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Sample records for pepper 26s proteasome

  1. Functional study of hot pepper 26S proteasome subunit RPN7 induced by Tobacco mosaic virus from nuclear proteome analysis

    International Nuclear Information System (INIS)

    Lee, Boo-Ja; Kwon, Sun Jae; Kim, Sung-Kyu; Kim, Ki-Jeong; Park, Chang-Jin; Kim, Young-Jin; Park, Ohkmae K.; Paek, Kyung-Hee

    2006-01-01

    Two-dimensional gel electrophoresis (2-DE) was applied for the screening of Tobacco mosaic virus (TMV)-induced hot pepper (Capsicum annuum cv. Bugang) nuclear proteins. From differentially expressed protein spots, we acquired the matched peptide mass fingerprint (PMF) data, analyzed by MALDI-TOF MS, from the non-redundant hot pepper EST protein FASTA database using the VEMS 2.0 software. Among six identified nuclear proteins, the hot pepper 26S proteasome subunit RPN7 (CaRPN7) was subjected to further study. The level of CaRPN7 mRNA was specifically increased during incompatible TMV-P 0 interaction, but not during compatible TMV-P 1.2 interaction. When CaRPN7::GFP fusion protein was targeted in onion cells, the nuclei had been broken into pieces. In the hot pepper leaves, cell death was exacerbated and genomic DNA laddering was induced by Agrobacterium-mediated transient overexpression of CaPRN7. Thus, this report presents that the TMV-induced CaRPN7 may be involved in programmed cell death (PCD) in the hot pepper plant

  2. 26 S proteasomes function as stable entities

    DEFF Research Database (Denmark)

    Hendil, Klavs B; Hartmann-Petersen, Rasmus; Tanaka, Keiji

    2002-01-01

    , shuttles between a free state and the 26-S proteasome, bringing substrate to the complex. However, S5a was not found in the free state in HeLa cells. Besides, all subunits in PA700, including S5a, exchanged at similar low rates. It therefore seems that 26-S proteasomes function as stable entities during...

  3. Proteins interacting with the 26S proteasome

    DEFF Research Database (Denmark)

    Hartmann-Petersen, R; Gordon, C

    2004-01-01

    The 26S proteasome is the multi-protein protease that recognizes and degrades ubiquitinylated substrates targeted for destruction by the ubiquitin pathway. In addition to the well-documented subunit organization of the 26S holoenzyme, it is clear that a number of other proteins transiently...... associate with the 26S complex. These transiently associated proteins confer a number of different roles such as substrate presentation, cleavage of the multi-ubiquitin chain from the protein substrate and turnover of misfolded proteins. Such activities are essential for the 26S proteasome to efficiently...... fulfill its intracellular function in protein degradation....

  4. Structural analysis of the 26S proteasome by cryoelectron tomography

    International Nuclear Information System (INIS)

    Nickell, Stephan; Mihalache, Oana; Beck, Florian; Hegerl, Reiner; Korinek, Andreas; Baumeister, Wolfgang

    2007-01-01

    The 26S proteasome is the key enzyme of intracellular protein degradation in eukaryotic cells. It is a multisubunit complex of 2.5 MDa confining the proteolytic action to an inner compartment with tightly controlled access. Structural studies of this intriguing molecular machine have been hampered by its intrinsic instability and its dynamics. Here we have used an unconventional approach to obtain a three-dimensional structure of the holocomplex uncompromised by preparation-induced alterations and unbiased by any starting model. We have performed a tomographic reconstruction, followed by averaging over approx. 150 individual reconstructions, of Drosophila 26S proteasomes suspended in a thin layer of amorphous ice

  5. Molecular characterization of 26S proteasome regulatory subunit in ...

    African Journals Online (AJOL)

    Trichophyton verrucosum is a zoophilic dermatophyte, which causes dermatophytosis infection in human as well as animals. 26S proteasome is an important protein in eukaryotic cells that is involved with degradation of unneeded or damaged proteins, when tagged with ubiquitin. In this study, we characterized the 26S ...

  6. Early cysteine-dependent inactivation of 26S proteasomes does not involve particle disassembly

    Directory of Open Access Journals (Sweden)

    Martín Hugo

    2018-06-01

    Full Text Available Under oxidative stress 26S proteasomes suffer reversible disassembly into its 20S and 19S subunits, a process mediated by HSP70. This inhibits the degradation of polyubiquitinated proteins by the 26S proteasome and allows the degradation of oxidized proteins by a free 20S proteasome. Low fluxes of antimycin A-stimulated ROS production caused dimerization of mitochondrial peroxiredoxin 3 and cytosolic peroxiredoxin 2, but not peroxiredoxin overoxidation and overall oxidation of cellular protein thiols. This moderate redox imbalance was sufficient to inhibit the ATP stimulation of 26S proteasome activity. This process was dependent on reversible cysteine oxidation. Moreover, our results show that this early inhibition of ATP stimulation occurs previous to particle disassembly, indicating an intermediate step during the redox regulation of the 26S proteasome with special relevance under redox signaling rather than oxidative stress conditions.

  7. Nucleolar Proteome Analysis and Proteasomal Activity Assays Reveal a Link between Nucleolus and 26S Proteasome in A. thaliana

    Science.gov (United States)

    Montacié, Charlotte; Durut, Nathalie; Opsomer, Alison; Palm, Denise; Comella, Pascale; Picart, Claire; Carpentier, Marie-Christine; Pontvianne, Frederic; Carapito, Christine; Schleiff, Enrico; Sáez-Vásquez, Julio

    2017-01-01

    In all eukaryotic cells, the nucleolus is functionally and structurally linked to rRNA synthesis and ribosome biogenesis. This compartment contains as well factors involved in other cellular activities, but the functional interconnection between non-ribosomal activities and the nucleolus (structure and function) still remains an open question. Here, we report a novel mass spectrometry analysis of isolated nucleoli from Arabidopsis thaliana plants using the FANoS (Fluorescence Assisted Nucleolus Sorting) strategy. We identified many ribosome biogenesis factors (RBF) and proteins non-related with ribosome biogenesis, in agreement with the recognized multi-functionality of the nucleolus. Interestingly, we found that 26S proteasome subunits localize in the nucleolus and demonstrated that proteasome activity and nucleolus organization are intimately linked to each other. Proteasome subunits form discrete foci in the disorganized nucleolus of nuc1.2 plants. Nuc1.2 protein extracts display reduced proteasome activity in vitro compared to WT protein extracts. Remarkably, proteasome activity in nuc1.2 is similar to proteasome activity in WT plants treated with proteasome inhibitors (MG132 or ALLN). Finally, we show that MG132 treatment induces disruption of nucleolar structures in WT but not in nuc1.2 plants. Altogether, our data suggest a functional interconnection between nucleolus structure and proteasome activity. PMID:29104584

  8. Txl1 and Txc1 are co-factors of the 26S proteasome in fission yeast

    DEFF Research Database (Denmark)

    Andersen, Katrine M; Jensen, Camilla; Kriegenburg, Franziska

    2011-01-01

    and thereby equips proteasomes with redox capabilities. Here, we characterize the fission yeast orthologue of Txnl1, called Txl1. Txl1 associates with the 26S proteasome via its C-terminal domain. This domain is also found in the uncharacterized protein, Txc1, which was also found to interact with 26S...

  9. Poly-Ub-substrate-degradative activity of 26S proteasome is not impaired in the aging rat brain.

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    Carolin Giannini

    Full Text Available Proteostasis is critical for the maintenance of life. In neuronal cells an imbalance between protein synthesis and degradation is thought to be involved in the pathogenesis of neurodegenerative diseases during aging. Partly, this seems to be due to a decrease in the activity of the ubiquitin-proteasome system, wherein the 20S/26S proteasome complexes catalyse the proteolytic step. We have characterised 20S and 26S proteasomes from cerebrum, cerebellum and hippocampus of 3 weeks old (young and 24 month old (aged rats. Our data reveal that the absolute amount of the proteasome is not dfferent between both age groups. Within the majority of standard proteasomes in brain the minute amounts of immuno-subunits are slightly increased in aged rat brain. While this goes along with a decrease in the activities of 20S and 26S proteasomes to hydrolyse synthetic fluorogenic tripeptide substrates from young to aged rats, the capacity of 26S proteasomes for degradation of poly-Ub-model substrates and its activation by poly-Ub-substrates is not impaired or even slightly increased in brain of aged rats. We conclude that these alterations in proteasome properties are important for maintaining proteostasis in the brain during an uncomplicated aging process.

  10. Thioredoxin Txnl1/TRP32 Is a Redox-active Cofactor of the 26 S Proteasome

    DEFF Research Database (Denmark)

    Andersen, Katrine M; Klausen, Louise Kjær; Prag, Søren

    2009-01-01

    in the cytoplasm and nucleus. Txnl1 has thioredoxin activity with a redox potential of about -250 mV. Mutant Txnl1 with one active site cysteine replaced by serine formed disulfide bonds to eEF1A1, a substrate-recruiting factor of the 26S proteasome. eEF1A1 is therefore a likely physiological substrate....... In response to knock-down of Txnl1, ubiquitin-protein conjugates were moderately stabilised. Hence, Txnl1 is the first example of a direct connection between protein reduction and proteolysis, two major intracellular protein quality control mechanisms....

  11. Compensatory role of the Nrf2–ARE pathway against paraquat toxicity: Relevance of 26S proteasome activity

    Directory of Open Access Journals (Sweden)

    Yasuhiko Izumi

    2015-11-01

    Full Text Available Oxidative stress and the ubiquitin–proteasome system play a key role in the pathogenesis of Parkinson disease. Although the herbicide paraquat is an environmental factor that is involved in the etiology of Parkinson disease, the role of 26S proteasome in paraquat toxicity remains to be determined. Using PC12 cells overexpressing a fluorescent protein fused to the proteasome degradation signal, we report here that paraquat yielded an inhibitory effect on 26S proteasome activity without an obvious decline in 20S proteasome activity. Relative low concentrations of proteasome inhibitors caused the accumulation of nuclear factor erythroid 2-related factor 2 (Nrf2, which is targeted to the ubiquitin–proteasome system, and activated the antioxidant response element (ARE-dependent transcription. Paraquat also upregulated the protein level of Nrf2 without increased expression of Nrf2 mRNA, and activated the Nrf2–ARE pathway. Consequently, paraquat induced expression of Nrf2-dependent ARE-driven genes, such as γ-glutamylcysteine synthetase, catalase, and hemeoxygenase-1. Knockdown of Nrf2 or inhibition of γ-glutamylcysteine synthetase and catalase exacerbated paraquat-induced toxicity, whereas suppression of hemeoxygenase-1 did not. These data indicate that the compensatory activation of the Nrf2–ARE pathway via inhibition of 26S proteasome serves as part of a cellular defense mechanism to protect against paraquat toxicity.

  12. Proteomics of the 26S proteasome in Spodoptera frugiperda cells infected with the nucleopolyhedrovirus, AcMNPV.

    Science.gov (United States)

    Lyupina, Yulia V; Zatsepina, Olga G; Serebryakova, Marina V; Erokhov, Pavel A; Abaturova, Svetlana B; Kravchuk, Oksana I; Orlova, Olga V; Beljelarskaya, Svetlana N; Lavrov, Andrey I; Sokolova, Olga S; Mikhailov, Victor S

    2016-06-01

    Baculoviruses are large DNA viruses that infect insect species such as Lepidoptera and are used in biotechnology for protein production and in agriculture as insecticides against crop pests. Baculoviruses require activity of host proteasomes for efficient reproduction, but how they control the cellular proteome and interact with the ubiquitin proteasome system (UPS) of infected cells remains unknown. In this report, we analyzed possible changes in the subunit composition of 26S proteasomes of the fall armyworm, Spodoptera frugiperda (Sf9), cells in the course of infection with the Autographa californica multiple nucleopolyhedrovirus (AcMNPV). 26S proteasomes were purified from Sf9 cells by an immune affinity method and subjected to 2D gel electrophoresis followed by MALDI-TOF mass spectrometry and Mascot search in bioinformatics databases. A total of 34 homologues of 26S proteasome subunits of eukaryotic species were identified including 14 subunits of the 20S core particle (7 α and 7 β subunits) and 20 subunits of the 19S regulatory particle (RP). The RP contained homologues of 11 of RPN-type and 6 of RPT-type subunits, 2 deubiquitinating enzymes (UCH-14/UBP6 and UCH-L5/UCH37), and thioredoxin. Similar 2D-gel maps of 26S proteasomes purified from uninfected and AcMNPV-infected cells at 48hpi confirmed the structural integrity of the 26S proteasome in insect cells during baculovirus infection. However, subtle changes in minor forms of some proteasome subunits were detected. A portion of the α5(zeta) cellular pool that presumably was not associated with the proteasome underwent partial proteolysis at a late stage in infection. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Protection against murine osteoarthritis by inhibition of the 26S proteasome and lysine-48 linked ubiquitination.

    Science.gov (United States)

    Radwan, Marta; Wilkinson, David J; Hui, Wang; Destrument, Auriane P M; Charlton, Sarah H; Barter, Matt J; Gibson, Beth; Coulombe, Josée; Gray, Douglas A; Rowan, Andrew D; Young, David A

    2015-08-01

    To determine whether the process of ubiquitination and/or activity of the 26S proteasome are involved in the induction of osteoarthritis (OA). Bovine cartilage resorption assays, chondrocyte cell-line SW1353 and primary human articular chondrocytes were used with the general proteasome inhibitor MG132 or vehicle to identify a role of the ubiquitin-proteasome system (UPS) in cartilage destruction and matrix metalloproteinase-13 (MMP13) expression. In vivo, MG132 or vehicle, were delivered subcutaneously to mice following destabilisation of the medial meniscus (DMM)-induced OA. Subsequently, DMM was induced in Lys-to-Arg (K48R and K63R) mutant ubiquitin (Ub) transgenic mice. Cytokine signalling in SW1353s was monitored by immunoblotting and novel ubiquitinated substrates identified using Tandem Ubiquitin Binding Entities purification followed by mass spectrometry. The ubiquitination of TRAFD1 was assessed via immunoprecipitation and immunoblotting and its role in cytokine signal-transduction determined using RNA interference and real-time RT-PCR for MMP13 and interleukin-6 (IL6). Supplementation with the proteasome inhibitor MG132 protected cartilage from cytokine-mediated resorption and degradation in vivo in mice following DMM-induced OA. Using transgenic animals only K48R-mutated Ub partially protected against OA compared to wild-type or wild-type Ub transgenic mice, and this was only evident on the medial femoral condyle. After confirming ubiquitination was vital for NF-κB signalling and MMP13 expression, a screen for novel ubiquitinated substrates involved in cytokine-signalling identified TRAFD1; the depletion of which reduced inflammatory mediator-induced MMP13 and IL6 expression. Our data for the first time identifies a role for ubiquitination and the proteasome in the induction of OA via regulation of inflammatory mediator-induced MMP13 expression. These data open avenues of research to determine whether the proteasome, or K48-linked ubiquitination, are

  14. Uch2/Uch37 is the major deubiquitinating enzyme associated with the 26S proteasome in fission yeast

    DEFF Research Database (Denmark)

    Stone, Miranda; Hartmann-Petersen, Rasmus; Seeger, Michael

    2004-01-01

    . Some deubiquitinating enzymes are associated with the 26S proteasome contributing to and regulating the particle's activity. Here, we characterise fission yeast Uch2 and Ubp6, two proteasome associated deubiquitinating enzymes. The human orthologues of these enzymes are known as Uch37 and Usp14......, respectively. We report that the subunit Uch2/Uch37 is the major deubiquitinating enzyme associated with the fission yeast 26S proteasome. In contrast, the activity of Ubp6 appears to play a more regulatory and/or structural role involving the proteasome subunits Mts1/Rpn9, Mts2/Rpt2 and Mts3/Rpn12, as Ubp6...... becomes essential when activity of these subunits is compromised by conditional mutations. Finally, when the genes encoding Uch2/Uch37 and Ubp6 are disrupted, the cells are viable without showing obvious signs of impaired ubiquitin-dependent proteolysis, indicating that other deubiquitinating enzymes may...

  15. Basic leucine zipper protein Cnc-C is a substrate and transcriptional regulator of the Drosophila 26S proteasome.

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    Grimberg, Kristian Björk; Beskow, Anne; Lundin, Daniel; Davis, Monica M; Young, Patrick

    2011-02-01

    While the 26S proteasome is a key proteolytic complex, little is known about how proteasome levels are maintained in higher eukaryotic cells. Here we describe an RNA interference (RNAi) screen of Drosophila melanogaster that was used to identify transcription factors that may play a role in maintaining levels of the 26S proteasome. We used an RNAi library against 993 Drosophila transcription factor genes to identify genes whose suppression in Schneider 2 cells stabilized a ubiquitin-green fluorescent protein reporter protein. This screen identified Cnc (cap 'n' collar [CNC]; basic region leucine zipper) as a candidate transcriptional regulator of proteasome component expression. In fact, 20S proteasome activity was reduced in cells depleted of cnc. Immunoblot assays against proteasome components revealed a general decline in both 19S regulatory complex and 20S proteasome subunits after RNAi depletion of this transcription factor. Transcript-specific silencing revealed that the longest of the seven transcripts for the cnc gene, cnc-C, was needed for proteasome and p97 ATPase production. Quantitative reverse transcription-PCR confirmed the role of Cnc-C in activation of transcription of genes encoding proteasome components. Expression of a V5-His-tagged form of Cnc-C revealed that the transcription factor is itself a proteasome substrate that is stabilized when the proteasome is inhibited. We propose that this single cnc gene in Drosophila resembles the ancestral gene family of mammalian nuclear factor erythroid-derived 2-related transcription factors, which are essential in regulating oxidative stress and proteolysis.

  16. Mouse homologue of yeast Prp19 interacts with mouse SUG1, the regulatory subunit of 26S proteasome

    International Nuclear Information System (INIS)

    Sihn, Choong-Ryoul; Cho, Si Young; Lee, Jeong Ho; Lee, Tae Ryong; Kim, Sang Hoon

    2007-01-01

    Yeast Prp19 has been shown to involve in pre-mRNA splicing and DNA repair as well as being an ubiquitin ligase. Mammalian homologue of yeast Prp19 also plays on similar functional activities in cells. In the present study, we isolated mouse SUG1 (mSUG1) as binding partner of mouse Prp19 (mPrp19) by the yeast two-hybrid system. We confirmed the interaction of mPrp9 with mSUG1 by GST pull-down assay and co-immunoprecipitation assay. The N-terminus of mPrp19 including U-box domain was associated with the C-terminus of mSUG1. Although, mSUG1 is a regulatory subunit of 26S proteasome, mPrp19 was not degraded in the proteasome-dependent pathway. Interestingly, GFP-mPrp19 fusion protein was co-localized with mSUG1 protein in cytoplasm as the formation of the speckle-like structures in the presence of a proteasome inhibitor MG132. In addition, the activity of proteasome was increased in cells transfected with mPrp19. Taken together, these results suggest that mPrp19 involves the regulation of protein turnover and may transport its substrates to 26S proteasome through mSUG1 protein

  17. Quaternary structure of the ATPase complex of human 26S proteasomes determined by chemical cross-linking

    DEFF Research Database (Denmark)

    Hartmann-Petersen, R; Tanaka, K; Hendil, K B

    2001-01-01

    and substrate specificity. Among the approximately 18 subunits of PA700 regulator, six are ATPases. The ATPases presumably recognize, unfold, and translocate substrates into the interior of the 26S proteasome. It is generally believed that the ATPases form a hexameric ring. By means of chemical cross......-linking, immunoprecipitation, and blotting, we have determined that the ATPases are organized in the order S6-S6'-S10b-S8-S4-S7. Additionally, we found cross-links between the ATPase S10b and the 20S proteasome subunit alpha6. Together with the previously known interaction between S8 and alpha1 and between S4 and alpha7......, these data establish the relative orientations of ATPases with respect to the 20S proteasome....

  18. Inhibition of the 26S proteasome blocks progesterone receptor-dependent transcription through failed recruitment of RNA polymerase II.

    Science.gov (United States)

    Dennis, Andrew P; Lonard, David M; Nawaz, Zafar; O'Malley, Bert W

    2005-03-01

    In the present study, we investigated the involvement of protein degradation via the 26S proteasome during progesterone receptor (PR)-mediated transcription in T-47D cells containing a stably integrated MMTV-CAT reporter construct (CAT0 cells). Progesterone induced CAT and HSD11beta2 transcription while co-treatment with the proteasome inhibitor, MG132, blocked PR-induced transcription in a time-dependent fashion. MG132 treatment also inhibited transcription of beta-actin and cyclophilin, but not two proteasome subunit genes, PSMA1 and PSMC1, indicating that proteasome inhibition affects a subset of RNA polymerase II (RNAP(II))-regulated genes. Progesterone-mediated recruitment of RNAP(II) was blocked by MG132 treatment at time points later than 1 h that was not dependent on the continued presence of PR, associated cofactors, and components of the general transcription machinery, supporting the concept that proteasome-mediated degradation is needed for continued transcription. Surprisingly, progesterone-mediated acetylation of histone H4 was inhibited by MG132 with the concomitant recruitment of HDAC3, NCoR, and SMRT. We demonstrate that the steady-state protein levels of SMRT and NCoR are higher in the presence of MG132 in CAT0 cells, consistent with other reports that SMRT and NCoR are targets of the 26S proteasome. However, inhibition of histone deacetylation by trichostatin A (TSA) treatment or SMRT/NCoR knockdown by siRNA did not restore MG132-inhibited progesterone-dependent transcription. Therefore, events other than histone deacetylation and stability of SMRT and NCoR must also play a role in inhibition of PR-mediated transcription.

  19. HIV-1 replication through hHR23A-mediated interaction of Vpr with 26S proteasome.

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    Ge Li

    2010-06-01

    Full Text Available HIV-1 Vpr is a virion-associated protein. Its activities link to viral pathogenesis and disease progression of HIV-infected patients. In vitro, Vpr moderately activates HIV-1 replication in proliferating T cells, but it is required for efficient viral infection and replication in vivo in non-dividing cells such as macrophages. How exactly Vpr contributes to viral replication remains elusive. We show here that Vpr stimulates HIV-1 replication at least in part through its interaction with hHR23A, a protein that binds to 19S subunit of the 26S proteasome and shuttles ubiquitinated proteins to the proteasome for degradation. The Vpr-proteasome interaction was initially discovered in fission yeast, where Vpr was shown to associate with Mts4 and Mts2, two 19S-associated proteins. The interaction of Vpr with the 19S subunit of the proteasome was further confirmed in mammalian cells where Vpr associates with the mammalian orthologues of fission yeast Mts4 and S5a. Consistently, depletion of hHR23A interrupts interaction of Vpr with proteasome in mammalian cells. Furthermore, Vpr promotes hHR23A-mediated protein-ubiquitination, and down-regulation of hHR23A using RNAi significantly reduced viral replication in non-proliferating MAGI-CCR5 cells and primary macrophages. These findings suggest that Vpr-proteasome interaction might counteract certain host restriction factor(s to stimulate viral replication in non-dividing cells.

  20. Plant WEE1 kinase is cell cycle regulated and removed at mitosis via the 26S proteasome machinery

    Science.gov (United States)

    Cook, Gemma S.; Grønlund, Anne Lentz; Siciliano, Ilario; Spadafora, Natasha; Amini, Maryam; Herbert, Robert J.; Bitonti, M. Beatrice; Graumann, Katja; Francis, Dennis; Rogers, Hilary J.

    2013-01-01

    In yeasts and animals, premature entry into mitosis is prevented by the inhibitory phosphorylation of cyclin-dependent kinase (CDK) by WEE1 kinase, and, at mitosis, WEE1 protein is removed through the action of the 26S proteasome. Although in higher plants WEE1 function has been confirmed in the DNA replication checkpoint, Arabidopsis wee1 insertion mutants grow normally, and a role for the protein in the G2/M transition during an unperturbed plant cell cycle is yet to be confirmed. Here data are presented showing that the inhibitory effect of WEE1 on CDK activity in tobacco BY-2 cell cultures is cell cycle regulated independently of the DNA replication checkpoint: it is high during S-phase but drops as cells traverse G2 and enter mitosis. To investigate this mechanism further, a yeast two-hybrid screen was undertaken to identify proteins interacting with Arabidopsis WEE1. Three F-box proteins and a subunit of the proteasome complex were identified, and bimolecular fluorescence complementation confirmed an interaction between AtWEE1 and the F-box protein SKP1 INTERACTING PARTNER 1 (SKIP1). Furthermore, the AtWEE1–green fluorescent protein (GFP) signal in Arabidopsis primary roots treated with the proteasome inhibitor MG132 was significantly increased compared with mock-treated controls. Expression of AtWEE1–YFPC (C-terminal portion of yellow fluorescent protein) or AtWEE1 per se in tobacco BY-2 cells resulted in a premature increase in the mitotic index compared with controls, whereas co-expression of AtSKIP1–YFPN negated this effect. These data support a role for WEE1 in a normal plant cell cycle and its removal at mitosis via the 26S proteasome. PMID:23536609

  1. 26S Proteasome regulation of Ankrd1/CARP in adult rat ventricular myocytes and human microvascular endothelial cells

    International Nuclear Information System (INIS)

    Samaras, Susan E.; Chen, Billy; Koch, Stephen R.; Sawyer, Douglas B.; Lim, Chee Chew; Davidson, Jeffrey M.

    2012-01-01

    Highlights: ► The 26S proteasome regulates Ankrd1 levels in cardiomyocytes and endothelial cells. ► Ankrd1 protein degrades 60-fold faster in endothelial cells than cardiomyocytes. ► Differential degradation appears related to nuclear vs. sarcolemmal localization. ► Endothelial cell density shows uncoupling of Ankrd1 mRNA and protein levels. -- Abstract: Ankyrin repeat domain 1 protein (Ankrd1), also known as cardiac ankyrin repeat protein (CARP), increases dramatically after tissue injury, and its overexpression improves aspects of wound healing. Reports that Ankrd1/CARP protein stability may affect cardiovascular organization, together with our findings that the protein is crucial to stability of the cardiomyocyte sarcomere and increased in wound healing, led us to compare the contribution of Ankrd1/CARP stability to its abundance. We found that the 26S proteasome is the dominant regulator of Ankrd1/CARP degradation, and that Ankrd1/CARP half-life is significantly longer in cardiomyocytes (h) than endothelial cells (min). In addition, higher endothelial cell density decreased the abundance of the protein without affecting steady state mRNA levels. Taken together, our data and that of others indicate that Ankrd1/CARP is highly regulated at multiple levels of its expression. The striking difference in protein half-life between a muscle and a non-muscle cell type suggests that post-translational proteolysis is correlated with the predominantly structural versus regulatory role of the protein in the two cell types.

  2. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants

    OpenAIRE

    Baudisch, Bianca; Pfort, Ingrid; Sorge, Eberhard; Conrad, Udo

    2018-01-01

    Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  3. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants

    Directory of Open Access Journals (Sweden)

    Bianca Baudisch

    2018-02-01

    Full Text Available Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  4. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants.

    Science.gov (United States)

    Baudisch, Bianca; Pfort, Ingrid; Sorge, Eberhard; Conrad, Udo

    2018-01-01

    Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  5. 26S proteasome and insulin-like growth factor-1 in serum of dogs suffering from malignant tumors.

    Science.gov (United States)

    Gerke, Ingrid; Kaup, Franz-Josef; Neumann, Stephan

    2018-04-01

    Studies in humans have shown that the ubiquitin-proteasome pathway and the insulin-like growth factor axis are involved in carcinogenesis, thus, components of these systems might be useful as prognostic markers and constitute potential therapeutic targets. In veterinary medicine, only a few studies exist on this topic. Here, serum concentrations of 26S proteasome (26SP) and insulin-like growth factor-1 (IGF-1) were measured by canine enzyme-linked immunosorbent assay (ELISA) in 43 dogs suffering from malignant tumors and 21 clinically normal dogs (control group). Relationships with tumor size, survival time, body condition score (BCS), and tumor entity were assessed. The median 26SP concentration in the tumor group was non-significantly higher than in the control group. However, dogs with mammary carcinomas displayed significantly increased 26SP levels compared to the control group and dogs with tumor size less than 5 cm showed significantly increased 26SP concentrations compared to dogs with larger tumors and control dogs. 26SP concentrations were not correlated to survival time or BCS. No significant difference in IGF-1 levels was found between the tumor group and the control group; however, IGF-1 concentrations displayed a larger range of values in the tumor group. Dogs with tumors greater than 5 cm showed significantly higher IGF-1 levels than dogs with smaller tumors. The IGF-1 concentrations were positively correlated to survival time, but no correlation with BCS was found. Consequently, serum 26SP concentrations seem to be increased in some dogs suffering from malignant tumors, especially in dogs with mammary carcinoma and smaller tumors. Increased serum IGF-1 concentrations could be an indication of large tumors and a poor prognosis.

  6. Dietary apigenin potentiates the inhibitory effect of interferon-α on cancer cell viability through inhibition of 26S proteasome-mediated interferon receptor degradation

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    Sheng Li

    2016-06-01

    Full Text Available Background: Type I interferons (IFN-α/β have broad and potent immunoregulatory and antiproliferative activities. However, it is still known whether the dietary flavonoids exhibit their antiviral and anticancer properties by modulating the function of type I IFNs. Objective: This study aimed at determining the role of apigenin, a dietary plant flavonoid abundant in common fruits and vegetables, on the type I IFN-mediated inhibition of cancer cell viability. Design: Inhibitory effect of apigenin on human 26S proteasome, a known negative regulator of type I IFN signaling, was evaluated in vitro. Molecular docking was conducted to know the interaction between apigenin and subunits of 26S proteasome. Effects of apigenin on JAK/STAT pathway, 26S proteasome-mediated interferon receptor stability, and cancer cells viability were also investigated. Results: Apigenin was identified to be a potent inhibitor of human 26S proteasome in a cell-based assay. Apigenin inhibited the chymotrypsin-like, caspase-like, and trypsin-like activities of the human 26S proteasome and increased the ubiquitination of endogenous proteins in cells. Results from computational modeling of the potential interactions of apigenin with the chymotrypsin site (β5 subunit, caspase site (β1 subunit, and trypsin site (β2 subunit of the proteasome were consistent with the observed proteasome inhibitory activity. Apigenin enhanced the phosphorylation of signal transducer and activator of transcription proteins (STAT1 and STAT2 and promoted the endogenous IFN-α-regulated gene expression. Apigenin inhibited the IFN-α-stimulated ubiquitination and degradation of type I interferon receptor 1 (IFNAR1. Apigenin also sensitized the inhibitory effect of IFN-α on viability of cervical carcinoma HeLa cells. Conclusion: These results suggest that apigenin potentiates the inhibitory effect of IFN-α on cancer cell viability by activating JAK/STAT signaling pathway through inhibition of 26S

  7. An AAA Motor-Driven Mechanical Switch in Rpn11 Controls Deubiquitination at the 26S Proteasome.

    Science.gov (United States)

    Worden, Evan J; Dong, Ken C; Martin, Andreas

    2017-09-07

    Poly-ubiquitin chains direct protein substrates to the 26S proteasome, where they are removed by the deubiquitinase Rpn11 during ATP-dependent substrate degradation. Rapid deubiquitination is required for efficient degradation but must be restricted to committed substrates that are engaged with the ATPase motor to prevent premature ubiquitin chain removal and substrate escape. Here we reveal the ubiquitin-bound structure of Rpn11 from S. cerevisiae and the mechanisms for mechanochemical coupling of substrate degradation and deubiquitination. Ubiquitin binding induces a conformational switch of Rpn11's Insert-1 loop from an inactive closed state to an active β hairpin. This switch is rate-limiting for deubiquitination and strongly accelerated by mechanical substrate translocation into the AAA+ motor. Deubiquitination by Rpn11 and ubiquitin unfolding by the ATPases are in direct competition. The AAA+ motor-driven acceleration of Rpn11 is therefore important to ensure that poly-ubiquitin chains are removed only from committed substrates and fast enough to prevent their co-degradation. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Induction of 26S proteasome subunit PSMB5 by the bifunctional inducer 3-methylcholanthrene through the Nrf2-ARE, but not the AhR/Arnt-XRE, pathway

    International Nuclear Information System (INIS)

    Kwak, Mi-Kyoung; Kensler, Thomas W.

    2006-01-01

    The 26S proteasome is responsible for degradation of abnormal intracellular proteins, including oxidatively damaged proteins and may play a role as a component of a cellular antioxidative system. However, little is known about regulation of proteasome expression. In the present study, regulation of proteasome expression by the bifunctional enzyme inducer and a specific signaling pathway for this regulation were investigated in murine neuroblastoma cells. Expression of catalytic core subunits including PSMB5 and peptidase activities of the proteasome were elevated following incubation with 3-methylcholanthrene (3-MC). Studies using reporter genes containing the murine Psmb5 promoter showed that transcriptional activity of this gene was enhanced by 3-MC. Overexpression of AhR/Arnt did not affect activation of the Pmsb5 promoter by 3-MC and deletion of the xenobiotic response elements (XREs) from this promoter exerted modest effects on inducibility in response to 3-MC. However, mutation of the proximal AREs of the Psmb5 promoter largely abrogated its inducibility by 3-MC. In addition, this promoter showed a blunted response toward 3-MC in the absence of nrf2; 3-MC incubation increased nuclear levels of Nrf2 only in wild-type cells. Collectively, these results indicate that expression of proteasome subunit PSMB5 is modulated by bifunctional enzyme inducers in a manner independent of the AhR/Arnt-XRE pathway but dependent upon the Nrf2-ARE pathway

  9. SAMe prevents the induction of the immunoproteasome and preserves the 26S proteasome in the DDC-induced MDB mouse model.

    Science.gov (United States)

    Bardag-Gorce, Fawzia; Oliva, Joan; Li, Jun; French, Barbara A; French, Samuel W

    2010-06-01

    Mallory-Denk bodies (MDBs) form in the liver of alcoholic patients. This occurs because of the accumulation and aggregation of ubiquitinated cytokeratins, which hypothetically is due to the ubiquitin-proteasome pathway's (UPP) failure to degrade the cytokeratins. The experimental model of MDB formation was used in which MDBs were induced by refeeding DDC to drug-primed mice. The gene expression and protein levels of LMP2, LMP7 and MECL-1, the catalytic subunits in the immunoproteasome, as well as FAT10, were increased in the liver cells forming MDBs but not in the intervening normal hepatocytes. Chymotrypsin-like activity of the UPP was decreased by DDC refeeding, indicating that a switch from the UPP to the immunoproteasome had occurred at the expense of the 26S proteasome. The failure of the UPP to digest cytokeratins would explain MDB aggregate formation. SAMe prevented the decrease in UPP activity, the increase in LMP2, LMP7, and MECL-1 protein levels and MDB formation induced by DDC. DDC refeeding also induced the TNFalpha and IFNgamma receptors. SAMe prevented the increase in the TNFalpha and IFNgamma receptors, supporting the idea that TNFalpha and IFNgamma were responsible for the up regulation of LMP2, LPM7, and FAT10. These results support the conclusion that MDBs form in FAT10 over-expressing hepatocytes where the up regulation of the immunoproteasome occurs at the expense of the 26S proteasome. Copyright 2010 Elsevier Inc. All rights reserved.

  10. The 26S Proteasome Degrades the Soluble but Not the Fibrillar Form of the Yeast Prion Ure2p In Vitro.

    Directory of Open Access Journals (Sweden)

    Kai Wang

    Full Text Available Yeast prions are self-perpetuating protein aggregates that cause heritable and transmissible phenotypic traits. Among these, [PSI+] and [URE3] stand out as the most studied yeast prions, and result from the self-assembly of the translation terminator Sup35p and the nitrogen catabolism regulator Ure2p, respectively, into insoluble fibrillar aggregates. Protein quality control systems are well known to govern the formation, propagation and transmission of these prions. However, little is known about the implication of the cellular proteolytic machineries in their turnover. We previously showed that the 26S proteasome degrades both the soluble and fibrillar forms of Sup35p and affects [PSI+] propagation. Here, we show that soluble native Ure2p is degraded by the proteasome in an ubiquitin-independent manner. Proteasomal degradation of Ure2p yields amyloidogenic N-terminal peptides and a C-terminal resistant fragment. In contrast to Sup35p, fibrillar Ure2p resists proteasomal degradation. Thus, structural variability within prions may dictate their ability to be degraded by the cellular proteolytic systems.

  11. The 26S Proteasome Degrades the Soluble but Not the Fibrillar Form of the Yeast Prion Ure2p In Vitro

    Science.gov (United States)

    Wang, Kai; Redeker, Virginie; Madiona, Karine; Melki, Ronald; Kabani, Mehdi

    2015-01-01

    Yeast prions are self-perpetuating protein aggregates that cause heritable and transmissible phenotypic traits. Among these, [PSI +] and [URE3] stand out as the most studied yeast prions, and result from the self-assembly of the translation terminator Sup35p and the nitrogen catabolism regulator Ure2p, respectively, into insoluble fibrillar aggregates. Protein quality control systems are well known to govern the formation, propagation and transmission of these prions. However, little is known about the implication of the cellular proteolytic machineries in their turnover. We previously showed that the 26S proteasome degrades both the soluble and fibrillar forms of Sup35p and affects [PSI +] propagation. Here, we show that soluble native Ure2p is degraded by the proteasome in an ubiquitin-independent manner. Proteasomal degradation of Ure2p yields amyloidogenic N-terminal peptides and a C-terminal resistant fragment. In contrast to Sup35p, fibrillar Ure2p resists proteasomal degradation. Thus, structural variability within prions may dictate their ability to be degraded by the cellular proteolytic systems. PMID:26115123

  12. The devil is in the details: comparison between COP9 signalosome (CSN) and the LID of the 26S proteasome.

    Science.gov (United States)

    Meister, Cindy; Gulko, Miriam Kolog; Köhler, Anna M; Braus, Gerhard H

    2016-02-01

    The COP9 signalosome (CSN) and the proteasomal LID are conserved macromolecular complexes composed of at least eight subunits with molecular weights of approximately 350 kDa. CSN and LID are part of the ubiquitin–proteasome pathway and cleave isopeptide linkages of lysine side chains on target proteins. CSN cleaves the isopeptide bond of ubiquitin-like protein Nedd8 from cullins, whereas the LID cleaves ubiquitin from target proteins sentenced for degradation. CSN and LID are structurally and functionally similar but the order of the assembly pathway seems to be different. The assembly differs in at least the last subunit joining the pre-assembled subcomplex. This review addresses the similarities and differences in structure, function and assembly of CSN and LID.

  13. CRM1-mediated nuclear export is required for 26 S proteasome-dependent degradation of the TRIP-Br2 proto-oncoprotein.

    Science.gov (United States)

    Cheong, Jit Kong; Gunaratnam, Lakshman; Hsu, Stephen I-Hong

    2008-04-25

    Overexpression of the proto-oncogene TRIP-Br2 (SERTAD2) has been shown to induce E2F activity and promote tumorigenesis, whereas ablation of TRIP-Br2 arrests cell proliferation. Timely degradation of many cell cycle regulators is fundamental to the maintenance of proper cell cycle progression. Here we report novel mechanism(s) that govern the tight regulation of TRIP-Br2 levels during cell cycle progression. TRIP-Br2 was observed to be a short-lived protein in which the expression level peaks at the G(1)/S boundary. TRIP-Br2 accumulated in cells treated with 26 S proteasome inhibitors. Co-immunoprecipitation studies revealed that TRIP-Br2 forms ubiquitin conjugates. In silico analysis identified a putative leucine-rich nuclear export signal (NES) motif that overlaps with the PHD-Bromo interaction domain in the acidic C-terminal transactivation domain (TAD) of TRIP-Br2. This NES motif is highly conserved in widely divergent species and in all TRIP-Br family members. TRIP-Br2 was shown to be stabilized in G(2)/M phase cells through nuclear entrapment, either by deletion of the acidic C-terminal TAD, which includes the NES motif, or by leptomycin B-mediated inhibition of the CRM1-dependent nuclear export machinery. Mutation of leucine residue 238 of this NES motif abolished the interaction between CRM1 and TRIP-Br2, as well as the nuclear export of TRIP-Br2 and its subsequent 26 S proteasome-dependent degradation. These data suggest that CRM1-mediated nuclear export may be required for the proper execution of ubiquitin-proteasome-dependent degradation of TRIP-Br2.

  14. Degradation of the encephalomyocarditis virus and hepatitis A virus 3C proteases by the ubiquitin/26S proteasome system in vivo

    International Nuclear Information System (INIS)

    Schlax, Peter E.; Zhang Jin; Lewis, Elizabeth; Planchart, Antonio; Lawson, T. Glen

    2007-01-01

    We have isolated stably transfected mouse embryonic fibroblast cell lines that inducibly express either the mature encephalomyocarditis virus (EMCV) or hepatitis A virus (HAV) 3C protease and have used these cells to demonstrate that both proteins are subject to degradation in vivo by the ubiquitin/26S proteasome system. The detection of 3C protease expression in these cells requires inducing conditions and the presence of one of several proteasome inhibitors. Both 3C proteases are incorporated into conjugates with ubiquitin in vivo. HAV 3C protease expression has deleterious effects on cell viability, as determined by observation and counting of cells cultured in the absence or presence of inducing conditions. The EMCV 3C protease was found to be preferentially localized to the nucleus of induced cells, while the HAV 3C protease remains in the cytoplasm. The absence of polyubiquitinated EMCV 3C protease conjugates in nuclear fraction preparations suggests that localization to the nucleus can protect this protein from ubiquitination

  15. Evaluation of serum insulin-like growth factor-1 and 26S proteasome concentrations in healthy dogs and dogs with chronic diseases depending on body condition score.

    Science.gov (United States)

    Gerke, Ingrid; Kaup, Franz-Josef; Neumann, Stephan

    2018-06-01

    In patients suffering from chronic diseases, the objective assessment of metabolic states could be of interest for disease prognosis and therapeutic options. Therefore, the aim of this study was to assess insulin-like growth factor-1 (IGF-1) and 26S proteasome (26SP) in healthy dogs and dogs suffering from chronic diseases depending on their body condition score (BCS) and to examine their potential for objective assessment of anabolic and catabolic states. Serum concentrations of IGF-1, an anabolic hormone, and 26SP, a multiprotein complex which is part of the ubiquitin-proteasome pathway, by which the majority of endogenous proteins including the muscle proteins are degraded, were measured in 21 healthy dogs and 20 dogs with chronic diseases by canine ELISA. The concentrations of IGF-1, 26SP and their ratio (IGF-1/26SP) were set in relationship to the BCS of the dogs. When examining healthy and chronically diseased dogs separately, a positive correlation between IGF-1 and the BCS was observed in the healthy group and a negative correlation between 26SP and the BCS was noted in dogs with chronic diseases. Further, dogs suffering from chronic diseases showed higher 26SP concentrations and lower values for IGF-1/26SP than the healthy dogs. Overall, we detected a negative correlation between 26SP and the BCS and a positive correlation between IGF-1/26SP and the BCS. The results of our study indicate usability of IGF-1 for description of anabolic states, while 26SP could be useful for detection and description of catabolic states. Finally, the ratio IGF-1/26SP seems to be promising for assessment of metabolic states. Copyright © 2018 Elsevier Ltd. All rights reserved.

  16. Artemisinin disrupts androgen responsiveness of human prostate cancer cells by stimulating the 26S proteasome-mediated degradation of the androgen receptor protein.

    Science.gov (United States)

    Steely, Andrea M; Willoughby, Jamin A; Sundar, Shyam N; Aivaliotis, Vasiliki I; Firestone, Gary L

    2017-10-01

    Androgen receptor (AR) expression and activity is highly linked to the development and progression of prostate cancer and is a target of therapeutic strategies for this disease. We investigated whether the antimalarial drug artemisinin, which is a sesquiterpene lactone isolated from the sweet wormwood plant Artemisia annua, could alter AR expression and responsiveness in cultured human prostate cancer cell lines. Artemisinin treatment induced the 26S proteasome-mediated degradation of the receptor protein, without altering AR transcript levels, in androgen-responsive LNCaP prostate cancer cells or PC-3 prostate cancer cells expressing exogenous wild-type AR. Furthermore, artemisinin stimulated AR ubiquitination and AR receptor interactions with the E3 ubiquitin ligase MDM2 in LNCaP cells. The artemisinin-induced loss of AR protein prevented androgen-responsive cell proliferation and ablated total AR transcriptional activity. The serine/threonine protein kinase AKT-1 was shown to be highly associated with artemisinin-induced proteasome-mediated degradation of AR protein. Artemisinin treatment activated AKT-1 enzymatic activity, enhanced receptor association with AKT-1, and induced AR serine phosphorylation. Treatment of LNCaP cells with the PI3-kinase inhibitor LY294002, which inhibits the PI3-kinase-dependent activation of AKT-1, prevented the artemisinin-induced AR degradation. Furthermore, in transfected receptor-negative PC-3 cells, artemisinin failed to stimulate the degradation of an altered receptor protein (S215A/S792A) with mutations in its two consensus AKT-1 serine phosphorylation sites. Taken together, our results indicate that artemisinin induces the degradation of AR protein and disrupts androgen responsiveness of human prostate cancer cells, suggesting that this natural compound represents a new potential therapeutic molecule that selectively targets AR levels.

  17. Redox-Regulated Pathway of Tyrosine Phosphorylation Underlies NF-κB Induction by an Atypical Pathway Independent of the 26S Proteasome

    Science.gov (United States)

    Cullen, Sarah; Ponnappan, Subramaniam; Ponnappan, Usha

    2015-01-01

    Alternative redox stimuli such as pervanadate or hypoxia/reoxygenation, induce transcription factor NF-κB by phospho-tyrosine-dependent and proteasome-independent mechanisms. While considerable attention has been paid to the absence of proteasomal regulation of tyrosine phosphorylated IκBα, there is a paucity of information regarding proteasomal regulation of signaling events distinct from tyrosine phosphorylation of IκBα. To delineate roles for the ubiquitin-proteasome pathway in the phospho-tyrosine dependent mechanism of NF-κB induction, we employed the proteasome inhibitor, Aclacinomycin, and the phosphotyrosine phosphatase inhibitor, pervanadate (PV). Results from these studies demonstrate that phospho-IκBα (Tyr-42) is not subject to proteasomal degradation in a murine stromal epithelial cell line, confirming results previously reported. Correspondingly, proteasome inhibition had no discernable effect on the key signaling intermediaries, Src and ERK1/2, involved in the phospho-tyrosine mechanisms regulating PV-mediated activation of NF-κB. Consistent with previous reports, a significant redox imbalance leading to the activation of tyrosine kinases, as occurs with pervanadate, is required for the induction of NF-κB. Strikingly, our studies demonstrate that proteasome inhibition can potentiate oxidative stress associated with PV-stimulation without impacting kinase activation, however, other cellular implications for this increase in intracellular oxidation remain to be fully delineated. PMID:25671697

  18. Purification, crystallization and preliminary X-ray diffraction analysis of the non-ATPase subunit Nas6 in complex with the ATPase subunit Rpt3 of the 26S proteasome from Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Nakamura, Yoshihiro; Umehara, Takashi; Tanaka, Akiko; Horikoshi, Masami; Padmanabhan, Balasundaram; Yokoyama, Shigeyuki

    2007-01-01

    The complex of the non-ATPase subunit Nas6 with the C-terminal domain of the ATPase subunit Rpt3 of the 26S proteasome from S. cerevisiae was co-expressed in E. coli and purified to homogeneity. The crystals obtained from the protein complex diffracted to a resolution of 2.2 Å. The non-ATPase subunit Nas6, which is the human orthologue of gankyrin, was co-expressed with the C-terminal domain of the ATPase subunit Rpt3 of the yeast 26S proteasome in Escherichia coli, purified to near-homogeneity and crystallized using the hanging-drop vapour-diffusion method. The protein crystallized in space group P2 1 , with unit-cell parameters a = 60.38, b = 100.22, c = 72.20 Å, β = 94.70° and with three Nas6–Rpt3C molecules per asymmetric unit. The crystal diffracted to beyond 2.2 Å resolution using synchrotron radiation

  19. PUB22 and PUB23 U-BOX E3 ligases directly ubiquitinate RPN6, a 26S proteasome lid subunit, for subsequent degradation in Arabidopsis thaliana

    DEFF Research Database (Denmark)

    Cho, Seok Keun; Bae, Hansol; Ryu, Moonyoung

    2015-01-01

    and PUB23, two U-box E3 ligase homologs, tether ubiquitins to 19S proteasome regulatory particle (RP) subunit RPN6, leading to its degradation. RPN6 was identified as an interacting substrate of PUB22 by yeast two-hybrid screening, and in vitro pull-down assay confirmed that RPN6 interacts not only......Drought stress strongly affects plant growth and development, directly connected with crop yields, accordingly. However, related to the function of U-BOX E3 ligases, the underlying molecular mechanisms of desiccation stress response in plants are still largely unknown. Here we report that PUB22...

  20. Mechanisms promoting and inhibiting the process of proteasomal degradation of cells

    Directory of Open Access Journals (Sweden)

    Pedrycz Agnieszka

    2016-03-01

    Full Text Available Defects in the process of degradation of unneeded cellular proteins underlie many diseases. This article discusses one of the most important systems of removal of abnormal proteins. It describes the process of ubiquitination of proteins for proteasome degradation. It also describes the structure of the 26S and 20S proteasomes and the mechanism of ubiquitin-proteasome system. Proteasome proteolytic system is highly specialized and organized. Protease-proteasome 26S is particularly important for proper cell functioning. It recognizes and degrades marked proteins. Inhibition of proteasome pathway leads to cell cycle arrest and apoptosis.

  1. Regulating the 20S Proteasome Ubiquitin-Independent Degradation Pathway

    Directory of Open Access Journals (Sweden)

    Gili Ben-Nissan

    2014-09-01

    Full Text Available For many years, the ubiquitin-26S proteasome degradation pathway was considered the primary route for proteasomal degradation. However, it is now becoming clear that proteins can also be targeted for degradation by the core 20S proteasome itself. Degradation by the 20S proteasome does not require ubiquitin tagging or the presence of the 19S regulatory particle; rather, it relies on the inherent structural disorder of the protein being degraded. Thus, proteins that contain unstructured regions due to oxidation, mutation, or aging, as well as naturally, intrinsically unfolded proteins, are susceptible to 20S degradation. Unlike the extensive knowledge acquired over the years concerning degradation by the 26S proteasome, relatively little is known about the means by which 20S-mediated proteolysis is controlled. Here, we describe our current understanding of the regulatory mechanisms that coordinate 20S proteasome-mediated degradation, and highlight the gaps in knowledge that remain to be bridged.

  2. Proteasome impairment by α-synuclein.

    Directory of Open Access Journals (Sweden)

    Lisa Zondler

    Full Text Available Parkinson's disease (PD is the second most prevalent neurodegenerative disorder worldwide and characterized by the loss of dopaminergic neurons in the patients' midbrains. Both the presence of the protein α-synuclein in intracellular protein aggregates in surviving neurons and the genetic linking of the α-synuclein encoding gene point towards a major role of α-synuclein in PD etiology. The exact pathogenic mechanisms of PD development are not entirely described to date, neither is the specific role of α-synuclein in this context. Previous studies indicate that one aspect of α-synuclein-related cellular toxicity might be direct proteasome impairment. The 20/26S proteasomal machinery is an important instrument of intracellular protein degradation. Thus, direct proteasome impairment by α-synuclein might explain or at least contribute to the formation of intracellular protein aggregates. Therefore this study investigates direct proteasomal impairment by α-synuclein both in vitro using recombinant α-synuclein and isolated proteasomes as well as in living cells. Our experiments demonstrate that the impairment of proteasome activity by α-synuclein is highly dependent upon the cellular background and origin. We show that recombinant α-synuclein oligomers and fibrils scarcely affect 20S proteasome function in vitro, neither does transient α-synuclein expression in U2OS ps 2042 (Ubi(G76V-GFP cells. However, stable expression of both wild-type and mutant α-synuclein in dopaminergic SH-SY5Y and PC12 cells results in a prominent impairment of the chymotrypsin-like 20S/26S proteasomal protein cleavage. Thus, our results support the idea that α-synuclein in a specific cellular environment, potentially present in dopaminergic cells, cannot be processed by the proteasome and thus contributes to a selective vulnerability of dopaminergic cells to α-synuclein pathology.

  3. Identification and Characterisation of a Proteasome -

    DEFF Research Database (Denmark)

    Andersen, Katrine Mølgaard

    this domain. A txl1 yeast knockout mutant displays a synthetic growth defect with a cut8 knockout, whereas the txc1 knockout does not. In fission yeast, Cut8 is a nuclear protein that tethers the 26S proteasome to the nuclear membrane. In both wild type cells and in a cut8 null mutant Txl1 co......-down of Txnl1 in HeLa cells, whereas no stabilisation was detected in txl1¿ cells in S. pombe. In human cells, an active site Txnl1 mutant formed a mixed disulfide intermediate with eEF1A1, a reported substrate-recruiting factor of the 26S proteasome. Fission yeast Txl1 can reduce an oxidised proteasomal model...

  4. Cytoplasmic proteasomes are not indispensable for cell growth in Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Tsuchiya, Hikaru; Arai, Naoko; Tanaka, Keiji, E-mail: tanaka-kj@igakuken.or.jp; Saeki, Yasushi, E-mail: saeki-ys@igakuken.or.jp

    2013-07-05

    Highlights: •We succeeded to control the proteasome localization by the anchor-away technique. •Nuclear proteasome-depleted cells showed a lethal phenotype. •Cytoplasmic proteasomes are not indispensable for cell growth in dividing cells. -- Abstract: The 26S proteasome is an essential protease complex responsible for the degradation of ubiquitinated proteins in eukaryotic cells. In rapidly proliferating yeast cells, proteasomes are mainly localized in the nucleus, but the biological significance of the proteasome localization is still unclear. In this study, we investigated the relationship between the proteasome localization and the functions by the anchor-away technique, a ligand-dependent sequestration of a target protein into specific compartment(s). Anchoring of the proteasome to the plasma membrane or the ribosome resulted in conditional depletion of the nuclear proteasomes, whereas anchoring to histone resulted in the proteasome sequestration into the nucleus. We observed that the accumulation of ubiquitinated proteins in all the proteasome-targeted cells, suggesting that both the nuclear and cytoplasmic proteasomes have proteolytic functions and that the ubiquitinated proteins are produced and degraded in each compartment. Consistent with previous studies, the nuclear proteasome-depleted cells exhibited a lethal phenotype. In contrast, the nuclear sequestration of the proteasome resulted only in a mild growth defect, suggesting that the cytoplasmic proteasomes are not basically indispensable for cell growth in rapidly growing yeast cells.

  5. Proteasome phosphorylation regulates cocaine-induced sensitization.

    Science.gov (United States)

    Gonzales, Frankie R; Howell, Kristin K; Dozier, Lara E; Anagnostaras, Stephan G; Patrick, Gentry N

    2018-04-01

    Repeated exposure to cocaine produces structural and functional modifications at synapses from neurons in several brain regions including the nucleus accumbens. These changes are thought to underlie cocaine-induced sensitization. The ubiquitin proteasome system plays a crucial role in the remodeling of synapses and has recently been implicated in addiction-related behavior. The ATPase Rpt6 subunit of the 26S proteasome is phosphorylated by Ca 2+ /calmodulin-dependent protein kinases II alpha at ser120 which is thought to regulate proteasome activity and distribution in neurons. Here, we demonstrate that Rpt6 phosphorylation is involved in cocaine-induced locomotor sensitization. Cocaine concomitantly increases proteasome activity and Rpt6 S120 phosphorylation in cultured neurons and in various brain regions of wild type mice including the nucleus accumbens and prefrontal cortex. In contrast, cocaine does not increase proteasome activity in Rpt6 phospho-mimetic (ser120Asp) mice. Strikingly, we found a complete absence of cocaine-induced locomotor sensitization in the Rpt6 ser120Asp mice. Together, these findings suggest a critical role for Rpt6 phosphorylation and proteasome function in the regulation cocaine-induced behavioral plasticity. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Pepper, Sweet (Capsicum annuum)

    NARCIS (Netherlands)

    Heidmann, I.; Boutilier, K.A.

    2015-01-01

    Capsicum (pepper) species are economically important crops that are recalcitrant to genetic transformation by Agrobacterium ( Agrobacterium tumefaciens ). A number of protocols for pepper transformation have been described but are not routinely applicable. The main bottleneck in pepper

  7. Tau protein degradation is catalyzed by the ATP/ubiquitin-independent 20S proteasome under normal cell conditions

    OpenAIRE

    Grune, Tilman; Botzen, Diana; Engels, Martina; Voss, Peter; Kaiser, Barbara; Jung, Tobias; Grimm, Stefanie; Ermak, Gennady; Davies, Kelvin J. A.

    2010-01-01

    Tau is the major protein exhibiting intracellular accumulation in Alzheimer disease. The mechanisms leading to its accumulation are not fully understood. It has been proposed that the proteasome is responsible for degrading tau but, since proteasomal inhibitors block both the ubiquitin-dependent 26S proteasome and the ubiqutin-independent 20S proteasome pathways, it is not clear which of these pathways is involved in tau degradation. Some involvement of the ubiquitin ligase, CHIP in tau degra...

  8. Proteasomes in the archaea: from structure to function.

    Science.gov (United States)

    Maupin-Furlow, J A; Wilson, H L; Kaczowka, S J; Ou, M S

    2000-09-01

    Survival of cells is critically dependent on their ability to rapidly adapt to changes in the natural environment no matter how 'extreme'the habitat. An interplay between protein folding and hydrolysis is emerging as a central mechanism for stress survival and proper cell function. In eucaryotic cells, most proteins destined for destruction are covalently modified by the ubiquitin-system and then degraded in an energy-dependent mechanism by the 26S proteasome, a multicatalytic protease. The 26S proteasome is composed of a 20S proteolytic core and 19S cap (PA700) regulator which includes six AAA+ ATPase subunits. Related AAA+ proteins and 20S proteasomes are found in the archaea and Gram positive actinomycetes. In general, 20S proteasomes form a barrel-shaped nanocompartment with narrow openings which isolate rather non-specific proteolytic active-sites to the interior of the cylinder and away from interaction with cytosolic proteins. The proteasome-associated AAA+ proteins are predicted to form ring-like structures which unfold substrate proteins for entry into the central proteolytic 20S chamber resulting in an energy-dependent and processive destruction of the protein. Detailed biochemical and biophysical analysis as well as identification of proteasomes in archaea with developed genetic tools are providing a foundation for understanding the biological role of the proteasome in these unusual organisms.

  9. Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation.

    Science.gov (United States)

    Boehringer, Jonas; Riedinger, Christiane; Paraskevopoulos, Konstantinos; Johnson, Eachan O D; Lowe, Edward D; Khoudian, Christina; Smith, Dominique; Noble, Martin E M; Gordon, Colin; Endicott, Jane A

    2012-11-15

    The ubiquitin-proteasome system targets selected proteins for degradation by the 26S proteasome. Rpn12 is an essential component of the 19S regulatory particle and plays a role in recruiting the extrinsic ubiquitin receptor Rpn10. In the present paper we report the crystal structure of Rpn12, a proteasomal PCI-domain-containing protein. The structure helps to define a core structural motif for the PCI domain and identifies potential sites through which Rpn12 might form protein-protein interactions. We demonstrate that mutating residues at one of these sites impairs Rpn12 binding to Rpn10 in vitro and reduces Rpn10 incorporation into proteasomes in vivo.

  10. Interplay between Molecular Chaperones and the Ubiquitin-Proteasome System in Targeting of Misfolded Proteins for Degradation

    DEFF Research Database (Denmark)

    Poulsen, Esben Guldahl

    interacting with purified 26S proteasomes, and the subsequent characterization of two novel proteasome interacting proteins. The third study was aimed at analyzing the chaperone-assisted pathway leading to degradation of misfolded kinetochore proteins in S. pombe. In this study chaperones, E2s, E3s and DUBs...

  11. Chemotherapy inhibits skeletal muscle ubiquitin-proteasome-dependent proteolysis.

    Science.gov (United States)

    Tilignac, Thomas; Temparis, Sandrine; Combaret, Lydie; Taillandier, Daniel; Pouch, Marie-Noëlle; Cervek, Matjaz; Cardenas, Diana M; Le Bricon, Thierry; Debiton, Eric; Samuels, Susan E; Madelmont, Jean-Claude; Attaix, Didier

    2002-05-15

    Chemotherapy has cachectic effects, but it is unknown whether cytostatic agents alter skeletal muscle proteolysis. We hypothesized that chemotherapy-induced alterations in protein synthesis should result in the increased incidence of abnormal proteins, which in turn should stimulate ubiquitin-proteasome-dependent proteolysis. The effects of the nitrosourea cystemustine were investigated in skeletal muscles from both healthy and colon 26 adenocarcinoma-bearing mice, an appropriate model for testing the impact of cytostatic agents. Muscle wasting was seen in both groups of mice 4 days after a single cystemustine injection, and the drug further increased the loss of muscle proteins already apparent in tumor-bearing animals. Cystemustine cured the tumor-bearing mice with 100% efficacy. Surprisingly, within 11 days of treatment, rates of muscle proteolysis progressively decreased below basal levels observed in healthy control mice and contributed to the cessation of muscle wasting. Proteasome-dependent proteolysis was inhibited by mechanisms that include reduced mRNA levels for 20S and 26S proteasome subunits, decreased protein levels of 20S proteasome subunits and the S14 non-ATPase subunit of the 26S proteasome, and impaired chymotrypsin- and trypsin-like activities of the enzyme. A combination of cisplatin and ifosfamide, two drugs that are widely used in the treatment of cancer patients, also depressed the expression of proteasomal subunits in muscles from rats bearing the MatB adenocarcinoma below basal levels. Thus, a down-regulation of ubiquitin-proteasome-dependent proteolysis is observed with various cytostatic agents and contributes to reverse the chemotherapy-induced muscle wasting.

  12. The 20S proteasome as an assembly platform for the 19S regulatory complex

    DEFF Research Database (Denmark)

    Hendil, Klaus Aksel Bjørner; Kriegenburg, Franziska; Tanaka, Keiji

    2009-01-01

    26S proteasomes consist of cylindrical 20S proteasomes with 19S regulatory complexes attached to the ends. Treatment with high concentrations of salt causes the regulatory complexes to separate into two sub-complexes, the base, which is in contact with the 20S proteasome, and the lid, which...... is the distal part of the 19S complex. Here, we describe two assembly intermediates of the human regulatory complex. One is a dimer of the two ATPase subunits, Rpt3 and Rpt6. The other is a complex of nascent Rpn2, Rpn10, Rpn11, Rpn13, and Txnl1, attached to preexisting 20S proteasomes. This early assembly...... complex does not yet contain Rpn1 or any of the ATPase subunits of the base. Thus, assembly of 19S regulatory complexes takes place on preexisting 20S proteasomes, and part of the lid is assembled before the base....

  13. ABA-dependent inhibition of the ubiquitin proteasome system during germination at high temperature in Arabidopsis.

    Science.gov (United States)

    Chiu, Rex Shun; Pan, Shiyue; Zhao, Rongmin; Gazzarrini, Sonia

    2016-12-01

    During germination, endogenous and environmental factors trigger changes in the transcriptome, translatome and proteome to break dormancy. In Arabidopsis thaliana, the ubiquitin proteasome system (UPS) degrades proteins that promote dormancy to allow germination. While research on the UPS has focused on the identification of proteasomal substrates, little information is known about the regulation of its activity. Here we characterized the activity of the UPS during dormancy release and maintenance by monitoring protein ubiquitination and degradation of two proteasomal substrates: Suc-LLVY-AMC, a well characterized synthetic substrate, and FUSCA3 (FUS3), a dormancy-promoting transcription factor degraded by the 26S proteasome. Our data indicate that proteasome activity and protein ubiquitination increase during imbibition at optimal temperature (21°C), and are required for seed germination. However, abscisic acid (ABA) and supraoptimal temperature (32°C) inhibit germination by dampening both protein ubiquitination and proteasome activity. Inhibition of UPS function by high temperature is reduced by the ABA biosynthesis inhibitor, fluridone, and in ABA biosynthetic mutants, suggesting that it is ABA dependent. Accordingly, inhibition of FUS3 degradation at 32°C is also dependent on ABA. Native gels show that inhibition of proteasome activity is caused by interference with the 26S/30S ratio as well as free 19S and 20S levels, impacting the proteasome degradation cycle. Transfer experiments show that ABA-mediated inhibition of proteasome activity at 21°C is restricted to the first 2 days of germination, a time window corresponding to seed sensitivity to environmental and ABA-mediated growth inhibition. Our data show that ABA and high temperature inhibit germination under unfavourable growth conditions by repressing the UPS. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

  14. Peptide-based proteasome inhibitors in anticancer drug design.

    Science.gov (United States)

    Micale, Nicola; Scarbaci, Kety; Troiano, Valeria; Ettari, Roberta; Grasso, Silvana; Zappalà, Maria

    2014-09-01

    The identification of the key role of the eukaryotic 26S proteasome in regulated intracellular proteolysis and its importance as a target in many pathological conditions wherein the proteasomal activity is defective (e.g., malignancies, autoimmune diseases, neurodegenerative diseases, etc.) prompted several research groups to the development of specific inhibitors of this multicatalytic complex with the aim of obtaining valid drug candidates. In regard to the anticancer therapy, the peptide boronate bortezomib (Velcade®) represents the first molecule approved by FDA for the treatment of multiple myeloma in 2003 and mantle cell lymphoma in 2006. Since then, a plethora of molecules targeting the proteasome have been identified as potential anticancer agents and a few of them reached clinical trials or are already in the market (i.e., carfilzomib; Kyprolis®). In most cases, the design of new proteasome inhibitors (PIs) takes into account a proven peptide or pseudopeptide motif as a base structure and places other chemical entities throughout the peptide skeleton in such a way to create an efficacious network of interactions within the catalytic sites. The purpose of this review is to provide an in-depth look at the current state of the research in the field of peptide-based PIs, specifically those ones that might find an application as anticancer agents. © 2014 Wiley Periodicals, Inc.

  15. E1AF degradation by a ubiquitin-proteasome pathway

    International Nuclear Information System (INIS)

    Takahashi, Akiko; Higashino, Fumihiro; Aoyagi, Mariko; Yoshida, Koichi; Itoh, Miyuki; Kobayashi, Masanobu; Totsuka, Yasunori; Kohgo, Takao; Shindoh, Masanobu

    2005-01-01

    E1AF is a member of the ETS family of transcription factors. In mammary tumors, overexpression of E1AF is associated with tumorigenesis, but E1AF protein has hardly been detected and its degradation mechanism is not yet clear. Here we show that E1AF protein is stabilized by treatment with the 26S protease inhibitor MG132. We found that E1AF was modified by ubiquitin through the C-terminal region and ubiquitinated E1AF aggregated in nuclear dots, and that the inhibition of proteasome-activated transcription from E1AF target promoters. These results suggest that E1AF is degraded via the ubiquitin-proteasome pathway, which has some effect on E1AF function

  16. Bladder cancer detection using a peptide substrate of the 20S proteasome.

    Science.gov (United States)

    Gruba, Natalia; Wysocka, Magdalena; Brzezińska, Magdalena; Dębowski, Dawid; Sieńczyk, Marcin; Gorodkiewicz, Ewa; Guszcz, Tomasz; Czaplewski, Cezary; Rolka, Krzysztof; Lesner, Adam

    2016-08-01

    The 20S catalytic core of the human 26S proteasome can be secreted from cells, and high levels of extracellular 20S proteasome have been linked to many types of cancers and autoimmune diseases. Several diagnostic approaches have been developed that detect 20S proteasome activity in plasma, but these suffer from problems with efficiency and sensitivity. In this report, we describe the optimization and synthesis of an internally quenched fluorescent substrate of the 20S proteasome, and investigate its use as a potential diagnostic test in bladder cancer. This peptide, 2-aminobenzoic acid (ABZ)-Val-Val-Ser-Tyr-Ala-Met-Gly-Tyr(3-NO2 )-NH2 , is cleaved by the chymotrypsin 20S proteasome subunit and displays an excellent specificity constant value (9.7 × 10(5)  m(-1) ·s(-1) ) and a high kcat (8 s(-1) ). Using this peptide, we identified chymotrypsin-like proteasome activity in the majority of urine samples obtained from patients with bladder cancer, whereas the proteasome activity in urine samples from healthy volunteers was below the detection limit (0.5 pm). These findings were confirmed by an inhibitory study and immunochemistry methods. © 2016 Federation of European Biochemical Societies.

  17. Aleurotrachelus trachoides (pepper whitefly)

    Science.gov (United States)

    Aleurotrachelus trachoides Back also known as solanum or pepper whitefly is a new addition to the list of serious whitefly pests found in Florida. According to EPPO global database, it is a pest of over 70 different crops worldwide, which include a combination of edibles, ornamentals, palms, and wee...

  18. Apoptosis and radiosensitization of Hodgkin cells by proteasome inhibition

    International Nuclear Information System (INIS)

    Pajonk, Frank; Pajonk, Katja; McBride, William H.

    2000-01-01

    Purpose: Malignant cells from Hodgkin's disease have been reported to be defective in regulation of NF-κB activity. Ionizing radiation is known to activate NF-κB, and it has been suggested that this pathway may protect cells from apoptosis following exposure to radiation and other therapeutic agents. Defective NF-κB regulation in Hodgkin cells could therefore dictate the response of this disease to therapy, as well as be responsible for maintaining the malignant phenotype. The purpose of this study was to explore whether NF-κB activity could be modulated in Hodgkin cells and whether it determines the response of these cells to treatment with ionizing radiation and/or dexamethasone. Methods and Materials: Activation of NF-κB in cells is accomplished in large part by degradation of its inhibitor IκB through the 26s proteasome. HD-My-Z Hodgkin cells were treated with the proteasome inhibitor MG-132 or transduced with a dominant negative super-repressor IκBα. Clonogenic survival, apoptosis, proteasome activity, and NF-κB binding activity were monitored in response to ionizing radiation and/or dexamethasone treatment. Results: HD-My-Z Hodgkin cells had modest NF-κB levels but, unlike other cell types, did not decrease their level of constitutively active NF-κB in response to proteasome inhibition with MG-132. In contrast, transduction with a non-phosphorable IκBα construct abolished expression. MG-132 did, however, induce apoptosis in HD-My-Z cells and sensitized them to ionizing radiation. Dexamethasone treatment had no effect on NF-κB activity or clonogenic survival of Hodgkin cells, but protected them from irradiation. Conclusion: We conclude that inhibition of 26s proteasome activity can induce apoptosis in HD-My-Z Hodgkin cells and radiosensitize them, in spite of the fact that their constitutively active NF-κB levels are unaltered. The proteasome may be a promising new therapeutic target for intervention in this disease. In contrast, the use of

  19. Conformational switching in the coiled-coil domains of a proteasomal ATPase regulates substrate processing.

    Science.gov (United States)

    Snoberger, Aaron; Brettrager, Evan J; Smith, David M

    2018-06-18

    Protein degradation in all domains of life requires ATPases that unfold and inject proteins into compartmentalized proteolytic chambers. Proteasomal ATPases in eukaryotes and archaea contain poorly understood N-terminally conserved coiled-coil domains. In this study, we engineer disulfide crosslinks in the coiled-coils of the archaeal proteasomal ATPase (PAN) and report that its three identical coiled-coil domains can adopt three different conformations: (1) in-register and zipped, (2) in-register and partially unzipped, and (3) out-of-register. This conformational heterogeneity conflicts with PAN's symmetrical OB-coiled-coil crystal structure but resembles the conformational heterogeneity of the 26S proteasomal ATPases' coiled-coils. Furthermore, we find that one coiled-coil can be conformationally constrained even while unfolding substrates, and conformational changes in two of the coiled-coils regulate PAN switching between resting and active states. This switching functionally mimics similar states proposed for the 26S proteasome from cryo-EM. These findings thus build a mechanistic framework to understand regulation of proteasome activity.

  20. Direct ubiquitin independent recognition and degradation of a folded protein by the eukaryotic proteasomes-origin of intrinsic degradation signals.

    Directory of Open Access Journals (Sweden)

    Amit Kumar Singh Gautam

    Full Text Available Eukaryotic 26S proteasomes are structurally organized to recognize, unfold and degrade globular proteins. However, all existing model substrates of the 26S proteasome in addition to ubiquitin or adaptor proteins require unstructured regions in the form of fusion tags for efficient degradation. We report for the first time that purified 26S proteasome can directly recognize and degrade apomyoglobin, a globular protein, in the absence of ubiquitin, extrinsic degradation tags or adaptor proteins. Despite a high affinity interaction, absence of a ligand and presence of only helices/loops that follow the degradation signal, apomyoglobin is degraded slowly by the proteasome. A short floppy F-helix exposed upon ligand removal and in conformational equilibrium with a disordered structure is mandatory for recognition and initiation of degradation. Holomyoglobin, in which the helix is buried, is neither recognized nor degraded. Exposure of the floppy F-helix seems to sensitize the proteasome and primes the substrate for degradation. Using peptide panning and competition experiments we speculate that initial encounters through the floppy helix and additional strong interactions with N-terminal helices anchors apomyoglobin to the proteasome. Stabilizing helical structure in the floppy F-helix slows down degradation. Destabilization of adjacent helices accelerates degradation. Unfolding seems to follow the mechanism of helix unraveling rather than global unfolding. Our findings while confirming the requirement for unstructured regions in degradation offers the following new insights: a origin and identification of an intrinsic degradation signal in the substrate, b identification of sequences in the native substrate that are likely to be responsible for direct interactions with the proteasome, and c identification of critical rate limiting steps like exposure of the intrinsic degron and destabilization of an unfolding intermediate that are presumably

  1. Regulators of the proteasome pathway, Uch37 and Rpn13, play distinct roles in mouse development.

    Directory of Open Access Journals (Sweden)

    Amin Al-Shami

    Full Text Available Rpn13 is a novel mammalian proteasomal receptor that has recently been identified as an amplification target in ovarian cancer. It can interact with ubiquitin and activate the deubiquitinating enzyme Uch37 at the 26S proteasome. Since neither Rpn13 nor Uch37 is an integral proteasomal subunit, we explored whether either protein is essential for mammalian development and survival. Deletion of Uch37 resulted in prenatal lethality in mice associated with severe defect in embryonic brain development. In contrast, the majority of Rpn13-deficient mice survived to adulthood, although they were smaller at birth and fewer in number than wild-type littermates. Absence of Rpn13 produced tissue-specific effects on proteasomal function: increased proteasome activity in adrenal gland and lymphoid organs, and decreased activity in testes and brain. Adult Rpn13(-/- mice reached normal body weight but had increased body fat content and were infertile due to defective gametogenesis. Additionally, Rpn13(-/- mice showed increased T-cell numbers, resembling growth hormone-mediated effects. Indeed, serum growth hormone and follicular stimulating hormone levels were significantly increased in Rpn13(-/- mice, while growth hormone receptor expression was reduced in the testes. In conclusion, this is the first report characterizing the physiological roles of Uch37 and Rpn13 in murine development and implicating a non-ATPase proteasomal protein, Rpn13, in the process of gametogenesis.

  2. Occurrence of pepper mild mottle virus in greenhousegrown pepper ...

    African Journals Online (AJOL)

    Severe systemic viral symptoms were observed on the leaves of infected pepper (Capsicum annuum L.) plants cultivated in Antalya located in the Mediterranean coast of Turkey, in 2008. The symptoms on the diseased pepper plants included, mosaic, mottle, chlorosis coupled with stunting, chlorotic spots, distortion of the ...

  3. Divergent tissue and sex effects of rapamycin on the proteasome-chaperone network of old mice

    Directory of Open Access Journals (Sweden)

    Karl Andrew Rodriguez

    2014-11-01

    Full Text Available Rapamycin, an allosteric inhibitor of the mTOR kinase, increases longevity in mice in a sex-specific manner. In contrast to the widely accepted theory that a loss of proteasome activity is detrimental to both life- and healthspan, biochemical studies in vitro reveal that rapamycin inhibits 20S proteasome peptidase activity. We tested if this unexpected finding is also evident after chronic rapamycin treatment in vivo by measuring peptidase activities for both the 26S and 20S proteasome in liver, fat, and brain tissues of old, male and female mice fed encapsulated chow containing 2.24mg/kg (14 ppm rapamycin for 6 months. Further we assessed if rapamycin altered expression of the chaperone proteins known to interact with the proteasome-mediated degradation system (PMDS, heat shock factor 1 (HSF1, and the levels of key mTOR pathway proteins. Rapamycin had little effect on liver proteasome activity in either gender, but increased proteasome activity in female brain lysates and lowered its activity in female fat tissue. Rapamycin-induced changes in molecular chaperone levels were also more substantial in tissues from female animals. Furthermore, mTOR pathway proteins showed more significant changes in female tissues compared to those from males. These data show collectively that there are divergent tissue and sex effects of rapamycin on the proteasome-chaperone network and that these may be linked to the disparate effects of rapamycin on males and females. Further our findings suggest that rapamycin induces indirect regulation of the PMDS/heat-shock response through its modulation of the mTOR pathway rather than via direct interactions between rapamycin and the proteasome.

  4. Murraya koenigii leaf extract inhibits proteasome activity and induces cell death in breast cancer cells

    Directory of Open Access Journals (Sweden)

    Noolu Bindu

    2013-01-01

    Full Text Available Abstract Background Inhibition of the proteolytic activity of 26S proteasome, the protein-degrading machine, is now considered a novel and promising approach for cancer therapy. Interestingly, proteasome inhibitors have been demonstrated to selectively kill cancer cells and also enhance the sensitivity of tumor cells to chemotherapeutic agents. Recently, polyphenols/flavonoids have been reported to inhibit proteasome activity. Murraya koenigii Spreng, a medicinally important herb of Indian origin, has been used for centuries in the Ayurvedic system of medicine. Here we show that Murraya koenigii leaves (curry leaves, a rich source of polyphenols, inhibit the proteolytic activity of the cancer cell proteasome, and cause cell death. Methods Hydro-methanolic extract of curry leaves (CLE was prepared and its total phenolic content [TPC] determined by, the Folin-Ciocalteau’s method. Two human breast carcinoma cell lines: MCF-7 and MDA-MB-231 and a normal human lung fibroblast cell line, WI-38 were used for the studies. Cytotoxicity of the CLE was assessed by the MTT assay. We studied the effect of CLE on growth kinetics using colony formation assay. Growth arrest was assessed by cell cycle analysis and apoptosis by Annexin-V binding using flow cytometry. Inhibition of the endogenous 26S proteasome was studied in intact cells and cell extracts using substrates specific to 20S proteasomal enzymes. Results CLE decreased cell viability and altered the growth kinetics in both the breast cancer cell lines in a dose-dependent manner. It showed a significant arrest of cells in the S phase albeit in cancer cells only. Annexin V binding data suggests that cell death was via the apoptotic pathway in both the cancer cell lines. CLE treatment significantly decreased the activity of the 26S proteasome in the cancer but not normal cells. Conclusions Our study suggests M. koenigii leaves to be a potent source of proteasome inhibitors that lead to cancer cell death

  5. Murraya koenigii leaf extract inhibits proteasome activity and induces cell death in breast cancer cells.

    Science.gov (United States)

    Noolu, Bindu; Ajumeera, Rajanna; Chauhan, Anitha; Nagalla, Balakrishna; Manchala, Raghunath; Ismail, Ayesha

    2013-01-09

    Inhibition of the proteolytic activity of 26S proteasome, the protein-degrading machine, is now considered a novel and promising approach for cancer therapy. Interestingly, proteasome inhibitors have been demonstrated to selectively kill cancer cells and also enhance the sensitivity of tumor cells to chemotherapeutic agents. Recently, polyphenols/flavonoids have been reported to inhibit proteasome activity. Murraya koenigii Spreng, a medicinally important herb of Indian origin, has been used for centuries in the Ayurvedic system of medicine. Here we show that Murraya koenigii leaves (curry leaves), a rich source of polyphenols, inhibit the proteolytic activity of the cancer cell proteasome, and cause cell death. Hydro-methanolic extract of curry leaves (CLE) was prepared and its total phenolic content [TPC] determined by, the Folin-Ciocalteau's method. Two human breast carcinoma cell lines: MCF-7 and MDA-MB-231 and a normal human lung fibroblast cell line, WI-38 were used for the studies. Cytotoxicity of the CLE was assessed by the MTT assay. We studied the effect of CLE on growth kinetics using colony formation assay. Growth arrest was assessed by cell cycle analysis and apoptosis by Annexin-V binding using flow cytometry. Inhibition of the endogenous 26S proteasome was studied in intact cells and cell extracts using substrates specific to 20S proteasomal enzymes. CLE decreased cell viability and altered the growth kinetics in both the breast cancer cell lines in a dose-dependent manner. It showed a significant arrest of cells in the S phase albeit in cancer cells only. Annexin V binding data suggests that cell death was via the apoptotic pathway in both the cancer cell lines. CLE treatment significantly decreased the activity of the 26S proteasome in the cancer but not normal cells. Our study suggests M. koenigii leaves to be a potent source of proteasome inhibitors that lead to cancer cell death. Therefore, identification of active component(s) from the leaf

  6. Fission yeast 26S proteasome mutants are multi-drug resistant due to stabilization of the Pap1transcription factor

    DEFF Research Database (Denmark)

    Penney, Mary; Samejima, Itaru; Wilkinson, Caroline

    2012-01-01

    Here we report the result of a genetic screen for mutants resistant to the microtubule poison methyl benzimidazol-2-yl carbamate (MBC) that were also temperature sensitive for growth. In total the isolated mutants were distributed in ten complementation groups. Cloning experiments revealed...

  7. The ubiquitin-proteasome system

    Indian Academy of Sciences (India)

    ... the discovery of protein ubiquitination has led to the recognition of cellular proteolysis as a central area of research in biology. Eukaryotic proteins targeted for degradation by this pathway are first 'tagged' by multimers of a protein known as ubiquitin and are later proteolyzed by a giant enzyme known as the proteasome.

  8. Decontamination of Black Pepper (Pepper Negrum) in grains by irradiation

    International Nuclear Information System (INIS)

    Alvarez, M.; Pino, J.; Zayas, M.

    1991-01-01

    The effect of irradiation with doses ranging between 3 and 10 kGy on the microbiological quality, on different chemical components and on the organoleptic quality of black pepper in grains was studied in this paper. The results showed the effectiveness of treatment in the decontamination of this spice. Piperin content, yield of essential oil and its composition as well as the organoleptic quality did not change with applied doses to black pepper in grains

  9. Altered composition of liver proteasome assemblies contributes to enhanced proteasome activity in the exceptionally long-lived naked mole-rat.

    Science.gov (United States)

    Rodriguez, Karl A; Edrey, Yael H; Osmulski, Pawel; Gaczynska, Maria; Buffenstein, Rochelle

    2012-01-01

    The longest-lived rodent, the naked mole-rat (Bathyergidae; Heterocephalus glaber), maintains robust health for at least 75% of its 32 year lifespan, suggesting that the decline in genomic integrity or protein homeostasis routinely observed during aging, is either attenuated or delayed in this extraordinarily long-lived species. The ubiquitin proteasome system (UPS) plays an integral role in protein homeostasis by degrading oxidatively-damaged and misfolded proteins. In this study, we examined proteasome activity in naked mole-rats and mice in whole liver lysates as well as three subcellular fractions to probe the mechanisms behind the apparently enhanced effectiveness of UPS. We found that when compared with mouse samples, naked mole-rats had significantly higher chymotrypsin-like (ChT-L) activity and a two-fold increase in trypsin-like (T-L) in both whole lysates as well as cytosolic fractions. Native gel electrophoresis of the whole tissue lysates showed that the 20S proteasome was more active in the longer-lived species and that 26S proteasome was both more active and more populous. Western blot analyses revealed that both 19S subunits and immunoproteasome catalytic subunits are present in greater amounts in the naked mole-rat suggesting that the observed higher specific activity may be due to the greater proportion of immunoproteasomes in livers of healthy young adults. It thus appears that proteasomes in this species are primed for the efficient removal of stress-damaged proteins. Further characterization of the naked mole-rat proteasome and its regulation could lead to important insights on how the cells in these animals handle increased stress and protein damage to maintain a longer health in their tissues and ultimately a longer life.

  10. Altered composition of liver proteasome assemblies contributes to enhanced proteasome activity in the exceptionally long-lived naked mole-rat.

    Directory of Open Access Journals (Sweden)

    Karl A Rodriguez

    Full Text Available The longest-lived rodent, the naked mole-rat (Bathyergidae; Heterocephalus glaber, maintains robust health for at least 75% of its 32 year lifespan, suggesting that the decline in genomic integrity or protein homeostasis routinely observed during aging, is either attenuated or delayed in this extraordinarily long-lived species. The ubiquitin proteasome system (UPS plays an integral role in protein homeostasis by degrading oxidatively-damaged and misfolded proteins. In this study, we examined proteasome activity in naked mole-rats and mice in whole liver lysates as well as three subcellular fractions to probe the mechanisms behind the apparently enhanced effectiveness of UPS. We found that when compared with mouse samples, naked mole-rats had significantly higher chymotrypsin-like (ChT-L activity and a two-fold increase in trypsin-like (T-L in both whole lysates as well as cytosolic fractions. Native gel electrophoresis of the whole tissue lysates showed that the 20S proteasome was more active in the longer-lived species and that 26S proteasome was both more active and more populous. Western blot analyses revealed that both 19S subunits and immunoproteasome catalytic subunits are present in greater amounts in the naked mole-rat suggesting that the observed higher specific activity may be due to the greater proportion of immunoproteasomes in livers of healthy young adults. It thus appears that proteasomes in this species are primed for the efficient removal of stress-damaged proteins. Further characterization of the naked mole-rat proteasome and its regulation could lead to important insights on how the cells in these animals handle increased stress and protein damage to maintain a longer health in their tissues and ultimately a longer life.

  11. EFFECT OF ALLIGATOR PEPPER (ZINGIBERACEAE ...

    African Journals Online (AJOL)

    Daniel Owu

    proven fertility from a pilot study were randomly paired in fifteen cages in a well ventillated room. After three days of ... Key words : Effect, Alligator pepper, Herbal remedy, Nutrition, Pregnancy. Introduction ... ad libitum. Experimental drug.

  12. Resistance to thrips in pepper

    NARCIS (Netherlands)

    Maharijaya, A.

    2013-01-01

    Pepper (Capsicum) production is constrained by heavy infestations of thrips, causing direct and indirect (by transmitting viruses) damage. Thrips control using chemical insecticides, biological agents, culture practices and integrated pest management has limited success. The

  13. Mutation breeding in pepper

    Energy Technology Data Exchange (ETDEWEB)

    Daskalov, S [Plant Breeding Unit, Joint FAO/IAEA Division of Isotope and Radiation Applications of Atomic Energy for Food and Agricultural Development, Seibersdorf Laboratory, International Atomic Energy Agency, Vienna (Austria)

    1986-03-01

    Pepper (Capsicum sp.) is an important vegetable and spice crop widely grown in tropical as well as in temperate regions. Until recently the improvement programmes were based mainly on using natural sources of germ plasma, crossbreeding and exploiting the heterosis of F{sub 1} hybrids. However, interest in using induced mutations is growing. A great number of agronomically useful mutants as well as mutants valuable for genetic, cytological and physiological studies have been induced and described. In this review information is presented about suitable mutagen treatment procedures with radiation as well as chemicals, M{sub 1} effects, handling the treated material in M{sub 1}, M{sub 2} and subsequent generations, and mutant screening procedures. This is supplemented by a description of reported useful mutants and released cultivars. Finally, general advice is given on when and how to incorporate mutation induction in Capsicum improvement programmes. (author)

  14. Mutation breeding in pepper

    International Nuclear Information System (INIS)

    Daskalov, S.

    1986-01-01

    Pepper (Capsicum sp.) is an important vegetable and spice crop widely grown in tropical as well as in temperate regions. Until recently the improvement programmes were based mainly on using natural sources of germ plasma, crossbreeding and exploiting the heterosis of F 1 hybrids. However, interest in using induced mutations is growing. A great number of agronomically useful mutants as well as mutants valuable for genetic, cytological and physiological studies have been induced and described. In this review information is presented about suitable mutagen treatment procedures with radiation as well as chemicals, M 1 effects, handling the treated material in M 1 , M 2 and subsequent generations, and mutant screening procedures. This is supplemented by a description of reported useful mutants and released cultivars. Finally, general advice is given on when and how to incorporate mutation induction in Capsicum improvement programmes. (author)

  15. The proteasome inhibitor MG-132 sensitizes PC-3 prostate cancer cells to ionizing radiation by a DNA-PK-independent mechanism

    International Nuclear Information System (INIS)

    Pajonk, Frank; Ophoven, Arndt van; Weissenberger, Christian; McBride, William H

    2005-01-01

    By modulating the expression levels of specific signal transduction molecules, the 26S proteasome plays a central role in determining cell cycle progression or arrest and cell survival or death in response to stress stimuli, including ionizing radiation. Inhibition of proteasome function by specific drugs results in cell cycle arrest, apoptosis and radiosensitization of many cancer cell lines. This study investigates whether there is also a concomitant increase in cellular radiosensitivity if proteasome inhibition occurs only transiently before radiation. Further, since proteasome inhibition has been shown to activate caspase-3, which is involved in apoptosis, and caspase-3 can cleave DNA-PKcs, which is involved in DNA-double strand repair, the hypothesis was tested that caspase-3 activation was essential for both apoptosis and radiosensitization following proteasome inhibition. Prostate carcinoma PC-3 cells were treated with the reversible proteasome inhibitor MG-132. Cell cycle distribution, apoptosis, caspase-3 activity, DNA-PKcs protein levels and DNA-PK activity were monitored. Radiosensitivity was assessed using a clonogenic assay. Inhibition of proteasome function caused cell cycle arrest and apoptosis but this did not involve early activation of caspase-3. Short-time inhibition of proteasome function also caused radiosensitization but this did not involve a decrease in DNA-PKcs protein levels or DNA-PK activity. We conclude that caspase-dependent cleavage of DNA-PKcs during apoptosis does not contribute to the radiosensitizing effects of MG-132

  16. Proteasomes remain intact, but show early focal alteration in their composition in a mouse model of amyotrophic lateral sclerosis.

    Science.gov (United States)

    Kabashi, Edor; Agar, Jeffrey N; Hong, Yu; Taylor, David M; Minotti, Sandra; Figlewicz, Denise A; Durham, Heather D

    2008-06-01

    In amyotrophic lateral sclerosis caused by mutations in Cu/Zn-superoxide dismutase (SOD1), altered solubility and aggregation of the mutant protein implicates failure of pathways for detecting and catabolizing misfolded proteins. Our previous studies demonstrated early reduction of proteasome-mediated proteolytic activity in lumbar spinal cord of SOD1(G93A) transgenic mice, tissue particularly vulnerable to disease. The purpose of this study was to identify any underlying abnormalities in proteasomal structure. In lumbar spinal cord of pre-symptomatic mice [postnatal day 45 (P45) and P75], normal levels of structural 20S alpha subunits were incorporated into 20S/26S proteasomes; however, proteasomal complexes separated by native gel electrophoresis showed decreased immunoreactivity with antibodies to beta3, a structural subunit of the 20S proteasome core, and beta5, the subunit with chymotrypsin-like activity. This occurred prior to increase in beta5i immunoproteasomal subunit. mRNA levels were maintained and no association of mutant SOD1 with proteasomes was identified, implicating post-transcriptional mechanisms. mRNAs also were maintained in laser captured motor neurons at a later stage of disease (P100) in which multiple 20S proteins are reduced relative to the surrounding neuropil. Increase in detergent-insoluble, ubiquitinated proteins at P75 provided further evidence of stress on mechanisms of protein quality control in multiple cell types prior to significant motor neuron death.

  17. Features of proteasome functioning in malignant tumors

    Science.gov (United States)

    Kondakova, I. V.; Spirina, L. V.; Shashova, E. E.; Kolegova, E. S.; Slonimskaya, E. M.; Kolomiets, L. A.; Afanas'ev, S. G.; Choinzonov, Y. L.

    2017-09-01

    Proteasome ubiquitin system is the important system of intracellular proteolysis. The activity of the proteasomes may undergo changes during cancer development. We studied the chymotrypsin-like activity of proteasomes, their subunit composition, and their association with tumor stage in breast cancer, head and neck squamous cell carcinoma, endometrial cancer, renal cancer, bladder cancer, stomach cancer, ovarian cancer, and colorectal cancer. The increase in chymotrypsin-like activity of proteasomes and decrease in total proteasome pool compared with adjacent tissues were shown in all malignant tumors excluding kidney cancer. The increase in chymotrypsin-like activity of proteasomes was found in primary tumors with all types of metastasis: lymphogenous of head and neck squamous cell carcinoma, intraperitoneal metastasis of ovarian cancer, hematogenous metastasis colorectal cancer. The exception was kidney cancer, in which there was a decrease in chymotrypsin-like activity with distant metastasis.

  18. Modeling proteasome dynamics in Parkinson's disease

    International Nuclear Information System (INIS)

    Sneppen, Kim; Lizana, Ludvig; Jensen, Mogens H; Pigolotti, Simone; Otzen, Daniel

    2009-01-01

    In Parkinson's disease (PD), there is evidence that α-synuclein (αSN) aggregation is coupled to dysfunctional or overburdened protein quality control systems, in particular the ubiquitin–proteasome system. Here, we develop a simple dynamical model for the on-going conflict between αSN aggregation and the maintenance of a functional proteasome in the healthy cell, based on the premise that proteasomal activity can be titrated out by mature αSN fibrils and their protofilament precursors. In the presence of excess proteasomes the cell easily maintains homeostasis. However, when the ratio between the available proteasome and the αSN protofilaments is reduced below a threshold level, we predict a collapse of homeostasis and onset of oscillations in the proteasome concentration. Depleted proteasome opens for accumulation of oligomers. Our analysis suggests that the onset of PD is associated with a proteasome population that becomes occupied in periodic degradation of aggregates. This behavior is found to be the general state of a proteasome/chaperone system under pressure, and suggests new interpretations of other diseases where protein aggregation could stress elements of the protein quality control system

  19. Analysis of gamma irradiated pepper constituents, 2

    International Nuclear Information System (INIS)

    Takagi, Kazuko; Ochiai, Junko; Okuyama, Tsuneo

    1988-01-01

    Black pepper, white pepper, frozen green pepper and real pink pepper (kept in vinegar) were analyzed by reversed phase IIPLC. The extraction method and HPLC conditions were same as the first report, that is, the extraction from pepper was performed by Automatic Air Hammer and the extracted samples were separated on a reversed phase C 8 column with a concave gradient from 0.1% trifluoro aceticacid (TFA) in water to 75% acetonitrile - 0.1% TFA in water for 60 minutes and detected at 210 nm, 280 nm. The different constituents were observed clearly on chromatogram between black pepper and white pepper. The different constituents were observed between different producing white peppers, and as the result that the analyzed pepper was distinguished its producing district by HPLC chromatogram. In order to investigate of effect of lyophilization on white pepper extracts, lyophilized extraction was analyzed by this HPLC method. Some peaks were decreased by lyophilization. The effect of heat on white pepper constituents was examined. White pepper was heated by electronic oven and thermostat. When the former method was used, decreased peak number (peak height was lower than without heat treatment) was more than latter method. These subtle change was able to be recognized by these HPLC chromatograms. (author)

  20. Decontamination of black pepper and red pepper by gamma radiation

    International Nuclear Information System (INIS)

    Oularbi, S.; Mansouri, B.

    1993-04-01

    In the present work, it has been studied the decontamination of two types of spices (black pepper and red pepper) by gamma radiation. The initial microbial population of spices not treated is about 10 (7) to 10 (8) per gram. The population decrease exponentially with irradiation dose. By this, it has been established that a dose of 6 kGy reduces the microbial flora low than 10 (3) per gram. A total elimination of moulds is obtained at dose of 8 kGy

  1. Cayenne pepper in a meal

    DEFF Research Database (Denmark)

    Andersen, B. V.; Byrne, D. V.; Bredie, W. L.P.

    2017-01-01

    The present study investigated appetite sensations, hedonics, sensory specific desires, physical- and psychological well-being sensations during and after intake of a meal with- and without increased oral heat induced by addition of cayenne pepper. Subjects (n = 66) completed a randomized cross......-over study involving a tomato soup with and without added cayenne pepper (0.593 mg capsaicin). Self-reports were evaluated at 5 min intervals during intake and at 1 h intervals until four hours post intake using VAS-scales and 9-point scales. Sensory specific desires were further studied by liking...... and wanting of food samples representing the sensory profiles: sweet, sour, salt, bitter, fat and spicy, respectively. The soup with cayenne pepper added was perceived significantly more spicy but equally liked, and resulted in significant higher satiation at the end of the meal and one hour post intake...

  2. Inhibition of the host proteasome facilitates papaya ringspot virus accumulation and proteosomal catalytic activity is modulated by viral factor HcPro.

    Directory of Open Access Journals (Sweden)

    Nandita Sahana

    Full Text Available The ubiquitin/26S proteasome system plays an essential role not only in maintaining protein turnover, but also in regulating many other plant responses, including plant-pathogen interactions. Previous studies highlighted different roles of the 20S proteasome in plant defense during virus infection, either indirectly through viral suppressor-mediated degradation of Argonaute proteins, affecting the RNA interference pathway, or directly through modulation of the proteolytic and RNase activity of the 20S proteasome, a component of the 20S proteasome, by viral proteins, affecting the levels of viral proteins and RNAs. Here we show that MG132, a cell permeable proteasomal inhibitor, caused an increase in papaya ringspot virus (PRSV accumulation in its natural host papaya (Carica papaya. We also show that the PRSV HcPro interacts with the papaya homologue of the Arabidopsis PAA (α1 subunit of the 20S proteasome, but not with the papaya homologue of Arabidopsis PAE (α5 subunit of the 20S proteasome, associated with the RNase activity, although the two 20S proteasome subunits interacted with each other. Mutated forms of PRSV HcPro showed that the conserved KITC54 motif in the N-terminal domain of HcPro was necessary for its binding to PAA. Co-agroinfiltration assays demonstrated that HcPro expression mimicked the action of MG132, and facilitated the accumulation of bothtotal ubiquitinated proteins and viral/non-viral exogenous RNA in Nicotiana benthamiana leaves. These effects were not observed by using an HcPro mutant (KITS54, which impaired the HcPro - PAA interaction. Thus, the PRSV HcPro interacts with a proteasomal subunit, inhibiting the action of the 20S proteasome, suggesting that HcPro might be crucial for modulating its catalytic activities in support of virus accumulation.

  3. p53 mutations promote proteasomal activity.

    Science.gov (United States)

    Oren, Moshe; Kotler, Eran

    2016-07-27

    p53 mutations occur very frequently in human cancer. Besides abrogating the tumour suppressive functions of wild-type p53, many of those mutations also acquire oncogenic gain-of-function activities. Augmentation of proteasome activity is now reported as a common gain-of-function mechanism shared by different p53 mutants, which promotes cancer resistance to proteasome inhibitors.

  4. The Xanthomonas campestris type III effector XopJ targets the host cell proteasome to suppress salicylic-acid mediated plant defence.

    Directory of Open Access Journals (Sweden)

    Suayib Üstün

    Full Text Available The phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv requires type III effector proteins (T3Es for virulence. After translocation into the host cell, T3Es are thought to interact with components of host immunity to suppress defence responses. XopJ is a T3E protein from Xcv that interferes with plant immune responses; however, its host cellular target is unknown. Here we show that XopJ interacts with the proteasomal subunit RPT6 in yeast and in planta to inhibit proteasome activity. A C235A mutation within the catalytic triad of XopJ as well as a G2A exchange within the N-terminal myristoylation motif abolishes the ability of XopJ to inhibit the proteasome. Xcv ΔxopJ mutants are impaired in growth and display accelerated symptom development including tissue necrosis on susceptible pepper leaves. Application of the proteasome inhibitor MG132 restored the ability of the Xcv ΔxopJ to attenuate the development of leaf necrosis. The XopJ dependent delay of tissue degeneration correlates with reduced levels of salicylic acid (SA and changes in defence- and senescence-associated gene expression. Necrosis upon infection with Xcv ΔxopJ was greatly reduced in pepper plants with reduced expression of NPR1, a central regulator of SA responses, demonstrating the involvement of SA-signalling in the development of XopJ dependent phenotypes. Our results suggest that XopJ-mediated inhibition of the proteasome interferes with SA-dependent defence response to attenuate onset of necrosis and to alter host transcription. A central role of the proteasome in plant defence is discussed.

  5. The Proteasome Inhibitor MG-132 Protects Hypoxic SiHa Cervical Carcinoma Cells after Cyclic Hypoxia/Reoxygenation from Ionizing Radiation

    Directory of Open Access Journals (Sweden)

    Frank Pajonk

    2006-12-01

    Full Text Available INTRODUCTION: Transient hypoxia and subsequent reoxygenation are common phenomena in solid tumors that greatly influence the outcome of radiation therapy. This study was designed to determine how varying cycles of hypoxia/reoxygenation affect the response of cervical carcinoma cells irradiated under oxic and hypoxic conditions and whether this could be modulated by proteasome inhibition. MATERIALS AND METHODS: Plateau-phase SiHa cervical carcinoma cells in culture were exposed to varying numbers of 30-minute cycles of hypoxia/reoxygenation directly before irradiation under oxic or hypoxic conditions. 26S Proteasome activity was blocked by addition of MG-132. Clonogenic survival was measured by a colonyforming assay. RESULTS: Under oxic conditions, repeated cycles of hypoxia/reoxygenation decreased the clonogenic survival of SiHa cells. This effect was even more pronounced after the inhibition of 26S proteasome complex. In contrast, under hypoxic conditions, SiHa cells were radioresistant, as expected, but this was increased by proteasome inhibition. CONCLUSIONS: Proteasome inhibition radiosensitizes oxygenated tumor cells but may also protect tumor cells from ionizing radiation under certain hypoxic conditions.

  6. Use of proteasome inhibitors in anticancer therapy

    Directory of Open Access Journals (Sweden)

    Sara M. Schmitt

    2011-10-01

    Full Text Available The importance of the ubiquitin-proteasome pathway to cellular function has brought it to the forefront in the search for new anticancer therapies. The ubiquitin-proteasome pathway has proven promising in targeting various human cancers. The approval of the proteasome inhibitor bortezomib for clinical treatment of relapsed/refractory multiple myeloma and mantle cell lymphoma has validated the ubiquitin-proteasome as a rational target. Bortezomib has shown positive results in clinical use but some toxicity and side effects, as well as resistance, have been observed, indicating that further development of novel, less toxic drugs is necessary. Because less toxic drugs are necessary and drug development can be expensive and time-consuming, using existing drugs that can target the ubiquitin-proteasome pathway in new applications, such as cancer therapy, may be effective in expediting the regulatory process and bringing new drugs to the clinic. Toward this goal, previously approved drugs, such as disulfiram, as well as natural compounds found in common foods, such as green tea polyphenol (--EGCG and the flavonoid apigenin, have been investigated for their possible proteasome inhibitory and cell death inducing abilities. These compounds proved quite promising in preclinical studies and have now moved into clinical trials, with preliminary results that are encouraging. In addition to targeting the catalytic activity of the proteasome pathway, upstream regulators, such as the 19S regulatory cap, as well as E1, E2, and E3, are now being investigated as potential drug targets. This review outlines the development of novel proteasome inhibitors from preclinical to clinical studies, highlighting their abilities to inhibit the tumor proteasome and induce apoptosis in several human cancers.

  7. Identification of irradiated pepper by ESR measurement

    International Nuclear Information System (INIS)

    Morishita, Norio; Kume, Tamikazu; Kawakami, Waichiro; Ishigaki, Isao

    1988-01-01

    The possibility of using electron spin resonance (ESR) for identification of irradiated black and white pepper was examined. The ESR spectra induced by irradiation with 10 kGy are clearly distinguishable from that of unirradiated samples. The intensity of signal induced by irradiation was markedly reduced during 2 weeks storage but it was slowly reduced on further storage. The difference of intensity between the irradiated and unirradiated samples can be distinguished after 13 weeks storage. In this experiment, ground black pepper, ground white pepper and whole white pepper can be identified after 13 weeks. On the other hand, whole black pepper cannot be identified after 8 weeks because of the variation of values. It suggests that the suitable method for sample preparation such as separation of pellicle of whole black pepper is required before ESR measurement. (author)

  8. Mutation breeding in diffrent types of pepper

    International Nuclear Information System (INIS)

    2010-01-01

    This project was carried out under the collaboration of TAEK, SANAEM, and BATEM within 1999-2005 period. The aim of this project was to create new pepper varieties in Sera Demre 8 (green pepper) and ST59 (green pepper) cultivars which are important greenhouse cultivars by using mutation breeding methods. The Effective Mutagen Dose (ED50) was calculated by linear regression analyses. According to results, 166 Gy dose was found as ED50. At the end of the breeding cycle 14 new mutant lines were obtained from mutant population. These mutant lines are still using as genitor for F1 hybrid pepper breeding programs

  9. Pepper EST database: comprehensive in silico tool for analyzing the chili pepper (Capsicum annuum transcriptome

    Directory of Open Access Journals (Sweden)

    Kim Woo Taek

    2008-10-01

    Full Text Available Abstract Background There is no dedicated database available for Expressed Sequence Tags (EST of the chili pepper (Capsicum annuum, although the interest in a chili pepper EST database is increasing internationally due to the nutritional, economic, and pharmaceutical value of the plant. Recent advances in high-throughput sequencing of the ESTs of chili pepper cv. Bukang have produced hundreds of thousands of complementary DNA (cDNA sequences. Therefore, a chili pepper EST database was designed and constructed to enable comprehensive analysis of chili pepper gene expression in response to biotic and abiotic stresses. Results We built the Pepper EST database to mine the complexity of chili pepper ESTs. The database was built on 122,582 sequenced ESTs and 116,412 refined ESTs from 21 pepper EST libraries. The ESTs were clustered and assembled into virtual consensus cDNAs and the cDNAs were assigned to metabolic pathway, Gene Ontology (GO, and MIPS Functional Catalogue (FunCat. The Pepper EST database is designed to provide a workbench for (i identifying unigenes in pepper plants, (ii analyzing expression patterns in different developmental tissues and under conditions of stress, and (iii comparing the ESTs with those of other members of the Solanaceae family. The Pepper EST database is freely available at http://genepool.kribb.re.kr/pepper/. Conclusion The Pepper EST database is expected to provide a high-quality resource, which will contribute to gaining a systemic understanding of plant diseases and facilitate genetics-based population studies. The database is also expected to contribute to analysis of gene synteny as part of the chili pepper sequencing project by mapping ESTs to the genome.

  10. Visualizing Capsaicinoids: Colorimetric Analysis of Chili Peppers

    Science.gov (United States)

    Thompson, Robert Q.; Chu, Christopher; Gent, Robin; Gould, Alexandra P.; Rios, Laura; Vertigan, Theresa M.

    2012-01-01

    A colorimetric method for total capsaicinoids in chili pepper ("Capsicum") fruit is described. The placental material of the pepper, containing 90% of the capsaicinoids, was physically separated from the colored materials in the pericarp and extracted twice with methanol, capturing 85% of the remaining capsaicinoids. The extract, evaporated and…

  11. The Nrf1 CNC-bZIP protein is regulated by the proteasome and activated by hypoxia.

    Science.gov (United States)

    Chepelev, Nikolai L; Bennitz, Joshua D; Huang, Ting; McBride, Skye; Willmore, William G

    2011-01-01

    Nrf1 (nuclear factor-erythroid 2 p45 subunit-related factor 1) is a transcription factor mediating cellular responses to xenobiotic and pro-oxidant stress. Nrf1 regulates the transcription of many stress-related genes through the electrophile response elements (EpREs) located in their promoter regions. Despite its potential importance in human health, the mechanisms controlling Nrf1 have not been addressed fully. We found that proteasomal inhibitors MG-132 and clasto-lactacystin-β-lactone stabilized the protein expression of full-length Nrf1 in both COS7 and WFF2002 cells. Concomitantly, proteasomal inhibition decreased the expression of a smaller, N-terminal Nrf1 fragment, with an approximate molecular weight of 23 kDa. The EpRE-luciferase reporter assays revealed that proteasomal inhibition markedly inhibited the Nrf1 transactivational activity. These results support earlier hypotheses that the 26 S proteasome processes Nrf1 into its active form by removing its inhibitory N-terminal domain anchoring Nrf1 to the endoplasmic reticulum. Immunoprecipitation demonstrated that Nrf1 is ubiquitinated and that proteasomal inhibition increased the degree of Nrf1 ubiquitination. Furthermore, Nrf1 protein had a half-life of approximately 5 hours in COS7 cells. In contrast, hypoxia (1% O(2)) significantly increased the luciferase reporter activity of exogenous Nrf1 protein, while decreasing the protein expression of p65, a shorter form of Nrf1, known to act as a repressor of EpRE-controlled gene expression. Finally, the protein phosphatase inhibitor okadaic acid activated Nrf1 reporter activity, while the latter was repressed by the PKC inhibitor staurosporine. Collectively, our data suggests that Nrf1 is controlled by several post-translational mechanisms, including ubiquitination, proteolytic processing and proteasomal-mediated degradation as well as by its phosphorylation status. © 2011 Chepelev et al.

  12. Method of detecting irradiated pepper

    International Nuclear Information System (INIS)

    Doumaru, Takaaki; Furuta, Masakazu; Katayama, Tadashi; Toratani, Hirokazu; Takeda, Atsuhiko

    1989-01-01

    Spices represented by pepper are generally contaminated by microorganisms, and for using them as foodstuffs, some sterilizing treatment is indispensable. However, heating is not suitable to spices, accordingly ethylene oxide gas sterilization has been inevitably carried out, but its carcinogenic property is a problem. Food irradiation is the technology for killing microorganisms and noxious insects which cause the rotting and spoiling of foods and preventing the germination, which is an energy-conserving method without the fear of residual chemicals, therefore, it is most suitable to the sterilization of spices. In the irradiation of lower than 10 kGy, the toxicity test is not required for any food, and the irradiation of spices is permitted in 20 countries. However, in order to establish the international distribution organization for irradiated foods, the PR to consumers and the development of the means of detecting irradiation are the important subjects. The authors used pepper, and examined whether the hydrogen generated by irradiation remains in seeds and it can be detected or not. The experimental method and the results are reported. From the samples without irradiation, hydrogen was scarcely detected. The quantity of hydrogen generated was proportional to dose. The measuring instrument is only a gas chromatograph. (K.I.)

  13. Trial Watch: Proteasomal inhibitors for anticancer therapy.

    Science.gov (United States)

    Obrist, Florine; Manic, Gwenola; Kroemer, Guido; Vitale, Ilio; Galluzzi, Lorenzo

    2015-01-01

    The so-called "ubiquitin-proteasome system" (UPS) is a multicomponent molecular apparatus that catalyzes the covalent attachment of several copies of the small protein ubiquitin to other proteins that are generally (but not always) destined to proteasomal degradation. This enzymatic cascade is crucial for the maintenance of intracellular protein homeostasis (both in physiological conditions and in the course of adaptive stress responses), and regulates a wide array of signaling pathways. In line with this notion, defects in the UPS have been associated with aging as well as with several pathological conditions including cardiac, neurodegenerative, and neoplastic disorders. As transformed cells often experience a constant state of stress (as a result of the hyperactivation of oncogenic signaling pathways and/or adverse microenvironmental conditions), their survival and proliferation are highly dependent on the integrity of the UPS. This rationale has driven an intense wave of preclinical and clinical investigation culminating in 2003 with the approval of the proteasomal inhibitor bortezomib by the US Food and Drug Administration for use in multiple myeloma patients. Another proteasomal inhibitor, carfilzomib, is now licensed by international regulatory agencies for use in multiple myeloma patients, and the approved indications for bortezomib have been extended to mantle cell lymphoma. This said, the clinical activity of bortezomib and carfilzomib is often limited by off-target effects, innate/acquired resistance, and the absence of validated predictive biomarkers. Moreover, the antineoplastic activity of proteasome inhibitors against solid tumors is poor. In this Trial Watch we discuss the contribution of the UPS to oncogenesis and tumor progression and summarize the design and/or results of recent clinical studies evaluating the therapeutic profile of proteasome inhibitors in cancer patients.

  14. BAG3 induces the sequestration of proteasomal clients into cytoplasmic puncta : Implications for a proteasome-to-autophagy switch

    NARCIS (Netherlands)

    Minoia, Melania; Boncoraglio, Alessandra; Vinet, Jonathan; Morelli, Federica F.; Brunsting, Jeanette F.; Poletti, Angelo; Krom, Sabine; Reits, Eric; Kampinga, Harm H.; Carra, Serena

    Eukaryotic cells use autophagy and the ubiquitin-proteasome system as their major protein degradation pathways. Upon proteasomal impairment, cells switch to autophagy to ensure proper clearance of clients (the proteasome-to-autophagy switch). The HSPA8 and HSPA1A cochaperone BAG3 has been suggested

  15. BAG3 induces the sequestration of proteasomal clients into cytoplasmic puncta: implications for a proteasome-to-autophagy switch

    NARCIS (Netherlands)

    Minoia, Melania; Boncoraglio, Alessandra; Vinet, Jonathan; Morelli, Federica F.; Brunsting, Jeanette F.; Poletti, Angelo; Krom, Sabine; Reits, Eric; Kampinga, Harm H.; Carra, Serena

    2014-01-01

    Eukaryotic cells use autophagy and the ubiquitin-proteasome system as their major protein degradation pathways. Upon proteasomal impairment, cells switch to autophagy to ensure proper clearance of clients (the proteasome-to-autophagy switch). The HSPA8 and HSPA1A cochaperone BAG3 has been suggested

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

    Science.gov (United States)

    Schipper-Krom, Sabine; Juenemann, Katrin; Jansen, Anne H; Wiemhoefer, Anne; van den Nieuwendijk, Rianne; Smith, Donna L; Hink, Mark A; Bates, Gillian P; Overkleeft, Hermen; Ovaa, Huib; Reits, Eric

    2014-01-03

    Neurodegenerative disorders such as Huntington's disease are hallmarked by neuronal intracellular inclusion body formation. Whether proteasomes are irreversibly recruited into inclusion bodies in these protein misfolding disorders is a controversial subject. In addition, it has been proposed that the proteasomes may become clogged by the aggregated protein fragments, leading to impairment of the ubiquitin-proteasome system. Here, we show by fluorescence pulse-chase experiments in living cells that proteasomes are dynamically and reversibly recruited into inclusion bodies. As these recruited proteasomes remain catalytically active and accessible to substrates, our results challenge the concept of proteasome sequestration and impairment in Huntington's disease, and support the reported absence of proteasome impairment in mouse models of Huntington's disease. Copyright © 2013 Federation of European Biochemical Societies. All rights reserved.

  17. Bacterial proteasome activator bpa (rv3780) is a novel ring-shaped interactor of the mycobacterial proteasome.

    Science.gov (United States)

    Delley, Cyrille L; Laederach, Juerg; Ziemski, Michal; Bolten, Marcel; Boehringer, Daniel; Weber-Ban, Eilika

    2014-01-01

    The occurrence of the proteasome in bacteria is limited to the phylum of actinobacteria, where it is maintained in parallel to the usual bacterial compartmentalizing proteases. The role it plays in these organisms is still not fully understood, but in the human pathogen Mycobacterium tuberculosis (Mtb) the proteasome supports persistence in the host. In complex with the ring-shaped ATPase Mpa (called ARC in other actinobacteria), the proteasome can degrade proteins that have been post-translationally modified with the prokaryotic ubiquitin-like protein Pup. Unlike for the eukaryotic proteasome core particle, no other bacterial proteasome interactors have been identified to date. Here we describe and characterize a novel bacterial proteasome activator of Mycobacterium tuberculosis we termed Bpa (Rv3780), using a combination of biochemical and biophysical methods. Bpa features a canonical C-terminal proteasome interaction motif referred to as the HbYX motif, and its orthologs are only found in those actinobacteria encoding the proteasomal subunits. Bpa can inhibit degradation of Pup-tagged substrates in vitro by competing with Mpa for association with the proteasome. Using negative-stain electron microscopy, we show that Bpa forms a ring-shaped homooligomer that can bind coaxially to the face of the proteasome cylinder. Interestingly, Bpa can stimulate the proteasomal degradation of the model substrate β-casein, which suggests it could play a role in the removal of non-native or damaged proteins.

  18. Proteolytic Activity of the 26S Proteasome is required for the Meiotic Resumption, Germinal Vesicle Breakdown and Cumulus Expansion of Porcine Cumulus-Oocyte Complexes Matured In Vitro

    Czech Academy of Sciences Publication Activity Database

    Yi, Y. J.; Nagyová, Eva; Manandhar, G.; Procházka, Radek; Šutovsky, M.; Park, C. S.; Šutovský, P.

    2008-01-01

    Roč. 78, - (2008), s. 115-126 ISSN 0006-3363 R&D Projects: GA ČR GA305/05/0960 Institutional research plan: CEZ:AV0Z50450515 Keywords : cumulus expansion * FSH * germinal vesicle breakdown Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.469, year: 2008

  19. Targeting proteasomes in infectious organisms to combat disease.

    Science.gov (United States)

    Bibo-Verdugo, Betsaida; Jiang, Zhenze; Caffrey, Conor R; O'Donoghue, Anthony J

    2017-05-01

    Proteasomes are multisubunit, energy-dependent, proteolytic complexes that play an essential role in intracellular protein turnover. They are present in eukaryotes, archaea, and in some actinobacteria species. Inhibition of proteasome activity has emerged as a powerful strategy for anticancer therapy and three drugs have been approved for treatment of multiple myeloma. These compounds react covalently with a threonine residue located in the active site of a proteasome subunit to block protein degradation. Proteasomes in pathogenic organisms such as Mycobacterium tuberculosis and Plasmodium falciparum also have a nucleophilic threonine residue in the proteasome active site and are therefore sensitive to these anticancer drugs. This review summarizes efforts to validate the proteasome in pathogenic organisms as a therapeutic target. We describe several strategies that have been used to develop inhibitors with increased potency and selectivity for the pathogen proteasome relative to the human proteasome. In addition, we highlight a cell-based chemical screening approach that identified a potent, allosteric inhibitor of proteasomes found in Leishmania and Trypanosoma species. Finally, we discuss the development of proteasome inhibitors as anti-infective agents. © 2017 Federation of European Biochemical Societies.

  20. Study on some chemical changes in irradiated pepper and parsley

    International Nuclear Information System (INIS)

    Josimovic, L.

    1983-01-01

    The effect of ionizing radiation on some water soluble components (carbohydrates, carbonyl compounds) of pepper and parsley, as well as on piperine in pepper and chlorophyll in parsley has been investigated. The irradiation of parsley with doses as high as 5 Mrad does not bring about any distinct qualitative and quantitative changes. In irradiated pepper qualitative changes were also not observed, but quantitative changes were measured even for doses lower than 1 Mrad. However, piperine in pepper is quite resistant against radiation. (author)

  1. Study on some chemical changes in irradiated pepper and parsley

    Energy Technology Data Exchange (ETDEWEB)

    Josimovic, L [Institut za Nuklearne Nauke Boris Kidric, Belgrade (Yugoslavia)

    1983-05-01

    The effect of ionizing radiation on some water soluble components (carbohydrates, carbonyl compounds) of pepper and parsley, as well as on piperine in pepper and chlorophyll in parsley has been investigated. The irradiation of parsley with doses as high as 5 Mrad does not bring about any distinct qualitative and quantitative changes. In irradiated pepper qualitative changes were also not observed, but quantitative changes were measured even for doses lower than 1 Mrad. However, piperine in pepper is quite resistant against radiation.

  2. -pumping pyrophosphatase in pepper plants

    KAUST Repository

    Vigani, Gianpiero; Rolli, Eleonora; Marasco, Ramona; Dell'Orto, Marta; Michoud, Gregoire; Soussi, Asma; Raddadi, Noura; Borin, Sara; Sorlini, Claudia; Zocchi, Graziano; Daffonchio, Daniele

    2018-01-01

    It has been previously shown that the transgenic overexpression of the plant root vacuolar proton pumps H+ -ATPase (V-ATPase) and H+ -PPase (V-PPase) confer tolerance to drought. Since plant-root endophytic bacteria can also promote drought tolerance, we hypothesize that such promotion can be associated to the enhancement of the host vacuolar proton pumps expression and activity. To test this hypothesis, we selected two endophytic bacteria endowed with an array of in vitro plant growth promoting traits. Their genome sequences confirmed the presence of traits previously shown to confer drought resistance to plants, such as the synthesis of nitric oxide and of organic volatile organic compounds. We used the two strains on pepper (Capsicuum annuum L.) because of its high sensitivity to drought. Under drought conditions, both strains stimulated a larger root system and enhanced the leaves' photosynthetic activity. By testing the expression and activity of the vacuolar proton pumps, H+ -ATPase (V-ATPase) and H+ -PPase (V-PPase), we found that bacterial colonization enhanced V-PPase only. We conclude that the enhanced expression and activity of V-PPase can be favoured by the colonization of drought-tolerance-inducing bacterial endophytes. This article is protected by copyright. All rights reserved.

  3. -pumping pyrophosphatase in pepper plants

    KAUST Repository

    Vigani, Gianpiero

    2018-05-22

    It has been previously shown that the transgenic overexpression of the plant root vacuolar proton pumps H+ -ATPase (V-ATPase) and H+ -PPase (V-PPase) confer tolerance to drought. Since plant-root endophytic bacteria can also promote drought tolerance, we hypothesize that such promotion can be associated to the enhancement of the host vacuolar proton pumps expression and activity. To test this hypothesis, we selected two endophytic bacteria endowed with an array of in vitro plant growth promoting traits. Their genome sequences confirmed the presence of traits previously shown to confer drought resistance to plants, such as the synthesis of nitric oxide and of organic volatile organic compounds. We used the two strains on pepper (Capsicuum annuum L.) because of its high sensitivity to drought. Under drought conditions, both strains stimulated a larger root system and enhanced the leaves\\' photosynthetic activity. By testing the expression and activity of the vacuolar proton pumps, H+ -ATPase (V-ATPase) and H+ -PPase (V-PPase), we found that bacterial colonization enhanced V-PPase only. We conclude that the enhanced expression and activity of V-PPase can be favoured by the colonization of drought-tolerance-inducing bacterial endophytes. This article is protected by copyright. All rights reserved.

  4. Analysis of gamma irradiated pepper constituents, 5

    International Nuclear Information System (INIS)

    Takagi, Kazuko; Okuyama, Tsuneo; Ishikawa, Toshihiro.

    1988-01-01

    Gamma irradiated peppers (10 krad, 100 krad, 1 Mrad) were analyzed by HPLC. The extraction method and HPLC conditions were same as the first report, that is, the extraction from pepper was performed by Automatic Air Hammer and the extracted samples were separated on a reversed phase C 8 column with a concave gradient from 0.1% trifluoro aceticacid (TFA) in water to 75% acetonitrile-0.1% TFA in water for 60 minutes and detected at 210 nm, 280 nm. It is difficult to compare with irradiated and unirradiated pepper constituents by their peak height or area. And the method of multi variant statistically analysis was introduced. The 'peak n area/peak n + 1 area' ratio was calculated by computer. Each peak area was accounted by integrator. The value of these ratio were called 'parameter'. Each chromatogram has 741 parameters calculated with 39 chromatographic peaks. And these parameters were abopted to the multi variant statiscally analysis. Comparison of constituents between irradiated pepper and unirradiated pepper was done by 741 parameters. The correlation of parameters between irradiated and unirradiated was investigated by use of computer. Some parameters of irradiated case were selected as which had no correlation with unirradiated case. That is to say these parameters were thought to be changed with gamma spectrum irradiation. By this method, Coumarin was identified as a changed component with gamma irradiation. (author)

  5. The ubiquitin-proteasome system and autophagy are defective in the taurine-deficient heart.

    Science.gov (United States)

    Jong, Chian Ju; Ito, Takashi; Schaffer, Stephen W

    2015-12-01

    Taurine depletion leads to impaired mitochondrial function, as characterized by reduced ATP production and elevated superoxide generation. These defects can fundamentally alter cardiomyocyte function and if left unchanged can result in cell death. To protect against these stresses, cardiomyocytes possess quality control processes, such as the ubiquitin-proteasome system (UPS) and autophagy, which can rejuvenate cells through the degradation of damaged proteins and organelles. Hence, the present study tested the hypothesis that reactive oxygen species generated by damaged mitochondria initiates UPS and autophagy in the taurine-deficient heart. Using transgenic mice lacking the taurine transporter (TauTKO) as a model of taurine deficiency, it was shown that the levels of ubiquitinated protein were elevated, an effect associated with a decrease in ATP-dependent 26S β5 proteasome activity. Treating the TauTKO mouse with the mitochondria-specific antioxidant, mitoTEMPO, largely abolished the increase in ubiquitinated protein content. The TauTKO heart was also associated with impaired autophagy, characterized by an increase in the initiator, Beclin-1, and autophagosome content, but a defect in the generation of active autophagolysosomes. Although mitoTEMPO treatment only restores the oxidative balance within the mitochondria, it appeared to completely disrupt the crosstalk between the damaged mitochondria and the quality control processes. Thus, mitochondrial oxidative stress is the main trigger initiating the quality control systems in the taurine-deficient heart. We conclude that the activation of the UPS and autophagy is another fundamental function of mitochondria.

  6. Plant Virus Infection and the Ubiquitin Proteasome Machinery: Arms Race along the Endoplasmic Reticulum.

    Science.gov (United States)

    Verchot, Jeanmarie

    2016-11-19

    The endoplasmic reticulum (ER) is central to plant virus replication, translation, maturation, and egress. Ubiquitin modification of ER associated cellular and viral proteins, alongside the actions of the 26S proteasome, are vital for the regulation of infection. Viruses can arrogate ER associated ubiquitination as well as cytosolic ubiquitin ligases with the purpose of directing the ubiquitin proteasome system (UPS) to new targets. Such targets include necessary modification of viral proteins which may stabilize certain complexes, or modification of Argonaute to suppress gene silencing. The UPS machinery also contributes to the regulation of effector triggered immunity pattern recognition receptor immunity. Combining the results of unrelated studies, many positive strand RNA plant viruses appear to interact with cytosolic Ub-ligases to provide novel avenues for controlling the deleterious consequences of disease. Viral interactions with the UPS serve to regulate virus infection in a manner that promotes replication and movement, but also modulates the levels of RNA accumulation to ensure successful biotrophic interactions. In other instances, the UPS plays a central role in cellular immunity. These opposing roles are made evident by contrasting studies where knockout mutations in the UPS can either hamper viruses or lead to more aggressive diseases. Understanding how viruses manipulate ER associated post-translational machineries to better manage virus-host interactions will provide new targets for crop improvement.

  7. Plant Virus Infection and the Ubiquitin Proteasome Machinery: Arms Race along the Endoplasmic Reticulum

    Directory of Open Access Journals (Sweden)

    Jeanmarie Verchot

    2016-11-01

    Full Text Available The endoplasmic reticulum (ER is central to plant virus replication, translation, maturation, and egress. Ubiquitin modification of ER associated cellular and viral proteins, alongside the actions of the 26S proteasome, are vital for the regulation of infection. Viruses can arrogate ER associated ubiquitination as well as cytosolic ubiquitin ligases with the purpose of directing the ubiquitin proteasome system (UPS to new targets. Such targets include necessary modification of viral proteins which may stabilize certain complexes, or modification of Argonaute to suppress gene silencing. The UPS machinery also contributes to the regulation of effector triggered immunity pattern recognition receptor immunity. Combining the results of unrelated studies, many positive strand RNA plant viruses appear to interact with cytosolic Ub-ligases to provide novel avenues for controlling the deleterious consequences of disease. Viral interactions with the UPS serve to regulate virus infection in a manner that promotes replication and movement, but also modulates the levels of RNA accumulation to ensure successful biotrophic interactions. In other instances, the UPS plays a central role in cellular immunity. These opposing roles are made evident by contrasting studies where knockout mutations in the UPS can either hamper viruses or lead to more aggressive diseases. Understanding how viruses manipulate ER associated post-translational machineries to better manage virus–host interactions will provide new targets for crop improvement.

  8. Copper and manganese content of the leaves of pepper ( Capsicum ...

    African Journals Online (AJOL)

    The degree of Cu uptake by pepper plants was highest in the treatment on chernozem, lower on fluvisol and pseudogley, and lowest on vertisol, while the degree of Mn uptake by pepper plants was highest on chernozem and lowest on pseudogley. The Cu content of pepper leaves in all the treatments was low as compared ...

  9. 7 CFR 319.56-31 - Peppers from Spain.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Peppers from Spain. 319.56-31 Section 319.56-31... from Spain. Peppers (fruit) (Capsicum spp.) may be imported into the United States from Spain only... subpart: (a) The peppers must be grown in the Alicante or Almeria Province of Spain in pest-proof...

  10. ESR analysis of irradiated red peppers and commercial red peppers in Japan

    International Nuclear Information System (INIS)

    Kameya, Hiromi; Ukai, Mitsuko

    2011-01-01

    ESR analysis of γ-ray irradiated and being treated with different processing red pepper was studied. All the red peppers were commercial expect irradiated one. Processing treatment of red pepper was sun drying, mechanical processing (heating sterilization and powdering treatment). All the samples were weighted and analyzed. The ESR spectrum of the red pepper is composed of a singlet at g=2.00. This signal was originated from organic free radical. It is suggested the effect of heating treatment on the radical formation is not so large and powdering treatment will promote the radical formation of red pepper. ESR singlet signal of the irradiated red pepper showed the large signal intensity and the dose-dependence. The singlet signal intensity of irradiated powder sample showed the almost same value as compared with that of the powder sample with heating treatment. Relaxation times (T 1 and T 2 ) of the singlet signal were calculated. The relaxation behavior and relaxation times of the irradiated sample were different from that of the non-irradiated sample. The value of T 1 , the spin lattice relaxation time, of irradiated sample was increased and T 2 , the spin-spin relaxation time, of irradiated sample was decreased. We concluded that the radical formation of the red pepper is mainly depended on the powdering treatment and irradiation. (author)

  11. Involvement of proteasomal subunits zeta and iota in RNA degradation.

    Science.gov (United States)

    Petit, F; Jarrousse, A S; Dahlmann, B; Sobek, A; Hendil, K B; Buri, J; Briand, Y; Schmid, H P

    1997-01-01

    We have identified two distinct subunits of 20 S proteasomes that are associated with RNase activity. Proteasome subunits zeta and iota, eluted from two-dimensional Western blots, hydrolysed tobacco mosaic virus RNA, whereas none of the other subunits degraded this substrate under the same conditions. Additionally, proteasomes were dissociated by 6 M urea, and subunit zeta, containing the highest RNase activity, was isolated by anion-exchange chromatography and gel filtration. Purified subunit zeta migrated as a single spot on two-dimensional PAGE with a molecular mass of approx. 28 kDa. Addition of anti-(subunit zeta) antibodies led to the co-precipitation of this proteasome subunit and nuclease activity. This is the first evidence that proteasomal alpha-type subunits are associated with an enzymic activity, and our results provide further evidence that proteasomes may be involved in cellular RNA metabolism. PMID:9337855

  12. Effects of Commonly Used Pesticides in China on the Mitochondria and Ubiquitin-Proteasome System in Parkinson’s Disease

    Directory of Open Access Journals (Sweden)

    Tingting Chen

    2017-11-01

    Full Text Available Evidence continues to accumulate that pesticides are the leading candidates of environmental toxins that may contribute to the pathogenesis of Parkinson’s disease. The mechanisms, however, remain largely unclear. According to epidemiological studies, we selected nine representative pesticides (paraquat, rotenone, chlorpyrifos, pendimethalin, endosulfan, fenpyroximate, tebufenpyrad, trichlorphon and carbaryl which are commonly used in China and detected the effects of the pesticides on mitochondria and ubiquitin-proteasome system (UPS function. Our results reveal that all the nine studied pesticides induce morphological changes of mitochondria at low concentrations. Paraquat, rotenone, chlorpyrifos, pendimethalin, endosulfan, fenpyroximate and tebufenpyrad induced mitochondria fragmentation. Furthermore, some of them (paraquat, rotenone, chlorpyrifos, fenpyroximate and tebufenpyrad caused a significant dose-dependent decrease of intracellular ATP. Interestingly, these pesticides which induce mitochondria dysfunction also inhibit 26S and 20S proteasome activity. However, two out of the nine pesticides, namely trichlorphon and carbaryl, were found not to cause mitochondrial fragmentation or functional damage, nor inhibit the activity of the proteasome, which provides significant guidance for selection of pesticides in China. Moreover, our results demonstrate a potential link between inhibition of mitochondria and the UPS, and pesticide-induced Parkinsonism.

  13. Myostatin Activates the Ubiquitin-Proteasome and Autophagy-Lysosome Systems Contributing to Muscle Wasting in Chronic Kidney Disease

    Science.gov (United States)

    Wang, Dong-Tao; Yang, Ya-Jun; Huang, Ren-Hua; Zhang, Zhi-Hua; Lin, Xin

    2015-01-01

    Our evidence demonstrated that CKD upregulated the expression of myostatin, TNF-α, and p-IkBa and downregulated the phosphorylation of PI3K, Akt, and FoxO3a, which were also associated with protein degradation and muscle atrophy. The autophagosome formation and protein expression of autophagy-related genes were increased in muscle of CKD rats. The mRNA level and protein expression of MAFbx and MuRF-1 were also upregulated in CKD rats, as well as proteasome activity of 26S. Moreover, activation of myostatin elicited by TNF-α induces C2C12 myotube atrophy via upregulating the expression of autophagy-related genes, including MAFbx and MuRF1 and proteasome subunits. Inactivation of FoxO3a triggered by PI3K inhibitor LY294002 prevented the myostatin-induced increase of expression of MuRF1, MAFbx, and LC3-II protein in C2C12 myotubes. The findings were further consolidated by using siRNA interference and overexpression of myostatin. Additionally, expression of myostatin was activated by TNF-α via a NF-κB dependent pathway in C2C12 myotubes, while inhibition of NF-κB activity suppressed myostatin and improved myotube atrophy. Collectively, myostatin mediated CKD-induced muscle catabolism via coordinate activation of the autophagy and the ubiquitin-proteasome systems. PMID:26448817

  14. Proteasomal and Lysosomal Protein Degradation and Heart Disease

    OpenAIRE

    Wang, Xuejun; Robbins, Jeffrey

    2013-01-01

    In the cell, the proteasome and lysosomes represent the most important proteolytic machineries, responsible for the protein degradation in the ubiquitin-proteasome system (UPS) and autophagy, respectively. Both the UPS and autophagy are essential to protein quality and quantity control. Alterations in cardiac proteasomal and lysosomal degradation are remarkably associated with most heart disease in humans and are implicated in the pathogenesis of congestive heart failure. Studies carried out ...

  15. Targeting proteasomes in infectious organisms to combat disease

    OpenAIRE

    Bibo-Verdugo, B; Jiang, Z; Caffrey, CR; O'Donoghue, AJ

    2017-01-01

    Proteasomes are multisubunit, energy-dependent, proteolytic complexes that play an essential role in intracellular protein turnover. They are present in eukaryotes, archaea, and in some actinobacteria species. Inhibition of proteasome activity has emerged as a powerful strategy for anticancer therapy and three drugs have been approved for treatment of multiple myeloma. These compounds react covalently with a threonine residue located in the active site of a proteasome subunit to block protein...

  16. Hybrid breeding in pepper (capsicum annuum L.)

    International Nuclear Information System (INIS)

    Milkova, L.; Daskalov, S.

    1981-01-01

    Male sterile forms of peppers with gene sterility were produced by ν- and X-irradiation of dry seeds of cv. Pazarjishka kapiya 794 and Zlaten medal and subsequent selection in M 2 . Male sterile lines suitable for developing hybrid cultivars intended for various kinds of production were obtained by backcrossing with lines and cultivars having valuable economic characters and high combining ability. The promising pepper Belasitsa, Prista, Lyulin and Strouma (all of them for early field production), and Izoumroud and Prevuzhoden (for glasshouse production) are described. (authors)

  17. [Beliefs about chili pepper consumption and health in Mexico City].

    Science.gov (United States)

    López-Carrillo, L; Fernández-Ortega M, C; Costa-Dias, R; Franco-Marina, J; Alejandre-Badillo, T

    1995-01-01

    Eating chili peppers is a cultural tradition in Mexico. Controversial characteristics have been empirically associated to chili pepper consumption and human health. In this paper, the beliefs about the health impacts of chili pepper consumption in two independent groups of Mexico City residents are described. The results confirm, on the one hand, that there is a wide variety of health benefits and damages associated with chili pepper consumption, but on the other hand, that the levels of chili pepper consumption are not related to beliefs about its human health impact.

  18. PROSPECT OF INDONESIA BLACK AND WHITE PEPPERS EXPORT

    Directory of Open Access Journals (Sweden)

    Tavi Supriana

    2013-04-01

    Full Text Available AbstractThis study analyzes the behavior of pepper exports, consisting of black pepper and white pepper, to various destination countries. The results of this study showed that black pepper exports affected by the destination country's gross domestic product (GDP, the price of white pepper, the population of the country of destination and the exchange rate against the dollar. Meanwhile, white pepper exports affected by the destination country's GDP, the population of the country of destination, the price of black pepper, white pepper prices and the exchange rate against the dollar. The results also showed that black pepper and white pepper are not mutually substituted.Keywords: pepper, exports, GDP, population, exchange rate JEL Classificaiton Numbers: F14, F19AbstrakPenelitian ini menganalisis perilaku ekspor lada, terdiri dari lada hitam dan lada putih, ke berbagai negara tujuan. Hasil penelitian ini menunjukkan bahwa ekspor lada hitam dipengaruhi oleh Pendapatan domestik bruto (PDB negara tujuan, harga lada putih, populasi negara tujuan dan nilai tukar rupiah terhadap dolar. Sementara itu, ekspor lada putih dipengaruhi oleh PDB negara tujuan, populasi negara tujuan, harga lada hitam, harga lada putih dan nilai tukar rupiah terhadap dolar. Hasil penelitian juga menunjukkan bahwa lada hitam dan lada putih tidak saling bersubstitusi.Keywords: Lada, ekspor, PDB, populasi, kursJEL Classificaiton Numbers: F14, F19

  19. Structural Analysis of the Bacterial Proteasome Activator Bpa in Complex with the 20S Proteasome.

    Science.gov (United States)

    Bolten, Marcel; Delley, Cyrille L; Leibundgut, Marc; Boehringer, Daniel; Ban, Nenad; Weber-Ban, Eilika

    2016-12-06

    Mycobacterium tuberculosis harbors proteasomes that recruit substrates for degradation through an ubiquitin-like modification pathway. Recently, a non-ATPase activator termed Bpa (bacterial proteasome activator) was shown to support an alternate proteasomal degradation pathway. Here, we present the cryo-electron microscopy (cryo-EM) structure of Bpa in complex with the 20S core particle (CP). For docking into the cryo-EM density, we solved the X-ray structure of Bpa, showing that it forms tight four-helix bundles arranged into a 12-membered ring with a 40 Å wide central pore and the C-terminal helix of each protomer protruding from the ring. The Bpa model was fitted into the cryo-EM map of the Bpa-CP complex, revealing its architecture and striking symmetry mismatch. The Bpa-CP interface was resolved to 3.5 Å, showing the interactions between the C-terminal GQYL motif of Bpa and the proteasome α-rings. This docking mode is related to the one observed for eukaryotic activators with features specific to the bacterial complex. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Paradoxical resistance of multiple myeloma to proteasome inhibitors by decreased levels of 19S proteasomal subunits

    DEFF Research Database (Denmark)

    Acosta-Alvear, Diego; Cho, Min Y; Wild, Thomas

    2015-01-01

    Hallmarks of cancer, including rapid growth and aneuploidy, can result in non-oncogene addiction to the proteostasis network that can be exploited clinically. The defining example is the exquisite sensitivity of multiple myeloma (MM) to 20S proteasome inhibitors, such as carfilzomib. However, MM...

  1. Selective increase of in vivo firing frequencies in DA SN neurons after proteasome inhibition in the ventral midbrain.

    Science.gov (United States)

    Subramaniam, Mahalakshmi; Kern, Beatrice; Vogel, Simone; Klose, Verena; Schneider, Gaby; Roeper, Jochen

    2014-09-01

    The impairment of protein degradation via the ubiquitin-proteasome system (UPS) is present in sporadic Parkinson's disease (PD), and might play a key role in selective degeneration of vulnerable dopamine (DA) neurons in the substantia nigra pars compacta (SN). Further evidence for a causal role of dysfunctional UPS in familial PD comes from mutations in parkin, which results in a loss of function of an E3-ubiquitin-ligase. In a mouse model, genetic inactivation of an essential component of the 26S proteasome lead to widespread neuronal degeneration including DA midbrain neurons and the formation of alpha-synuclein-positive inclusion bodies, another hallmark of PD. Studies using pharmacological UPS inhibition in vivo had more mixed results, varying from extensive degeneration to no loss of DA SN neurons. However, it is currently unknown whether UPS impairment will affect the neurophysiological functions of DA midbrain neurons. To answer this question, we infused a selective proteasome inhibitor into the ventral midbrain in vivo and recorded single DA midbrain neurons 2 weeks after the proteasome challenge. We found a selective increase in the mean in vivo firing frequencies of identified DA SN neurons in anesthetized mice, while those in the ventral tegmental area (VTA) were unaffected. Our results demonstrate that a single-hit UPS inhibition is sufficient to induce a stable and selective hyperexcitability phenotype in surviving DA SN neurons in vivo. This might imply that UPS dysfunction sensitizes DA SN neurons by enhancing 'stressful pacemaking'. © 2014 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

  2. Analysis of gamma irradiated pepper constituents, 3

    International Nuclear Information System (INIS)

    Takagi, Kazuko; Okuyama, Tsuneo; Yamauchi, Yoshio; Saito, Muneo.

    1988-01-01

    The extraction of pepper perfume by use of supercritical fluid extraction (SFE) was investigated. Carbon dioxide was used as supercritical mobile phase. SFE was achieved by follow conditions, temperature was 40deg C, pressure was 200 kgf/cm 2 , 5 % methanol was added to mobile phase and the extraction time was 60 minutes. The extracted fraction by this method was yellow oily substance and was pepper perfume rich fraction, while the residue of extraction had not only perfume but also any pungency. And it seems that most part of pepper perfume was extracted by Supercritical fluid extraction. The perfume fraction was analyzed by two way method, that is, supercritical fluid chromatography (SFC) and reversed phase HPLC. SFC conditions were same as SFE. HPLC conditions were described as the first report. By both methods, about 7 peaks were detected. According to their analysis of this fraction, the main component was identified as piperine. On the chromatogram of reversed phase HPLC of extract, most peaks were eluted later than piperine. And it is thought that most perfume components have high hydrophobicity more than piperine. The change of perfume of pepper was able to be discussed based on the extraction by SFE. (author)

  3. Effect of alligator pepper (Zingiberaceae Aframomum melegueta ...

    African Journals Online (AJOL)

    Fifteen male rats and fifteen female rats of proven fertility from a pilot study were randomly paired in fifteen cages in a well ventillated room. After three days of mating, the males were withdrawn from ... risk to women in their first trimester of pregnancy. Key words : Effect, Alligator pepper, Herbal remedy, Nutrition, Pregnancy ...

  4. Impaired proteasome function in sporadic amyotrophic lateral sclerosis.

    Science.gov (United States)

    Kabashi, Edor; Agar, Jeffrey N; Strong, Michael J; Durham, Heather D

    2012-06-01

    Abstract The ubiquitin-proteasome system, important for maintaining protein quality control, is compromised in experimental models of familial ALS. The objective of this study was to determine if proteasome function is impaired in sporadic ALS. Proteasomal activities and subunit composition were evaluated in homogenates of spinal cord samples obtained at autopsy from sporadic ALS and non-neurological control cases, compared to cerebellum as a clinically spared tissue. The level of 20S α structural proteasome subunits was assessed in motor neurons by immunohistochemistry. Catalysis of peptide substrates of the three major proteasomal activities was substantially reduced in ALS thoracic spinal cord, but not in cerebellum, accompanied by alterations in the constitutive proteasome machinery. Chymotrypsin-like activity was decreased to 60% and 65% of control in ventral and dorsal spinal cord, respectively, concomitant with reduction in the β5 subunit with this catalytic activity. Caspase- and trypsin-like activities were reduced to a similar extent (46% - 68% of control). Proteasome levels, although generally maintained, appeared reduced specifically in motor neurons by immunolabelling. In conclusion, there are commonalities of findings in sporadic ALS patients and presymptomatic SOD1-G93A transgenic mice and these implicate inadequate proteasome function in the pathogenesis of both familial and sporadic ALS.

  5. Antitumorigenic effect of proteasome inhibitors on insulinoma cells

    DEFF Research Database (Denmark)

    Størling, Joachim; Allaman-Pillet, Nathalie; Karlsen, Allan E

    2004-01-01

    inhibition of the proteasome has an antitumorigenic potential in insulinoma cells. Exposure of mouse betaTC3 insulinoma cells to the proteasome inhibitor N-Acetyl-Leu-Leu-Nle-CHO (ALLN) reduced cell viability, activated caspase-3, induced apoptosis, and suppressed insulin release. Treatment with ALLN also...

  6. Detection of antibodies to the 20s proteasome by ELISA

    DEFF Research Database (Denmark)

    Jørgensen, Karin Meinike; Frederiksen, Jette Lautrup; Nielsen, Christoffer Tandrup

    2013-01-01

    The presence of antibodies against the 20S proteasome has been correlated with diseases like multiple sclerosis (MS) and systemic lupus erythematosus (SLE) but no definite association has been established. In order to investigate this further, we optimized an ELISA for proteasome antibodies...

  7. New Insights on Eggplant/Tomato/Pepper Synteny and Identification of Eggplant and Pepper Orthologous QTL

    Directory of Open Access Journals (Sweden)

    Riccardo Rinaldi

    2016-07-01

    Full Text Available Eggplant, pepper and tomato are the most exploited berry-producing vegetables within the Solanaceae family. Their genomes differ in size, but each has 12 chromosomes which have undergone rearrangements causing a redistribution of loci. The genome sequences of all three species are available but differ in coverage, assembly quality and percentage of anchorage.Determining their syntenic relationship and QTL orthology will contribute to exploit genomic resources and genetic data for key agronomic traits.The syntenic analysis between tomato and pepper based on the alignment of 34,727 tomato CDS to the pepper genome sequence, identified 19,734 unique hits. The resulting synteny map confirmed the 14 inversions and 10 translocations previously documented, but also highlighted 3 new translocations and 4 major new inversions. Furthermore, each of the 12 chromosomes exhibited a number of rearrangements involving small regions of 0.5-0.7 Mbp.Due to high fragmentation of the publicly available eggplant genome sequence, physical localization of most eggplant QTL was not possible, thus, we compared the organization of the eggplant genetic map with the genome sequence of both tomato and pepper. The eggplant/tomato syntenic map confirmed all the 10 translocations but only 9 of the 14 known inversions; on the other hand, a newly detected inversion was recognized while another one was not confirmed. The eggplant/pepper syntenic map confirmed 10 translocations and 8 inversions already detected and suggested a putative new translocation.In order to perform the assessment of eggplant and pepper QTL orthology, the eggplant and pepper sequence-based markers located in their respective genetic map were aligned onto the pepper genome. GBrowse in pepper was used as reference platform for QTL positioning. A set of 151 pepper QTL were located as well as 212 eggplant QTL, including 76 major QTL (PVE ≥ 10% affecting key agronomic traits. Most were confirmed to cluster in

  8. BAG3 induces the sequestration of proteasomal clients into cytoplasmic puncta: implications for a proteasome-to-autophagy switch.

    Science.gov (United States)

    Minoia, Melania; Boncoraglio, Alessandra; Vinet, Jonathan; Morelli, Federica F; Brunsting, Jeanette F; Poletti, Angelo; Krom, Sabine; Reits, Eric; Kampinga, Harm H; Carra, Serena

    2014-09-01

    Eukaryotic cells use autophagy and the ubiquitin-proteasome system as their major protein degradation pathways. Upon proteasomal impairment, cells switch to autophagy to ensure proper clearance of clients (the proteasome-to-autophagy switch). The HSPA8 and HSPA1A cochaperone BAG3 has been suggested to be involved in this switch. However, at present it is still unknown whether and to what extent BAG3 can indeed reroute proteasomal clients to the autophagosomal pathway. Here, we show that BAG3 induces the sequestration of ubiquitinated clients into cytoplasmic puncta colabeled with canonical autophagy linkers and markers. Following proteasome inhibition, BAG3 upregulation significantly contributes to the compensatory activation of autophagy and to the degradation of the (poly)ubiquitinated proteins. BAG3 binding to the ubiquitinated clients occurs through the BAG domain, in competition with BAG1, another BAG family member, that normally directs ubiquitinated clients to the proteasome. Therefore, we propose that following proteasome impairment, increasing the BAG3/BAG1 ratio ensures the "BAG-instructed proteasomal to autophagosomal switch and sorting" (BIPASS).

  9. New Sources of Tobamoviruses, CMV and Bacterial Spot Resistance in Pepper

    OpenAIRE

    Stoimenova, Elisaveta; Mitrev, Sasa; Bogatzevska, Nevena

    2005-01-01

    The pepper cultivars Zlaten medal, Alfi and Zalfi, the six Macedonian pepper accessions and the five Bulgarian lines have been screening for the resistance to cucumber mosaic virus (CMV), tobamoviruses and Xanthomonas vesicatoria pepper - tomato pathotype (XvPT).

  10. Genetic variability of Indonesian local chili pepper: The facts

    Science.gov (United States)

    Arumingtyas, Estri Laras; Kusnadi, Joni; Sari, Dewi Ratih Tirto; Ratih, Nursita

    2017-11-01

    Chili pepper (Capsicum frutescens) is one species of Solanaceae family that is very popular in Indonesia and some other tropical countries because of its pungency. Chili pepper is an important spice in Indonesia and is also eaten fresh as a pickle to increase appetite. In Indonesia, there are various local names for chili pepper includingcabai, cengek, lombok, pedesan etc. These varied local names represent the various morphological shapes of the chili pepper fruit. We have investigated the variability of some chili cultivars based on morphological characteristics, molecular markers, pungency, and capsaicin content. Some biological facts, such as the tendency of chili pepper to outbreed, have also been found. In this paper, the source of variability and the possible mechanism of increasing genetic variability of Indonesian local chili pepper are also discussed.

  11. Proteasome dynamics between proliferation and quiescence stages of Saccharomyces cerevisiae.

    Science.gov (United States)

    Yedidi, Ravikiran S; Fatehi, Amatullah K; Enenkel, Cordula

    The ubiquitin-proteasome system (UPS) plays a critical role in cellular protein homeostasis and is required for the turnover of short-lived and unwanted proteins, which are targeted by poly-ubiquitination for degradation. Proteasome is the key protease of UPS and consists of multiple subunits, which are organized into a catalytic core particle (CP) and a regulatory particle (RP). In Saccharomyces cerevisiae, proteasome holo-enzymes are engaged in degrading poly-ubiquitinated substrates and are mostly localized in the nucleus during cell proliferation. While in quiescence, the RP and CP are sequestered into motile and reversible storage granules in the cytoplasm, called proteasome storage granules (PSGs). The reversible nature of PSGs allows the proteasomes to be transported back into the nucleus upon exit from quiescence. Nuclear import of RP and CP through nuclear pores occurs via the canonical pathway that includes the importin-αβ heterodimer and takes advantage of the Ran-GTP gradient across the nuclear membrane. Dependent on the growth stage, either inactive precursor complexes or mature holo-enzymes are imported into the nucleus. The present review discusses the dynamics of proteasomes including their assembly, nucleo-cytoplasmic transport during proliferation and the sequestration of proteasomes into PSGs during quiescence. [Formula: see text].

  12. Proteasomal and lysosomal protein degradation and heart disease.

    Science.gov (United States)

    Wang, Xuejun; Robbins, Jeffrey

    2014-06-01

    In the cell, the proteasome and lysosomes represent the most important proteolytic machineries, responsible for the protein degradation in the ubiquitin-proteasome system (UPS) and autophagy, respectively. Both the UPS and autophagy are essential to protein quality and quantity control. Alterations in cardiac proteasomal and lysosomal degradation are remarkably associated with most heart disease in humans and are implicated in the pathogenesis of congestive heart failure. Studies carried out in animal models and in cell culture have begun to establish both sufficiency and, in some cases, the necessity of proteasomal functional insufficiency or lysosomal insufficiency as a major pathogenic factor in the heart. This review article highlights some recent advances in the research into proteasome and lysosome protein degradation in relation to cardiac pathology and examines the emerging evidence for enhancing degradative capacities of the proteasome and/or lysosome as a new therapeutic strategy for heart disease. This article is part of a Special Issue entitled "Protein Quality Control, the Ubiquitin Proteasome System, and Autophagy". Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Brazilian Capsicum peppers: capsaicinoid content and antioxidant activity.

    Science.gov (United States)

    Bogusz, Stanislau; Libardi, Silvia H; Dias, Fernanda Fg; Coutinho, Janclei P; Bochi, Vivian C; Rodrigues, Daniele; Melo, Arlete Mt; Godoy, Helena T

    2018-01-01

    Capsicum peppers are known as a source of capsaicinoids, phenolic compounds and antioxidants. Brazilian Capsicum peppers are important spices used in foods worldwide. However, little information is available on the chemical composition and antioxidant activity of these peppers. Capsaicin, dihydrocapsaicin, total phenolic compounds and antioxidant activity were investigated in extracts of three Brazilian peppers: Capsicum frutescens, C. chinense and C. baccatum var. pendulum, in two different harvest years and at two ripening stages. The bioactive compound content was dependent on harvest year, and changes in the concentration profiles were found for capsaicin. Mature fruits of C. chinense harvested in the first year had the highest capsaicin concentration (2.04 mg g -1 fresh pepper), and mature fruits of C. frutescens harvested in the same first year had the highest dihydrocapsaicin content (0.95 mg g -1 fresh pepper). Mature fruits of C. frutescens harvested in the first year showed the major total phenolic compound content (2.46 mg g -1 fresh pepper). The total phenolic compound content was directly related to antioxidant activity. Our results suggest that phenolic compounds significantly contribute to the antioxidant activity of the investigated peppers. Also, these data add valued novel information that enhances current knowledge of Brazilian pepper fruits. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  14. Analysis of gamma irradiated pepper constituents, 1

    International Nuclear Information System (INIS)

    Takagi, Kazuko; Okuyama, Tsuneo

    1988-01-01

    A reversed phase high performance liquid chromatographic (HPLC) method was developed for the analysis of many constituents of pepper at the same time. And a extraction method of ultraviolet absorbing constituents from pepper was developed for the HPLC analysis. The Ultraviolet absorbing constituents were extracted by precooled Automatic Air-Hammer from frozen pepper with 20% acetonitrile in water. The process of extraction was achieved under cooling by liquid nitrogen from start to end. The extracted constituents were separated on a reversed phase C 8 (LiChrospher 300 RP - 8 10 μm 0.4 I.D. x 0.4 cm and LiChrosorb RP - 8 SelectB 0.4 I. D. x 25 cm) column with a concave gradient from 0.1% trifluoro acetic acid (TFA) in water to 75% acetonitrile and 0.1% TFA in water for 60 minutes. The eluted constituents were detected 210 nm and 280 nm. The present method permits the detection of about 50 peaks by 280 nm. (author)

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

    NARCIS (Netherlands)

    Schipper-Krom, Sabine; Juenemann, Katrin; Jansen, Anne H.; Wiemhoefer, Anne; van den Nieuwendijk, Rianne; Smith, Donna L.; Hink, Mark A.; Bates, Gillian P.; Overkleeft, Hermen; Ovaa, Huib; Reits, Eric

    2014-01-01

    Neurodegenerative disorders such as Huntington's disease are hallmarked by neuronal intracellular inclusion body formation. Whether proteasomes are irreversibly recruited into inclusion bodies in these protein misfolding disorders is a controversial subject. In addition, it has been proposed that

  16. Changes in proteasome structure and function caused by HAMLET in tumor cells.

    Science.gov (United States)

    Gustafsson, Lotta; Aits, Sonja; Onnerfjord, Patrik; Trulsson, Maria; Storm, Petter; Svanborg, Catharina

    2009-01-01

    Proteasomes control the level of endogenous unfolded proteins by degrading them in the proteolytic core. Insufficient degradation due to altered protein structure or proteasome inhibition may trigger cell death. This study examined the proteasome response to HAMLET, a partially unfolded protein-lipid complex, which is internalized by tumor cells and triggers cell death. HAMLET bound directly to isolated 20S proteasomes in vitro and in tumor cells significant co-localization of HAMLET and 20S proteasomes was detected by confocal microscopy. This interaction was confirmed by co-immunoprecipitation from extracts of HAMLET-treated tumor cells. HAMLET resisted in vitro degradation by proteasomal enzymes and degradation by intact 20S proteasomes was slow compared to fatty acid-free, partially unfolded alpha-lactalbumin. After a brief activation, HAMLET inhibited proteasome activity in vitro and in parallel a change in proteasome structure occurred, with modifications of catalytic (beta1 and beta5) and structural subunits (alpha2, alpha3, alpha6 and beta3). Proteasome inhibition was confirmed in extracts from HAMLET-treated cells and there were indications of proteasome fragmentation in HAMLET-treated cells. The results suggest that internalized HAMLET is targeted to 20S proteasomes, that the complex resists degradation, inhibits proteasome activity and perturbs proteasome structure. We speculate that perturbations of proteasome structure might contribute to the cytotoxic effects of unfolded protein complexes that invade host cells.

  17. Studies on terrein as a new class of proteasome inhibitors

    International Nuclear Information System (INIS)

    Demasi, M.; Felicio, A.L.; Lima, C.; Pacheco, A.O.; Leite, H.G.; Andrade, L.H.

    2010-01-01

    The proteasome is an intracellular multicatalytic protease involved in the cell cycle regulation, signaling response, antigen presentation and apoptosis. Since proteasome inhibitors promote cell death by apoptosis, they have been proposed as new anti-tumoral drugs. Terrein, a secondary metabolite secreted by the fungus Aspergillus terreus, was firstly described in 1935. In the present work we report that terrein isolated through the screening for inhibitors of the 20S proteasome showed inhibitory effect upon both chymotrypsin- and trypsin-like activities of the multicatalytic core particle, the 20S proteasome. Despite of the high inhibitory concentration determined in vitro, that verified by incubating cells (fibroblasts and a pulmonary tumor cell line) in the presence of terrein was 4-fold lower indicating the proteasome as a selective intracellular target. Moreover, terrein promoted apoptotic cell death on both fibroblasts and pulmonary tumor cell line tested. Although terrein concentrations (mM range) necessary to elicit apoptosis in the cellular models herein tried were high when compared to those (muM and nM range) of other inhibitors recently described, its chemical structure is not correlated to any other inhibitor reported thus far. Therefore, the present results point out for the possibility of exploring terrein as a new molecular fragment for the development of synthetic proteasome inhibitors. (author)

  18. Sodium fluxes in sweet pepper exposed to varying sodium concentrations

    NARCIS (Netherlands)

    Blom-Zandstra, M.; Vogelzang, S.A.; Veen, B.W.

    1998-01-01

    The sodium transport and distribution of sweet pepper (Capsicum annuum L.) under saline conditions were studied after transferring the plants to a sodium-free nutrient solution. Sodium stress up to 60 mM did not affect the growth of sweet pepper, as it appears able to counteract the unfavourable

  19. Haematology and serum biochemistry of laying hens fed red pepper ...

    African Journals Online (AJOL)

    The hematology and serum biochemistry of ISA brown laying hens fed red pepper (Capsicum annum. L.) as feed additive in their diet was studied. Sixty (60) laying birds (in their 32nd week) were randomly allotted to four different dietary treatments with graded levels of red pepper (Capsicum annum. L.) as additive.

  20. Toward Valid Measurement of Stephen Pepper's World Hypotheses.

    Science.gov (United States)

    Johnson, John A.

    Two measures of the "world hypotheses" of Stephen Pepper were mailed to 100 sociobiologists, 87 behaviorists, 79 personality psychologists, and 45 human developmentalists. The World Hypothesis Scale (WHS) was designed to measure Pepper's four world views: (1) formism; (2) mechanism; (3) organicism; and (4) contextualism. The…

  1. Emittance formula for slits and pepper-pot measurement

    International Nuclear Information System (INIS)

    Zhang, M.

    1996-10-01

    In this note, a rigid formula for slits and pepper-pot emittance measurement is derived. The derivation is based on the one- dimensional slit measurement setup. A mathematical generalization of the slit emittance formula to the pepper-pot measurement is discussed

  2. Physicochemical properties of black pepper (Piper nigrum) starch.

    Science.gov (United States)

    Zhu, Fan; Mojel, Reuben; Li, Guantian

    2018-02-01

    Black pepper (Piper nigrum) is among the most popular spices around the world. Starch is the major component of black pepper. However, little is known about functional properties of this starch. In this study, swelling, solubility, thermal properties, rheology, and enzyme susceptibility of 2 black pepper starches were studied and compared with those of maize starch. Pepper starch had lower water solubility and swelling power than maize starch. It had higher viscosity during pasting event. In dynamic oscillatory analysis, pepper starch had lower storage modulus. Thermal analysis showed that pepper starch had much higher gelatinization temperatures (e.g., conclusion temperature of 94°C) than maize starch. The susceptibility to α-amylolysis of pepper starch was not very different from that of maize starch. Overall, the differences in the physicochemical properties of the 2 pepper starches are non-significant. The relationships between structure (especially amylopectin internal molecular structure) and properties of starch components are highlighted. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Coevolution of a Persistent Plant Virus and Its Pepper Hosts.

    Science.gov (United States)

    Safari, Maliheh; Roossinck, Marilyn J

    2018-05-30

    There are many nonpathogenic viruses that are maintained in a persistent lifestyle in plants. Plant persistent viruses are widespread, replicating in their hosts for many generations. So far, Endornaviridae is the only family of plant persistent viruses with a single-stranded RNA genome, containing one large open reading frame. Bell pepper endornavirus (BPEV), Hot pepper endornavirus, Capsicum frutescens endornavirus 1 (CFEV 1) have been identified from peppers. Peppers are native to Central and South America and, as domesticated plants, human selection accelerated their evolution. We investigated the evolution of these endornaviruses in different peppers including Capsicum annuum, C. chacoense, C.chinense, C. frutescens, C.bacccutum, and C. pubescens using two fragments from the viral helicase (Hel) and RNA dependent RNA polymerase (RdRp) domains. In addition, using single nucleotide polymorphisms, we analyzed the pepper host populations and phylogenies. The endornaviruses phylogeny was correlated with its Capsicum species host. In this study, BPEV was limited to C. annuum species, and the RdRp and Hel phylogenies identified two clades that correlated with the host pungency. No C. annuum infected with CFEV 1 was found in this study, but the CFEV 1 RdRp fragment was recovered from C. chinense, C. frutescens, C. bacccutum, and C. pubescens. Hence, during pepper speciation, the ancestor of CFEV 1 may have evolved as a new endornavirus, BPEV, in C. annuum peppers.

  4. Production of cell wall enzymes in pepper seedlings, inoculated with ...

    African Journals Online (AJOL)

    Pepper seedlings inoculated with arbuscular mycorrhizal AM fungus, Glomus etunicatum, produced cellulase, polygal-acturonase and pectin methylestrase enzymes. The activities of the enzymes increased as the pepper seedlings matured in age, showing that the activity of the enzymes in the seedlings was age mediated.

  5. Quantification of the gene expression of bell peppers ( Capsicum ...

    African Journals Online (AJOL)

    Fruits can be divided into two groups according to the regulatory mechanisms underlying their ripening process. The two ripening processes are climacteric and non-climacteric process; bell peppers are part of the non-climacteric plant groups. Bell peppers are members of the Solanacaea family. The Solanacaea family is ...

  6. Occurrence of pepper mild mottle virus in greenhouse- grown ...

    African Journals Online (AJOL)

    Administrator

    2011-06-08

    Jun 8, 2011 ... Pepper mild mottle virus (PMMoV) is one of the most ... Figure 1. Map of the West Mediterranean region of Turkey showing areas in which the surveys were conducted. showing virus-like symptoms were taken from symptomatic pepper .... SM, Maniloff J, Mayo MA, McGeoch D, Pringle CR, Wickner RB (eds).

  7. Serological detection of viruses infecting tomato and pepper in ...

    African Journals Online (AJOL)

    ... one tomato leaf sample while PVMV + CMV occurred on three pepper leaf samples. The control of aphid vectors that transmit these viruses and good sanitary practices against soil borne ToMV would minimize disease incidences and subsequent yield loss. Keywords: Tomato, Pepper, virus distribution, PVMV, CMV, PVY ...

  8. Nitrogen determination in pepper (Capsicum frutescens L.) plants by ...

    African Journals Online (AJOL)

    use

    2011-12-05

    Dec 5, 2011 ... significant negative relations between G/(R+G+B) ratio of coverage image and the indexes of pepper nitrogen status in this experiment. Hence, the suitable index of assessing pepper N status by color image analysis was discussed. MATERIALS AND METHODS. Experimental design. Experiment was made ...

  9. Identification of whitefly resistance in tomato and hot pepper

    NARCIS (Netherlands)

    Firdaus, S.

    2012-01-01

    Key words: Capsicum, Bemisia tabaci, trichome density, cuticle thickness
    Whitefly is economically one of the most threatening pests of pepper worldwide, which is mainly caused by its ability to transmit many different viruses. In this research, we characterized pepper germplasm to identify

  10. First report of "Candidatus Liberibacter solanacearum" on pepper in Honduras

    Science.gov (United States)

    In 2012, bell pepper (Capsicum annuum) plants exhibiting symptoms that resembled those of the bacterium “Candidatus Liberibacter solanacearum” infection were observed in commercial pepper fields in several departments in Honduras, including Francisco Morazán, Ocotepeque, El Paraíso, and Olancho. Man...

  11. Effect of aqueous extract of alligator pepper ( Aframomum malegueta )

    African Journals Online (AJOL)

    ... concentration 7days post intervention; and a significant decrease in Cl− concentration 14 days post-intervention. Our findings suggest that intraperitoneally injected aqueous extract of alligator pepper induces reversible changes in serum electrolytes. Key words: Alligator pepper, Electrolytes, Homeostasis, Renal function ...

  12. Paraquat, but not maneb, induces synucleinopathy and tauopathy in striata of mice through inhibition of proteasomal and autophagic pathways.

    Directory of Open Access Journals (Sweden)

    Jonathan Wills

    Full Text Available SNCA and MAPT genes and environmental factors are important risk factors of Parkinson's disease [PD], the second-most common neurodegenerative disease. The agrichemicals maneb and paraquat selectively target dopaminergic neurons, leading to parkinsonism, through ill-defined mechanisms. In the current studies we have analyzed the ability of maneb and paraquat, separately and together, to induce synucleinopathy and tauopathy in wild type mice. Maneb was ineffective in increasing α-synuclein [α-Syn] or p-Tau levels. By contrast, paraquat treatment of mice resulted in robust accumulation of α-Syn and hyperphosphorylation of Tau in striata, through activation of p-GSK-3β, a major Tau kinase. Co-treatment with maneb did not enhance the effects of paraquat. Increased hyperacetylation of α-tubulin was observed in paraquat-treated mice, suggesting cytoskeleton remodeling. Paraquat, but not maneb, inhibited soluble proteasomal activity on a peptide substrate but this was not associated with a decreased expression of 26S proteasome subunits. Both paraquat and maneb treatments increased levels of the autophagy inhibitor, mammalian target of rapamycin, mTOR, suggesting impaired axonal autophagy, despite increases in certain autophagic proteins, such as beclin 1 and Agt12. Autophagic flux was also impaired, as ratios of LC3 II to LC3 I were reduced in treated animals. Increased mTOR was also observed in postmortem human PD striata, where there was a reduction in the LC3 II to LC3 I ratio. Heat shock proteins were either increased or unchanged upon paraquat-treatment suggesting that chaperone-mediated autophagy is not hampered by the agrichemicals. These studies provide novel insight into the mechanisms of action of these agrichemicals, which indicate that paraquat is much more toxic than maneb, via its inhibitory effects on proteasomes and autophagy, which lead to accumulation of α-Syn and p-Tau.

  13. Investigations of the surviving microflora in irradiated black pepper

    International Nuclear Information System (INIS)

    Farkas, J.; Andrassy, E.

    1983-01-01

    The sensitizing effect of irradiation on the microflora of ground black pepper was studied with respect to heat, reduced pH-value, salt and nitrite, and the possible relationship between the sensitizing effects of irradiation and the water activity of the spice. The change of the sensitizing effect of irradiation after irradiation during storage of the black pepper was also studied at different storage temperatures and under various conditions of humidity. The effectivity of irradiation depends to a much lesser extent on the conditions of humidity than that of ethylene-oxide treatment. Following decontamination, the remaining microflora of irradiated black pepper would be more sensitive to antimicrobial influences predominating during its use in the food industry than that of ethylene-oxide treated black pepper. The increased sensitivity of the remaining microflora of the irradiated black pepper remains unchanged for at least 6 months. (author)

  14. Detection of gamma irradiated pepper and papain by chemiluminescence

    International Nuclear Information System (INIS)

    Sattar, Abdus; Delincee, H.; Diehl, J.F.

    1987-01-01

    Chemiluminescence (CL) measurements of black pepper and of papain using luminol and lucigenin reactions were studied. Effects of grinding, irradiation (5-20 kGy) and particle size (750-140 μm) on CL of pepper, and of irradiation (10-30 kGy) on CL of papain, were investigated. All the tested treatments affected the luminescence response in both the luminol and lucigenin reactions; however, the pattern of changes in each case, was inconsistent. Optimum pepper size for maximum luminescence was 560 μm, and optimum irradiation doses were > 15 kGy for pepper and > 20 kGy for papain. Chemiluminescence may possibly be used as an indicator or irradiation treatment for pepper and papain at a dose of 10 kGy or higher, but further research is needed to establish the reliability of this method. (author)

  15. On the role of proteasomes in cell biology and proteasome inhibition as a novel frontier in the development of immunosuppressants.

    Science.gov (United States)

    Wu, Jiangping

    2002-11-01

    The proteasome, a large protease complex in cells, is the major machinery for protein degradation. It was previously considered a humble garbage collector, performing housekeeping duties to remove misfolded or spent proteins. Until recently, the interests of immunologists in proteasomes were focused largely on its role in antigen processing. Its real importance in cell biology has only been revealed contemporarily due to the availability of relatively specific inhibitors. It has now become increasingly clear that many aspects of immune responses highly depend on proper proteasome activity. Recently, a proteasome inhibitor has been successfully used to prevent acute as well as ongoing heart allograft rejection in mice. Such inhibitors are also efficacious in treating several autoimmune diseases, such as arthritis, psoriasis, and probably type I diabetes, in animal models. Phase II and III clinical trials of proteasome inhibitors in treating various tumors have shown promising results, and the side-effects of these drugs are tolerable. Therefore, proteasome inhibition represents a new and promising frontier in immunosuppressant development.

  16. 7 CFR 457.148 - Fresh market pepper crop insurance provisions.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 6 2010-01-01 2010-01-01 false Fresh market pepper crop insurance provisions. 457.148... pepper crop insurance provisions. The fresh market pepper crop insurance provisions for the 1999 and... Fresh Market Pepper Crop Provisions If a conflict exists among the policy provisions, the order of...

  17. IMPROVED METHODS OF OBTAINING PEPPER SEEDLINGS

    OpenAIRE

    Florina Uleanu

    2012-01-01

    This paper refers to the effect of different types of pots on the level of growth and development of the pepper seedlings in order to clarify the influences caused by use of different recipes transplanters pots. Different biocomposites from renewable resources biodegradable nutritive support were studied. Seedlings were grown in 4 variants of pots M1 (V1), M2 (V2), M3 (V3) and jiffy- pots (V4). The height of the aerial part varied from 14.5 (V1) to 17.4 cm (V4), whereas the root length varied...

  18. Radiosensitizing effect of PSMC5, a 19S proteasome ATPase, in H460 lung cancer cells

    International Nuclear Information System (INIS)

    Yim, Ji-Hye; Yun, Hong Shik; Lee, Su-Jae; Baek, Jeong-Hwa; Lee, Chang-Woo; Song, Ji-Young; Um, Hong-Duck; Park, Jong Kuk; Kim, Jae-Sung; Park, In-Chul; Hwang, Sang-Gu

    2016-01-01

    The function of PSMC5 (proteasome 26S subunit, ATPase 5) in tumors, particularly with respect to cancer radioresistance, is not known. Here, we identified PSMC5 as a novel radiosensitivity biomarker, demonstrating that radiosensitive H460 cells were converted to a radioresistance phenotype by PSMC5 depletion. Exposure of H460 cells to radiation induced a marked accumulation of cell death-promoting reactive oxygen species, but this effect was blocked in radiation-treated H460 PSMC5-knockdown cells through downregulation of the p53-p21 pathway. Interestingly, PSMC5 depletion in H460 cells enhanced both AKT activation and MDM2 transcription, thereby promoting the degradation of p53 and p21 proteins. Furthermore, specific inhibition of AKT with triciribine or knockdown of MDM2 with small interfering RNA largely restored p21 expression in PSMC5-knockdown H460 cells. Our data suggest that PSMC5 facilitates the damaging effects of radiation in radiation-responsive H460 cancer cells and therefore may serve as a prognostic indicator for radiotherapy and molecular targeted therapy in lung cancer patients. - Highlights: • PSMC5 is a radiation-sensitive biomarker in H460 cells. • PSMC5 depletion inhibits radiation-induced apoptosis in H460 cells. • PSMC5 knockdown blocks ROS generation through inhibition of the p53-p21 pathway. • PSMC5 knockdown enhances p21 degradation via AKT-dependent MDM2 stabilization.

  19. Karyotypes, heterochromatin, and physical mapping of 18S-26S rDNA in Cactaceae.

    Science.gov (United States)

    Las Peñas, M L; Urdampilleta, J D; Bernardello, G; Forni-Martins, E R

    2009-01-01

    Karyotype analyses in members of the four Cactaceae subfamilies were performed. Numbers and karyotype formula obtained were: Pereskioideae = Pereskiaaculeata(2n = 22; 10 m + 1 sm), Maihuenioideae = Maihuenia patagonica (2n = 22, 9 m + 2 sm; 2n = 44, 18 m + 4 sm), Opuntioideae = Cumulopuntia recurvata(2n = 44; 20 m + 2 sm), Cactoideae = Acanthocalycium spiniflorum (2n = 22; 10 m + 1 sm),Echinopsis tubiflora (2n = 22; 10 m + 1 sm), Trichocereus candicans (2n = 22, 22 m). Chromosomes were small, the average chromosome length was 2.3 mum. Diploid species and the tetraploid C. recurvata had one terminal satellite, whereas the remaining tetraploid species showed four satellited chromosomes. Karyotypes were symmetrical. No CMA(-)/DAPI(+) bands were detected, but CMA(+)/DAPI(-) bands associated with NOR were always found. Pericentromeric heterochromatin was found in C. recurvata, A. spiniflorum, and the tetraploid cytotype of M. patagonica. The locations of the 18S-26S rDNA sites in all species coincided with CMA(+)/DAPI(-) bands; the same occurred with the sizes and numbers of signals for each species. This technique was applied for the first time in metaphase chromosomes in cacti. NOR-bearing pair no.1 may be homeologous in all species examined. In Cactaceae, the 18S-26S loci seem to be highly conserved. Copyright 2009 S. Karger AG, Basel.

  20. Proteasome Inhibition Suppresses Dengue Virus Egress in Antibody Dependent Infection.

    Directory of Open Access Journals (Sweden)

    Milly M Choy

    2015-11-01

    Full Text Available The mosquito-borne dengue virus (DENV is a cause of significant global health burden, with an estimated 390 million infections occurring annually. However, no licensed vaccine or specific antiviral treatment for dengue is available. DENV interacts with host cell factors to complete its life cycle although this virus-host interplay remains to be fully elucidated. Many studies have identified the ubiquitin proteasome pathway (UPP to be important for successful DENV production, but how the UPP contributes to DENV life cycle as host factors remains ill defined. We show here that proteasome inhibition decouples infectious virus production from viral RNA replication in antibody-dependent infection of THP-1 cells. Molecular and imaging analyses in β-lactone treated THP-1 cells suggest that proteasome function does not prevent virus assembly but rather DENV egress. Intriguingly, the licensed proteasome inhibitor, bortezomib, is able to inhibit DENV titers at low nanomolar drug concentrations for different strains of all four serotypes of DENV in primary monocytes. Furthermore, bortezomib treatment of DENV-infected mice inhibited the spread of DENV in the spleen as well as the overall pathological changes. Our findings suggest that preventing DENV egress through proteasome inhibition could be a suitable therapeutic strategy against dengue.

  1. Association of proteasomal activity with metastasis in luminal breast cancer

    Science.gov (United States)

    Shashova, E. E.; Fesik, E. A.; Doroshenko, A. V.

    2017-09-01

    Chimotrypsin-like (ChTL) and caspase-like (CL) proteasomal activities were investigated in different variants of the tumor progression of luminal breast cancer. Patients with primary luminal breast cancer (n = 123) in stage T1-3N0-2M0 who had not received neoadjuvant treatment were included in this study. Proteasome ChTL and CL activities were determined in the samples of tumor and adjacent tissues. The coefficients of chymotrypsin-like (kChTL) and caspase-like (kCL) proteasome activity were also calculated as the ratio of the corresponding activity in the tumor tissue to activity in the adjacent tissue. ChTL, CL, kChTL and kCL in the tissues of luminal A and B breast cancer with lymphogenic metastasis were compared, and their association with hematogenous metastasis was evaluated. On the one hand, CL activity of proteasomes increased in luminal A breast cancer with extensive lymphogenic metastasis (N2), on the other hand it decreased in the luminal B subtype of cancer. The ratio of proteasomal activity in the tumor and adjacent tissues plays a significant role in the hematogenic pathway of breast cancer progression and is associated with poor metastatic-free survival.

  2. Irradiated black pepper identification based on thermoluminescence of silicate minerals

    International Nuclear Information System (INIS)

    Faycal Kharfi; Randa Ketfi

    2018-01-01

    In this work we have successfully implemented thermoluminescence TL method for irradiated food identification. First tests are performed on Indian black pepper and show promising results to extend the proposed method to many other foods. The X-ray diffraction (XRD) shows that SiO 2 (p3 2 21) is the main component of the separated mineral phase. A saturation dose of ∼ 100 Gy is determined for this pepper above which all thermoluminescent centers of the quartz are activated. Thus, above this threshold dose, only the pepper irradiation will be confirmed but no accuracy on the exact dose received. (author)

  3. Conditions of viscosity measurement for detecting irradiated peppers

    International Nuclear Information System (INIS)

    Hayashi, Toru; Todoriki, Setsuko; Okadome, Hiroshi; Kohyama, Kaoru

    1995-01-01

    Viscosity of gelatinized suspensions of black and white peppers decreased depending upon dose. The viscosity was influenced by gelatinization and viscosity measurement conditions. The difference between unirradiated pepper and an irradiated one was larger at a higher pH and temperature for gelatinization. A viscosity parameter normalized with the starch content of pepper sample and the viscosity of a 5% suspension of corn starch could get rid of the influence of the conditions for viscosity measurement such as type of viscometer, shear rate and temperature. (author)

  4. Irradiated pepper and ginger detected by viscosity and starch measurements

    International Nuclear Information System (INIS)

    Wegmueler, F.; Meier, W.

    1999-01-01

    Starch and rheological measurements of alkaline suspensions of white pepper, black pepper and ginger are a useful tool to distinguish not irradiated samples from the gamma-treated spices (dose gtoreq 2 kGy). In addition it is shown that starch is not the material which determines the different rheological behaviour of the alkaline suspensions of the spices. The differences in the viscosity data are rather due to irradiation damages of polymers which are enriched in the cell wall material of the pepper grains and the roots of the ginger

  5. SPR imaging biosensor for the 20S proteasome: Sensor development and application to measurement of proteasomes in human blood plasma

    International Nuclear Information System (INIS)

    Gorodkiewicz, E.; Sankiewicz, A.; Ostrowska, H.

    2011-01-01

    The 20S proteasome is a multicatalytic enzyme complex responsible for intracellular protein degradation in mammalian cells. Its antigen level or enzymatic activity in blood plasma are potentially useful markers for various malignant and nonmalignant diseases. We have developed a method for highly selective determination of the 20S proteasome using a Surface Plasmon Resonance Imaging (SPRI) technique. It is based on the highly selective interaction between the proteasome's catalytic β5 subunit and immobilized inhibitors (the synthetic peptide PSI and epoxomicin). Inhibitor concentration and pH were optimized. Analytical responses, linear ranges, accuracy, precision and interferences were investigated. Biosensors based on either PSI and epoxomicin were found to be suitable for quantitative determination of the proteasome, with a precision of ±10% for each, and recoveries of 102% and 113%, respectively, and with little interference by albumin, trypsin, chymotrypsin, cathepsin B and papain. The proteasome also was determined in plasma of healthy subjects and of patients suffering from acute leukemia. Both biosensors gave comparable results (2860 ng.mL -1 on average for control, and 42300 ng.mL -1 on average for leukemia patients). (author)

  6. Transcriptional upregulation of BAG3 upon proteasome inhibition

    International Nuclear Information System (INIS)

    Wang Huaqin; Liu Haimei; Zhang Haiyan; Guan Yifu; Du Zhenxian

    2008-01-01

    Proteasome inhibitors exhibit antitumoral activity against malignancies of different histology. Emerging evidence indicates that antiapoptotic factors may also accumulate as a consequence of exposure to these drugs, thus it seems plausible that activation of survival signaling cascades might compromise their antitumoral effects. Bcl-2-associated athanogene (BAG) family proteins are characterized by their property of interaction with a variety of partners involved in modulating the proliferation/death balance, including heat shock proteins (HSP), Bcl-2, Raf-1. In this report, we demonstrated that BAG3 is a novel antiapoptotic molecule induced by proteasome inhibitors in various cancer cells at the transcriptional level. Moreover, we demonstrated that BAG3 knockdown by siRNA sensitized cancer cells to MG132-induced apoptosis. Taken together, our results suggest that BAG3 induction might represents as an unwanted molecular consequence of utilizing proteasome inhibitors to combat tumors

  7. Local and systemic hormonal responses in pepper leaves during compatible and incompatible pepper-tobamovirus interactions

    Czech Academy of Sciences Publication Activity Database

    Dziurka, M.; Janeczko, A.; Juhasz, C.; Gullner, G.; Oklešťková, Jana; Novák, Ondřej; Saja, D.; Skoczowski, A.; Tobias, I.; Barna, B.

    2016-01-01

    Roč. 109, DEC (2016), s. 355-364 ISSN 0981-9428 R&D Projects: GA ČR GA14-34792S Institutional support: RVO:61389030 Keywords : tobacco-mosaic-virus * pathogenesis-related proteins * salicylic-acid * abscisic-acid * acquired- resistance * disease resistance * nicotiana-benthamiana * arabidopsis-thaliana * defense response * immune-responses * Brassinosteroids * Ethylene * Hormone * Pepper * Phenylalanine ammonia lyase * Progesterone * Salicylic acid * Tobamovirus Subject RIV: EF - Botanics Impact factor: 2.724, year: 2016

  8. The use of activity based protein profiling to study proteasome biology

    NARCIS (Netherlands)

    Paniagua Soriano, Guillem

    2016-01-01

    The work described in this thesis focuses on the characterization of proteasome directed activity-based probes (ABPs) as well as on the adaptation mechanisms that make multiple myeloma derived cell lines resistant against proteasome inhibitors (PIs).

  9. Discovery and development of inhibitors selective for human constitutive proteasome and immunoproteasome active sites

    NARCIS (Netherlands)

    Xin, B.

    2017-01-01

    This thesis describes the design and development of subunit‐selective inhibitors of particular catalytically active subunits of human constitutive proteasomes and immunoproteasomes. Most existing proteasome inhibitors are oligopeptides composed of 2‐4 amino acid residues, N‐terminally

  10. Prediction of proteasome cleavage motifs by neural networks

    DEFF Research Database (Denmark)

    Kesimir, C.; Nussbaum, A.K.; Schild, H.

    2002-01-01

    physiological conditions. Our algorithm has been trained not only on in vitro data, but also on MHC Class I ligand data, which reflect a combination of immunoproteasome and constitutive proteasome specificity. This feature, together with the use of neural networks, a non-linear classification technique, make...... the prediction of MHC Class I ligand boundaries more accurate: 65% of the cleavage sites and 85% of the non-cleavage sites are correctly determined. Moreover, we show that the neural networks trained on the constitutive proteasome data learns a specificity that differs from that of the networks trained on MHC...

  11. Impaired methylation as a novel mechanism for proteasome suppression in liver cells

    Energy Technology Data Exchange (ETDEWEB)

    Osna, Natalia A., E-mail: nosna@UNMC.edu [Liver Study Unit, The Omaha Veterans Affairs VA Medical Center, Omaha, NE 68105 (United States); Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68105 (United States); White, Ronda L.; Donohue, Terrence M. [Liver Study Unit, The Omaha Veterans Affairs VA Medical Center, Omaha, NE 68105 (United States); Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68105 (United States); Beard, Michael R. [Department of Molecular Biosciences, University of Adelaide (Australia); Tuma, Dean J.; Kharbanda, Kusum K. [Liver Study Unit, The Omaha Veterans Affairs VA Medical Center, Omaha, NE 68105 (United States); Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68105 (United States)

    2010-01-08

    The proteasome is a multi-catalytic protein degradation enzyme that is regulated by ethanol-induced oxidative stress; such suppression is attributed to CYP2E1-generated metabolites. However, under certain conditions, it appears that in addition to oxidative stress, other mechanisms are also involved in proteasome regulation. This study investigated whether impaired protein methylation that occurs during exposure of liver cells to ethanol, may contribute to suppression of proteasome activity. We measured the chymotrypsin-like proteasome activity in Huh7CYP cells, hepatocytes, liver cytosols and nuclear extracts or purified 20S proteasome under conditions that maintain or prevent protein methylation. Reduction of proteasome activity of hepatoma cell and hepatocytes by ethanol or tubercidin was prevented by simultaneous treatment with S-adenosylmethionine (SAM). Moreover, the tubercidin-induced decline in proteasome activity occurred in both nuclear and cytosolic fractions. In vitro exposure of cell cytosolic fractions or highly purified 20S proteasome to low SAM:S-adenosylhomocysteine (SAH) ratios in the buffer also suppressed proteasome function, indicating that one or more methyltransferase(s) may be associated with proteasomal subunits. Immunoblotting a purified 20S rabbit red cell proteasome preparation using methyl lysine-specific antibodies revealed a 25 kDa proteasome subunit that showed positive reactivity with anti-methyl lysine. This reactivity was modified when 20S proteasome was exposed to differential SAM:SAH ratios. We conclude that impaired methylation of proteasome subunits suppressed proteasome activity in liver cells indicating an additional, yet novel mechanism of proteasome activity regulation by ethanol.

  12. Diagnostic of the beam by pepper-pot method

    CERN Document Server

    Dolinskaya, M E; Zajchenko, A K

    2002-01-01

    The new method for the measuring of the particle distribution in the transversal plane with pepper-pot device is described. The algorithm for the beam emittance and Twiss parameters determination, using such measurements is presented. 7 refs., 8 figs.

  13. Hot Chili Peppers: Extraction, Cleanup, and Measurement of Capsaicin

    Science.gov (United States)

    Huang, Jiping; Mabury, Scott A.; Sagebiel, John C.

    2000-12-01

    Capsaicin, the pungent ingredient of the red pepper or Capsicum annuum, is widely used in food preparation. The purpose of this experiment was to acquaint students with the active ingredients of hot chili pepper (capsaicin and dihydrocapsaicin), the extraction, cleanup, and analysis of these chemicals, as a fun and informative analytical exercise. Fresh peppers were prepared and extracted with acetonitrile, removing plant co-extractives by addition to a C-18 solid-phase extraction cartridge. Elution of the capsaicinoids was accomplished with a methanol-acetic acid solution. Analysis was completed by reverse-phase HPLC with diode-array or variable wavelength detection and calibration with external standards. Levels of capsaicin and dihydrocapsaicin were typically found to correlate with literature values for a specific hot pepper variety. Students particularly enjoyed relating concentrations of capsaicinoids to their perceived valuation of "hotness".

  14. Genetic diversity in some Turkish pepper (Capsicum annuum L ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-15

    Sep 15, 2009 ... RFLP, RAPD, AFLP, and micro-satellites have been beneficial by being ... List of pepper genotypes used in the experiment. Genotype. Type. Source. Classification of ..... Artificial selection might decrease the genetic diversity ...

  15. Identification of irradiated pepper with comet assay

    International Nuclear Information System (INIS)

    Prieto Miranda, Enrique Fco.; Moreno Alvarez, Damaris L.; Carro Palacio, Sandra; Iglesia Enriquez, Isora

    2007-01-01

    The treatment of foods with ionizing radiations is a technological process utilized in order to increase the hygienic quality and the storage time of the foods. Several methods of detection of irradiated foods have been recommended. The comet assay of DNA is one fast and economical technique for the qualitative identification of irradiated foods. The objective of the present paper was to identify with the comet assay technique the modifications of the DNA molecule of irradiated pepper storage at environment and refrigeration temperatures and different post-irradiation times for different absorbed dose values, (0.1, 0.3 and 0.5 kGy). It was demonstrated that for the high absorbed dose values was observed a greater break into fragments of the DNA molecule, which shows the application of this technique for the identification of irradiated foods. (author)

  16. Evidence for a complex of emergent poleroviruses affecting pepper worldwide.

    Science.gov (United States)

    Fiallo-Olivé, Elvira; Navas-Hermosilla, Elisa; Ferro, Camila G; Zerbini, F Murilo; Navas-Castillo, Jesús

    2018-05-01

    In recent years, symptoms of vein yellowing and leaf roll in pepper crops associated with the presence of poleroviruses (genus Polerovirus, family Luteoviridae) have been emerging in many countries worldwide. Spain was the first country in Europe where the yellowing disease of pepper was observed. In this work, a polerovirus isolate from Spain that infects pepper and has been shown to be transmitted by the aphid Aphis gossyppii (Spain-Almería 2-2013) was sequenced and compared with isolates from Japan, Israel, China and Australia. The genome (6125 nt in length, GenBank accession number KY523072) of the isolate from Spain has the typical organization of poleroviruses and contains seven open reading frames (ORF0 to ORF5 and ORF3a), putatively encoding proteins P0 to P5 and P3a. A comparison of the sequence from Spain with the four complete sequences available for poleroviruses infecting pepper showed a closer relationship to the isolate from Israel and supports the existence of a complex of at least five polerovirus species. Given that the symptoms caused by all pepper poleroviruses described to date are similar, if not identical, we propose to name them "pepper vein yellows virus 1" to "pepper vein yellows virus 5" (PeVYV-1 to PeVYV-5), with PeVYV-5 corresponding to the polerovirus from Spain described in this work. Our results and those published over the last few years have shown that the emergent poleroviruses threatening pepper crops around the world are highly complex due to recombination events.

  17. Drying hot red pepper using solar tunnel drier

    International Nuclear Information System (INIS)

    Hossain, M.A; Bala, B.K.

    2006-01-01

    A solar tunnel drier was used to dry red hot pepper under the tropical weather conditions of Bangladesh in order to investigate its performance and the quality of the drier product. The drier comprises a plastic sheet-covered flat plate collector and a drying tunnel. The drier is arranged to supply hot air to the drying tunnel using two small fans powered by a 40 watt PV module. Fresh red pepper was water blanched before drying. In each drying batch in the solar tunnel drier, 20 kg of dried red pepper with 4 to 6% moisture content (wb) was obtained from 80 kg of fresh red pepper with initial moisture content of 73 to 75% (wb) in 20 to 22 hours of drying while it took 32 to 34 hours to bring down the moisture content of similar sample to 8 to 10% (wb) in sun drying methods. The pepper dried in the solar tunnel drier was completely protected from dust, dirt, rain, insects, birds, rodents and microorganisms and it was a quality-dried product in term of colour and pungency. The solar tunnel drier is recommended for drying of pepper as well as vegetables and fruits in developing countries especially in Bangladesh

  18. Effect of Red Pepper ( Powder or Red Pepper Pigment on the Performance and Egg Yolk Color of Laying Hens

    Directory of Open Access Journals (Sweden)

    Huaqiang Li

    2012-11-01

    Full Text Available Two experiments were conducted to study the effects of red pepper (Capsicum frutescens powder or red pepper pigment on the performance and egg yolk color of laying hens. In Exp. 1, 210, thirty-wk old, Hy-line Brown laying hens were fed one of seven diets containing 0.3, 0.6, 1.2, 2.0, 4.8 or 9.6 ppm red pepper pigment or 0.3 ppm carophyll red. Each diet was fed to three replicate batteries of hens with each battery consisting of a row of five cages of hens with two hens per cage (n = 3. In Exp. 2, 180, thirty-wk old, Hyline Brown laying hens, housed similarly to those in Exp. 1, were fed an unsupplemented basal diet as well as treatments in which the basal diet was supplemented with 0.8% red pepper powder processed in a laboratory blender to an average particle size of 300 μm, 0.8% red pepper powder processed as a super fine powder with a vibrational mill (44 μm and finally 0.8% red pepper powder processed as a super fine powder with a vibrational mill but mixed with 5% Na2CO3 either before or after grinding. A diet supplemented with 0.3 ppm carophyll red pigment was also included (n = 3. In both experiments, hens were fed the red pepper powder or pigment for 14 days. After feeding of the powder or pigment was terminated, all hens were fed the basal diet for eight more days to determine if the dietary treatments had any residual effects. In Exp. 1, there were no differences in egg-laying performance, feed consumption or feed conversion ratio due to inclusion of red pepper pigment in the diet. Average egg weight was higher (p0.05. However, compared with the control group, supplementation with all of the red pepper powder treatments increased egg weight (p<0.05. All the red pepper powder treatments also increased (p<0.05 the yolk color score compared with the control. The results of the present study suggest that both red pepper powder and pigment are effective feed additives for improving egg yolk color for laying hens.

  19. Decontamination of Black Pepper (Pepper Negrum) in grains by irradiation. Descontaminacion de pimienta negra (Pipper Negrum) en grano por irradiacion

    Energy Technology Data Exchange (ETDEWEB)

    Alvarez, M; Pino, J; Zayas, M [Instituto de Investigaciones para la Industria Alimenticia, La Habana (Cuba)

    1991-01-01

    The effect of irradiation with doses ranging between 3 and 10 kGy on the microbiological quality, on different chemical components and on the organoleptic quality of black pepper in grains was studied in this paper. The results showed the effectiveness of treatment in the decontamination of this spice. Piperin content, yield of essential oil and its composition as well as the organoleptic quality did not change with applied doses to black pepper in grains.

  20. Predicting proteasomal cleavage sites: a comparison of available methods

    DEFF Research Database (Denmark)

    Saxova, P.; Buus, S.; Brunak, Søren

    2003-01-01

    -terminal, in particular, of CTL epitopes is cleaved precisely by the proteasome, whereas the N-terminal is produced with an extension, and later trimmed by peptidases in the cytoplasm and in the endoplasmic reticulum. Recently, three publicly available methods have been developed for prediction of the specificity...

  1. Functional interplay between mitochondrial and proteasome activity in skin aging

    NARCIS (Netherlands)

    Kozie, Rafa; Greussing, Ruth; Maier, Andrea B.; Declercq, Lieve; Jansen-Dürr, Pidder

    According to the mitochondrial theory of aging, reactive oxygen species (ROS) derived primarily from mitochondria cause cumulative oxidative damage to various cellular molecules and thereby contribute to the aging process. On the other hand, a pivotal role of the proteasome, as a main proteolytic

  2. Proteasome nuclear import mediated by Arc3 can influence efficient DNA damage repair and mitosis in Schizosaccharomyces pombe

    DEFF Research Database (Denmark)

    Cabrera, Rodrigo; Sha, Zhe; Vadakkan, Tegy J.

    2010-01-01

    Proteasomes must remove regulatory molecules and abnormal proteins throughout the cell, but how proteasomes can do so efficiently remains unclear. We have isolated a subunit of the Arp2/3 complex, Arc3, which binds proteasomes. When overexpressed, Arc3 rescues phenotypes associated with proteasom...

  3. Suppression of 19S proteasome subunits marks emergence of an altered cell state in diverse cancers.

    Science.gov (United States)

    Tsvetkov, Peter; Sokol, Ethan; Jin, Dexter; Brune, Zarina; Thiru, Prathapan; Ghandi, Mahmoud; Garraway, Levi A; Gupta, Piyush B; Santagata, Sandro; Whitesell, Luke; Lindquist, Susan

    2017-01-10

    The use of proteasome inhibitors to target cancer's dependence on altered protein homeostasis has been greatly limited by intrinsic and acquired resistance. Analyzing data from thousands of cancer lines and tumors, we find that those with suppressed expression of one or more 19S proteasome subunits show intrinsic proteasome inhibitor resistance. Moreover, such proteasome subunit suppression is associated with poor outcome in myeloma patients, where proteasome inhibitors are a mainstay of treatment. Beyond conferring resistance to proteasome inhibitors, proteasome subunit suppression also serves as a sentinel of a more global remodeling of the transcriptome. This remodeling produces a distinct gene signature and new vulnerabilities to the proapoptotic drug, ABT-263. This frequent, naturally arising imbalance in 19S regulatory complex composition is achieved through a variety of mechanisms, including DNA methylation, and marks the emergence of a heritably altered and therapeutically relevant state in diverse cancers.

  4. Black pepper and health claims: a comprehensive treatise.

    Science.gov (United States)

    Butt, Masood Sadiq; Pasha, Imran; Sultan, Muhammad Tauseef; Randhawa, Muhammad Atif; Saeed, Farhan; Ahmed, Waqas

    2013-01-01

    For millennia, spices have been an integral part of human diets and commerce. Recently, the widespread recognition of diet-health linkages bolsters their dietary importance. The bioactive components present in them are of considerable significance owing to their therapeutic potential against various ailments. They provide physiological benefits or prevent chronic ailment in addition to the fundamental nutrition and often included in the category of functional foods. Black pepper (Piper Nigrum L.) is an important healthy food owing to its antioxidant, antimicrobial potential and gastro-protective modules. Black pepper, with piperine as an active ingredient, holds rich phytochemistry that also includes volatile oil, oleoresins, and alkaloids. More recently, cell-culture studies and animal modeling predicted the role of black pepper against number of maladies. The free-radical scavenging activity of black pepper and its active ingredients might be helpful in chemoprevention and controlling progression of tumor growth. Additionally, the key alkaloid components of Piper Nigrum, that is, piperine assist in cognitive brain functioning, boost nutrient's absorption and improve gastrointestinal functionality. In this comprehensive treatise, efforts are made to elucidate the antioxidant, antimicrobial, anti-inflammatory, gastro-protective, and antidepressant activities of black pepper. Moreover, the synergistic interaction of black pepper with different drugs and nutrients is the limelight of the manuscript. However, the aforementioned health-promoting benefits associated with black pepper are proven in animal modeling. Thus, there is a need to conduct controlled randomized trials in human subjects, cohort studies, and meta-analyses. Such future studies would be helpful in recommending its application in diet-based regimens to prevent various ailments.

  5. Susceptibility of ornamental pepper banker plant candidates to common greenhouse pests

    Science.gov (United States)

    Susceptibility of four potential ornamental pepper banker plant candidates [Black Pearl (BP), Explosive Ember (EE), Masquerade (MA), Red Missile (RM), and a commercial pepper cultivar Blitz (BL)] were evaluated against three common greenhouse pests - Bemisia tabaci, Polyphagotarsonemus latus and Fra...

  6. Technological innovation of induced breeding of space and selection of new variety on pepper

    International Nuclear Information System (INIS)

    Guo Yahua; Xie Libo; Wang Xue; Deng Liping

    2004-01-01

    New pepper variety Yujiao No.1 and new pepper lines with different horticultural characters are obtained by new breeding technique: combination of space inducement and regular breeding, field selection and inherited marker

  7. Research of photoinduced potentials of pepper leaves

    Directory of Open Access Journals (Sweden)

    D. V. Chernetchenko

    2015-09-01

    Full Text Available Bioelectrical potentials of the plants during photostimulation with various intensity values and wave lengths are studied. Suchstimulation provides light-dependent reactions of photosynthesis. The universal scheme of registration of bioelectric potentials, which allowsto carry out the experimental study of light stimuli effect on the plant is proposed. All registrations of potentials are observed on the pepper leaves. Photoinduced stimuli are different with intensity changed from 150 to 450 lux and different spectrums of white light and red light with wave length 690 nm. Different parameters of stimuli drive to different types of biopotentials of the plant, quantitative and qualitative relationships of this process are obtained. It is also found that bioelectrical reaction of the leaves is quantitatively different in various spectral ranges of light, but the quality remains the same for all cases, as shown in the study of bioelectric responses during exposure to the light in red spectrum range. The explanation of the additional depolarization phase that occurs during the running of dark reparative process of photosynthesis is given. Light-dependent reactions of photosynthesis are accompanied by significant changes in the electrical potential of chloroplasts’ membrane. Besides, the dependence of bioelectrical reaction of the plant on different spectra of light is found. By this way, biophysical relation of plant potentials to the intracellular biophysical mechanisms during photo stimulation is obtained.

  8. IMPROVED METHODS OF OBTAINING PEPPER SEEDLINGS

    Directory of Open Access Journals (Sweden)

    Florina Uleanu

    2012-12-01

    Full Text Available This paper refers to the effect of different types of pots on the level of growth and development of the pepper seedlings in order to clarify the influences caused by use of different recipes transplanters pots. Different biocomposites from renewable resources biodegradable nutritive support were studied. Seedlings were grown in 4 variants of pots M1 (V1, M2 (V2, M3 (V3 and jiffy- pots (V4. The height of the aerial part varied from 14.5 (V1 to 17.4 cm (V4, whereas the root length varied from 5.4 (V1 to 12.6 cm (V4. The number of leaves ranged from 7 (V2 to 12 (V4. The total volume was lowest for V2 (1.5 cm3 and highest for V4 (2.5 cm3, but the root volume was lowest for V2 (0.5 cm3 and highest for V1 (1 cm3. V2 also resulted in the smallest total seedling mass (1.6 g aerial part mass (1.1 g and root mass (0.5 g. Excepting the root volume V4 had the greatest values for the studied indicators.

  9. Bioactivities of anastasia black (Russian sweet pepper).

    Science.gov (United States)

    Shirataki, Yoshiaki; Kawase, Masami; Sakagami, Hiroshi; Nakashima, Hideki; Tani, Satoru; Tanaka, Toru; Sohara, Yoshitaka; Schelz, Zsuzsanna; Molnar, Joseph; Motohashi, Noboru

    2005-01-01

    Anastasia Black (Russian sweet pepper) of Capsicum annuum L. var. angulosum Mill. (Solanaceae) was successively extracted with hexane, acetone, methanol and 70% methanol, and the extracts were further separated into a total of twenty-three fractions by silica gel or octadecylsilane (ODS; C18) column chromatography. These extracts and fractions were investigated for their cytotoxicity, anti-human immunodeficiency virus (HIV), anti-Helicobacter pylori (H. pylori), urease inhibition and multidrug resistance (MDR) reversal activity. Some fractions of hexane and acetone extracts showed higher cytotoxic activity against three human oral tumor cell lines (squamous cell carcinoma HSC-2, HSC-3, submandibular gland carcinoma HSG) than against three normal human oral cells (gingival fibroblast HGF, pulp cell HPC, periodontal ligament fibroblast HPLF), suggesting a tumor-specific cytotoxic activity. No fractions displayed anti-HIV activity, but some hydrophobic fractions showed higher anti-H. pylori activity, urease inhibition activity and MDR reversal activity. The higher MDR activity of these fractions against MDR gene-transfected L5178 mouse lymphoma T cells may possibly be due to their higher content of carotene or polyphenol. These data suggest that Anastasia Black should be further investigated as a potent supplement for cancer chemotherapy.

  10. Pungency Quantitation of Hot Pepper Sauces Using HPLC

    Science.gov (United States)

    Betts, Thomas A.

    1999-02-01

    A class of compounds known as capsaicinoids are responsible for the "heat" of hot peppers. To determine the pungency of a particular pepper or pepper product, one may quantify the capsaicinoids and relate those concentrations to the perceived heat. The format of the laboratory described here allows students to collectively develop an HPLC method for the quantitation of the two predominant capsaicinoids (capsaicin and dihydrocapsaicin) in hot-pepper products. Each small group of students investigated one of the following aspects of the method: detector wavelength, mobile-phase composition, extraction of capsaicinoids, calibration, and quantitation. The format of the lab forced students to communicate and cooperate to develop this method. The resulting HPLC method involves extraction with acetonitrile followed by solid-phase extraction clean-up, an isocratic 80:20 methanol-water mobile phase, a 4.6 mm by 25 cm C-18 column, and UV absorbance detection at 284 nm. The method developed by the students was then applied to the quantitation of capsaicinoids in a variety of hot pepper sauces. Editor's Note on Hazards in our April 2000 issue addresses the above.

  11. Garlic exerts allelopathic effects on pepper physiology in a hydroponic co-culture system

    Directory of Open Access Journals (Sweden)

    Haiyan Ding

    2016-05-01

    Full Text Available A hydroponic co-culture system was adopted to determine the allelopathic potential of garlic on the growth of pepper plants. Different numbers of garlic plants (0, 2, 4, 8 and 12 were hydroponically co-cultured with two pepper plants to investigate allelopathic effects on the growth attributes and antioxidative defense system of the test pepper plants. The responses of the pepper plants depended on the number of garlic plants included in the co-culture system, indicating an association of pepper growth with the garlic root exudate concentration. When grown at a pepper/garlic ratio of 1:1 or 1:2, the pepper plant height, chlorophyll content, and peroxidase (POD, catalase (CAT and phenylalanine ammonia-lyase (PAL activities were significantly increased after 30 days of co-culture; in contrast, reduction in methane dicarboxylic aldehyde (MDA content was observed. However, when the pepper/garlic ratio was 1:4 or higher, these morphological indices and protective enzyme activities were significantly inhibited, whereas MDA levels in the pepper leaves were significantly increased due to severe membrane lipid peroxidation. The results indicate that although low concentrations of garlic root exudates appear to induce protective enzyme systems and promote pepper growth, high concentrations have deleterious effects. These findings suggest that further investigations should optimize the co-culture pepper/garlic ratio to reduce continuous cropping obstacles in pepper production.

  12. Role of the Ubiquitin-Proteasome Systems in the Biology and Virulence of Protozoan Parasites

    Directory of Open Access Journals (Sweden)

    Christian Muñoz

    2015-01-01

    Full Text Available In eukaryotic cells, proteasomes perform crucial roles in many cellular pathways by degrading proteins to enforce quality control and regulate many cellular processes such as cell cycle progression, signal transduction, cell death, immune responses, metabolism, protein-quality control, and development. The catalytic heart of these complexes, the 20S proteasome, is highly conserved in bacteria, yeast, and humans. However, until a few years ago, the role of proteasomes in parasite biology was completely unknown. Here, we summarize findings about the role of proteasomes in protozoan parasites biology and virulence. Several reports have confirmed the role of proteasomes in parasite biological processes such as cell differentiation, cell cycle, proliferation, and encystation. Proliferation and cell differentiation are key steps in host colonization. Considering the importance of proteasomes in both processes in many different parasites such as Trypanosoma, Leishmania, Toxoplasma, and Entamoeba, parasite proteasomes might serve as virulence factors. Several pieces of evidence strongly suggest that the ubiquitin-proteasome pathway is also a viable parasitic therapeutic target. Research in recent years has shown that the proteasome is a valid drug target for sleeping sickness and malaria. Then, proteasomes are a key organelle in parasite biology and virulence and appear to be an attractive new chemotherapeutic target.

  13. Effect of ionizing radiation exposure on Trypanosoma cruzi ubiquitin-proteasome system.

    Science.gov (United States)

    Cerqueira, Paula G; Passos-Silva, Danielle G; Vieira-da-Rocha, João P; Mendes, Isabela Cecilia; de Oliveira, Karla A; Oliveira, Camila F B; Vilela, Liza F F; Nagem, Ronaldo A P; Cardoso, Joseane; Nardelli, Sheila C; Krieger, Marco A; Franco, Glória R; Macedo, Andrea M; Pena, Sérgio D J; Schenkman, Sérgio; Gomes, Dawidson A; Guerra-Sá, Renata; Machado, Carlos R

    2017-03-01

    In recent years, proteasome involvement in the damage response induced by ionizing radiation (IR) became evident. However, whether proteasome plays a direct or indirect role in IR-induced damage response still unclear. Trypanosoma cruzi is a human parasite capable of remarkable high tolerance to IR, suggesting a highly efficient damage response system. Here, we investigate the role of T. cruzi proteasome in the damage response induced by IR. We exposed epimastigotes to high doses of gamma ray and we analyzed the expression and subcellular localization of several components of the ubiquitin-proteasome system. We show that proteasome inhibition increases IR-induced cell growth arrest and proteasome-mediated proteolysis is altered after parasite exposure. We observed nuclear accumulation of 19S and 20S proteasome subunits in response to IR treatments. Intriguingly, the dynamic of 19S particle nuclear accumulation was more similar to the dynamic observed for Rad51 nuclear translocation than the observed for 20S. In the other hand, 20S increase and nuclear translocation could be related with an increase of its regulator PA26 and high levels of proteasome-mediated proteolysis in vitro. The intersection between the opposed peaks of 19S and 20S protein levels was marked by nuclear accumulation of both 20S and 19S together with Ubiquitin, suggesting a role of ubiquitin-proteasome system in the nuclear protein turnover at the time. Our results revealed the importance of proteasome-mediated proteolysis in T. cruzi IR-induced damage response suggesting that proteasome is also involved in T. cruzi IR tolerance. Moreover, our data support the possible direct/signaling role of 19S in DNA damage repair. Based on these results, we speculate that spatial and temporal differences between the 19S particle and 20S proteasome controls proteasome multiple roles in IR damage response. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Role of the Ubiquitin Proteasome System in Regulating Skin Pigmentation

    Directory of Open Access Journals (Sweden)

    Hideya Ando

    2009-10-01

    Full Text Available Pigmentation of the skin, hair and eyes is regulated by tyrosinase, the critical rate-limiting enzyme in melanin synthesis by melanocytes. Tyrosinase is degraded endogenously, at least in part, by the ubiquitin proteasome system (UPS. Several types of inherited hypopigmentary diseases, such as oculocutaneous albinism and Hermansky-Pudlak syndrome, involve the aberrant processing and/or trafficking of tyrosinase and its subsequent degradation which can occur due to the quality-control machinery. Studies on carbohydrate modifications have revealed that tyrosinase in the endoplasmic reticulum (ER is proteolyzed via ER-associated protein degradation and that tyrosinase degradation can also occur following its complete maturation in the Golgi. Among intrinsic factors that regulate the UPS, fatty acids have been shown to modulate tyrosinase degradation in contrasting manners through increased or decreased amounts of ubiquitinated tyrosinase that leads to its accelerated or decelerated degradation by proteasomes.

  15. Safety assessment for genetically modified sweet pepper and tomato

    International Nuclear Information System (INIS)

    Chen Zhangliang; Gu Hongya; Li Yi; Su Yilan; Wu Ping; Jiang Zhicheng; Ming Xiaotian; Tian Jinhua; Pan Naisui; Qu Lijia

    2003-01-01

    The coat protein (CP) gene of cucumber mosaic virus (CMV) was cloned from a Chinese CMV isolate, the CaMV promoter and NOS terminator added and the gene construct was transformed into both sweet pepper and tomato plants to confer resistance to CMV. Safety assessments of these genetically modified (GM) plants were conducted. It was found that these two GM products showed no genotoxicity either in vitro or in vivo by the micronucleus test, sperm aberration test and Ames test. Animal feeding studies showed no significant differences in growth, body weight gain, food consumption, hematology, blood biochemical indices, organ weights and histopathology between rats or mice of either sex fed with either GM sweet pepper or tomato diets compared with those with non-GM diets. These results demonstrate that the CMV-resistant sweet pepper and tomato are comparable to the non-GM counterparts in terms of food safety

  16. Investigating radiostimulation effects of tomatoes, pepper and aubergines productivity improvement

    International Nuclear Information System (INIS)

    Daskalov, Khr.; Mal'tseva, S.

    1974-01-01

    The paper presents results of a study on the stimulating effect of ionizing radiation with low doses on tomatoes, pepper, and eggplants with a view to determining optimal radiation doses and possible practical applications of this method. The tomato varieties used were those most common in Bulgaria. It was found that (a) presowing irradiation of tomato, pepper, and eggplant seeds (graded by moisture content) with low doses of gamma rays from Co 60 had a strong stimulating effect; (b) radiation did not cause any changes in chemical composition or palatability; (c) irradiated variants produced more fruits per plant; and (d) the optimal doses are 1500-2500 r for tomatoes, 1200-1800 r for pepper, and 1500-3000 r for eggplants. (E.T.)

  17. Puromycin induces SUMO and ubiquitin redistribution upon proteasome inhibition

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, Hotaru [Course for Biological Sciences, Faculty of Science, Kumamoto University, Kumamoto (Japan); Saitoh, Hisato, E-mail: hisa@kumamoto-u.ac.jp [Course for Biological Sciences, Faculty of Science, Kumamoto University, Kumamoto (Japan); Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, Kumamoto (Japan)

    2016-07-29

    We have previously reported the co-localization of O-propargyl-puromycin (OP-Puro) with SUMO-2/3 and ubiquitin at promyelocytic leukemia-nuclear bodies (PML-NBs) in the presence of the proteasome inhibitor MG132, implying a role for the ubiquitin family in sequestering OP-puromycylated immature polypeptides to the nucleus during impaired proteasome activity. Here, we found that as expected puromycin induced SUMO-1/2/3 accumulation with ubiquitin at multiple nuclear foci in HeLa cells when co-exposed to MG132. Co-administration of puromycin and MG132 also facilitated redistribution of PML and the SUMO-targeted ubiquitin ligase RNF4 concurrently with SUMO-2/3. As removal of the drugs from the medium led to disappearance of the SUMO-2/3-ubiquitin nuclear foci, our findings indicated that nuclear assembly/disassembly of SUMO-2/3 and ubiquitin was pharmacologically manipulable, supporting our previous observation on OP-Puro, which predicted the ubiquitin family function in sequestrating aberrant proteins to the nucleus. -- Highlights: •Puromycin exhibits the O-propargyl-puromycin effect. •Puromycin induces SUMO redistribution upon proteasome inhibition. •Ubiquitin and RNF4 accumulate at PML-nuclear bodies with SUMO-2/3. •The ubiquitin family may function in nuclear sequestration of immature proteins.

  18. Puromycin induces SUMO and ubiquitin redistribution upon proteasome inhibition

    International Nuclear Information System (INIS)

    Matsumoto, Hotaru; Saitoh, Hisato

    2016-01-01

    We have previously reported the co-localization of O-propargyl-puromycin (OP-Puro) with SUMO-2/3 and ubiquitin at promyelocytic leukemia-nuclear bodies (PML-NBs) in the presence of the proteasome inhibitor MG132, implying a role for the ubiquitin family in sequestering OP-puromycylated immature polypeptides to the nucleus during impaired proteasome activity. Here, we found that as expected puromycin induced SUMO-1/2/3 accumulation with ubiquitin at multiple nuclear foci in HeLa cells when co-exposed to MG132. Co-administration of puromycin and MG132 also facilitated redistribution of PML and the SUMO-targeted ubiquitin ligase RNF4 concurrently with SUMO-2/3. As removal of the drugs from the medium led to disappearance of the SUMO-2/3-ubiquitin nuclear foci, our findings indicated that nuclear assembly/disassembly of SUMO-2/3 and ubiquitin was pharmacologically manipulable, supporting our previous observation on OP-Puro, which predicted the ubiquitin family function in sequestrating aberrant proteins to the nucleus. -- Highlights: •Puromycin exhibits the O-propargyl-puromycin effect. •Puromycin induces SUMO redistribution upon proteasome inhibition. •Ubiquitin and RNF4 accumulate at PML-nuclear bodies with SUMO-2/3. •The ubiquitin family may function in nuclear sequestration of immature proteins.

  19. Discovery of an Inhibitor of the Proteasome Subunit Rpn11.

    Science.gov (United States)

    Perez, Christian; Li, Jing; Parlati, Francesco; Rouffet, Matthieu; Ma, Yuyong; Mackinnon, Andrew L; Chou, Tsui-Fen; Deshaies, Raymond J; Cohen, Seth M

    2017-02-23

    The proteasome plays a crucial role in degradation of normal proteins that happen to be constitutively or inducibly unstable, and in this capacity it plays a regulatory role. Additionally, it degrades abnormal/damaged/mutant/misfolded proteins, which serves a quality-control function. Inhibitors of the proteasome have been validated in the treatment of multiple myeloma, with several FDA-approved therapeutics. Rpn11 is a Zn 2+ -dependent metalloisopeptidase that hydrolyzes ubiquitin from tagged proteins that are trafficked to the proteasome for degradation. A fragment-based drug discovery (FBDD) approach was utilized to identify fragments with activity against Rpn11. Screening of a library of metal-binding pharmacophores (MBPs) revealed that 8-thioquinoline (8TQ, IC 50 value ∼2.5 μM) displayed strong inhibition of Rpn11. Further synthetic elaboration of 8TQ yielded a small molecule compound (35, IC 50 value ∼400 nM) that is a potent and selective inhibitor of Rpn11 that blocks proliferation of tumor cells in culture.

  20. Bioactive compounds of fresh and dehydrated green pepper

    Directory of Open Access Journals (Sweden)

    Ana Marinho do Nascimento

    2017-07-01

    Full Text Available Pepper Capsicum annuum L., belongs to the Solanaceae family, which contains approximately 31 species. Bioactive compounds also known as phytochemicals are chemical and biochemical components that are present in most fruits and vegetables. The objective of the present study was to verify if the bioactive compounds of the green pepper remain after being submitted to the drying process. The experiment was conducted in a completely randomized design with 2 treatments and 5 replicates. Green peppers were used from the (Economic Center of Supply Corporation of the city of Patos, Paraíba. The peppers were packed in plastic boxes and transported to the Laboratory of Chemistry, Biochemistry and Food Analysis of the Federal University of Campina Grande, Campus Pombal. Where they were selected, washed and sanitized. After that, the minimum processing was done and the drying was carried out in a circulation oven at 60 ºC. At the end of the drying, the samples were crushed and sieved. After this process, the analyzes of ascorbic acid, chlorophylls, carotenoids, anthocyanin flavonoids and phenolic compounds. It was found that there was a significant difference between treatments. The bioactive properties of green pepper were not lost after the heat treatment. Some phytochemicals as ascorbic acid, carotenoids and phenolic compounds were concentrated. Therefore the loss of water during the drying process increased the concentration of the bioactive compounds of dehydrated pepper, the product obtained with this method exhibited high levels of phytochemicals, the use of drying may be an alternative to prolong the shelf life of the vegetable.

  1. Involvement of Proteasome and Macrophages M2 in the Protection Afforded by Telmisartan against the Acute Myocardial Infarction in Zucker Diabetic Fatty Rats with Metabolic Syndrome

    Directory of Open Access Journals (Sweden)

    C. Di Filippo

    2014-01-01

    Full Text Available This study investigated the involvement of proteasome and macrophages M2 in the protection afforded by telmisartan against the acute myocardial infarction in Zucker diabetic fatty (ZDF rats with metabolic syndrome. ZDF rats were treated for three weeks with telmisartan at doses of 7 and 12 mg/kg/day. After treatment, rats were subjected to a 25 min occlusion of the left descending coronary artery followed by 2 h reperfusion (I/R. At the end of the I/R period, biochemical, immunohistochemical, and echocardiographic evaluations were done. Telmisartan treatment (7 mg/kg and 12 mg/kg reduced the myocardial infarct size, the expression of proteasome subunits 20S and 26S, and the protein ubiquitin within the heart. The compound has led to an increased M2 macrophage phenotype within the cardiac specimens and a modification of the cardiac cytokine and chemokine profile. This was functionally translated in improved cardiac performance as evidenced by echography after 2 h reperfusion. 7 mg/kg/day telmisartan was sufficient to improve the left ventricular ejection fraction LVEF of the rat heart recorded after I/R (e.g., vehicle 38 ± 2.2%; telmisartan 54 ± 2.7% and was sufficient to improve the diastolic function and the myocardial performance index up to values of 0.6 ± 0.01 measured after I/R.

  2. Ubiquitin-proteasomal degradation of COX-2 in TGF-β stimulated human endometrial cells is mediated through endoplasmic reticulum mannosidase I.

    Science.gov (United States)

    Singh, Mohan; Chaudhry, Parvesh; Parent, Sophie; Asselin, Eric

    2012-01-01

    Cyclooxygenase (COX)-2 is a key regulatory enzyme in the production of prostaglandins (PG) during various physiological processes. Mechanisms of COX-2 regulation in human endometrial stromal cells (human endometrial stromal cells) are not fully understood. In this study, we investigate the role of TGF-β in the regulation of COX-2 in human uterine stromal cells. Each TGF-β isoform decreases COX-2 protein level in human uterine stromal cells in Smad2/3-dependent manner. The decrease in COX-2 is accompanied by a decrease in PG synthesis. Knockdown of Smad4 using specific small interfering RNA prevents the decrease in COX-2 protein, confirming that Smad pathway is implicated in the regulation of COX-2 expression in human endometrial stromal cells. Pretreatment with 26S proteasome inhibitor, MG132, significantly restores COX-2 protein and PG synthesis, indicating that COX-2 undergoes proteasomal degradation in the presence of TGF-β. In addition, each TGF-β isoform up-regulates endoplasmic reticulum (ER)-mannosidase I (ERManI) implying that COX-2 degradation is mediated through ER-associated degradation pathway in these cells. Furthermore, inhibition of ERManI activity using the mannosidase inhibitor (kifunensine), or small interfering RNA-mediated knockdown of ERManI, prevents TGF-β-induced COX-2 degradation. Taken together, these studies suggest that TGF-β promotes COX-2 degradation in a Smad-dependent manner by up-regulating the expression of ERManI and thereby enhancing ER-associated degradation and proteasomal degradation pathways.

  3. Applicability of viscosity measurement to the detection of irradiated peppers

    International Nuclear Information System (INIS)

    Hayashi, T.; Todoriki, S.; Kohyama, K.

    1996-01-01

    Starch is degraded by ionising radiation, resulting in a decrease in viscosity. The viscosities of black and white peppers which contain large amounts of starch are reduced by irradiation so, therefore, viscosity measurement has been proposed as a method to detect the irradiation treatment of these food products. Although detection of irradiated spices by thermoluminescence measurement has been established, it is useful to establish the viscosity measuring technique for detecting irradiated peppers, as this method is carried out widely in the laboratories of food controlling authorities and food processing companies. (author)

  4. Washing effects of limonene on pesticide residues in green peppers.

    Science.gov (United States)

    Lu, Hai-Yan; Shen, Yan; Sun, Xing; Zhu, Hong; Liu, Xian-Jin

    2013-09-01

    The presence of pesticide residues in food has caused much concern. The low health risks and environmental impacts of limonene make it a very interesting solvent for use in green chemistry. Washing effects of limonene on pesticide residues of methyl chlorpyrifos, chlorothalonil, chlorpyrifos, fenpropathrin and deltamethrin were investigated in green pepper. Results showed that washing with a low concentration of limonene for 5 min (where LOQ is limit of quantitation) caused 53.67%, limonene for 10 min produced 55.90%, limonene for 5 min was the optimal treatment for elimination of pesticide residues in green pepper, considering effect and treatment time as well as cost. © 2013 Society of Chemical Industry.

  5. Virus diseases of peppers (Capsicum spp.) and their control.

    Science.gov (United States)

    Kenyon, Lawrence; Kumar, Sanjeet; Tsai, Wen-Shi; Hughes, Jacqueline d'A

    2014-01-01

    The number of virus species infecting pepper (Capsicum spp.) crops and their incidences has increased considerably over the past 30 years, particularly in tropical and subtropical pepper production systems. This is probably due to a combination of factors, including the expansion and intensification of pepper cultivation in these regions, the increased volume and speed of global trade of fresh produce (including peppers) carrying viruses and vectors to new locations, and perhaps climate change expanding the geographic range suitable for the viruses and vectors. With the increased incidences of diverse virus species comes increased incidences of coinfection with two or more virus species in the same plant. There is then greater chance of synergistic interactions between virus species, increasing symptom severity and weakening host resistance, as well as the opportunity for genetic recombination and component exchange and a possible increase in aggressiveness, virulence, and transmissibility. The main virus groups infecting peppers are transmitted by aphids, whiteflies, or thrips, and a feature of many populations of these vector groups is that they can develop resistance to some of the commonly used insecticides relatively quickly. This, coupled with the increasing concern over the impact of over- or misuse of insecticides on the environment, growers, and consumers, means that there should be less reliance on insecticides to control the vectors of viruses infecting pepper crops. To improve the durability of pepper crop protection measures, there should be a shift away from the broadscale use of insecticides and the use of single, major gene resistance to viruses. Instead, integrated and pragmatic virus control measures should be sought that combine (1) cultural practices that reduce sources of virus inoculum and decrease the rate of spread of viruliferous vectors into the pepper crop, (2) synthetic insecticides, which should be used judiciously and only when the

  6. Chemical constituents of pungent spice pepper (Capsicum annuum L.) from Macedonian origin

    OpenAIRE

    Rafajlovska, Vesna; Slaveska-Raicki, Renata; Koleva Gudeva, Liljana; Mitrev, Sasa; Srbinoska, Marija

    2004-01-01

    In this paper the chemical constituents of the pungent spice pepper Capsicum annuum L.ssp. Microcarpum from Macedonian origin are estimated. Content of moisture, proteins and soluble sugar is 9.60% and 20.33%, respectively. Color capacity of the pungent spice pepper is 5.60g capsanthin/kg pepper dry matter. The influence of organic solvents on the pepper oleoresin extraction and contents of colored components and capsaicin content in it is also studied. The highest quantity of pepper oleor...

  7. Proteasomal control of cytokinin synthesis protects Mycobacterium tuberculosis against nitric oxide

    Science.gov (United States)

    Samanovic, Marie I.; Tu, Shengjiang; Novák, Ondřej; Iyer, Lakshminarayan M.; McAllister, Fiona E.; Aravind, L.; Gygi, Steven P.; Hubbard, Stevan R.; Strnad, Miroslav; Darwin, K. Heran

    2015-01-01

    Summary One of several roles of the Mycobacterium tuberculosis proteasome is to defend against host-produced nitric oxide (NO), a free radical that can damage numerous biological macromolecules. Mutations that inactivate proteasomal degradation in Mycobacterium tuberculosis result in bacteria that are hypersensitive to NO and attenuated for growth in vivo, but it was not known why. To elucidate the link between proteasome function, NO-resistance, and pathogenesis, we screened for suppressors of NO hypersensitivity in a mycobacterial proteasome ATPase mutant and identified mutations in Rv1205. We determined that Rv1205 encodes a pupylated proteasome substrate. Rv1205 is a homologue of the plant enzyme LONELY GUY, which catalyzes the production of hormones called cytokinins. Remarkably, we report for the first time that an obligate human pathogen secretes several cytokinins. Finally, we determined that the Rv1205-dependent accumulation of cytokinin breakdown products is likely responsible for the sensitization of Mycobacterium tuberculosis proteasome-associated mutants to NO. PMID:25728768

  8. The Ubiquitin-associated (UBA) 1 Domain of Schizosaccharomyces pombe Rhp23 Is Essential for the Recognition of Ubiquitin-proteasome System Substrates Both in Vitro and in Vivo*

    Science.gov (United States)

    Medina, Bethan; Paraskevopoulos, Konstantinos; Boehringer, Jonas; Sznajder, Anna; Robertson, Morag; Endicott, Jane; Gordon, Colin

    2012-01-01

    The ubiquitin-proteasome system is essential for maintaining a functional cell. Not only does it remove incorrectly folded proteins, it also regulates protein levels to ensure their appropriate spatial and temporal distribution. Proteins marked for degradation by the addition of Lys48-linked ubiquitin (Ub) chains are recognized by shuttle factors and transported to the 26 S proteasome. One of these shuttle factors, Schizosaccharomyces pombe Rhp23, has an unusual domain architecture. It comprises an N-terminal ubiquitin-like domain that can recognize the proteasome followed by two ubiquitin-associated (UBA) domains, termed UBA1 and UBA2, which can bind Ub. This architecture is conserved up to humans, suggesting that both domains are important for Rhp23 function. Such an extent of conservation raises the question as to why, in contrast to all other shuttle proteins, does Rhp23 require two UBA domains? We performed in vitro Ub binding assays using domain swap chimeric proteins and mutated domains in isolation as well as in the context of the full-length protein to reveal that the Ub binding properties of the UBA domains are context-dependent. In vivo, the internal Rhp23 UBA1 domain provides sufficient Ub recognition for the protein to function without UBA2. PMID:23038266

  9. Changes in expression of proteasome in rats at different stages of atherosclerosis.

    Science.gov (United States)

    Ismawati; Oenzil, Fadil; Yanwirasti; Yerizel, Eti

    2016-06-01

    It has been suggested that proteasome system has a role in initiation, progression, and complication stages of atherosclerosis. Although there is still controversy, there has been no research that compares the expression of proteasome in tissue and serum at each of these stages. This study aimed to investigated the expression of proteasome at different stages of atherosclerosis using rat model. We measured the expression of aortic proteasome by immunohistochemical analyses and were then analyzed using ImageJ software for percentage of area and integrated density. We used Photoshop version 3.0 to analyze aortic proteasome expression as a comparison. We measured serum proteasome expression by enzyme linked immunosorbents assays. Kruskal-Wallis test was used to compare mean value of percentage of area and serum proteasome. Analysis of variance test was used to compare mean value of integrated density. Correlation test between vascular proteasome expression and serum proteasome expression was made using Spearman test. A P-value of 0.05 was considered statistically significant. Compared with normal, percentage of area was higher in initiation, progression, and complication. Compared with normal, integrated density was higher in initiation and further higher in progression and complication. Data from Image J is similar with data from Photoshop. Serum proteasome expression was higher in initiation compared with normal, and further higher in progression and complication. It was concluded that there were different vascular proteasome expression and serum proteasome expression at the stages of atherosclerosis. These results may be used in research into new marker and therapeutic target in atherosclerosis.

  10. Mona F1: New pepper (Capsicum annuum L. hybrid in the Centre for Vegetable Crops

    Directory of Open Access Journals (Sweden)

    Cvikić Dejan

    2007-01-01

    Full Text Available The planted area various ways of pepper consumption (fresh or processed, make pepper one of the most important cultivars in vegetable breeding. In our country, up until now, the producers have usually grown varieties and domestic populations of pepper, while in more developed countries the usage of F1 hybrids is much more popular. The first pepper hybrids have been created in the Centre for Vegetable Crops by crossing new lines with male sterility gene ms-3 and selected genotypes from pepper collection. Created hybrids have higher yield, quality fruits and early ripening. This paper is the result of comparative trial in controlled conditions. Pepper varieties Župska rana, Zlatna medalja, Palanačka kapija and Duga bela, as well as new hybrid Mona F1 were the research matherial in order to observe the most important pepper traits.

  11. Physical and Sensory Properties of Ice Cream Containing Fermented Pepper Powder.

    Science.gov (United States)

    Yeon, Su-Jung; Kim, Ji-Han; Hong, Go-Eun; Park, Woojoon; Kim, Soo-Ki; Seo, Han-Geuk; Lee, Chi-Ho

    2017-01-01

    This study was conducted to investigate the physical and sensory properties of ice cream containing fermented pepper powder. Three ice cream formulas were manufactured: 1, control; 2, supplemented with 0.1% fermented pepper powder; and 3, supplemented with 0.2% fermented pepper powder. Formulas 2 and 3 had significantly higher viscosity and lower overrun than formula 1 ( p ice creams supplemented with fermented pepper powder were harder and maintained their forms longer than the controls. 0.2% fermented pepper powder added ice cream had no pungency as much as that of control and overall sensory attribute was not significantly different from control. Therefore, ice cream containing fermented pepper powder maintained physical and sensory properties similar to the controls, and maintenance was better. It means fermented pepper powder ice cream can be utilized as the material of functional food (dessert).

  12. streamgap-pepper: Effects of peppering streams with many small impacts

    Science.gov (United States)

    Bovy, Jo; Erkal, Denis; Sanders, Jason

    2017-02-01

    streamgap-pepper computes the effect of subhalo fly-bys on cold tidal streams based on the action-angle representation of streams. A line-of-parallel-angle approach is used to calculate the perturbed distribution function of a given stream segment by undoing the effect of all impacts. This approach allows one to compute the perturbed stream density and track in any coordinate system in minutes for realizations of the subhalo distribution down to 10^5 Msun, accounting for the stream's internal dispersion and overlapping impacts. This code uses galpy (ascl:1411.008) and the streampepperdf.py galpy extension, which implements the fast calculation of the perturbed stream structure.

  13. Comparative analysis of pepper and tomato reveals euchromatin expansion of pepper genome caused by differential accumulation of Ty3/Gypsy-like elements

    Directory of Open Access Journals (Sweden)

    Ahn Jong Hwa

    2011-01-01

    Full Text Available Abstract Background Among the Solanaceae plants, the pepper genome is three times larger than that of tomato. Although the gene repertoire and gene order of both species are well conserved, the cause of the genome-size difference is not known. To determine the causes for the expansion of pepper euchromatic regions, we compared the pepper genome to that of tomato. Results For sequence-level analysis, we generated 35.6 Mb of pepper genomic sequences from euchromatin enriched 1,245 pepper BAC clones. The comparative analysis of orthologous gene-rich regions between both species revealed insertion of transposons exclusively in the pepper sequences, maintaining the gene order and content. The most common type of the transposon found was the LTR retrotransposon. Phylogenetic comparison of the LTR retrotransposons revealed that two groups of Ty3/Gypsy-like elements (Tat and Athila were overly accumulated in the pepper genome. The FISH analysis of the pepper Tat elements showed a random distribution in heterochromatic and euchromatic regions, whereas the tomato Tat elements showed heterochromatin-preferential accumulation. Conclusions Compared to tomato pepper euchromatin doubled its size by differential accumulation of a specific group of Ty3/Gypsy-like elements. Our results could provide an insight on the mechanism of genome evolution in the Solanaceae family.

  14. Focal dysfunction of the proteasome: a pathogenic factor in a mouse model of amyotrophic lateral sclerosis.

    Science.gov (United States)

    Kabashi, Edor; Agar, Jeffrey N; Taylor, David M; Minotti, Sandra; Durham, Heather D

    2004-06-01

    Mutations in the Cu/Zn-superoxide dismutase (SOD-1) gene are responsible for a familial form of amyotrophic lateral sclerosis (fALS). The present study demonstrated impaired proteasomal function in the lumbar spinal cord of transgenic mice expressing human SOD-1 with the ALS-causing mutation G93A (SOD-1(G93A)) compared to non-transgenic littermates (LM) and SOD-1(WT) transgenic mice. Chymotrypsin-like activity was decreased as early as 45 days of age. By 75 days, chymotrypsin-, trypsin-, and caspase-like activities of the proteasome were impaired, at about 50% of control activity in lumbar spinal cord, but unchanged in thoracic spinal cord and liver. Both total and specific activities of the proteasome were reduced to a similar extent, indicating that a change in proteasome function, rather than a decrease in proteasome levels, had occurred. Similar decreases of total and specific activities of the proteasome were observed in NIH 3T3 cell lines expressing fALS mutants SOD-1(G93A) and SOD-1(G41S), but not in SOD-1(WT) controls. Although overall levels of proteasome were maintained in spinal cord of SOD-1(G93A) transgenic mice, the level of 20S proteasome was substantially reduced in lumbar spinal motor neurons relative to the surrounding neuropil. It is concluded that impairment of the proteasome is an early event and contributes to ALS pathogenesis.

  15. Inhibitors of the proteasome suppress homologous DNA recombination in mammalian cells.

    Science.gov (United States)

    Murakawa, Yasuhiro; Sonoda, Eiichiro; Barber, Louise J; Zeng, Weihua; Yokomori, Kyoko; Kimura, Hiroshi; Niimi, Atsuko; Lehmann, Alan; Zhao, Guang Yu; Hochegger, Helfrid; Boulton, Simon J; Takeda, Shunichi

    2007-09-15

    Proteasome inhibitors are novel antitumor agents against multiple myeloma and other malignancies. Despite the increasing clinical application, the molecular basis of their antitumor effect has been poorly understood due to the involvement of the ubiquitin-proteasome pathway in multiple cellular metabolisms. Here, we show that treatment of cells with proteasome inhibitors has no significant effect on nonhomologous end joining but suppresses homologous recombination (HR), which plays a key role in DNA double-strand break (DSB) repair. In this study, we treat human cells with proteasome inhibitors and show that the inhibition of the proteasome reduces the efficiency of HR-dependent repair of an artificial HR substrate. We further show that inhibition of the proteasome interferes with the activation of Rad51, a key factor for HR, although it does not affect the activation of ATM, gammaH2AX, or Mre11. These data show that the proteasome-mediated destruction is required for the promotion of HR at an early step. We suggest that the defect in HR-mediated DNA repair caused by proteasome inhibitors contributes to antitumor effect, as HR plays an essential role in cellular proliferation. Moreover, because HR plays key roles in the repair of DSBs caused by chemotherapeutic agents such as cisplatin and by radiotherapy, proteasome inhibitors may enhance the efficacy of these treatments through the suppression of HR-mediated DNA repair pathways.

  16. Proteasome function is not impaired in healthy aging of the lung.

    Science.gov (United States)

    Caniard, Anne; Ballweg, Korbinian; Lukas, Christina; Yildirim, Ali Ö; Eickelberg, Oliver; Meiners, Silke

    2015-10-01

    Aging is the progressive loss of cellular function which inevitably leads to death. Failure of proteostasis including the decrease in proteasome function is one hallmark of aging. In the lung, proteasome activity was shown to be impaired in age-related diseases such as chronic obstructive pulmonary disease. However, little is known on proteasome function during healthy aging. Here, we comprehensively analyzed healthy lung aging and proteasome function in wildtype, proteasome reporter and immunoproteasome knockout mice. Wildtype mice spontaneously developed senile lung emphysema while expression and activity of proteasome complexes and turnover of ubiquitinated substrates was not grossly altered in lungs of aged mice. Immunoproteasome subunits were specifically upregulated in the aged lung and the caspase-like proteasome activity concomitantly decreased. Aged knockout mice for the LMP2 or LMP7 immunoproteasome subunits showed no alteration in proteasome activities but exhibited typical lung aging phenotypes suggesting that immunoproteasome function is dispensable for physiological lung aging in mice. Our results indicate that healthy aging of the lung does not involve impairment of proteasome function. Apparently, the reserve capacity of the proteostasis systems in the lung is sufficient to avoid severe proteostasis imbalance during healthy aging.

  17. Modulation of Protein Quality Control and Proteasome to Autophagy Switch in Immortalized Myoblasts from Duchenne Muscular Dystrophy Patients

    Directory of Open Access Journals (Sweden)

    Marion Wattin

    2018-01-01

    Full Text Available The maintenance of proteome integrity is of primary importance in post-mitotic tissues such as muscle cells; thus, protein quality control mechanisms must be carefully regulated to ensure their optimal efficiency, a failure of these processes being associated with various muscular disorders. Duchenne muscular dystrophy (DMD is one of the most common and severe forms of muscular dystrophies and is caused by mutations in the dystrophin gene. Protein quality control modulations have been diversely observed in degenerating muscles of patients suffering from DMD or in animal models of the disease. In this study, we investigated whether modulations of protein quality control mechanisms already pre-exist in undifferentiated myoblasts originating from DMD patients. We report for the first time that the absence of dystrophin in human myoblasts is associated with protein aggregation stress characterized by an increase of protein aggregates. This stress is combined with BAG1 to BAG3 switch, NFκB activation and up-regulation of BAG3/HSPB8 complexes that ensure preferential routing of misfolded/aggregated proteins to autophagy rather than to deficient 26S proteasome. In this context, restoration of pre-existing alterations of protein quality control processes might represent an alternative strategy for DMD therapies.

  18. [Evaluation of the mercury accumulating capacity of pepper (Capsicum annuum)].

    Science.gov (United States)

    Pérez-Vargas, Híver M; Vidal-Durango, Jhon V; Marrugo-Negrete, José L

    2014-01-01

    To assess the mercury accumulating capacity in contaminated soils from the community of Mina Santa Cruz, in the south of the department of Bolívar, Colombia, of the pepper plant (Capsicum annuum), in order to establish the risk to the health of the consuming population. Samples were taken from tissues (roots, stems, and leaves) of pepper plants grown in two soils contaminated with mercury and a control soil during the first five months of growth to determine total mercury through cold vapor atomic absorption spectrometry. Total mercury was determined in the samples of pepper plant fruits consumed in Mina Santa Cruz. The mean concentrations of total mercury in the roots were higher than in stems and leaves. Accumulation in tissues was influenced by mercury levels in soil and the growth time of the plants. Mercury concentrations in fruits of pepper plant were lower than tolerable weekly intake provided by WHO. Percent of translocation of mercury to aerial parts of the plant were low in both control and contaminated soils. Despite low levels of mercury in this food, it is necessary to minimize the consumption of food contaminated with this metal.

  19. Growth and physiological aspects of bell pepper ( Capsicum annuum )

    African Journals Online (AJOL)

    This study aimed to evaluate growth and physiological aspects of 'All Big' bell pepper, under saline stress and exogenous application of proline on the leaves. The research was conducted in pots adapted as drainage lysimeters under greenhouse conditions, using sandy-loam eutrophic Regolithic Neosol, in the ...

  20. Influence of turmeric rhizome and black pepper on blood ...

    African Journals Online (AJOL)

    ) and black pepper (BP) on blood components and performance of male broiler chickens. A 2 × 3 factorial arrangement with two levels of TRP (0 and 0.5 g/kg) and three levels of BP (0, 0.5 and 1 g/kg) were used to provide six dietary treatments ...

  1. High-throughput sequencing of black pepper root transcriptome

    Science.gov (United States)

    2012-01-01

    Background Black pepper (Piper nigrum L.) is one of the most popular spices in the world. It is used in cooking and the preservation of food and even has medicinal properties. Losses in production from disease are a major limitation in the culture of this crop. The major diseases are root rot and foot rot, which are results of root infection by Fusarium solani and Phytophtora capsici, respectively. Understanding the molecular interaction between the pathogens and the host’s root region is important for obtaining resistant cultivars by biotechnological breeding. Genetic and molecular data for this species, though, are limited. In this paper, RNA-Seq technology has been employed, for the first time, to describe the root transcriptome of black pepper. Results The root transcriptome of black pepper was sequenced by the NGS SOLiD platform and assembled using the multiple-k method. Blast2Go and orthoMCL methods were used to annotate 10338 unigenes. The 4472 predicted proteins showed about 52% homology with the Arabidopsis proteome. Two root proteomes identified 615 proteins, which seem to define the plant’s root pattern. Simple-sequence repeats were identified that may be useful in studies of genetic diversity and may have applications in biotechnology and ecology. Conclusions This dataset of 10338 unigenes is crucially important for the biotechnological breeding of black pepper and the ecogenomics of the Magnoliids, a major group of basal angiosperms. PMID:22984782

  2. Poultry manure application and varietal effects of chilli-pepper ...

    African Journals Online (AJOL)

    Cultural practices such as organic manure application can affect soil fertility and also insect pest and disease incidence on the plant. The effect of poultry manure application was therefore evaluated in relation to the infestation by major insect pests and disease of pepper in a humid tropical agro-ecosystem. Treatments ...

  3. Recent approaches for bell pepper seed vigor testing

    Directory of Open Access Journals (Sweden)

    Marcos Altomani Neves Dias

    2014-08-01

    Full Text Available Seed vigor is a major attribute in the determination of whether a seed lot can be commercialized. Different physiological tests are used to assess seed vigor and are generally based on different concepts, which could lead to variable interpretations, particularly when comparing lots presenting different vigor levels. This study was conducted to compare the different approaches used to evaluate bell pepper seed vigor and verify the applicability of the Seed Vigor Imaging System (SVIS for this purpose. The experiment was performed using four seed lots of bell pepper hybrids in two periods, and the evaluations involved germination, seedling emergence, seedling emergence speed index, saturated salt accelerated aging and image analysis. The use of seedling imaging analysis by SVISis a new approach for bell pepper seed vigor evaluation. Both the software and traditional tests provided sufficient information for seed vigor analysis, enabling the segregation of the analyzed seed lots. Thus, the new method fulfills the requirement for an efficient, rapid, cost-effective and standardized method that allows comparisons among laboratories and avoids human influence on the interpretation of the results. SVIS can be considered a feasible option to be included in quality assurance programs of the bell pepper seed industry.

  4. High-throughput sequencing of black pepper root transcriptome

    Directory of Open Access Journals (Sweden)

    Gordo Sheila MC

    2012-09-01

    Full Text Available Abstract Background Black pepper (Piper nigrum L. is one of the most popular spices in the world. It is used in cooking and the preservation of food and even has medicinal properties. Losses in production from disease are a major limitation in the culture of this crop. The major diseases are root rot and foot rot, which are results of root infection by Fusarium solani and Phytophtora capsici, respectively. Understanding the molecular interaction between the pathogens and the host’s root region is important for obtaining resistant cultivars by biotechnological breeding. Genetic and molecular data for this species, though, are limited. In this paper, RNA-Seq technology has been employed, for the first time, to describe the root transcriptome of black pepper. Results The root transcriptome of black pepper was sequenced by the NGS SOLiD platform and assembled using the multiple-k method. Blast2Go and orthoMCL methods were used to annotate 10338 unigenes. The 4472 predicted proteins showed about 52% homology with the Arabidopsis proteome. Two root proteomes identified 615 proteins, which seem to define the plant’s root pattern. Simple-sequence repeats were identified that may be useful in studies of genetic diversity and may have applications in biotechnology and ecology. Conclusions This dataset of 10338 unigenes is crucially important for the biotechnological breeding of black pepper and the ecogenomics of the Magnoliids, a major group of basal angiosperms.

  5. Molecular characterization and diversity analysis in chilli pepper ...

    African Journals Online (AJOL)

    India is considered to be the secondary center of diversity of chilli pepper, especially of Capsicum annuum. Simple sequence repeats (SSRs) are the most widely used marker system for plant variety characterization and diversity analysis especially in cultivated species which have low levels of polymorphism. The diversity ...

  6. Influence of turmeric rhizome and black pepper on blood ...

    African Journals Online (AJOL)

    2012-04-26

    ) and black pepper. (BP) on blood components and performance of male broiler chickens. A 2 × 3 factorial arrangement with two levels of TRP (0 and 0.5 g/kg) and three levels of BP (0, 0.5 and 1 g/kg) were used to provide six.

  7. Detection and Quantification of Leveillula taurica Growth in Pepper Leaves

    NARCIS (Netherlands)

    Zheng, Z.; Nonomura, T.; Bóka, K.; Matsuda, Y.; Visser, R.G.F.; Toyoda, H.; Kiss, L.; Bai, Y.

    2013-01-01

    Leveillula taurica is an obligate fungal pathogen that causes powdery mildew disease on a broad range of plants, including important crops such as pepper, tomato, eggplant, onion, cotton, and so on. The early stage of this disease is difficult to diagnose and the disease can easily spread

  8. Breeding for pepper fruit quality: a genitical metabolomics approach

    NARCIS (Netherlands)

    Wahyuni, Y.

    2014-01-01

    A diverse collection of 32 pepper accessions was analysed for variation in health-related metabolites, such as carotenoids, capsaicinoids, flavonoids and vitamins C and E. For each of the metabolites analysed, there was a lot of variation among the accessions and it was possible to

  9. Variation in Yugoslavian hot pepper (Capsicum annuum L.) accessions.

    NARCIS (Netherlands)

    Zeven, A.C.; Zewdie, Y.

    1997-01-01

    A total of 67 hot pepper accessions were evaluated based on 35 morphological and physiological characters. Differences were observed in a number of characters. The accessions were grouped into six clusters, mainly based on fruit weight, 1000 seed weight, and fruit number per plant. accessions -

  10. Comparative studies of ginger ( Zingiber officinale ) and black pepper

    African Journals Online (AJOL)

    Extracts of two spices namely ginger (Zingiber officinale) and black pepper (Piper guinenses) were prepared in 0.4, 1.2, 2.4 and 3.6% concentrations. Soymilk and kunuzaki, were treated, respectively with the different concentrations and stored at ambient temperature for 5 days. The microbial load and identification were ...

  11. Effect of aqueous extract of alligator pepper (Zingiberaceae ...

    African Journals Online (AJOL)

    Twenty female and ten male Sprague-Dawley rats of proven fertility from a pilot study were randomly mated in groups of two females and one male. Three days later, female rats in the ... weight gain reduction. Key words: Gestational weight gain, Aqueous Extract, Intra-peritoneal injection, Alligator Pepper, Nutrition.

  12. Fulvic acid affects pepper antioxidant activity and fruit quality ...

    African Journals Online (AJOL)

    Fulvic acid has been considered as a valuable fertilizer for sustainable agriculture. The present investigation was undertaken to evaluate the effect of fulvic acid (FA) on antioxidant compounds and fruit quality of pepper under field conditions. Plants were grown in the Department of Horticulture Farm, Ferdowsi University of ...

  13. Structure and function based design of Plasmodium-selective proteasome inhibitors

    Science.gov (United States)

    Li, Hao; O'Donoghue, Anthony J.; van der Linden, Wouter A.; Xie, Stanley C.; Yoo, Euna; Foe, Ian T.; Tilley, Leann; Craik, Charles S.; da Fonseca, Paula C. A.; Bogyo, Matthew

    2016-01-01

    The proteasome is a multi-component protease complex responsible for regulating key processes such as the cell cycle and antigen presentation1. Compounds that target the proteasome are potentially valuable tools for the treatment of pathogens that depend on proteasome function for survival and replication. In particular, proteasome inhibitors have been shown to be toxic for the malaria parasite Plasmodium falciparum at all stages of its life cycle2-5. Most compounds that have been tested against the parasite also inhibit the mammalian proteasome resulting in toxicity that precludes their use as therapeutic agents2,6. Therefore, better definition of the substrate specificity and structural properties of the Plasmodium proteasome could enable the development of compounds with sufficient selectivity to allow their use as anti-malarial agents. To accomplish this goal, we used a substrate profiling method to uncover differences in the specificities of the human and P. falciparum proteasome. We designed inhibitors based on amino acid preferences specific to the parasite proteasome, and found that they preferentially inhibit the β 2 subunit. We determined the structure of the P. falciparum 20S proteasome bound to the inhibitor using cryo-electron microscopy (cryo-EM) and single particle analysis, to a resolution of 3.6 Å. These data reveal the unusually open P. falciparum β2 active site and provide valuable information regarding active site architecture that can be used to further refine inhibitor design. Furthermore, consistent with the recent finding that the proteasome is important for stress pathways associated with resistance of artemisinin (ART) family anti-malarials7,8, we observed growth inhibition synergism with low doses of this β 2 selective inhibitor in ART sensitive and resistant parasites. Finally, we demonstrated that a parasite selective inhibitor could be used to attenuate parasite growth in vivo without significant toxicity to the host. Thus, the

  14. Structure- and function-based design of Plasmodium-selective proteasome inhibitors.

    Science.gov (United States)

    Li, Hao; O'Donoghue, Anthony J; van der Linden, Wouter A; Xie, Stanley C; Yoo, Euna; Foe, Ian T; Tilley, Leann; Craik, Charles S; da Fonseca, Paula C A; Bogyo, Matthew

    2016-02-11

    The proteasome is a multi-component protease complex responsible for regulating key processes such as the cell cycle and antigen presentation. Compounds that target the proteasome are potentially valuable tools for the treatment of pathogens that depend on proteasome function for survival and replication. In particular, proteasome inhibitors have been shown to be toxic for the malaria parasite Plasmodium falciparum at all stages of its life cycle. Most compounds that have been tested against the parasite also inhibit the mammalian proteasome, resulting in toxicity that precludes their use as therapeutic agents. Therefore, better definition of the substrate specificity and structural properties of the Plasmodium proteasome could enable the development of compounds with sufficient selectivity to allow their use as anti-malarial agents. To accomplish this goal, here we use a substrate profiling method to uncover differences in the specificities of the human and P. falciparum proteasome. We design inhibitors based on amino-acid preferences specific to the parasite proteasome, and find that they preferentially inhibit the β2-subunit. We determine the structure of the P. falciparum 20S proteasome bound to the inhibitor using cryo-electron microscopy and single-particle analysis, to a resolution of 3.6 Å. These data reveal the unusually open P. falciparum β2 active site and provide valuable information about active-site architecture that can be used to further refine inhibitor design. Furthermore, consistent with the recent finding that the proteasome is important for stress pathways associated with resistance of artemisinin family anti-malarials, we observe growth inhibition synergism with low doses of this β2-selective inhibitor in artemisinin-sensitive and -resistant parasites. Finally, we demonstrate that a parasite-selective inhibitor could be used to attenuate parasite growth in vivo without appreciable toxicity to the host. Thus, the Plasmodium proteasome is a

  15. Targeting neuroblastoma stem cells with retinoic acid and proteasome inhibitor.

    Directory of Open Access Journals (Sweden)

    Barbara Hämmerle

    Full Text Available Neuroblastma cell lines contain a side-population of cells which express stemness markers. These stem-like cells may represent the potential underlying mechanism for resistance to conventional therapy and recurrence of neuroblastoma in patients.To develop novel strategies for targeting the side-population of neurobastomas, we analyzed the effects of 13-cis-retinoic acid (RA combined with the proteasome inhibitor MG132. The short-term action of the treatment was compared with effects after a 5-day recovery period during which both chemicals were withdrawn. RA induced growth arrest and differentiation of SH-SY5Y and SK-N-BE(2 neuroblastoma cell lines. Inhibition of the proteasome caused apoptosis in both cell lines, thus, revealing the critical role of this pathway in the regulated degradation of proteins involved in neuroblastoma proliferation and survival. The combination of RA with MG132 induced apoptosis in a dose-dependent manner, in addition to promoting G2/M arrest in treated cultures. Interestingly, expression of stem cell markers such as Nestin, Sox2, and Oct4 were reduced after the recovery period of combined treatment as compared with untreated cells or treated cells with either compound alone. Consistent with this, neurosphere formation was significantly impaired by the combined treatment of RA and MG132.Given that stem-like cells are associated with resistant to conventional therapy and are thought to be responsible for relapse, our results suggest that dual therapy of RA and proteasome inhibitor might be beneficial for targeting the side-population of cells associated residual disease in high-risk neuroblastoma.

  16. Effects of Radiation on Proteasome Function in Prostate Cancer Cells

    Science.gov (United States)

    2009-02-01

    precursor [ Homo sapiens ] BCL2-associated athanogene 3 adhesion regulating molecule 1 precursor CD44 antigen heat shock 27kDa protein 1 DnaJ... feature of CICs that can easily be exploited to identify , track, and target them in vitro and in vivo . J Natl Cancer Inst 2009;101:1–10 Q1 Q5 JNCI...P = .055; Supplementary Figure 1, D and E, available online), indicating that reduced proteasome activity in CICs is a feature found across tumor

  17. Potential roles for ubiquitin and the proteasome during ribosome biogenesis

    Czech Academy of Sciences Publication Activity Database

    Stavreva, D. A.; Kawasaki, M.; Dundr, M.; Koberna, Karel; Müller, W. G.; Tsujimura-Takahashi, T.; Komatsu, W.; Hayano, T.; Isobe, T.; Raška, Ivan; Misteli, T.; Takahashi, N.; McNally, J. G.

    2006-01-01

    Roč. 26, č. 13 (2006), s. 5131-5145 ISSN 0270-7306 R&D Projects: GA MŠk(CZ) LC535; GA ČR(CZ) GA304/05/0374; GA ČR(CZ) GA304/04/0692 Grant - others:NIH(US) Intramural Research Program; Ministry of Education(JP) Pioneer Research grant Institutional research plan: CEZ:AV0Z50110509 Keywords : the role of the ubikvitin * proteasome system in ribosome biogenesis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.773, year: 2006

  18. Imaging Reporters for Proteasome Activity Identify Tumor- and Metastasis-Initiating Cells

    Directory of Open Access Journals (Sweden)

    Amanda C. Stacer

    2015-08-01

    Full Text Available Tumor-initiating cells, also designated as cancer stem cells, are proposed to constitute a subpopulation of malignant cells central to tumorigenesis, metastasis, and treatment resistance. We analyzed the activity of the proteasome, the primary organelle for targeted protein degradation, as a marker of tumor- and metastasis-initiating cells. Using human and mouse breast cancer cells expressing a validated fluorescent reporter, we found a small subpopulation of cells with low proteasome activity that divided asymmetrically to produce daughter cells with low or high proteasome activity. Breast cancer cells with low proteasome activity had greater local tumor formation and metastasis in immunocompromised and immunocompetent mice. To allow flexible labeling of cells, we also developed a new proteasome substrate based on HaloTag technology. Patient-derived glioblastoma cells with low proteasome activity measured by the HaloTag reporter show key phenotypes associated with tumor-initiating cells, including expression of a stem cell transcription factor, reconstitution of the original starting population, and enhanced neurosphere formation. We also show that patient-derived glioblastoma cells with low proteasome activity have higher frequency of tumor formation in mouse xenografts. These studies support proteasome function as a tool to investigate tumor- and metastasis-initiating cancer cells and a potential biomarker for outcomes in patients with several different cancers.

  19. Assessing Subunit Dependency of the Plasmodium Proteasome Using Small Molecule Inhibitors and Active Site Probes

    NARCIS (Netherlands)

    Li, H.; Linden, W.A. van der; Verdoes, M.; Florea, B.I.; McAllister, F.E.; Govindaswamy, K.; Elias, J.E.; Bhanot, P.; Overkleeft, H.S.; Bogyo, M.

    2014-01-01

    The ubiquitin-proteasome system (UPS) is a potential pathway for therapeutic intervention for pathogens such as Plasmodium, the causative agent of malaria. However, due to the essential nature of this proteolytic pathway, proteasome inhibitors must avoid inhibition of the host enzyme complex to

  20. Nelfinavir augments proteasome inhibition by bortezomib in myeloma cells and overcomes bortezomib and carfilzomib resistance

    International Nuclear Information System (INIS)

    Kraus, M; Bader, J; Overkleeft, H; Driessen, C

    2013-01-01

    HIV protease inhibitors (HIV-PI) are oral drugs for HIV treatment. HIV-PI have antitumor activity via induction of ER-stress, inhibition of phospho-AKT (p-AKT) and the proteasome, suggesting antimyeloma activity. We characterize the effects of all approved HIV-PI on myeloma cells. HIV-PI were compared regarding cytotoxicity, proteasome activity, ER-stress induction and AKT phosphorylation using myeloma cells in vitro. Nelfinavir is the HIV-PI with highest cytotoxic activity against primary myeloma cells and with an IC 50 near therapeutic drug blood levels (8–14 μM), irrespective of bortezomib sensitivity. Only nelfinavir inhibited intracellular proteasome activity in situ at drug concentrations <40 μℳ. Ritonavir, saquinavir and lopinavir inhibited p-AKT comparable to nelfinavir, and showed similar synergistic cytotoxicity with bortezomib against bortezomib-sensitive cells. Nelfinavir had superior synergistic activity with bortezomib/carfilzomib in particular against bortezomib/carfilzomib-resistant myeloma cells. It inhibited not only the proteasomal β1/β5 active sites, similar to bortezomib/carfilzomib, but in addition the β2 proteasome activity not targeted by bortezomib/carfilzomib. Additional inhibition of β2 proteasome activity is known to sensitize cells for bortezomib and carfilzomib. Nelfinavir has unique proteasome inhibiting activity in particular on the bortezomib/carfilzomib-insensitive tryptic (β2) proteasome activity in intact myeloma cells, and is active against bortezomib/carfilzomib-resistant myeloma cells in vitro

  1. Modulation of the Proteasome Pathway by Nano-Curcumin and Curcumin in Retinal Pigment Epithelial Cells

    NARCIS (Netherlands)

    Ramos de Carvalho, J. Emanuel; Verwoert, Milan T.; Vogels, Ilse M. C.; Schipper-Krom, Sabine; van Noorden, Cornelis J. F.; Reits, Eric A.; Klaassen, Ingeborg; Schlingemann, Reinier O.

    2018-01-01

    Curcumin has multiple biological effects including the modulation of protein homeostasis by the ubiquitin-proteasome system. The purpose of this study was to assess the in vitro cytotoxic and oxidative effects of nano-curcumin and standard curcumin and characterize their effects on proteasome

  2. High-throughput siRNA screening applied to the ubiquitin-proteasome system

    DEFF Research Database (Denmark)

    Poulsen, Esben Guldahl; Nielsen, Sofie V.; Pietras, Elin J.

    2016-01-01

    The ubiquitin-proteasome system is the major pathway for intracellular protein degradation in eukaryotic cells. Due to the large number of genes dedicated to the ubiquitin-proteasome system, mapping degradation pathways for short lived proteins is a daunting task, in particular in mammalian cells...

  3. Characterization of the proteasome from the extremely halophilic archaeon Haloarcula marismortui

    Directory of Open Access Journals (Sweden)

    B. Franzetti

    2002-01-01

    Full Text Available A 20S proteasome, comprising two subunits α and β, was purified from the extreme halophilic archaeon Haloarcula marismortui, which grows only in saturated salt conditions. The three-dimensional reconstruction of the H. marismortui proteasome (Hm proteasome, obtained from negatively stained electron micrographs, is virtually identical to the structure of a thermophilic proteasome filtered to the same resolution. The stability of the Hm proteasome was found to be less salt-dependent than that of other halophilic enzymes previously described. The proteolytic activity of the Hm proteasome was investigated using the malate dehydrogenase from H. marismortui (HmMalDH as a model substrate. The HmMalDH denatures when the salt concentration is decreased below 2 M. Under these conditions, the proteasome efficiently cleaves HmMalDH during its denaturation process, but the fully denatured HmMalDH is poorly degraded. These in vitro experiments show that, at low salt concentrations, the 20S proteasome from halophilic archaea eliminates a misfolded protein.

  4. Ciliopathy proteins regulate paracrine signaling by modulating proteasomal degradation of mediators

    Science.gov (United States)

    Liu, Yangfan P.; Tsai, I-Chun; Morleo, Manuela; Oh, Edwin C.; Leitch, Carmen C.; Massa, Filomena; Lee, Byung-Hoon; Parker, David S.; Finley, Daniel; Zaghloul, Norann A.; Franco, Brunella; Katsanis, Nicholas

    2014-01-01

    Cilia are critical mediators of paracrine signaling; however, it is unknown whether proteins that contribute to ciliopathies converge on multiple paracrine pathways through a common mechanism. Here, we show that loss of cilopathy-associated proteins Bardet-Biedl syndrome 4 (BBS4) or oral-facial-digital syndrome 1 (OFD1) results in the accumulation of signaling mediators normally targeted for proteasomal degradation. In WT cells, several BBS proteins and OFD1 interacted with proteasomal subunits, and loss of either BBS4 or OFD1 led to depletion of multiple subunits from the centrosomal proteasome. Furthermore, overexpression of proteasomal regulatory components or treatment with proteasomal activators sulforaphane (SFN) and mevalonolactone (MVA) ameliorated signaling defects in cells lacking BBS1, BBS4, and OFD1, in morphant zebrafish embryos, and in induced neurons from Ofd1-deficient mice. Finally, we tested the hypothesis that other proteasome-dependent pathways not known to be associated with ciliopathies are defective in the absence of ciliopathy proteins. We found that loss of BBS1, BBS4, or OFD1 led to decreased NF-κB activity and concomitant IκBβ accumulation and that these defects were ameliorated with SFN treatment. Taken together, our data indicate that basal body proteasomal regulation governs paracrine signaling pathways and suggest that augmenting proteasomal function might benefit ciliopathy patients. PMID:24691443

  5. Mouse zygote-specific proteasome assembly chaperone important for maternal-to-zygotic transition

    Directory of Open Access Journals (Sweden)

    Seung-Wook Shin

    2012-11-01

    During the maternal-to-zygotic transition (MZT, maternal proteins in oocytes are degraded by the ubiquitin–proteasome system (UPS, and new proteins are synthesized from the zygotic genome. However, the specific mechanisms underlying the UPS at the MZT are not well understood. We identified a molecule named zygote-specific proteasome assembly chaperone (ZPAC that is specifically expressed in mouse gonads, and expression of ZPAC was transiently increased at the mouse MZT. ZPAC formed a complex with Ump1 and associated with precursor forms of 20S proteasomes. Transcription of ZPAC genes was also under the control of an autoregulatory feedback mechanism for the compensation of reduced proteasome activity similar to Ump1 and 20S proteasome subunit gene expression. Knockdown of ZPAC in early embryos caused a significant reduction of proteasome activity and decrease in Ump1 and mature proteasomes, leading to accumulation of proteins that need to be degraded at the MZT and early developmental arrest. Therefore, a unique proteasome assembly pathway mediated by ZPAC is important for progression of the mouse MZT.

  6. BAG3 induces the sequestration of proteasomal clients into cytoplasmic puncta

    Science.gov (United States)

    Minoia, Melania; Boncoraglio, Alessandra; Vinet, Jonathan; Morelli, Federica F; Brunsting, Jeanette F; Poletti, Angelo; Krom, Sabine; Reits, Eric; Kampinga, Harm H; Carra, Serena

    2014-01-01

    Eukaryotic cells use autophagy and the ubiquitin–proteasome system as their major protein degradation pathways. Upon proteasomal impairment, cells switch to autophagy to ensure proper clearance of clients (the proteasome-to-autophagy switch). The HSPA8 and HSPA1A cochaperone BAG3 has been suggested to be involved in this switch. However, at present it is still unknown whether and to what extent BAG3 can indeed reroute proteasomal clients to the autophagosomal pathway. Here, we show that BAG3 induces the sequestration of ubiquitinated clients into cytoplasmic puncta colabeled with canonical autophagy linkers and markers. Following proteasome inhibition, BAG3 upregulation significantly contributes to the compensatory activation of autophagy and to the degradation of the (poly)ubiquitinated proteins. BAG3 binding to the ubiquitinated clients occurs through the BAG domain, in competition with BAG1, another BAG family member, that normally directs ubiquitinated clients to the proteasome. Therefore, we propose that following proteasome impairment, increasing the BAG3/BAG1 ratio ensures the “BAG-instructed proteasomal to autophagosomal switch and sorting” (BIPASS). PMID:25046115

  7. The effects of Hot Pepper Extract and Capsaicin on Adipocyte Metabolism

    Directory of Open Access Journals (Sweden)

    Ching Sheng, Chu

    2008-03-01

    Full Text Available Objectives : The purpose of this study is to investigate the effects of hot pepper extract and capsaicin on the adipogenesis in 3T3-L1 cells, lipolysis in rat epididymal adipocytes and histological changes in porcine adipose tissue. Methods : Inhibiton of preadipocyte differentiation and/or stimulation of lipolysis play important roles in reducing obesity. 3T3-L1 preadipocytes were differentiated with adipogenic reagents by incubating for 3 days in the absence or presence of hot pepper extract or capsaicin ranging from 0.01 to 1㎎/㎖. The effects of hot pepper extract and capsaicin on adipogenesis were examined by measuring GPDH activity and by Oil Red O staining. Mature adipocytes from rat epididymal fat pad was incubated with hot pepper extract or capsaicin ranging from 0.01 to 1㎎/㎖ for 3 hrs. The effects of hot pepper extract and capsaicin on lipolysis were examined by measuring free glycerol released. Fat tissue from pig skin was injected with hot pepper extract or capsaicinCFP ranging from 0.1 to 10㎎/㎖ to examine the effects of hot pepper extract and capsaicin on histological changes under light microscopy. Results : The following results were obtained from present study on adipogenesis of preadipocytes, lipolysis of adipocytes and histological changes in fat tissue. 1. Hot pepper extract and capsaicin inhibited adipogenic differentiation at the concentration of 0.1 and 0.01㎎/㎖, respectively, indicating that capsaicin was more effective in inhibiting adipogenesis than hot pepper extract. 2. Hot pepper extract and capsaicin decreased the activity of glycerol-3-phosphate dehydrogenase(GPDH at the concentration of 0.1 and 0.01㎎/㎖, respectively, indicating that capsaicin was more effective in inhibiting adipogenic differentiation than hot pepper extract. 3. Hot pepper extract and capsaicin increased glycerol release at the concentration of 0.1㎎/㎖. There was no difference in lipolytic activity between hot pepper extract and

  8. Distinct temporal requirements for autophagy and the proteasome in yeast meiosis.

    Science.gov (United States)

    Wen, Fu-ping; Guo, Yue-shuai; Hu, Yang; Liu, Wei-xiao; Wang, Qian; Wang, Yuan-ting; Yu, Hai-Yan; Tang, Chao-ming; Yang, Jun; Zhou, Tao; Xie, Zhi-ping; Sha, Jia-hao; Guo, Xuejiang; Li, Wei

    2016-01-01

    Meiosis is a special type of cellular renovation that involves 2 successive cell divisions and a single round of DNA replication. Two major degradation systems, the autophagy-lysosome and the ubiquitin-proteasome, are involved in meiosis, but their roles have yet to be elucidated. Here we show that autophagy mainly affects the initiation of meiosis but not the nuclear division. Autophagy works not only by serving as a dynamic recycling system but also by eliminating some negative meiotic regulators such as Ego4 (Ynr034w-a). In a quantitative proteomics study, the proteasome was found to be significantly upregulated during meiotic divisions. We found that proteasomal activity is essential to the 2 successive meiotic nuclear divisions but not for the initiation of meiosis. Our study defines the roles of autophagy and the proteasome in meiosis: Autophagy mainly affects the initiation of meiosis, whereas the proteasome mainly affects the 2 successive meiotic divisions.

  9. Proteasomal Dysfunction Induced By Diclofenac Engenders Apoptosis Through Mitochondrial Pathway.

    Science.gov (United States)

    Amanullah, Ayeman; Upadhyay, Arun; Chhangani, Deepak; Joshi, Vibhuti; Mishra, Ribhav; Yamanaka, Koji; Mishra, Amit

    2017-05-01

    Diclofenac is the most commonly used phenylacetic acid derivative non-steroidal anti-inflammatory drug (NSAID) that demonstrates significant analgesic, antipyretic, and anti-inflammatory effects. Several epidemiological studies have demonstrated anti-proliferative activity of NSAIDs and examined their apoptotic induction effects in different cancer cell lines. However, the precise molecular mechanisms by which these pharmacological agents induce apoptosis and exert anti-carcinogenic properties are not well known. Here, we have observed that diclofenac treatment induces proteasome malfunction and promotes accumulation of different critical proteasome substrates, including few pro-apoptotic proteins in cells. Exposure of diclofenac consequently elevates aggregation of various ubiquitylated misfolded proteins. Finally, we have shown that diclofenac treatment promotes apoptosis in cells, which could be because of mitochondrial membrane depolarization and cytochrome c release into cytosol. This study suggests possible beneficial insights of NSAIDs-induced apoptosis that may improve our existing knowledge in anti-proliferative interspecific strategies development. J. Cell. Biochem. 118: 1014-1027, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  10. Improvement of hygienic quality and long-term storage of dried red pepper by gamma irradiation

    International Nuclear Information System (INIS)

    Byun, M.W.; Yook, H.S.; Kwon, J.H.; Kim, J.O.

    1996-01-01

    Dried-red pepper, whole and powdered types, was subjected to a storage-study by investigation the effects of packaging methods (polycloth & polyethylene/polycloth, whole dried-red pepper; nylon/polyethylene-lam-inated film, red pepper powder), temperature and gamma irradiation doses (0-10 kGy). After 6 months storage in polyclith sack at ambient temperature, all whole dried-red pepper showed quality deterioration, such as weight change, insect infestation, discoloration and chemical changes, After 2 years storage in combined packaging with polyethylene/polycloth sack of 5-7.5 kGy irradiated whole dried-red pepper at ambient temperature, however, quality deterioration was not observed. Gamma-irradiated red pepper powder (7.5-10kGy) showed a good quality in hygienic, physicochemical and organoleptic evaluation after 2 years of storage at ambient temperature

  11. Key Microbiota Identification Using Functional Gene Analysis during Pepper (Piper nigrum L.) Peeling.

    Science.gov (United States)

    Zhang, Jiachao; Hu, Qisong; Xu, Chuanbiao; Liu, Sixin; Li, Congfa

    2016-01-01

    Pepper pericarp microbiota plays an important role in the pepper peeling process for the production of white pepper. We collected pepper samples at different peeling time points from Hainan Province, China, and used a metagenomic approach to identify changes in the pericarp microbiota based on functional gene analysis. UniFrac distance-based principal coordinates analysis revealed significant changes in the pericarp microbiota structure during peeling, which were attributed to increases in bacteria from the genera Selenomonas and Prevotella. We identified 28 core operational taxonomic units at each time point, mainly belonging to Selenomonas, Prevotella, Megasphaera, Anaerovibrio, and Clostridium genera. The results were confirmed by quantitative polymerase chain reaction. At the functional level, we observed significant increases in microbial features related to acetyl xylan esterase and pectinesterase for pericarp degradation during peeling. These findings offer a new insight into biodegradation for pepper peeling and will promote the development of the white pepper industry.

  12. Relaxation behavior of radicals produced in irradiated black pepper under various moisture conditions by ESR

    International Nuclear Information System (INIS)

    Kameya, Hiromi; Kawauchi, Risa; Shimoyama, Yuhei; Kaeda, Yoko; Ogawa, Satoko; Nakamura, Hideo; Ukai, Mitsuko

    2008-01-01

    Black pepper is easy to be contaminated by microorganism and often processed to γ-irradiation. ESR has been used for the detection of radicals induced in irradiated spices. Using ESR, we revealed the effects of moisture condition during storage of irradiated black pepper on the saturation behavior of ESR signal. The ESR spectrum of black pepper consists of a broad sextet centered at g=2.0, a singlet as same g-value and a singlet at g=4.0. The irradiation causes two new signals, one is the strong and sharp singlet signal at g=2.0 and the other is the side signal. We found that the signal intensity originated by the radicals of black pepper with and without radiation decayed in the high humidity condition during storage. The ESR signal intensity of irradiated black pepper decayed during storage and showed almost the same intensity level as that of non-irradiated black pepper during storage. (author)

  13. Identification of irradiated peppers by electron spin resonance, thermoluminescence and viscosity

    International Nuclear Information System (INIS)

    Polonia, I.; Esteves, M.P.; Andrade, M.E.; Laboratorio Nacional de Engenharia e Tecnologia Industrial, Sacavem; Empis, J.

    1995-01-01

    White and black pepper purchased in local retailers were analysed by electron spin resonance (ESR), thermoluminescence (TL) and viscosimetry (VISC) in order to establish a viable method for identifying possibly irradiated peppers. Samples studied were non irradiated or irradiated in a cobalt-60 plant with the absorbed doses of 3, 5, 7 and 10 kGy. Confirming the data found in the literature TL was revealed by our results the best method to identify irradiated peppers. Nevertheless, the dose received by the samples could not be estimated. The ESR signal of irradiated peppers is similar to the spectrum of cellulose radical but very short lived at ambient temperature. The study on the alteration of viscosity of heat-treated alkaline pepper suspensions indicate that VISC is a very promising method for detection of irradiated peppers. (Author)

  14. Antioxidant, Antinociceptive, and Anti-Inflammatory Effects of Carotenoids Extracted from Dried Pepper (Capsicum annuum L.)

    Science.gov (United States)

    Hernández-Ortega, Marcela; Ortiz-Moreno, Alicia; Hernández-Navarro, María Dolores; Chamorro-Cevallos, Germán; Dorantes-Alvarez, Lidia; Necoechea-Mondragón, Hugo

    2012-01-01

    Carotenoids extracted from dried peppers were evaluated for their antioxidant, analgesic, and anti-inflammatory activities. Peppers had a substantial carotenoid content: guajillo 3406 ± 4 μg/g, pasilla 2933 ± 1 μg/g, and ancho 1437 ± 6 μg/g of sample in dry weight basis. A complex mixture of carotenoids was discovered in each pepper extract. The TLC analysis revealed the presence of chlorophylls in the pigment extract from pasilla and ancho peppers. Guajillo pepper carotenoid extracts exhibited good antioxidant activity and had the best scavenging capacity for the DPPH+ cation (24.2%). They also exhibited significant peripheral analgesic activity at 5, 20, and 80 mg/kg and induced central analgesia at 80 mg/kg. The results suggest that the carotenoids in dried guajillo peppers have significant analgesic and anti-inflammatory benefits and could be useful for pain and inflammation relief. PMID:23091348

  15. Detection of irradiated peppers by viscosity measurement at extremely high pH

    International Nuclear Information System (INIS)

    Hayashi, Toru; Todoriki, Setsuko

    1996-01-01

    The viscosities of aqueous suspensions of irradiated peppers determined after heat gelatinization were influenced by the pH of the suspension to a greater degree than those of unirradiated ones. Viscosity measurement under an extremely alkaline condition (pH 13.8) resulted in a significant different between irradiated peppers and unirradiated ones, irrespective of the planting locality and storage period. All of the pepper samples irradiated at 5 kGy showed viscosity values significantly lower than unirradiated ones. (Author)

  16. Mixed noble gas effect on cut green peppers

    Science.gov (United States)

    Raymond, L. V.; Zhang, M.; Karangwa, E.; Chesereka, M. J.

    2013-01-01

    Increasing attempts at using gas which leads to hydrate formation as a preservative tool in fresh-cut fruits and vegetables have been reported. In this study, changes in some physical and biochemical properties of fresh-cut green peppers under compressed noble gas treatments were examined. Mixed argonkrypton and argon treatments were performed before cold storage at 5°C for 15 days. Mass loss and cell membrane permeability were found to be the lowest in mixed argon-krypton samples. Besides, a lower CO2 concentration and vitamin C loss were detected in gastreated samples compared to untreated samples (control). While the total phenol degradation was moderately reduced, the effect of the treatment on polyphenoloxidase activity was better at the beginning of the storage period. The minimum changes in quality observed in cut peppers resulted from both mixed and gas treatment alone.

  17. Isolation of microorganisms from red pepper powder and their radiosensitivity

    Energy Technology Data Exchange (ETDEWEB)

    Choi, E H; Kim, Y B; Lee, S R [Korea Atomic Energy Research Inst., Seoul (Republic of Korea)

    1977-01-01

    From samples of red pepper powder sold in Korea were isolated and identified 13 species of molds (Aspergillus amsteodami, Asp. chevalieri, Asp. clavatus, Asp. Flavus, Asp. janus var. effusus, Asp. oryzae, Asp. oryzae var. brevis, Asp. repens, Asp. sydowi, Asp. thomii, Asp. tubingensis, Penicillium thomii, Scopulariopsis brevicaulis) and 5 species of bacteria (Bacillus pumilus, Bac. subtilis, Micrococus luteus, M. varians, Staphylococcus aureus). Radiosensitivity of these microorganisms was examined to give D/sub 10/ values of 14-41 krad for molds, 11-24 krad for bacterial vegetative cells and 190-250 krad for bacterial spores. The red pepper powder was contaminated with 2-3x10/sup 2/ mold counts/g and 3-6x10/sup 7/ bacterial counts/g, which would be sufficiently destroyed by irradiating 200 krad r-rays.

  18. Storage stability and irradiation effect of red pepper powder

    Energy Technology Data Exchange (ETDEWEB)

    Lee, C H; Choi, E H; Lee, S R [Korea Atomic Energy Research Inst., Seoul (Republic of Korea); Kim, H S [Yonsei Univ., Seoul (Republic of Korea)

    1977-01-01

    To improve the storage stability of Korea red pepper (Capsicum annuum), the equilibrium moisture content of the powder and effects of irradiation on the capsicin, capsanthin and microorganisms were investigated during its storage. The equilibrium moisture contents of the powder at 25 /sup 0/C and relative humidities of 43.7, 53.3, 64.4, 75.4, and 84.3% were 13, 18, 25, 28, and 37%, respectively, and the larger the particle size was, the lower the equilibrium moisture content was under the same storage conditions. Cobalt-60 gamma irradiation degraded partly the capsaicin in red pepper or in solution at dose levels above 500 krad, but did not affect the moisture and capsanthin contents during its storage. Total visible cell count of the powder was around 10/sup 8/ per gram and D/sub 10/ value for the mixed organisms was 210 krad.

  19. Slit scattering effects in a well aligned pepper pot

    International Nuclear Information System (INIS)

    Power, J. G.

    2003-01-01

    A pepper pot is a device used to measure a medium energy (< 20 MeV) electron beam's transverse emittance by sampling its transverse phase space. This is accomplished by blocking most of the incident electron beam, while allowing small 'beamlets' to pass through openings in a mask. The accuracy of the transverse emittance measured by a pepper pot is limited by several factors including, electrons leaking through the solid region of the mask, the imaging system resolution and dynamic range, scattering, etc. While the noise contributions from the prior quantities can be easily estimated, scattering effects have previously been neglected due to the difficulty in estimating the effect. In this paper, EGS4 simulations are presented to determine the affect of scattering on emittance measurements for an 8 MeV electron beam

  20. Vacuum-assisted microwave drying characteristics of green bell pepper

    Directory of Open Access Journals (Sweden)

    Vivek Kumar

    2017-04-01

    Full Text Available Chopped green bell pepper pieces were blanched (95 °C, 5 min and chemically pretreated (1% potassium metabisulphite solution, 25 min at room temperature before drying in hot air dryer (HAD at various temperature ranges (60 – 80 °C. Three vacuum levels (200, 400, 600 mm Hg and microwave power levels (100, 200, 300 W were also used to dry green bell pepper samples in a vacuum assisted microwave (VAM (2.45 GHz, 0.8 kW dryer. VAM drying methods offered a maximum reduction by four to five times in drying time as compared to that in HAD. The logarithmic model was found to have the best fit based on high R2 and small values of reduced χ2 and RMSE.  VAM method has higher values for effective moisture diffusivity (Deff and lower values for activation energy (Ea, in comparison to the HAD method. 

  1. Isolation of microorganisms from red pepper powder and their radiosensitivity

    International Nuclear Information System (INIS)

    Choi, E.H.; Kim, Y.B.; Lee, S.R.

    1977-01-01

    From samples of red pepper powder sold in Korea were isolated and identified 13 species of molds (Aspergillus amsteodami, Asp. chevalieri, Asp. clavatus, Asp. Flavus, Asp. janus var. effusus, Asp. oryzae, Asp. oryzae var. brevis, Asp. repens, Asp. sydowi, Asp. thomii, Asp. tubingensis, Penicillium thomii, Scopulariopsis brevicaulis) and 5 species of bacteria (Bacillus pumilus, Bac. subtilis, Micrococus luteus, M. varians, Staphylococcus aureus). Radiosensitivity of these microorganisms was examined to give D 10 values of 14-41 krad for molds, 11-24 krad for bacterial vegetative cells and 190-250 krad for bacterial spores. The red pepper powder was contaminated with 2-3x10 2 mold counts/g and 3-6x10 7 bacterial counts/g, which would be sufficiently destroyed by irradiating 200 krad r-rays. (Author)

  2. The Dynamic Microbiota Profile During Pepper (Piper nigrum L.) Peeling by Solid-State Fermentation.

    Science.gov (United States)

    Hu, Qisong; Zhang, Jiachao; Xu, Chuanbiao; Li, Congfa; Liu, Sixin

    2017-06-01

    White pepper (Piper nigrum L.), a well-known spice, is the main pepper processing product in Hainan province, China. The solid-state method of fermentation can peel pepper in a highly efficient manner and yield high-quality white pepper. In the present study, we used next-generation sequencing to reveal the dynamic changes in the microbiota during pepper peeling by solid-state fermentation. The results suggested that the inoculated Aspergillus niger was dominant throughout the fermentation stage, with its strains constituting more than 95% of the fungi present; thus, the fungal community structure was relatively stable. The bacterial community structure fluctuated across different fermentation periods; among the bacteria present, Pseudomonas, Tatumella, Pantoea, Acinetobacter, Lactococcus, and Enterobacter accounted for more than 95% of all bacteria. Based on the correlations among the microbial community, we found that Pseudomonas and Acinetobacter were significantly positively related with A. niger, which showed strong synergy with them. In view of the microbial functional gene analysis, we found that these three bacteria and fungi were closely related to the production of pectin esterase (COG4677) and acetyl xylan esterase (COG3458), the key enzymes for pepper peeling. The present research clarifies the solid-state fermentation method of pepper peeling and lays a theoretical foundation to promote the development of the pepper peeling process and the production of high-quality white pepper.

  3. Influence of the heat treatment on the color of ground pepper (Capsicum annuum

    Directory of Open Access Journals (Sweden)

    Vračar Ljubo O.

    2007-01-01

    Full Text Available Red pepper (Capsicum annuum L. is one of the most important vegetables in the world. The main ground pepper quality attributes are extractable color, surface color, qualitative and quantitative carotenoid content. In this work, the influence of heat treatment on ground pepper quality was investigated. Microbiological status was examined in non-sterilized and sterilized ground pepper. Color changes were assessed by measuring the extractable color (ASTA and surface color, using a photocolorimeter. The obtained results showed that at the end of experiment, non-sterilized samples had higher color values in comparison to the sterilized ones. Also, color deterioration was heightened at room temperature.

  4. Preliminary examination of induced radioactivity in pepper by 10 MeV electron irradiation

    International Nuclear Information System (INIS)

    Katayama, Tadashi; Furuta, Masakazu; Sibata, Setsuko; Ito, Norio; Mizohata, Akira; Matsunami, Tadao; Toratani, Hirokazu; Takeda, Atsuhiko.

    1991-01-01

    β-ray measurement was performed on 10 MeV electron-irradiated black pepper and white pepper with liquid scintillation counter in order to reconfirm the wholesomeness of irradiated foods and present unambiguous data to general consumers concerning about the induced radioactivity in the irradiated foods. In irradiated black pepper no radioactivity other than from natural source, un-irradiated one, was detected. But in irradiated white pepper, it was suggested that induced radioactivity might be detected if the detection method was more improved. (author)

  5. The decrease in aqueous tear production associated with pepper spray.

    Science.gov (United States)

    Rasier, Rifat; Kukner, Amber Senel; Sengul, Elvan Alper; Yalcin, Nazli Gul; Temizsoylu, Onur; Bahcecioglu, Halil Onder

    2015-04-01

    Pepper spray is used both by civilians and by law enforcement. Burning sensation occurs when exposed to skin, pain and temporary blindness occurs when exposed to the eyes. This study focused on the effect of pepper spray on lacrimal tear production and subsequently on corneal sensitivity in a large group after an intense exposure. Ninety-six people who were exposed to pepper spray during the Gezi park protests volunteered. Subjects were asked if they wore any protective goggles and if they irrigated their eyes after exposure. They were asked to record their symptoms regarding dry eye in a standardized questionnaire. Schirmer I and II tests were performed. Eighty-two people wore protective goggles during exposure, whereas 14 people did not have any protection. Both Schirmer results in unprotected subjects were significantly lower than that in protected subjects. Schirmer I and II results of unprotected subjects were not statistically different, whereas they were statistically different in protected subjects. Thirty-five percent of unprotected subjects and 24% of protected subjects expressed symptoms of dry eye. The active ingredient of pepper spray is oleoresin capsicum. It is randomly diffused to polymodal nerve terminals, leading to opening of non-selective cationic channels and block neuronal transmission. The lower results of both Schirmer in unprotected group emphasize the importance of a protective Google glass around the eyes during exposal, serving as a barrier minimalizing the contact of the spray with the eyes. The combination of the low results and lack of symptoms could suggest that corneal reflex lacrimation in our subjects was not abundant enough. The findings of this study could not fully represent long term findings but it could be assumed that our findings could be indicative of the sensory denervation and alterations demonstrated in studies investigating the long term effects of oleoresin capsicum.

  6. Recent approaches for bell pepper seed vigor testing.

    OpenAIRE

    DIAS, M. A. N.; MONDO, V. H. V.; CICERO, S. M.

    2014-01-01

    Seed vigor is a major attribute in the determination of whether a seed lot can be commercialized. Different physiological tests are used to assess seed vigor and are generally based on different concepts, which could lead to variable interpretations, particularly when comparing lots presenting different vigor levels. This study was conducted to compare the different approaches used to evaluate bell pepper seed vigor and verify the applicability of the Seed Vigor Imaging System (SVIS) for this...

  7. Recent approaches for bell pepper seed vigor testing

    OpenAIRE

    Dias, Marcos Altomani Neves; Mondo, Vitor Henrique Vaz; Cicero, Silvio Moure

    2014-01-01

    Seed vigor is a major attribute in the determination of whether a seed lot can be commercialized. Different physiological tests are used to assess seed vigor and are generally based on different concepts, which could lead to variable interpretations, particularly when comparing lots presenting different vigor levels. This study was conducted to compare the different approaches used to evaluate bell pepper seed vigor and verify the applicability of the Seed Vigor Imaging System (SVIS) for this...

  8. Detection and quantification of Leveillula taurica growth in pepper leaves.

    Science.gov (United States)

    Zheng, Zheng; Nonomura, Teruo; Bóka, Károly; Matsuda, Yoshinori; Visser, Richard G F; Toyoda, Hideyoshi; Kiss, Levente; Bai, Yuling

    2013-06-01

    Leveillula taurica is an obligate fungal pathogen that causes powdery mildew disease on a broad range of plants, including important crops such as pepper, tomato, eggplant, onion, cotton, and so on. The early stage of this disease is difficult to diagnose and the disease can easily spread unobserved; for example, in pepper and tomato production fields and greenhouses. The objective of this study was to develop a detection and quantification method of L. taurica biomass in pepper leaves with special regard to the early stages of infection. We monitored the development of the disease to time the infection process on the leaf surface as well as inside the pepper leaves. The initial and final steps of the infection taking place on the leaf surface were consecutively observed using a dissecting microscope and a scanning electron microscope. The development of the intercellular mycelium in the mesophyll was followed by light and transmission electron microscopy. A pair of L. taurica-specific primers was designed based on the internal transcribed spacer sequence of L. taurica and used in real-time polymerase chain reaction (PCR) assay to quantify the fungal DNA during infection. The specificity of this assay was confirmed by testing the primer pair with DNA from host plants and also from another powdery mildew species, Oidium neolycopersici, infecting tomato. A standard curve was obtained for absolute quantification of L. taurica biomass. In addition, we tested a relative quantification method by using a plant gene as reference and the obtained results were compared with the visual disease index scoring. The real-time PCR assay for L. taurica provides a valuable tool for detection and quantification of this pathogen in breeding activities as well in plant-microbe interaction studies.

  9. Characterization of a pepper collection (Capsicum frutescens L.) from Brazil.

    Science.gov (United States)

    Lima, M F; Carvalho, S I C; Ragassi, C F; Bianchetti, L B; Faleiro, F G; Reifschneider, F J B

    2017-08-31

    Germplasm banks are essential as sources of genetic variability for plant breeding programs. To characterize a Brazilian Capsicum frutescens collection, 21 malagueta and 5 Tabasco hot pepper accessions were evaluated under field and greenhouse conditions regarding morphological and molecular traits, as well as resistance to viruses. Morphological characterization was performed using 53 IPGRI (International Plant Genetic Resources Institute) descriptors, 15 vegetative, 13 inflorescence, 22 fruit, and 3 seed. Molecular characterization was carried out with 60 polymorphic markers from 29 RAPD primers. The incidence of major viruses infecting Capsicum spp, Tomato spotted wilt virus (TSWV), Groundnut ringspot virus (GRSV), Potato virus Y (PVY), Pepper yellow mosaic virus (PepYMV), Pepper mild mottle virus (PMMoV), and Cucumber mosaic virus (CMV) was evaluated by ELISA. Based on the average genetic distance among genotypes, six groups were defined for the 53 IPGRI descriptors. When considering only 11 quantitative traits (five vegetative and six fruit), six groups were also determined, and the traits plant canopy width (56.05%) and days to fruiting (25.07%) most explained the genetic diversity among genotypes. Molecular analysis defined five groups of accessions with partial correspondence to the morphological characterization data. The incidence of viruses in field-grown plants varied among genotypes and according to virus species, from 5.6% (GRSV; CNPH 3286) to 100% (PMMoV; CNPH2871), and indicated some accessions as potential sources of virus resistance. These results demonstrate the genetic variability within the group of 26 hot pepper accessions, as well as virus-resistant genotypes that can be used in breeding programs.

  10. Reaction of sweet pepper to the potato virus y (PVYm

    Directory of Open Access Journals (Sweden)

    Echer Márcia de Moraes

    2002-01-01

    Full Text Available Traditional sweet pepper cultivars showing susceptibility to the Potato virus Y (PVY are being replaced by resistant hybrids with higher commercial value. Despite of much information about resistance source reaction and their inheritance, there is no knowledge about the genetic background of commercial resistant hybrids. Reaction of sweet pepper (Capsicum annuum L. hybrids to the Potato virus Y (PVYm such as Acuario, Magali R, Nathalie and their respective generations F2 and F3 as well as hybrids Amanda, Corteso W208, CPC-6272, Dagmar, Elisa, Magali, Margarita, Monteiro, Quantum, Vivo W205 was evaluated. Reaction to PVYm was evaluated as resistant or susceptible. Magali R and Nathalie hybrid did not show any mosaic symptoms. Magali R and Nathalie hybrids resistance is due to a single dominant gene indicating resistant versus susceptible parental lines crossing pedigree. Amanda, Acuario, Corteso W208, Dagmar, Elisa, Margarita, Monteiro, Quantum and Vivo W205, considered resistant to PVY, were highly susceptibility to PVY strain m. Hybrids, claimed as resistant to the Pepper mottle virus (PepMoV, were also resistant to PVYm.

  11. Introducing Natural Farming in Black Pepper (Piper nigrum L. Cultivation

    Directory of Open Access Journals (Sweden)

    Kevin Muyang Tawie Sulok

    2018-01-01

    Full Text Available This paper reviews the role of Natural Farming as an ecological farming method to produce organically grown food of safe and high quality and at the same time improve soil quality and soil health. Currently, there is a dearth of information on the effects of Natural Farming approach on black pepper farms particularly in Sarawak, Malaysia. Previous studies on other crops had indicated positive outcome using the Natural Farming method. Thus, this paper discusses the essential role of effective microorganisms in Natural Farming and their potential in pepper cultivation. Through the action of effective microorganisms, this approach should be able to transform a degraded soil ecosystem into one that is fertile and has high nutrients availability. The mixed culture of effective microorganisms applied must be mutually compatible and coexist with one another to ensure its favorable establishment and interaction in the soil. Therefore, it is anticipated that introducing Natural Farming in black pepper cultivation can enhance the predominance of effective microorganisms in the soil, which in turn could lead to promising growth and yield of the crop.

  12. Tissue culture of black pepper (piper nigrum l.) in Pakistan

    International Nuclear Information System (INIS)

    Hussain, A.; Naz, S.; Nazir, H.; Shinwari, Z.K.

    2011-01-01

    Black pepper (Piper nigrum L.) the 'King of Spices' is a universal table condiment. It is extensively used in Pakistani cuisines and herbal medicines and imported in bulk from neighboring countries. The black pepper vine is generally cultivated by seed because other vegetative propagation methods are slow and time consuming. Therefore the tissue culture technique is considered more efficient and reliable method for rapid and mass propagation of this economically important plant. The present study was initiated to develop protocol for micro-propagation of black pepper vine. The stem, leaf and shoot tip explants from mature vine were cultured on MS medium supplemented with different concentrations of plant growth regulators (2,4-D, BA, IBA). Best callus was produced on MS medium with 1.5 mg/l BA by shoot tip explant. Shoot regeneration was excellent on MS medium with 0.5 mg/l BA. The plantlets formed were rooted best on 1.5 mg/l IBA. The rooted plants were transplanted in soil medium and acclimatized in growth room. The plants raised were test planted under the local conditions of Hattar. (author)

  13. Proteasome- and Ethanol-Dependent Regulation of HCV-Infection Pathogenesis

    Directory of Open Access Journals (Sweden)

    Natalia A. Osna

    2014-09-01

    Full Text Available This paper reviews the role of the catabolism of HCV and signaling proteins in HCV protection and the involvement of ethanol in HCV-proteasome interactions. HCV specifically infects hepatocytes, and intracellularly expressed HCV proteins generate oxidative stress, which is further exacerbated by heavy drinking. The proteasome is the principal proteolytic system in cells, and its activity is sensitive to the level of cellular oxidative stress. Not only host proteins, but some HCV proteins are degraded by the proteasome, which, in turn, controls HCV propagation and is crucial for the elimination of the virus. Ubiquitylation of HCV proteins usually leads to the prevention of HCV propagation, while accumulation of undegraded viral proteins in the nuclear compartment exacerbates infection pathogenesis. Proteasome activity also regulates both innate and adaptive immunity in HCV-infected cells. In addition, the proteasome/immunoproteasome is activated by interferons, which also induce “early” and “late” interferon-sensitive genes (ISGs with anti-viral properties. Cleaving viral proteins to peptides in professional immune antigen presenting cells and infected (“target” hepatocytes that express the MHC class I-antigenic peptide complex, the proteasome regulates the clearance of infected hepatocytes by the immune system. Alcohol exposure prevents peptide cleavage by generating metabolites that impair proteasome activity, thereby providing escape mechanisms that interfere with efficient viral clearance to promote the persistence of HCV-infection.

  14. SUMOylation of Blimp-1 promotes its proteasomal degradation.

    Science.gov (United States)

    Shimshon, Livnat; Michaeli, Avital; Hadar, Rivka; Nutt, Stephen L; David, Yael; Navon, Ami; Waisman, Ari; Tirosh, Boaz

    2011-08-04

    B lymphocyte induced maturation protein-1 (Blimp-1) is a transcription repressor of the Krueppel-like family. Blimp-1 plays important roles in developmental processes, such as of germ cells and hair follicle stem cells. In B lymphocytes Blimp-1 orchestrates the terminal differentiation into plasma cells. We discovered that Blimp-1 undergoes SUMOylation by SUMO-1. This SUMOylation is modulated by the SUMO protease SENP1. While Blimp-1 is relatively stable in 293T cells, a fusion with SUMO1 rendered it to rapid proteasomal degradation. Increase in SENP1 activity stabilized Blimp-1, while a decrease promoted its degradation. Our data indicate that SUMOylation of Blimp-1 regulates its intracellular stability. Copyright © 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  15. Observation of a H- Beam at the CERN Linac 4 Test Stand using a Pepper-pot

    CERN Document Server

    Delerue, N; Midttun, O; Scrivens, R; Tsesmelis, E

    2010-01-01

    Pepper-pot based transverse emittance measurement has the advantage of providing a fast (single shot) measurement with a relatively simple hardware. We report on the installation of a pepper-pot at the CERN Linac 4 test stand.

  16. Proteasome inhibitors induce apoptosis and reduce viral replication in primary effusion lymphoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Saji, Chiaki [Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo 060-0812 (Japan); Higashi, Chizuka; Niinaka, Yasufumi [Faculty of Medicine, University of Yamanashi, Chuoh-shi 409-3898 (Japan); Yamada, Koji [Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo 060-0812 (Japan); Noguchi, Kohji [Faculty of Pharmacy, Keio University, 1-5-30 Shiba-koen, Minato-ku, Tokyo 105-8512 (Japan); Fujimuro, Masahiro, E-mail: fuji2@mb.kyoto-phu.ac.jp [Department of Cell Biology, Kyoto Pharmaceutical University, Misasagi-Shichonocho 1, Yamashinaku, Kyoto 607-8412 (Japan)

    2011-12-02

    Highlights: Black-Right-Pointing-Pointer Constitutive NF-{kappa}B signaling is essential for the survival and growth of PEL cells. Black-Right-Pointing-Pointer NF-{kappa}B signaling is upregulated by the proteasome-dependent degradation of I{kappa}B{alpha}. Black-Right-Pointing-Pointer Proteasome inhibitors suppress NF-{kappa}B signaling and induce apoptosis in PEL cells through stabilization of I{kappa}B{alpha}. Black-Right-Pointing-Pointer Proteasome inhibitors suppress viral replication in PEL cells during lytic KSHV infection. -- Abstract: Primary effusion lymphoma (PEL) is an aggressive neoplasm caused by Kaposi's sarcoma-associated herpesvirus (KSHV). This study provides evidence that proteasomal activity is required for both survival of PEL cells stably harboring the KSHV genome and viral replication of KSHV. We evaluated the cytotoxic effects of proteasome inhibitors on PEL cells. The proteasome inhibitors MG132, lactacystin, and proteasome inhibitor I dramatically inhibited cell proliferation and induced apoptosis of PEL cells through the accumulation of p21 and p27. Furthermore, proteasome inhibitors induced the stabilization of NF-{kappa}B inhibitory molecule (I{kappa}B{alpha}) and suppressed the transcriptional activity of NF-{kappa}B in PEL cells. The NF-{kappa}B specific inhibitor BAY11-7082 also induced apoptosis in PEL cells. The constitutive activation of NF-{kappa}B signaling is essential for the survival and growth of B cell lymphoma cells, including PEL cells. NF-{kappa}B signaling is upregulated by proteasome-dependent degradation of I{kappa}B{alpha}. The suppression of NF-{kappa}B signaling by proteasome inhibitors may contribute to the induction of apoptosis in PEL cells. In addition, proteasome activity is required for KSHV replication in KSHV latently infected PEL cells. MG132 reduced the production of progeny virus from PEL cells at low concentrations, which do not affect PEL cell growth. These findings suggest that proteasome

  17. MDM2 regulates hypoxic hypoxia-inducible factor 1α stability in an E3 ligase, proteasome, and PTEN-phosphatidylinositol 3-kinase-AKT-dependent manner.

    Science.gov (United States)

    Joshi, Shweta; Singh, Alok R; Durden, Donald L

    2014-08-15

    Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor containing an inducibly expressed HIF1α subunit and a constitutively expressed HIF1β subunit. Under hypoxic conditions, the HIF1α subunit accumulates because of a decrease in the rate of proteolytic degradation, and the resulting HIF1α-HIF1β heterodimers undergo post-translational modifications that promote transactivation. Previous reports suggest that amplified signaling through PI3K enhances HIF1-dependent gene expression; however, its role is controversial, and the mechanism is unclear. Using genetically engineered PTEN-deficient cell lines, we demonstrate that PTEN specifically inhibited the accumulation of HIF1α in response to hypoxia. Furthermore, we report that in glioblastoma cell lines, inhibition of PI3K pathway, using pan as well as isoform-specific PI3K inhibitors SF1126, PF4691502, BEZ-235, GDC0941, and TGX221 blocked the induction of HIF1α protein and its targets vascular endothelial growth factor, HK1, and GLUT1 mRNA in response to hypoxia. Herein, we describe the first evidence that HIF1α can be degraded under hypoxic conditions via the 26 S proteasome and that MDM2 is the E3 ligase that induces the hypoxic degradation of HIF1α. Moreover, the action of MDM2 on HIF1α under hypoxia occurs in the cytoplasm and is controlled by the PTEN-PI3K-AKT signaling axis. These data strongly suggest a new role for PTEN in the regulation of HIF1α and importantly that PI3K-AKT activation is required for the hypoxic stabilization of HIF1α and that hypoxia alone is not sufficient to render HIF1α resistant to proteasomal cleavage and degradation. Moreover, these findings suggest new therapeutic considerations for PI3K and/or AKT inhibitors for cancer therapeutics. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  18. MDM2 Regulates Hypoxic Hypoxia-inducible Factor 1α Stability in an E3 Ligase, Proteasome, and PTEN-Phosphatidylinositol 3-Kinase-AKT-dependent Manner*

    Science.gov (United States)

    Joshi, Shweta; Singh, Alok R.; Durden, Donald L.

    2014-01-01

    Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor containing an inducibly expressed HIF1α subunit and a constitutively expressed HIF1β subunit. Under hypoxic conditions, the HIF1α subunit accumulates because of a decrease in the rate of proteolytic degradation, and the resulting HIF1α–HIF1β heterodimers undergo post-translational modifications that promote transactivation. Previous reports suggest that amplified signaling through PI3K enhances HIF1-dependent gene expression; however, its role is controversial, and the mechanism is unclear. Using genetically engineered PTEN-deficient cell lines, we demonstrate that PTEN specifically inhibited the accumulation of HIF1α in response to hypoxia. Furthermore, we report that in glioblastoma cell lines, inhibition of PI3K pathway, using pan as well as isoform-specific PI3K inhibitors SF1126, PF4691502, BEZ-235, GDC0941, and TGX221 blocked the induction of HIF1α protein and its targets vascular endothelial growth factor, HK1, and GLUT1 mRNA in response to hypoxia. Herein, we describe the first evidence that HIF1α can be degraded under hypoxic conditions via the 26 S proteasome and that MDM2 is the E3 ligase that induces the hypoxic degradation of HIF1α. Moreover, the action of MDM2 on HIF1α under hypoxia occurs in the cytoplasm and is controlled by the PTEN-PI3K-AKT signaling axis. These data strongly suggest a new role for PTEN in the regulation of HIF1α and importantly that PI3K-AKT activation is required for the hypoxic stabilization of HIF1α and that hypoxia alone is not sufficient to render HIF1α resistant to proteasomal cleavage and degradation. Moreover, these findings suggest new therapeutic considerations for PI3K and/or AKT inhibitors for cancer therapeutics. PMID:24982421

  19. Influence of combinations of fenugreek, garlic, and black pepper powder on production traits of the broilers

    Directory of Open Access Journals (Sweden)

    A. Kirubakaran

    2016-05-01

    Full Text Available Aim: To study the effects of combinations of fenugreek (Trigonella foenum-graecum L., garlic (Allium sativum, and black pepper (Piper nigrum powder supplementation on production traits of broiler chickens. Materials and Methods: A total of 288 commercial broiler chicks were randomly assigned to 1-9 groups with 4 replicates each. An experiment was conducted in broilers with different feed formulations; control feed, with no added fenugreek, garlic, and black pepper powder; and 8 treatment groups receiving feed supplemented with different combinations of fenugreek, garlic, and black pepper powder. The individual broilers’ body weight and feed consumption were recorded and calculate the body weight gain and feed conversion ratio (FCR. Results: Broiler’s weight gain and FCR were significantly higher in groups receiving feed supplemented with garlic and black pepper powder combinations (p<0.01. Cumulative feed consumption was significantly higher in groups receiving feed supplemented with garlic and black pepper powder combinations (p<0.01. Conclusion: The combination of garlic and black pepper powder supplemented broiler feed fed groups showed higher production performance. The 5 g/kg garlic powder+1 g/kg black pepper powder and 10 g/kg garlic powder+2 g/kg black pepper powder significantly improved the weight gain and FCR.

  20. A seasonal model of contracts between a monopsonistic processor and smallholder pepper producers in Costa Rica

    NARCIS (Netherlands)

    Sáenz Segura, F.; Haese, D' M.F.C.; Schipper, R.A.

    2010-01-01

    We model the contractual arrangements between smallholder pepper (Piper nigrum L.) producers and a single processor in Costa Rica. Producers in the El Roble settlement sell their pepper to only one processing firm, which exerts its monopsonistic bargaining power by setting the purchase price of

  1. Tospoviruses and Thrips and Integrated Resistance Management Strategies in Pepper in Florida

    Science.gov (United States)

    Florida ranks second in the production and value of bell pepper in the U.S. In 2015, Florida produced over one-half billion pounds of bell pepper on over 12,000 acres, valued at over 220 million dollars. In recent years, several invasive species of thrips and thrips-vectored tospoviruses have beco...

  2. Diversity and activity of biosurfactant-producing Pseudomonas in the rhizosphere of black pepper in Vietnam

    NARCIS (Netherlands)

    Tran, H.; Kruijt, M.; Raaijmakers, J.M.

    2008-01-01

    Aims: Phytophthora capsici is a major pathogen of black pepper and zoospores play an important role in the infection process. Fluorescent pseudomonads that produce biosurfactants with zoosporicidal activities were isolated from the black pepper rhizosphere in Vietnam, and their genotypic diversity

  3. Screening of pepper accessions for resistance against two thrips species (Frankliniella occidentalis and Thrips parvispinus)

    NARCIS (Netherlands)

    Maharijaya, A.; Vosman, B.; Steenhuis-Broers, M.M.; Harpenas, Asep; Purwito, A.; Visser, R.G.F.; Voorrips, R.E.

    2011-01-01

    Thrips are damaging pests in pepper worldwide. They can cause damage directly by feeding on leaves, fruits or flowers, and also indirectly by transferring viruses, especially tomato spotted wilt virus (TSWV). Although thrips are among the most damaging pests in pepper, until now there is no

  4. A semiochemical-based push-pull management strategy for pepper weevil

    Science.gov (United States)

    The pepper weevil Anthonomus eugeenii is a serious pest on peppers in southern United States. The weevils lay their eggs in flower buds and immature fruit where the larvae feed on the developing seed. Consequently, infestations are hard to control by pesticide applications. The aggregation pheromo...

  5. 7 CFR 319.56-24 - Lettuce and peppers from Israel.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Lettuce and peppers from Israel. 319.56-24 Section 319... Lettuce and peppers from Israel. (a) Lettuce may be imported into the United States from Israel without... protected with sticky traps and prophylactic sprays approved for the crop by Israel; (v) The lettuce must be...

  6. Effect of cultivar and roasting technique on sensory quality of Bierzo roasted pepper.

    Science.gov (United States)

    Guerra, Marcos; Sanz, Miguel A; Valenciano, José B; Casquero, Pedro A

    2011-10-01

    Pepper (Capsicum annuum L.) is one of the main horticultural products in the world. Roasted pepper is a high quality transformed product in the Iberian Peninsula, and obtained the recognition of 'Protected Geographical Indication' (PGI) of 'Pimiento Asado del Bierzo' in 2002. Roasted pepper has been traditionally processed with a steel-sheet hob. However, there are no data available about the effect of roasting technique in the quality of roasted pepper. The objective of this work was to compare the sensory quality of roasted pepper using industrial roasting techniques. Sensory properties that showed significant differences between roasting techniques were colour, thickness and charred remains (appearance descriptors), bitterness (taste descriptor) and smokiness (after-taste descriptor). Higher value of descriptors such as colour, charred remains and smokiness for peppers elaborated in a rotary oven, helped roasted pepper to reach a higher level of overall quality, although rotary oven samples reached the lowest roast yield. Roasting technique, rather than landrace, had the greatest effect on the sensory quality of roasted pepper, so the rotary oven was the roasting technique that achieved the highest quality score. This will contribute to improve sensory quality and marketing of PGI 'Pimiento Asado del Bierzo' in high quality markets. Copyright © 2011 Society of Chemical Industry.

  7. Fruit cuticle lipid composition and water loss in a diverse collection of pepper (capsicum)

    Science.gov (United States)

    Pepper (Capsicum spp.) fruits are covered by a relatively thick coating of cuticle that limits fruit water loss, a trait previously associated with maintenance of post-harvest fruit quality during commercial marketing. We’ve examined the fruit cuticles from 50 diverse pepper genotypes from a world c...

  8. Integrated crop management of hot pepper (Capsicum spp.) in tropical lowlands

    NARCIS (Netherlands)

    Vos, J.G.M.

    1994-01-01

    Hot pepper ( Capsicum spp.) is the most important low elevation vegetable commodity in Indonesia. Yields are low, in part due to crop health problems. Farmers' practices were surveyed by means of exploratory surveys. Hot pepper pests and diseases were identified and

  9. Diversity and activity of biosurfactant-producing Pseudomonas in the rhizosphere of black pepper in Vietnam.

    Science.gov (United States)

    Tran, H; Kruijt, M; Raaijmakers, J M

    2008-03-01

    Phytophthora capsici is a major pathogen of black pepper and zoospores play an important role in the infection process. Fluorescent pseudomonads that produce biosurfactants with zoosporicidal activities were isolated from the black pepper rhizosphere in Vietnam, and their genotypic diversity and potential to control Phy. capsici root rot was determined. Biosurfactant-producing pseudomonads were genotypically and biochemically characterized by BOX-polymerase chain reaction (PCR), 16S-rDNA sequencing, reverse-phase-high-performance liquid chromatography and liquid chromatography-mass spectrometry analyses. Biosurfactant-producing fluorescent pseudomonads make up c. 1.3% of the culturable Pseudomonas population in the rhizosphere of black pepper. Although BOX-PCR revealed substantial genotypic diversity, the isolates were shown to produce the same biosurfactants and were all identified as Pseudomonas putida. When applied to black pepper stem cuttings, several of the biosurfactant-producing strains provided significant disease control. In absence of the disease, several of the bacterial strains promoted shoot and root growth of black pepper stem cuttings. Biosurfactant-producing pseudomonads indigenous to the rhizosphere of black pepper plants are genotypically diverse and provide a novel resource for the control of Phy. capsici root rot and growth promotion of black pepper stem cuttings. The results of this study provide a strong basis for further development of supplementary strategies with antagonistic bacteria to control foot and root rot of black pepper and to promote plant growth.

  10. First Complete Genome Sequence of Pepper vein yellows virus from Australia

    Science.gov (United States)

    Maina, Solomon; Edwards, Owain R.

    2016-01-01

    We present here the first complete genomic RNA sequence of the polerovirus Pepper vein yellows virus (PeVYV) obtained from a pepper plant in Australia. We compare it with complete PeVYV genomes from Japan and China. The Australian genome was more closely related to the Japanese than the Chinese genome. PMID:27231375

  11. Baicalin and scutellarin are proteasome inhibitors that specifically target chymotrypsin-like catalytic activity.

    Science.gov (United States)

    Wu, Yi-Xin; Sato, Eiji; Kimura, Wataru; Miura, Naoyuki

    2013-09-01

    Baicalin and scutellarin are the major active principal flavonoids extracted from the Chinese herbal medicines Scutellaria baicalensis and Erigeron breviscapus (Vant.) Hand-Mazz. It has recently been reported that baicalin and scutellarin have antitumor activity. However, the mechanisms of action are unknown. We previously reported that some flavonoids have a specific role in the inhibition of the activity of proteasome subunits and induced apoptosis in tumor cells. To further investigate these pharmacological effects, we examined the inhibitory activity of baicalin and scutellarin on the extracted proteasomes from mice and cancer cells. Using fluorogenic substrates for proteasome catalytic subunits, we found that baicalin and scutellarin specifically inhibited chymotrypsin-like activity but did not inhibit trypsin-like and peptidyl-glutamyl peptide hydrolyzing activities. These data suggested that baicalin and scutellarin specifically inhibit chymotrypsin-like catalytic activity in the proteasome. Copyright © 2012 John Wiley & Sons, Ltd.

  12. Molecular mechanisms for synergistic effect of proteasome inhibitors with platinum-based therapy in solid tumors.

    Science.gov (United States)

    Chao, Angel; Wang, Tzu-Hao

    2016-02-01

    The successful development of the proteasome inhibitor bortezomib as an anticancer drug has improved survival in patients with multiple myeloma. With the emergence of the newly US Food and Drug Administration-approved proteasome inhibitor carfilzomib, ongoing trials are investigating this compound and other proteasome inhibitors either alone or in combination with other chemotherapy drugs. However, in solid tumors, the efficacy of proteasome inhibitors has not lived up to expectations. Results regarding the potential clinical efficacy of bortezomib combined with other agents in the treatment of solid tumors are eagerly awaited. Recent identification of the molecular mechanisms (involving apoptosis and autophagy) by which bortezomib and cisplatin can overcome chemotherapy resistance and sensitize tumor cells to anticancer therapy can provide insights into the development of novel therapeutic strategies for patients with solid malignancies. Copyright © 2016. Published by Elsevier B.V.

  13. The ubiquitin proteasome system in glia and its role in neurodegenerative diseases

    NARCIS (Netherlands)

    Jansen, Anne H. P.; Reits, Eric A. J.; Hol, Elly M.

    2014-01-01

    The ubiquitin proteasome system (UPS) is crucial for intracellular protein homeostasis and for degradation of aberrant and damaged proteins. The accumulation of ubiquitinated proteins is a hallmark of many neurodegenerative diseases, including amyotrophic lateral sclerosis, Alzheimer's, Parkinson's,

  14. Proteasome inhibitors enhance endothelial thrombomodulin expression via induction of Krüppel-like transcription factors

    NARCIS (Netherlands)

    Hiroi, Toyoko; Deming, Clayton B.; Zhao, Haige; Hansen, Baranda S.; Arkenbout, Elisabeth K.; Myers, Thomas J.; McDevitt, Michael A.; Rade, Jeffrey J.

    2009-01-01

    OBJECTIVE: Impairment of the thrombomodulin-protein C anticoagulant pathway has been implicated in pathological thrombosis associated with malignancy. Patients who receive proteasome inhibitors as part of their chemotherapeutic regimen appear to be at decreased risk for thromboembolic events. We

  15. The effect of irradiation in the preservation of pink pepper (Schinus terebinthifolius Raddi)

    Science.gov (United States)

    de Souza, Adriana Régia Marques; Arthur, Valter; Nogueira, Danielle Pires

    2012-08-01

    Pink peppers, also known as "pimenta-rosa" and "poivre rose", are the fruit of Schinus terebinthifolius Raddi, a species of pepper cultivated in Brazil, and have great potential for the exploration of uses. In efforts to lengthen the shelf life of this pepper, the purpose of this study was to evaluate the effect of different doses of radiation on its physical composition and color. The pink pepper samples were irradiated with doses of 0, 0.2, 0.4, 0.8 and 1.6 kGy, and the moisture, ash and lipid contents, pH and color were analyzed. The moisture content, lipid content and pH analysis indicated effects due to the irradiation (p>0.05) in which the higher doses resulted in decreases in the attribute. In contrast, there were no significant differences for the ash analysis (ppreservation process for pink peppers.

  16. Turmeric and black pepper spices decrease lipid peroxidation in meat patties during cooking

    Science.gov (United States)

    Zhang, Yanjun; Henning, Susanne M.; Lee, Ru-Po; Huang, Jianjun; Zerlin, Alona; Li, Zhaoping; Heber, David

    2015-01-01

    Abstract Spices are rich in natural antioxidants and have been shown to be potent inhibitors of lipid peroxidation during cooking of meat. Turmeric contains unique conjugated curcuminoids with strong antioxidant activity. Piperine, one of the main constituents of black pepper, is known to increase the bioavailability of curcuminoids in mouse and human studies when consumed with turmeric. We investigated whether adding black pepper to turmeric powder may further inhibit lipid peroxidation when added to meat patties prior to cooking. The addition of black pepper to turmeric significantly decreased the lipid peroxidation in hamburger meat. When investigating the antioxidant activity of the main chemical markers, we determined that piperine did not exhibit any antioxidant activity. Therefore, we conclude that other black pepper ingredients are responsible for the increased antioxidant activity of combining black pepper with turmeric powder. PMID:25582173

  17. Genome sequence of the hot pepper provides insights into the evolution of pungency in Capsicum species.

    Science.gov (United States)

    Kim, Seungill; Park, Minkyu; Yeom, Seon-In; Kim, Yong-Min; Lee, Je Min; Lee, Hyun-Ah; Seo, Eunyoung; Choi, Jaeyoung; Cheong, Kyeongchae; Kim, Ki-Tae; Jung, Kyongyong; Lee, Gir-Won; Oh, Sang-Keun; Bae, Chungyun; Kim, Saet-Byul; Lee, Hye-Young; Kim, Shin-Young; Kim, Myung-Shin; Kang, Byoung-Cheorl; Jo, Yeong Deuk; Yang, Hee-Bum; Jeong, Hee-Jin; Kang, Won-Hee; Kwon, Jin-Kyung; Shin, Chanseok; Lim, Jae Yun; Park, June Hyun; Huh, Jin Hoe; Kim, June-Sik; Kim, Byung-Dong; Cohen, Oded; Paran, Ilan; Suh, Mi Chung; Lee, Saet Buyl; Kim, Yeon-Ki; Shin, Younhee; Noh, Seung-Jae; Park, Junhyung; Seo, Young Sam; Kwon, Suk-Yoon; Kim, Hyun A; Park, Jeong Mee; Kim, Hyun-Jin; Choi, Sang-Bong; Bosland, Paul W; Reeves, Gregory; Jo, Sung-Hwan; Lee, Bong-Woo; Cho, Hyung-Taeg; Choi, Hee-Seung; Lee, Min-Soo; Yu, Yeisoo; Do Choi, Yang; Park, Beom-Seok; van Deynze, Allen; Ashrafi, Hamid; Hill, Theresa; Kim, Woo Taek; Pai, Hyun-Sook; Ahn, Hee Kyung; Yeam, Inhwa; Giovannoni, James J; Rose, Jocelyn K C; Sørensen, Iben; Lee, Sang-Jik; Kim, Ryan W; Choi, Ik-Young; Choi, Beom-Soon; Lim, Jong-Sung; Lee, Yong-Hwan; Choi, Doil

    2014-03-01

    Hot pepper (Capsicum annuum), one of the oldest domesticated crops in the Americas, is the most widely grown spice crop in the world. We report whole-genome sequencing and assembly of the hot pepper (Mexican landrace of Capsicum annuum cv. CM334) at 186.6× coverage. We also report resequencing of two cultivated peppers and de novo sequencing of the wild species Capsicum chinense. The genome size of the hot pepper was approximately fourfold larger than that of its close relative tomato, and the genome showed an accumulation of Gypsy and Caulimoviridae family elements. Integrative genomic and transcriptomic analyses suggested that change in gene expression and neofunctionalization of capsaicin synthase have shaped capsaicinoid biosynthesis. We found differential molecular patterns of ripening regulators and ethylene synthesis in hot pepper and tomato. The reference genome will serve as a platform for improving the nutritional and medicinal values of Capsicum species.

  18. Natural biostimulants reduce the incidence of BER in sweet yellow pepper plants (Capsicum annuum L.

    Directory of Open Access Journals (Sweden)

    Nada Parađiković

    2013-06-01

    Full Text Available Modern greenhouse pepper production should evolve towards more sustainable systems. The growing technique which combines soilless culture and biostimulants may reduce nutrient and water use with beneficial impact on the environment. Therefore, this work aimed to investigate effects of biostimulants application on hydroponically produced pepper plants (Capsicum annuum L. in conditions of reduced fertilization. Positive influence of biostimulant treatment on yield parameters was observed along with significant decrease in incidence of blossom-end rot (BER in two pepper cultivars. Biostimulants application resulted in overall increase in macro- and microelement content in fruits of treated pepper cultivars. Generally, biostimulants improved the yield of pepper plants grown hydroponically by increasing the nutrient uptake and decreasing the occurrence of BER. Thus, the application of biostimulants could be considered as a good production strategy for obtaining high yields of nutritionally valuable vegetables with lower impact on the environment.

  19. A monoclonal antibody that distinguishes latent and active forms of the proteasome (multicatalytic proteinase complex)

    Science.gov (United States)

    Weitman, D.; Etlinger, J. D.

    1992-01-01

    Monoclonal antibodies (mAbs) were generated to proteasome purified from human erythrocytes. Five of six proteasome-specific mAbs reacted with three subunits in the molecular mass range of 25-28 kDa, indicating a common epitope. The other mAb (AP5C10) exhibited a more restricted reactivity, recognizing a 32-kDa subunit of the proteasome purified in its latent state. However, when the proteasome is isolated in its active state, AP5C10 reacts with a 28-kDa subunit, evidence for processing of the proteasome subunits during purification. Purified proteasome preparations which exhibited partial latency have both AP5C10 reactive subunits. Although the 32-kDa subunit appears required for latency, loss of this component and generation of the 28-kDa component are not obligatory for activation. The 32- and 28-kDa subunits can each be further resolved into three components by isoelectric focusing. The apparent loss of 4 kDa during the conversion of the 32- to 28-kDa subunit is accompanied by a shift to a more basic pI for each polypeptide. Western blots of the early steps of proteasome purification reveal an AP5C10-reactive protein at 41 kDa. This protein was separated from proteasomes by sizing chromatography and may represent a pool of precursor subunits. Since the 32-kDa subunit appears necessary for latency, it is speculated to play a regulatory role in ATP-dependent proteolytic activity.

  20. Decreased proteasomal function accelerates cigarette smoke-induced pulmonary emphysema in mice.

    Science.gov (United States)

    Yamada, Yosuke; Tomaru, Utano; Ishizu, Akihiro; Ito, Tomoki; Kiuchi, Takayuki; Ono, Ayako; Miyajima, Syota; Nagai, Katsura; Higashi, Tsunehito; Matsuno, Yoshihiro; Dosaka-Akita, Hirotoshi; Nishimura, Masaharu; Miwa, Soichi; Kasahara, Masanori

    2015-06-01

    Chronic obstructive pulmonary disease (COPD) is a disease common in elderly people, characterized by progressive destruction of lung parenchyma and chronic inflammation of the airways. The pathogenesis of COPD remains unclear, but recent studies suggest that oxidative stress-induced apoptosis in alveolar cells contributes to emphysematous lung destruction. The proteasome is a multicatalytic enzyme complex that plays a critical role in proteostasis by rapidly destroying misfolded and modified proteins generated by oxidative and other stresses. Proteasome activity decreases with aging in many organs including lungs, and an age-related decline in proteasomal function has been implicated in various age-related pathologies. However, the role of the proteasome system in the pathogenesis of COPD has not been investigated. Recently, we have established a transgenic (Tg) mouse model with decreased proteasomal chymotrypsin-like activity, showing age-related phenotypes. Using this model, we demonstrate here that decreased proteasomal function accelerates cigarette smoke (CS)-induced pulmonary emphysema. CS-exposed Tg mice showed remarkable airspace enlargement and increased foci of inflammation compared with wild-type controls. Importantly, apoptotic cells were found in the alveolar walls of the affected lungs. Impaired proteasomal activity also enhanced apoptosis in cigarette smoke extract (CSE)-exposed fibroblastic cells derived from mice and humans in vitro. Notably, aggresome formation and prominent nuclear translocation of apoptosis-inducing factor were observed in CSE-exposed fibroblastic cells isolated from Tg mice. Collective evidence suggests that CS exposure and impaired proteasomal activity coordinately enhance apoptotic cell death in the alveolar walls that may be involved in the development and progression of emphysema in susceptible individuals such as the elderly.

  1. Validation of the 2nd Generation Proteasome Inhibitor Oprozomib for Local Therapy of Pulmonary Fibrosis.

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    Nora Semren

    Full Text Available Proteasome inhibition has been shown to prevent development of fibrosis in several organs including the lung. However, effects of proteasome inhibitors on lung fibrosis are controversial and cytotoxic side effects of the overall inhibition of proteasomal protein degradation cannot be excluded. Therefore, we hypothesized that local lung-specific application of a novel, selective proteasome inhibitor, oprozomib (OZ, provides antifibrotic effects without systemic toxicity in a mouse model of lung fibrosis. Oprozomib was first tested on the human alveolar epithelial cancer cell line A549 and in primary mouse alveolar epithelial type II cells regarding its cytotoxic effects on alveolar epithelial cells and compared to the FDA approved proteasome inhibitor bortezomib (BZ. OZ was less toxic than BZ and provided high selectivity for the chymotrypsin-like active site of the proteasome. In primary mouse lung fibroblasts, OZ showed significant anti-fibrotic effects, i.e. reduction of collagen I and α smooth muscle actin expression, in the absence of cytotoxicity. When applied locally into the lungs of healthy mice via instillation, OZ was well tolerated and effectively reduced proteasome activity in the lungs. In bleomycin challenged mice, however, locally applied OZ resulted in accelerated weight loss and increased mortality of treated mice. Further, OZ failed to reduce fibrosis in these mice. While upon systemic application OZ was well tolerated in healthy mice, it rather augmented instead of attenuated fibrotic remodelling of the lung in bleomycin challenged mice. To conclude, low toxicity and antifibrotic effects of OZ in pulmonary fibroblasts could not be confirmed for pulmonary fibrosis of bleomycin-treated mice. In light of these data, the use of proteasome inhibitors as therapeutic agents for the treatment of fibrotic lung diseases should thus be considered with caution.

  2. Modulation of the Proteasome Pathway by Nano-Curcumin and Curcumin in Retinal Pigment Epithelial Cells.

    Science.gov (United States)

    Ramos de Carvalho, J Emanuel; Verwoert, Milan T; Vogels, Ilse M C; Schipper-Krom, Sabine; Van Noorden, Cornelis J F; Reits, Eric A; Klaassen, Ingeborg; Schlingemann, Reinier O

    2018-01-01

    Curcumin has multiple biological effects including the modulation of protein homeostasis by the ubiquitin-proteasome system. The purpose of this study was to assess the in vitro cytotoxic and oxidative effects of nano-curcumin and standard curcumin and characterize their effects on proteasome regulation in retinal pigment epithelial (RPE) cells. Viability, cell cycle progression, and reactive oxygen species (ROS) production were determined after treatment with nano-curcumin or curcumin. Subsequently, the effects of nano-curcumin and curcumin on proteasome activity and the gene and protein expression of proteasome subunits PA28α, α7, β5, and β5i were assessed. Nano-curcumin (5-100 μM) did not show significant cytotoxicity or anti-oxidative effects against H2O2-induced oxidative stress, whereas curcumin (≥10 μM) was cytotoxic and a potent inducer of ROS production. Both nano-curcumin and curcumin induced changes in proteasome-mediated proteolytic activity characterized by increased activity of the proteasome subunits β2 and β5i/β1 and reduced activity of β5/β1i. Likewise, nano-curcumin and curcumin affected mRNA and protein levels of household and immunoproteasome subunits. Nano-curcumin is less toxic to RPE cells and less prone to induce ROS production than curcumin. Both nano-curcumin and curcumin increase proteasome-mediated proteolytic activity. These results suggest that nano-curcumin may be regarded as a proteasome-modulating agent of limited cytotoxicity for RPE cells. The Author(s). Published by S. Karger AG, Basel.

  3. Inhibitors of the proteasome reduce the accelerated proteolysis in atrophying rat skeletal muscles.

    OpenAIRE

    Tawa, N E; Odessey, R; Goldberg, A L

    1997-01-01

    Several observations have suggested that the enhanced proteolysis and atrophy of skeletal muscle in various pathological states is due primarily to activation of the ubiquitin-proteasome pathway. To test this idea, we investigated whether peptide aldehyde inhibitors of the proteasome, N-acetyl-leucyl-leucyl-norleucinal (LLN), or the more potent CBZ-leucyl-leucyl-leucinal (MG132) suppressed proteolysis in incubated rat skeletal muscles. These agents (e.g., MG132 at 10 microM) inhibited nonlyso...

  4. BAX/BAK–Independent Mitoptosis during Cell Death Induced by Proteasome Inhibition?

    OpenAIRE

    Lomonosova, Elena; Ryerse, Jan; Chinnadurai, G.

    2009-01-01

    Proteasome inhibitors induce rapid death of cancer cells. We show that in epithelial cancer cells, such death is associated with dramatic and simultaneous up-regulation of several BH3-only proteins, including BIK, BIM, MCL-1S, NOXA, and PUMA, as well as p53. Elevated levels of these proteins seem to be the result of direct inhibition of their proteasomal degradation, induction of transcription, and active translation. Subsequent cell death is independent of BAX, and probably BAK, and proceeds...

  5. The Anti-Apoptotic Activity of BAG3 Is Restricted by Caspases and the Proteasome

    OpenAIRE

    Virador, Victoria M.; Davidson, Ben; Czechowicz, Josephine; Mai, Alisha; Kassis, Jareer; Kohn, Elise C.

    2009-01-01

    Background Caspase-mediated cleavage and proteasomal degradation of ubiquitinated proteins are two independent mechanisms for the regulation of protein stability and cellular function. We previously reported BAG3 overexpression protected ubiquitinated clients, such as AKT, from proteasomal degradation and conferred cytoprotection against heat shock. We hypothesized that the BAG3 protein is regulated by proteolysis. Methodology/Principal Findings Staurosporine (STS) was used as a tool to test ...

  6. Pepper seed variety identification based on visible/near-infrared spectral technology

    Science.gov (United States)

    Li, Cuiling; Wang, Xiu; Meng, Zhijun; Fan, Pengfei; Cai, Jichen

    2016-11-01

    Pepper is a kind of important fruit vegetable, with the expansion of pepper hybrid planting area, detection of pepper seed purity is especially important. This research used visible/near infrared (VIS/NIR) spectral technology to detect the variety of single pepper seed, and chose hybrid pepper seeds "Zhuo Jiao NO.3", "Zhuo Jiao NO.4" and "Zhuo Jiao NO.5" as research sample. VIS/NIR spectral data of 80 "Zhuo Jiao NO.3", 80 "Zhuo Jiao NO.4" and 80 "Zhuo Jiao NO.5" pepper seeds were collected, and the original spectral data was pretreated with standard normal variable (SNV) transform, first derivative (FD), and Savitzky-Golay (SG) convolution smoothing methods. Principal component analysis (PCA) method was adopted to reduce the dimension of the spectral data and extract principal components, according to the distribution of the first principal component (PC1) along with the second principal component(PC2) in the twodimensional plane, similarly, the distribution of PC1 coupled with the third principal component(PC3), and the distribution of PC2 combined with PC3, distribution areas of three varieties of pepper seeds were divided in each twodimensional plane, and the discriminant accuracy of PCA was tested through observing the distribution area of samples' principal components in validation set. This study combined PCA and linear discriminant analysis (LDA) to identify single pepper seed varieties, results showed that with the FD preprocessing method, the discriminant accuracy of pepper seed varieties was 98% for validation set, it concludes that using VIS/NIR spectral technology is feasible for identification of single pepper seed varieties.

  7. Microbial quality and bioactive constituents of sweet peppers from sustainable production systems.

    Science.gov (United States)

    Marín, Alicia; Gil, María I; Flores, Pilar; Hellín, Pilar; Selma, María V

    2008-12-10

    Integrated, organic, and soil-less production systems are the principal production practices that have emerged to encourage more sustainable agricultural practices and safer edible plants, reducing inputs of plaguicides, pesticides, and fertilizers. Sweet peppers grown commercially under integrated, organic, and soil-less production systems were compared to study the influence of these sustainable production systems on the microbial quality and bioactive constituents (vitamin C, individual and total carotenoids, hydroxycinnamic acids, and flavonoids). The antioxidant composition of peppers was analyzed at green and red maturity stages and at three harvest times (initial, middle, and late season). Irrigation water, manure, and soil were shown to be potential transmission sources of pathogens to the produce. Coliform counts of soil-less peppers were up to 2.9 log units lower than those of organic and integrated peppers. Soil-less green and red peppers showed maximum vitamin C contents of 52 and 80 mg 100 g(-1) fresh weight (fw), respectively, similar to those grown in the organic production system. Moreover, the highest content of total carotenoids was found in the soil-less red peppers, which reached a maximum of 148 mg 100 g(-1) fw, while slightly lower contents were found in integrated and organic red peppers. Hydroxycinnamic acids and flavonoids represented 15 and 85% of the total phenolic content, respectively. Total phenolic content, which ranged from 1.2 to 4.1 mg 100 g(-1) fw, was significantly affected by the harvest time but not by the production system assayed. Soil-less peppers showed similar or even higher concentrations of bioactive compounds (vitamin C, provitamin A, total carotenoid, hydroxycinnamic acids, and flavonoids) than peppers grown under organic and integrated practices. Therefore, in the commercial conditions studied, soil-less culture was a more suitable alternative than organic or integrated practices, because it improved the microbial

  8. Characterisation of 20S Proteasome in Tritrichomonas foetus and Its Role during the Cell Cycle and Transformation into Endoflagellar Form.

    Directory of Open Access Journals (Sweden)

    Antonio Pereira-Neves

    Full Text Available Proteasomes are intracellular complexes that control selective protein degradation in organisms ranging from Archaea to higher eukaryotes. These structures have multiple proteolytic activities that are required for cell differentiation, replication and maintaining cellular homeostasis. Here, we document the presence of the 20S proteasome in the protist parasite Tritrichomonas foetus. Complementary techniques, such as a combination of whole genome sequencing technologies, bioinformatics algorithms, cell fractionation and biochemistry and microscopy approaches were used to characterise the 20S proteasome of T. foetus. The 14 homologues of the typical eukaryotic proteasome subunits were identified in the T. foetus genome. Alignment analyses showed that the main regulatory and catalytic domains of the proteasome were conserved in the predicted amino acid sequences from T. foetus-proteasome subunits. Immunofluorescence assays using an anti-proteasome antibody revealed a labelling distributed throughout the cytosol as punctate cytoplasmic structures and in the perinuclear region. Electron microscopy of a T. foetus-proteasome-enriched fraction confirmed the presence of particles that resembled the typical eukaryotic 20S proteasome. Fluorogenic assays using specific peptidyl substrates detected presence of the three typical peptidase activities of eukaryotic proteasomes in T. foetus. As expected, these peptidase activities were inhibited by lactacystin, a well-known specific proteasome inhibitor, and were not affected by inhibitors of serine or cysteine proteases. During the transformation of T. foetus to endoflagellar form (EFF, also known as pseudocyst, we observed correlations between the EFF formation rates, increases in the proteasome activities and reduced levels of ubiquitin-protein conjugates. The growth, cell cycle and EFF transformation of T. foetus were inhibited after treatment with lactacystin in a dose-dependent manner. Lactacystin

  9. Prehispanic use of chili peppers in Chiapas, Mexico.

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    Terry G Powis

    Full Text Available The genus Capsicum is New World in origin and represents a complex of a wide variety of both wild and domesticated taxa. Peppers or fruits of Capsicum species rarely have been identified in the paleoethnobotanical record in either Meso- or South America. We report here confirmation of Capsicum sp. residues from pottery samples excavated at Chiapa de Corzo in southern Mexico dated from Middle to Late Preclassic periods (400 BCE to 300 CE. Residues from 13 different pottery types were collected and extracted using standard techniques. Presence of Capsicum was confirmed by ultra-performance liquid chromatography (UPLC/MS-MS Analysis. Five pottery types exhibited chemical peaks for Capsicum when compared to the standard (dihydrocapsaicin. No peaks were observed in the remaining eight samples. Results of the chemical extractions provide conclusive evidence for Capsicum use at Chiapas de Corzo during a 700 year period (400 BCE-300 CE. Presence of Capsicum in different types of culinary-associated pottery raises questions how chili pepper could have been used during this early time period. As Pre-Columbian cacao products sometimes were flavored using Capsicum, the same pottery sample set was tested for evidence of cacao using a theobromine marker: these results were negative. As each vessel that tested positive for Capsicum had a culinary use we suggest here the possibility that chili residues from the Chiapas de Corzo pottery samples reflect either paste or beverage preparations for religious, festival, or every day culinary use. Alternatively, some vessels that tested positive merely could have been used to store peppers. Most interesting from an archaeological context was the presence of Capsicum residue obtained from a spouted jar, a pottery type previously thought only to be used for pouring liquids.

  10. Proteasome (Prosome Subunit Variations during the Differentiation of Myeloid U937 Cells

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    Laurent Henry

    1997-01-01

    Full Text Available 20S proteasomes (prosomes/multicatalytic proteinase are protein particles built of 28 subunits in variable composition. We studied the changes in proteasome subunit composition during the differentiation of U937 cells induced by phorbol‐myristate‐acetate or retinoic acid plus 1,25‐dihydroxy‐cholecalciferol by western blot, flow cytometry and immuno‐fluorescence. p25K (C3, p27K (IOTA and p30/33K (C2 subunits were detected in both the nucleus and cytoplasm of undifferentiated cells. Flow cytometry demonstrated a biphasic decrease in proteasome subunits detection during differentiation induced by RA+VD. PMA caused an early transient decrease in these subunits followed by a return to their control level, except for p30/33K, which remained low. Immuno‐fluorescence also showed differences in the cytolocalization of the subunits, with a particular decrease in antigen labeling in the nucleus of RA+VD‐induced cells, and a scattering in the cytoplasm and a reorganization in the nucleus of PMA‐induced cells. Small amounts of proteasomal proteins were seen on the outer membrane of non‐induced cells; these membrane proteins disappeared when treated with RA+VD, whereas some increased on PMA‐induced cells. The differential changes in the distribution and type of proteasomes in RA+VD and PMA‐induced cells indicate that, possibly, 20S proteasomes may play a role in relation to the mechanisms of differentiation and the inducer used.

  11. Assessing subunit dependency of the Plasmodium proteasome using small molecule inhibitors and active site probes.

    Science.gov (United States)

    Li, Hao; van der Linden, Wouter A; Verdoes, Martijn; Florea, Bogdan I; McAllister, Fiona E; Govindaswamy, Kavitha; Elias, Joshua E; Bhanot, Purnima; Overkleeft, Herman S; Bogyo, Matthew

    2014-08-15

    The ubiquitin-proteasome system (UPS) is a potential pathway for therapeutic intervention for pathogens such as Plasmodium, the causative agent of malaria. However, due to the essential nature of this proteolytic pathway, proteasome inhibitors must avoid inhibition of the host enzyme complex to prevent toxic side effects. The Plasmodium proteasome is poorly characterized, making rational design of inhibitors that induce selective parasite killing difficult. In this study, we developed a chemical probe that labels all catalytic sites of the Plasmodium proteasome. Using this probe, we identified several subunit selective small molecule inhibitors of the parasite enzyme complex. Treatment with an inhibitor that is specific for the β5 subunit during blood stage schizogony led to a dramatic decrease in parasite replication while short-term inhibition of the β2 subunit did not affect viability. Interestingly, coinhibition of both the β2 and β5 catalytic subunits resulted in enhanced parasite killing at all stages of the blood stage life cycle and reduced parasite levels in vivo to barely detectable levels. Parasite killing was achieved with overall low host toxicity, something that has not been possible with existing proteasome inhibitors. Our results highlight differences in the subunit dependency of the parasite and human proteasome, thus providing a strategy for development of potent antimalarial drugs with overall low host toxicity.

  12. Antiatherogenic effect of quercetin is mediated by proteasome inhibition in the aorta and circulating leukocytes.

    Science.gov (United States)

    Pashevin, Denis A; Tumanovska, Lesya V; Dosenko, Victor E; Nagibin, Vasyl S; Gurianova, Veronika L; Moibenko, Alexey A

    2011-01-01

    Quercetin, a plant-derived flavonoid, has attracted considerable attention as promising compound for heart disease prevention and therapy. It has been linked to decreased mortality from heart disease and decreased incidence of stroke. Here, we report new data showing the angioprotective properties of quercetin mediated by its effect on proteasomal proteolysis. This study was designed to investigate the ability of quercetin to modulate proteasomal activity in a rabbit model of cholesterol-induced atherosclerosis. First, we show proteasomal trypsin-like (TL) activity increased up to 2.4-fold, chymotrypsin-like (CTL) activity increased by up to 43% and peptidyl-glutamyl peptide-hydrolyzing (PGPH) activity increased by up to 10% after 8 weeks of a cholesterol-rich diet. A single intravenous injection of the water-soluble form of quercetin (Corvitin) significantly decreased proteasomal TL activity 1.85-fold in monocytes, and decreased the CTL and PGPH activities more than 2-fold in polymorphonuclear leukocytes (PMNL) after 2 h. Prolonged administration (1 month) of Corvitin to animals following a cholesterol-rich diet significantly decreased all types of proteolytic proteasome activities both in tissues and in circulating leukocytes and was associated with the reduction of atherosclerotic lesion areas in the aorta. Additionally, the pharmacological form of quercetin (Quertin) was shown to have an antiatherogenic effect and an ability to inhibit proteasome activities.

  13. Defective Proteasome Delivery of Polyubiquitinated Proteins by Ubiquilin-2 Proteins Containing ALS Mutations.

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    Lydia Chang

    Full Text Available Ubiquilin proteins facilitate delivery of ubiquitinated proteins to the proteasome for degradation. Interest in the proteins has been heightened by the discovery that gene mutations in UBQLN2 cause dominant inheritance of amyotrophic lateral sclerosis (ALS. However, the mechanisms by which the mutations cause ALS are not known. Here we report on the underlying defect of ubiquilin-2 proteins containing ALS-linked mutations in affecting proteasome-mediated degradation. We found that overexpression of ubiquilin-2 proteins containing any one of five different ALS mutations slow degradation of Myc, a prototypic proteasome substrate. Examination of coprecipitating proteins indicated that the mutant proteins are generally capable of binding polyubiquitinated proteins, but defective in binding the proteasome. GST-pulldown studies revealed that many of the mutants bind weaker to the S5a subunit of the proteasome, compared with wild type (WT ubiquilin-2 protein. The results suggest the mutant proteins are unable to deliver their captured cargo to the proteasome for degradation, which presumably leads to toxicity. Quantification of cell death is consistent with this idea. Measurement of protein turnover further indicated the mutant proteins have longer half-lives than WT ubiquilin-2. Our studies provide novel insight into the mechanism by which ALS-linked mutations in UBQLN2 interfere with protein degradation.

  14. The ubiquitin-proteasome system and chromosome 17 in cerebellar granule cells and medulloblastoma subgroups.

    Science.gov (United States)

    Vriend, Jerry; Marzban, Hassan

    2017-02-01

    Chromosome 17 abnormalities are often observed in medulloblastomas (MBs), particularly those classified in the consensus Groups 3 and 4. Herein we review MB signature genes associated with chromosome 17 and the relationship of these signature genes to the ubiquitin-proteasome system. While clinical investigators have not focused on the ubiquitin-proteasome system in relation to MB, a substantial amount of data on the topic has been hidden in the form of supplemental datasets of gene expression. A supplemental dataset associated with the Thompson classification of MBs shows that a subgroup of MB with 17p deletions is characterized by reduced expression of genes for several core particle subunits of the beta ring of the proteasome (β1, β4, β5, β7). One of these genes (PSMB6, the gene for the β1 subunit) is located on chromosome 17, near the telomeric end of 17p. By comparison, in the WNT group of MBs only one core proteasome subunit, β6, associated with loss of a gene (PSMB1) on chromosome 6, was down-regulated in this dataset. The MB subgroups with the worst prognosis have a significant association with chromosome 17 abnormalities and irregularities of APC/C cyclosome genes. We conclude that the expression of proteasome subunit genes and genes for ubiquitin ligases can contribute to prognostic classification of MBs. The therapeutic value of targeting proteasome subunits and ubiquitin ligases in the various subgroups of MB remains to be determined separately for each classification of MB.

  15. Role of the fermentation process in off-odorant formation in white pepper: on-site trial in Thailand.

    Science.gov (United States)

    Steinhaus, Martin; Schieberle, Peter

    2005-07-27

    In white pepper samples, directly taken from a retting batch at a pepper production plant in Thailand, 3-methylindole, 4-methylphenol, 3-methylphenol, and butanoic acid, recently confirmed to be responsible for the characteristic fecal off-odor frequently detected in white pepper powders, were quantified by stable isotope dilution analyses. The results clearly showed that, in particular, 3-methylindole (fecal, swine-manure-like), 4-methylphenol (fecal, horse-like), and butanoic acid (cheese-like) were biochemically formed during retting, thus indicating that fermentation is the crucial step for off-odorant formation during white pepper processing. Model fermentation experiments performed with different manufacturing regimens revealed that white pepper, containing no substantial amounts of these odorants, can be produced from ripe starting material by a short fermentation under water and with frequent exchange of water. The overall aroma of such pepper was superior as compared to the aroma of white pepper produced according to the traditional procedure.

  16. Spaceflight Activates Autophagy Programs and the Proteasome in Mouse Liver.

    Science.gov (United States)

    Blaber, Elizabeth A; Pecaut, Michael J; Jonscher, Karen R

    2017-09-27

    Increased oxidative stress is an unavoidable consequence of exposure to the space environment. Our previous studies showed that mice exposed to space for 13.5 days had decreased glutathione levels, suggesting impairments in oxidative defense. Here we performed unbiased, unsupervised and integrated multi-'omic analyses of metabolomic and transcriptomic datasets from mice flown aboard the Space Shuttle Atlantis. Enrichment analyses of metabolite and gene sets showed significant changes in osmolyte concentrations and pathways related to glycerophospholipid and sphingolipid metabolism, likely consequences of relative dehydration of the spaceflight mice. However, we also found increased enrichment of aminoacyl-tRNA biosynthesis and purine metabolic pathways, concomitant with enrichment of genes associated with autophagy and the ubiquitin-proteasome. When taken together with a downregulation in nuclear factor (erythroid-derived 2)-like 2-mediated signaling, our analyses suggest that decreased hepatic oxidative defense may lead to aberrant tRNA post-translational processing, induction of degradation programs and senescence-associated mitochondrial dysfunction in response to the spaceflight environment.

  17. Spaceflight Activates Autophagy Programs and the Proteasome in Mouse Liver

    Directory of Open Access Journals (Sweden)

    Elizabeth A. Blaber

    2017-09-01

    Full Text Available Increased oxidative stress is an unavoidable consequence of exposure to the space environment. Our previous studies showed that mice exposed to space for 13.5 days had decreased glutathione levels, suggesting impairments in oxidative defense. Here we performed unbiased, unsupervised and integrated multi-‘omic analyses of metabolomic and transcriptomic datasets from mice flown aboard the Space Shuttle Atlantis. Enrichment analyses of metabolite and gene sets showed significant changes in osmolyte concentrations and pathways related to glycerophospholipid and sphingolipid metabolism, likely consequences of relative dehydration of the spaceflight mice. However, we also found increased enrichment of aminoacyl-tRNA biosynthesis and purine metabolic pathways, concomitant with enrichment of genes associated with autophagy and the ubiquitin-proteasome. When taken together with a downregulation in nuclear factor (erythroid-derived 2-like 2-mediated signaling, our analyses suggest that decreased hepatic oxidative defense may lead to aberrant tRNA post-translational processing, induction of degradation programs and senescence-associated mitochondrial dysfunction in response to the spaceflight environment.

  18. Effect of Red Pepper (Capsicum frutescens) Powder or Red Pepper Pigment on the Performance and Egg Yolk Color of Laying Hens.

    Science.gov (United States)

    Li, Huaqiang; Jin, Liji; Wu, Feifei; Thacker, Philip; Li, Xiaoyu; You, Jiansong; Wang, Xiaoyan; Liu, Sizhao; Li, Shuying; Xu, Yongping

    2012-11-01

    Two experiments were conducted to study the effects of red pepper (Capsicum frutescens) powder or red pepper pigment on the performance and egg yolk color of laying hens. In Exp. 1, 210, thirty-wk old, Hy-line Brown laying hens were fed one of seven diets containing 0.3, 0.6, 1.2, 2.0, 4.8 or 9.6 ppm red pepper pigment or 0.3 ppm carophyll red. Each diet was fed to three replicate batteries of hens with each battery consisting of a row of five cages of hens with two hens per cage (n = 3). In Exp. 2, 180, thirty-wk old, Hyline Brown laying hens, housed similarly to those in Exp. 1, were fed an unsupplemented basal diet as well as treatments in which the basal diet was supplemented with 0.8% red pepper powder processed in a laboratory blender to an average particle size of 300 μm, 0.8% red pepper powder processed as a super fine powder with a vibrational mill (44 μm) and finally 0.8% red pepper powder processed as a super fine powder with a vibrational mill but mixed with 5% Na2CO3 either before or after grinding. A diet supplemented with 0.3 ppm carophyll red pigment was also included (n = 3). In both experiments, hens were fed the red pepper powder or pigment for 14 days. After feeding of the powder or pigment was terminated, all hens were fed the basal diet for eight more days to determine if the dietary treatments had any residual effects. In Exp. 1, there were no differences in egg-laying performance, feed consumption or feed conversion ratio due to inclusion of red pepper pigment in the diet. Average egg weight was higher (pfeeding red pepper powder did not affect egg-laying performance, feed consumption or feed conversion ratio (p>0.05). However, compared with the control group, supplementation with all of the red pepper powder treatments increased egg weight (pfeed additives for improving egg yolk color for laying hens.

  19. Evaluation of antioxidant potential of pepper sauce (Capsicum frutescens L.

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    Pamela Freire de Moura Pereira

    2016-12-01

    Full Text Available Functional properties of substances present in in natura foods such as fruits and vegetables are well documented; however, the activity that remains after processing needs more research. The present study aimed to evaluate the antioxidant potential in fruit processed as sauce and quantify the compounds able to contribute to such activity. Three different treatments were developed varying only the concentration of pepper Capsicum frutescens L., with treatment ratios (fruit: water: vinegar: salt being: treatment 1 (0.5: 1: 0.5: 0.33, 2 (1: 1: 0.5: 0.33, and 3 (2: 1: 0.5: 0.33. By the DPPH method, the values found for EC50 (g g DPPH−1 from 3726.9 to 5425.9 for the alcoholic extract were the most significant. The content of total phenols did not vary between the three treatments. While the content of carotenoids found was significantly different in the treatment with lower content of the fruit in natura, when compared to the treatment with higher content (44.02 and 56.09 μg of β-carotene 100 g−1, respectively and the content of ascorbic acid varied between 10.95 and 21.59 mg 100−1 g. Therefore, the pepper sauce was presented as an alternative to the consumption of bioactive compounds that may have antioxidant potential.

  20. Radiation sterilization of red chili pepper (Capsicum frutescens)

    International Nuclear Information System (INIS)

    Onyenekwe, P.C.; Ogbadu, G.H.

    1995-01-01

    The effect of radapertization using 60Co gamma radiation on the chemical and organoleptic properties of ground and whole dry pungent red pepper has been investigated. The fungal population was eliminated with a dose of 7.5 kGy and a dose of 10 kGy was required to radapertize the samples. Irradiation up to 10 kGy did not produce any significant (P 0.05) changes in the contents of capsaicin and carbohydrate. Oleoresin content was significantly (P 0.05) increased from 24.45 to 31.61% and lipid from 16.80 to 19.30%. The observed effect on apparent oleoresin and lipid contents was due to enhanced extractability. Storage time had no effect on the sensory properties of the spice but rather enhanced microbial load reduction in the medium dose (5.0 and 7.5 kGy) treated samples. A dose of 10 kGy has been certified to pose no health hazard to humans; hence decontamination of red pepper with up to 10 kGy gamma radiation does not require any further toxicological study and is therefore recommended

  1. Microbiological and Physicochemical Quality Characterization of Commercial Red Pepper Powders

    International Nuclear Information System (INIS)

    Jeong, M.S.; Ahn, J.J.; Kashig, A.; Kim, G.R.; Kwon, J.H.; Im, J.G.

    2013-01-01

    Ten commercially available red pepper powders were investigated for their hygienic quality (total aerobic count, yeasts and molds, and coliforms) and physicochemical properties (moisture content, pH, Hunters color values, American Spice Trade Association (ASTA) color and particle size). Microbial analysis resulted in 10 3 -10 6 CFU/g of total aerobic count and same of yeasts and molds, where 2 samples were positive (103 CFU/g) for coliforms. The moisture contents (7.25% to 12.73%) were with in the range as described in the Korean Food Standards Codex. Noteworthy variations were observed in pH (4.97 to 5.15), Hunters E values (47.19 to 58.04) and ASTA color values (89.31 to 98.61). Although the color differences were evident among the samples, but the Hunter values were not in good correlations with ASTA color values. The average particle sizes of the all samples were comparable ranging from 605 μm to 1251 μm with few exceptions. There was a great variation in the key quality attributes of commercially available red pepper powders that should be considered for their various uses in food products. (author)

  2. Ascorbic Acid Contents in Chili Peppers (Capsicum L.

    Directory of Open Access Journals (Sweden)

    Owk ANIEL KUMAR

    2009-12-01

    Full Text Available The genus Capsicum commonly known as chili pepper is a major spice crop and is almost cosmopolitan in distribution. The nutritive value of chili pepper is largely determined by ascorbic acid content. The fruits at five ripening stages viz., (M1, M2, M3, M4 and M5 from seventeen cultivars of Capsicum annuum L and one cultivar of Capsicum frutescens L were analyzed for ascorbic acid content. Among eighteen genotypes the C. annuum var. IC: 119262(CA2 showed higher ascorbic acid content (mg/100g FW i.e., 208.00.68 (M1, 231.00.66 (M2, 280.00.31 (M3, 253.00.34 (M4 and 173.70.27 (M5. The study revealed that the gradual increase in ascorbic acid content from green to red and subsequently declined in the lateral stages (red partially dried and red fully dried fruits. The variability of ascorbic acid content in the genotypes suggests that these selected genotypes may be use full as parents in hybridization programs to produce fruits with good nutritional values.

  3. Influence of radappertization upon some sensory properties of black pepper

    Energy Technology Data Exchange (ETDEWEB)

    Uchman, W; Fiszer, W; Pawlik, A [Wyzsza Szkola Rolnicza, Poznan (Poland)

    1983-01-01

    Radappertization may be used as an effective method for sterilization of spices. A possible influence of irradiation upon the sensoric qualities should be considered in this context. Therefore the objectives of the work were the determination of the influence of pepper irradiation on its sensoric quality, on the qualitative changes of volatile chemical substances and on the correlation between changes resulting from irradiation and the sensoric quality of pepper. The spice was irradiated with doses of 10, 0, 40 and 60 kGy. Thresholds of recognition and difference were determined. The quantity of the dose was related to the values of thresholds mentioned above. The larger the dose, the less intensive the flavor and the higher the thresholds. A significant influence of irradiation upon the decrease of the quantity of essential oils and carbohydrates was noticed. There was a relation between the results of sensoric assessments and the chemical characteristics (the content of volatile carbonyl, sulphuric and nitrogenic substances). Correlation between these factors was determined on the basic statistical analysis of the results obtained during investigation.

  4. Napropamide residues in runoff and infiltration water from pepper production.

    Science.gov (United States)

    Antonious, George F; Patterson, Matthew A

    2005-01-01

    A field study was conducted on a Lowell silty loam soil of 2.7% organic matter at the Kentucky State University Research Farm, Franklin County, Kentucky. Eighteen universal soil loss equation (USLE) standard plots (22 x 3.7 m each) were established on a 10% slope. Three soil management practices were used: (i) class-A biosolids (sewage sludge), (ii) yard waste compost, each mixed with native soil at a rate of 50 ton acre(-1) on a dry-weight basis, and (iii) a no-mulch (NM) treatment (rototilled bare soil), used for comparison purposes. Devrinol 50-DF "napropamide" [N,N-diethyl-2-(1-naphthyloxy) propionamide] was applied as a preemergent herbicide, incorporated into the soil surface, and the plots were planted with 60-day-old sweet bell pepper seedlings. Napropamide residues one hour following spraying averaged 0.8, 0.4, and 0.3 microg g(-1) dry soil in sewage sludge, yard waste compost, and no-mulch treatments, respectively. Surface runoff water, runoff sediment, and napropamide residues in runoff were significantly reduced by the compost and biosolid treatments. Yard waste compost treatments increased water infiltration and napropamide residues in the vadose zone compared to sewage sludge and NM treatments. Total pepper yields from yard waste compost amended soils (9187 lbs acre(-1)) was significantly higher (P soil amended with class-A biosolids (6984 lbs acre(-1)) or the no-mulch soil (7162 lbs acre(-1)).

  5. Electron spin resonance characterization of radical components in irradiated black pepper skin and core

    International Nuclear Information System (INIS)

    Yamaoki, Rumi; Kimura, Shojiro; Ohta, Masatoshi

    2011-01-01

    Characteristics of free radical components of irradiated black pepper fruit (skin) and the pepper seed (core) were analyzed using electron spin resonance. A weak signal near g=2.005 was observed in black pepper before irradiation. Complex spectra near g=2.005 with three lines (the skin) or seven lines (the core) were observed in irradiated black pepper (both end line width; ca. 6.8 mT). The spectral intensities decreased considerably at 30 days after irradiation, and continued to decrease steadily thereafter. The spectra simulated on the basis of the content and the stability of radical components derived from plant constituents, including fiber, starch, polyphenol, mono- and disaccharide, were in good agreement with the observed spectra. Analysis showed that the signal intensities derived from fiber in the skin for an absorbed dose were higher, and the rates of decrease were lower, than that in the core. In particular, the cellulose radical component in the skin was highly stable. - Highlights: → We identified the radical components in irradiated black pepper skin and core. → The ESR spectra near g=2.005 with 3-7 lines were emerged after irradiation. → Spectra simulated basing on the content and the stability of radical from the plant constituents. → Cellulose radical component in black pepper skin was highly stable. → Single signal near g=2.005 was the most stable in black pepper core.

  6. Whole-genome sequencing of cultivated and wild peppers provides insights into Capsicum domestication and specialization

    Science.gov (United States)

    Qin, Cheng; Yu, Changshui; Shen, Yaou; Fang, Xiaodong; Chen, Lang; Min, Jiumeng; Cheng, Jiaowen; Zhao, Shancen; Xu, Meng; Luo, Yong; Yang, Yulan; Wu, Zhiming; Mao, Likai; Wu, Haiyang; Ling-Hu, Changying; Zhou, Huangkai; Lin, Haijian; González-Morales, Sandra; Trejo-Saavedra, Diana L.; Tian, Hao; Tang, Xin; Zhao, Maojun; Huang, Zhiyong; Zhou, Anwei; Yao, Xiaoming; Cui, Junjie; Li, Wenqi; Chen, Zhe; Feng, Yongqiang; Niu, Yongchao; Bi, Shimin; Yang, Xiuwei; Li, Weipeng; Cai, Huimin; Luo, Xirong; Montes-Hernández, Salvador; Leyva-González, Marco A.; Xiong, Zhiqiang; He, Xiujing; Bai, Lijun; Tan, Shu; Tang, Xiangqun; Liu, Dan; Liu, Jinwen; Zhang, Shangxing; Chen, Maoshan; Zhang, Lu; Zhang, Li; Zhang, Yinchao; Liao, Weiqin; Zhang, Yan; Wang, Min; Lv, Xiaodan; Wen, Bo; Liu, Hongjun; Luan, Hemi; Zhang, Yonggang; Yang, Shuang; Wang, Xiaodian; Xu, Jiaohui; Li, Xueqin; Li, Shuaicheng; Wang, Junyi; Palloix, Alain; Bosland, Paul W.; Li, Yingrui; Krogh, Anders; Rivera-Bustamante, Rafael F.; Herrera-Estrella, Luis; Yin, Ye; Yu, Jiping; Hu, Kailin; Zhang, Zhiming

    2014-01-01

    As an economic crop, pepper satisfies people’s spicy taste and has medicinal uses worldwide. To gain a better understanding of Capsicum evolution, domestication, and specialization, we present here the genome sequence of the cultivated pepper Zunla-1 (C. annuum L.) and its wild progenitor Chiltepin (C. annuum var. glabriusculum). We estimate that the pepper genome expanded ∼0.3 Mya (with respect to the genome of other Solanaceae) by a rapid amplification of retrotransposons elements, resulting in a genome comprised of ∼81% repetitive sequences. Approximately 79% of 3.48-Gb scaffolds containing 34,476 protein-coding genes were anchored to chromosomes by a high-density genetic map. Comparison of cultivated and wild pepper genomes with 20 resequencing accessions revealed molecular footprints of artificial selection, providing us with a list of candidate domestication genes. We also found that dosage compensation effect of tandem duplication genes probably contributed to the pungent diversification in pepper. The Capsicum reference genome provides crucial information for the study of not only the evolution of the pepper genome but also, the Solanaceae family, and it will facilitate the establishment of more effective pepper breeding programs. PMID:24591624

  7. Ubiquitin Proteasome System in Parkinson Disease: a keeper or a witness?

    Science.gov (United States)

    Martins-Branco, Diogo; Esteves, Ana R.; Santos, Daniel; Arduino, Daniela M.; Swerdlow, Russell H.; Oliveira, Catarina R.; Januario, Cristina; Cardoso, Sandra M.

    2014-01-01

    Objective The aim of this work was to evaluate the role of Ubiquitin-Proteasome System (UPS) on mitochondrial-driven alpha-synuclein (aSN) clearance in in vitro, ex vivo and in vivo Parkinson disease (PD) cellular models. Method We used SH-SY5Y ndufa2 knock-down (KD) cells, PD cybrids and peripheral blood mononuclear cells (PBMC) from patients meeting the diagnostic criteria for PD. We quantified aSN aggregation, proteasome activity and protein ubiquitination levels. In PBMC of PD patients population we evaluated aSN levels in plasma and the influence of several demographic characteristics in the above mentioned determinations. Results We found that ubiquitin-independent proteasome activity was up-regulated in SH-SY5Y ndufa2 KD cells while a down regulation was observed in PD cybrids and PBMC. Moreover, we observed an increase in protein ubiquitination that correlates with a decrease in ubiquitin-dependent proteasome activity. Accordingly, proteasome inhibition prevented ubiquitin-dependent aSN clearance. Ubiquitin-independent proteasome activity was positively correlated with ubiquitination in PBMC. We also report a negative correlation of chymotrypsin-like activity with age in control and late-onset PD groups. Total ubiquitin content is positively correlated with aSN oligomers levels, which leads to an age-dependent increase of aSN ubiquitination in LOPD. Moreover, aSN levels are increased in the plasma of PD patients. Interpretation aSN oligomers are ubiquitinated and we identified an ubiquitin-dependent clearance insufficiency with accumulation of both aSN and ubiquitin. However, SH-SY5Y ndufa2 KD cells showed a significant up-regulation of ubiquitin-independent proteasomal enzymatic activity that could mean a cell rescue attempt. Moreover, we identified that UPS function is age-dependent in PBMC. PMID:22921536

  8. ROS inhibitor N-acetyl-L-cysteine antagonizes the activity of proteasome inhibitors.

    Science.gov (United States)

    Halasi, Marianna; Wang, Ming; Chavan, Tanmay S; Gaponenko, Vadim; Hay, Nissim; Gartel, Andrei L

    2013-09-01

    NAC (N-acetyl-L-cysteine) is commonly used to identify and test ROS (reactive oxygen species) inducers, and to inhibit ROS. In the present study, we identified inhibition of proteasome inhibitors as a novel activity of NAC. Both NAC and catalase, another known scavenger of ROS, similarly inhibited ROS levels and apoptosis associated with H₂O₂. However, only NAC, and not catalase or another ROS scavenger Trolox, was able to prevent effects linked to proteasome inhibition, such as protein stabilization, apoptosis and accumulation of ubiquitin conjugates. These observations suggest that NAC has a dual activity as an inhibitor of ROS and proteasome inhibitors. Recently, NAC was used as a ROS inhibitor to functionally characterize a novel anticancer compound, piperlongumine, leading to its description as a ROS inducer. In contrast, our own experiments showed that this compound depicts features of proteasome inhibitors including suppression of FOXM1 (Forkhead box protein M1), stabilization of cellular proteins, induction of ROS-independent apoptosis and enhanced accumulation of ubiquitin conjugates. In addition, NAC, but not catalase or Trolox, interfered with the activity of piperlongumine, further supporting that piperlongumine is a proteasome inhibitor. Most importantly, we showed that NAC, but not other ROS scavengers, directly binds to proteasome inhibitors. To our knowledge, NAC is the first known compound that directly interacts with and antagonizes the activity of proteasome inhibitors. Taken together, the findings of the present study suggest that, as a result of the dual nature of NAC, data interpretation might not be straightforward when NAC is utilized as an antioxidant to demonstrate ROS involvement in drug-induced apoptosis.

  9. Interaction of the anaphase-promoting complex/cyclosome and proteasome protein complexes with multiubiquitin chain-binding proteins

    DEFF Research Database (Denmark)

    Seeger, Michael; Hartmann-Petersen, Rasmus; Wilkinson, Caroline R M

    2003-01-01

    Fission yeast Rhp23 and Pus1 represent two families of multiubiquitin chain-binding proteins that associate with the proteasome. We show that both proteins bind to different regions of the proteasome subunit Mts4. The binding site for Pus1 was mapped to a cluster of repetitive sequences also found...... in the proteasome subunit SpRpn2 and the anaphase-promoting complex/cyclosome (APC/C) subunit Cut4. The putative role of Pus1 as a factor involved in allocation of ubiquitinylated substrates for the proteasome is discussed....

  10. Dynamic plant uptake model applied for drip irrigation of an insecticide to pepper fruit plants

    DEFF Research Database (Denmark)

    Legind, Charlotte Nielsen; Kennedy, C. M.; Rein, Arno

    2011-01-01

    irrigation, its application for a soil-applied insecticide and a sensitivity analysis of the model parameters. RESULTS: The model predicted the measured increase and decline of residues following two soil applications of an insecticide to peppers, with an absolute error between model and measurement ranging...... from 0.002 to 0.034 mg kg fw—1. Maximum measured concentrations in pepper fruit were approximately 0.22 mg kg fw—1. Temperature was the most sensitive component for predicting the peak and final concentration in pepper fruit, through its influence on soil and plant degradation rates...

  11. The Effect of Red Pepper Waste Added to Compost on Selenium Content of Pleurotus citrinopileatus Singer

    Directory of Open Access Journals (Sweden)

    Fatih Kalyoncu

    2016-03-01

    Full Text Available In this study, selenium levels of P.citrinopileatus added different rates of red pepper waste to compost as a source of selenium were determined. For this purpose, six different compost groups were prepared and inoculated with P.citrinopileatus mycelia. Selenium levels of harvested mushrooms were analysed. In conclusion, 0.338 µg/g selenium were determined in one gram dry mushroom that red pepper waste added to compost at 5%. Mycelia growth and selenium accumulation were negatively affected by higher rates of red pepper waste.

  12. X-band ESR study on evaluation of radicals induced in pasteurized pepper

    International Nuclear Information System (INIS)

    Matsuura, Masaaki; Ogawa, Satoko; Ukai, Mitsuko; Oowada, Shigeru

    2007-01-01

    The radical properties of pasteurized pepper were investigated by means of X-band ESR spectroscopy. Pasteurization process was done by irradiation or steam. There were three radicals in the specimens before and after pasteurization. Upon irradiation a new radical was found. ESR peak intensity of specimen before and after parturition with steam was almost same level. Peak intensity of radiated pepper showed almost 4 times as compare with that of non treated pepper. Radical activity of the specimens after pasteurization showed almost same value. We concluded that radicals were induced by irradiation. But the radical activity was not changed before and after pasteurization. (author)

  13. Proteasome modulator 9 and macrovascular pathology of T2D

    Directory of Open Access Journals (Sweden)

    Gragnoli Claudia

    2011-04-01

    Full Text Available Abstract Aims Coronary artery disease (CAD and stroke share a major linkage at the chromosome 12q24 locus. The same chromosome region entails at least a major risk gene for type 2 diabetes (T2D within NIDDM2, the non-insulin-dependent-diabetes 2 locus. The gene of Proteasome Modulator 9 (PSMD9 lies in the NIDDM2 region and is implicated in diabetes in mice. PSMD9 mutations rarely cause T2D and common variants are linked to both late-onset T2D and maturity-onset-diabetes of the young (MODY3. In this study, we aimed at determining whether PSMD9 is linked to macrovascular pathology of T2D. Methods and Results In our 200 T2D families from Italy, we characterized the clinical phenotype of macrovascular pathology by defining the subjects for presence or absence of CAD, stroke and/or transitory ischemic attacks (TIA, plaques of the large arterial vessels (macro-vasculopathy and arterial angioplasty performance. We then screened 200 T2D siblings/families for PSMD9 +nt460A/G, +nt437C/T and exon E197G A/G single nucleotide polymorphisms (SNPs and performed a non-parametric linkage study to test for linkage for coronary artery disease, stroke/TIA, macro-vasculopathy and macrovascular pathology of T2D. We performed 1,000 replicates to test the power of our significant results. Our results show a consistent significant LOD score in linkage with all the above-mentioned phenotypes. Our 1000 simulation analyses, performed for each single test, confirm that the results are not due to random chance. Conclusions In summary, the PSMD9 IVS3+nt460A/G, +nt437C/T and exon E197G A/G SNPs are linked to CAD, stroke/TIA and macrovascular pathology of T2D in Italians.

  14. The Ubiquitin-Proteasome System and Microvascular Complications of Diabetes

    Directory of Open Access Journals (Sweden)

    Saeed Yadranji Aghdam

    2013-01-01

    Full Text Available The ubiquitin-proteasome system (UPS is the mainstay of protein quality control which regulates cell cycle, differentiation and various signal transduction pathways in eukaryotic cells. The timely and selective degradation of surplus and/or aberrant proteins by the UPS is essential for normal cellular physiology. Any disturbance, delay or exaggeration in the process of selection, sequestration, labeling for degradation and degradation of target proteins by the UPS will compromise cellular and tissue homeostasis. High blood glucose or hyperglycemia caused by diabetes disrupts normal vascular function in several target organs including the retina and kidney resulting in the development of diabetic retinopathy (DR and diabetic nephropathy (DN. We and others have shown that hyperglycemia and oxidative stress modulate UPS activity in the retina and kidney. The majority of studies have focused on the kidney and provided insights into the contribution of dysregulated UPS to microvascular damage in DN. The eye is a unique organ in which a semi-fluid medium, the vitreous humor, separates the neural retina and its anastomosed blood vessels from the semi-solid lens tissue. The complexity of the cellular and molecular components of the eye may require a normal functioning and well tuned UPS for healthy vision. Altered UPS activity may contribute to the development of retinal microvascular complications of diabetes. A better understanding of the molecular nature of the ocular UPS function under normal and diabetic conditions is essential for development of novel strategies targeting its activity. This review will discuss the association of retinal vascular cell UPS activity with microvascular damage in DR with emphasis on alterations of the PA28 subunits of the UPS.

  15. Prospective iterative trial of proteasome inhibitor-based desensitization.

    Science.gov (United States)

    Woodle, E S; Shields, A R; Ejaz, N S; Sadaka, B; Girnita, A; Walsh, R C; Alloway, R R; Brailey, P; Cardi, M A; Abu Jawdeh, B G; Roy-Chaudhury, P; Govil, A; Mogilishetty, G

    2015-01-01

    A prospective iterative trial of proteasome inhibitor (PI)-based therapy for reducing HLA antibody (Ab) levels was conducted in five phases differing in bortezomib dosing density and plasmapheresis timing. Phases included 1 or 2 bortezomib cycles (1.3 mg/m(2) × 6-8 doses), one rituximab dose and plasmapheresis. HLA Abs were measured by solid phase and flow cytometry (FCM) assays. Immunodominant Ab (iAb) was defined as highest HLA Ab level. Forty-four patients received 52 desensitization courses (7 patients enrolled in multiple phases): Phase 1 (n = 20), Phase 2 (n = 12), Phase 3 (n = 10), Phase 4 (n = 5), Phase 5 (n = 5). iAb reductions were observed in 38 of 44 (86%) patients and persisted up to 10 months. In Phase 1, a 51.5% iAb reduction was observed at 28 days with bortezomib alone. iAb reductions increased with higher bortezomib dosing densities and included class I, II, and public antigens (HLA DRβ3, HLA DRβ4 and HLA DRβ5). FCM median channel shifts decreased in 11/11 (100%) patients by a mean of 103 ± 54 mean channel shifts (log scale). Nineteen out of 44 patients (43.2%) were transplanted with low acute rejection rates (18.8%) and de novo DSA formation (12.5%). In conclusion, PI-based desensitization consistently and durably reduces HLA Ab levels providing an alternative to intravenous immune globulin-based desensitization. © Copyright 2014 The American Society of Transplantation and the American Society of Transplant Surgeons.

  16. SOCS-1 localizes to the microtubule organizing complex-associated 20S proteasome.

    Science.gov (United States)

    Vuong, Bao Q; Arenzana, Teresita L; Showalter, Brian M; Losman, Julie; Chen, X Peter; Mostecki, Justin; Banks, Alexander S; Limnander, Andre; Fernandez, Neil; Rothman, Paul B

    2004-10-01

    The regulation of cytokine signaling is critical for controlling cellular proliferation and activation during an immune response. SOCS-1 is a potent inhibitor of Jak kinase activity and of signaling initiated by several cytokines. SOCS-1 protein levels are tightly regulated, and recent data suggest that SOCS-1 may regulate the protein levels of some signaling proteins by the ubiquitin proteasome pathway; however, the cellular mechanism by which SOCS-1 directs proteins for degradation is unknown. In this report, SOCS-1 is found to colocalize and biochemically copurify with the microtubule organizing complex (MTOC) and its associated 20S proteasome. The SOCS-1 SH2 domain is required for the localization of SOCS-1 to the MTOC. Overexpression of SOCS-1 targets Jak1 in an SH2-dependent manner to a perinuclear distribution resembling the MTOC-associated 20S proteasome. Analysis of MTOCs fractionated from SOCS-1-deficient cells demonstrates that SOCS-1 may function redundantly to regulate the localization of Jak1 to the MTOC. Nocodazole inhibits the protein turnover of SOCS-1, demonstrating that the minus-end transport of SOCS-1 to the MTOC-associated 20S proteasome is required to regulate SOCS-1 protein levels. These data link SOCS-1 directly with the proteasome pathway and suggest another function for the SH2 domain of SOCS-1 in the regulation of Jak/STAT signaling.

  17. Proteasome inhibitors block DNA repair and radiosensitize non-small cell lung cancer.

    Directory of Open Access Journals (Sweden)

    Kyle R Cron

    Full Text Available Despite optimal radiation therapy (RT, chemotherapy and/or surgery, a majority of patients with locally advanced non-small cell lung cancer (NSCLC fail treatment. To identify novel gene targets for improved tumor control, we performed whole genome RNAi screens to identify knockdowns that most reproducibly increase NSCLC cytotoxicity. These screens identified several proteasome subunits among top hits, including the topmost hit PSMA1, a component of the core 20 S proteasome. Radiation and proteasome inhibition showed synergistic effects. Proteasome inhibition resulted in an 80-90% decrease in homologous recombination (HR, a 50% decrease in expression of NF-κB-inducible HR genes BRCA1 and FANCD2, and a reduction of BRCA1, FANCD2 and RAD51 ionizing radiation-induced foci. IκBα RNAi knockdown rescued NSCLC radioresistance. Irradiation of mice with NCI-H460 xenografts after inducible PSMA1 shRNA knockdown markedly increased murine survival compared to either treatment alone. Proteasome inhibition is a promising strategy for NSCLC radiosensitization via inhibition of NF-κB-mediated expression of Fanconi Anemia/HR DNA repair genes.

  18. Inhibitors of the proteasome reduce the accelerated proteolysis in atrophying rat skeletal muscles.

    Science.gov (United States)

    Tawa, N E; Odessey, R; Goldberg, A L

    1997-07-01

    Several observations have suggested that the enhanced proteolysis and atrophy of skeletal muscle in various pathological states is due primarily to activation of the ubiquitin-proteasome pathway. To test this idea, we investigated whether peptide aldehyde inhibitors of the proteasome, N-acetyl-leucyl-leucyl-norleucinal (LLN), or the more potent CBZ-leucyl-leucyl-leucinal (MG132) suppressed proteolysis in incubated rat skeletal muscles. These agents (e.g., MG132 at 10 microM) inhibited nonlysosomal protein breakdown by up to 50% (P protein synthesis or amino acid pools, but improved overall protein balance in the muscle. Upon treatment with MG132, ubiquitin-conjugated proteins accumulated in the muscle. The inhibition of muscle proteolysis correlated with efficacy against the proteasome, although these agents could also inhibit calpain-dependent proteolysis induced with Ca2+. These inhibitors had much larger effects on proteolysis in atrophying muscles than in controls. In the denervated soleus undergoing atrophy, the increase in ATP-dependent proteolysis was reduced 70% by MG132 (P muscle proteolysis induced by administering thyroid hormones was reduced 40-70% by the inhibitors. Finally, in rats made septic by cecal puncture, the increase in muscle proteolysis was completely blocked by MG132. Thus, the enhanced proteolysis in many catabolic states (including denervation, hyperthyroidism, and sepsis) is due to a proteasome-dependent pathway, and inhibition of proteasome function may be a useful approach to reduce muscle wasting.

  19. The 20S proteasome splicing activity discovered by SpliceMet.

    Directory of Open Access Journals (Sweden)

    Juliane Liepe

    2010-06-01

    Full Text Available The identification of proteasome-generated spliced peptides (PSP revealed a new unpredicted activity of the major cellular protease. However, so far characterization of PSP was entirely dependent on the availability of patient-derived cytotoxic CD8+ T lymphocytes (CTL thus preventing a systematic investigation of proteasome-catalyzed peptide splicing (PCPS. For an unrestricted PSP identification we here developed SpliceMet, combining the computer-based algorithm ProteaJ with in vitro proteasomal degradation assays and mass spectrometry. By applying SpliceMet for the analysis of proteasomal processing products of four different substrate polypeptides, derived from human tumor as well as viral antigens, we identified fifteen new spliced peptides generated by PCPS either by cis or from two separate substrate molecules, i.e., by trans splicing. Our data suggest that 20S proteasomes represent a molecular machine that, due to its catalytic and structural properties, facilitates the generation of spliced peptides, thereby providing a pool of qualitatively new peptides from which functionally relevant products may be selected.

  20. Mitochondrial Malfunctioning, Proteasome Arrest and Apoptosis in Cancer Cells by Focused Intracellular Generation of Oxygen Radicals

    Directory of Open Access Journals (Sweden)

    Ilaria Postiglione

    2015-08-01

    Full Text Available Photofrin/photodynamic therapy (PDT at sub-lethal doses induced a transient stall in proteasome activity in surviving A549 (p53+/+ and H1299 (p53−/− cells as indicated by the time-dependent decline/recovery of chymotrypsin-like activity. Indeed, within 3 h of incubation, Photofrin invaded the cytoplasm and localized preferentially within the mitochondria. Its light activation determined a decrease in mitochondrial membrane potential and a reversible arrest in proteasomal activity. A similar result is obtained by treating cells with Antimycin and Rotenone, indicating, as a common denominator of this effect, the ATP decrease. Both inhibitors, however, were more toxic to cells as the recovery of proteasomal activity was incomplete. We evaluated whether combining PDT (which is a treatment for killing tumor cells, per se, and inducing proteasome arrest in the surviving ones with Bortezomib doses capable of sustaining the stall would protract the arrest with sufficient time to induce apoptosis in remaining cells. The evaluation of the mitochondrial membrane depolarization, residual proteasome and mitochondrial enzymatic activities, colony-forming capabilities, and changes in protein expression profiles in A549 and H1299 cells under a combined therapeutic regimen gave results consistent with our hypothesis.

  1. Exploiting nature's rich source of proteasome inhibitors as starting points in drug development.

    Science.gov (United States)

    Gräwert, Melissa Ann; Groll, Michael

    2012-02-01

    Cancer is the No. 2 cause of death in the Western world and one of the most expensive diseases to treat. Thus, it is not surprising, that every major pharmaceutical and biotechnology company has a blockbuster oncology product. In 2003, Millennium Pharmaceuticals entered the race with Velcade®, a first-in-class proteasome inhibitor that has been approved by the FDA for treatment of multiple myeloma and its sales have passed the billion dollar mark. Velcade®'s extremely toxic boronic acid pharmacophore, however, contributes to a number of severe side effects. Nevertheless, the launching of this product has validated the proteasome as a target in fighting cancer and further proteasome inhibitors have entered the market as anti-cancer drugs. Additionally, proteasome inhibitors have found application as crop protection agents, anti-parasitics, immunosuppressives, as well as in new therapies for muscular dystrophies and inflammation. Many of these compounds are based on microbial metabolites. In this review, we emphasize the important role of the structural elucidation of the various unique binding mechanisms of these compounds that have been optimized throughout evolution to target the proteasome. Based on this knowledge, medicinal chemists have further optimized these natural products, resulting in potential drugs with reduced off-target activities. This journal is © The Royal Society of Chemistry 2012

  2. Acetylation-Mediated Proteasomal Degradation of Core Histones during DNA Repair and Spermatogenesis

    Science.gov (United States)

    Qian, Min-Xian; Pang, Ye; Liu, Cui Hua; Haratake, Kousuke; Du, Bo-Yu; Ji, Dan-Yang; Wang, Guang-Fei; Zhu, Qian-Qian; Song, Wei; Yu, Yadong; Zhang, Xiao-Xu; Huang, Hai-Tao; Miao, Shiying; Chen, Lian-Bin; Zhang, Zi-Hui; Liang, Ya-Nan; Liu, Shan; Cha, Hwangho; Yang, Dong; Zhai, Yonggong; Komatsu, Takuo; Tsuruta, Fuminori; Li, Haitao; Cao, Cheng; Li, Wei; Li, Guo-Hong; Cheng, Yifan; Chiba, Tomoki; Wang, Linfang; Goldberg, Alfred L.; Shen, Yan; Qiu, Xiao-Bo

    2013-01-01

    SUMMARY Histone acetylation plays critical roles in chromatin remodeling, DNA repair, and epigenetic regulation of gene expression, but the underlying mechanisms are unclear. Proteasomes usually catalyze ATP- and polyubiquitin-dependent proteolysis. Here we show that the proteasomes containing the activator PA200 catalyze the polyubiquitin-independent degradation of histones. Most proteasomes in mammalian testes (“spermatoproteasomes”) contain a spermatid/sperm-specific α-subunit α4s/PSMA8 and/or the catalytic β-subunits of immunoproteasomes in addition to PA200. Deletion of PA200 in mice abolishes acetylation-dependent degradation of somatic core histones during DNA double-strand breaks, and delays core histone disappearance in elongated spermatids. Purified PA200 greatly promotes ATP-independent proteasomal degradation of the acetylated core histones, but not polyubiquitinated proteins. Furthermore, acetylation on histones is required for their binding to the bromodomain-like regions in PA200 and its yeast ortholog, Blm10. Thus, PA200/Blm10 specifically targets the core histones for acetylation-mediated degradation by proteasomes, providing mechanisms by which acetylation regulates histone degradation, DNA repair, and spermatogenesis. PMID:23706739

  3. Repression of protein translation and mTOR signaling by proteasome inhibitor in colon cancer cells

    International Nuclear Information System (INIS)

    Wu, William Ka Kei; Volta, Viviana; Cho, Chi Hin; Wu, Ya Chun; Li, Hai Tao; Yu, Le; Li, Zhi Jie; Sung, Joseph Jao Yiu

    2009-01-01

    Protein homeostasis relies on a balance between protein synthesis and protein degradation. The ubiquitin-proteasome system is a major catabolic pathway for protein degradation. In this respect, proteasome inhibition has been used therapeutically for the treatment of cancer. Whether inhibition of protein degradation by proteasome inhibitor can repress protein translation via a negative feedback mechanism, however, is unknown. In this study, proteasome inhibitor MG-132 lowered the proliferation of colon cancer cells HT-29 and SW1116. In this connection, MG-132 reduced the phosphorylation of mammalian target of rapamycin (mTOR) at Ser2448 and Ser2481 and the phosphorylation of its downstream targets 4E-BP1 and p70/p85 S6 kinases. Further analysis revealed that MG-132 inhibited protein translation as evidenced by the reductions of 35 S-methionine incorporation and polysomes/80S ratio. Knockdown of raptor, a structural component of mTOR complex 1, mimicked the anti-proliferative effect of MG-132. To conclude, we demonstrate that the inhibition of protein degradation by proteasome inhibitor represses mTOR signaling and protein translation in colon cancer cells.

  4. Comparative genomics reveals diversity among xanthomonads infecting tomato and pepper

    LENUS (Irish Health Repository)

    Potnis, Neha

    2011-03-11

    Abstract Background Bacterial spot of tomato and pepper is caused by four Xanthomonas species and is a major plant disease in warm humid climates. The four species are distinct from each other based on physiological and molecular characteristics. The genome sequence of strain 85-10, a member of one of the species, Xanthomonas euvesicatoria (Xcv) has been previously reported. To determine the relationship of the four species at the genome level and to investigate the molecular basis of their virulence and differing host ranges, draft genomic sequences of members of the other three species were determined and compared to strain 85-10. Results We sequenced the genomes of X. vesicatoria (Xv) strain 1111 (ATCC 35937), X. perforans (Xp) strain 91-118 and X. gardneri (Xg) strain 101 (ATCC 19865). The genomes were compared with each other and with the previously sequenced Xcv strain 85-10. In addition, the molecular features were predicted that may be required for pathogenicity including the type III secretion apparatus, type III effectors, other secretion systems, quorum sensing systems, adhesins, extracellular polysaccharide, and lipopolysaccharide determinants. Several novel type III effectors from Xg strain 101 and Xv strain 1111 genomes were computationally identified and their translocation was validated using a reporter gene assay. A homolog to Ax21, the elicitor of XA21-mediated resistance in rice, and a functional Ax21 sulfation system were identified in Xcv. Genes encoding proteins with functions mediated by type II and type IV secretion systems have also been compared, including enzymes involved in cell wall deconstruction, as contributors to pathogenicity. Conclusions Comparative genomic analyses revealed considerable diversity among bacterial spot pathogens, providing new insights into differences and similarities that may explain the diverse nature of these strains. Genes specific to pepper pathogens, such as the O-antigen of the lipopolysaccharide cluster

  5. Effect of the smoking process and firewood type in the phytochemical content and antioxidant capacity of red Jalapeño pepper during its transformation to chipotle pepper.

    Science.gov (United States)

    Moreno-Escamilla, Jesús Omar; de la Rosa, Laura A; López-Díaz, José Alberto; Rodrigo-García, Joaquín; Núñez-Gastélum, José Alberto; Alvarez-Parrilla, Emilio

    2015-10-01

    Chipotle pepper is a dry smoked red Jalapeño pepper well appreciated in the Mexican and American cuisines. Phytochemical content and antioxidant activity of red Jalapeño pepper has been previously reported, however, the effect of the smoking process in the content of phytochemicals, antioxidant capacity and capsaicinoids has not been previously reported. In the present study we found that the smoking process had a significant effect on the content of polyphenolic compounds and antioxidant capacity of red Jalapeño pepper. It induced a nearly 50% increase in total phenols and 15% increase in antioxidant capacity in chipotle pepper compared to fresh samples (in dry weight basis). Capsaicinoids and ascorbic acid content decreased in smoked samples, while carotenoids remained practically unchanged. The strongest effect of smoking was observed in the content of total flavonoids and catechin determined by HPLC. Therefore we can conclude that smoking process has a positive effect in the fruit because it raises its phytochemical properties. The type of wood used in the smoking process also had an effect on the phytochemical content and antioxidant capacity with traditional pecan wood being the best. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Detection of organic free radicals in irradiated pepper by electron spin resonance

    International Nuclear Information System (INIS)

    Ukai, Mitsuko; Shimoyama, Yuhei

    2002-01-01

    Using electron spin resonance (ESR) spectroscopy, we revealed various free radicals in a Japanese commercially available black pepper before and after γ-irradiation. The representative ESR spectrum of the pepper is composed of a sextet centered at g=2.0, a singlet at the same g-value and a singlet at g=4.0. The first one is attributable to a signal with hyperfine interactions of Mn 2+ ion (7.4 mT). The second one is due to an organic free radical. The third one may be originated from Fe 3+ ion of the non-hem Fe in proteins. A pair of signals appeared in the black pepper after γ-irradiation. The progressive saturation behavior reconfirmed the signal identification for the radicals in the black pepper. (author)

  7. Effect of sweet pepper cultivation on the content of phytotoxic phenolic compounds in substrates

    Directory of Open Access Journals (Sweden)

    Barbara Politycka

    2013-12-01

    Full Text Available The aim of the conducted study was to determine to what extent the cultivation of sweet pepper lowers the phytotoxicity of a substrate. The examined material was a highly phytotoxic substrate due to repeated cucumber growing on it. This substrate was a mixture of pine and beech bark, low peat and sawdust. Five sweet pepper cultivars: Amador, Bell Boy, Culinar, Poznańska Słodka and WSE 2/82 were planted. During vegetation of the sweet pepper phytotoxicity and phenolics levels were determined in the substrate. It was found that cultivation of sweet pepper had a significant effect on lowering phytotoxicity and phenolics content in the substrate. Among the five tested cultivars, the highest detoxicating ability was exhibited by Amador, Culinar and WSE 2/82 while the Poznańska Słodka did not show such abilities.

  8. Pepper-pot diagnostic method to define emittance and Twiss parameters on low energies accelerators

    CERN Document Server

    Dolinska, M E

    2002-01-01

    The new complex mathematical algorithm to determine beam transverse emittance data and the Twiss parameters from intensity measured with pepper-por diagnostic device on rf low energies accelerators is described.

  9. A new environment-friendly hot pepper variety "Shiyan No. 1"

    Science.gov (United States)

    Han, Jianming; Xu, Shuzhen; Wang, Ruiling; Zhang, Yanzhao; Yun, Chao

    2018-04-01

    Hot pepper has rich genetic diversity which is the important base of breeding of new variety, and it is also one of the important vegetable in the word. In this study, we bred the "Shiyan No. 1" environment-friendly hot pepper variety using hybrid method on the basis of hot pepper genetic diversity. "Shiyan No. 1" is a new F1 hybrid of hot pepper variety with mid-early maturity. The new variety has a high productivity of 3000-5000kg(667m2)-1 with thick oxhorn shape fruits, green skin, thick flesh, mild-hot taste, good quality and marketable characters. It can reduce chemical pesticides usage and thereby protect environment because it is resistant to virus disease, highly resistant to phytophthora blight, anthracnose and bacterial wilt. In conclusion, the new bred "Shiyan No.1" is suitable for protected cultivation and open field cultivation in China.

  10. Physiological, Biochemical and Genetic Characteristics of Ralstonia solanacearum Strains Isolated from Pepper Plants in Korea

    Directory of Open Access Journals (Sweden)

    Young Kee Lee

    2013-12-01

    Full Text Available Totally sixty three bacteria were isolated from lower stems showing symptoms of bacterial wilt on pepper plants in 14 counties of 7 provinces, Korea. The isolates showed strong pathogenicity on red pepper (cv. Daewang and tomato (cv. Seogwang seedlings. All virulent bacteria were identified as Ralstonia solanacearum based on colony types, physiological and biochemical tests and polymerase chain reaction (PCR. All R. solanacearum isolates from peppers were race 1. The bacterial isolates consisted of biovar 3 (27% and biovar 4 (73%. Based on polymorphic PCR bands generated by repetitive sequence (rep-PCR, the 63 R. solanacearum isolates were divided into 12 groups at 70% similarity level. These results will be used as basic materials for resistant breeding program and efficient control against bacterial wilt disease of pepper.

  11. Deficiency symptoms of nitrogen, phosphorus, calcium and sulphur in pepper (Capsicum spp) cv malagueta

    International Nuclear Information System (INIS)

    Maruca, F.A.; Gouveia, P.R.; Teixeira, N.T.

    1989-01-01

    The pepper (Capsium spp) was cultivated in sand and nutritive solution to describe the deficiency symptoms of N, P, Ca and S. The deficiency symptoms and the first lack signals were clear to P and N nutrient. (author) [pt

  12. Sensory properties of chile pepper heat - and its importance to food quality and cultural preference.

    Science.gov (United States)

    Guzmán, Ivette; Bosland, Paul W

    2017-10-01

    Chile peppers are one of the most important vegetable and spice crops in the world. They contain capsaicinoids that are responsible for the characteristic burning (pungency) sensation. Currently, there are 22 known naturally occurring capsaicinoids that can cause the heat sensation when consumed. Each produces a different heat sensation effect in the mouth. A need exists for a standard and new terminology to describe the complex heat sensation one feels when eating a chile pepper. A comprehensive set of descriptors to describe the sensory characteristics of chile pepper heat was developed. It was validated with trained panelists tasting samples representing the five domesticated species and 14 pod-types within these species. Five key attributes that define and reference a lexicon for describing the heat sensation of chile peppers were determined to describe the heat sensation in any product, and importantly, can be used in the food industry worldwide. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  13. Identification of Optimal Reference Genes for Normalization of qPCR Analysis during Pepper Fruit Development

    Directory of Open Access Journals (Sweden)

    Yuan Cheng

    2017-06-01

    Full Text Available Due to its high sensitivity and reproducibility, quantitative real-time PCR (qPCR is practiced as a useful research tool for targeted gene expression analysis. For qPCR operations, the normalization with suitable reference genes (RGs is a crucial step that eventually determines the reliability of the obtained results. Although pepper is considered an ideal model plant for the study of non-climacteric fruit development, at present no specific RG have been developed or validated for the qPCR analyses of pepper fruit. Therefore, this study aimed to identify stably expressed genes for their potential use as RGs in pepper fruit studies. Initially, a total of 35 putative RGs were selected by mining the pepper transcriptome data sets derived from the PGP (Pepper Genome Platform and PGD (Pepper Genome Database. Their expression stabilities were further measured in a set of pepper (Capsicum annuum L. var. 007e fruit samples, which represented four different fruit developmental stages (IM: Immature; MG: Mature green; B: Break; MR: Mature red using the qPCR analysis. Then, based on the qPCR results, three different statistical algorithms, namely geNorm, Normfinder, and boxplot, were chosen to evaluate the expression stabilities of these putative RGs. It should be noted that nine genes were proven to be qualified as RGs during pepper fruit development, namely CaREV05 (CA00g79660; CaREV08 (CA06g02180; CaREV09 (CA06g05650; CaREV16 (Capana12g002666; CaREV21 (Capana10g001439; CaREV23 (Capana05g000680; CaREV26 (Capana01g002973; CaREV27 (Capana11g000123; CaREV31 (Capana04g002411; and CaREV33 (Capana08g001826. Further analysis based on geNorm suggested that the application of the two most stably expressed genes (CaREV05 and CaREV08 would provide optimal transcript normalization in the qPCR experiments. Therefore, a new and comprehensive strategy for the identification of optimal RGs was developed. This strategy allowed for the effective normalization of the q

  14. Regulation of SUMO2 Target Proteins by the Proteasome in Human Cells Exposed to Replication Stress

    DEFF Research Database (Denmark)

    Bursomanno, Sara; McGouran, Joanna F; Kessler, Benedikt M

    2015-01-01

    In human cells, SUMO2 is predominantly conjugated to target proteins in response to cellular stress. Previous studies suggested that proteins conjugated to SUMO2, but not to SUMO1, could be regulated by the ubiquitin-mediated proteasome system. Hence, we set out to understand the role...... of the proteasome in determining the fate of proteins conjugated to SUMO2 when cells are treated with DNA replication stress conditions. We conducted a quantitative proteomic analysis in a U2OS cell line stably expressing SUMO2(Q87R) tagged with StrepHA in the presence or absence of epoxomicin (EPOX), a proteasome...... inhibitor. We identified subgroups of putative SUMO2 targets that were either degraded or stabilized by EPOX upon SUMO2 conjugation in response to replication stress. Interestingly, the subgroup of proteins degraded upon SUMO2 conjugation was enriched in proteins playing roles in DNA damage repair...

  15. Therapeutic Strategies against Epstein-Barr Virus-Associated Cancers Using Proteasome Inhibitors.

    Science.gov (United States)

    Hui, Kwai Fung; Tam, Kam Pui; Chiang, Alan Kwok Shing

    2017-11-21

    Epstein-Barr virus (EBV) is closely associated with several lymphomas (endemic Burkitt lymphoma, Hodgkin lymphoma and nasal NK/T-cell lymphoma) and epithelial cancers (nasopharyngeal carcinoma and gastric carcinoma). To maintain its persistence in the host cells, the virus manipulates the ubiquitin-proteasome system to regulate viral lytic reactivation, modify cell cycle checkpoints, prevent apoptosis and evade immune surveillance. In this review, we aim to provide an overview of the mechanisms by which the virus manipulates the ubiquitin-proteasome system in EBV-associated lymphoid and epithelial malignancies, to evaluate the efficacy of proteasome inhibitors on the treatment of these cancers and discuss potential novel viral-targeted treatment strategies against the EBV-associated cancers.

  16. The putative roles of the ubiquitin/proteasome pathway in resistance to anticancer therapy.

    Science.gov (United States)

    Smith, Laura; Lind, Michael J; Drew, Philip J; Cawkwell, Lynn

    2007-11-01

    The ubiquitin/proteasome (UP) pathway plays a significant role in many important biological functions and alterations in this pathway have been shown to contribute to the pathology of many human diseases, including cancer. Proteasome inhibition has been well established as a rational strategy for the treatment of multiple myeloma and is currently under investigation for the treatment of other haematological malignancies and solid tumours. Recent evidence suggests that proteasome inhibition may also sensitise tumour cells to the actions of both conventional chemotherapy and radiotherapy, suggesting that this pathway may modify clinical response to anticancer therapy. However, conflicting evidence exists as to the roles of the UP pathway in resistance to treatment. This review endeavours to discuss such roles.

  17. Proteomic analysis of MG132-treated germinating pollen reveals expression signatures associated with proteasome inhibition.

    Directory of Open Access Journals (Sweden)

    Candida Vannini

    Full Text Available Chemical inhibition of the proteasome has been previously found to effectively impair pollen germination and tube growth in vitro. However, the mediators of these effects at the molecular level are unknown. By performing 2DE proteomic analysis, 24 differentially expressed protein spots, representing 14 unique candidate proteins, were identified in the pollen of kiwifruit (Actinidia deliciosa germinated in the presence of the MG132 proteasome inhibitor. qPCR analysis revealed that 11 of these proteins are not up-regulated at the mRNA level, but are most likely stabilized by proteasome inhibition. These differentially expressed proteins are predicted to function in various pathways including energy and lipid metabolism, cell wall synthesis, protein synthesis/degradation and stress responses. In line with this evidence, the MG132-induced changes in the proteome were accompanied by an increase in ATP and ROS content and by an alteration in fatty acid composition.

  18. Sensitive detection of proteasomal activation using the Deg-On mammalian synthetic gene circuit.

    Science.gov (United States)

    Zhao, Wenting; Bonem, Matthew; McWhite, Claire; Silberg, Jonathan J; Segatori, Laura

    2014-04-08

    The ubiquitin proteasome system (UPS) has emerged as a drug target for diverse diseases characterized by altered proteostasis, but pharmacological agents that enhance UPS activity have been challenging to establish. Here we report the Deg-On system, a genetic inverter that translates proteasomal degradation of the transcriptional regulator TetR into a fluorescent signal, thereby linking UPS activity to an easily detectable output, which can be tuned using tetracycline. We demonstrate that this circuit responds to modulation of UPS activity in cell culture arising from the inhibitor MG-132 and activator PA28γ. Guided by predictive modelling, we enhanced the circuit's signal sensitivity and dynamic range by introducing a feedback loop that enables self-amplification of TetR. By linking UPS activity to a simple and tunable fluorescence output, these genetic inverters will enable a variety of applications, including screening for UPS activating molecules and selecting for mammalian cells with different levels of proteasome activity.

  19. Tocopherol synthesis from homogentisate in Capsicum anuum L. (yellow pepper) chromoplast membranes: evidence for tocopherol cyclase.

    OpenAIRE

    Arango, Y; Heise, K P

    1998-01-01

    The present study shows for the first time appreciable tocopherol cyclase activities in plastidial membrane preparations of Capsicum annuum L. (yellow pepper) fruits. When chromoplast membranes from yellow peppers were incubated with [3H]homogentisate and phytyl pyrophosphate under strictly reducing conditions, all biosynthesis precursors were labelled. The main labelling was found in gamma-tocopherol. These observations contradict the hypothesis that assigns a rate-limiting function to tocop...

  20. An l1-TV Algorithm for Deconvolution with Salt and Pepper Noise

    Science.gov (United States)

    2009-04-01

    deblurring in the presence of impulsive noise ,” Int. J. Comput. Vision, vol. 70, no. 3, pp. 279–298, Dec. 2006. [13] A. E. Beaton and J. W. Tukey, “The...AN 1-TV ALGORITHM FOR DECONVOLUTIONWITH SALT AND PEPPER NOISE Brendt Wohlberg∗ T-7 Mathematical Modeling and Analysis Los Alamos National Laboratory...and pepper noise , but the extension of this formulation to more general prob- lems, such as deconvolution, has received little attention. We consider

  1. Metabolite Profiling of Peppers of Various Colors Reveals Relationships Between Tocopherol, Carotenoid, and Phytosterol Content.

    Science.gov (United States)

    Kim, Tae Jin; Choi, Jaehyuk; Kim, Kil Won; Ahn, Soon Kil; Ha, Sun-Hwa; Choi, Yongsoo; Park, Nam Il; Kim, Jae Kwang

    2017-12-01

    Peppers are widely consumed in Korea; the varietal development of peppers with increased content of beneficial plant metabolites is, therefore, of considerable interest. This requires a comprehensive understanding of the metabolic profile of pepper plants and the factors affecting this profile. To this end, we determined the content of various metabolites, such as hydrophilic and lipophilic compounds, phenolic acids, carotenoids, and capsaicinoids in peppers of various colors (green, red, pale green, and violet peppers) and in a high-pungency (green) pepper. We also performed principal component analysis (PCA), Pearson's correlation analysis, and hierarchical clustering analysis (HCA) to determine the relationships among these metabolites in peppers. PCA results indicated no significant variances among the 3 sample replicates. The HCA showed correlations between the metabolites resulting from common or closely linked biosynthesis pathways. Our results showed that carotenoids correlated positively with tocopherols and negatively with phytosterols; our findings also indicated a close relationship between the methylerythritol 4-phosphate and mevalonic acid biosynthesis pathways, providing evidence in favor of an earlier hypothesis regarding crosstalk across the chloroplast membrane. We, thus, demonstrate that metabolic profiling combined with multivariate analysis is a useful tool for analyzing metabolic networks. A total of 71 metabolites were measured in 5 peppers of different colors. The metabolic profiling with multivariate analysis revealed that tocopherol content had a positive correlation with the carotenoid content and a negative correlation with the phytosterol content. The results of this study may help in breeding programs to produce new germplasm with enhanced nutritional quality. © 2017 Institute of Food Technologists®.

  2. Cause of the Scion Death in Green Pepper Grafting System by a Tobamovirus

    Directory of Open Access Journals (Sweden)

    Gug-Seoun Choi

    2011-08-01

    Full Text Available This experiment was attempted to investigate a cause of the scion death in green pepper grafting system. A tobamovirus particle examined in the rootstock of the sample but not in the scion showing necrosis. The virus isolated from the rootstock was identified as Pepper mild mottle virus (PMMoV, pepper tobamovirus pathotype P1.2. (PMMoV-2, by nucleotide sequence analysis and host plant reaction. The virus isolate infected systematically in 6 commercial rootstock varieties using for green pepper grafting seedling production. Green pepper varieties ``Long green mart`` and ``Daechan`` represented resistance to the virus showing local lesions only on the inoculated leaves and ``Manitda`` was systematically infected. In the experiment with grafting ``Long green mart`` or ``Daechan`` onto the those rootstocks, the upper leaves of the scions first showed vein necrosis and wilt symptoms 7 days after inoculation with PMMoV-2 on the cotyledon of the rootstock, following to the scion stem necrosis and then only the scion death. The virus was detected in the rootstock but not in the scion. However, ``Manitda`` of susceptible variety in the grafting system showed mottle symptom on the leaves of the scion but not necrosis on the plant. PMMoV-3 isolate, pepper tobamovirus pathotype P1.2.3, did not cause the scion death in the grafting system. All of the varieties were susceptible to PMMoV-3. These results suggest that the scion death is caused by infecting with pepper tobamovirus pathotype P1.2. in the green pepper grafting system combined with the susceptible rootstock and the resistance scion to the virus pathotype.

  3. Xanthomonas euvesicatoria Causes Bacterial Spot Disease on Pepper Plant in Korea

    Directory of Open Access Journals (Sweden)

    Min-Seong Kyeon

    2016-10-01

    Full Text Available In 2004, bacterial spot-causing xanthomonads (BSX were reclassified into 4 species—Xanthomonas euvesicatoria, X. vesicatoria, X. perforans, and X. gardneri. Bacterial spot disease on pepper plant in Korea is known to be caused by both X. axonopodis pv. vesicatoria and X. vesicatoria. Here, we reidentified the pathogen causing bacterial spots on pepper plant based on the new classification. Accordingly, 72 pathogenic isolates were obtained from the lesions on pepper plants at 42 different locations. All isolates were negative for pectolytic activity. Five isolates were positive for amylolytic activity. All of the Korean pepper isolates had a 32 kDa-protein unique to X. euvesicatoria and had the same band pattern of the rpoB gene as that of X. euvesicatoria and X. perforans as indicated by PCR-restriction fragment length polymorphism analysis. A phylogenetic tree of 16S rDNA sequences showed that all of the Korean pepper plant isolates fit into the same group as did all the reference strains of X. euvesicatoria and X. perforans. A phylogenetic tree of the nucleotide sequences of 3 housekeeping genes—gapA, gyrB, and lepA showed that all of the Korean pepper plant isolates fit into the same group as did all of the references strains of X. euvesicatoria. Based on the phenotypic and genotypic characteristics, we identified the pathogen as X. euvesicatoria. Neither X. vesicatoria, the known pathogen of pepper bacterial spot, nor X. perforans, the known pathogen of tomato plant, was isolated. Thus, we suggest that the pathogen causing bacterial spot disease of pepper plants in Korea is X. euvesicatoria.

  4. Foliar Application of the Fungicide Pyraclostrobin Reduced Bacterial Spot Disease of Pepper

    OpenAIRE

    Beom Ryong Kang; Jang Hoon Lee; Young Cheol Kim

    2018-01-01

    Pyraclostrobin is a broad-spectrum fungicide that inhibits mitochondrial respiration. However, it may also induce systemic resistance effective against bacterial and viral diseases. In this study, we evaluated whether pyraclostrobin enhanced resistance against the bacterial spot pathogen, Xanthomonas euvesicatora on pepper (Capsicum annuum). Although pyraclostrobin alone did not suppressed the in vitro growth of X. euvesicatoria, disease severity in pepper was significantly lower by 69% after...

  5. Denervation-Induced Activation of the Ubiquitin-Proteasome System Reduces Skeletal Muscle Quantity Not Quality.

    Science.gov (United States)

    Baumann, Cory W; Liu, Haiming M; Thompson, LaDora V

    2016-01-01

    It is well known that the ubiquitin-proteasome system is activated in response to skeletal muscle wasting and functions to degrade contractile proteins. The loss of these proteins inevitably reduces skeletal muscle size (i.e., quantity). However, it is currently unknown whether activation of this pathway also affects function by impairing the muscle's intrinsic ability to produce force (i.e., quality). Therefore, the purpose of this study was twofold, (1) document how the ubiquitin-proteasome system responds to denervation and (2) identify the physiological consequences of these changes. To induce soleus muscle atrophy, C57BL6 mice underwent tibial nerve transection of the left hindlimb for 7 or 14 days (n = 6-8 per group). At these time points, content of several proteins within the ubiquitin-proteasome system were determined via Western blot, while ex vivo whole muscle contractility was specifically analyzed at day 14. Denervation temporarily increased several key proteins within the ubiquitin-proteasome system, including the E3 ligase MuRF1 and the proteasome subunits 19S, α7 and β5. These changes were accompanied by reductions in absolute peak force and power, which were offset when expressed relative to physiological cross-sectional area. Contrary to peak force, absolute and relative forces at submaximal stimulation frequencies were significantly greater following 14 days of denervation. Taken together, these data represent two keys findings. First, activation of the ubiquitin-proteasome system is associated with reductions in skeletal muscle quantity rather than quality. Second, shortly after denervation, it appears the muscle remodels to compensate for the loss of neural activity via changes in Ca2+ handling.

  6. Diclofenac induces proteasome and mitochondrial dysfunction in murine cardiomyocytes and hearts.

    Science.gov (United States)

    Ghosh, Rajeshwary; Goswami, Sumanta K; Feitoza, Luis Felipe B B; Hammock, Bruce; Gomes, Aldrin V

    2016-11-15

    One of the most common nonsteroidal anti-inflammatory drugs (NSAIDs) used worldwide, diclofenac (DIC), has been linked to increased risk of cardiovascular disease (CVD). The molecular mechanism(s) by which DIC causes CVD is unknown. Proteasome activities were studied in hearts, livers, and kidneys from male Swiss Webster mice treated with either 100mg/kg DIC for 18h (acute treatment) or 10mg/kg DIC for 28days (chronic treatment). Cultured H9c2 cells and neonatal cardiomyocytes were also treated with different concentrations of DIC and proteasome function, cell death and ROS generation studied. Isolated mouse heart mitochondria were utilized to determine the effect of DIC on various electron transport chain complex activities. DIC significantly inhibited the chymotrypsin-like proteasome activity in rat cardiac H9c2 cells, murine neonatal cardiomyocytes, and mouse hearts, but did not affect proteasome subunit expression levels. Proteasome activity was also affected in liver and kidney tissues from DIC treated animals. The levels of polyubiquitinated proteins increased in hearts from DIC treated mice. Importantly, the levels of oxidized proteins increased while the β5i immunoproteasome activity decreased in hearts from DIC treated mice. DIC increased ROS production and cell death in H9c2 cells and neonatal cardiomyocytes while the cardioprotective NSAID, aspirin, had no effect on ROS levels or cell viability. DIC inhibited mitochondrial Complex III, a major source of ROS, and impaired mitochondrial membrane potential suggesting that mitochondria are the major sites of ROS generation. These results suggest that DIC induces cardiotoxicity by a ROS dependent mechanism involving mitochondrial and proteasome dysfunction. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  7. Comparison between apparent viscosity related to irradiation dose for corn starch and black pepper

    International Nuclear Information System (INIS)

    Casandroiu, T.; Oprita, N.; Ferdes, O.S.

    1999-01-01

    Dose-effect relationship was studied in the rheoviscometric behaviour of geliffied suspensions of irradiated corn starch and black pepper, as the variation of the apparent viscosity and the shear stress related to the dose. Irradiation has been performed up to 16 kGy. Black pepper was ground and sieved to three particle sizes to analyse also the influence of particle size on the apparent viscosity variation by dose. The rheoviscometric measurements have been carried out by a rotationary viscometer on geliffied suspensions of starch and black pepper, into equivalent starch concentration and alkalinised suspensions for pepper. For starch, shear stress variation by dose is exponential, where the coefficients depend on the shear rate. For black pepper, the curves of apparent viscosity relation to dose also fit an exponential equation and the influence of particle size is discussed, too. Viscometric behaviour similar to irradiation of both corn starch and black pepper could be attributed to starch degradation at relatively high doses and should be used to develop an identification and control method for the ionizing treatment of starch-based food materials. (author)

  8. Influence of combinations of fenugreek, garlic, and black pepper powder on production traits of the broilers

    Science.gov (United States)

    Kirubakaran, A.; Moorthy, M.; Chitra, R.; Prabakar, G.

    2016-01-01

    Aim: To study the effects of combinations of fenugreek (Trigonella foenum-graecum L.), garlic (Allium sativum), and black pepper (Piper nigrum) powder supplementation on production traits of broiler chickens. Materials and Methods: A total of 288 commercial broiler chicks were randomly assigned to 1-9 groups with 4 replicates each. An experiment was conducted in broilers with different feed formulations; control feed, with no added fenugreek, garlic, and black pepper powder; and 8 treatment groups receiving feed supplemented with different combinations of fenugreek, garlic, and black pepper powder. The individual broilers’ body weight and feed consumption were recorded and calculate the body weight gain and feed conversion ratio (FCR). Results: Broiler’s weight gain and FCR were significantly higher in groups receiving feed supplemented with garlic and black pepper powder combinations (ppepper powder combinations (ppepper powder supplemented broiler feed fed groups showed higher production performance. The 5 g/kg garlic powder+1 g/kg black pepper powder and 10 g/kg garlic powder+2 g/kg black pepper powder significantly improved the weight gain and FCR. PMID:27284222

  9. Hydrodynamics study on drying of pepper in swirling fluidized bed dryer (SFBD)

    Science.gov (United States)

    Syaif Haron, Nazrul; Hazri Zakaria, Jamal; Faizal Mohideen Batcha, Mohd

    2017-08-01

    Malaysia is one of the pepper producer with exports quantity reaching more than 90000 tonnes between 2010 until 2016. Drying of pepper is mandatory before their export and at present, pepper was dried by sun drying to reduce cost. This conventional drying method was time consuming and may take four days during rainy season, which retards the production of pepper. This paper proposes the swirling fluidized bed drying (SFBD) method, which was known to have high mixing ability and improved solid-gas contact to shorten the drying time of products. A lab scale SFBD system was constructed to carry out this study. Hydrodynamic study was conducted for three beds loadings of 1.0 kg, 1.4 kg at a drying temperature of 90°C. The SFBD has shown excellent potential to dry the pepper with a relatively short drying time compared to the conventional method. Batch drying for the bed loads studied only took 3 hours of drying time only. It was found that bed higher bed loading of wet pepper requires longer drying time due to higher amount of moisture content in the bed. Four distinct regimes of operation were found during drying in the SFBD and these regimes offer flexibility of operation. The total bed pressure drop was relatively low during drying.

  10. Color and illuminance level of lighting can modulate willingness to eat bell peppers.

    Science.gov (United States)

    Hasenbeck, Aimee; Cho, Sungeun; Meullenet, Jean-François; Tokar, Tonya; Yang, Famous; Huddleston, Elizabeth A; Seo, Han-Seok

    2014-08-01

    Food products are often encountered under colored lighting, particularly in restaurants and retail stores. However, relatively little attention has been paid to whether the color of ambient lighting can affect consumers' motivation for consumption. This study aimed to determine whether color (Experiment 1) and illuminance level (Experiment 2) of lighting can influence consumers' liking of appearance and their willingness to eat bell peppers. For red, green, and yellow bell peppers, yellow and blue lighting conditions consistently increased participants' liking of appearance the most and the least, respectively. Participants' willingness to consume bell peppers increased the most under yellow lighting and the least under blue lighting. In addition, a dark condition (i.e. low level of lighting illuminance) decreased liking of appearance and willingness to eat the bell peppers compared to a bright condition (i.e. high level of lighting illuminance). Our findings demonstrate that lighting color and illuminance level can influence consumers' hedonic impression and likelihood to consume bell peppers. Furthermore, the influences of color and illuminance level of lighting appear to be dependent on the surface color of bell peppers. © 2013 Society of Chemical Industry.

  11. Biochemistry and Molecular Biology of Carotenoid Biosynthesis in Chili Peppers (Capsicum spp.

    Directory of Open Access Journals (Sweden)

    María del Rocío Gómez-García

    2013-09-01

    Full Text Available Capsicum species produce fruits that synthesize and accumulate carotenoid pigments, which are responsible for the fruits’ yellow, orange and red colors. Chili peppers have been used as an experimental model for studying the biochemical and molecular aspects of carotenoid biosynthesis. Most reports refer to the characterization of carotenoids and content determination in chili pepper fruits from different species, cultivars, varieties or genotypes. The types and levels of carotenoids differ between different chili pepper fruits, and they are also influenced by environmental conditions. Yellow-orange colors of chili pepper fruits are mainly due to the accumulation of α- and β-carotene, zeaxanthin, lutein and β-cryptoxanthin. Carotenoids such as capsanthin, capsorubin and capsanthin-5,6-epoxide confer the red colors. Chromoplasts are the sites of carotenoid pigment synthesis and storage. According to the most accepted theory, the synthesis of carotenoids in chili peppers is controlled by three loci: c1, c2 and y. Several enzymes participating in carotenoid biosynthesis in chili pepper fruits have been isolated and characterized, and the corresponding gene sequences have been reported. However, there is currently limited information on the molecular mechanisms that regulate this biosynthetic pathway. Approaches to gain more knowledge of the regulation of carotenoid biosynthesis are discussed.

  12. Biochemistry and Molecular Biology of Carotenoid Biosynthesis in Chili Peppers (Capsicum spp.)

    Science.gov (United States)

    del Rocío Gómez-García, María; Ochoa-Alejo, Neftalí

    2013-01-01

    Capsicum species produce fruits that synthesize and accumulate carotenoid pigments, which are responsible for the fruits’ yellow, orange and red colors. Chili peppers have been used as an experimental model for studying the biochemical and molecular aspects of carotenoid biosynthesis. Most reports refer to the characterization of carotenoids and content determination in chili pepper fruits from different species, cultivars, varieties or genotypes. The types and levels of carotenoids differ between different chili pepper fruits, and they are also influenced by environmental conditions. Yellow-orange colors of chili pepper fruits are mainly due to the accumulation of α- and β-carotene, zeaxanthin, lutein and β-cryptoxanthin. Carotenoids such as capsanthin, capsorubin and capsanthin-5,6-epoxide confer the red colors. Chromoplasts are the sites of carotenoid pigment synthesis and storage. According to the most accepted theory, the synthesis of carotenoids in chili peppers is controlled by three loci: c1, c2 and y. Several enzymes participating in carotenoid biosynthesis in chili pepper fruits have been isolated and characterized, and the corresponding gene sequences have been reported. However, there is currently limited information on the molecular mechanisms that regulate this biosynthetic pathway. Approaches to gain more knowledge of the regulation of carotenoid biosynthesis are discussed. PMID:24065101

  13. Real-Time Polymerase Chain Reaction Quantification of Phytophthora capsici in Different Pepper Genotypes.

    Science.gov (United States)

    Silvar, C; Díaz, J; Merino, F

    2005-12-01

    ABSTRACT Reliable and sensitive quantification of Phytophthora capsici in pepper plants is of crucial importance in managing the multiple syndromes caused by this pathogen. A real-time polymerase chain reaction (PCR) assay was developed for the determination of P. capsici in pepper tissues. DNA levels of a highly virulent and a less virulent isolate were measured in different pepper genotypes with varying degrees of resistance. Using SYBR Green and specific primers for P. capsici, the minimal amount of pathogen DNA quantified was 10 pg. Pathogen DNA was recorded as early as 8 h postinoculation. Thereafter, the increase was rapid in susceptible cultivars and slower in resistant ones. The amount of pathogen DNA quantified in each pepper genotype correlated with susceptibility to Phytophthora root rot. Likewise, there was a relationship between the virulence of the pathogen and the degree of colonization. Differences also were found in oomycete amount among pepper tissues, with maximal pathogen biomass occurring in stems. The real-time PCR technique developed in this study was sensitive and robust enough to assess both pathogen development and resistance to Phytophthora root rot in different pepper genotypes.

  14. Insights Into the Etiology of Polerovirus-Induced Pepper Yellows Disease.

    Science.gov (United States)

    Lotos, Leonidas; Olmos, Antonio; Orfanidou, Chrysoula; Efthimiou, Konstantinos; Avgelis, Apostolos; Katis, Nikolaos I; Maliogka, Varvara I

    2017-12-01

    The study of an emerging yellows disease of pepper crops (pepper yellows disease [PYD]) in Greece led to the identification of a polerovirus closely related to Pepper vein yellows virus (PeVYV). Recovery of its full genome sequence by next-generation sequencing of small interfering RNAs allowed its characterization as a new poleroviruses, which was provisionally named Pepper yellows virus (PeYV). Transmission experiments revealed its association with the disease. Sequence similarity and phylogenetic analysis highlighted the common ancestry of the three poleroviruses (PeVYV, PeYV, and Pepper yellow leaf curl virus [PYLCV]) currently reported to be associated with PYD, even though significant genetic differences were identified among them, especially in the C-terminal region of P5 and the 3' noncoding region. Most of the differences observed can be attributed to a modular type of evolution, which produces mosaic-like variants giving rise to these different poleroviruses Overall, similar to other polerovirus-related diseases, PYD is caused by at least three species (PeVYV, PeYV, and PYLCV) belonging to this group of closely related pepper-infecting viruses.

  15. CD8(+) T cells of Listeria monocytogenes-infected mice recognize both linear and spliced proteasome products

    NARCIS (Netherlands)

    Platteel, Anouk C M; Mishto, Michele; Textoris-Taube, Kathrin; Keller, Christin; Liepe, Juliane; Busch, Dirk H; Kloetzel, Peter M; Sijts, Alice J A M

    CD8(+) T cells responding to infection recognize pathogen-derived epitopes presented by MHC class-I molecules. While most of such epitopes are generated by proteasome-mediated antigen cleavage, analysis of tumor antigen processing has revealed that epitopes may also derive from proteasome-catalyzed

  16. Development and evaluation of a sandwich ELISA for quantification of the 20S proteasome in human plasma

    DEFF Research Database (Denmark)

    Dutaud, Dominique; Aubry, Laurent; Henry, Laurent

    2002-01-01

    Because quantification of the 20S proteasome by functional activity measurements is difficult and inaccurate, we have developed an indirect sandwich enzyme-linked immunosorbent assays (ELISA) for quantification of the 20S proteasome in human plasma. This sandwich ELISA uses a combination...

  17. Proteasome inhibitor MG132 sensitizes HPV-positive human cervical cancer cells to rhTRAIL-induced apoptosis

    NARCIS (Netherlands)

    Hougardy, BMT; Maduro, JH; van der Zee, AGJ; de Groot, DJA; van den Heuvel, FAJ; de Vries, EGE; de Jong, S

    2006-01-01

    In cervical carcinogenesis, the p53 tumor suppressor pathway is disrupted by HPV (human papilloma virus) E6 oncogene expression. E6 targets p53 for rapid proteasome-mediated degradation. We therefore investigated whether proteasome inhibition by MG132 could restore wild-type p53 levels and sensitize

  18. Calcitonin Gene-Related Peptide Induces HIV-1 Proteasomal Degradation in Mucosal Langerhans Cells.

    Science.gov (United States)

    Bomsel, Morgane; Ganor, Yonatan

    2017-12-01

    The neuroimmune dialogue between peripheral neurons and Langerhans cells (LCs) within mucosal epithelia protects against incoming pathogens. LCs rapidly internalize human immunodeficiency virus type 1 (HIV-1) upon its sexual transmission and then trans -infect CD4 + T cells. We recently found that the neuropeptide calcitonin gene-related peptide (CGRP), secreted mucosally from peripheral neurons, inhibits LC-mediated HIV-1 trans -infection. In this study, we investigated the mechanism of CGRP-induced inhibition, focusing on HIV-1 degradation in LCs and its interplay with trans -infection. We first show that HIV-1 degradation occurs in endolysosomes in untreated LCs, and functionally blocking such degradation with lysosomotropic agents results in increased trans -infection. We demonstrate that CGRP acts via its cognate receptor and at a viral postentry step to induce faster HIV-1 degradation, but without affecting the kinetics of endolysosomal degradation. We reveal that unexpectedly, CGRP shifts HIV-1 degradation from endolysosomes toward the proteasome, providing the first evidence for functional HIV-1 proteasomal degradation in LCs. Such efficient proteasomal degradation significantly inhibits the first phase of trans -infection, and proteasomal, but not endolysosomal, inhibitors abrogate CGRP-induced inhibition. Together, our results establish that CGRP controls the HIV-1 degradation mode in LCs. The presence of endogenous CGRP within innervated mucosal tissues, especially during the sexual response, to which CGRP contributes, suggests that HIV-1 proteasomal degradation predominates in vivo Hence, proteasomal, rather than endolysosomal, HIV-1 degradation in LCs should be enhanced clinically to effectively restrict HIV-1 trans -infection. IMPORTANCE During sexual transmission, HIV-1 is internalized and degraded in LCs, the resident antigen-presenting cells in mucosal epithelia. Yet during trans -infection, infectious virions escaping degradation are transferred

  19. Computational analysis and modeling of cleavage by the immunoproteasome and the constitutive proteasome

    Directory of Open Access Journals (Sweden)

    Lafuente Esther M

    2010-09-01

    Full Text Available Abstract Background Proteasomes play a central role in the major histocompatibility class I (MHCI antigen processing pathway. They conduct the proteolytic degradation of proteins in the cytosol, generating the C-terminus of CD8 T cell epitopes and MHCI-peptide ligands (P1 residue of cleavage site. There are two types of proteasomes, the constitutive form, expressed in most cell types, and the immunoproteasome, which is constitutively expressed in mature dendritic cells. Protective CD8 T cell epitopes are likely generated by the immunoproteasome and the constitutive proteasome, and here we have modeled and analyzed the cleavage by these two proteases. Results We have modeled the immunoproteasome and proteasome cleavage sites upon two non-overlapping sets of peptides consisting of 553 CD8 T cell epitopes, naturally processed and restricted by human MHCI molecules, and 382 peptides eluted from human MHCI molecules, respectively, using N-grams. Cleavage models were generated considering different epitope and MHCI-eluted fragment lengths and the same number of C-terminal flanking residues. Models were evaluated in 5-fold cross-validation. Judging by the Mathew's Correlation Coefficient (MCC, optimal cleavage models for the proteasome (MCC = 0.43 ± 0.07 and the immunoproteasome (MCC = 0.36 ± 0.06 were obtained from 12-residue peptide fragments. Using an independent dataset consisting of 137 HIV1-specific CD8 T cell epitopes, the immunoproteasome and proteasome cleavage models achieved MCC values of 0.30 and 0.18, respectively, comparatively better than those achieved by related methods. Using ROC analyses, we have also shown that, combined with MHCI-peptide binding predictions, cleavage predictions by the immunoproteasome and proteasome models significantly increase the discovery rate of CD8 T cell epitopes restricted by different MHCI molecules, including A*0201, A*0301, A*2402, B*0702, B*2705. Conclusions We have developed models that are specific

  20. Synergistic apoptosis induction in leukemic cells by the phosphatase inhibitor salubrinal and proteasome inhibitors.

    Directory of Open Access Journals (Sweden)

    Hannes C A Drexler

    Full Text Available Cells adapt to endoplasmic reticulum (ER-stress by arresting global protein synthesis while simultaneously activating specific transcription factors and their downstream targets. These processes are mediated in part by the phosphorylation-dependent inactivation of the translation initiation factor eIF2alpha. Following restoration of homeostasis protein synthesis is resumed when the serine/threonine-protein phosphatase PP1 dephosphorylates and reactivates eIF2alpha. Proteasome inhibitors, used to treat multiple myeloma patients evoke ER-stress and apoptosis by blocking the ER-associated degradation of misfolded proteins (ERAD, however, the role of eIF2alpha phosphorylation in leukemic cells under conditions of proteasome inhibitor-mediated ER stress is currently unclear.Bcr-Abl-positive and negative leukemic cell lines were used to investigate the functional implications of PP1-related phosphatase activities on eIF2alpha phosphorylation in proteasome inhibitor-mediated ER stress and apoptosis. Rather unexpectedly, salubrinal, a recently identified PP1 inhibitor capable to protect against ER stress in various model systems, strongly synergized with proteasome inhibitors to augment apoptotic death of different leukemic cell lines. Salubrinal treatment did not affect the phosphorlyation status of eIF2alpha. Furthermore, the proapoptotic effect of salubrinal occurred independently from the chemical nature of the proteasome inhibitor, was recapitulated by a second unrelated phosphatase inhibitor and was unaffected by overexpression of a dominant negative eIF2alpha S51A variant that can not be phosphorylated. Salubrinal further aggravated ER-stress and proteotoxicity inflicted by the proteasome inhibitors on the leukemic cells since characteristic ER stress responses, such as ATF4 and CHOP synthesis, XBP1 splicing, activation of MAP kinases and eventually apoptosis were efficiently abrogated by the translational inhibitor cycloheximide.Although PP1

  1. Proteasome activity related with the daily physical activity of COPD patients

    Directory of Open Access Journals (Sweden)

    Lee KY

    2017-05-01

    Full Text Available Kang-Yun Lee,1,2,* Tzu-Tao Chen,1,* Ling-Ling Chiang,1,3 Hsiao-Chi Chuang,1,3 Po-Hao Feng,1,2 Wen-Te Liu,1–3 Kuan-Yuan Chen,1 Shu-Chuan Ho1,3 1Division of Pulmonary Medicine, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, New Taipei City, 2Division of Pulmonary Medicine, Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, 3School of Respiratory Therapy, College of Medicine, Taipei Medical University, Taipei, Taiwan *These authors contributed equally to this work Background: COPD is a debilitating disease that affects patients’ daily lives. One’s daily physical activity (DPA decreases due to multifactorial causes, and this decrease is correlated with a poor prognosis in COPD patients. Muscle wasting may at least be partly due to increased activity of the ubiquitin proteasome pathway and apoptosis.Methods: This study investigated the relationships among DPA, circulating proteasome activity, and protein carbonyl in COPD patients and healthy subjects (HSs. This study included 57 participants (42 patients and 15 healthy subjects. Ambulatory DPA was measured using actigraphy, and oxygen saturation was measured with a pulse oximeter.Results: COPD patients had lower DPA, lower 6 min walking distance (6MWD, lower delta saturation pulse oxygenation (SpO2 during the 6MWT, and lower delta SpO2 during DPA than HSs. COPD patients had higher proteasome activity and protein carbonyl than HSs. Circulating proteasome activity was significantly negatively correlated with DPA (r=−0.568, P<0.05 in COPD patients, whereas delta SpO2 during the 6MWT was significantly positively correlated with proteasome activity (r=0.685, P<0.05 in HSs. Protein carbonyl was significantly negatively correlated with the body mass index (r=−0.318, P<0.05, mid-arm circumference (r=0.350, P<0.05, calf circumference (r=0.322, P<0.05, forced expiratory volume in the first second (r=−0.441, P<0

  2. Comparison of primary and secondary 26S rRNA structures in two Tetrahymena species: evidence for a strong evolutionary and structural constraint in expansion segments

    DEFF Research Database (Denmark)

    Engberg, J; Nielsen, Henrik; Lenaers, G

    1990-01-01

    We have determined the nucleotide sequence of the 26S large subunit (LSU) rRNA genes for two Tetrahymena species, T. thermophila and T. pyriformis. The inferred rRNA sequences are presented in their most probable secondary structures based on compensatory mutations, energy, and conservation crite...

  3. FOXOs modulate proteasome activity in human-induced pluripotent stem cells of Huntington's disease and their derived neural cells.

    Science.gov (United States)

    Liu, Yanying; Qiao, Fangfang; Leiferman, Patricia C; Ross, Alan; Schlenker, Evelyn H; Wang, Hongmin

    2017-11-15

    Although it has been speculated that proteasome dysfunction may contribute to the pathogenesis of Huntington's disease (HD), a devastating neurodegenerative disorder, how proteasome activity is regulated in HD affected stem cells and somatic cells remains largely unclear. To better understand the pathogenesis of HD, we analyzed proteasome activity and the expression of FOXO transcription factors in three wild-type (WT) and three HD induced-pluripotent stem cell (iPSC) lines. HD iPSCs exhibited elevated proteasome activity and higher levels of FOXO1 and FOXO4 proteins. Knockdown of FOXO4 but not FOXO1 expression decreased proteasome activity. Following neural differentiation, the HD-iPSC-derived neural progenitor cells (NPCs) demonstrated lower levels of proteasome activity and FOXO expressions than their WT counterparts. More importantly, overexpression of FOXO4 but not FOXO1 in HD NPCs dramatically enhanced proteasome activity. When HD NPCs were further differentiated into DARPP32-positive neurons, these HD neurons were more susceptible to death than WT neurons and formed Htt aggregates under the condition of oxidative stress. Similar to HD NPCs, HD-iPSC-derived neurons showed reduced proteasome activity and diminished FOXO4 expression compared to WT-iPSC-derived neurons. Furthermore, HD iPSCs had lower AKT activities than WT iPSCs, whereas the neurons derived from HD iPSC had higher AKT activities than their WT counterparts. Inhibiting AKT activity increased both FOXO4 level and proteasome activity, indicating a potential role of AKT in regulating FOXO levels. These data suggest that FOXOs modulate proteasome activity, and thus represents a potentially valuable therapeutic target for HD. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  4. Effect of gamma radiation and storage time on the volatile constituents, pirerine, piperettine and sensory quality of pepper

    Energy Technology Data Exchange (ETDEWEB)

    Bahari, I; Ishak, S; Ayub, M K [National Univ. of Malaysia, Bangi, Selangor

    1983-12-01

    The use of gamma radiation in prolonging the storage life of black and white peppers is promising. Doses up to 9 kGy and storage period up to 6 months did not significantly change (P<0.05) the volatile constituents of the peppers. Besides the increase in piperine content of unirradiated pepper there was no change in piperettine and piperine contents of both pepper with respect to increase in dose and storage time. No sensory change was detected for the treatments used (author).

  5. The effect of gamma radiation and storage time on the volatile constituents, pirerine, piperettine and sensory quality of pepper

    International Nuclear Information System (INIS)

    Bahari, I.; Ishak, S.; Ayub, M.K.

    1983-01-01

    The use of gamma radiation in prolonging the storage life of black and white peppers is promising. Doses up to 9 kGy and storage period up to 6 months did not significantly change (P<0.05) the volatile constituents of the peppers. Besides the increase in piperine content of unirradiated pepper there was no change in piperettine and piperine contents of both pepper with respect to increase in dose and storage time. No sensory change was detected for the treatments used (author)

  6. Inhibitory effects of black pepper (Piper nigrum) extracts and compounds on human tumor cell proliferation, cyclooxygenase enzymes, lipid peroxidation and nuclear transcription factor-kappa-B.

    Science.gov (United States)

    Liu, Yunbao; Yadev, Vivek R; Aggarwal, Bharat B; Nair, Muraleedharan G

    2010-08-01

    Black pepper (Piper nigrum) and hot pepper (Capsicum spp.) are widely used in traditional medicines. Although hot Capsicum spp. extracts and its active principles, capsaicinoids, have been linked with anticancer and anti-inflammatory activities, whether black pepper and its active principle exhibit similar activities is not known. In this study, we have evaluated the antioxidant, anti-inflammatory and anticancer activities of extracts and compounds from black pepper by using proinflammatory transcription factor NF-kappaB, COX-1 and -2 enzymes, human tumor cell proliferation and lipid peroxidation (LPO). The capsaicinoids, the alkylamides, isolated from the hot pepper Scotch Bonnet were also used to compare the bioactivities of alkylamides and piperine from black pepper. All compounds derived from black pepper suppressed TNF-induced NF-kappaB activation, but alkyl amides, compound 4 from black pepper and 5 from hot pepper, were most effective. The human cancer cell proliferation inhibitory activities of piperine and alklyl amides in Capsicum and black pepper were dose dependant. The inhibitory concentrations 50% (IC50) of the alklylamides were in the range 13-200 microg/mL. The extracts of black pepper at 200 microg/mL and its compounds at 25 microg/mL inhibited LPO by 45-85%, COX enzymes by 31-80% and cancer cells proliferation by 3.5-86.8%. Overall, these results suggest that black pepper and its constituents like hot pepper, exhibit anti-inflammatory, antioxidant and anticancer activities.

  7. FoxM1 is a general target for proteasome inhibitors.

    Directory of Open Access Journals (Sweden)

    Uppoor G Bhat

    2009-08-01

    Full Text Available Proteasome inhibitors are currently in the clinic or in clinical trials, but the mechanism of their anticancer activity is not completely understood. The oncogenic transcription factor FoxM1 is one of the most overexpressed genes in human tumors, while its expression is usually halted in normal non-proliferating cells. Previously, we established that thiazole antibiotics Siomycin A and thiostrepton inhibit FoxM1 and induce apoptosis in human cancer cells. Here, we report that Siomycin A and thiostrepton stabilize the expression of a variety of proteins, such as p21, Mcl-1, p53 and hdm-2 and also act as proteasome inhibitors in vitro. More importantly, we also found that well-known proteasome inhibitors such as MG115, MG132 and bortezomib inhibit FoxM1 transcriptional activity and FoxM1 expression. In addition, overexpression of FoxM1 specifically protects against bortezomib-, but not doxorubicin-induced apoptosis. These data suggest that negative regulation of FoxM1 by proteasome inhibitors is a general feature of these drugs and it may contribute to their anticancer properties.

  8. Proteasome inhibition by new dual warhead containing peptido vinyl sulfonyl fluorides

    NARCIS (Netherlands)

    Brouwer, Arwin J; Herrero Álvarez, Natalia; Ciaffoni, Adriano; van de Langemheen, Helmus; Liskamp, Rob M J

    2016-01-01

    The success of inhibition of the proteasome by formation of covalent bonds is a major victory over the long held-view that this would lead to binding the wrong targets and undoubtedly lead to toxicity. Great challenges are now found in uncovering ensembles of new moieties capable of forming long

  9. Bioinformatic analysis of functional differences between the immunoproteasome and the constitutive proteasome

    DEFF Research Database (Denmark)

    Kesmir, Can; van Noort, V.; de Boer, R.J.

    2003-01-01

    not yet been quantified how different the specificity of two forms of the proteasome are. The main question, which still lacks direct evidence, is whether the immunoproteasome generates more MHC ligands. Here we use bioinformatics tools to quantify these differences and show that the immunoproteasome...

  10. Modulation of apoptosis sensitivity through the interplay with autophagic and proteasomal degradation pathways

    Science.gov (United States)

    Delgado, M E; Dyck, L; Laussmann, M A; Rehm, M

    2014-01-01

    Autophagic and proteasomal degradation constitute the major cellular proteolysis pathways. Their physiological and pathophysiological adaptation and perturbation modulates the relative abundance of apoptosis-transducing proteins and thereby can positively or negatively adjust cell death susceptibility. In addition to balancing protein expression amounts, components of the autophagic and proteasomal degradation machineries directly interact with and co-regulate apoptosis signal transduction. The influence of autophagic and proteasomal activity on apoptosis susceptibility is now rapidly gaining more attention as a significant modulator of cell death signalling in the context of human health and disease. Here we present a concise and critical overview of the latest knowledge on the molecular interplay between apoptosis signalling, autophagy and proteasomal protein degradation. We highlight that these three pathways constitute an intricate signalling triangle that can govern and modulate cell fate decisions between death and survival. Owing to rapid research progress in recent years, it is now possible to provide detailed insight into the mechanisms of pathway crosstalk, common signalling nodes and the role of multi-functional proteins in co-regulating both protein degradation and cell death. PMID:24457955

  11. Role of Proteasome-Dependent Protein Degradation in Long-Term Operant Memory in "Aplysia"

    Science.gov (United States)

    Lyons, Lisa C.; Gardner, Jacob S.; Gandour, Catherine E.; Krishnan, Harini C.

    2017-01-01

    We investigated the in vivo role of protein degradation during intermediate (ITM) and long-term memory (LTM) in "Aplysia" using an operant learning paradigm. The proteasome inhibitor MG-132 inhibited the induction and molecular consolidation of LTM with no effect on ITM. Remarkably, maintenance of steady-state protein levels through…

  12. Amyloid-β secretion, generation, and lysosomal sequestration in response to proteasome inhibition

    DEFF Research Database (Denmark)

    Agholme, Lotta; Hallbeck, Martin; Benedikz, Eirikur

    2012-01-01

    , as the autophagosome has been suggested as a site of amyloid-β (Aβ) generation. In this study, we investigated the effect of proteasome inhibition on Aβ accumulation and secretion, as well as the processing of amyloid-β protein precursor (AβPP) in AβPP(Swe) transfected SH-SY5Y neuroblastoma cells. We show...

  13. BAG3-dependent noncanonical autophagy induced by proteasome inhibition in HepG2 cells.

    Science.gov (United States)

    Liu, Bao-Qin; Du, Zhen-Xian; Zong, Zhi-Hong; Li, Chao; Li, Ning; Zhang, Qiang; Kong, De-Hui; Wang, Hua-Qin

    2013-06-01

    Emerging lines of evidence have shown that blockade of ubiquitin-proteasome system (UPS) activates autophagy. The molecular players that regulate the relationship between them remain to be elucidated. Bcl-2 associated athanogene 3 (BAG3) is a member of the BAG co-chaperone family that regulates the ATPase activity of heat shock protein 70 (HSP70) chaperone family. Studies on BAG3 have demonstrated that it plays multiple roles in physiological and pathological processes, including antiapoptotic activity, signal transduction, regulatory role in virus infection, cell adhesion and migration. Recent studies have attracted much attention on its role in initiation of autophagy. The current study, for the first time, demonstrates that proteasome inhibitors elicit noncanonical autophagy, which was not suppressed by inhibitors of class III phosphatidylinositol 3-kinase (PtdIns3K) or shRNA against Beclin 1 (BECN1). In addition, we demonstrate that BAG3 is ascribed to activation of autophagy elicited by proteasome inhibitors and MAPK8/9/10 (also known as JNK1/2/3 respectively) activation is also implicated via upregulation of BAG3. Moreover, we found that noncanonical autophagy mediated by BAG3 suppresses responsiveness of HepG2 cells to proteasome inhibitors.

  14. Combination of Proteasomal Inhibitors Lactacystin and MG132 Induced Synergistic Apoptosis in Prostate Cancer Cells

    Directory of Open Access Journals (Sweden)

    Robert B. Shirley

    2005-12-01

    Full Text Available The proteasome inhibitor Velcade (bortezomib/PS-341 has been shown to block the targeted proteolytic degradation of short-lived proteins that are involved in cell maintenance, growth, division, and death, advocating the use of proteasomal inhibitors as therapeutic agents. Although many studies focused on the use of one proteasomal inhibitor for therapy, we hypothesized that the combination of proteasome inhibitors Lactacystin (AG Scientific, Inc., San Diego, CA and MG132 (Biomol International, Plymouth Meeting, PA may be more effective in inducing apoptosis. Additionally, this regimen would enable the use of sublethal doses of individual drugs, thus reducing adverse effects. Results indicate a significant increase in apoptosis when LNCaP prostate cancer cells were treated with increasing levels of Lactacystin, MG132, or a combination of sublethal doses of these two inhibitors. Furthermore, induction in apoptosis coincided with a significant loss of IKKα, IKKβ, and IKKγ proteins and NFκB activity. In addition to describing effective therapeutic agents, we provide a model system to facilitate the investigation of the mechanism of action of these drugs and their effects on the IKK-NFκB axis.

  15. Application of microwaves for microbial load reduction in black pepper (Piper nigrum L.).

    Science.gov (United States)

    Jeevitha, G Chengaiyan; Sowbhagya, H Bogegowda; Hebbar, H Umesh

    2016-09-01

    Black pepper (Piper nigrum L.) is exposed to microbial contamination which could potentially create public health risk and also rejection of consignments in the export market due to non-adherance to microbial safety standards. The present study investigates the use of microwave (MW) radiation for microbial load reduction in black pepper and analyses the effect on quality. Black pepper was exposed to MWs at two different power levels (663 and 800 W) at an intensity of 40 W g(-1) for different time intervals (1-15 min) and moisture content (110 and 260 g kg(-1) on a wet basis). The exposure of black pepper to MWs at 663 W for 12.5 min was found to be sufficient to reduce the microbial load to the permissible level suggested by the International Commission on Microbiological Specifications for Foods and the European Spice Association. The retention of volatile oil, piperine and resin was 91.3 ± 0.03, 87.6 ± 0.02 and 90.7 ± 0.05%, respectively, in MW-treated black pepper. The final moisture content after MW treatment was found to be 100 ± 1 g kg(-1) for black pepper containing initial moisture of 260 ± 3 g kg(-1) . These results suggest that MW heating can be effectively used for microbial load reduction of black pepper without a significant loss in product quality. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  16. Molecular detection and characterization of a phytoplasma from xianlajiao chili pepper in shaanxi province, china

    International Nuclear Information System (INIS)

    Guo, C.M.; Lu, M.H.; Zhao, Z.L.; Hu, J.C.; Nie, W.Y.; Guo, J.W.

    2017-01-01

    Surveys were performed in the main Xianlajiao chili pepper-producing areas of Shaanxi Province to determine whether a phytoplasma was associated with the incidence of Arbuscular Lobular Disease (ALD). During July 2012 and 2013, 92 and 86 chili pepper plants, respectively, showing ALD-like symptoms were collected from 12 counties and the cities of Baoji, Weinan, Xianyang, and Xian. Samples from paulownia trees and plantain with symptoms of paulownia witches' broom and little leaf, respectively, located in pepper fields in Baoji city, Fengxiang county were also collected. Universal DNA primers for amplification of the phytoplasma 16S rRNA gene were used for nested PCR assays and DNA sequencing. Phytoplasma DNA was amplified from two chili pepper plants, the paulownia trees, and the plantain collected from Baoji city. The phylogenetic analysis of 16S rDNA sequences of the phytoplasmas detected in the chili pepper confirmed that these phytoplasmas are members of the 16SrI group "Candidatus Phytoplasma asteris". The results of virtual RFLP analysis and sequence homology comparison showed that the two Xianlajiao phytoplasmas have high homology with phytoplasmas 16SrI-B and 16SrI-M. Transmission electron microscopy assays revealed the presence of typical phytoplasma pleomorphic bodies in the phloem of phytoplasma-infected Xianlajiao chili pepper plants. Although our results did not establish a strong association between the presence of the phytoplasma and ALD, the simultaneous detection of a 16SrI phytoplasma in two symptomatic Xianlajiao chili peppers, the paulownia trees, and the plantain suggests a complex epidemiology related to the 16SrI phytoplasma pathogen in these new plant hosts. (author)

  17. Management of blight of bell pepper (Capsicum annuum var. grossum) caused by Drechslera bicolor.

    Science.gov (United States)

    Jadon, Kuldeep Singh; Shah, Rakesh; Gour, Hari Narayan; Sharma, Pankaj

    Sweet or bell pepper is a member of the Solanaceae family and is regarded as one of the most popular and nutritious vegetable. Blight, in the form of leaf and fruit blight, has been observed to infect bell pepper crops cultivated at the horticulture farm in Rajasthan College of Agriculture, Udaipur, India. Based on disease severity, we attempted to curb this newly emerged problem using different fungicides, plant extracts, bio-control agents, and commercial botanicals against the fungus in laboratory and pot experiments. Bio-control agent Trichoderma viride and plant growth promoting Rhizobacteria (PGPR) isolate Neist-2 were found to be quite effective against bell pepper blight. All evaluated fungicides, botanicals, commercial botanicals, and bio-control agents in vitro were further studied as seed dressers and two foliar sprays at ten days interval in pot experiments. The combinations of Vitavax, PGPR isolate Neist-2, and Mehandi extract were found to be very effective against bell pepper blight followed by Vitavax, T. viride, and Mehandi extract used individually. All treatments in the pot experiments were found to significantly reduce seedling mortality and enhance plant biomass of bell pepper. Thus, these experimental findings suggest that a better integrated management of bell pepper blight could be achieved by conducting field trials in major bell pepper- and chilli-cultivated areas of the state. Besides fungicides, different botanicals and commercial botanicals also seem to be promising treatment options. Therefore, the outcome of the present study provides an alternate option of fungicide use in minimizing loss caused by Drechslera bicolor. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  18. Azithromycin attenuates myofibroblast differentiation and lung fibrosis development through proteasomal degradation of NOX4.

    Science.gov (United States)

    Tsubouchi, Kazuya; Araya, Jun; Minagawa, Shunsuke; Hara, Hiromichi; Ichikawa, Akihiro; Saito, Nayuta; Kadota, Tsukasa; Sato, Nahoko; Yoshida, Masahiro; Kurita, Yusuke; Kobayashi, Kenji; Ito, Saburo; Fujita, Yu; Utsumi, Hirofumi; Yanagisawa, Haruhiko; Hashimoto, Mitsuo; Wakui, Hiroshi; Yoshii, Yutaka; Ishikawa, Takeo; Numata, Takanori; Kaneko, Yumi; Asano, Hisatoshi; Yamashita, Makoto; Odaka, Makoto; Morikawa, Toshiaki; Nakayama, Katsutoshi; Nakanishi, Yoichi; Kuwano, Kazuyoshi

    2017-08-03

    Accumulation of profibrotic myofibroblasts is involved in the process of fibrosis development during idiopathic pulmonary fibrosis (IPF) pathogenesis. TGFB (transforming growth factor β) is one of the major profibrotic cytokines for myofibroblast differentiation and NOX4 (NADPH oxidase 4) has an essential role in TGFB-mediated cell signaling. Azithromycin (AZM), a second-generation antibacterial macrolide, has a pleiotropic effect on cellular processes including proteostasis. Hence, we hypothesized that AZM may regulate NOX4 levels by modulating proteostasis machineries, resulting in inhibition of TGFB-associated lung fibrosis development. Human lung fibroblasts (LF) were used to evaluate TGFB-induced myofibroblast differentiation. With respect to NOX4 regulation via proteostasis, assays for macroautophagy/autophagy, the unfolded protein response (UPR), and proteasome activity were performed. The potential anti-fibrotic property of AZM was examined by using bleomycin (BLM)-induced lung fibrosis mouse models. TGFB-induced NOX4 and myofibroblast differentiation were clearly inhibited by AZM treatment in LF. AZM-mediated NOX4 reduction was restored by treatment with MG132, a proteasome inhibitor. AZM inhibited autophagy and enhanced the UPR. Autophagy inhibition by AZM was linked to ubiquitination of NOX4 via increased protein levels of STUB1 (STIP1 homology and U-box containing protein 1), an E3 ubiquitin ligase. An increased UPR by AZM was associated with enhanced proteasome activity. AZM suppressed lung fibrosis development induced by BLM with concomitantly reduced NOX4 protein levels and enhanced proteasome activation. These results suggest that AZM suppresses NOX4 by promoting proteasomal degradation, resulting in inhibition of TGFB-induced myofibroblast differentiation and lung fibrosis development. AZM may be a candidate for the treatment of the fibrotic lung disease IPF.

  19. Inhibition of autophagy induced by proteasome inhibition increases cell death in human SHG-44 glioma cells.

    Science.gov (United States)

    Ge, Peng-Fei; Zhang, Ji-Zhou; Wang, Xiao-Fei; Meng, Fan-Kai; Li, Wen-Chen; Luan, Yong-Xin; Ling, Feng; Luo, Yi-Nan

    2009-07-01

    The ubiquitin-proteasome system (UPS) and lysosome-dependent macroautophagy (autophagy) are two major intracellular pathways for protein degradation. Recent studies suggest that proteasome inhibitors may reduce tumor growth and activate autophagy. Due to the dual roles of autophagy in tumor cell survival and death, the effect of autophagy on the destiny of glioma cells remains unclear. In this study, we sought to investigate whether inhibition of the proteasome can induce autophagy and the effects of autophagy on the fate of human SHG-44 glioma cells. The proteasome inhibitor MG-132 was used to induce autophagy in SHG-44 glioma cells, and the effect of autophagy on the survival of SHG-44 glioma cells was investigated using an autophagy inhibitor 3-MA. Cell viability was measured by MTT assay. Apoptosis and cell cycle were detected by flow cytometry. The expression of autophagy related proteins was determined by Western blot. MG-132 inhibited cell proliferation, induced cell death and cell cycle arrest at G(2)/M phase, and activated autophagy in SHG-44 glioma cells. The expression of autophagy-related Beclin-1 and LC3-I was significantly up-regulated and part of LC3-I was converted into LC3-II. However, when SHG-44 glioma cells were co-treated with MG-132 and 3-MA, the cells became less viable, but cell death and cell numbers at G(2)/M phase increased. Moreover, the accumulation of acidic vesicular organelles was decreased, the expression of Beclin-1 and LC3 was significantly down-regulated and the conversion of LC3-II from LC3-I was also inhibited. Inhibition of the proteasome can induce autophagy in human SHG-44 glioma cells, and inhibition of autophagy increases cell death. This discovery may shed new light on the effect of autophagy on modulating the fate of SHG-44 glioma cells.Acta Pharmacologica Sinica (2009) 30: 1046-1052; doi: 10.1038/aps.2009.71.

  20. A cytosolic protein factor from the naked mole-rat activates proteasomes of other species and protects these from inhibition

    Science.gov (United States)

    Rodriguez, Karl A.; Osmulski, Pawel A.; Pierce, Anson; Weintraub, Susan T.; Gaczynska, Maria; Buffenstein, Rochelle

    2015-01-01

    The naked mole-rat maintains robust proteostasis and high levels of proteasome-mediated proteolysis for most of its exceptional (~31y) life span. Here, we report that the highly active proteasome from the naked mole-rat liver resists attenuation by a diverse suite of proteasome-specific small molecule inhibitors. Moreover, mouse, human, and yeast proteasomes exposed to the proteasome-depleted, naked mole-rat cytosolic fractions, recapitulate the observed inhibition resistance, and mammalian proteasomes also show increased activity. Gel filtration coupled with mass spectrometry and atomic force microscopy indicates that these traits are supported by a protein factor that resides in the cytosol. This factor interacts with the proteasome and modulates its activity. Although HSP72 and HSP40 (Hdj1) are among the constituents of this factor, the observed phenomenon, such as increasing peptidase activity and protecting against inhibition cannot be reconciled with any known chaperone functions. This novel function may contribute to the exceptional protein homeostasis in the naked mole-rat and allow it to successfully defy aging. PMID:25018089

  1. Management of chili pepper root rot and wilt (caused by Phytophthora nicotianae) by grafting onto resistant rootstock

    OpenAIRE

    Mourad SAADOUN; Mohamed Bechir ALLAGUI

    2013-01-01

    Root rot and plant wilting caused by Phytophthora nicotianae is a severe disease of chili pepper (Capsicum annuum L.) in open fields and under greenhouse production in Tunisia. Chili pepper grafting for disease manage- ment is attracting increased interest in recent years. Using the tube grafting technique, different compatible scion/rootstock combinations were obtained with the wild-type pepper SCM334 and the local chili pepper cultivars ‘Beldi’ and ‘Baker’. SCM334 was resistant to P. nicoti...

  2. A study on the comparison of antioxidant effects between hot pepper extract and capsaicin

    Directory of Open Access Journals (Sweden)

    Hyeon Min Gang

    2008-03-01

    Full Text Available Objective : The objective of this study was to compare the antioxidant effects of hot pepper extract and capsaicin. Methods : In vitro, antioxidant activities were examined by DPPH radical scavenging activity, total antioxidant capacity(TAC, oxygen radical scavenging capacity(ORAC, inhibition of induced lipid peroxidation using liver mitochonria and total phenolic contents. Results : 1. DPPH free radical scavenging actiivities at the concentrations of both 1 and 10㎎/㎖ were 1.2 to 1.9 times higher in capsaicin than in hot pepper extract. The concentration of capsaicin required for 50% radical scavenging was lower than that of hot pepper extract(3.9 vs 5.9㎎/㎖, indicating that capsaicin had higher DPPH radical scavenging activity than hot pepper extract. 2. Total antioxidant capacities of capsaicin at the concentrations of 0.1 and 1mg/ml(13.8 and 41.3 nmol Trolox equivalent were not significantly different from those at the concentrations of 1 and 10㎎/㎖(11.4 and 41.2nmol Trolox equivalent, indicating that capsaicin showed 10 times higher ABTS radical scavenging activity compared to hot pepper extract. 3. ORAC of capsaicin at the concentrations of 1, 5, 10 and 100 mg/ml were 0.04, 0.17, 0.29 and 1.74nmol gallic acid equivalent, respectively. On the other hand, ORAC of hot pepper extract at the concentrations of 1, 5, 10 and 100㎍/㎖ were 0.15, 0.44, 0.75 and 2.49nmol gallic acid equivalent, respectively, indicating that capsaicin showed higher peroxyl radical scavenging activity than hot pepper extract. 4. Inhibition of lipid peroxidation caused by hot pepper extract at the concentrations of 1 and 10mg/㎖ were 12.2 and 61.4%, respectively. Inhibition of lipid peroxidation caused by capsaicin at the concentrations of 1 and 10㎎/㎖l were 64.0 and 96.8%, respectively. Thus capsaicin showed 10 times stronger effect in inhibiton of lipid peroxidation than hot pepper extract. 5. Total phenolic contents of hot pepper extract at the

  3. Structural characterization of the bacterial proteasome homolog BPH reveals a tetradecameric double-ring complex with unique inner cavity properties.

    Science.gov (United States)

    Fuchs, Adrian C D; Maldoner, Lorena; Hipp, Katharina; Hartmann, Marcus D; Martin, Jörg

    2018-01-19

    Eukaryotic and archaeal proteasomes are paradigms for self-compartmentalizing proteases. To a large extent, their function requires interplay with hexameric ATPases associated with diverse cellular activities (AAA+) that act as substrate unfoldases. Bacteria have various types of self-compartmentalizing proteases; in addition to the proteasome itself, these include the proteasome homolog HslV, which functions together with the AAA+ HslU; the ClpP protease with its partner AAA+ ClpX; and Anbu, a recently characterized ancestral proteasome variant. Previous bioinformatic analysis has revealed a novel bacterial member of the proteasome family Betaproteobacteria proteasome homolog (BPH). Using cluster analysis, we here affirmed that BPH evolutionarily descends from HslV. Crystal structures of the Thiobacillus denitrificans and Cupriavidus metallidurans BPHs disclosed a homo-oligomeric double-ring architecture in which the active sites face the interior of the cylinder. Using small-angle X-ray scattering (SAXS) and electron microscopy averaging, we found that BPH forms tetradecamers in solution, unlike the dodecamers seen in HslV. Although the highly acidic inner surface of BPH was in striking contrast to the cavity characteristics of the proteasome and HslV, a classical proteasomal reaction mechanism could be inferred from the covalent binding of the proteasome-specific inhibitor epoxomicin to BPH. A ligand-bound structure implied that the elongated BPH inner pore loop may be involved in substrate recognition. The apparent lack of a partner unfoldase and other unique features, such as Ser replacing Thr as the catalytic residue in certain BPH subfamilies, suggest a proteolytic function for BPH distinct from those of known bacterial self-compartmentalizing proteases. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Attachment of 13 Types of Foodborne Bacteria to Jalapeño and Serrano Peppers and Antibacterial Effect of Roselle Calyx Extracts, Sodium Hypochlorite, Colloidal Silver, and Acetic Acid against These Foodborne Bacteria on Peppers.

    Science.gov (United States)

    Rangel-Vargas, Esmeralda; Gómez-Aldapa, Carlos A; Falfan-Cortes, Reyna N; Rodríguez-Marín, María L; Godínez-Oviedo, Angélica; Acevedo-Sandoval, Otilio A; Castro-Rosas, Javier

    2017-03-01

    Chili peppers are a very important crop in Mexico. However, these peppers have been associated with Salmonella infection outbreaks in the United States, and Salmonella and diarrheagenic Escherichia coli pathotypes have been isolated from jalapeño and serrano peppers in Mexico. To decrease microbial contamination of fruits and vegetables, chemical agents are commonly used; however, chemical agents used to eliminate pathogenic bacteria on vegetables have a limited antimicrobial effect. Roselle ( Hibiscus sabdariffa ) calyces have been reported to have an antimicrobial effect on pathogenic bacteria. In the present study, the antibacterial effect of four roselle calyx extracts (water, methanol, acetone, and ethyl acetate), sodium hypochlorite, colloidal silver, and acetic acid against foodborne bacteria was evaluated on contaminated jalapeño and serrano peppers. The 13 types of foodborne bacteria evaluated were Listeria monocytogenes , Shigella flexneri , Salmonella Typhimurium, Salmonella Typhi, Salmonella Montevideo, Staphylococcus aureus , E. coli O157:H7, five E. coli pathotypes (Shiga toxin producing, enteropathogenic, enterotoxigenic, enteroinvasive, and enteroaggregative), and Vibrio cholerae O1. All 13 types attached to both pepper types, with no significant differences in attachment between jalapeño and serrano peppers. Roselle calyx extract treatment resulted in a greater reduction in levels of all foodborne bacteria than did treatment with sodium hypochlorite, colloidal silver, and acetic acid on both pepper types. Roselle calyx extracts may be a useful for disinfection of chili peppers in the field, processing plants, restaurants, and homes.

  5. Proteomic analysis reveals strong mitochondrial involvement in cytoplasmic male sterility of pepper (Capsicum annuum L.).

    Science.gov (United States)

    Guo, Jinju; Wang, Peng; Cheng, Qing; Sun, Limin; Wang, Hongyu; Wang, Yutong; Kao, Lina; Li, Yanan; Qiu, Tuoyu; Yang, Wencai; Shen, Huolin

    2017-09-25

    Although cytoplasmic male sterility (CMS) is widely used for developing pepper hybrids, its molecular mechanism remains unclear. In this study, we used a high-throughput proteomics method called label-free to compare protein abundance across a pepper CMS line (A-line) and its isogenic maintainer line (B-line). Data are available via ProteomeXchange with identifier PXD006104. Approximately 324 differentially abundant protein species were identified and quantified; among which, 47 were up-accumulated and 140 were down-accumulated in the A-line; additionally, 75 and 62 protein species were specifically accumulated in the A-line and B-line, respectively. Protein species involved in pollen exine formation, pyruvate metabolic processes, the tricarboxylic acid cycle, the mitochondrial electron transport chain, and oxidative stress response were observed to be differentially accumulated between A-line and B-line, suggesting their potential roles in the regulation of pepper pollen abortion. Based on our data, we proposed a potential regulatory network for pepper CMS that unifies these processes. Artificial emasculation is a major obstacle in pepper hybrid breeding for its high labor cost and poor seed purity. While the use of cytoplasmic male sterility (CMS) in hybrid system is seriously frustrated because a long time is needed to cultivate male sterility line and its isogenic restore line. Transgenic technology is an effective and rapid method to obtain male sterility lines and its widely application has very important significance in speeding up breeding process in pepper. Although numerous studies have been conducted to select the genes related to male sterility, the molecular mechanism of cytoplasmic male sterility in pepper remains unknown. In this study, we used the high-throughput proteomic method called "label-free", coupled with liquid chromatography-quadrupole mass spectrometry (LC-MS/MS), to perform a novel comparison of expression profiles in a CMS pepper line

  6. Anatomical traits of sweet pepper (Capsicum annuum L. fruit

    Directory of Open Access Journals (Sweden)

    Elżbieta Weryszko-Chmielewska

    2012-12-01

    Full Text Available The micromorphology of the epidermis as well as the anatomy of the pericarp and fruit pedicle in Capsicum annuum L., cv. 'Red Knight F1', were studied using light and scanning electron microscopy. The pericarp was found to consist of an epidermis with strongly thickened outer walls, several layers of tangential and angular collenchyma as well as multi-layered parenchyma composed of cells of varying size in which very large lobed nuclei were observed. Chromoplasts were found in the cells of the above-mentioned tissues. The inner epidermis of the pericarp was characterized by thick cell walls and numerous straight pits. Among the tissues of the fruit pedicle, we observed epidermis with numerous stomata, collenchyma, chlorenchyma with very large intercellular spaces, small clusters of sclerenchyma, secondary phloem and xylem as well live and dead cells of the pith which were characterized by the presence of thin walls with numerous pits. The structural traits of the pericarp of the red pepper cultivar under study show adaptations to significantly reduced transpiration, which is an important feature during storage. At the same time, the strongly thickened and cutinized walls of the fruit contribute to a reduction in its digestibility and impede nutrient penetration in non-root feeding.

  7. Proteome analysis of bell pepper (Capsicum annuum L.) chromoplasts.

    Science.gov (United States)

    Siddique, Muhammad Asim; Grossmann, Jonas; Gruissem, Wilhelm; Baginsky, Sacha

    2006-12-01

    We report a comprehensive proteome analysis of chromoplasts from bell pepper (Capsicum annuum L.). The combination of a novel strategy for database-independent detection of proteins from tandem mass spectrometry (MS/MS) data with standard database searches allowed us to identify 151 proteins with a high level of confidence. These include several well-known plastid proteins but also novel proteins that were not previously reported from other plastid proteome studies. The majority of the identified proteins are active in plastid carbohydrate and amino acid metabolism. Among the most abundant individual proteins are capsanthin/capsorubin synthase and fibrillin, which are involved in the synthesis and storage of carotenoids that accumulate to high levels in chromoplasts. The relative abundances of the identified chromoplast proteins differ remarkably compared with their abundances in other plastid types, suggesting a chromoplast-specific metabolic network. Our results provide an overview of the major metabolic pathways active in chromoplasts and extend existing knowledge about prevalent metabolic activities of different plastid types.

  8. Development of virulence to Meloidogyne incognita on resistant pepper rootstocks

    Energy Technology Data Exchange (ETDEWEB)

    Ros-Ibanez, C.; Robertson, L.; Martinez-Lluch, M. C.; Cano-Garcia, A.; Lacasa-Plasencia, A.

    2014-06-01

    The root-knot nematode (RKN) Meloidogyne incognita is a major soil parasite of pepper crops in greenhouses in Southeast Spain. Due to the limitations of the use of soil fumigants, grafting plants on resistant rootstocks (R-rootstocks) has become an important alternative to chemical nematicides. The repeated use of R-rootstocks can bring about the selection of virulent populations capable of overcoming resistance. We carried out a six-year investigation on resistant rootstocks in a naturally M. incognita infested greenhouse, and found that two successive years of growing plants grafted on R-rootstocks Atlante (ATL) were sufficient to overcome resistance (galling index 1.5 and 5.6 in the first and second years respectively). A large variability was observed between several R-rootstocks. Two R-rootstocks (C19 and Snooker) behaved like ATL while two others (Terrano and DRO 8801) were not infected by RKN. Laboratory studies with the same R-rootstocks, inoculated with two nematode isolates (avirulent and virulent against ATL) confirmed the greenhouse results, indicating that some rootstocks may be infested by virulent populations and others may not. It suggests that different R-genes, which are differentially overcome by RKN, have been introgressed into the rootstocks. This may have consequences for the management of resistant rootstocks in the field. (Author)

  9. Phytotoxicity of sarmentine isolated from long pepper (Piper longum) fruit.

    Science.gov (United States)

    Huang, Huazhang; Morgan, Christy M; Asolkar, Ratnakar N; Koivunen, Marja E; Marrone, Pamela G

    2010-09-22

    Discovery of novel natural herbicides has become crucial to overcome increasing weed resistance and environmental issues. In this article, we describe the finding that a methanol extract of dry long pepper (Piper longum L.) fruits is phytotoxic to lettuce (Lactuca sativa L.) seedlings. The bioassay-guided fractionation and purification of the crude extract led to isolation of sarmentine (1), a known compound, as the active principle. Phytotoxicity of 1 was examined with a variety of seedlings of field crops and weeds. Results indicated that 1 was a contact herbicide and possessed broad-spectrum herbicidal activity. Moreover, a series of sarmentine analogues were then synthesized to study the structure-activity relationship (SAR). SAR studies suggested that phytotoxicity of sarmentine and its analogues was specific due to chemical structures, i.e., the analogues of the acid moiety of 1 were active, but the amine and its analogues were inactive; the ester analogues and amide analogues with a primary amine of 1 were also inactive. In addition, quantification of 1 from different resources of the dry P. longum fruits using liquid chromatography-mass spectrometry showed a wide variation, ranging from almost zero to 0.57%. This study suggests that 1 has potential as an active lead molecule for synthesized herbicides as well as for bioherbicides derived from natural resources.

  10. IMPROVEMENT OF SHELF LIFE QUALITY OF GREEN BELL PEPPERS USING EDIBLE COATING FORMULATIONS

    Directory of Open Access Journals (Sweden)

    Emilio Ochoa-Reyes

    2013-06-01

    Full Text Available In Latin-America, there are countries with high production levels of green bell peppers, which requires of new strategies of conservation for their international trade. Traditional techniques of preservations do not guarantee to prolong the shelf life of these kinds of fruits, for this reason, in the present study, the Influence of different edible coating formulations on shelf-life quality of green bell peppers was studied. Three different biopolymers (pectin, arabic, and xanthan gums were evaluated in mixtures with candelilla wax as hydrophobic phase, jojoba oil as plasticizer and a crude extract of polyphenols as source of bioactive compounds. Green bell peppers were immersion-treated and then stored at room temperature. Response variables were: weight loss, color, appearance, pH, total soluble solids and firmness changes which were kinetically determined. All peppers treated with edible-coating showed a significant difference (Tukey, p≤0.05 in weight loss compared to control treatment (without edible coating, while a lower level of deterioration was observed in fruits treated with edible coating formulated with arabic gum, but appearance remained similar among fruits treated with different edible coatings. Use of mixtures of biopolymers, candelilla wax, jojoba oil and polyphenols to develop edible and functionalized coatings significantly extended shelf life of green bell pepper.

  11. Biocontrol of Phytophthora Blight and Anthracnose in Pepper by Sequentially Selected Antagonistic Rhizobacteria against Phytophthora capsici.

    Science.gov (United States)

    Sang, Mee Kyung; Shrestha, Anupama; Kim, Du-Yeon; Park, Kyungseok; Pak, Chun Ho; Kim, Ki Deok

    2013-06-01

    We previously developed a sequential screening procedure to select antagonistic bacterial strains against Phytophthora capsici in pepper plants. In this study, we used a modified screening procedure to select effective biocontrol strains against P. capsici; we evaluated the effect of selected strains on Phytophthora blight and anthracnose occurrence and fruit yield in pepper plants under field and plastic house conditions from 2007 to 2009. We selected four potential biocontrol strains (Pseudomonas otitidis YJR27, P. putida YJR92, Tsukamurella tyrosinosolvens YJR102, and Novosphingobium capsulatum YJR107) among 239 bacterial strains. In the 3-year field tests, all the selected strains significantly (P anthracnose incidence in at least one of the test years, but their biocontrol activities were variable. In addition, strains YJR27, YJR92, and YJR102, in certain harvests, increased pepper fruit numbers in field tests and red fruit weights in plastic house tests. Taken together, these results indicate that the screening procedure is rapid and reliable for the selection of potential biocontrol strains against P. capsici in pepper plants. In addition, these selected strains exhibited biocontrol activities against anthracnose, and some of the strains showed plant growth-promotion activities on pepper fruit.

  12. Fire feedbacks facilitate invasion of pine savannas by Brazilian pepper (Schinus terebinthifolius).

    Science.gov (United States)

    Stevens, Jens T; Beckage, Brian

    2009-10-01

    * Fire disturbance can mediate the invasion of ecological communities by nonnative species. Nonnative plants that modify existing fire regimes may initiate a positive feedback that can facilitate their continued invasion. Fire-sensitive plants may successfully invade pyrogenic landscapes if they can inhibit fire in the landscape. * Here, we investigated whether the invasive shrub Brazilian pepper (Schinus terebinthifolius) can initiate a fire-suppression feedback in a fire-dependent pine savanna ecosystem in the southeastern USA. * We found that prescribed burns caused significant (30-45%) mortality of Brazilian pepper at low densities and that savannas with more frequent fires contained less Brazilian pepper. However, high densities of Brazilian pepper reduced fire temperature by up to 200 degrees C, and experienced as much as 80% lower mortality. * A cellular automaton model was used to demonstrate that frequent fire may control low-density populations, but that Brazilian pepper may reach a sufficient density during fire-free periods to initiate a positive feedback that reduces the frequency of fire and converts the savanna to an invasive-dominated forest.

  13. The effect of irradiation in the preservation of pink pepper (Schinus terebinthifolius Raddi)

    International Nuclear Information System (INIS)

    Régia Marques de Souza, Adriana; Arthur, Valter; Pires Nogueira, Danielle

    2012-01-01

    Pink peppers, also known as “pimenta-rosa” and “poivre rose”, are the fruit of Schinus terebinthifolius Raddi, a species of pepper cultivated in Brazil, and have great potential for the exploration of uses. In efforts to lengthen the shelf life of this pepper, the purpose of this study was to evaluate the effect of different doses of radiation on its physical composition and color. The pink pepper samples were irradiated with doses of 0, 0.2, 0.4, 0.8 and 1.6 kGy, and the moisture, ash and lipid contents, pH and color were analyzed. The moisture content, lipid content and pH analysis indicated effects due to the irradiation (p>0.05) in which the higher doses resulted in decreases in the attribute. In contrast, there were no significant differences for the ash analysis (p ⁎ and b ⁎ were the most affected by the intermediate doses (0.2 and 0.8 kGy), which induced their elevation, enhancing the reddish and yellowish colors. Based on the presented data, irradiation is as an alternative preservation process for pink peppers.

  14. Honeybees as an aid in improving labour conditions in sweet bell pepper greenhouses: reduction of pollen allergy

    NARCIS (Netherlands)

    Steen, van der J.J.M.; Blacquière, T.; Jong, de N.W.; Groot, de H.

    2004-01-01

    Sweet bell pepper is the most important greenhouse vegetable crop in the Netherlands. It is grown on an area of 10,000 hectares, and about 8000 people are working in these greenhouses. One third of these workers sooner or later develop an occupational allergy to the sweet bell pepper pollen. The

  15. Development of a sweet cherry pepper line with resistance to the southern root-knot nematode Meloidogyne incognita

    Science.gov (United States)

    The southern root-knot nematode (Meloidogyne incognita) is a major pathogen of pepper (Capsicum spp.), causing significant yield losses in heavily infected plants. The N-gene confers resistance to M. incognita, and has been successfully used to mitigate nematode damage in specific pepper varieties f...

  16. Potential for breeding sweet pepper adapted to cooler growing conditions : a physiological and genetic analysis of growth traits in Capsicum

    NARCIS (Netherlands)

    Swart, de E.A.M.

    2007-01-01

    Sweet pepper (Capsicum annuum L.) is an important greenhouse crop in theNetherlands. In 2005, the production area of sweet pepper in the

  17. Non-destructive quality evaluation of pepper (Capsicum annuum L.) seeds using LED-induced hyperspectral reflectance imaging

    Science.gov (United States)

    In this study, we develop a viability evaluation method for pepper (Capsicum annuum L.) seed based on hyperspectral reflectance imaging. The reflectance spectra of pepper seeds in the 400–700 nm range are collected from hyperspectral reflectance images obtained using blue, green, and red LED illumin...

  18. Solanum americanum: reservoir for Potato virus Y and Cucumber mosaic virus in sweet pepper crops

    Directory of Open Access Journals (Sweden)

    Monika Fecury Moura

    2014-03-01

    Full Text Available Weeds can act as important reservoirs for viruses. Solanum americanum (Black nightshade is a common weed in Brazil and samples showing mosaic were collected from sweet pepper crops to verify the presence of viruses. One sample showed mixed infection between Cucumber mosaic virus (CMV and Potato virus Y (PVY and one sample showed simple infection by PVY. Both virus species were transmitted by plant extract and caused mosaic in tomato (Solanum lycopersicum cv. Santa Clara, sweet pepper (Capsicum annuum cv. Magda, Nicotiana benthamiana and N. tabaccum TNN, and local lesions on Chenopodium quinoa, C. murale and C. amaranticolor. The coat protein sequences for CMV and PVY found in S. americanum are phylogenetically more related to isolates from tomato. We conclude that S. americanum can act as a reservoir for different viruses during and between sweet pepper crop seasons.

  19. Analysis of the blackening of green pepper (Piper nigrum Linnaeus) berries.

    Science.gov (United States)

    Gu, Fenglin; Tan, Lehe; Wu, Huasong; Fang, Yiming; Wang, Qinghuang

    2013-06-01

    This paper investigates polyphenol oxidase (PPO) activity, reduced weight percentage after sun drying, and the changes in colour and appearance of green pepper (Piper nigrum Linnaeus) berries after blanching and sun drying. The results show that the degree of reduced weight percentage and browning in green pepper berries after blanching for 10 min is greater at 100°C than at 90 and 80°C. Moreover, the samples blanched at 100°C for 10 min had the fastest water loss, but the lowest PPO activity. Thus, the PPO enzymatic oxidation of polyphenols might not be the only reason for the browning of green pepper berries. This result is significantly different from that of Variyar, Pendharkar, Banerjeea, and Bandyopadhyay (1988) and therefore deserves further study. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Effect of irradiation on nutritional quality of chicken feet with pickled peppers in shelf life

    International Nuclear Information System (INIS)

    Li Shurong; Zhou Linyan; Yi Jianyong; Feng Min; Li Li; Yang Ping; Wang Dening; Gu Guiqiang; Zhu Jiating

    2013-01-01

    The effect of irradiation on nutritional quality of soft packaged chicken feet with pickled peppers within shelf life were investigated. Chicken feet were irradiated by E-beam which max dose was 10 kGy, and all the samples were stored at 0 ∼ 10℃ and analyzed one month after irradiation treatment. Significant sterilizing effect was got for soft packaged chicken feet with pickled peppers by irradiation; The contents of protein and fat were changed after irradiation treatment; the contents of V_A, V_E and V_B_3 in irradiated chicken feet were increased; The total contents of amino acids increased when irradiation dose were more than 4 kGy; The contents of total acids decreased after irradiation treatment, but the contents of cholesterol and nitrite increased. Above all, irradiation treatment can be used to improve nutritional quality of chicken feet with pickled peppers in shelf life. (authors)

  1. Transverse emittance measurement of high-current single pulse beams using pepper-pot method

    International Nuclear Information System (INIS)

    Ke Jianlin; Zhou Changgeng; Qiu Rui

    2013-01-01

    A pepper pot-imaging plate system has been developed and used to measure the 4-D transverse emittance of a vacuum arc ion source. Single beam pulses of tens to hundreds milliamperes were extracted from the plasma with 64 kV high voltage. An imaging plate was laid after the pepper pot to visualize the ion beamlets passing though the holes on the pepper pot. An application program was developed to show the phase-space distribution and calculate the ellipse and RMS emittances. The normalized RMS emittances are about 6.41 π·mm·mrad in x-direction and 4.61 π·mm·mrad in y-direction. It is shown that the emittance of the vacuum arc ion source is much larger than that of other types of ion sources, which is mainly attributed to the high current and the convex meniscus of this source. (authors)

  2. Sanity of bell pepper fruit fertigated with wastewater from hog production

    Directory of Open Access Journals (Sweden)

    José Antonio Rodrigues de Souza

    2013-08-01

    Full Text Available In order to evaluate the microbiological quality of bell pepper fruits produced with swine wastewater (SW, after preliminary treatment, pepper seedlings of variety Casca Dura Elquida were grown and fertigated with different levels of this wastewater providing 100 and 200% of the amount of the nitrogen needed for cultivation, with and without supplementation of fertilization. The fruits were analyzed for the presence of thermo-tolerant and total coliforms, and of Salmonella sp. The results showed that fertigation with SW, by trickle irrigation system, resulted in bell pepper fruits uncontaminated by fecal coliform and Salmonella spp., in accordance with the microbiological standards required by RDC n°12 02/01/2001.

  3. Natural occurrence of mycotoxins and toxigenic capacity of Alternaria strains from mouldy peppers

    DEFF Research Database (Denmark)

    da Cruz Cabral, Lucía; Terminiello, Laura; Fernández Pinto, Virginia

    2016-01-01

    Sweet pepper (Capsicum annuum) is an important crop cultivated worldwide, with Argentina being one of the major producers in South America. The fruit is susceptible to several fungal diseases, leading to severe economic losses for producers. In this study, Alternaria was found as the prevalent...... genus in mouldy peppers (50% fruits infected). Morphological identification revealed that all 64 Alternaria isolates belonged to small-spored species, most of them corresponding to A. tenuissima, A. arborescens and A. alternata species-groups. Their secondary metabolite profile was evaluated in vitro......; alternariols were synthesized by most of the isolates (91% for alternariol and 92% for alternariol monomethyl ether). A high number of Alternaria spp. also produced tenuazonic acid (64%), altenuene (84%) and tentoxin (72%). In addition, damaged pepper fruits were analysed for the presence of tenuazonic acid...

  4. Rapid biodiesel synthesis from waste pepper seeds without lipid isolation step.

    Science.gov (United States)

    Lee, Jechan; Kim, Jieun; Ok, Yong Sik; Kwon, Eilhann E

    2017-09-01

    In situ transformation of lipid in waste pepper seeds into biodiesel (i.e., fatty acid methyl esters: FAMEs) via thermally-induced transmethylation on silica was mainly investigated in this study. This study reported that waste pepper seeds contained 26.9wt% of lipid and that 94.1% of the total lipid in waste pepper seeds could be converted into biodiesel without lipid extraction step for only ∼1min reaction time. This study also suggested that the optimal temperature for in situ transmethylation was identified as 390°C. Moreover, comparison of in situ process via the conventional transmethylation catalyzed by H 2 SO 4 showed that the introduced biodiesel conversion in this study had a higher tolerance against impurities, thereby being technically feasible. The in situ biodiesel production from other oil-bearing food wastes can be studied. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Foliar Application of the Fungicide Pyraclostrobin Reduced Bacterial Spot Disease of Pepper

    Directory of Open Access Journals (Sweden)

    Beom Ryong Kang

    2018-03-01

    Full Text Available Pyraclostrobin is a broad-spectrum fungicide that inhibits mitochondrial respiration. However, it may also induce systemic resistance effective against bacterial and viral diseases. In this study, we evaluated whether pyraclostrobin enhanced resistance against the bacterial spot pathogen, Xanthomonas euvesicatora on pepper (Capsicum annuum. Although pyraclostrobin alone did not suppressed the in vitro growth of X. euvesicatoria, disease severity in pepper was significantly lower by 69% after treatments with pyraclostrobin alone. A combination of pyraclostrobin with streptomycin reduced disease by over 90% that of the control plants. The preventive control of the pyraclostrobin against bacterial spot was required application 1-3 days before pathogen inoculation. Our findings suggest that the fungicide pyraclostrobin can be used with a chemical pesticide to control bacterial leaf spot diseases in pepper.

  6. Comprehensive study of proteasome inhibitors against Plasmodium falciparum laboratory strains and field isolates from Gabon

    Directory of Open Access Journals (Sweden)

    Kremsner Peter G

    2008-09-01

    Full Text Available Abstract Background The emergence and spread of Plasmodium falciparum resistance to almost all available antimalarial drugs necessitates the search for new chemotherapeutic compounds. The ubiquitin/proteasome system plays a major role in overall protein turnover, especially in fast dividing eukaryotic cells including plasmodia. Previous studies show that the 20S proteasome is expressed and catalytically active in plasmodia and treatment with proteasome inhibitors arrests parasite growth. This is the first comprehensive screening of proteasome inhibitors with different chemical modes of action against laboratory strains of P. falciparum. Subsequently, a selection of inhibitors was tested in field isolates from Lambaréné, Gabon. Methods Epoxomicin, YU101, YU102, MG132, MG115, Z-L3-VS, Ada-Ahx3-L3-VS, lactacystin, bortezomib (Velcade®, gliotoxin, PR11 and PR39 were tested and compared to chloroquine- and artesunate-activities in a standardized in vitro drug susceptibility assay against P. falciparum laboratory strains 3D7, D10 and Dd2. Freshly obtained field isolates from Lambaréné, Gabon, were used to measure the activity of chloroquine, artesunate, epoxomicin, MG132, lactacystin and bortezomib. Parasite growth was detected through histidine-rich protein 2 (HRP2 production. Raw data were fitted by a four-parameter logistic model and individual inhibitory concentrations (50%, 90%, and 99% were calculated. Results Amongst all proteasome inhibitors tested, epoxomicin showed the highest activity in chloroquine-susceptible (IC50: 6.8 nM [3D7], 1.7 nM [D10] and in chloroquine-resistant laboratory strains (IC50: 10.4 nM [Dd2] as well as in field isolates (IC50: 8.5 nM. The comparator drug artesunate was even more active (IC50: 1.0 nM, whereas all strains were chloroquine-resistant (IC50: 113 nM. Conclusion The peptide α',β'-epoxyketone epoxomicin is highly active against P. falciparum regardless the grade of the parasite's chloroquine

  7. Physiology of pepper fruit and the metabolism of antioxidants: chloroplasts, mitochondria and peroxisomes

    Science.gov (United States)

    Palma, José M.; Sevilla, Francisca; Jiménez, Ana; del Río, Luis A.; Corpas, Francisco J.; Álvarez de Morales, Paz; Camejo, Daymi M.

    2015-01-01

    Background and Aims Pepper (Capsicum annuum) contains high levels of antioxidants, such as vitamins A and C and flavonoids. However, information on the role of these beneficial compounds in the physiology of pepper fruit remains scarce. Recent studies have shown that antioxidants in ripe pepper fruit play a key role in responses to temperature changes, and the redox state at the time of harvest affects the nutritional value for human consumption. In this paper, the role of antioxidant metabolism of pepper fruit during ripening and in the response to low temperature is addressed, paying particular attention to ascorbate, NADPH and the superoxide dismutase enzymatic system. The participation of chloroplasts, mitochondria and peroxisomes in the ripening process is also investigated. Scope and Results Important changes occur at a subcellular level during ripening of pepper fruit. Chloroplasts turn into chromoplasts, with drastic conversion of their metabolism, and the role of the ascorbate–glutathione cycle is essential. In mitochondria from red fruits, higher ascorbate peroxidase (APX) and Mn-SOD activities are involved in avoiding the accumulation of reactive oxygen species in these organelles during ripening. Peroxisomes, whose antioxidant capacity at fruit ripening is substantially affected, display an atypical metabolic pattern during this physiological stage. In spite of these differences observed in the antioxidative metabolism of mitochondria and peroxisomes, proteomic analysis of these organelles, carried out by 2-D electrophoresis and MALDI-TOF/TOF and provided here for the first time, reveals no changes between the antioxidant metabolism from immature (green) and ripe (red) fruits. Conclusions Taken together, the results show that investigation of molecular and enzymatic antioxidants from cell compartments, especially chloroplasts, mitochondria and peroxisomes, is a useful tool to study the physiology of pepper fruit, particularly in the context of

  8. Constitutive expression of a fungus-inducible carboxylesterase improves disease resistance in transgenic pepper plants.

    Science.gov (United States)

    Ko, Moonkyung; Cho, Jung Hyun; Seo, Hyo-Hyoun; Lee, Hyun-Hwa; Kang, Ha-Young; Nguyen, Thai Son; Soh, Hyun Cheol; Kim, Young Soon; Kim, Jeong-Il

    2016-08-01

    Resistance against anthracnose fungi was enhanced in transgenic pepper plants that accumulated high levels of a carboxylesterase, PepEST in anthracnose-susceptible fruits, with a concurrent induction of antioxidant enzymes and SA-dependent PR proteins. A pepper esterase gene (PepEST) is highly expressed during the incompatible interaction between ripe fruits of pepper (Capsicum annuum L.) and a hemibiotrophic anthracnose fungus (Colletotrichum gloeosporioides). In this study, we found that exogenous application of recombinant PepEST protein on the surface of the unripe pepper fruits led to a potentiated state for disease resistance in the fruits, including generation of hydrogen peroxide and expression of pathogenesis-related (PR) genes that encode mostly small proteins with antimicrobial activity. To elucidate the role of PepEST in plant defense, we further developed transgenic pepper plants overexpressing PepEST under the control of CaMV 35S promoter. Molecular analysis confirmed the establishment of three independent transgenic lines carrying single copy of transgenes. The level of PepEST protein was estimated to be approximately 0.002 % of total soluble protein in transgenic fruits. In response to the anthracnose fungus, the transgenic fruits displayed higher expression of PR genes, PR3, PR5, PR10, and PepThi, than non-transgenic control fruits did. Moreover, immunolocalization results showed concurrent localization of ascorbate peroxidase (APX) and PR3 proteins, along with the PepEST protein, in the infected region of transgenic fruits. Disease rate analysis revealed significantly low occurrence of anthracnose disease in the transgenic fruits, approximately 30 % of that in non-transgenic fruits. Furthermore, the transgenic plants also exhibited resistance against C. acutatum and C. coccodes. Collectively, our results suggest that overexpression of PepEST in pepper confers enhanced resistance against the anthracnose fungi by activating the defense signaling

  9. Gambogic Acid Is a Tissue-Specific Proteasome Inhibitor In Vitro and In Vivo

    Directory of Open Access Journals (Sweden)

    Xiaofen Li

    2013-01-01

    Full Text Available Gambogic acid (GA is a natural compound derived from Chinese herbs that has been approved by the Chinese Food and Drug Administration for clinical trials in cancer patients; however, its molecular targets have not been thoroughly studied. Here, we report that GA inhibits tumor proteasome activity, with potency comparable to bortezomib but much less toxicity. First, GA acts as a prodrug and only gains proteasome-inhibitory function after being metabolized by intracellular CYP2E1. Second, GA-induced proteasome inhibition is a prerequisite for its cytotoxicity and anticancer effect without off-targets. Finally, because expression of the CYP2E1 gene is very high in tumor tissues but low in many normal tissues, GA could therefore produce tissue-specific proteasome inhibition and tumor-specific toxicity, with clinical significance for designing novel strategies for cancer treatment.

  10. Oxidized SOD1 alters proteasome activities in vitro and in the cortex of SOD1 overexpressing mice.

    Science.gov (United States)

    Le Pecheur, Marie; Bourdon, Emmanuel; Paly, Evelyne; Farout, Luc; Friguet, Bertrand; London, Jacqueline

    2005-07-04

    Premature ageing, one of the characteristics of Down syndrome (DS), may involve oxidative stress and impairment of proteasome activity. Transgenic mice overexpressing the human copper/zinc superoxide dismutase (SOD1) gene are one of the first murine models for DS and it has been shown that SOD1 overexpression might be either deleterious or beneficial. Here, we show a reduction in proteasome activities in the cortex of SOD1 transgenic mice and an associated increase in the content of oxidized SOD1 protein. As we demonstrate that in vitro oxidized SOD can inhibit purified proteasome peptidase activities, modified SOD1 might be partially responsible for proteasome inhibition shown in SOD1 transgenic mice.

  11. Hybrid breeding in pepper (capsicum annuum L. ). [Gamma and x radiation

    Energy Technology Data Exchange (ETDEWEB)

    Milkova, L; Daskalov, S [Akademiya na Selskostopanskite Nauki, Sofia (Bulgaria). Inst. po Genetika

    1981-01-01

    Male sterile forms of peppers with gene sterility were produced by ..gamma..- and X-irradiation of dry seeds of cv. Pazarjishka kapiya 794 and Zlaten medal and subsequent selection in M/sub 2/. Male sterile lines suitable for developing hybrid cultivars intended for various kinds of production were obtained by backcrossing with lines and cultivars having valuable economic characters and high combining ability. The promising pepper Belasitsa, Prista, Lyulin and Strouma (all of them for early field production), and Izoumroud and Prevuzhoden (for glasshouse production) are described.

  12. Simple emittance measurement of negative hydrogen ion beam using pepper-pot method

    International Nuclear Information System (INIS)

    Hamabe, M.; Tsumori, K.; Takeiri, Y.; Kaneko, O.; Asano, E.; Kawamoto, T.; Kuroda, T.; Guharay, S.K.

    1997-01-01

    A simple apparatus for emittance measurement using pepper-pot method is developed. The pepper-pot patterns are directly exposed and recorded on a Kapton foil. Using this apparatus, emittance was measured in the case of the negative hydrogen (H - ) beam from the large negative ion source, which is the 1/3 scaled test device for the negative-ion-based neutral beam injection (N-NBI) on the Large Helical Device (LHD). As the consequence of the first trial, the 95% normalized emittance value is measured as 0.59 mm mrad. (author)

  13. Influence of Copigment derived from Tasmannia Pepper Leaves on Davidson’s Plum Anthocyanins

    DEFF Research Database (Denmark)

    Jensen, Morten Busch; López-de-Dicastillo Bergamo, Carolina Ana; Payet, René Marc

    2011-01-01

    . An addition of a variety of phenolic acids significantly increased color intensity indicating the formation of copigmentation complexes. Commercial chlorogenic acid as well as extract from a native Australian herb rich in chlorogenic acid, Tasmannia pepper leaf (Tasmannia lanceolata, R. Br.), were both tested...... in model soft drink solutions subjected to light irradiation and heat treatment. In both cases the addition of the copigment resulted in a lasting increase in color intensity. In conclusion, Davidson’s plum extract can successfully be utilized as a source of natural food color. Extract from Tasmania pepper...

  14. Rheological behaviour of white pepper gels - a new method for studying the effect of irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Esteves, M. Paula E-mail: paula.esteves@ineti.pt; Raymundo, Anabela; Sousa, Isabel de; Andrade, M. Eduarda; Empis, Jose

    2002-07-01

    Ground white pepper was treated by gamma radiation at average doses of 5, 10 and 15 kGy and the alterations in the rheological properties of its aqueous suspensions were used to study the radiation effects. Amylose and amylopectin suspensions were used as model systems. Ground pepper gels presented decreased strength expressed by a decrease of G' and G'' values and smaller limiting viscosity ({eta}{sub 0}), as the irradiation dose increased. A similar behaviour was shown by the amylopectin gels. For the amylose gels, in opposition, irradiation induced a higher level of gel structure.

  15. Consumers’ Preference for Sweet Peppers with different Process Attributes: A Discrete Choice Experiment in Taiwan

    Directory of Open Access Journals (Sweden)

    Ching-Hua Yeh

    2016-08-01

    Full Text Available Based on an online discrete choice experiment (DCE this study investigates the relative importance of food label information (country of origin, production methods, chemical residue testing (CRT and price for Taiwanese consumers’ in their purchase of sweet peppers. Results show that respondents focus mostly on the COO labeling during their sweet-pepper shopping, followed by price. Information concerning CRT results and production methods are of less importance. Our findings also indicate that interaction between attributes matter and that preference for attribute levels differs depending on socioeconomic characteristics.

  16. Preliminary examination of induced radio activity in pepper by 10 MeV electron irradiation

    International Nuclear Information System (INIS)

    Furuta, Masakazu; Katayama, Tadashi; Ito, Norio; Mizohata, Akira; Matsunami, Tadao; Toratani, Hirokazu; Takeda, Atsuhiko

    1989-01-01

    β-ray measurement was performed on 10 MeV electron-irradiated black pepper and white pepper in order to reconfirm the wholesomeness of irradiated food and present unambiguous data to general consumers concerning about the induced radioactivity in the irradiated foods. From elemental composition of the samples and investigation of photonuclear reactions, several β-emmitters were listed up. But no radioactivity other than from natural sources was detected in the irradiated sample by β-ray counting with 2 π gass flow counter, suggesting that the induced β-emmitters in the irradiated sample was below the detection limit of its induced radioactivity. (author)

  17. Rheological behaviour of white pepper gels - a new method for studying the effect of irradiation

    International Nuclear Information System (INIS)

    Esteves, M. Paula; Raymundo, Anabela; Sousa, Isabel de; Andrade, M. Eduarda; Empis, Jose

    2002-01-01

    Ground white pepper was treated by gamma radiation at average doses of 5, 10 and 15 kGy and the alterations in the rheological properties of its aqueous suspensions were used to study the radiation effects. Amylose and amylopectin suspensions were used as model systems. Ground pepper gels presented decreased strength expressed by a decrease of G' and G'' values and smaller limiting viscosity (η 0 ), as the irradiation dose increased. A similar behaviour was shown by the amylopectin gels. For the amylose gels, in opposition, irradiation induced a higher level of gel structure

  18. Simple emittance measurement of negative hydrogen ion beam using pepper-pot method

    Energy Technology Data Exchange (ETDEWEB)

    Hamabe, M.; Tsumori, K.; Takeiri, Y.; Kaneko, O.; Asano, E.; Kawamoto, T.; Kuroda, T. [National Inst. for Fusion Science, Nagoya (Japan); Guharay, S.K.

    1997-02-01

    A simple apparatus for emittance measurement using pepper-pot method is developed. The pepper-pot patterns are directly exposed and recorded on a Kapton foil. Using this apparatus, emittance was measured in the case of the negative hydrogen (H{sup -}) beam from the large negative ion source, which is the 1/3 scaled test device for the negative-ion-based neutral beam injection (N-NBI) on the Large Helical Device (LHD). As the consequence of the first trial, the 95% normalized emittance value is measured as 0.59 mm mrad. (author)

  19. Resolution of "salt and pepper" appearance of the skull with vitamin D therapy

    Directory of Open Access Journals (Sweden)

    Gursimran Kaur

    2013-01-01

    Full Text Available Chronic hypovitaminosis D leads to state of decreased mineralization and generalized osteomalacia. It also results in secondary hyperparathyroidism causing increased bone turn over and decreased bone mass, manifested radiologically as a "salt and pepper" appearance in skull, subperiosteal resorption, bone cysts and lytic lesions. In this case, a young male patient with hypovitaminosis D and secondary hyperparathyroidism, radiological features show resolution of "salt and pepper" appearance of the skull with vitamin D in 11 months and regression of other lytic lesions.

  20. Phylogenetic relationships, diversification and expansion of chili peppers (Capsicum, Solanaceae)

    Science.gov (United States)

    Carrizo García, Carolina; Barfuss, Michael H. J.; Sehr, Eva M.; Barboza, Gloria E.; Samuel, Rosabelle; Moscone, Eduardo A.; Ehrendorfer, Friedrich

    2016-01-01

    Background and Aims Capsicum (Solanaceae), native to the tropical and temperate Americas, comprises the well-known sweet and hot chili peppers and several wild species. So far, only partial taxonomic and phylogenetic analyses have been done for the genus. Here, the phylogenetic relationships between nearly all taxa of Capsicum were explored to test the monophyly of the genus and to obtain a better knowledge of species relationships, diversification and expansion. Methods Thirty-four of approximately 35 Capsicum species were sampled. Maximum parsimony and Bayesian inference analyses were performed using two plastid markers (matK and psbA-trnH) and one single-copy nuclear gene (waxy). The evolutionary changes of nine key features were reconstructed following the parsimony ancestral states method. Ancestral areas were reconstructed through a Bayesian Markov chain Monte Carlo analysis. Key Results Capsicum forms a monophyletic clade, with Lycianthes as a sister group, following both phylogenetic approaches. Eleven well-supported clades (four of them monotypic) can be recognized within Capsicum, although some interspecific relationships need further analysis. A few features are useful to characterize different clades (e.g. fruit anatomy, chromosome base number), whereas some others are highly homoplastic (e.g. seed colour). The origin of Capsicum is postulated in an area along the Andes of western to north-western South America. The expansion of the genus has followed a clockwise direction around the Amazon basin, towards central and south-eastern Brazil, then back to western South America, and finally northwards to Central America. Conclusions New insights are provided regarding interspecific relationships, character evolution, and geographical origin and expansion of Capsicum. A clearly distinct early-diverging clade can be distinguished, centred in western–north-western South America. Subsequent rapid speciation has led to the origin of the remaining clades. The

  1. The 11S Proteasomal Activator REGγ Impacts Polyglutamine-Expanded Androgen Receptor Aggregation and Motor Neuron Viability through Distinct Mechanisms

    Directory of Open Access Journals (Sweden)

    Jill M. Yersak

    2017-05-01

    Full Text Available Spinal and bulbar muscular atrophy (SBMA is caused by expression of a polyglutamine (polyQ-expanded androgen receptor (AR. The inefficient nuclear proteasomal degradation of the mutant AR results in the formation of nuclear inclusions containing amino-terminal fragments of the mutant AR. PA28γ (also referred to as REGγ is a nuclear 11S-proteasomal activator with limited proteasome activation capabilities compared to its cytoplasmic 11S (PA28α, PA28β counterparts. To clarify the role of REGγ in polyQ-expanded AR metabolism, we carried out genetic and biochemical studies in cell models of SBMA. Overexpression of REGγ in a PC12 cell model of SBMA increased polyQ-expanded AR aggregation and contributed to polyQ-expanded AR toxicity in the presence of dihydrotestosterone (DHT. These effects of REGγ were independent of its association with the proteasome and may be due, in part, to the decreased binding of polyQ-expanded AR by the E3 ubiquitin-ligase MDM2. Unlike its effects in PC12 cells, REGγ overexpression rescued transgenic SBMA motor neurons from DHT-induced toxicity in a proteasome binding-dependent manner, suggesting that the degradation of a specific 11S proteasome substrate or substrates promotes motor neuron viability. One potential substrate that we found to play a role in mutant AR toxicity is the splicing factor SC35. These studies reveal that, depending on the cellular context, two biological roles for REGγ impact cell viability in the face of polyQ-expanded AR; a proteasome binding-independent mechanism directly promotes mutant AR aggregation while a proteasome binding-dependent mechanism promotes cell viability. The balance between these functions likely determines REGγ effects on polyQ-expanded AR-expressing cells.

  2. Proteasome activity is important for replication recovery, CHK1 phosphorylation and prevention of G2 arrest after low-dose formaldehyde

    Energy Technology Data Exchange (ETDEWEB)

    Ortega-Atienza, Sara; Green, Samantha E.; Zhitkovich, Anatoly, E-mail: anatoly_zhitkovich@brown.edu

    2015-07-15

    Formaldehyde (FA) is a human carcinogen with numerous sources of environmental and occupational exposures. This reactive aldehyde is also produced endogenously during metabolism of drugs and other processes. DNA–protein crosslinks (DPCs) are considered to be the main genotoxic lesions for FA. Accumulating evidence suggests that DPC repair in high eukaryotes involves proteolysis of crosslinked proteins. Here, we examined a role of the main cellular proteolytic machinery proteasomes in toxic responses of human lung cells to low FA doses. We found that transient inhibition of proteasome activity increased cytotoxicity and diminished clonogenic viability of FA-treated cells. Proteasome inactivation exacerbated suppressive effects of FA on DNA replication and increased the levels of the genotoxic stress marker γ-H2AX in normal human cells. A transient loss of proteasome activity in FA-exposed cells also caused delayed perturbations of cell cycle, which included G2 arrest and a depletion of S-phase populations at FA doses that had no effects in control cells. Proteasome activity diminished p53-Ser15 phosphorylation but was important for FA-induced CHK1 phosphorylation, which is a biochemical marker of DPC proteolysis in replicating cells. Unlike FA, proteasome inhibition had no effect on cell survival and CHK1 phosphorylation by the non-DPC replication stressor hydroxyurea. Overall, we obtained evidence for the importance of proteasomes in protection of human cells against biologically relevant doses of FA. Biochemically, our findings indicate the involvement of proteasomes in proteolytic repair of DPC, which removes replication blockage by these highly bulky lesions. - Highlights: • Proteasome inhibition enhances cytotoxicity of low-dose FA in human lung cells. • Active proteasomes diminish replication-inhibiting effects of FA. • Proteasome activity prevents delayed G2 arrest in FA-treated cells. • Proteasome inhibition exacerbates replication stress by FA in

  3. BRK targets Dok1 for ubiquitin-mediated proteasomal degradation to promote cell proliferation and migration.

    Directory of Open Access Journals (Sweden)

    Sayem Miah

    Full Text Available Breast tumor kinase (BRK, also known as protein tyrosine kinase 6 (PTK6, is a non-receptor tyrosine kinase overexpressed in more that 60% of human breast carcinomas. The overexpression of BRK has been shown to sensitize mammary epithelial cells to mitogenic signaling and to promote cell proliferation and tumor formation. The molecular mechanisms of BRK have been unveiled by the identification and characterization of BRK target proteins. Downstream of tyrosine kinases 1 or Dok1 is a scaffolding protein and a substrate of several tyrosine kinases. Herein we show that BRK interacts with and phosphorylates Dok1 specifically on Y362. We demonstrate that this phosphorylation by BRK significantly downregulates Dok1 in a ubiquitin-proteasome-mediated mechanism. Together, these results suggest a novel mechanism of action of BRK in the promotion of tumor formation, which involves the targeting of tumor suppressor Dok1 for degradation through the ubiquitin proteasomal pathway.

  4. BRK targets Dok1 for ubiquitin-mediated proteasomal degradation to promote cell proliferation and migration.

    Science.gov (United States)

    Miah, Sayem; Goel, Raghuveera Kumar; Dai, Chenlu; Kalra, Natasha; Beaton-Brown, Erika; Bagu, Edward T; Bonham, Keith; Lukong, Kiven E

    2014-01-01

    Breast tumor kinase (BRK), also known as protein tyrosine kinase 6 (PTK6), is a non-receptor tyrosine kinase overexpressed in more that 60% of human breast carcinomas. The overexpression of BRK has been shown to sensitize mammary epithelial cells to mitogenic signaling and to promote cell proliferation and tumor formation. The molecular mechanisms of BRK have been unveiled by the identification and characterization of BRK target proteins. Downstream of tyrosine kinases 1 or Dok1 is a scaffolding protein and a substrate of several tyrosine kinases. Herein we show that BRK interacts with and phosphorylates Dok1 specifically on Y362. We demonstrate that this phosphorylation by BRK significantly downregulates Dok1 in a ubiquitin-proteasome-mediated mechanism. Together, these results suggest a novel mechanism of action of BRK in the promotion of tumor formation, which involves the targeting of tumor suppressor Dok1 for degradation through the ubiquitin proteasomal pathway.

  5. Retinoblastoma protein co-purifies with proteasomal insulin-degrading enzyme: Implications for cell proliferation control

    Energy Technology Data Exchange (ETDEWEB)

    Radulescu, Razvan T., E-mail: ratura@gmx.net [Molecular Concepts Research (MCR), Muenster (Germany); Duckworth, William C. [Department of Medicine, Phoenix VA Health Care System, Phoenix, AZ (United States); Levy, Jennifer L. [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States); Fawcett, Janet, E-mail: janet.fawcett@va.gov [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States)

    2010-04-30

    Previous investigations on proteasomal preparations containing insulin-degrading enzyme (IDE; EC 3.4.24.56) have invariably yielded a co-purifying protein with a molecular weight of about 110 kDa. We have now found both in MCF-7 breast cancer and HepG2 hepatoma cells that this associated molecule is the retinoblastoma tumor suppressor protein (RB). Interestingly, the amount of RB in this protein complex seemed to be lower in HepG2 vs. MCF-7 cells, indicating a higher (cytoplasmic) protein turnover in the former vs. the latter cells. Moreover, immunofluorescence showed increased nuclear localization of RB in HepG2 vs. MCF-7 cells. Beyond these subtle differences between these distinct tumor cell types, our present study more generally suggests an interplay between RB and IDE within the proteasome that may have important growth-regulatory consequences.

  6. Gammaherpesviral Tegument Proteins, PML-Nuclear Bodies and the Ubiquitin-Proteasome System

    Directory of Open Access Journals (Sweden)

    Florian Full

    2017-10-01

    Full Text Available Gammaherpesviruses like Epstein-Barr virus (EBV and Kaposi’s sarcoma-associated herpesvirus (KSHV subvert the ubiquitin proteasome system for their own benefit in order to facilitate viral gene expression and replication. In particular, viral tegument proteins that share sequence homology to the formylglycineamide ribonucleotide amidotransferase (FGARAT, or PFAS, an enzyme in the cellular purine biosynthesis, are important for disrupting the intrinsic antiviral response associated with Promyelocytic Leukemia (PML protein-associated nuclear bodies (PML-NBs by proteasome-dependent and independent mechanisms. In addition, all herpesviruses encode for a potent ubiquitin protease that can efficiently remove ubiquitin chains from proteins and thereby interfere with several different cellular pathways. In this review, we discuss mechanisms and functional consequences of virus-induced ubiquitination and deubiquitination for early events in gammaherpesviral infection.

  7. Mitochondrial and Ubiquitin Proteasome System Dysfunction in Ageing and Disease: Two Sides of the Same Coin?

    Directory of Open Access Journals (Sweden)

    Jaime M. Ross

    2015-08-01

    Full Text Available Mitochondrial dysfunction and impairment of the ubiquitin proteasome system have been described as two hallmarks of the ageing process. Additionally, both systems have been implicated in the etiopathogenesis of many age-related diseases, particularly neurodegenerative disorders, such as Alzheimer’s and Parkinson’s disease. Interestingly, these two systems are closely interconnected, with the ubiquitin proteasome system maintaining mitochondrial homeostasis by regulating organelle dynamics, the proteome, and mitophagy, and mitochondrial dysfunction impairing cellular protein homeostasis by oxidative damage. Here, we review the current literature and argue that the interplay of the two systems should be considered in order to better understand the cellular dysfunction observed in ageing and age-related diseases. Such an approach may provide valuable insights into molecular mechanisms underlying the ageing process, and further discovery of treatments to counteract ageing and its associated diseases. Furthermore, we provide a hypothetical model for the heterogeneity described among individuals during ageing.

  8. SOCS-1 Localizes to the Microtubule Organizing Complex-Associated 20S Proteasome

    OpenAIRE

    Vuong, Bao Q.; Arenzana, Teresita L.; Showalter, Brian M.; Losman, Julie; Chen, X. Peter; Mostecki, Justin; Banks, Alexander S.; Limnander, Andre; Fernandes, Neil; Rothman, Paul B.

    2005-01-01

    The regulation of cytokine signaling is critical for controlling cellular proliferation and activation during an immune response. SOCS-1 is a potent inhibitor of Jak kinase activity and of signaling initiated by several cytokines. SOCS-1 protein levels are tightly regulated, and recent data suggest that SOCS-1 may regulate the protein levels of some signaling proteins by the ubiquitin proteasome pathway; however, the cellular mechanism by which SOCS-1 directs proteins for degradation is unkno...

  9. Proteasome-independent degradation of HIV-1 in naturally non-permissive human placental trophoblast cells

    Directory of Open Access Journals (Sweden)

    Barré-Sinoussi Françoise

    2009-05-01

    Full Text Available Abstract Background The human placenta-derived cell line BeWo has been demonstrated to be restrictive to cell-free HIV-1 infection. BeWo cells are however permissive to infection by VSV-G pseudotyped HIV-1, which enters cells by a receptor-independent mechanism, and to infection by HIV-1 via a cell-to-cell route. Results Here we analysed viral entry in wild type BeWo (CCR5+, CXCR4+ and BeWo-CD4+ (CD4+, CCR5+, CXCR4+ cells. We report that HIV-1 internalisation is not restricted in either cell line. Levels of internalised p24 antigen between VSV-G HIV-1 pseudotypes and R5 or X4 virions were comparable. We next analysed the fate of internalised virions; X4 and R5 HIV-1 virions were less stable over time in BeWo cells than VSV-G HIV-1 pseudotypes. We then investigated the role of the proteasome in restricting cell-free HIV-1 infection in BeWo cells using proteasome inhibitors. We observed an increase in the levels of VSV-G pseudotyped HIV-1 infection in proteasome-inhibitor treated cells, but the infection by R5-Env or X4-Env pseudotyped virions remains restricted. Conclusion Collectively these results suggest that cell-free HIV-1 infection encounters a surface block leading to a non-productive entry route, which either actively targets incoming virions for non-proteasomal degradation, and impedes their release into the cytoplasm, or causes the inactivation of mechanisms essential for viral replication.

  10. CCR 20th anniversary commentary: Preclinical study of proteasome inhibitor bortezomib in head and neck cancer.

    Science.gov (United States)

    Allen, Clint T; Conley, Barbara; Sunwoo, John B; Van Waes, Carter

    2015-03-01

    In a study published in the May 1, 2001, issue of Clinical Cancer Research, Sunwoo and colleagues provided evidence for proteasome inhibition of NF-κB and tumorigenesis, supporting early-phase clinical trials in solid malignancies of the upper aerodigestive tract. Subsequent clinical studies uncovered a dichotomy of responses in patients with hematopoietic and solid malignancies, and the mechanisms of resistance. ©2015 American Association for Cancer Research.

  11. Proteasomal degradation of ubiquitinated proteins in oocyte meiosis and fertilization in mammals

    Czech Academy of Sciences Publication Activity Database

    Karabínová, Pavla; Kubelka, Michal; Šušor, Andrej

    2011-01-01

    Roč. 346, č. 1 (2011), s. 1-9 ISSN 0302-766X R&D Projects: GA ČR GAP502/10/0944; GA ČR(CZ) GD204/09/H084 Institutional research plan: CEZ:AV0Z50450515 Keywords : Oocyte * Proteasome * Meiosis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.114, year: 2011

  12. Bcl2-associated Athanogene 3 Interactome Analysis Reveals a New Role in Modulating Proteasome Activity*

    Science.gov (United States)

    Chen, Ying; Yang, Li-Na; Cheng, Li; Tu, Shun; Guo, Shu-Juan; Le, Huang-Ying; Xiong, Qian; Mo, Ran; Li, Chong-Yang; Jeong, Jun-Seop; Jiang, Lizhi; Blackshaw, Seth; Bi, Li-Jun; Zhu, Heng; Tao, Sheng-Ce; Ge, Feng

    2013-01-01

    Bcl2-associated athanogene 3 (BAG3), a member of the BAG family of co-chaperones, plays a critical role in regulating apoptosis, development, cell motility, autophagy, and tumor metastasis and in mediating cell adaptive responses to stressful stimuli. BAG3 carries a BAG domain, a WW domain, and a proline-rich repeat (PXXP), all of which mediate binding to different partners. To elucidate BAG3's interaction network at the molecular level, we employed quantitative immunoprecipitation combined with knockdown and human proteome microarrays to comprehensively profile the BAG3 interactome in humans. We identified a total of 382 BAG3-interacting proteins with diverse functions, including transferase activity, nucleic acid binding, transcription factors, proteases, and chaperones, suggesting that BAG3 is a critical regulator of diverse cellular functions. In addition, we characterized interactions between BAG3 and some of its newly identified partners in greater detail. In particular, bioinformatic analysis revealed that the BAG3 interactome is strongly enriched in proteins functioning within the proteasome-ubiquitination process and that compose the proteasome complex itself, suggesting that a critical biological function of BAG3 is associated with the proteasome. Functional studies demonstrated that BAG3 indeed interacts with the proteasome and modulates its activity, sustaining cell survival and underlying resistance to therapy through the down-modulation of apoptosis. Taken as a whole, this study expands our knowledge of the BAG3 interactome, provides a valuable resource for understanding how BAG3 affects different cellular functions, and demonstrates that biologically relevant data can be harvested using this kind of integrated approach. PMID:23824909

  13. Analyzing proteasomal subunit expression reveals Rpt4 as a prognostic marker in stage II colorectal cancer.

    LENUS (Irish Health Repository)

    2012-02-01

    Colorectal cancer is a leading cause of cancer-related deaths worldwide. Early diagnosis and treatment of colorectal cancer is the key to improving survival rates and as such a need exists to identify patients who may benefit from adjuvant chemotherapy. The dysregulation of the ubiquitin-proteasome system (UPS) has been implicated in oncogenesis and cancer cell survival, and proteasome inhibitors are in clinical use for a number of malignancies including multiple myeloma. In our study, we examined the protein expression of several key components of the UPS in colorectal cancer using immunohistochemistry to determine expression levels of ubiquitinylated proteins and the proteasomal subunits, 20S core and Rpt4 in a cohort of 228 patients with colon cancer. Multivariate Cox analysis revealed that neither the intensity of either ubiquitinylated proteins or the 20S core was predictive in either Stage II or III colon cancer for disease free survival or overall survival. In contrast, in Stage II patients increased Rpt4 staining was significantly associated with disease free survival (Cox proportional hazard ratio 0.605; p = 0.0217). Our data suggest that Rpt4 is an independent prognostic variable for Stage II colorectal cancer and may aid in the decision of which patients undergo adjuvant chemotherapy.

  14. VRK1 regulates Cajal body dynamics and protects coilin from proteasomal degradation in cell cycle.

    Science.gov (United States)

    Cantarero, Lara; Sanz-García, Marta; Vinograd-Byk, Hadar; Renbaum, Paul; Levy-Lahad, Ephrat; Lazo, Pedro A

    2015-06-12

    Cajal bodies (CBs) are nuclear organelles associated with ribonucleoprotein functions and RNA maturation. CBs are assembled on coilin, its main scaffold protein, in a cell cycle dependent manner. The Ser-Thr VRK1 (vaccinia-related kinase 1) kinase, whose activity is also cell cycle regulated, interacts with and phosphorylates coilin regulating assembly of CBs. Coilin phosphorylation is not necessary for its interaction with VRK1, but it occurs in mitosis and regulates coilin stability. Knockdown of VRK1 or VRK1 inactivation by serum deprivation causes a loss of coilin phosphorylation in Ser184 and of CBs formation, which are rescued with an active VRK1, but not by kinase-dead VRK1. The phosphorylation of coilin in Ser184 occurs during mitosis before assembly of CBs. Loss of coilin phosphorylation results in disintegration of CBs, and of coilin degradation that is prevented by proteasome inhibitors. After depletion of VRK1, coilin is ubiquitinated in nuclei, which is partly mediated by mdm2, but its proteasomal degradation occurs in cytosol and is prevented by blocking its nuclear export. We conclude that VRK1 is a novel regulator of CBs dynamics and stability in cell cycle by protecting coilin from ubiquitination and degradation in the proteasome, and propose a model of CB dynamics.

  15. Alterations of the intracellular peptidome in response to the proteasome inhibitor bortezomib.

    Directory of Open Access Journals (Sweden)

    Julia S Gelman

    Full Text Available Bortezomib is an antitumor drug that competitively inhibits proteasome beta-1 and beta-5 subunits. While the impact of bortezomib on protein stability is known, the effect of this drug on intracellular peptides has not been previously explored. A quantitative peptidomics technique was used to examine the effect of treating human embryonic kidney 293T (HEK293T cells with 5-500 nM bortezomib for various lengths of time (30 minutes to 16 hours, and human neuroblastoma SH-SY5Y cells with 500 nM bortezomib for 1 hour. Although bortezomib treatment decreased the levels of some intracellular peptides, the majority of peptides were increased by 50-500 nM bortezomib. Peptides requiring cleavage at acidic and hydrophobic sites, which involve beta-1 and -5 proteasome subunits, were among those elevated by bortezomib. In contrast, the proteasome inhibitor epoxomicin caused a decrease in the levels of many of these peptides. Although bortezomib can induce autophagy under certain conditions, the rapid bortezomib-mediated increase in peptide levels did not correlate with the induction of autophagy. Taken together, the present data indicate that bortezomib alters the balance of intracellular peptides, which may contribute to the biological effects of this drug.

  16. What do we really know about the ubiquitin-proteasome pathway in muscle atrophy?

    Science.gov (United States)

    Jagoe, R. T.; Goldberg, A. L.

    2001-01-01

    Studies of many different rodent models of muscle wasting have indicated that accelerated proteolysis via the ubiquitin-proteasome pathway is the principal cause of muscle atrophy induced by fasting, cancer cachexia, metabolic acidosis, denervation, disuse, diabetes, sepsis, burns, hyperthyroidism and excess glucocorticoids. However, our understanding about how muscle proteins are degraded, and how the ubiquitin-proteasome pathway is activated in muscle under these conditions, is still very limited. The identities of the important ubiquitin-protein ligases in skeletal muscle, and the ways in which they recognize substrates are still largely unknown. Recent in-vitro studies have suggested that one set of ubquitination enzymes, E2(14K) and E3(alpha), which are responsible for the 'N-end rule' system of ubiquitination, plays an important role in muscle, especially in catabolic states. However, their functional significance in degrading different muscle proteins is still unclear. This review focuses on the many gaps in our understanding of the functioning of the ubiquitin-proteasome pathway in muscle atrophy, and highlights the strengths and limitations of the different experimental approaches used in such studies.

  17. Isolation, characterization and identification of pericarp-degrading bacteria for the production of off-odour-free white pepper from fresh berries of Piper nigrum L.

    Science.gov (United States)

    Vinod, V; Kumar, A; Zachariah, T J

    2014-04-01

    To isolate, fermentatively evaluate and identify black pepper (Piper nigrum L.)-associated bacteria for the microbial decortication of fresh ripened berries and dried black pepper for preparation of off-odour-free white pepper. Among 45 bacterial isolates obtained from black pepper, seven of them were found to decorticate black pepper (>60%) and fresh pepper berries (98-100%) into white pepper within 5 days of immersion in bacterial suspension. The 16S rRNA genes (1500-bp amplicon) of these bacteria were sequenced, and species identity was established by closest match in GenBank. Superior-quality white pepper was obtained with Bacillus subtilis (IISR WP 33, 34, 38), Bacillus licheniformis (IISR WP 43), Acinetobacter baumanii (IISR WP 35), Klebsiella pneumoniae (IISR WP 19) and Microbacterium barkeri (IISR WP25). The bacterial isolates were found to secrete multiple hydrolytic enzymes such as cellulase, pectinase, amylase, protease and xylanase. Bacterial cultures were deposited with International Depository Authority at Microbial Type Culture Collection, India, as patent deposits as prescribed in Budapest Treaty for microbial deposits. The white pepper, thus obtained from bacterial decortication process, was free from off-odour compound, especially skatole. Other biochemical constituents such as oleoresin, piperine and essential oils were found in the acceptable range. The bacterial decortication did not affect inherent constituents of pepper such as essential oil constituents, oleoresin and piperine content. One of the most significant findings of the work is identification of specific bacterial species for decortication of fresh berries or black pepper berries into value-added white pepper. This work paved way for developing a technological process for microbial decortication of fresh/black pepper for the production of superior-quality white pepper. © 2014 The Society for Applied Microbiology.

  18. Infection of some cayenne pepper varieties (Capsicum frustescens L.) by Tobacco mosaic virus at different growth stages

    Science.gov (United States)

    Damiri, N.; Sofita, I. S.; Effend, T. A.; Rahim, S. E.

    2017-09-01

    This research aimed to study the infection of three varieties of cayenne pepper (Capsicum frustescens L.) by Tobacco Mosaic Virus when they were inoculated at 2, 4, 6, 8 and 10 weeks old after planting. This experiment was conducted in a green house, at the Plant pests and diseases department, Agriculture Faculty, Sriwijaya University, Indralaya, South Sumatra Indonesia from March to October 2014. The study was arranged in factorial completely randomized design with three replicates. First factor was varieties of cayenne pepper namely green, white and small. Second factor was growth stage. Results of the study showed that TMV inoculated at different growth stages of three cayenne pepper varieties affected the incubation period of TMV symptom, time for flowering and productions. The infection of TMV on various ages affected the disease severity on cayenne pepper variety. The highest disease severity was taking place on small cayenne pepper variety that was inoculated at the early stages of age namely 2 weeks after planting. Inoculation of TMV at younger stages of all Cayenne peppers varieties caused a significant reduction in the number of fruits and its weights. TMV has caused a reduction of more than 50% in weight of cayenne pepper fruits regardless of the variety.

  19. An ESR protocol based on relaxation phenomena of irradiated Japanese pepper

    Science.gov (United States)

    Ukai, Mitsuko; Nakamura, Hideo; Shimoyama, Yuhei

    2006-03-01

    We found various free radicals in a commercially available pepper in Japan before and after irradiation using electron spin resonance (ESR) spectroscopy. The typical ESR spectrum of the pepper consists of a sextet centered at g = 2.0, a singlet at the same g-value and a singlet at g = 4.0. Upon gamma ray irradiation, a new pair of signals appeared in the pepper. The progressive saturation behavior (PSB) at various microwave power levels indicated quite different relaxation behaviors of those radicals. Namely, the peak intensity of the organic free radical component decreases in a monotonic fashion, whereas the Mn 2+ and Fe 3+ ESR signals substantially keep constant. This reflects the evidence of three independent radicals in the pepper before irradiation. The PSB of the pair peaks as induced by irradiation possessed quite different PSB from that of the free radical located at g = 2.0. We proposed a new protocol for the ESR detection of irradiated foods by the PSB method at different microwave power levels. This would call for a major modification of the CEN protocol in European Union.

  20. HISTOLOGICAL STUDIES OF THE EFFECTS OF RED PEPPER ON THE STOMACH OF ADULT WISTAR RATS

    Directory of Open Access Journals (Sweden)

    Josiah O. Adjene

    2007-01-01

    Full Text Available Histological effects of red pepper commonly used as spice in food on the stomach of adult wistar rats were carefully investigated. The rats of both sexes (n=24, average weight of 200g were randomly assigned into two treatments (n=16 and control (n=6 groups. The rats in the treatments groups received 1g and 2g of red pepper thoroughly mixed with 20g of their feeds for 7 and 14 days, while the control rats received equal amounts of feeds without the red pepper added. The rats were fed with grower's mash purchased from Edo feeds and flour mill Ltd, Ewu, Edo State and were given water liberally. The rats were sacrificed on day eight and fifteen of the experiment respectively.The stomach was carefully dissected out and quickly fixed in 10% formol saline for routine histological procedure after H & E method.The histological findings after H&E methods indicated that the treated sections of the stomach showed some level of cellular hypertrophy, congestion of blood vessels degenerative changes disruption and distortion of the cytoarchitecture of the stomach.These findings indicate that red pepper may have some deleterious effects on the microanatomy of the stomach of adult wistar rat at higher doses. It is recommended that further studies aimed at corroborating these findings be carried out.

  1. Aroma changes in fresh bell peppers (Capsicum annuum) after hot-air drying.

    NARCIS (Netherlands)

    Luning, P.A.; Yuksel, D.; Vuurst de Vries, van R.; Roozen, J.P.

    1995-01-01

    The aroma of fresh and hot-air dried bell peppers (Capsicum annuum) was evaluated by sensory and instrumental methods. Hot-air drying decreased levels of the odor compounds (Z)-3-hexenal, 2-heptanone, (Z)-2-hexenal, (E)-2-hexenal, hexanol, (Z)-3-hexanol, (E)-2-hexenol, and linalool, which have

  2. Volatile profile and sensory quality of new varieties of Capsicum chinense pepper

    Directory of Open Access Journals (Sweden)

    Deborah dos Santos Garruti

    2013-02-01

    Full Text Available The objective of this study was to compare the sensory quality and the volatile compound profile of new varieties of Capsicum chinense pepper (CNPH 4080 a strain of'Cumari-do-Pará' and BRS Seriema with a known commercial variety (Biquinho. Volatiles were isolated from the headspace of fresh fruit by SPME and identified by GC-MS. Pickled peppers were produced for sensory evaluation. Aroma descriptors were evaluated by Check-All-That-Apply (CATA method, and the frequency data were submitted to Correspondence Analysis. Flavor acceptance was assessed by hedonic scale and analyzed by ANOVA. BRS Seriema showed the richest volatile profile, with 55 identified compounds, and up to 40% were compounds with sweet aroma notes. CNPH 4080 showed similar volatile profile to that of Biquinho pepper, but it had higher amounts of pepper-like and green-note compounds. The samples did not differ in terms of flavor acceptance, but they showed differences in aroma quality confirming the differences found in the volatile profiles. The C. chinense varieties developed by Embrapa proved to be more aromatic than Biquinho variety, and were well accepted by the judges.

  3. Evaluation of pepper spent as an egg yolk colouring agent in the ...

    African Journals Online (AJOL)

    Evaluation of pepper spent as an egg yolk colouring agent in the diet of white leghorn layers. ... Evaluation was based on egg yolk colour intensity measured based on Roche colour fan scores, potassium dichromate grades and consumer preference scores on raw, boiled and fried egg samples. Egg production, feed ...

  4. Effects of moisture content on some physical properties of red pepper

    African Journals Online (AJOL)

    USER

    2010-06-14

    Jun 14, 2010 ... In the moisture range of 7.27 to 20.69% dry basis (d.b.), studies on rewetted red pepper seed ... m-3; ρt, true density, kg m-3; φ, sphericity of seed; al, ... the experiments in this study. ... For each experiment, a sample was.

  5. Anaerobic soil disinfestation reduces survival and infectivity of Phytophthora nicotianae chlamydospores in pepper

    Science.gov (United States)

    Phytophthora nicotianae is the principal causal agent of root and crown rot disease of pepper plants in Extremadura (western Spain), a spring-summer crop in this region. Preplant soil treatment by anaerobic soil disinfestation (ASD) may effectively control plant pathogens in many crop production sys...

  6. Constraints and potential of Sweet Pepper cultivation in plastic houses in Indonesia

    NARCIS (Netherlands)

    Gunadi, N.; Everaarts, A.P.; Adiyoga, W.; Moekasan, T.

    2007-01-01

    At the start of a research project on the improvement of sweet pepper (Capsicum annuum) cultivation in plastic houses in Indonesia, an inventory of the present cultivation methods and constraints was made, together with an analysis of production costs and benefits. The inventory was carried out in

  7. Determination of drying kinetics and sorption isotherm of black pepper (Piper Nigrum)

    Science.gov (United States)

    De Vera, Flordeliza C.; Atienza, Vanessa Bernadette B.; Capili, Jomicah B.; Sauli, Zaliman

    2017-11-01

    In the present study of food products, determination of the drying characteristics of black pepper using an oven is not yet completely established. This study aimed to determine the drying kinetics and sorption isotherm of black pepper using a convective oven at 30°C, 40°C and 50°C. The data gathered in this study were used to fit in selected mathematical models for drying kinetics and sorption isotherm. Among these models, the Midilli model (MR=0.5338exp(0.7273t-0.0551)+-0.0005t for 30°C, MR=0.5814exp(0.6293t-0.0764)+ -0.0008t for 40°C and MR=0.3187exp(1.1777t-0.0466)+ -0.0011t for 50°C) was the best fit to explain the moisture transfer in black pepper, while the GAB Model (m/0.1302=((0.1906)(0.7811)aw)/(1-(0.7811)aw)[1-(0.7811)aw+(0.1906)(0.7811)aw])) was for the equilibrium moisture content and water activity relationship. After evaluating the data, the drying characteristics of black pepper at 40°C yielded better results than 30°C and 50°C. XLSTAT and ANOVA Add-in of Microsoft Excel was the software used to compute for the necessary values in the assessment of the mathematical models for this study.

  8. Improvement of red pepper yield and soil environment by summer catch aquatic crops in greenhouses

    Science.gov (United States)

    Du, X. F.; Wang, L. Z.; Peng, J.; Wang, G. L.; Guo, X. S.; Wen, T. G.; Gu, D. L.; Wang, W. Z.; Wu, C. W.

    2016-08-01

    To investigate effects of the rotation of summer catch crops on remediation retrogressed soils in continuous cropping, a field experiment was conducted. Rice, water spinach, or cress were selected as summer catch crops; bare fallow during summer fallow was used as the control group. Results showed that aquatic crops grown in summer fallow period could effectively reduce soil bulk density and pH, facilitate soil nutrient release, and improve soil physical and chemical properties compared with those grown in fallow period. Paddy-upland rotation could improve soil microbial members and increase bacterial and actinomycete populations; by contrast, paddy-upland rotation could reduce fungal populations and enhance bacterium-to-fungus ratio. Paddy-upland rotation could also actively promote activities of soil enzymes, such as urease, phosphatase, invertase, and catalase. The proposed paddy-upland rotation significantly affected the growth of red pepper; the yield and quality of the grown red pepper were enhanced. Summer catch crops, such as rice, water spinach, and cress significantly increased pepper yield in the following growing season by 15.4%, 10.2% and 14.0%, respectively, compared with those grown in fallow treatment. Therefore, the proposed paddy-upland crop rotation could be a useful method to alleviate continuous cropping problems involved in cultivating red pepper in greenhouses.

  9. De Novo Assembly and Characterization of Fruit Transcriptome in Black Pepper (Piper nigrum).

    Science.gov (United States)

    Hu, Lisong; Hao, Chaoyun; Fan, Rui; Wu, Baoduo; Tan, Lehe; Wu, Huasong

    2015-01-01

    Black pepper is one of the most popular and oldest spices in the world and valued for its pungent constituent alkaloids. Pinerine is the main bioactive compound in pepper alkaloids, which perform unique physiological functions. However, the mechanisms of piperine synthesis are poorly understood. This study is the first to describe the fruit transcriptome of black pepper by sequencing on Illumina HiSeq 2000 platform. A total of 56,281,710 raw reads were obtained and assembled. From these raw reads, 44,061 unigenes with an average length of 1,345 nt were generated. During functional annotation, 40,537 unigenes were annotated in Gene Ontology categories, Kyoto Encyclopedia of Genes and Genomes pathways, Swiss-Prot database, and Nucleotide Collection (NR/NT) database. In addition, 8,196 simple sequence repeats (SSRs) were detected. In a detailed analysis of the transcriptome, housekeeping genes for quantitative polymerase chain reaction internal control, polymorphic SSRs, and lysine/ornithine metabolism-related genes were identified. These results validated the availability of our database. Our study could provide useful data for further research on piperine synthesis in black pepper.

  10. Biting into the Yellow Pepper: The Development of the Interdisciplinary Learner

    Science.gov (United States)

    Haynes, Carolyn

    2014-01-01

    In the opening credits of the television cult classic "Iron Chef," Chairman Kaga takes a huge bite out of a big yellow bell pepper. With the ensuing smirk on his face, he introduces the famous "Kitchen Stadium," which he notes is his "dream in a form never seen before." What follows is a learning experience in which…

  11. Effect of gamma irradiation on the cigarette beetles reared on cayenne pepper

    International Nuclear Information System (INIS)

    Imamura, Taro; Miyanoshita, Akihiro; Todoriki, Setsuko

    2009-01-01

    Effect of gamma irradiation on the survival of the cigarette beetles reared on cayenne pepper was investigated. Gamma ray at a dose of 62 Gy completely killed eggs and larvae of the beetles. Some pupae survived at 540 Gy, but all pupae were killed at 1076 Gy. (author)

  12. Rheoviscometric measurements on starch, flour and black pepper treated by electron-beam and microwaves

    International Nuclear Information System (INIS)

    Ferdes, O.S.; Tirlea, A.; Badea, M.; Casandroiu, T.; Oprita, N.

    1998-01-01

    Irradiation was performed with an ALIN-7 linac within the dose range of 0 to 16 kGy at a dose rate of approximately 1.5 kGy/min. The samples were accommodated in a cavity which was also used for the microwave treatment. The microwave irradiation was carried out inside a special designed system using a 2.45 GHz CW magnetron of 850 W maximum output power. The samples were treated using various power values for irradiation periods varying between 10 and 120 s. Combined treatment by both methods was also applied. The rheoviscometric measurements were carried out on gelified suspensions of starch, flour and black pepper in equivalent starch concentration conditions. For the black pepper suspensions the pH value was adjusted to more than 12 units by adding 33 % NaOH solution. The effects of electromagnetic field and irradiation on starch, flour and pepper were described in terms of apparent viscosity and shear stress. All the samples investigated exhibited similar rheoviscometric behaviour, which can be attributed to starch degradation caused by microwave and electron-beam irradiation, the latter having a more significant effect on the rheological properties. Some consideration were also made concerning the thermal and non-thermal effects of microwaves and of the combined (e-beam and microwave) treatments of starch, flour and pepper

  13. An electron spin resonance study of γ-ray irradiated pepper

    International Nuclear Information System (INIS)

    Ukai, Mitsuko; Hamaya, Naruki; Ichii, Akane; Abe, Aika

    2003-01-01

    Using electron spin resonance (ESR) spectroscopy, we revealed four radical species in the irradiated pepper. The representative ESR spectrum of the pepper is composed of a sextet centered at g=2.0, a singlet at the same g-value, a singlet at g=4.0 and side peaks near g=2.0. The first one is attributable to a signal with hyperfine (hf) interactions of Mn 2+ ion (hf constant=7.4 mT). The second one is due to an organic free radical that may be induced by the (γ-ray irradiation. The third one may be originated from Fe 3+ ion in the non-hem proteins. Those three signals were found in the pepper sample before irradiation. The fourth signals were found at the symmetric position of the organic free radical, i.e., the second signal. The progressive saturation method of the ESR microwave power indicated quite different relaxation behaviors of those radicals. The method reflects four independent radical species in the irradiated pepper. Relaxation time for the singlet signal centered at g=2.0 revealed that the signal is due to the typical organic free radical. (author)

  14. Design and Development of Emittance Measurement Device by Using the Pepper-pot Technique

    Science.gov (United States)

    Pakluea, S.; Rimjaem, S.

    2017-09-01

    Transverse emittance of a charged particle beam is one of the most important properties that reveals the quality of the beam. It is related to charge density, transvers size and angular displacement of the beam in transverse phase space. There are several techniques to measure the transverse emittance value. One of practical methods is the pepper-pot technique, which can measure both horizontal and vertical emittance value in a single measurement. This research concentrates on development of a pepper-pot device to measure the transverse emittance of electron beam produced from an accelerator injector system, which consists of a thermionic cathode RF electron gun and an alpha magnet, at the Plasma and Beam Physics Research Facility, Chiang Mai University. Simulation of beam dynamics was conducted with programs PARMELA, ELEGANT and self-developed codes using C and MATLAB. The geometry, dimensions and location of the pepper-pot as well as its corresponding screen station position were included in the simulation. The result from this study will be used to design and develop a practical pepper-pot experimental station.

  15. Enzymatic firming of processed red pepper by means of exogenous pectinesterase

    DEFF Research Database (Denmark)

    Jensen, M.; Petersen, B.R.; Adler-Nissen, Jens

    2004-01-01

    The objective of this investigation was to demonstrate that the firmness of a commercial vegetable product, diced and frozen red pepper (Capsicum annum var. Sendt), could be improved by the use of exogenous pectinesterase in an industrially relevant process. The diced pepper pieces 10 x 10 x 7 mm(3......) were infused under vacuum with a commercially available pectinesterase. The range of optimal process conditions was: 15-20degrees C, 45 min infusion time, a 10-25 mM CaCl2 infusion brine, a w/w ratio of pepper fruit to infusion brine of 1.5:1, and an enzyme dosage of 30-60 pectinesterase units (PEU......) per kg pepper fruit. The firmness as measured by back extrusion was improved by a factor of two to three. The effect of firming was robust and conserved after freezing and heating in a simulated household cooking process. The firming process seems easily adaptable to industrial conditions and may...

  16. Factors affecting oviposition of Bemisia tabaci (Genn.) biotype B (Hemiptera: Aleyrodidae) in sweet pepper

    International Nuclear Information System (INIS)

    Lima, Larissa C. de; Campos, Alcebiades R.

    2008-01-01

    Bemisia tabaci (Gen.) biotype B is considered a pest of economical importance for several vegetables. The oviposition behaviour of the while fly was evaluated in sweet pepper plants. The trials were carried out under greenhouse condition and in the Laboratory of Entomology of DEFERS/ UNESP, Campus of Ilha Solteira-SP, with the sweet pepper Magali-R genotype. The effect of plant age on the whitefly oviposition was evaluated in free-choice tests, in plants, 25, 30, 35, 40 and 45- day-old, as egg distribution in the plant and on the leaf blade was evaluated in 35-days-old plants. In a no-choice tests, 35-day-old plants were used to evaluate the effect of the densities of 50, 100, 150, 200 and 250 adults per plant on the number of eggs laid by insects. The silver leaf whitefly preferred to oviposition the third to sixth leaflets, of the medium and superior part of plants of sweet pepper; the leaf blade areas, located in the lobes right and left close the base of the leaf were the preferential site for whitefly oviposition. Older plants, 40- and 45-day-old, were preferentially used for oviposition, and 200 and 250 adults per plant were both enough to lay a number of eggs that allowed to differentiate among sweet pepper genotypes with different whitefly resistance levels. (author)

  17. Burkholderia sp. KCTC 11096BP modulates pepper growth and resistance against Phytophthora capsici

    International Nuclear Information System (INIS)

    Kang, S.M.; Hamayun, M.; Shinwari, Z.K.

    2016-01-01

    Biological control of crop diseases is desirable for sustainable agriculture as it minimizes chemical inputs in the agricultural system and promotes eco-friendly environment. We analyzed the favorable role of Burkholderia sp. KCTC 11096BP against the pathogen Phytophthora capsici in pepper. We screen thirty rhizobateria for their anti-pathogen activity, and found that Burkholderia sp. KCTC 11096BP exhibits maximum growth inhibition of the pathogen P. capsici. The bacterium inoculation to pepper plants significantly enhanced growth attributes of pepper in infected and control treatments. The total proteins (10.9%), and the amino acids viz. glycine (4.08 ug/g), leucine (3.3 ug/g), and alanine (3.26 ug/g) were preset in considerably higher quantities in Burkholderia sp. applied treatments as compare to control. The systemic acquired resistance (SAR) of the host plant was up-regulated by Burkholderia sp. KCTC, as endogenous salicylic acid (235.5 ng/g) and jasmonic acid (22.8 ng/g) levels were found higher in such treatments. It was concluded that Burkholderia sp. KCTC 11096BP mitigates the adverse effects of P. capsici on pepper crop and can improve crop productivity at the field level. (author)

  18. Development of Colletotrichum gloeosporioides isolated from green pepper in different culture media, temperatures, and light regimes

    Directory of Open Access Journals (Sweden)

    Mello Alexandre Furtado Silveira

    2004-01-01

    Full Text Available Control of anthracnose in green pepper involves the use of resistant varieties and/or fungicides. The selection of varieties and efficient products demands great amounts of conidia as inoculum. It is thus necessary to optimize the production of Colletotrichum gloeosporioides conidia in the laboratory, establishing the best conditions for fungus development. The present study aimed at determining the most favorable culture media, temperature, and light conditions for the production of fungus inoculum. The fungus was isolated from green pepper fruits (Capsicum annuum L. and transferred to four culture media (PDA, oat, filtered pepper extract, and autoclaved pepper extract, under different temperatures (15, 20, 25, 30, and 35ºC and light conditions (24h dark, and 24h light. Colony growth was evaluated after 7 and 12 days of incubation. No differences were found between the culture media. However, the greatest number of conidia was obtained from colonies grown in oat medium at 25ºC. Temperatures of 20 and 25ºC were the most favorable for colony growth and sporulation. Higher sporulation was obtained under incubation in constant light. Cultivation of C. gloeosporioides in oat medium, at 25ºC, and constant light is recommended.

  19. An investigation of a defensive chitinase against Fusarium oxysporum in pepper leaf tissue

    Directory of Open Access Journals (Sweden)

    Khemika S. Lomthaisong

    2008-01-01

    Full Text Available Plant chitinase is classified as a PR-protein involved in a defense mechanism against a pathogen. This research aims to investigate a specific type of chitinase which is produced by pepper in response to an early defense against Fusarium oxysporum, which causes wilt disease. The changes of chitinase isozyme patterns in the inter- and intracellular fluids in the leaf of four cultivars of pepper (Capsicum annuum L. at day 1, 3, 5, 7 and 10 from fungal inoculation were analysed using SDS-PAGE in polyacrylamide gel supplemented with glycol chitin as a substrate. The levels of disease severity in the four varieties of pepper were also compared with the isozyme patterns. The results showed that the resistance of pepper to F. oxysporum attack corresponded to the expression of ~70 kDa chitinase band (Chi-3 in the intercellular fluid. Therefore, such chitinase could possibly be used as a protein marker to identify the tolerant line and as a springboard for further study of wilt disease control.

  20. Toward an optimal control strategy for sweet pepper cultivation : a dynamic crop model

    NARCIS (Netherlands)

    Buwalda, F.; Henten, van E.J.; Gelder, de A.; Bontsema, J.; Hemming, J.

    2006-01-01

    Sweet pepper production is characterized by large fluctuations in fruit yield per week. Synchronization of yield patterns between nurseries in response to weather conditions leads to variations in market supply and affects price formation. In order to improve supply chain efficiency, auctions and

  1. Chitosan controls postharvest anthracnose in bell pepper by activating defense-related enzymes.

    Science.gov (United States)

    Edirisinghe, Madushani; Ali, Asgar; Maqbool, Mehdi; Alderson, Peter G

    2014-12-01

    Anthracnose, a postharvest disease caused by the fungus Colletotrichum capsici is the most devastating disease of bell pepper that causes great economic losses especially in tropical climates. Therefore, the objective of this study was to evaluate the antifungal properties of chitosan (low molecular weight from crab shell, Mw: 50 kDa and 75-85 % deacetylated) against anthracnose by inducing defense-related enzymes. The concentrations of 0, 0.5, 1.0, 1.5 and 2.0 % chitosan were used to control the fungus in vitro and postharvest. There was a reduction in C. capsici mycelial growth and the highest chitosan concentration (2.0 %) reduced the growth by 70 % after 7 days incubation. In germination test, the concentration of 1.5 and 2.0 % chitosan reduced spore germination in C. capsici between 80 % and 84 %, respectively. In postharvest trial the concentration of 1.5 % decreased the anthracnose severity in pepper fruit by approximately 76 % after 28 days of storage (10 ± 1 °C; 80 % RH). For enzymatic activities, the concentration of 1.5 and 2.0 % chitosan increased the polyphenol oxidase (PPO), peroxidase (POD) and total phenolics in inoculated bell pepper during storage. Based on these results, the chitosan presents antifungal properties against C. capsici, as well as potential to induce resistance on bell pepper.

  2. Application of Volatile Antifungal Plant Essential Oils for Controlling Pepper Fruit Anthracnose by Colletotrichum gloeosporioides

    Directory of Open Access Journals (Sweden)

    Jeum Kyu Hong

    2015-09-01

    Full Text Available Anthracnose caused by Colletotrichum gloeosporioides has been destructive during pepper fruit production in outdoor fields in Korea. In vitro antifungal activities of 15 different plant essential oils or its components were evaluated during conidial germination and mycelial growth of C. gloeosporioides. In vitro conidial germination was most drastically inhibited by vapour treatments with carvacrol, cinnamon oil, trans-cinnamaldehyde, citral, p-cymene and linalool. Inhibition of the mycelial growth by indirect vapour treatment with essential oils was also demonstrated compared with untreated control. Carvacrol, cinnamon oil, trans-cinnamaldehyde, citral and eugenol were among the most inhibitory plant essential oils by the indirect antifungal efficacies. Plant protection efficacies of the plant essential oils were demonstrated by reduced lesion diameter on the C. gloeosporioides-inoculated immature green pepper fruits compared to the inoculated control fruits without any plant essential oil treatment. In planta test showed that all plant essential oils tested in this study demonstrated plant protection efficacies against pepper fruit anthracnose with similar levels. Thus, application of different plant essential oils can be used for eco-friendly disease management of anthracnose during pepper fruit production.

  3. Application of Volatile Antifungal Plant Essential Oils for Controlling Pepper Fruit Anthracnose by Colletotrichum gloeosporioides.

    Science.gov (United States)

    Hong, Jeum Kyu; Yang, Hye Ji; Jung, Heesoo; Yoon, Dong June; Sang, Mee Kyung; Jeun, Yong-Chull

    2015-09-01

    Anthracnose caused by Colletotrichum gloeosporioides has been destructive during pepper fruit production in outdoor fields in Korea. In vitro antifungal activities of 15 different plant essential oils or its components were evaluated during conidial germination and mycelial growth of C. gloeosporioides. In vitro conidial germination was most drastically inhibited by vapour treatments with carvacrol, cinnamon oil, trans-cinnamaldehyde, citral, p-cymene and linalool. Inhibition of the mycelial growth by indirect vapour treatment with essential oils was also demonstrated compared with untreated control. Carvacrol, cinnamon oil, trans-cinnamaldehyde, citral and eugenol were among the most inhibitory plant essential oils by the indirect antifungal efficacies. Plant protection efficacies of the plant essential oils were demonstrated by reduced lesion diameter on the C. gloeosporioides-inoculated immature green pepper fruits compared to the inoculated control fruits without any plant essential oil treatment. In planta test showed that all plant essential oils tested in this study demonstrated plant protection efficacies against pepper fruit anthracnose with similar levels. Thus, application of different plant essential oils can be used for eco-friendly disease management of anthracnose during pepper fruit production.

  4. Abortion of reproductive organs in sweet pepper (Capsicum annuum L.): a review

    NARCIS (Netherlands)

    Wubs, A.M.; Heuvelink, E.; Marcelis, L.F.M.

    2009-01-01

    Levels of abortion of reproductive organs (i.e., buds, flowers, and young fruits) in sweet pepper plants (Capsicum annuum L.) are high, and cyclical fluctuations occur in fruit set. Stages susceptible to abortion are very young buds (<2.5 mm), buds close to anthesis, and flowers and fruits up to

  5. Employing airborne multispectral digital imagery to map Brazilian pepper infestation in south Texas.

    Science.gov (United States)

    A study was conducted in south Texas to determine the feasibility of using airborne multispectral digital imagery for differentiating the invasive plant Brazilian pepper (Schinus terebinthifolius) from other cover types. Imagery obtained in the visible, near infrared, and mid infrared regions of th...

  6. Structure and genetic diversity of natural Brazilian pepper populations (Schinus terebinthifolius Raddi).

    Science.gov (United States)

    Álvares-Carvalho, S V; Duarte, J F; Santos, T C; Santos, R M; Silva-Mann, R; Carvalho, D

    2016-06-17

    In the face of a possible loss of genetic diversity in plants due the environmental changes, actions to ensure the genetic variability are an urgent necessity. The extraction of Brazilian pepper fruits is a cause of concern because it results in the lack of seeds in soil, hindering its distribution in space and time. It is important to address this concern and explore the species, used by riparian communities and agro-factories without considering the need for keeping the seeds for natural seed banks and for species sustainability. The objective of this study was to evaluate the structure and the genetic diversity in natural Brazilian pepper populations (Schinus terebinthifolius Raddi). Twenty-two alleles in 223 individuals were identified from eight forest remnants located in the states of Minas Gerais, Espírito Santo, and Sergipe. All populations presented loci in Hardy-Weinberg equilibrium deviation. Four populations presented six combinations of loci in linkage disequilibrium. Six exclusive alleles were detected in four populations. Analysis of molecular variance showed the absence of diversity between regions and that between the populations (GST) was 41%. Genetic diversity was structured in seven clusters (ΔK7). Brazilian pepper populations were not structured in a pattern of isolation by distance and present genetic bottleneck. The populations São Mateus, Canastra, Barbacena, and Ilha das Flores were identified as management units and may support conservation projects, ecological restoration and in implementation of management plans for Brazilian pepper in the State of Sergipe.

  7. Foliar diagnosis, nutrition and yield stability of black pepper (Piper nigrum L.) in Sarawak

    NARCIS (Netherlands)

    Waard, de P.W.F.

    1969-01-01

    Until 1942 cultivation of pepper P. nigrum L. in Sarawak produced relatively small but regular yields. High demands after 1945 and restricted use of "burnt earth" compelled farmers to abandon the application of this traditional fertilizer. Instead, "fool proof" manufactured fertilizers

  8. Cytokinin response in pepper plants (Capsicum annuum L.) exposed to silver nanoparticles

    Czech Academy of Sciences Publication Activity Database

    Vinković, T.; Novák, Ondřej; Strnad, Miroslav; Goessler, W.; Jurašin, D. D.; Parađiković, N.; Vrček, I. V.

    2017-01-01

    Roč. 156, JUL (2017), s. 10-18 ISSN 0013-9351 R&D Projects: GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : Abiotic stress * Biodistribution * Cytokinin response * Pepper * Silver nanoparticles * Uptake Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 3.835, year: 2016

  9. The evolution of chili peppers (Capsicum-Solanaceae): a cytogenetic perspective

    Science.gov (United States)

    Capsicum (chili peppers) is a New World genus with five crop species of great economic importance for food and spices. An up-to-date summary of the karyotypic knowledge is presented, including data on classical staining (chromosome number, size and morphology), silver impregnation (number and positi...

  10. Gamma radiation effect on Bacillus cereus spores inoculated in black pepper

    International Nuclear Information System (INIS)

    Froehlich, Angela; Axeredo, Raquel M.C.; Vanetti, Maria Cristina D.

    2000-01-01

    It had been analyzed 37 samples of worn out black pepper and in 85% of these samples was observed the presence of Bacillus cereus in numbers of up to 4,6 x 10 4 UFC/g. The population of aerobic mesofilis bacteria varied of 2,8 x 10 5 the 1,9 x 10 8 UFC/g. The black pepper used during the experiment was evaluated, evidencing the aerobic presence of one aerobic mesofilis microbiota of, approximately, 2,6 x 10 6 UFC/g, consisting, mainly, for species of the Bacillus sort. It was observed that the absence of B. cereus, coliforms, filamentous fungus and leavenings. The evaluation of the irradiation of the black pepper inoculated with 10 6 UFC/g of B. cereus spores of with doses of gamma radiation varying between 2 and 10 kGy evidenced that doses up to 5 kGy had been enough to reduce the counting of, approximately, 10 6 UFC/g of aerobic mesofilis organisms and 10 4 UFC/g of B. cereus spores the not detectable numbers by the used methodology. The dose of reduction decimal (D 10 ) for the inoculated B. cereus spores in black pepper was of 1,78 kGy

  11. Interactions of Phytophthora capsici with Resistant and Susceptible Pepper Roots and Stems.

    Science.gov (United States)

    Dunn, Amara R; Smart, Christine D

    2015-10-01

    Using host resistance is an important strategy for managing pepper root and crown rot caused by Phytophthora capsici. An isolate of P. capsici constitutively expressing a gene for green fluorescent protein was used to investigate pathogen interactions with roots, crowns, and stems of Phytophthora-susceptible bell pepper 'Red Knight', Phytophthora-resistant bell pepper 'Paladin', and Phytophthora-resistant landrace Criollos de Morelos 334 (CM-334). In this study, the same number of zoospores attached to and germinated on roots of all cultivars 30 and 120 min postinoculation (pi), respectively. At 3 days pi, significantly more secondary roots had necrotic lesions on Red Knight than on Paladin and CM-334 plants. By 4 days pi, necrotic lesions had formed on the taproot of Red Knight but not Paladin or CM-334 plants. Although hyphae were visible in the crowns and stems of all Red Knight plants observed at 4 days pi, hyphae were observed in crowns of only a few Paladin and in no CM-334 plants, and never in stems of either resistant cultivar at 4 days pi. These results improve our understanding of how P. capsici infects plants and may contribute to the use of resistant pepper cultivars for disease management and the development of new cultivars.

  12. Effectiveness and safety of botanical pesticides applied in black pepper (Piper nigrum) plantations

    NARCIS (Netherlands)

    Wiratno,

    2008-01-01

    Black pepper (Piper nigrum L) is an important commodity of Indonesia, which has been cultivated since the 6th century. The plant plays an important role in local economies since 95% of the plantations are cultivated by smallholder farmers. Because of this important economic value, proper plant

  13. Surveys of virus diseases on pepper ( Capsicum spp.) in South-west ...

    African Journals Online (AJOL)

    Surveys to determine the incidence, diversity and distribution of viruses infecting pepper (Capsicum spp.) were conducted in six states (Oyo, Ondo, Osun, Ogun, Ekiti and Lagos) of South-west Nigeria in 2010 and 2011. Leaf samples from symptomatic and asymptomatic plants were collected at random from farmers' fields ...

  14. Consumer and farmer safety evaluation of application of botanical pesticides in black pepper crop protection

    NARCIS (Netherlands)

    Hernandez-Moreno, J.; Soffers, A.E.M.F.; Wiratno,; Falke, H.E.; Rietjens, I.; Murk, A.J.

    2013-01-01

    This study presents a consumer and farmer safety evaluation on the use of four botanical pesticides in pepper berry crop protection. The pesticides evaluated include preparations from clove, tuba root, sweet flag and pyrethrum. Their safety evaluation was based on their active ingredients being

  15. Innovations in microspore embryogenesis in Indonesian hot pepper (Capsicum annuum L.) and Brassica napus L.

    NARCIS (Netherlands)

    Supena, E.D.J.

    2004-01-01

    Hot pepper (Capsicum annuumL.) is the most important vegetable in

  16. Phylogenetic relationships, diversification and expansion of chili peppers (Capsicum, Solanaceae).

    Science.gov (United States)

    Carrizo García, Carolina; Barfuss, Michael H J; Sehr, Eva M; Barboza, Gloria E; Samuel, Rosabelle; Moscone, Eduardo A; Ehrendorfer, Friedrich

    2016-07-01

    Capsicum (Solanaceae), native to the tropical and temperate Americas, comprises the well-known sweet and hot chili peppers and several wild species. So far, only partial taxonomic and phylogenetic analyses have been done for the genus. Here, the phylogenetic relationships between nearly all taxa of Capsicum were explored to test the monophyly of the genus and to obtain a better knowledge of species relationships, diversification and expansion. Thirty-four of approximately 35 Capsicum species were sampled. Maximum parsimony and Bayesian inference analyses were performed using two plastid markers (matK and psbA-trnH) and one single-copy nuclear gene (waxy). The evolutionary changes of nine key features were reconstructed following the parsimony ancestral states method. Ancestral areas were reconstructed through a Bayesian Markov chain Monte Carlo analysis. Capsicum forms a monophyletic clade, with Lycianthes as a sister group, following both phylogenetic approaches. Eleven well-supported clades (four of them monotypic) can be recognized within Capsicum, although some interspecific relationships need further analysis. A few features are useful to characterize different clades (e.g. fruit anatomy, chromosome base number), whereas some others are highly homoplastic (e.g. seed colour). The origin of Capsicum is postulated in an area along the Andes of western to north-western South America. The expansion of the genus has followed a clockwise direction around the Amazon basin, towards central and south-eastern Brazil, then back to western South America, and finally northwards to Central America. New insights are provided regarding interspecific relationships, character evolution, and geographical origin and expansion of Capsicum A clearly distinct early-diverging clade can be distinguished, centred in western-north-western South America. Subsequent rapid speciation has led to the origin of the remaining clades. The diversification of Capsicum has culminated in the origin

  17. Age-related and sex-specific differences in proteasome activity in individual Drosophila flies from wild type, longevity-selected and stress resistant strains

    DEFF Research Database (Denmark)

    Hansen, Tina Østergaard; Sarup, Pernille Merete; Loeschcke, Volker

    2012-01-01

    with that in C1 males. However, in longevity-selected LS1 flies the proteasome activity was significantly lower compared to C1 flies, but the sex differences were maintained to some extent. Five other stress resistant lines also had significantly reduced proteasome activity in both sexes. During ageing...... and that increased lifespan and stress resistance lead to a reduction in proteasome activity and recession of the age-related decline observed in control females....

  18. Aconitase B is required for optimal growth of Xanthomonas campestris pv. vesicatoria in pepper plants.

    Directory of Open Access Journals (Sweden)

    Janine Kirchberg

    Full Text Available The aerobic plant pathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv colonizes the intercellular spaces of pepper and tomato. One enzyme that might contribute to the successful proliferation of Xcv in the host is the iron-sulfur protein aconitase, which catalyzes the conversion of citrate to isocitrate in the tricarboxylic acid (TCA cycle and might also sense reactive oxygen species (ROS and changes in cellular iron levels. Xcv contains three putative aconitases, two of which, acnA and acnB, are encoded by a single chromosomal locus. The focus of this study is aconitase B (AcnB. acnB is co-transcribed with two genes, XCV1925 and XCV1926, encoding putative nucleic acid-binding proteins. In vitro growth of acnB mutants was like wild type, whereas in planta growth and symptom formation in pepper plants were impaired. While acnA, XCV1925 or XCV1926 mutants showed a wild-type phenotype with respect to bacterial growth and in planta symptom formation, proliferation of the acnB mutant in susceptible pepper plants was significantly impaired. Furthermore, the deletion of acnB led to reduced HR induction in resistant pepper plants and an increased susceptibility to the superoxide-generating compound menadione. As AcnB complemented the growth deficiency of an Escherichia coli aconitase mutant, it is likely to be an active aconitase. We therefore propose that optimal growth and survival of Xcv in pepper plants depends on AcnB, which might be required for the utilization of citrate as carbon source and could also help protect the bacterium against oxidative stress.

  19. Using Of Viscosity Property For Identification Of Irradiated Black Pepper And Cumin

    International Nuclear Information System (INIS)

    ALI, H.G.M.

    2009-01-01

    In the present investigation, the viscosity measurement was used for detecting irradiated black pepper and cumin during storage period. All samples under investigation were packed in polyethylene bags then irradiated at 3, 5, 7 and 10 kGy and stored for 12 months at room temperature then viscosity were measured under extremely alkaline condition (ph 13.8). The results indicated that irradiation treatment caused significant decrease in apparent viscosity values, where it decreased from 67.3 in control sample to 49.7, 42.3, 38.7 and 32.7 mpa.s in black pepper, while the viscosity of cumin was decreased from 74 mpa.s for control to 64.9, 41.7, 25.3 and 11.3 mpa.s for samples. The viscosity of black pepper and cumin were decreased significantly with increasing the radiation dose and during storage period. It was decreased from 67.3 for control to 67.0, 47.7, 32.3 and 17.7 mpa.s after 3, 6, 9 and 12 months, respectively, in black pepper while the viscosity of cumin samples was decreased significantly from 74 for control to 50.3, 17.1, 9.5 and 7.2 mpa.s after the same months. The data also showed that the moisture and carbohydrate contents were decreased by increasing the radiation dose. It could be concluded that the viscosity parameter can be used to identify of black pepper and cumin samples after irradiation and during 12 months of storage at ambient temperature.

  20. Arbuscular mycorrhizal fungi (AMF) as bio protector agents against wilt induced by Verticillium spp. in pepper

    Energy Technology Data Exchange (ETDEWEB)

    Goicoechea, N.; Garmendia, I.; Sanchez-Diaz, M.; Aguirreolea, J.

    2010-07-01

    Verticillium dahliae Kleb. is a vascular pathogen that alters water status and growth of pepper plants and causes drastic reductions in yield. Its control is difficult because it can survive in field soil for several years. The application of arbuscular mycorrhizal fungi (AMF) as bio protector agents against V. dahliae is an alternative to the use of chemicals which, in addition, is more respectful with the environment. The establishment of the mutualistic association of plant roots and AMF involves a continuous cellular and molecular dialogue between both symbionts that includes the pre activation of plant defense responses that may enhance the resistance or tolerance of mycorrhizal plants to soil-borne pathogens. Some AMF can improve the resistance of Capsicum annuum L. against V. dahliae. This is especially relevant for pepper cultivars (i.e. cv. Piquillo) that exhibit high susceptibility to this pathogen. Compared with non-mycorrhizal plants, mycorrhizal pepper can exhibit more balanced antioxidant metabolism in leaves along the first month after pathogen inoculation, which may contribute to delay both the development of disease symptoms and the decrease of photosynthesis in Verticillium-inoculated plants with the subsequent benefit for yield. In stems, mycorrhizal pepper show earlier and higher deposition of lignin in xylem vessels than non mycorrhizal plants, even in absence of the pathogen. Moreover, AMF can induce new isoforms of acidic chitinases and superoxide dismutase in roots. Mycorrhizal-specific induction of these enzymatic activities together with enhanced peroxidase and phenylalanine ammonia-lyase in roots may also be involved in the bio protection of Verticillium-induced wilt in pepper by AMF. (Author) 81 refs.