regions of the C-terminus, One region stretches from K863 to L885 and includes two residues (Q879 and R880) that are conserved between plant and fungal H+-ATPases. The other region, incorporating S904 to L919, is situated...... in an extension of the C-terminus unique to plant H+-ATPases, Alteration of residues in both regions led to increased binding of yeast 14-3-3 protein to the plasma membrane of transformed cells. Taken together, our data suggest that modification of residues in two regions of the C-terminal regulatory domain...

  12. Multi-decadal shoreline changes on Takú Atoll, Papua New Guinea: Observational evidence of early reef island recovery after the impact of storm waves

    Science.gov (United States)

    Mann, Thomas; Westphal, Hildegard

    2016-03-01

    Hurricanes, tropical cyclones and other high-magnitude events are important steering mechanisms in the geomorphic development of coral reef islands. Sandy reef islands located outside the storm belts are strongly sensitive to the impact of occasional high-magnitude events and show abrupt, commonly erosive geomorphic change in response to such events. Based on the interpretation of remote sensing data, it is well known that the process of landform recovery might take several decades or even longer. However, despite the increasing amount of scientific attention towards short- and long-term island dynamics, the lack of data and models often prevent a robust analysis of the timing and nature of recovery initiation. Here we show how natural island recovery starts immediately after the impact of a high-magnitude event. We analyze multi-temporal shoreline changes on Takú Atoll, Papua New Guinea and combine our findings with a unique set of published field observations (Smithers and Hoeke, 2014). Trends of shoreline change since 1943 and changes in planform island area indicate a long-term accretionary mode for most islands. Apparent shoreline instability is detected for the last decade of analysis, however this can be explained by the impact of storm waves in December 2008 that (temporarily?) masked the long-term trend. The transition from negative to positive rates of change in the aftermath of this storm event is indicative of inherent negative feedback processes that counteract short-term changes in energy input and represent the initiation of island recovery. Collectively, our results support the concept of dynamic rather than static reef islands and clearly demonstrate how short-term processes can influence interpretations of medium-term change.

  13. Poor Linkage to Care Despite Significant Improvement in Access to Early cART in Central Poland - Data from Test and Keep in Care (TAK Project.

    Directory of Open Access Journals (Sweden)

    Justyna D Kowalska

    Full Text Available The main objective of the TAK project is investigating barriers in accessing HIV care after HIV-diagnosis at the CBVCTs of central Poland. Here we describe factors associated with and changes over time in linkage to care and access to cART.Data collected in 2010-2013 in CBVCTs were linked with HIV clinics records using unique identifiers. Individuals were followed from the day of CBVCTs visit until first clinical visit or 4/06/2014. Cox-proportional hazard models were used to identify factors associated with being linked to care and starting cART.In total 232 persons were diagnosed HIV-positive and 144 (62.1% 95%CI: 55.5-68.3 persons were linked to care. There was no change over time in linkage to care (p = 0.48, while time to starting cART decreased (p = 0.02. Multivariate factors associated with a lower rate of linkage to care were hetero/bisexual sexual orientation, lower education, not having an HIV-positive partner and not using condoms in a stable relationship. Multivariate factors associated with starting cART were lower education, recent year of linked to care, and first HIV RNA and CD4 cell count.Benefits of linkage to care, measured by access to early treatment, steadily improved in recent years. However at least 1 in 3 persons aware of their HIV status in central Poland remained outside professional healthcare. Persons at higher risk of remaining outside care, thus target population for future interventions, are bi/heterosexuals and those with lower levels of education.

  14. Anti Myelin Oligodendrocyte Glycoprotein associated Immunoglobulin G (AntiMOG-IgG)-associated Neuromyelitis Optica Spectrum Disorder with Persistent Disease Activity and Residual Cognitive Impairment.

    Science.gov (United States)

    Pandit, Lekha; Nakashima, Ichiro; Mustafa, Sharik; Takahashi, Toshiyuki; Kaneko, Kimhiko

    2017-01-01

    Antibodies targeting myelin oligodendrocyte glycoprotein (MOG) have been recently reported in association with idiopathic inflammatory central nervous system disorders. Initially believed to be a benign disorder, anti MOG-IgG was noted to cause steroid responsive recurrent optic neuritis and isolated longitudinally extensive myelitis. However, there is growing evidence that the disease may be predominantly relapsing, often producing severe visual loss and involving regions other than the spinal cord and optic nerve. We report an adolescent male with an aggressive disease course previously undescribed in anti MOG-IgG-associated disease that left him with residual cognitive dysfunction.

  15. [Residual neuromuscular blockade].

    Science.gov (United States)

    Fuchs-Buder, T; Schmartz, D

    2017-06-01

    Even small degrees of residual neuromuscular blockade, i. e. a train-of-four (TOF) ratio >0.6, may lead to clinically relevant consequences for the patient. Especially upper airway integrity and the ability to swallow may still be markedly impaired. Moreover, increasing evidence suggests that residual neuromuscular blockade may affect postoperative outcome of patients. The incidence of these small degrees of residual blockade is relatively high and may persist for more than 90 min after a single intubating dose of an intermediately acting neuromuscular blocking agent, such as rocuronium and atracurium. Both neuromuscular monitoring and pharmacological reversal are key elements for the prevention of postoperative residual blockade.

  16. TENORM: Wastewater Treatment Residuals

    Science.gov (United States)

    Water and wastes which have been discharged into municipal sewers are treated at wastewater treatment plants. These may contain trace amounts of both man-made and naturally occurring radionuclides which can accumulate in the treatment plant and residuals.

  17. Comparison of Postural Stubility and Auditory Short- Term Memory Taks Interference between Patients with Functional Ankle Instability and Healthy Individuals

    Directory of Open Access Journals (Sweden)

    Maryam Hayati

    2010-04-01

    Full Text Available Objective: With increased recognition of the importance of conscious process and cognition activities in the stability and regulation of posture and also in line of probable adaptation in central nervous system in patients with functional ankle instability, this study was aimed to assess interference patterns between short–term memory task and postural stability tasks in functional ankle instability patients in comparison with healthy individuals. Materials & Methods: In this cross–sectional and case–control study, with dual–task paradigm fifteen functional ankle instability patients whom were selected simply and conveniently and fifteen healthy persons whom were matched with patients as control group were assessed and compared. Six steps of tests were performed for every case with two situation of balance Biodex system instability and three situation of auditory short–term memory task. Data were analyzed by repeated measurement ANOVA. Results: Finding showed that there was significant interference between postural task and cognitive task in both groups (P<0.05 , but there was no significant difference between interferences of two groups. Conclusion: Present study, certificated disruption in information processing in a peripheral musculoskeletal injury such as functional ankle instability. Application of training programs in order to facilitate central information processing in balance rehabilitation process of this patients and improved dual–task paradigms with different degrees of cognitive and postural tasks seems to be effective.

  18. Residuation in orthomodular lattices

    Directory of Open Access Journals (Sweden)

    Chajda Ivan

    2017-04-01

    Full Text Available We show that every idempotent weakly divisible residuated lattice satisfying the double negation law can be transformed into an orthomodular lattice. The converse holds if adjointness is replaced by conditional adjointness. Moreover, we show that every positive right residuated lattice satisfying the double negation law and two further simple identities can be converted into an orthomodular lattice. In this case, also the converse statement is true and the corresponence is nearly one-to-one.

  19. Characterization of Hospital Residuals

    International Nuclear Information System (INIS)

    Blanco Meza, A.; Bonilla Jimenez, S.

    1997-01-01

    The main objective of this investigation is the characterization of the solid residuals. A description of the handling of the liquid and gassy waste generated in hospitals is also given, identifying the source where they originate. To achieve the proposed objective the work was divided in three stages: The first one was the planning and the coordination with each hospital center, in this way, to determine the schedule of gathering of the waste can be possible. In the second stage a fieldwork was made; it consisted in gathering the quantitative and qualitative information of the general state of the handling of residuals. In the third and last stage, the information previously obtained was organized to express the results as the production rate per day by bed, generation of solid residuals for sampled services, type of solid residuals and density of the same ones. With the obtained results, approaches are settled down to either determine design parameters for final disposition whether for incineration, trituration, sanitary filler or recycling of some materials, and storage politics of the solid residuals that allow to determine the gathering frequency. The study concludes that it is necessary to improve the conditions of the residuals handling in some aspects, to provide the cleaning personnel of the equipment for gathering disposition and of security, minimum to carry out this work efficiently, and to maintain a control of all the dangerous waste, like sharp or polluted materials. In this way, an appreciable reduction is guaranteed in the impact on the atmosphere. (Author) [es

  20. Mutational properties of amino acid residues: implications for evolvability of phosphorylatable residues

    DEFF Research Database (Denmark)

    Creixell, Pau; Schoof, Erwin M.; Tan, Chris Soon Heng

    2012-01-01

    in terms of their mutational activity. Moreover, we highlight the importance of the genetic code and physico-chemical properties of the amino acid residues as likely causes of these inequalities and uncover serine as a mutational hot spot. Finally, we explore the consequences that these different......; it is typically assumed that all amino acid residues are equally likely to mutate or to result from a mutation. Here, by reconstructing ancestral sequences and computing mutational probabilities for all the amino acid residues, we refute this assumption and show extensive inequalities between different residues...... mutational properties have on phosphorylation site evolution, showing that a higher degree of evolvability exists for phosphorylated threonine and, to a lesser extent, serine in comparison with tyrosine residues. As exemplified by the suppression of serine's mutational activity in phosphorylation sites, our...

  1. Interaction of acid mine drainage with Ordinary Portland Cement blended solid residues generated from active treatment of acid mine drainage with coal fly ash.

    Science.gov (United States)

    Gitari, Wilson M; Petrik, Leslie F; Key, David L; Okujeni, Charles

    2011-01-01

    Fly ash (FA) has been investigated as a possible treatment agent for Acid mine drainage (AMD) and established to be an alternative, cheap and economically viable agent compared to the conventional alkaline agents. However, this treatment option also leads to generation of solid residues (SR) that require disposal and one of the proposed disposal method is a backfill in coal mine voids. In this study, the interaction of the SR with AMD that is likely to be present in such backfill scenario was simulated by draining columns packed with SR and SR + 6% Ordinary Portland Cement (OPC) unsaturated with simulated AMD over a 6 month period. The evolving geochemistry of the liquid/solid (L/S) system was evaluated in-terms of the mineral phases likely or controlling contaminants attenuation at the different pH regimes generated. Stepwise acidification of the percolates was observed as the drainage progressed. Two pH buffer zones were observed (7.5-9 and 3-4) for SR and (11.2-11.3 and 3.5-4) for SR + 6% OPC. The solid residue cores (SR) appeared to have a significant buffering capacity, maintaining a neutral to slightly alkaline pH in the leachates for an extended period of time (97 days: L/S 4.3) while SR + 6% OPC reduced this neutralization capacity to 22 days (L/S 1.9). Interaction of AMD with SR or SR + 6% OPC generated alkaline conditions that favored precipitation of Fe, Al, Mn-(oxy) hydroxides, Fe and Ca-Al hydroxysulphates that greatly contributed to the contaminants removal. However, precipitation of these phases was restricted to the pH of the leachates remaining at neutral to circum-neutral levels. Backfill of mine voids with SR promises to be a feasible technology for the disposal of the SR but its success will greatly depend on the disposal scenario, AMD generated and the alkalinity generating potential of the SR. A disadvantage would be the possible re-dissolution of the precipitated phases at pH water column. However extrapolation of this concept to a field

  2. Dominant negative mutant of ionotropic glutamate receptor subunit GluR3: implications for the role of a cysteine residue for its channel activity and pharmacological properties.

    Science.gov (United States)

    Watase, K; Sekiguchi, M; Matsui, T A; Tagawa, Y; Wada, K

    1997-01-01

    We reported that a 33-amino-acid deletion (from tyrosine-715 to glycine-747) in a putative extracellular loop of GluR3 produced a mutant that exhibited dominant negative effects upon the functional expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors [Sekiguchi et al. (1994) J. Biol. Chem. 269, 14559-14565]. In this study, we searched for a key residue in the dominant negative effects to explore the mechanism and examined the role of the residue in the function of the AMPA receptor. We prepared 20 GluR3 mutants with amino acid substitutions within the 33-amino-acid-region, and dominant negative effects were tested electrophysiologically in Xenopus oocytes co-expressing the mutant and normal subunits. Among the mutants, only a GluR3 mutant in which an original cysteine (Cys)-722 was replaced by alanine exhibited a dominant negative effect comparable with that of the original mutant in which the entire 33-amino-acid segment is deleted. The co-expression of the Cys-722 mutant did not inhibit the translation of normal subunits in oocytes. The Cys-722 mutant formed a functional homomeric receptor with significantly higher affinity for glutamate or kainate than a homomeric GluR3 receptor. The Cys-722 mutation greatly enhanced the sensitivity of GluR3 for aniracetam, which alters kinetic properties of AMPA receptors. The kainate-induced currents in oocytes expressing the Cys-722 mutant alone showed strong inward rectification. These results suggest that the Cys-722 in GluR3 is important for dominant negative effects and plays a crucial role in the determination of pharmacological properties in AMPA receptor function. PMID:9065754

  3. Calcination/dissolution residue treatment

    International Nuclear Information System (INIS)

    Knight, R.C.; Creed, R.F.; Patello, G.K.; Hollenberg, G.W.; Buehler, M.F.; O'Rourke, S.M.; Visnapuu, A.; McLaughlin, D.F.

    1994-09-01

    Currently, high-level wastes are stored underground in steel-lined tanks at the Hanford site. Current plans call for the chemical pretreatment of these wastes before their immobilization in stable glass waste forms. One candidate pretreatment approach, calcination/dissolution, performs an alkaline fusion of the waste and creates a high-level/low-level partition based on the aqueous solubilities of the components of the product calcine. Literature and laboratory studies were conducted with the goal of finding a residue treatment technology that would decrease the quantity of high-level waste glass required following calcination/dissolution waste processing. Four elements, Fe, Ni, Bi, and U, postulated to be present in the high-level residue fraction were identified as being key to the quantity of high-level glass formed. Laboratory tests of the candidate technologies with simulant high-level residues showed reductive roasting followed by carbonyl volatilization to be successful in removing Fe, Ni, and Bi. Subsequent bench-scale tests on residues from calcination/dissolution processing of genuine Hanford Site tank waste showed Fe was separated with radioelement decontamination factors of 70 to 1,000 times with respect to total alpha activity. Thermodynamic analyses of the calcination of five typical Hanford Site tank waste compositions also were performed. The analyses showed sodium hydroxide to be the sole molten component in the waste calcine and emphasized the requirement for waste blending if fluid calcines are to be achieved. Other calcine phases identified in the thermodynamic analysis indicate the significant thermal reconstitution accomplished in calcination

  4. Extracellular acid block and acid-enhanced inactivation of the Ca2+-activated cation channel TRPM5 involve residues in the S3-S4 and S5-S6 extracellular domains.

    Science.gov (United States)

    Liu, Dan; Zhang, Zheng; Liman, Emily R

    2005-05-27

    TRPM5, a member of the superfamily of transient receptor potential ion channels, is essential for the detection of bitter, sweet, and amino acid tastes. In heterologous cell types it forms a nonselective cation channel that is activated by intracellular Ca(2+). TRPM5 is likely to be part of the taste transduction cascade, and regulators of TRPM5 are likely to affect taste sensation. In this report we show that TRPM5, but not the related channel TRPM4b, is potently blocked by extracellular acidification. External acidification has two effects, a fast reversible block of the current (IC(50) pH = 6.2) and a slower irreversible enhancement of current inactivation. Mutation of a single Glu residue in the S3-S4 linker and a His residue in the pore region each reduced sensitivity of TRPM5 currents to fast acid block (IC(50) pH = 5.8 for both), and the double mutant was nearly insensitive to acidic pH (IC(50) pH = 5.0). Prolonged exposure to acidic pH enhanced inactivation of TRPM5 currents, and mutant channels that were less sensitive to acid block were also less sensitive to acid-enhanced inactivation, suggesting an intimate association between the two processes. These processes are, however, distinct because the pore mutant H896N, which has normal sensitivity to acid block, shows significant recovery from acid-enhanced inactivation. These data show that extracellular acidification acts through specific residues on TRPM5 to block conduction through two distinct but related mechanisms and suggest a possible interaction between extracellular pH and activation and adaptation of bitter, sweet, and amino acid taste transduction.

  5. Atividade residual de herbicidas pré-emergentes aplicados na cultura da soja sobre o milheto cultivado em sucessão Effect of the residual activity of pre-emergent herbicides applied in soybean on pearl millet cultivated in succession

    Directory of Open Access Journals (Sweden)

    H.A. Dan

    2011-06-01

    after sowing of soybean Msoy-6101, in plots of 80 m2, divided into sub-plots of 20 m2 (5 x 4 m. A randomized block design with four replications was used in a split-plot scheme 5 x 4. In the sub-plots, pearl millet hybrid ADR-7010 was sown in four periods after herbicide application, corresponding to 0, 40, 80 and 120 days after herbicide application (DAA. Pear millet injury, stand, height and dry biomass of the plant were evaluated at 7, 15, and 28 days after emergence; crop grain yield was evaluated at the end of the cycle. Pearl millet hybrid ADR-7010 showed high sensitivity to the residual activity of sulfentrazone, imazaquin, and diclosulam when grown immediately after herbicide application. The bioactivity of imazaquin, diclosulam, and flumioxazin was not sufficient to affect grain yield of millet grown after soybean (120 DAA, suggesting that this interval is sufficiently long for dissipation of these herbicides. Among the pre-emergent herbicides, sulfentrazone showed longer residual activity, negatively influencing the crop's yield during the time interval studied.

  6. Farvel og tak til naturfagene

    DEFF Research Database (Denmark)

    Weirsøe, Mathilde

    2011-01-01

    Lektor Helene Sørensen takker af efter 24 års arbejde for at fremme interessem for natur og teknik, særligt blandt pigerne. Hun går på pension, men det sker med en vis portion vemod, mest fordi »ingen har passion for at gøre undervisningen i naturfag bedre«....

  7. Corn residue removal and CO2 emissions

    Science.gov (United States)

    Carbon dioxide (CO2), nitrous oxide (N2O), and methane (CH4) are the primary greenhouse gases (GHG) emitted from the soil due to agricultural activities. In the short-term, increases in CO2 emissions indicate increased soil microbial activity. Soil micro-organisms decompose crop residues and release...

  8. Residual-stress measurements

    Energy Technology Data Exchange (ETDEWEB)

    Ezeilo, A.N.; Webster, G.A. [Imperial College, London (United Kingdom); Webster, P.J. [Salford Univ. (United Kingdom)

    1997-04-01

    Because neutrons can penetrate distances of up to 50 mm in most engineering materials, this makes them unique for establishing residual-stress distributions non-destructively. D1A is particularly suited for through-surface measurements as it does not suffer from instrumental surface aberrations commonly found on multidetector instruments, while D20 is best for fast internal-strain scanning. Two examples for residual-stress measurements in a shot-peened material, and in a weld are presented to demonstrate the attractive features of both instruments. (author).

  9. 1H, 13C, and 15N resonance assignments of an enzymatically active domain from the catalytic component (CDTa, residues 216-420) of a binary toxin from Clostridium difficile.

    Science.gov (United States)

    Roth, Braden M; Godoy-Ruiz, Raquel; Varney, Kristen M; Rustandi, Richard R; Weber, David J

    2016-04-01

    Clostridium difficile is a bacterial pathogen and is the most commonly reported source of nosocomial infection in industrialized nations. Symptoms of C. difficile infection (CDI) include antibiotic-associated diarrhea, pseudomembranous colitis, sepsis and death. Over the last decade, rates and severity of hospital infections in North America and Europe have increased dramatically and correlate with the emergence of a hypervirulent strain of C. difficile characterized by the presence of a binary toxin, CDT (C. difficile toxin). The binary toxin consists of an enzymatic component (CDTa) and a cellular binding component (CDTb) that together form the active binary toxin complex. CDTa harbors a pair of structurally similar but functionally distinct domains, an N-terminal domain (residues 1-215; (1-215)CDTa) that interacts with CDTb and a C-terminal domain (residues 216-420; (216-420)CDTa) that harbors the intact ADP-ribosyltransferase (ART) active site. Reported here are the (1)H, (13)C, and (15)N backbone resonance assignments of the 23 kDa, 205 amino acid C-terminal enzymatic domain of CDTa, termed (216-420)CDTa. These NMR resonance assignments for (216-420)CDTa represent the first for a family of ART binary toxins and provide the framework for detailed characterization of the solution-state protein structure determination, dynamic studies of this domain, as well as NMR-based drug discovery efforts.

  10. Cyolane residues in milk of lactating goats

    International Nuclear Information System (INIS)

    Zayed, S.M.A.D.; Osman, A.; Fakhr, I.M.I.

    1981-01-01

    Consecutive feeding of lactating goats with 14 C-alkyl labelled cyolane for 5 days at dietary levels 8 and 16 ppm resulted in the appearance of measurable insecticide residues in milk (0.02-0.04 mg/kg). The residue levels were markedly reduced after a withdrawal period of 7 days. Analysis of urine and milk residues showed the presence of similar metabolites in addition to the parent compound. The major part of the residue consisted of mono-, diethyl phosphate and 2 hydrophilic unknown metabolites. The erythrocyte cholinesterase activity was reduced to about 50% after 24 hours whereas the plasma enzyme was only slightly affected. The animals remained symptom-free during the experimental period. (author)

  11. Molecular insights into the differences in anti-inflammatory activities of green tea catechins on IL-1β signaling in rheumatoid arthritis synovial fibroblasts.

    Science.gov (United States)

    Fechtner, Sabrina; Singh, Anil; Chourasia, Mukesh; Ahmed, Salahuddin

    2017-08-15

    In this study, we found that catechins found in green tea (EGCG, EGC, and EC) differentially interfere with the IL-1β signaling pathway which regulates the expression of pro-inflammatory mediators (IL-6 and IL-8) and Cox-2 in primary human rheumatoid arthritis synovial fibroblasts (RASFs). EGCG and EGC inhibited IL-6, IL-8, and MMP-2 production and selectively inhibited Cox-2 expression. EC did not exhibit any inhibitory effects. When we looked at the expression of key signaling proteins in the IL-1β signaling pathway, we found all the tested catechins could inhibit TAK-1 activity. Therefore, the consumption of green tea offers an overall anti-inflammatory effect. Molecular docking analysis confirms that EGCG, EGC, and EC all occupy the active site of the TAK1 kinase domain. However, EGCG occupies the majority of the TAK1 active site. In addition to TAK1 inhibition, EGCG can also inhibit P38 and nuclear NF-κB expression whereas EC and EGC were not effective inhibitors. Our findings suggest one of the main health benefits associated with the consumption of green tea are due to the activity of EGCG and EGC which are both present at higher amounts. Although EGCG is the most effective catechin at inhibiting downstream inflammatory signaling, its effectiveness could be hindered by the presence of EC. Therefore, varying EC content in green tea may reduce the anti-inflammatory effects of other potential catechins in green tea. Copyright © 2017. Published by Elsevier Inc.

  12. Determination of activity by gamma spectrometry of radionuclides present in drums of residues generated in nuclear centrals; Determinacion de actividad por espectrometria gamma de radionucleidos presentes en tambores de residuos generados en centrales nucleares

    Energy Technology Data Exchange (ETDEWEB)

    Aguiar, J.C.; Fernandez, J. [Autoridad Regulatoria Nuclear, Av. Del Libertador 8250, Ciudad Autonoma de Buenos Aires (Argentina)]. e-mail: jaguiar@cae.arn.gov.ar

    2006-07-01

    The generation of radioactive residuals in nuclear centrals as CNA I (Atucha I Nuclear Central) and CNE (Embalse Nuclear Central) makes that the measurement of those radionuclides has been a previous stage to the waste management. A method used in those nuclear centrals it is the gamma spectrometry with HPGe detectors, previous to the immobilization of the residual in a cemented matrix, with this the contact with the external agents and its possible dispersion to the atmosphere in the short term is avoided. The ARN (Nuclear Regulatory Authority) of Argentina it carries out periodically intercomparisons and evaluations of the measurement and procedures systems used in the nuclear power stations for the correct measurement and determination of activity of radioactive residuals by gamma spectrometry. In this work an independent method of measurement is exposed to the nuclear power stations. To determine the activity of the residuals by gamma spectrometry deposited in drums, it is required of the precise knowledge of the efficiency curve for such geometry and matrix. Due to the RNA doesn't have a pattern of these characteristics, a mathematical model has been used to obtain this efficiency curve. For it, it is necessary to determine previously: 1) the geometric efficiency or solid angle sustained by the source-detector system (drum-detector) applying a mathematical model described in this work. 2) To estimate the auto-attenuation factor that present the photons in the cemented matrix, these calculations are carried out with a simple equation and its are verified with the Micro Shield 6.10 program. The container commonly used by these nuclear power stations its are drums for 220 liters constructed with SAE 1010 steel and with a thickness of 0.127 cm, with an approximate weight 7.73 Kg., internal diameter of 57.1 cm, and height: 87 cm. The results obtained until the moment register a discrepancy from 5 to 10% with relationship to the measurements carried out by the

  13. Methyl bromide residues in fumigated cocoa beans

    International Nuclear Information System (INIS)

    Adomako, D.

    1975-01-01

    The 14 C activity in unroasted [ 14 C]-methyl bromide fumigated cocoa beans was used to study the fate and persistence of CH 3 Br in the stored beans. About 70% of the residues occurred in the shells. Unchanged CH 3 Br could not be detected, all the sorbed CH 3 Br having reacted with bean constituents apparently to form 14 C-methylated derivatives and inorganic bromide. No 14 C activity was found in the lipid fraction. Roasting decreased the bound (non-volatile) residues, with corresponding changes in the activities and amounts of free sugars, free and protein amino acids. Roasted nibs and shells showed a two-fold increase in the volatile fraction of the 14 C residue. This fraction may be related to the volatile aroma compounds formed by Maillard-type reactions. (author)

  14. Co-composting of oil exhausted olive-cake, poultry manure and industrial residues of agro-food activity for soil amendment.

    Science.gov (United States)

    Sellami, F; Jarboui, R; Hachicha, S; Medhioub, K; Ammar, E

    2008-03-01

    The co-composting of exhausted olive-cake with poultry manure and sesame shells was investigated. These organic solid wastes were watered by the confectionary wastewater which is characterized by its high content of residual sugars raising its COD. Four aerated windrows were performed to establish the effects of confectionary by-products on the compost process. Different mixtures of the agro-industrial wastes were used. During the composting process, physico-chemical parameters (temperature, moisture, pH, electrical conductivity, total carbon and total nitrogen) were studied. The stability of the biological system was noticed after 70 days. The final products were characterized by their relatively high organic matter content, and low C/N ratio of 14-17. The humidification of the windrows with the wastewater seemed to have accelerated the composting process in comparison to a windrow humidified with water. In addition, the organic matter degradation was enhanced to reach 55-70%. The application of the obtained composts to soil appeared to significantly improve the soil fertility. Indeed, field experiments showed an increase in potato yield; the production was 30.5-37.5 tons ha(-1), compared to 30.5 tons ha(-1) with farm manure.

  15. Activation of Bacillus subtilis Ugd by the BY-Kinase PtkA Proceeds via Phosphorylation of Its Residue Tyrosine 70

    DEFF Research Database (Denmark)

    Petranovic, Dina; Grangeasse, C.; Macek, B.

    2009-01-01

    The phosphorylation-dependent activation of bacterial UDP-glucose dehydrogenases by BY-kinases has been previously described in several bacterial model organisms, but the identity of phosphorylated tyrosine(s) and the exact activation mechanism remained unknown. A recent site-specific phosphoprot......The phosphorylation-dependent activation of bacterial UDP-glucose dehydrogenases by BY-kinases has been previously described in several bacterial model organisms, but the identity of phosphorylated tyrosine(s) and the exact activation mechanism remained unknown. A recent site...

  16. Composition of carbonization residues

    Energy Technology Data Exchange (ETDEWEB)

    Hupfer; Leonhardt

    1943-11-27

    This report compared the composition of samples from Wesseling and Leuna. In each case the sample was a residue from carbonization of the residues from hydrogenation of the brown coal processed at the plant. The composition was given in terms of volatile components, fixed carbon, ash, water, carbon, hydrogen, oxygen, nitrogen, volatile sulfur, and total sulfur. The result of carbonization was given in terms of (ash and) coke, tar, water, gas and losses, and bitumen. The composition of the ash was given in terms of silicon dioxide, ferric oxide, aluminum oxide, calcium oxide, magnesium oxide, potassium and sodium oxides, sulfur trioxide, phosphorus pentoxide, chlorine, and titanium oxide. The most important difference between the properties of the two samples was that the residue from Wesseling only contained 4% oil, whereas that from Leuna had about 26% oil. Taking into account the total amount of residue processed yearly, the report noted that better carbonization at Leuna could save 20,000 metric tons/year of oil. Some other comparisons of data included about 33% volatiles at Leuna vs. about 22% at Wesseling, about 5 1/2% sulfur at Leuna vs. about 6 1/2% at Leuna, but about 57% ash for both. Composition of the ash differed quite a bit between the two. 1 table.

  17. Natural radioactivity in petroleum residues

    International Nuclear Information System (INIS)

    Gazineu, M.H.P.; Gazineu, M.H.P.; Hazin, C.A.; Hazin, C.A.

    2006-01-01

    The oil extraction and production industry generates several types of solid and liquid wastes. Scales, sludge and water are typical residues that can be found in such facilities and that can be contaminated with Naturally Occurring Radioactive Material (N.O.R.M.). As a result of oil processing, the natural radionuclides can be concentrated in such residues, forming the so called Technologically Enhanced Naturally Occurring Radioactive Material, or T.E.N.O.R.M.. Most of the radionuclides that appear in oil and gas streams belong to the 238 U and 232 Th natural series, besides 40 K. The present work was developed to determine the radionuclide content of scales and sludge generated during oil extraction and production operations. Emphasis was given to the quantification of 226 Ra, 228 Ra and 40 K since these radionuclides,are responsible for most of the external exposure in such facilities. Samples were taken from the P.E.T.R.O.B.R.A.S. unity in the State of Sergipe, in Northeastern Brazil. They were collected directly from the inner surface of water pipes and storage tanks, or from barrels stored in the waste storage area of the E and P unit. The activity concentrations for 226 Ra, 228 Ra and 40 K were determined by using an HP Ge gamma spectrometric system. The results showed concentrations ranging from 42.7 to 2,110.0 kBq/kg for 226 Ra, 40.5 to 1,550.0 kBq/kg for 228 Ra, and 20.6 to 186.6 kBq/kg for 40 K. The results highlight the importance of determining the activity concentration of those radionuclides in oil residues before deciding whether they should be stored or discarded to the environment. (authors)

  18. Natural radioactivity in petroleum residues

    Energy Technology Data Exchange (ETDEWEB)

    Gazineu, M.H.P. [UNICAP, Dept. de Quimica, Recife (Brazil); Gazineu, M.H.P.; Hazin, C.A. [UFPE, Dept. de Energia Nuclear, Recife (Brazil); Hazin, C.A. [Centro Regional de Ciencias Nucleares/ CNEN, Recife (Brazil)

    2006-07-01

    The oil extraction and production industry generates several types of solid and liquid wastes. Scales, sludge and water are typical residues that can be found in such facilities and that can be contaminated with Naturally Occurring Radioactive Material (N.O.R.M.). As a result of oil processing, the natural radionuclides can be concentrated in such residues, forming the so called Technologically Enhanced Naturally Occurring Radioactive Material, or T.E.N.O.R.M.. Most of the radionuclides that appear in oil and gas streams belong to the {sup 238}U and {sup 232}Th natural series, besides 40 K. The present work was developed to determine the radionuclide content of scales and sludge generated during oil extraction and production operations. Emphasis was given to the quantification of {sup 226}Ra, {sup 228}Ra and 40 K since these radionuclides,are responsible for most of the external exposure in such facilities. Samples were taken from the P.E.T.R.O.B.R.A.S. unity in the State of Sergipe, in Northeastern Brazil. They were collected directly from the inner surface of water pipes and storage tanks, or from barrels stored in the waste storage area of the E and P unit. The activity concentrations for {sup 226}Ra, {sup 228}Ra and 40 K were determined by using an HP Ge gamma spectrometric system. The results showed concentrations ranging from 42.7 to 2,110.0 kBq/kg for {sup 226}Ra, 40.5 to 1,550.0 kBq/kg for {sup 228}Ra, and 20.6 to 186.6 kBq/kg for 40 K. The results highlight the importance of determining the activity concentration of those radionuclides in oil residues before deciding whether they should be stored or discarded to the environment. (authors)

  19. Residual radioactivity of treated green diamonds.

    Science.gov (United States)

    Cassette, Philippe; Notari, Franck; Lépy, Marie-Christine; Caplan, Candice; Pierre, Sylvie; Hainschwang, Thomas; Fritsch, Emmanuel

    2017-08-01

    Treated green diamonds can show residual radioactivity, generally due to immersion in radium salts. We report various activity measurements on two radioactive diamonds. The activity was characterized by alpha and gamma ray spectrometry, and the radon emanation was measured by alpha counting of a frozen source. Even when no residual radium contamination can be identified, measurable alpha and high-energy beta emissions could be detected. The potential health impact of radioactive diamonds and their status with regard to the regulatory policy for radioactive products are discussed. Copyright © 2017. Published by Elsevier Ltd.

  20. Contributions of the substrate-binding arginine residues to maleate-induced closure of the active site of Escherichia coli aspartate aminotransferase.

    Science.gov (United States)

    Matharu, A; Hayashi, H; Kagamiyama, H; Maras, B; John, R A

    2001-03-01

    Crystallography shows that aspartate aminotransferase binds dicarboxylate substrate analogues by bonds to Arg292 and Arg386, respectively [Jager, J, Moser, M. Sauder, U. & Jansonius, J. N. (1994) J. Mol. Biol., 239, 285-305]. The contribution of each interaction to the conformational change that the enzyme undergoes when it binds ligands via these residues, is assessed by probing mutant forms of the enzyme lacking either or both arginines. The probes used are NaH(3)BCN which reduces the cofactor imine, the reactive substrate analogue, cysteine sulfinate and proteolysis by trypsin. The unreactive substrate analogue, maleate, is used to induce closure. Each single mutant reacted only 2.5-fold more slowly with NaH(3)BCN than the wild-type indicating that charge repulsion by the arginines contributes little to maintaining the open conformation. Maleate lowered the rate of reduction of the wild-type enzyme more than 300-fold but had little effect on the reaction of the mutant enzymes indicating that the ability of this dicarboxylate analogue to bridge the arginines precisely makes the major contribution to closure. The R292L mutant reacted 20 times more rapidly with cysteine sulfinate than R386L but 5 x 10(4) times more slowly than the wild-type enzyme, consistent with the proposal that enzyme's catalytic abilities are not developed unless closure is induced by bridging of the arginines. Proteolysis of the mutants with trypsin showed that, in the wild-type enzyme, the bonds most susceptible to trypsin are those contributed by Arg292 and Arg386. Proteolysis of the next most susceptible bond, at Arg25 in the double mutant, was protected by maleate demonstrating the presence of an additional site on the enzyme for binding dicarboxylates.

  1. Eviromental Economic and Technological Residues Management Demands: An Optimization Tool.

    Directory of Open Access Journals (Sweden)

    Marisa Soares Borges

    2012-12-01

    Full Text Available Industrial residues management is a very demanding task since many different goals must be achieved. The combination of different approaches used by people from different stuff is very challenging activity that can misuse the residues potential value and applicability. An interactive WEB base tool, to integrate different sectors and overcome residues management difficulties will be presented. The system must be loaded with all data concerning the residue life cycle, and through data integration and modeling routine will give the best alternative as output. As wider and complete the system data becomes, by information loading from differen t segment, more efficient the residues management becomes. The user friendly tool will encourage the participation of industries, labs and research institutions to obtain qualified information about industrial residues inventory, raw materials recovery, characteristics, treatment and alternative uses, to achieve residues management sustainability.

  2. Pesticide residues in locally available cereals and vegetables

    International Nuclear Information System (INIS)

    Cunanan, S.A.; Santos, F.L.; Bonoan, L.S.

    1976-03-01

    Vegetable samples (pechay, cabbage, lettuce, green beans and tomatoes) bought from public markets in the Metro Manila area were analyzed for pesticide residues using gas chromatography. The samples analyzed in 1968-69 contained high levels of chlorinated pesticides such as DDT, Aldrin, Endrin, and Thiodan, while in the samples analyzed in January 1976, no chlorinated and organophosphate pesticides were detected. Cereal samples (rice, corn and sorghum) were obtained from the National Grains Authority and analyzed for pesticide residues and bromine residues. Total bromine residues was determined by neutron activation analysis. In most of the samples analyzed, the concentrations of pesticide residues were below the tolerance levels set by the FAO/WHO Committee on Pesticide Residues in Foods. An exception was one rice sample from Thailand, the bromine residue content (110ppm) of which exceeds the tolerance level of 50ppm

  3. Residual vein thrombosis and onset of post-thrombotic syndrome: influence of the 4G/5G polymorphism of plasminogen activator inhibitor-1 gene.

    Science.gov (United States)

    Incalcaterra, Egle; Meli, Francesco; Muratori, Ida; Corrado, Egle; Amato, Corrado; Canino, Baldassare; Ferrara, Filippo

    2014-03-01

    Plasminogen activator inhibitor-1 (PAI-1) is the most important inhibitor of plasminogen activator. The functional 4G/5G polymorphism of the gene coding for PAI-1 may affect PAI-1 plasmatic activity, influencing the imbalance between coagulation and fibrinolysis cascades. In this prospective cohort analytic study, we investigated the role of this single nucleotide polymorphism in the persistence of thrombotic lesion and the occurrence of post-thrombotic syndrome. In a group of 168 patients with post-surgical deep vein thrombosis of the legs, we analyzed the 4G/5G polymorphism in the promoter of PAI-1 gene and plasmatic PAI-1 activity. Enrolled patients were divided in two groups: patients with 4G/5G polymorphism and increased PAI-1 activity (n=85) and patients without 4G/5G polymorphism and normal PAI-1 activity (n=83). All patients were treated according to current protocols and re-examined after 3, 12 and 36 months in order to evaluate the persistence of thrombotic lesion and the occurrence of post-thrombotic syndrome. We found a significantly increased PAI activity in carrier of the 4G allele, who experienced much more frequently a persistence of thrombosis after 3, 12 and 36 months and/or the development of post-thrombosis syndrome, in spite of the anticoagulant treatment. These data not only confirm the role played by PAI-1 activity and by the 4G/5G SNP of the PAI-1 gene, but also suggest that current therapeutic protocols, recommending the administration of low weight molecular heparin and oral anticoagulant for the treatment of deep vein thrombosis, could be non sufficient for patients genetically predisposed to a less efficient clot lysis. Copyright © 2013. Published by Elsevier Ltd.

  4. Comparative structural modeling of six old yellow enzymes (OYEs from the necrotrophic fungus Ascochyta rabiei: insight into novel OYE classes with differences in cofactor binding, organization of active site residues and stereopreferences.

    Directory of Open Access Journals (Sweden)

    Shadab Nizam

    Full Text Available Old Yellow Enzyme (OYE1 was the first flavin-dependent enzyme identified and characterized in detail by the entire range of physical techniques. Irrespective of this scrutiny, true physiological role of the enzyme remains a mystery. In a recent study, we systematically identified OYE proteins from various fungi and classified them into three classes viz. Class I, II and III. However, there is no information about the structural organization of Class III OYEs, eukaryotic Class II OYEs and Class I OYEs of filamentous fungi. Ascochyta rabiei, a filamentous phytopathogen which causes Ascochyta blight (AB in chickpea possesses six OYEs (ArOYE1-6 belonging to the three OYE classes. Here we carried out comparative homology modeling of six ArOYEs representing all the three classes to get an in depth idea of structural and functional aspects of fungal OYEs. The predicted 3D structures of A. rabiei OYEs were refined and evaluated using various validation tools for their structural integrity. Analysis of FMN binding environment of Class III OYE revealed novel residues involved in interaction. The ligand para-hydroxybenzaldehyde (PHB was docked into the active site of the enzymes and interacting residues were analyzed. We observed a unique active site organization of Class III OYE in comparison to Class I and II OYEs. Subsequently, analysis of stereopreference through structural features of ArOYEs was carried out, suggesting differences in R/S selectivity of these proteins. Therefore, our comparative modeling study provides insights into the FMN binding, active site organization and stereopreference of different classes of ArOYEs and indicates towards functional differences of these enzymes. This study provides the basis for future investigations towards the biochemical and functional characterization of these enigmatic enzymes.

  5. Residual stresses in material processing

    Science.gov (United States)

    Kozaczek, K. J.; Watkins, T. R.; Hubbard, C. R.; Wang, Xun-Li; Spooner, S.

    Material manufacturing processes often introduce residual stresses into the product. The residual stresses affect the properties of the material and often are detrimental. Therefore, the distribution and magnitude of residual stresses in the final product are usually an important factor in manufacturing process optimization or component life prediction. The present paper briefly discusses the causes of residual stresses. It then addresses the direct, nondestructive methods of residual stress measurement by X ray and neutron diffraction. Examples are presented to demonstrate the importance of residual stress measurement in machining and joining operations.

  6. Identification of novel amino acid residues of influenza virus PA-X that are important for PA-X shutoff activity by using yeast.

    Science.gov (United States)

    Oishi, Kohei; Yamayoshi, Seiya; Kawaoka, Yoshihiro

    2018-03-01

    The influenza A virus protein PA-X comprises an N-terminal PA region and a C-terminal PA-X-specific region. PA-X suppresses host gene expression, termed shutoff, via mRNA cleavage. Although the endonuclease active site in the N-terminal PA region of PA-X and basic amino acids in the C-terminal PA-X-specific region are known to be important for PA-X shutoff activity, other amino acids may also play a role. Here, we used yeast to identify novel amino acids of PA-X that are important for PA-X shutoff activity. Unlike wild-type PA-X, most PA-X mutants predominantly localized in the cytoplasm, indicating that these mutations decreased the shutoff activity of PA-X by affecting PA-X translocation to the nucleus. Mapping of the identified amino acids onto the N-terminal structure of PA revealed that some of them likely contribute to the formation of the endonuclease active site of PA. Copyright © 2018. Published by Elsevier Inc.

  7. SRC Residual fuel oils

    Science.gov (United States)

    Tewari, Krishna C.; Foster, Edward P.

    1985-01-01

    Coal solids (SRC) and distillate oils are combined to afford single-phase blends of residual oils which have utility as fuel oils substitutes. The components are combined on the basis of their respective polarities, that is, on the basis of their heteroatom content, to assure complete solubilization of SRC. The resulting composition is a fuel oil blend which retains its stability and homogeneity over the long term.

  8. Composition of carbonization residues

    Energy Technology Data Exchange (ETDEWEB)

    Hupfer; Leonhardt

    1943-11-30

    This report gave a record of the composition of several samples of residues from carbonization of various hydrogenation residue from processing some type of coal or tar in the Bergius process. These included Silesian bituminous coal processed at 600 atm. with iron catalyst, in one case to produce gasoline and middle oil and in another case to produce heavy oil excess, Scholven coal processed at 250 atm. with tin oxalate and chlorine catalyst, Bruex tar processed in a 10-liter oven using iron catalyst, and a pitch mixture from Welheim processed in a 10-liter over using iron catalyst. The values gathered were compared with a few corresponding values estimated for Boehlen tar and Gelsenberg coal based on several assumptions outlined in the report. The data recorded included percentage of ash in the dry residue and percentage of carbon, hydrogen, oxygen, nitrogen, chlorine, total sulfur, and volatile sulfur. The percentage of ash varied from 21.43% in the case of Bruex tar to 53.15% in the case of one of the Silesian coals. Percentage of carbon varied from 44.0% in the case of Scholven coal to 78.03% in the case of Bruex tar. Percentage of total sulfur varied from 2.28% for Bruex tar to a recorded 5.65% for one of the Silesian coals and an estimated 6% for Boehlen tar. 1 table.

  9. The dapE-encoded N-succinyl-L,L-Diaminopimelic Acid Desuccinylase from Haemophilus influenzae Contains two Active Site Histidine Residues

    OpenAIRE

    Gillner, Danuta M.; Bienvenue, David L.; Nocek, Boguslaw P.; Joachimiak, Andrzej; Zachary, Vincentos; Bennett, Brian; Holz, Richard C.

    2008-01-01

    The catalytic and structural properties of the H67A and H349A altered dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE) from H. influenzae were investigated. Based on sequence alignment with CPG2 both H67 and H349 were predicted to be Zn(II) ligands. Catalytic activity was observed for the H67A altered DapE enzyme which exhibited kcat = 1.5 ± 0.5 sec−1 and Km = 1.4 ± 0.3 mM. No catalytic activity was observed for H349A under the experimental conditions used. The EPR and ele...

  10. Sustainable System for Residual Hazards Management

    International Nuclear Information System (INIS)

    Kevin M. Kostelnik; James H. Clarke; Jerry L. Harbour

    2004-01-01

    Hazardous, radioactive and other toxic substances have routinely been generated and subsequently disposed of in the shallow subsurface throughout the world. Many of today's waste management techniques do not eliminate the problem, but rather only concentrate or contain the hazardous contaminants. Residual hazards result from the presence of hazardous and/or contaminated material that remains on-site following active operations or the completion of remedial actions. Residual hazards pose continued risk to humans and the environment and represent a significant and chronic problem that require continuous long-term management (i.e. >1000 years). To protect human health and safeguard the natural environment, a sustainable system is required for the proper management of residual hazards. A sustainable system for the management of residual hazards will require the integration of engineered, institutional and land-use controls to isolate residual contaminants and thus minimize the associated hazards. Engineered controls are physical modifications to the natural setting and ecosystem, including the site, facility, and/or the residual materials themselves, in order to reduce or eliminate the potential for exposure to contaminants of concern (COCs). Institutional controls are processes, instruments, and mechanisms designed to influence human behavior and activity. System failure can involve hazardous material escaping from the confinement because of system degradation (i.e., chronic or acute degradation) or by external intrusion of the biosphere into the contaminated material because of the loss of institutional control. An ongoing analysis of contemporary and historic sites suggests that the significance of the loss of institutional controls is a critical pathway because decisions made during the operations/remedial action phase, as well as decisions made throughout the residual hazards management period, are key to the long-term success of the prescribed system. In fact

  11. Casein kinase 2 down-regulation and activation by polybasic peptides are mediated by acidic residues in the 55-64 region of the beta-subunit. A study with calmodulin as phosphorylatable substrate

    DEFF Research Database (Denmark)

    Meggio, F; Boldyreff, B; Issinger, O G

    1994-01-01

    are conversely ineffective. The latent "calmodulin kinase" activity of CK2 can also be specifically unmasked by a peptide (alpha[66-86]) reproducing a basic insert of the catalytic subunit. This effect is reversed by equimolar addition of a peptide (beta[55-71]) including the 55-64 acidic stretch of the beta......-subunit. Comparable polylysine stimulation was observed with the holoenzymes reconstituted with either beta wt or the beta mutants capable of assembling with the alpha-subunit, with the notable exception of those bearing Ala substitutions for acidic residues at positions 55, 57, and 59-61. These were nearly...... insensitive to 42 nM polylysine, which conversely promotes a more than 10-fold increase of calmodulin phosphorylation with wild-type beta.(ABSTRACT TRUNCATED AT 250 WORDS)...

  12. Larvicidal activity of ajowan ( Trachyspermum ammi ) and Peru balsam ( Myroxylon pereira ) oils and blends of their constituents against mosquito, Aedes aegypti , acute toxicity on water flea, Daphnia magna , and aqueous residue.

    Science.gov (United States)

    Seo, Seon-Mi; Park, Hye-Mi; Park, Il-Kwon

    2012-06-13

    This study evaluated the larvicidal activity of 20 plant essential oils and components from ajowan ( Trachyspermum ammi ) and Peru balsam ( Myroxylon pereira ) oils against the mosquito, Aedes aegypti . Of the 20 plant essential oils, ajowan and Peru balsam oils at 0.1 mg/mL exhibited 100 and 97.5% larval mortality, respectively. At this same concentration, the individual constituents, (+)-camphene, benzoic acid, thymol, carvacrol, benzyl benzonate, and benzyl trans-cinnamate, caused 100% mortality. The toxicity of blends of constituents identified in two active oils indicated that thymol and benzyl benzoate were major contributors to the larvicidal activity of the artificial blend. This study also tested the acute toxicity of these two active oils and their major constituents against the water flea, Daphnia magna . Peru balsam oil and benzyl trans-cinnamate were the most toxic to D. magna. Two days after the treatment, residues of ajowan and Peru balsalm oils in water were 36.2 and 85.1%, respectively. Less than 50% of benzyl trans-cinnamate and thymol were detected in the water at 2 days after treatment. The results show that the essential oils of ajowan and Peru balsam and some of their constituents have potential as botanical insecticides against Ae. aegypti mosquito larvae.

  13. Distinctive binding modes and inhibitory mechanisms of two peptidic inhibitors of urokinase-type plasminogen activator with isomeric P1 residues

    DEFF Research Database (Denmark)

    Jiang, Longguang; Zhao, Baoyu; Xu, Peng

    2015-01-01

    Abstract Two isomeric piperidine derivatives (meta and para isomers) were used as arginine mimics in the P1 position of a cyclic peptidic inhibitor (CPAYSRYLDC) of urokinase-type plasminogen activator. The two resulting cyclic peptides showed vastly different affinities (∼70 fold) to the target...

  14. Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382

    NARCIS (Netherlands)

    Yin, Huifang; Pijning, Tjaard; Meng, Xiangfeng; Dijkhuizen, Lubbert; van Leeuwen, Sander Sebastiaan

    2017-01-01

    The β-galactosidase enzyme from Bacillus circulans ATCC 31382 BgaD is widely used in the food industry to produce prebiotic galactooligosaccharides (GOS). Recently, the crystal structure of a C-terminally truncated version of the enzyme (BgaD-D) has been elucidated. The roles of active site amino

  15. Multiple tyrosine residues in the intracellular domain of the common ß subunit of the interleukin 5 receptor are involved in activation of STAT5

    NARCIS (Netherlands)

    Dijk, Thamar B. van; Caldenhoven, Eric; Raaijmakers, J.A.M.; Lammers, J.W.J.; Koenderman, L.; Groot, Rolf P. de

    1997-01-01

    Abstract In contrast to the general model of cytokine-induced JAK/STAT signaling, tyrosine phosphorylation of the IL-5R ß chain seems to be dispensable for STAT activation in cells overexpressing exogenous STAT proteins. In this study we expressed IL-5 receptor mutants in 293 cells and studied

  16. Identification of tyrosine residues in the intracellular domain of the growth hormone receptor required for transcriptional signaling and Stat5 activation

    DEFF Research Database (Denmark)

    Hansen, L. H.; Wang, X.; Kopchick, J J

    1996-01-01

    The binding of growth hormone (GH) to its receptor results in its dimerization followed by activation of Jak2 kinase and tyrosine phosphorylation of the GH receptor itself, as well as Jak2 and the transcription factors Stat1, -3, and -5. In order to study the role of GH receptor tyrosine phosphor...

  17. Mass spectrometry and site-directed mutagenesis identify several autophosphorylated residues required for the activity of PrkC, a Ser/Thr kinase from Bacillus subtilis

    DEFF Research Database (Denmark)

    Madec, Edwige; Stensballe, Allan; Kjellström, Sven

    2003-01-01

    the kinase and a short juxtamembrane region. This fragment, which we designate PrkCc, undergoes autophosphorylation in E.coli. PrkCc is further autophosphorylated in vitro, apparently through a trans-kinase, intermolecular reaction. PrkC also displays kinase activity with myelin basic protein. Using high...

  18. Comparative Study of Microbial Activity and Chemical Properties of Soil by Implementing Anti-erosion Measure Vertical Mulching with Organic Residues

    Directory of Open Access Journals (Sweden)

    Gergana Slavova Kuncheva

    2015-01-01

    Full Text Available Water soil erosion is a phenomenon in which soil particles are separated transported and translocated by the action of rain water. Removal of topsoil by water flow leads to a decrease of humus in the soil, deterioration of soil structure, compaction, and reduction of microbial activity.Developed and tested have been number of methods and technologies for soil protection from the effects of water erosion. Such technology is vertical mulching, and straw or compost applied as mulching material.This work is a study of the changes that occur in some soil chemical properties and soil microbiological activity, as a result in the implementation of anti-erosion measure vertical mulching with different mulching materials for growing corn and wheat grain on carbonate chernozem, on sloping agricultural lands.

  19. A novel familial mutation in the PCSK1 gene that alters the oxyanion hole residue of proprotein convertase 1/3 and impairs its enzymatic activity.

    Directory of Open Access Journals (Sweden)

    Michael Wilschanski

    Full Text Available Four siblings presented with congenital diarrhea and various endocrinopathies. Exome sequencing and homozygosity mapping identified five regions, comprising 337 protein-coding genes that were shared by three affected siblings. Exome sequencing identified a novel homozygous N309K mutation in the proprotein convertase subtilisin/kexin type 1 (PCSK1 gene, encoding the neuroendocrine convertase 1 precursor (PC1/3 which was recently reported as a cause of Congenital Diarrhea Disorder (CDD. The PCSK1 mutation affected the oxyanion hole transition state-stabilizing amino acid within the active site, which is critical for appropriate proprotein maturation and enzyme activity. Unexpectedly, the N309K mutant protein exhibited normal, though slowed, prodomain removal and was secreted from both HEK293 and Neuro2A cells. However, the secreted enzyme showed no catalytic activity, and was not processed into the 66 kDa form. We conclude that the N309K enzyme is able to cleave its own propeptide but is catalytically inert against in trans substrates, and that this variant accounts for the enteric and systemic endocrinopathies seen in this large consanguineous kindred.

  20. Agrochemicals and Residues Newsletter. No. 6

    International Nuclear Information System (INIS)

    1991-09-01

    This newsletter provides very brief summaries of the current coordinated research programs and of the completed coordinated research program on the study of the biological activity and bioavailability of bound pesticide residues using nuclear techniques (a more complete report will be published in the IAEA Panel Proceedings series). Research in progress at the Agrochemicals Unit of the IAEA Laboratory in Seibersdorf is also described

  1. QA/QC in pesticide residue analysis

    International Nuclear Information System (INIS)

    Ambrus, A.

    2002-01-01

    This paper outlines problems related to pesticide residue analysis in a regulatory laboratory that are related to: availability of reference materials, as over 1000 pesticide active ingredients are currently in use and over 400 crops represent a large part of a healthy diet; analysis time; availability of samples in sufficient numbers; uncertainties of the procedures

  2. Quadratic residues and non-residues selected topics

    CERN Document Server

    Wright, Steve

    2016-01-01

    This book offers an account of the classical theory of quadratic residues and non-residues with the goal of using that theory as a lens through which to view the development of some of the fundamental methods employed in modern elementary, algebraic, and analytic number theory. The first three chapters present some basic facts and the history of quadratic residues and non-residues and discuss various proofs of the Law of Quadratic Reciprosity in depth, with an emphasis on the six proofs that Gauss published. The remaining seven chapters explore some interesting applications of the Law of Quadratic Reciprocity, prove some results concerning the distribution and arithmetic structure of quadratic residues and non-residues, provide a detailed proof of Dirichlet’s Class-Number Formula, and discuss the question of whether quadratic residues are randomly distributed. The text is a valuable resource for graduate and advanced undergraduate students as well as for mathematicians interested in number theory.

  3. Residual viraemia in HIV-1-infected patients with plasma viral load activation markers

    DEFF Research Database (Denmark)

    Ostrowski, S R; Katzenstein, T L; Pedersen, B K

    2008-01-01

    -count, CD4+HLA-DR+, CD8+HLA-DR+CD38+, CD4+CD45RA-CD45RO+, CD8+CD45RA-CD45RO+, CD4+CD45RA+CD62L+, CD8+CD45RA+CD62L+ T cells, IgG or IgM. In conclusion, RV was associated with increased blood levels of soluble immune activation markers in HAART-treated HIV-1-infected patients. The finding that RV...

  4. Acylated mono-, bis- and tris- Cinchona-Based Amines Containing Ferrocene or Organic Residues: Synthesis, Structure and in Vitro Antitumor Activity on Selected Human Cancer Cell Lines

    OpenAIRE

    Károlyi, Benedek Imre; Bősze, Szilvia; Orbán, Erika; Sohár, Pál; Drahos, László; Gál, Emese; Csámpai, Antal

    2012-01-01

    A series of novel functionalized mono-, bis- and tris-(S)-{[(2S,4R,8R)-8-ethyl-quinuclidin-2-yl](6-methoxyquinolin-4-yl)}methanamines including ferrocene-containing derivatives was obtained by the reaction of the precursor amine with a variety of acylation agents. Their in vitro antitumor activity was investigated against human leukemia (HL-60), human neuroblastoma (SH-SY5Y), human hepatoma (HepG2) and human breast cancer (MCF-7) cells by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoli...

  5. Redox Properties of Human Medium-Chain Acyl-CoA Dehydrogenase, Modulation by Charged Active-Site Amino Acid Residues

    OpenAIRE

    Mancini-Samuelson, Gina J.; Kieweg, Volker; Sabaj, Kim Marie; Ghisla, Sandro; Stankovich, Marian T.

    1998-01-01

    The modulation of the electron-transfer properties of human medium-chain acyl-CoA dehydrogenase (hwtMCADH) has been studied using wild-type and site-directed mutants by determining their midpoint potentials at various pH values and estimating the involved pKs. The mutants used were E376D, in which the negative charge is retained; E376Q, in which one negative charge (pKa ≈ 6.0) is removed from the active center; E99G, in which a different negative charge (pKa ≈ 7.3) also is affected; and E376H...

  6. Identification and characterization of hydrophobic gate residues in TRP channels.

    Science.gov (United States)

    Zheng, Wang; Hu, Ruikun; Cai, Ruiqi; Hofmann, Laura; Hu, Qiaolin; Fatehi, Mohammad; Long, Wentong; Kong, Tim; Tang, Jingfeng; Light, Peter; Flockerzi, Veit; Cao, Ying; Chen, Xing-Zhen

    2018-02-01

    Transient receptor potential (TRP) channels, subdivided into 6 subfamilies in mammals, have essential roles in sensory physiology. They respond to remarkably diverse stimuli, comprising thermal, chemical, and mechanical modalities, through opening or closing of channel gates. In this study, we systematically substituted the hydrophobic residues within the distal fragment of pore-lining helix S6 with hydrophilic residues and, based on Xenopus oocyte and mammalian cell electrophysiology and a hydrophobic gate theory, identified hydrophobic gates in TRPV6/V5/V4/C4/M8. We found that channel activity drastically increased when TRPV6 Ala616 or Met617 or TRPV5 Ala576 or Met577 , but not any of their adjacent residues, was substituted with hydrophilic residues. Channel activity strongly correlated with the hydrophilicity of the residues at those sites, suggesting that consecutive hydrophobic residues TRPV6 Ala616-Met617 and TRPV5 Ala576-Met577 form a double-residue gate in each channel. By the same strategy, we identified a hydrophobic single-residue gate in TRPV4 Iso715 , TRPC4 Iso617 , and TRPM8 Val976 . In support of the hydrophobic gate theory, hydrophilic substitution at the gate site, which removes the hydrophobic gate seal, substantially increased the activity of TRP channels in low-activity states but had little effect on the function of activated channels. The double-residue gate channels were more sensitive to small changes in the gate's hydrophobicity or size than single-residue gate channels. The unconventional double-reside gating mechanism in TRP channels may have been evolved to respond especially to physiologic stimuli that trigger relatively small gate conformational changes.-Zheng, W., Hu, R., Cai, R., Hofmann, L., Hu, Q., Fatehi, M., Long, W., Kong, T., Tang, J., Light, P., Flockerzi, V., Cao, Y., Chen, X.-Z. Identification and characterization of hydrophobic gate residues in TRP channels.

  7. Comparative studies on adsorptive removal of heavy metal ions by biosorbent, bio-char and activated carbon obtained from low cost agro-residue.

    Science.gov (United States)

    Kırbıyık, Çisem; Pütün, Ayşe Eren; Pütün, Ersan

    2016-01-01

    In this study, Fe(III) and Cr(III) metal ion adsorption processes were carried out with three adsorbents in batch experiments and their adsorption performance was compared. These adsorbents were sesame stalk without pretreatment, bio-char derived from thermal decomposition of biomass, and activated carbon which was obtained from chemical activation of biomass. Scanning electron microscopy and Fourier transform-infrared techniques were used for characterization of adsorbents. The optimum conditions for the adsorption process were obtained by observing the influences of solution pH, adsorbent dosage, initial solution concentration, contact time and temperature. The optimum adsorption efficiencies were determined at pH 2.8 and pH 4.0 for Fe(III) and Cr(III) metal ion solutions, respectively. The experimental data were modelled by different isotherm models and the equilibriums were well described by the Langmuir adsorption isotherm model. The pseudo-first-order, pseudo-second-order kinetic, intra-particle diffusion and Elovich models were applied to analyze the kinetic data and to evaluate rate constants. The pseudo-second-order kinetic model gave a better fit than the others. The thermodynamic parameters, such as Gibbs free energy change ΔG°, standard enthalpy change ΔH° and standard entropy change ΔS° were evaluated. The thermodynamic study showed the adsorption was a spontaneous endothermic process.

  8. Acylated mono-, bis- and tris- cinchona-based amines containing ferrocene or organic residues: synthesis, structure and in vitro antitumor activity on selected human cancer cell lines.

    Science.gov (United States)

    Károlyi, Benedek Imre; Bösze, Szilvia; Orbán, Erika; Sohár, Pál; Drahos, László; Gál, Emese; Csámpai, Antal

    2012-02-24

    A series of novel functionalized mono-, bis- and tris-(S)-{[(2S,4R,8R)-8-ethyl-quinuclidin-2-yl](6-methoxyquinolin-4-yl)}methanamines including ferrocene-containing derivatives was obtained by the reaction of the precursor amine with a variety of acylation agents. Their in vitro antitumor activity was investigated against human leukemia (HL-60), human neuroblastoma (SH-SY5Y), human hepatoma (HepG2) and human breast cancer (MCF-7) cells by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-assay and the 50% inhibitory concentration (IC(50)) values were determined. Our data indicate that the precursor amine has no antitumor activity in vitro, but the bis-methanamines with ureido-, thioureido and amide-type linkers display attractive in vitro cytotoxicity and cytostatic effects on HL-60, HepG2, MCF-7 and SH-SY5Y cells. Besides 1H- and 13C-NMR methods the structures of the new model compounds were also studied by DFT calculations.

  9. Acylated mono-, bis- and tris- Cinchona-Based Amines Containing Ferrocene or Organic Residues: Synthesis, Structure and in Vitro Antitumor Activity on Selected Human Cancer Cell Lines

    Directory of Open Access Journals (Sweden)

    Emese Gál

    2012-02-01

    Full Text Available A series of novel functionalized mono-, bis- and tris-(S-{[(2S,4R,8R-8-ethyl-quinuclidin-2-yl](6-methoxyquinolin-4-yl}methanamines including ferrocene-containing derivatives was obtained by the reaction of the precursor amine with a variety of acylation agents. Their in vitro antitumor activity was investigated against human leukemia (HL-60, human neuroblastoma (SH-SY5Y, human hepatoma (HepG2 and human breast cancer (MCF-7 cells by the 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT-assay and the 50% inhibitory concentration (IC50 values were determined. Our data indicate that the precursor amine has no antitumor activity in vitro, but the bis-methanamines with ureido-, thioureido and amide-type linkers display attractive in vitro cytotoxicity and cytostatic effects on HL-60, HepG2, MCF-7 and SH-SY5Y cells. Besides 1H- and 13C-NMR methods the structures of the new model compounds were also studied by DFT calculations.

  10. Redox properties of human medium-chain acyl-CoA dehydrogenase, modulation by charged active-site amino acid residues.

    Science.gov (United States)

    Mancini-Samuelson, G J; Kieweg, V; Sabaj, K M; Ghisla, S; Stankovich, M T

    1998-10-13

    The modulation of the electron-transfer properties of human medium-chain acyl-CoA dehydrogenase (hwtMCADH) has been studied using wild-type and site-directed mutants by determining their midpoint potentials at various pH values and estimating the involved pKs. The mutants used were E376D, in which the negative charge is retained; E376Q, in which one negative charge (pKa approximately 6. 0) is removed from the active center; E99G, in which a different negative charge (pKa approximately 7.3) also is affected; and E376H (pKa approximately 9.3) in which a positive charge is present. Em for hwtMCADH at pH 7.6 is -0.114 V. Results for the site-directed mutants indicate that loss of a negative charge in the active site causes a +0.033 V potential shift. This is consistent with the assumption that electrostatic interactions (as in the case of flavodoxins) and specific charges are important in the modulation of the electron-transfer properties of this class of dehydrogenases. Specifically, these charge interactions appear to correlate with the positive Em shift observed upon binding of substrate/product couple to MCADH [Lenn, N. D., Stankovich, M. T., and Liu, H. (1990) Biochemistry 29, 3709-3715], which coincides with a pK increase of Glu376-COOH from approximately 6 to 8-9 [Rudik, I., Ghisla, S., and Thorpe, C. (1998) Biochemistry 37, 8437-8445]. From the pH dependence of the midpoint potentials of hwtMCADH two mechanistically important ionizations are estimated. The pKa value of approximately 6.0 is assigned to the catalytic base, Glu376-COOH, in the oxidized enzyme based on comparison with the pH behavior of the E376H mutant, it thus coincides with the pK value recently estimated [Vock, P., Engst, S., Eder, M., and Ghisla, S. (1998) Biochemistry 37, 1848-1860]. The pKa of approximately 7.1 is assigned to Glu376-COOH in reduced hwtMCADH. Comparable values for these pKas for Glu376-COOH in pig kidney MCADH are pKox = 6.5 and pKred = 7.9. The Em measured for K304E-MCADH (a

  11. Bioenergy from sisal residues

    Energy Technology Data Exchange (ETDEWEB)

    Jungersen, G. [Dansk Teknologisk Inst. (Denmark); Kivaisi, A.; Rubindamayugi, M. [Univ. of Dar es Salaam (Tanzania, United Republic of)

    1998-05-01

    The main objectives of this report are: To analyse the bioenergy potential of the Tanzanian agro-industries, with special emphasis on the Sisal industry, the largest producer of agro-industrial residues in Tanzania; and to upgrade the human capacity and research potential of the Applied Microbiology Unit at the University of Dar es Salaam, in order to ensure a scientific and technological support for future operation and implementation of biogas facilities and anaerobic water treatment systems. The experimental work on sisal residues contains the following issues: Optimal reactor set-up and performance; Pre-treatment methods for treatment of fibre fraction in order to increase the methane yield; Evaluation of the requirement for nutrient addition; Evaluation of the potential for bioethanol production from sisal bulbs. The processing of sisal leaves into dry fibres (decortication) has traditionally been done by the wet processing method, which consumes considerable quantities of water and produces large quantities of waste water. The Tanzania Sisal Authority (TSA) is now developing a dry decortication method, which consumes less water and produces a waste product with 12-15% TS, which is feasible for treatment in CSTR systems (Continously Stirred Tank Reactors). (EG)

  12. Regulatory framework for NORM residues in Belgium

    Energy Technology Data Exchange (ETDEWEB)

    Pepin, S.; Dehandschutter, B.; Poffijn, A.; Sonck, M. [Federal Agency for Nuclear Control (FANC), Rue Ravenstein 36, 1000 Brussels (Belgium)

    2013-07-01

    The Belgian radiation protection authority (Federal Agency for Nuclear Control - FANC) has published in March 2013 a decree regulating the acceptance of NORM residues by nonradioactive waste treatment facilities. This regulation is based on the concept of 'work activities involving natural radiation sources' in the sense of article 40 of the 96/29/EURATOM directive. The disposal or processing facilities which accept NORM residues with an activity concentration above a generic exemption level will be considered as 'work activities' and submitted to declaration according to the Belgian radiation protection regulations. On basis of this declaration, specific acceptance criteria for the different types of processing/ disposal of the residues (disposal on landfill, recycling into building materials, etc.) are imposed. FANC has drafted guidelines for these acceptance criteria. A methodological guide for the operators of the concerned facilities was also published. Moreover, sites where significant quantities of NORM residues are or have been disposed, are subjected to an environmental monitoring in the framework of the national program of radiological surveillance of FANC. FANC also introduced in its regulations the concept of anthropogenic radon-prone areas: e.g. former phosphogypsum stacks have been defined as anthropogenic radon-prone areas, which allows some form of regulatory control of these sites. (authors)

  13. Comparison of residual stress in martensitic alloys by nondestructive techniques

    International Nuclear Information System (INIS)

    Roy, A.K.; Bandyopadhyay, S.; Suresh, S.B.; Wells, D.

    2006-01-01

    Three martensitic materials, namely Alloys EP-823, HT-9 and 422 were subjected to tensile loading at ambient temperature. The cylindrical specimens tested at different levels of tensile loading were analyzed for characterization of residual stress resulting from plastic deformation corresponding to the applied loads between the yield strength and the ultimate tensile strength. The extent of residual stress developed at these applied stresses was analyzed by nondestructive positron annihilation spectroscopy, activation, and neutron diffraction techniques. The results indicate that the residual stresses characterized by all three techniques exhibited a consistent pattern showing a gradual enhancement in residual stress with increasing applied load

  14. Residual Wage Differences by Gender: Bounding the Estimates.

    Science.gov (United States)

    Sakellariou, Chris N.; Patrinos, Harry A.

    1996-01-01

    Uses data from the 1986 Canadian labor market activity survey file to derive estimates of residual gender wage gap differences. Investigates these estimates' dependence on experimental design and on assumptions about discrimination-free wage structures. Residual differences persist, even after restricting the sample to a group of highly motivated,…

  15. The role of substrate binding pocket residues phenylalanine 176 and phenylalanine 196 on Pseudomonas sp. OX1 toluene o-xylene monooxygenase activity and regiospecificity.

    Science.gov (United States)

    Sönmez, Burcu; Yanık-Yıldırım, K Cansu; Wood, Thomas K; Vardar-Schara, Gönül

    2014-08-01

    Saturation mutagenesis was used to generate eleven substitutions of toluene-o-xylene monooxygenase (ToMO) at alpha subunit (TouA) positions F176 and F196 among which nine were novel: F176H, F176N, F176S, F176T, F196A, F196L, F196T, F196Y, F196H, F196I, and F196V. By testing the substrates phenol, toluene, and naphthalene, these positions were found to influence ToMO oxidation activity and regiospecificity. Specifically, TouA variant F176H was identified that had 4.7-, 4.3-, and 1.8-fold faster hydroxylation activity towards phenol, toluene, and naphthalene, respectively, compared to native ToMO. The F176H variant also produced the novel product hydroquinone (61%) from phenol, made twofold more 2-naphthol from naphthalene (34% vs. 16% by the wild-type ToMO), and had the regiospecificity of toluene changed from 51% to 73% p-cresol. The TouA F176N variant had the most para-hydroxylation capability, forming p-cresol (92%) from toluene and hydroquinone (82%) from phenol as the major product, whereas native ToMO formed 30% o-cresol, 19% m-cresol, and 51% of p-cresol from toluene and 100% catechol from phenol. For naphthalene oxidation, TouA variant F176S exhibited the largest shift in the product distribution by producing threefold more 2-naphthol. Among the other F196 variants, F196L produced catechol from phenol two times faster than the wild-type enzyme. The TouA F196I variant produced twofold less o-cresol and 19% more p-cresol from toluene, and the TouA F196A variant produced 62% more 2-naphthol from naphthalene compared to wild-type ToMO. Both of these positions have never been studied through the saturation mutagenesis and some of the best substitutions uncovered here have never been predicted and characterized for aromatics hydroxylation. © 2014 Wiley Periodicals, Inc.

  16. Delineating residues for haemolytic activities of snake venom cardiotoxin 1 from Naja naja as probed by molecular dynamics simulations and in vitro validations.

    Science.gov (United States)

    Gorai, Biswajit; Sivaraman, Thirunavukkarasu

    2017-02-01

    Cardiotoxins (CTXs) are single polypeptide chain consisting of 59-62 amino acids with four disulfide bridges and globular proteins of simple β-sheet folds. The CTXs are one of principal toxic components causing haemolysis and damaging various cells and belong to three-finger toxin (TFT) superfamily of snake venoms. However, there is no natural or synthetic small molecular inhibitor to the protein toxins to date. In the present study, modes of interaction of cardiotoxin 1 (CTX1) from Indian cobra (Naja naja) with heterogeneous erythrocyte membrane (EM) model system have been extensively examined by using all-atom molecular dynamics (MD) simulations in near physiological conditions and comprehensive analyses of the MD data revealed two distinct principal regions ('head groove' and 'loop groove') of the protein toxin for establishing structural interactions with the EM system. Moreover, combined analyses of data from high-throughput virtual screening of NCI small molecular database, in vitro haemolytic assays for top-hits of the chemical compounds against crude venom of Naja naja and as well CTXs purified from the venom and pharmacokinetic examinations on the chemical compounds retarding haemolytic activities of CTXs suggested that Etidronic acid and Zoledronic acid are promising prototypic chemical inhibitors to CTXs of snake venoms. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Inexpensive, effective novel activated carbon fibers for sample cleanup: application to multipesticide residue analysis in food commodities using a QuEChERS method.

    Science.gov (United States)

    Singh, Shiv; Srivastava, Anshuman; Singh, Sheelendra Pratap

    2018-03-01

    Phenolic resin based activated carbon fibers (ACFs) were applied for the first time as a reversed-dispersive solid-phase extraction (r-DSPE) sorbent. A modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) method was applied to determine 26 pesticides (organophosphates, organochlorines, synthetic pyrethroids, and herbicides) in different complex matrices, including cauliflower, cucumber, banana, apple, wheat, and black gram. Different physicochemical characterization techniques were used to investigate the engineering and structural properties of the r-DSPE sorbent. All the chromatographic analyses were performed with a gas chromatograph equipped with an electron capture detector. The recoveries of all 26 pesticides were acceptable (70-120%), with relative standard deviations of less than 15%. The limit of detection and the limit of quantification were 1.13-5.48 ng/g and 3.42-16.60 ng/g, respectively. In the original QuEChERS method, primary secondary amine is extensively used as the r-DSPE sorbent in the cleanup process, but it is eightfold more expensive than the ACFs used in this study. Therefore, the modified QuEChERS method using ACFs during the cleanup process is more efficient, cheaper, and more robust to determine pesticides from different types of matrices, including vegetables, grains, and fruits, and ACFs could be used as a cost-effective alternative to primary secondary amine. Graphical Abstract Sample clean-up using PSA and ACF as r-DSPE sorbent in QuEChERS method.

  18. Naturally occuring radioactivity in residues of drinking water treatment

    International Nuclear Information System (INIS)

    Vornehm, C.; Mallick, R.

    2009-01-01

    In the course of a research project about 500 residues of drinking water treatment from approx. 400 water supply companies in Bavaria were investigated on naturally occurring radioactivity. For each residue the effective dose for workers was evaluated for each residue. The results show that increased activities, particularly of Radium-226, can be found in the material. The dose due to the exposure to the residues, which mostly result from the backwashing of filters, is below the reference value of 1 mSv/a, which can be used according to paragraph 97 of the German radiation protection standard. During the project the quantity of residues in Bavaria and the ways of their disposal were evaluated. In addition the relation between the amount of natural radioisotopes in the residues and the geological and hydrochemical conditions of the water catchment area was pointed out. (orig.)

  19. Production of Human Cu,Zn SOD with Higher Activity and Lower Toxicity in E. coli via Mutation of Free Cysteine Residues

    Directory of Open Access Journals (Sweden)

    Kun Zhang

    2017-01-01

    Full Text Available Although, as an antioxidant enzyme, human Cu,Zn superoxide dismutase 1 (hSOD1 can mitigate damage to cell components caused by free radicals generated by aerobic metabolism, large-scale manufacturing and clinical use of hSOD1 are still limited by the challenge of rapid and inexpensive production of high-quality eukaryotic hSOD1 in recombinant forms. We have demonstrated previously that it is a promising strategy to increase the expression levels of soluble hSOD1 so as to increase hSOD1 yields in E. coli. In this study, a wild-type hSOD1 (wtSOD1 and three mutant SOD1s (mhSOD1s, in which free cysteines were substituted with serine, were constructed and their expression in soluble form was measured. Results show that the substitution of Cys111 (mhSOD1/C111S increased the expression of soluble hSOD1 in E. coli whereas substitution of the internal Cys6 (mhSOD1/C6S decreased it. Besides, raised levels of soluble expression led to an increase in hSOD1 yields. In addition, mhSOD1/C111S expressed at a higher soluble level showed lower toxicity and stronger whitening and antiradiation activities than those of wtSOD1. Taken together, our data demonstrate that C111S mutation in hSOD1 is an effective strategy to develop new SOD1-associated reagents and that mhSOD1/C111S is a satisfactory candidate for large-scale production.

  20. Estudio de la actividad biologica de dos suelos de los tableros costeros del NE de Brasil enmendados con residuos agricolas: vinaza y torta de canã de azúcar Study of biological activity in northeast Brazil's coastal tableland soils treated with organic residue: stillage and sugar cane solid residue

    Directory of Open Access Journals (Sweden)

    Zelson Tenório

    2000-04-01

    control before and after liming. Greater biological activity was observed under 10 and 20 t ha-1 of sugar cane solid residue treatment. In both soils, 90 m³ ha-1 of stillage induced minimum biological activity, very close to control response. In no case was inhibition of biological activity due to treatment observed. After incubation, some soil parameters were characterized to associate sugar cane organic fertilizers with soil fertility.

  1. Role of the RNA polymerase α subunits in CII-dependent activation of the bacteriophage λ pE promoter: identification of important residues and positioning of the α C-terminal domains

    Science.gov (United States)

    Kedzierska, Barbara; Lee, David J.; Węgrzyn, Grzegorz; Busby, Stephen J. W.; Thomas, Mark S.

    2004-01-01

    The bacteriophage λ CII protein stimulates the activity of three phage promoters, pE, pI and paQ, upon binding to a site overlapping the –35 element at each promoter. Here we used preparations of RNA polymerase carrying a DNA cleavage reagent attached to specific residues in the C-terminal domain of the RNA polymerase α subunit (αCTD) to demonstrate that one αCTD binds near position –41 at pE, whilst the other αCTD binds further upstream. The αCTD bound near position –41 is oriented such that its 261 determinant is in close proximity to σ70. The location of αCTD in CII-dependent complexes at the pE promoter is very similar to that found at many activator-independent promoters, and represents an alternative configuration for αCTD at promoters where activators bind sites overlapping the –35 region. We also used an in vivo alanine scan analysis to show that the DNA-binding determinant of αCTD is involved in stimulation of the pE promoter by CII, and this was confirmed by in vitro transcription assays. We also show that whereas the K271E substitution in αCTD results in a drastic decrease in CII-dependent activation of pE, the pI and paQ promoters are less sensitive to this substitution, suggesting that the role of αCTD at the three lysogenic promoters may be different. PMID:14762211

  2. Reúso de águas residuárias da piscicultura e da suinocultura na irrigação da cultura da alface Reuse of wastewater from swine and fish activities in the lettuce culture

    Directory of Open Access Journals (Sweden)

    Dirceu Baumgartner

    2007-04-01

    Full Text Available Este trabalho teve o objetivo de avaliar o desenvolvimento, a produção e a qualidade sanitária da cultura da alface irrigada com águas residuárias originadas da suinocultura e da piscicultura. Os tratamentos avaliados foram: T1 - alface irrigada com água de origem subterrânea e adubação suplementar; T2 - alface irrigada com água residuária de viveiro de peixes alimentados com ração; T3 - alface irrigada com água residuária originária de lagoa de estabilização de dejetos de suínos, e T4 - água de lagoa de cultivo de algas, alimentada com resíduo de biodigestor de dejeto de suíno. Os tratamentos não apresentaram diferenças significativas para altura da alface, diâmetro da cabeça, comprimento da raiz, massa da raiz, massa total da planta, massa fresca e massa seca; os tratamentos T4 e T2 apresentaram os maiores valores para comprimento da maior folha e número de folhas, respectivamente; em geral, as análises foliares e a extração de macro e micronutrientes pela cultura não apresentaram diferenças significativas; ocorreu contaminação de coliformes fecais e totais em todos os tratamentos; não ocorreu contaminação de Escherichia coli em todos os tratamentos; todos os Coeficientes de Uniformidade de Christiansen (CUC encontrados foram acima de 85%, exceto para o tratamento T3 que foi de 74,05%; ocorreram alterações químicas no solo proporcionais às características das águas utilizadas nos respectivos tratamentos.The objective of this work was to evaluate development, production and sanitary quality of the irrigated lettuce with wastewater from fish and swine activities. The evaluated treatments was: T1 - lettuce irrigated with water from underground origin and supplemental fertilization; T2 - lettuce irrigated with wastewater from fish activities and fish fed with ration; T3 - lettuce irrigated with wastewater from swine treated in stabilization lagoon; and T4 - lettuce irrigated with effluent from seaweed

  3. Functional significance of the conserved residues for the 23-residue module among MTH1 and MutT family proteins.

    Science.gov (United States)

    Fujii, Y; Shimokawa, H; Sekiguchi, M; Nakabeppu, Y

    1999-12-31

    Human MTH1 and Escherichia coli MutT proteins hydrolyze 7, 8-dihydro-8-oxo-dGTP (8-oxo-dGTP) to monophosphate, thus avoiding the incorporation of 8-oxo-7,8-dihydroguanine into nascent DNA. Although only 30 amino acid residues (23%) are identical between MTH1 and MutT, there is a highly conserved region consisting of 23 residues (MTH1, Gly(36)-Gly(58)) with 14 identical residues. A chimeric protein MTH1-Ec, in which the 23-residue sequence of MTH1 was replaced with that of MutT, retains its capability to hydrolyze 8-oxo-dGTP, thereby indicating that the 23-residue sequences of MTH1 and MutT are functionally and structurally equivalent and constitute functional modules. By saturation mutagenesis of the module in MTH1, 14 of the 23 residues proved to be essential to exert 8-oxo-dGTPase activity. For the other 9 residues (40, 42, 44, 46, 47, 49, 50, 54, and 58), positive mutants were obtained, and Arg(50) can be replaced with hydrophobic residues (Val, Leu, or Ile), with a greater stability and higher specific activity of the enzyme. Indispensabilities of Val(39), Ile(45), and Leu(53) indicate that an amphipathic property of alpha-helix I consisting of 14 residues of the module (Thr(44)-Gly(58)) is essential to maintain the stable catalytic surface for 8-oxo-dGTPase.

  4. Forest residues in cattle feed

    Directory of Open Access Journals (Sweden)

    João Elzeário Castelo Branco Iapichini

    2012-12-01

    Full Text Available The ruminants are capable of converting low-quality food, when they are complementes with high-energy source. Through the use of regional agricultural residues is possible to conduct more economical production systems, since energetic foods have high cost in animal production. There is very abundant availability of residues in agroforestry activities worldwide, so that if a small fraction of them were used with appropriate technical criteria they could largely meet the needs of existing herds in the world and thus meet the demands of consumption of protein of animal origin. The Southwest Region of São Paulo State has large area occupied by reforestation and wide availability of non-timber forest residues, which may represent more concentrated energetic food for ruminant production. This experiment aimed to evaluate the acceptability of ground pine (20, 30 and 40%, replacing part of the energetic food (corn, present in the composition of the concentrate and was performed at the Experimental Station of Itapetininga - Forest Institute / SMA, in the dry season of 2011. It were used four crossbred steers, mean 18 months old, average body weight of 250 kg, housed in a paddock provided with water ad libitum and covered troughs for supplementation with the experimental diet. The adjustment period of the animals was of 07 days and the measurement of the levels of consumption, physiological changes, acceptability and physiological parameters were observed during the following 25 days. The concentrate supplement was formulated based on corn (76.2%, Soybean Meal (20%, urea (2%, Ammonium sulfate (0.4%, calcite (1.4%, Mineral Core (1% and finely ground Pine Cone, replacing corn. In preparing food, the formulas were prepared to make them isoproteic/energetic, containing the following nutrient levels: 22% Crude Protein (CP and 79% of Total Nutrients (TDN. The animals received the supplement in three steps for each level of cone replaced, being offered in the

  5. Discovery of [ cis-3-({(5 R)-5-[(7-Fluoro-1,1-dimethyl-2,3-dihydro-1 H-inden-5-yl)carbamoyl]-2-methoxy-7,8-dihydro-1,6-naphthyridin-6(5 H)-yl}carbonyl)cyclobutyl]acetic Acid (TAK-828F) as a Potent, Selective, and Orally Available Novel Retinoic Acid Receptor-Related Orphan Receptor γt Inverse Agonist.

    Science.gov (United States)

    Kono, Mitsunori; Ochida, Atsuko; Oda, Tsuneo; Imada, Takashi; Banno, Yoshihiro; Taya, Naohiro; Masada, Shinichi; Kawamoto, Tetsuji; Yonemori, Kazuko; Nara, Yoshi; Fukase, Yoshiyuki; Yukawa, Tomoya; Tokuhara, Hidekazu; Skene, Robert; Sang, Bi-Ching; Hoffman, Isaac D; Snell, Gyorgy P; Uga, Keiko; Shibata, Akira; Igaki, Keiko; Nakamura, Yoshiki; Nakagawa, Hideyuki; Tsuchimori, Noboru; Yamasaki, Masashi; Shirai, Junya; Yamamoto, Satoshi

    2018-03-14

    A series of tetrahydronaphthyridine derivatives as novel RORγt inverse agonists were designed and synthesized. We reduced the lipophilicity of tetrahydroisoquinoline compound 1 by replacement of the trimethylsilyl group and SBDD-guided scaffold exchange, which successfully afforded compound 7 with a lower log  D value and tolerable in vitro activity. Consideration of LLE values in the subsequent optimization of the carboxylate tether led to the discovery of [ cis-3-({(5 R)-5-[(7-fluoro-1,1-dimethyl-2,3-dihydro-1 H-inden-5-yl)carbamoyl]-2-methoxy-7,8-dihydro-1,6-naphthyridin-6(5 H)-yl}carbonyl)cyclobutyl]acetic acid, TAK-828F (10), which showed potent RORγt inverse agonistic activity, excellent selectivity against other ROR isoforms and nuclear receptors, and a good pharmacokinetic profile. In animal studies, oral administration of compound 10 exhibited robust and dose-dependent inhibition of IL-17A cytokine expression in a mouse IL23-induced gene expression assay. Furthermore, development of clinical symptoms in a mouse experimental autoimmune encephalomyelitis model was significantly reduced. Compound 10 was selected as a clinical compound for the treatment of Th17-driven autoimmune diseases.

  6. Identification of Key Residues for Enzymatic Carboxylate Reduction

    Directory of Open Access Journals (Sweden)

    Holly Stolterfoht

    2018-02-01

    Full Text Available Carboxylate reductases (CARs, E.C. 1.2.1.30 generate aldehydes from their corresponding carboxylic acid with high selectivity. Little is known about the structure of CARs and their catalytically important amino acid residues. The identification of key residues for carboxylate reduction provides a starting point to gain deeper understanding of enzymatic carboxylate reduction. A multiple sequence alignment of CARs with confirmed activity recently identified in our lab and from the literature revealed a fingerprint of conserved amino acids. We studied the function of conserved residues by multiple sequence alignments and mutational replacements of these residues. In this study, single-site alanine variants of Neurospora crassa CAR were investigated to determine the contribution of conserved residues to the function, expressability or stability of the enzyme. The effect of amino acid replacements was investigated by analyzing enzymatic activity of the variants in vivo and in vitro. Supported by molecular modeling, we interpreted that five of these residues are essential for catalytic activity, or substrate and co-substrate binding. We identified amino acid residues having significant impact on CAR activity. Replacement of His 237, Glu 433, Ser 595, Tyr 844, and Lys 848 by Ala abolish CAR activity, indicating their key role in acid reduction. These results may assist in the functional annotation of CAR coding genes in genomic databases. While some other conserved residues decreased activity or had no significant impact, four residues increased the specific activity of NcCAR variants when replaced by alanine. Finally, we showed that NcCAR wild-type and mutants efficiently reduce aliphatic acids.

  7. Identification of Key Residues for Enzymatic Carboxylate Reduction.

    Science.gov (United States)

    Stolterfoht, Holly; Steinkellner, Georg; Schwendenwein, Daniel; Pavkov-Keller, Tea; Gruber, Karl; Winkler, Margit

    2018-01-01

    Carboxylate reductases (CARs, E.C. 1.2.1.30) generate aldehydes from their corresponding carboxylic acid with high selectivity. Little is known about the structure of CARs and their catalytically important amino acid residues. The identification of key residues for carboxylate reduction provides a starting point to gain deeper understanding of enzymatic carboxylate reduction. A multiple sequence alignment of CARs with confirmed activity recently identified in our lab and from the literature revealed a fingerprint of conserved amino acids. We studied the function of conserved residues by multiple sequence alignments and mutational replacements of these residues. In this study, single-site alanine variants of Neurospora crassa CAR were investigated to determine the contribution of conserved residues to the function, expressability or stability of the enzyme. The effect of amino acid replacements was investigated by analyzing enzymatic activity of the variants in vivo and in vitro . Supported by molecular modeling, we interpreted that five of these residues are essential for catalytic activity, or substrate and co-substrate binding. We identified amino acid residues having significant impact on CAR activity. Replacement of His 237, Glu 433, Ser 595, Tyr 844, and Lys 848 by Ala abolish CAR activity, indicating their key role in acid reduction. These results may assist in the functional annotation of CAR coding genes in genomic databases. While some other conserved residues decreased activity or had no significant impact, four residues increased the specific activity of Nc CAR variants when replaced by alanine. Finally, we showed that Nc CAR wild-type and mutants efficiently reduce aliphatic acids.

  8. Marine Tar Residues: a Review

    OpenAIRE

    Warnock, April M.; Hagen, Scott C.; Passeri, Davina L.

    2015-01-01

    Marine tar residues originate from natural and anthropogenic oil releases into the ocean environment and are formed after liquid petroleum is transformed by weathering, sedimentation, and other processes. Tar balls, tar mats, and tar patties are common examples of marine tar residues and can range in size from millimeters in diameter (tar balls) to several meters in length and width (tar mats). These residues can remain in the ocean environment indefinitely, decomposing or becoming buried in ...

  9. Atividade residual in vitro do pelo de cobertura de cães tratados com dinotefuran sobre larvas e adultos de Ctenocephalides felis felis (Bouché, 1835 (Siphonaptera: pulicidae Residual activity in vitro of treated dog's hair coat with dinotefuran on larvae and adults of Ctenocephalides felis felis (Bouché, 1835 (Siphonaptera: Pulicidae

    Directory of Open Access Journals (Sweden)

    Thaís R. Correia

    2008-12-01

    Full Text Available O objetivo deste trabalho foi avaliar a atividade in vitro do resíduo em pelo de cobertura de cães tratados com dinotefuran sobre larvas e adultos de Ctenocephalides felis felis. Foram utilizadas três cadelas da Raça Beagle: a nº 1 foi tratada com uma formulação spray de dinotefuran a 0,834%, a nº 2 com uma formulação "strip-on" de dinotefuran a 30% e a nº 3 foi mantida sem tratamento. Pequenas áreas foram tricotomizadas nos dias 2, 9, 16, 23, 30, 37 e 44 após o tratamento. Para avaliação adulticida e larvicida, foram utilizados pulgas de uma colônia mantida em laboratório. Seis repetições com cada uma contendo 10 exemplares de C. f. felis, acondicionados em tubos de ensaio, da etapa correspondente foram utilizados por dia de desafio. Em cada repetição foi adicionado 0,02 gramas de pelo de cada área tricotomizada e dieta na etapa de larvas. O material da etapa adulto foi avaliado num período de 24 horas, o da etapa larvas foi avaliado após 20 dias de cada desafio. O resíduo no pêlo de cães tratados com ambas as formulações de dinotefuran foi eficaz no controle de larvas por um período de 44 dias, e no controle de adultos as formulações spray e a "strip-on" foram eficazes por um período de 16 e 23 dias, respectivamente.The aim of this study was to evaluate the residual activity of treated dog's hair coat with dinotefuran on larvae and adults of Ctenocephalides felis felis. Three female Beagle dogs were used. One female dog was treated with 0.834% dinotefuran spray, the second was treated with 30% dinotefuran strip-on and the third was not treated. Some areas of dog's hair were clipped on days 2, 9, 16, 23, 30, 37 and 44 after treatment. For the evaluation of adulticidal and larvicidal activities, flea adults and larvae from the laboratory colony were used. Six repetitions were used with 10 samples of each flea stage per day, placed in assay tubes. In each repetition we added 0.02 g of treated or untreated dog

  10. Overexpression of constitutively active MAP3K7 in ameloblasts causes enamel defects of mouse teeth.

    Science.gov (United States)

    Jinping, Zhao; Qing, Chu; Wenying, Song; Chunyan, Yang; Lili, Xiang; Yao, Shi; Yumin, Wang; Zhenzhen, Xu; Li, Zhang; Yuguang, Gao

    2017-12-01

    Compelling evidence suggests that mitogen-activated protein kinases (Mapks) play an important role in amelogenesis. However, the role of transforming growth factor (TGF)-β-activating kinase 1 (Tak1, Map3k7), which is a known upstream kinase of Mapks, during amelogenesis remains to be determined. The aim of this study was to investigate the possible involvement of Map3k7 in amelogenesis. We generated transgenic mice that produced constitutively active human MAP3K7 (caMAP3K7) under the control of amelogenin (Amelx) gene promoter. Radiography and micro-computed tomography (μCT) analysis was used to detect the radio-opacity and density of the teeth. The enamel microstructure was observed with a scanning electron microscope. Histological analysis was used to observe the adhesion between ameloblasts and residual organic matrix of the enamel. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the expression of enamel matrix protein. The enamel of mandibular molars in caMAP3K7-overexpressing mice displayed pigmentation and a highly irregular structure compared with the wild type littermates. Teeth of transgenic animals underwent rapid attrition due to the brittleness of the enamel layer. The microstructure of enamel, normally a highly ordered arrangement of hydroxyapatite crystals, was completely disorganized. The gross histological appearances of ameloblasts and supporting cellular structures, as well as the expression of the enamel protein amelotin (Amtn) were altered by the overexpression of caMAP3K7. Our data demonstrated that protein expression, processing and secretion occurred abnormally in transgenic mice overexpressing caMAP3K7. The overexpression of caMAP3K7 had a profound effect on enamel structure by disrupting the orderly growth of enamel prisms. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Separation and preparation of 6-gingerol from molecular distillation residue of Yunnan ginger rhizomes by high-speed counter-current chromatography and the antioxidant activity of ginger oils in vitro.

    Science.gov (United States)

    Gan, Zhilin; Liang, Zheng; Chen, Xiaosong; Wen, Xin; Wang, Yuxiao; Li, Mo; Ni, Yuanying

    2016-02-01

    Molecular distillation residue (MD-R) from ginger had the most total phenol content of 247.6mg gallic acid equivalents per gram (GAE/g) among the ginger oils. High-speed counter-current chromatography (HSCCC) technique in semi-preparative scale was successfully performed in separation and purification of 6-gingerol from MD-R by using a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (10:2:5:7, v/v/v/v). The target compound was isolated, collected, purified by HSCCC in the head-tail mode, and then analyzed by HPLC. A total of 90.38±0.53mg 6-gingerol was obtained from 600mg MD-R, with purity of 99.6%. In addition, the structural identification of 6-gingerol was performed by EI/MS, (1)H NMR and (13)C NMR. Moreover, the orders of antioxidant activity were vitamin E (VE)>supercritical fluid extraction oleoresin (SFE-O)=MD-R=6-gingerol>molecular distillation essential oil (MD-EO) and butylated hydroxytoluene (BHT)=VE>6-gingerol>MD-R=SFE-O>MD-EO, respectively in 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) scavenging and β-Carotene bleaching. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Treatment of Explosives Residues From Range Activities

    Science.gov (United States)

    2010-01-01

    metal wire (steel wire heated till it was glowing orange in a bunsen burner ) were used as sources of ignition. These sources were meant to represent...laboratory conditions. Neither red-hot metal probes nor open flames resulted in ignition or smoldering.  Methods to efficiently mix and apply...soybean oil in a very dry and in a moderately wet state. Both an open flame (butane-type lighter used for lighting candles, barbecues, etc.) and a hot

  13. Atividade residual de herbicidas aplicados em pós-emergência na cultura da soja sobre o milheto cultivado em sucessão Effect of the residual activity of herbicides applied in post-emergence soybean on pearl millet grown in succession

    Directory of Open Access Journals (Sweden)

    H.A Dan

    2011-09-01

    Full Text Available O objetivo deste trabalho foi avaliar a atividade residual de herbicidas utilizados em pós-emergência da cultura da soja sobre o milheto cultivado em sucessão. O experimento foi realizado em Latossolo Vermelho distroférrico de textura argilosa em região de cerrado. Os herbicidas chlorimuron-ethyl (0,015 kg ha-1, imazethapyr (0,060 kg ha-1, imazethapyr (0,100 kg ha-1 e fomesafen (0,250 kg ha-1 foram utilizados em pós-emergência do cultivar de soja Msoy-6101. Utilizou-se o delineamento de blocos casualizados, com quatro repetições, em esquema de parcelas subdivididas (5 x 4. Nas subparcelas, realizou-se a semeadura do milheto (híbrido ADR-7010 em quatro períodos, correspondendo a 0, 40, 80 e 120 dias após a aplicação dos herbicidas (DAA. Durante a condução do ensaio, avaliou-se a intoxicação da cultura aos 7 e 28 dias após a emergência, o estande, a altura e a matéria seca da parte aérea das plantas de milheto. Ao final do ciclo da cultura, determinou-se o rendimento de grãos. O imazethapyr (0,060 kg ha-1 e chlorimuron-ethyl (0,015 kg ha-1 não alteraram significativamente o rendimento da cultura do milheto em semeaduras posteriores a 80 DAA. Para fomesafen, o intervalo mínimo de segurança entre a aplicação e a semeadura do milheto foi de 100 dias. Por outro lado, maior persistência foi observada para imazethapyr na dose 0,100 kg ha-1, chegando a 120 dias de bioatividade sobre o milheto, que teve seu rendimento de grãos alterando mesmo quando semeado durante esse período.The objective of this study was to evaluate the residual activity of herbicides used in post-emergence soybean on Pearl Millet grown in succession. The experiment was conducted in clay soil in cerrado region. The herbicides chlorimuron-ethyl (0.015 kg ha-1, imazethapyr (0.060 kg ha-1, imazethapyr (0.100 kg ha-1 and fomesafen (0.250 kg ha-1 were used in post-emergence soybean cultivar Msoy-6101, in plots of 80 m² that were subdivided into sub-plots of

  14. Determination of Pesticides Residues in Cucumbers Grown in Greenhouse and the Effect of Some Procedures on Their Residues.

    Science.gov (United States)

    Leili, Mostafa; Pirmoghani, Amin; Samadi, Mohammad Taghi; Shokoohi, Reza; Roshanaei, Ghodratollah; Poormohammadi, Ali

    2016-11-01

    The objective of this study was to determine the residual concentrations of ethion and imidacloprid in cucumbers grown in greenhouse. The effect of some simple processing procedures on both ethion and imidacloprid residues were also studied. Ten active greenhouses that produce cucumber were randomly selected. Ethion and imidacloprid as the most widely used pesticides were measured in cucumber samples of studied greenhouses. Moreover, the effect of storing, washing, and peeling as simple processing procedures on both ethion and imidacloprid residues were investigated. One hour after pesticide application; the maximum residue levels (MRLs) of ethion and imidacloprid were higher than that of Codex standard level. One day after pesticide application, the levels of pesticides were decreased about 35 and 31% for ethion and imidacloprid, respectively, which still were higher than the MRL. Washing procedure led to about 51 and 42.5% loss in ethion and imidacloprid residues, respectively. Peeling procedure also led to highest loss of 93.4 and 63.7% in ethion and imidacloprid residues, respectively. The recovery for both target analytes was in the range between 88 and 102%. The residue values in collected samples one hour after pesticides application were higher than standard value. The storing, washing, and peeling procedures lead to the decrease of pesticide residues in greenhouse cucumbers. Among them, the peeling procedure has the greatest impact on residual reduction. Therefore, these procedures can be used as simple and effective processing techniques for reducing and removing pesticides from greenhouse products before their consumption.

  15. Landfilling of waste incineration residues

    DEFF Research Database (Denmark)

    Christensen, Thomas Højlund; Astrup, Thomas; Cai, Zuansi

    2002-01-01

    Residues from waste incineration are bottom ashes and air-pollution-control (APC) residues including fly ashes. The leaching of heavy metals and salts from the ashes is substantial and a wide spectrum of leaching tests and corresponding criteria have been introduced to regulate the landfilling...

  16. Atividade microbiana em solos, influenciada por resíduos de algodão e torta de mamona Microbial activity in soils influenced by residues of cotton and castor bean presscake

    Directory of Open Access Journals (Sweden)

    Silvia Capuani

    2012-12-01

    Full Text Available A atividade microbiana não se constitui apenas como bom indicador da qualidade do solo, mas é influenciada pela adição de carbono no sistema, o qual serve como substrato aos micro-organismos que aumentam sua atividade e a liberação de CO2, compreendendo a respiração edáfica do solo. Objetivou-se, neste trabalho, avaliar a atividade microbiana em diferentes tipos de solo com a adição de torta de mamona e resíduo têxtil de algodão. O trabalho foi conduzido em casa de vegetação na sede da Embrapa Algodão, em delineamento de blocos casualisados em esquema fatorial 4 x 3, com quatro repetições. A intervalos predeterminados de 4 dias os recipientes foram abertos e a solução de NaOH foi titulada com HCl 2 N na presença de indicador ácido/base fenolftaleína. Após a leitura a mesma quantidade de NaOH foi reposta e os recipientes novamente fechados. A diferença entre a quantidade de ácido necessária para neutralizar o hidróxido de sódio no recipiente testemunha e nos tratamentos equivale à quantidade de gás carbônico produzido pelos micro-organismos do solo. Constatou-se que os resíduos influenciaram significativamente a atividade microbiana nos diferentes tipos de solo, sobretudo nas primeiras determinações, apresentando-se como boas fontes para mineralização e fornecimento de nutrientes, tendo a torta de mamona proporcionado maior liberação acumulada de CO2 pelos micro-organismos.Microbial activity constitutes a good indicator of soil quality, and is influenced by the addition of carbon in the system serving as a substrate for microorganisms that increase their activity and release of CO2, comprising the edaphic respiration of the soil. The objective of this study was to evaluate the microbial activity in different soil types with the addition of cake press of castor bean and cotton textile residue. The study was conducted in a greenhouse at the headquarters of Embrapa Cotton in randomized block design in 4 x 3

  17. Statistical inference on residual life

    CERN Document Server

    Jeong, Jong-Hyeon

    2014-01-01

    This is a monograph on the concept of residual life, which is an alternative summary measure of time-to-event data, or survival data. The mean residual life has been used for many years under the name of life expectancy, so it is a natural concept for summarizing survival or reliability data. It is also more interpretable than the popular hazard function, especially for communications between patients and physicians regarding the efficacy of a new drug in the medical field. This book reviews existing statistical methods to infer the residual life distribution. The review and comparison includes existing inference methods for mean and median, or quantile, residual life analysis through medical data examples. The concept of the residual life is also extended to competing risks analysis. The targeted audience includes biostatisticians, graduate students, and PhD (bio)statisticians. Knowledge in survival analysis at an introductory graduate level is advisable prior to reading this book.

  18. Automatic prediction of catalytic residues by modeling residue structural neighborhood

    Directory of Open Access Journals (Sweden)

    Passerini Andrea

    2010-03-01

    Full Text Available Abstract Background Prediction of catalytic residues is a major step in characterizing the function of enzymes. In its simpler formulation, the problem can be cast into a binary classification task at the residue level, by predicting whether the residue is directly involved in the catalytic process. The task is quite hard also when structural information is available, due to the rather wide range of roles a functional residue can play and to the large imbalance between the number of catalytic and non-catalytic residues. Results We developed an effective representation of structural information by modeling spherical regions around candidate residues, and extracting statistics on the properties of their content such as physico-chemical properties, atomic density, flexibility, presence of water molecules. We trained an SVM classifier combining our features with sequence-based information and previously developed 3D features, and compared its performance with the most recent state-of-the-art approaches on different benchmark datasets. We further analyzed the discriminant power of the information provided by the presence of heterogens in the residue neighborhood. Conclusions Our structure-based method achieves consistent improvements on all tested datasets over both sequence-based and structure-based state-of-the-art approaches. Structural neighborhood information is shown to be responsible for such results, and predicting the presence of nearby heterogens seems to be a promising direction for further improvements.

  19. Residual magnetic field in rotary machines; Campo magnetico residual en maquinas rotatorias

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez V, Esteban A; Apanco R, Marcelino [Instituto de Investigaciones Electricas, Cuernavaca, Morelos (Mexico)

    2007-07-01

    The residual magnetism is a phenomenon in which the magnetic dipoles of a substance are oriented in a certain degree. On the other hand, when internal forces exist capable of aligning elementary magnetic dipoles of a material, a permanent magnet is obtained. Just as in a conductor or in a material, in the elements of a rotary electrical machine magnetic fields can be induced that produce a residual magnetism or magnetization. In the rotary electrical machines, the magnetization phenomenon causes serious problems, such as the generation of induced currents that propitiate the mechanical wear in bearings, collars, trunnions and inclusive in the shaft, by effects known as pitting, frosting and spark tracks, as well as erroneous readings in vibration and temperature sensors, that in some cases can cause the shut down of the machine. In this article are presented the general concepts on the residual magnetism in rotary electrical machines, the causes that originate it and the problems that arises, as well as the demagnetization of the components that have residual magnetic field. The results obtained by the area of Electrical Equipment of the Instituto de Investigaciones Electricas are revised, during the execution of activities related to the measurement and elimination of the residual magnetic field in rotary electrical machines. [Spanish] El magnetismo residual es un fenomeno en el que los dipolos magneticos de una sustancia se encuentran orientados en un grado determinado. Por otro lado, cuando existen fuerzas internas capaces de alinear los dipolos magneticos elementales de un material, se tiene un iman permanente. Al igual que en un conductor o un material, en los elementos de una maquina electrica rotatoria se pueden inducir campos magneticos que producen un magnetismo residual o magnetizacion. En las maquinas electricas rotatorias, el fenomeno de magnetizacion causa graves problemas, como la generacion de corrientes inducidas que propician el desgaste mecanico

  20. Economic Modeling of Residual Generation for the Lingayen Gulf Watershed

    Directory of Open Access Journals (Sweden)

    Douglas McGlone

    2000-12-01

    Full Text Available The Philippines is one of four countries involved in the Southeast Asian core project of LOICZ (Land Ocean Interactions in the Coastal Zone which has among its general goals the determination of how changes in human activities affect the fluxes of materials between land, sea, and atmosphere through the coastal zone. The economic component of the Philippine project addresses the question: “How does a change in economic activity affect coastal waters?” Of particular concern is the introduction of anthropogenically-derived residuals (N, P, C, SS into coastal waters. A regional input-output (IO model for Region 1 of the Philippines has been developed to estimate how projected changes in economic activity may affect residual flows into Lingayen Gulf. A residual coefficient matrix, derived from information obtained with a rapid assessment model (RA of residual generation in the Lingayen Gulf watershed, has been incorporated into the IO model. Such a model allows for analysis of various economic scenarios for the region, with projections of residual generation as the output. The resulting changes in residual flows may then serve as inputs to biogeochemical models of Lingayen Gulf. From this process, the impact of various economic scenarios on the water quality of Lingayen Gulf may be ascertained.This paper discusses and compares the RA and IO models of residual generation for the Lingayen Gulf watershed and provides examples of the scenario analysis process.

  1. Residue management in the Bolivia-Brazil gas pipeline construction

    International Nuclear Information System (INIS)

    Freitas, Eduardo Lopes; Henrique, Paulo Roberto Pereira; Cantarino, Anderson Americo Alves

    2000-01-01

    The construction of the gas pipeline is a process sequential of assembly phases, where each one of those phases generates residues of the most varied types and amounts, being necessary the forecast of your generation in agreement with the activity that is being executed. During the accomplishment of the works they are generated a lot of times situations where are observed the inadequate disposition of the residues. Those practices, besides the environmental impact that they cause, it can cart in the future, the need of additional investments be proceeded in the recovery of the areas and removal of the residues. This work presents the Program of administration of Residues instituted during the construction of the pipeline Bolivia - Brazil, seeking, on a side to reduce to the minimum the generation of residues and of the other, moths handling guidelines and disposition, in way to minimize the environmental impacts caused by the same ones. (author)

  2. Residual stress by repair welds

    International Nuclear Information System (INIS)

    Mochizuki, Masahito; Toyoda, Masao

    2003-01-01

    Residual stress by repair welds is computed using the thermal elastic-plastic analysis with phase-t