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Sample records for citrus hsp70 molecular

  1. Identification and in silico analysis of the Citrus HSP70 molecular chaperone gene family

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    Luciano G. Fietto

    2007-01-01

    Full Text Available The completion of the genome sequencing of the Arabidopsis thaliana model system provided a powerful molecular tool for comparative analysis of gene families present in the genome of economically relevant plant species. In this investigation, we used the sequences of the Arabidopsis Hsp70 gene family to identify and annotate the Citrus Hsp70 genes represented in the CitEST database. Based on sequence comparison analysis, we identified 18 clusters that were further divided into 5 subgroups encoding four mitochondrial mtHsp70s, three plastid csHsp70s, one ER luminal Hsp70 BiP, two HSP110/SSE-related proteins and eight cytosolic Hsp/Hsc70s. We also analyzed the expression profile by digital Northern of each Hsp70 transcript in different organs and in response to stress conditions. The EST database revealed a distinct population distribution of Hsp70 ESTs among isoforms and across the organs surveyed. The Hsp70-5 isoform was highly expressed in seeds, whereas BiP, mitochondrial and plastid HSp70 mRNAs displayed a similar expression profile in the organs analyzed, and were predominantly represented in flowers. Distinct Hsp70 mRNAs were also differentially expressed during Xylella infection and Citrus tristeza viral infection as well as during water deficit. This in silico study sets the groundwork for future investigations to fully characterize functionally the Citrus Hsp70 family and underscores the relevance of Hsp70s in response to abiotic and biotic stresses in Citrus.

  2. Identification and in silico analysis of the Citrus HSP70 molecular chaperone gene family

    OpenAIRE

    Fietto, Luciano G.; Maximiller D.L. Costa; Cosme D Cruz; De Souza, Alessandra A.; Machado, Marcos A; Fontes, Elizabeth P. B.

    2007-01-01

    The completion of the genome sequencing of the Arabidopsis thaliana model system provided a powerful molecular tool for comparative analysis of gene families present in the genome of economically relevant plant species. In this investigation, we used the sequences of the Arabidopsis Hsp70 gene family to identify and annotate the Citrus Hsp70 genes represented in the CitEST database. Based on sequence comparison analysis, we identified 18 clusters that were further divided into 5 subgroups enc...

  3. The nucleotide exchange factors of Hsp70 molecular chaperone

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    Andreas eBracher

    2015-04-01

    Full Text Available Molecular chaperones of the Hsp70 family form an important hub in the cellular protein folding networks in bacteria and eukaryotes, connecting translation with the downstream machineries of protein folding and degradation. The Hsp70 folding cycle is driven by two types of cochaperones: J-domain proteins stimulate ATP hydrolysis by Hsp70, while nucleotide exchange factors (NEFs promote replacement of Hsp70-bound ADP with ATP. Bacteria and organelles of bacterial origin have only one known NEF type for Hsp70, GrpE. In contrast, a large diversity of Hsp70 NEFs has been discovered in the eukaryotic cell. These NEFs belong to the Hsp110/Grp170, HspBP1/Sil1 and BAG domain protein families. In this short review we compare the structures and molecular mechanisms of nucleotide exchange factors for Hsp70 and discuss how these cochaperones contribute to protein folding and quality control in the cell.

  4. Zinc-L-carnosine binds to molecular chaperone HSP70 and inhibits the chaperone activity of the protein.

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    Haga, Asami; Okamoto, Tomoya; Yamada, Shintaroh; Kubota, Toshihiko; Sanpei, Ann; Takahashi, Shota; Nakayama, Masahiro; Nagai, Miki; Otaka, Michiro; Miyazaki, Toshio; Nunomura, Wataru; Grave, Ewa; Itoh, Hideaki

    2013-09-01

    In this study, we have investigated the specific binding proteins of Zinc-L-carnosine (Polaprezinc) using Polaprezinc-affinity column chromatography in vitro. A protein having a 70-kDa molecular mass was eluted by the linear gradient of 0-1.0 mM Polaprezinc from the affinity column and the protein was identified as the molecular chaperone HSP70 by immunoblotting. The chaperone activity of HSP70 was completely suppressed by Polaprezinc. The ATPase activity of HSP70 was affected to some extent by the reagent. In the circular dichroism (CD) spectrum, the secondary structure of HSP70 was changed in the presence of Polaprezinc, i.e. it decreased in the α-helix. We have determined the Polaprezinc-binding domain of HSP70 by using recombinant HSP70N- and C-domains. Although Polaprezinc could bind to both the N-terminal and the C-terminal of HSP70, the HSP70N-domain has a high affinity to the drug. Regarding the peptide cleavage of the HSP70N- and C-domains with proteinase K, the intact HSP70N still remained in the presence of Polaprezinc. On the other hand, the quantity of the intact C-domain slightly decreased under the same conditions along with the newly digested small peptides appeared. It has been suggested that Polaprezinc binds to HSP70 especially in the N-domains, suppresses the chaperone activity and delays an ATPase activities of HSP70. PMID:23687308

  5. Molecular mechanism of mutant p53 stabilization: the role of HSP70 and MDM2.

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    Milena Wiech

    Full Text Available Numerous p53 missense mutations possess gain-of-function activities. Studies in mouse models have demonstrated that the stabilization of p53 R172H (R175H in human mutant protein, by currently unknown factors, is a prerequisite for its oncogenic gain-of-function phenotype such as tumour progression and metastasis. Here we show that MDM2-dependent ubiquitination and degradation of p53 R175H mutant protein in mouse embryonic fibroblasts is partially inhibited by increasing concentration of heat shock protein 70 (HSP70/HSPA1-A. These phenomena correlate well with the appearance of HSP70-dependent folding intermediates in the form of dynamic cytoplasmic spots containing aggregate-prone p53 R175H and several molecular chaperones. We propose that a transient but recurrent interaction with HSP70 may lead to an increase in mutant p53 protein half-life. In the presence of MDM2 these pseudoaggregates can form stable amyloid-like structures, which occasionally merge into an aggresome. Interestingly, formation of folding intermediates is not observed in the presence of HSC70/HSPA8, the dominant-negative K71S variant of HSP70 or HSP70 inhibitor. In cancer cells, where endogenous HSP70 levels are already elevated, mutant p53 protein forms nuclear aggregates without the addition of exogenous HSP70. Aggregates containing p53 are also visible under conditions where p53 is partially unfolded: 37°C for temperature-sensitive variant p53 V143A and 42°C for wild-type p53. Refolding kinetics of p53 indicate that HSP70 causes transient exposure of p53 aggregate-prone domain(s. We propose that formation of HSP70- and MDM2-dependent protein coaggregates in tumours with high levels of these two proteins could be one of the mechanisms by which mutant p53 is stabilized. Moreover, sequestration of p73 tumour suppressor protein by these nuclear aggregates may lead to gain-of-function phenotypes.

  6. Genetic variation and recombination of RdRp and HSP 70h genes of Citrus tristeza virus isolates from orange trees showing symptoms of citrus sudden death disease

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    Pappas Georgios J

    2008-01-01

    Full Text Available Abstract Background Citrus sudden death (CSD, a disease that rapidly kills orange trees, is an emerging threat to the Brazilian citrus industry. Although the causal agent of CSD has not been definitively determined, based on the disease's distribution and symptomatology it is suspected that the agent may be a new strain of Citrus tristeza virus (CTV. CTV genetic variation was therefore assessed in two Brazilian orange trees displaying CSD symptoms and a third with more conventional CTV symptoms. Results A total of 286 RNA-dependent-RNA polymerase (RdRp and 284 heat shock protein 70 homolog (HSP70h gene fragments were determined for CTV variants infecting the three trees. It was discovered that, despite differences in symptomatology, the trees were all apparently coinfected with similar populations of divergent CTV variants. While mixed CTV infections are common, the genetic distance between the most divergent population members observed (24.1% for RdRp and 11.0% for HSP70h was far greater than that in previously described mixed infections. Recombinants of five distinct RdRp lineages and three distinct HSP70h lineages were easily detectable but respectively accounted for only 5.9 and 11.9% of the RdRp and HSP70h gene fragments analysed and there was no evidence of an association between particular recombinant mosaics and CSD. Also, comparisons of CTV population structures indicated that the two most similar CTV populations were those of one of the trees with CSD and the tree without CSD. Conclusion We suggest that if CTV is the causal agent of CSD, it is most likely a subtle feature of population structures within mixed infections and not merely the presence (or absence of a single CTV variant within these populations that triggers the disease.

  7. Molecular cloning of mtp70, a mitochondrial member of the hsp70 family.

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    Engman, D M; Kirchhoff, L. V.; Donelson, J E

    1989-01-01

    We have isolated a gene from the protozoan parasite Trypanosoma cruzi that encodes a previously unidentified member of the 70-kilodalton heat shock protein (hsp70) family. Among all the eucaryotic hsp70 proteins described to date, this trypanosome protein, mtp70, is uniquely related in sequence and structure to the hsp70 of Escherichia coli, DnaK, which functions in the initiation of DNA replication. This relationship to DnaK is especially relevant in view of the intracellular location of the...

  8. Insights into the molecular mechanism of allostery in Hsp70s

    Science.gov (United States)

    Mayer, Matthias P.; Kityk, Roman

    2015-01-01

    Hsp70s chaperone an amazing number and variety of cellular protein folding processes. Key to their versatility is the recognition of a short degenerate sequence motif, present in practically all polypeptides, and a bidirectional allosteric intramolecular regulation mechanism linking their N-terminal nucleotide binding domain (NBD) and their C-terminal polypeptide substrate binding domain (SBD). Through this interdomain communication ATP binding to the NBD and ATP hydrolysis control the affinity of the SBD for polypeptide substrates and substrate binding to the SBD triggers ATP hydrolysis. Genetic screens for defective variants of Hsp70s and systematic analysis of available structures of the isolated domains revealed some residues involved in allosteric control. Recent elucidation of the crystal structure of the Hsp70 homolog DnaK in the ATP bound open conformation as well as numerous NMR and mutagenesis studies bring us closer to an understanding of the communication between NBD and SBD. In this review we will discuss our current view of the allosteric control mechanism of Hsp70 chaperones. PMID:26539440

  9. Molecular characterization and expression of HSP70, HSF and HSBP genes inOctopus vulgaris during thermal stress

    Institute of Scientific and Technical Information of China (English)

    HONG Jingni; MAO Yong; NIU Sufang; SUN Tiantian; SU Yongquan

    2015-01-01

    Temperature is an important environmental factor that affects the growth and survival ofOctopus vulgaris, the common octopus. To understand the protective mechanism thatO.vulgaris exhibits under heat stress, we used rapid amplification of cDNA ends (RACE) to obtain full-length sequences of three heat stress response related genes: (1) the heat shock protein 70 (OvHSP70), (2) the heat shock transcription factor (OvHSF) , and (3) the heat shock factor-binding protein (OvHSBP) ofO.vulgaris. The OvHSP70, OvHSF, and OvHSBP proteins contained 2 222 bp, 2 264 bp, 841 bp that encoded for 635, 458 and 90 amino acids, respectively. The results of multiple sequence alignment showed that the amino acid sequences of OvHSP70 were highly conserved with respect to other species. Similarly, the DNA binding domain, the trimerization domain of OvHSF, and the coiled coil region of OvHSBP also had highly conserved regions. The real-time polymerase chain reaction (PCR) results indicated that OvHSP70 was temperature-dependent and time-dependent, showing a positive response to heat stress. On exposure to 28°C and to 30°C, the mRNA expression levels of OvHSF and OvHSBP were higher than those in the control group at 24°C. The mRNA expression of OvHSBP significantly increased with heat treatment at 26°C, while the mRNA expression of OvHSF decreased. The experimental results indicated that the expression of OvHSP70, OvHSF and OvHSBP were all sensitive to heat stress, which suggests that these three genes may play an important role forO.vulgaris in responding to environmental stress. Thus, this study sets a theoretical foundation for further in-depth studies on the molecular protective mechanisms of the heat response inO.vulgaris.

  10. Molecular karyotype and chromosomal localization of genes encoding ß-tubulin, cysteine proteinase, hsp 70 and actin in Trypanosoma rangeli

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    CB Toaldo

    2001-01-01

    Full Text Available The molecular karyotype of nine Trypanosoma rangeli strains was analyzed by contour-clamped homogeneous electric field electrophoresis, followed by the chromosomal localization of ß-tubulin, cysteine proteinase, 70 kDa heat shock protein (hsp 70 and actin genes. The T. rangeli strains were isolated from either insects or mammals from El Salvador, Honduras, Venezuela, Colombia, Panama and southern Brazil. Also, T. cruzi CL-Brener clone was included for comparison. Despite the great similarity observed among strains from Brazil, the molecular karyotype of all T. rangeli strains analyzed revealed extensive chromosome polymorphism. In addition, it was possible to distinguish T. rangeli from T. cruzi by the chromosomal DNA electrophoresis pattern. The localization of ß-tubulin genes revealed differences among T. rangeli strains and confirmed the similarity between the isolates from Brazil. Hybridization assays using probes directed to the cysteine proteinase, hsp 70 and actin genes discriminated T. rangeli from T. cruzi, proving that these genes are useful molecular markers for the differential diagnosis between these two species. Numerical analysis based on the molecular karyotype data revealed a high degree of polymorphism among T. rangeli strains isolated from southern Brazil and strains isolated from Central and the northern South America. The T. cruzi reference strain was not clustered with any T. rangeli strain.

  11. Molecular Mechanism of Allosteric Communication in Hsp70 Revealed by Molecular Dynamics Simulations

    OpenAIRE

    Chiappori, Federica; Merelli, Ivan; Colombo, Giorgio; Milanesi, Luciano; Morra, Giulia

    2012-01-01

    Author Summary Allostery, or the capability of proteins to respond to ligand binding events with a variation in structure or dynamics at a distant site, is a common feature for biomolecular function and regulation in a large number of proteins. Intra-protein connections and inter-residue coordinations underlie allosteric mechanisms and react to binding primarily through a finely tuned modulation of motions and structures at the microscopic scale. Hence, all-atom molecular dynamics simulations...

  12. Reduced Hsp70 and Glutamine in Pediatric Severe Malaria Anemia

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    Kempaiah, Prakasha; Dokladny, Karol; Karim, Zachary;

    2016-01-01

    decreased HSPA1A, a heat shock protein (Hsp) 70 coding gene. Hsp70 is a ubiquitous chaperone that regulates Nuclear Factor-kappa B (NF-κB) signaling and production of pro-inflammatory cytokines known to be important in malaria pathogenesis (e.g., IL-1β, IL-6 and TNF-α). Since the role of host Hsp70 in...... malaria pathogenesis is unexplored, we investigated Hsp70 and molecular pathways in children with SMA. Validation experiments revealed that leukocytic HSP70 transcripts were reduced in SMA relative to non-severe malaria, and that intraleukocytic hemozoin (PfHz) was associated with lower HSP70. HSP70 was...... correlated with reticulocyte production and Hb. Since glutamine (Gln) up-regulates Hsp70, modulates NF-κB activation, and attenuates over-expression of pro-inflammatory cytokines, circulating Gln was measured in children with malaria. Reduced Gln was associated with increased risk of developing SMA...

  13. The Assembly and Intermolecular Properties of the Hsp70-Tomm34-Hsp90 Molecular Chaperone Complex

    Czech Academy of Sciences Publication Activity Database

    Trčka, F.; Durech, M.; Hernychová, L.; Man, Petr; Müller, P.; Vojtěšek, B.

    2014-01-01

    Roč. 289, č. 14 (2014), s. 9887-9901. ISSN 0021-9258 R&D Projects: GA ČR(CZ) P301/11/1678; GA MZd(CZ) 00209805 Grant ostatní: Regional Center for Applied Molecular Oncology (CZ) CZ.1.05/2.1.00/03.0101 Institutional support: RVO:61388971 Keywords : HSP90 * chaperone * protein assembly Subject RIV: EE - Microbiology, Virology Impact factor: 4.573, year: 2014

  14. Sequence analysis of the 3’-untranslated region of HSP70 (type I genes in the genus Leishmania: its usefulness as a molecular marker for species identification

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    Requena Jose M

    2012-04-01

    Full Text Available Abstract Background The Leishmaniases are a group of clinically diverse diseases caused by parasites of the genus Leishmania. To distinguish between species is crucial for correct diagnosis and prognosis as well as for treatment decisions. Recently, sequencing of the HSP70 coding region has been applied in phylogenetic studies and for identifying of Leishmania species with excellent results. Methods In the present study, we analyzed the 3’-untranslated region (UTR of Leishmania HSP70-type I gene from 24 strains representing eleven Leishmania species in the belief that this non-coding region would have a better discriminatory capacity for species typing than coding regions. Results It was observed that there was a remarkable degree of sequence conservation in this region, even between species of the subgenus Leishmania and Viannia. In addition, the presence of many microsatellites was a common feature of the 3´-UTR of HSP70-I genes in the Leishmania genus. Finally, we constructed dendrograms based on global sequence alignments of the analyzed Leishmania species and strains, the results indicated that this particular region of HSP70 genes might be useful for species (or species complex typing, improving for particular species the discrimination capacity of phylogenetic trees based on HSP70 coding sequences. Given the large size variation of the analyzed region between the Leishmania and Viannia subgenera, direct visualization of the PCR amplification product would allow discrimination between subgenera, and a HaeIII-PCR-RFLP analysis might be used for differentiating some species within each subgenera. Conclusions Sequence and phylogenetic analyses indicated that this region, which is readily amplified using a single pair of primers from both Old and New World Leishmania species, might be useful as a molecular marker for species discrimination.

  15. Plasmodium falciparum Hsp70-z, an Hsp110 homologue, exhibits independent chaperone activity and interacts with Hsp70-1 in a nucleotide-dependent fashion.

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    Zininga, Tawanda; Achilonu, Ikechukwu; Hoppe, Heinrich; Prinsloo, Earl; Dirr, Heini W; Shonhai, Addmore

    2016-05-01

    The role of molecular chaperones, among them heat shock proteins (Hsps), in the development of malaria parasites has been well documented. Hsp70s are molecular chaperones that facilitate protein folding. Hsp70 proteins are composed of an N-terminal nucleotide binding domain (NBD), which confers them with ATPase activity and a C-terminal substrate binding domain (SBD). In the ADP-bound state, Hsp70 possesses high affinity for substrate and releases the folded substrate when it is bound to ATP. The two domains are connected by a conserved linker segment. Hsp110 proteins possess an extended lid segment, a feature that distinguishes them from canonical Hsp70s. Plasmodium falciparum Hsp70-z (PfHsp70-z) is a member of the Hsp110 family of Hsp70-like proteins. PfHsp70-z is essential for survival of malaria parasites and is thought to play an important role as a molecular chaperone and nucleotide exchange factor of its cytosolic canonical Hsp70 counterpart, PfHsp70-1. Unlike PfHsp70-1 whose functions are fairly well established, the structure-function features of PfHsp70-z remain to be fully elucidated. In the current study, we established that PfHsp70-z possesses independent chaperone activity. In fact, PfHsp70-z appears to be marginally more effective in suppressing protein aggregation than its cytosol-localized partner, PfHsp70-1. Furthermore, based on coimmunoaffinity chromatography and surface plasmon resonance analyses, PfHsp70-z associated with PfHsp70-1 in a nucleotide-dependent fashion. Our findings suggest that besides serving as a molecular chaperone, PfHsp70-z could facilitate the nucleotide exchange function of PfHsp70-1. These dual functions explain why it is essential for parasite survival. PMID:26894764

  16. The Complex Function of Hsp70 in Metastatic Cancer

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    Kata Juhasz

    2013-12-01

    Full Text Available Elevated expression of the inducible heat shock protein 70 (Hsp70 is known to correlate with poor prognosis in many cancers. Hsp70 confers survival advantage as well as resistance to chemotherapeutic agents, and promotes tumor cell invasion. At the same time, tumor-derived extracellular Hsp70 has been recognized as a “chaperokine”, activating antitumor immunity. In this review we discuss localization dependent functions of Hsp70 in the context of invasive cancer. Understanding the molecular principles of metastasis formation steps, as well as interactions of the tumor cells with the microenvironment and the immune system is essential for fighting metastatic cancer. Although Hsp70 has been implicated in different steps of the metastatic process, the exact mechanisms of its action remain to be explored. Known and potential functions of Hsp70 in controlling or modulating of invasion and metastasis are discussed.

  17. Molecular cloning, characterization and tissue profiles of a HSP70 cDNA in Siberian sturgeon Acipenser baerii%西伯利亚鲟热休克蛋白HSP70 cDNA的克隆、序列分析和组织分布

    Institute of Scientific and Technical Information of China (English)

    田照辉; 徐绍刚; 王巍; 胡红霞; 董颖; 宋超

    2012-01-01

    A full length cDNA encoding heat shock protein 70 (HSP70) was cloned from liver RNA in Siberian sturgeon (Acipenser baerii) ,and was found to contain 2 343 bp, including 5'untranslated region of 140 bp, 3'un-translated region of 256 bp and an open reading frame of 1 947 bp. The deduced 648 amino acids sequence with estimated molecular of 71 000 and theoretical isoelectric point of 5. 21 had high similarity with other vertebrate HSP70 genes in NCBI. Sequence analysis indicated that the HSP70 cDNA was consisted of three signature sequences IDL-GTTYS, IFDLGGGTFDVSrL and IVLVGGSTRIPKIQK belonging to HSP70 family. Phylogenetic tree revealed that Alligator mississippiensis, Xenopus laevis ,Ambystoma mexicanum and Sebrian Sturgeon showed a close genetic relationship. Using fluorescent real-time quantitative PCR.the HSP70 was expressed in the liver, gill, spleen, heart, intestine, muscle, gonad and brain of Siberian sturgeon, with the peak in the spleen and the minimum in the liver (P<0.05) , indicating that the HSP70 cDNA could be used in understanding of anti-stress mechanism in Siberian sturgeon and in genetic breeding improvement.%采用普通PCR和RACE技术克隆了西伯利亚鲟Acipenser baerii热休克蛋白HSP70 cDNA的全序列,该序列全长为2343 bp,其中5'非编码区为140 bp,3'非编码区为256 bp,可读编码框(ORF)为1947bp,编码为648个氨基酸.该氨基酸序列中含有HSP70家族的3个特征序列——IDLGTTYS、IFDLGGGTFDVSIL和ⅣLVGGSTRIPKIQK,细胞质特征性保守序列为EEVD,C端重复序列为GGMP.该cDNA序列与其它生物的HSP70 cDNA序列一样具有很高的相似性.系统发育树显示,西伯利亚鲟与非洲爪蛙蟾Xenopus laevis、密西西比短吻鳄Alligator mississippiensis、美西螈Ambystoma mexicanum的亲缘关系较近.实时定量分析结果表明,水温为17.5℃时,西伯利亚鲟肝脏、鳃、脾脏、心脏、肌肉、中肠、性腺、脑8种组织中均有HSP70表达,其中HSP70在脾脏中的表达

  18. Targeting Hsp70: A possible therapy for cancer.

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    Kumar, Sanjay; Stokes, James; Singh, Udai P; Scissum Gunn, Karyn; Acharya, Arbind; Manne, Upender; Mishra, Manoj

    2016-04-28

    In all organisms, heat-shock proteins (HSPs) provide an ancient defense system. These proteins act as molecular chaperones by assisting proper folding and refolding of misfolded proteins and aid in the elimination of old and damaged cells. HSPs include Hsp100, Hsp90, Hsp70, Hsp40, and small HSPs. Through its substrate-binding domains, Hsp70 interacts with wide spectrum of molecules, ranging from unfolded to natively folded and aggregated proteins, and provides cytoprotective role against various cellular stresses. Under pathophysiological conditions, the high expression of Hsp70 allows cells to survive with lethal injuries. Increased Hsp70, by interacting at several points on apoptotic signaling pathways, leads to inhibition of apoptosis. Elevated expression of Hsp70 in cancer cells may be responsible for tumorigenesis and for tumor progression by providing resistance to chemotherapy. In contrast, inhibition or knockdown of Hsp70 reduces the size of tumors and can cause their complete regression. Moreover, extracellular Hsp70 acts as an immunogen that participates in cross presentation of MHC-I molecules. The goals of this review are to examine the roles of Hsp70 in cancer and to present strategies targeting Hsp70 in the development of cancer therapeutics. PMID:26898980

  19. Effect of environmental impact to molecular expression of heat-shock protein (HSP70) in oyster Crassostrea gigas from Gamak bay, Korea.

    Science.gov (United States)

    Cho, Eun-Seob; Jeong, Hee-Dong

    2012-05-01

    The cDNA of Crassostrea gigas HSP70 was cloned and rapid amplification of cDNA (RACE) techniques were used. The full length of HSP70 cDNA was 2045 bp, consisting of a 5' terminal untranslated region (UTR) of 80 bp, a 3' terminal UTR 146 bp, and an open reading frame (ORF) of 1829 bp encoding deduced 620 amino acids. The HSP70 cDNA contained HSP70 family signatures, ATP-GTP binding site motif, tetrapeptide (GGMP) and conserved carboxyl terminal region (EEVD) at C-terminal of deduced amino acid sequence. BLAST analysis revealed that the HSP70 gene has an extreme similarity of 98.9% with C. gigas (AF144646). Northern blotting was used to examine the expression of HSP70 mRNA in the gill tissue of the oyster obtained from surface, middle and bottom layers. The HSP70 mRNA observed the samples taken from middle and bottom layers in September and February, but samples from the surface layer did not find a signal intensity of HSP70 mRNA transcript. Consequently, it seems that the oyster occurring middle and bottom layer have been stressed during the period of summer and winter, which is associated with the massive mortality in Gamak bay. PMID:23029911

  20. The genes coding for the hsp70(dnaK) molecular chaperone machine occur in the moderate thermophilic archaeon Methanosarcina thermophila TM-1

    DEFF Research Database (Denmark)

    Hofman-Bang, H Jacob Peider; Lange, Marianne; Ahring, Birgitte Kiær

    1999-01-01

    response by hsp70(dnaK), and a similar response by trkA. The data suggest that the moderate thermophile TM-1 has an active Hsp70(DnaK)-chaperone machine in contrast to hyperthermophilic archaea, and that trkA is a stress gene, inasmuch as it responds like classic heat-shock genes to stressors that induce a...

  1. Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself

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    Luong Michael

    2012-03-01

    Full Text Available Abstract Background Toll-like receptor 4 (TLR4 is activated by bacterial endotoxin, a prototypical pathogen-associated molecular pattern (PAMP. It has been suggested that TLR4 can also be activated by damage-associated molecular pattern (DAMP proteins such as HSP70. It remains a challenge to provide unequivocal evidence that DAMP proteins themselves play a role in TLR4 activation, as the DAMP proteins used are often contaminated with endotoxin and other TLR ligands introduced during protein expression and/or purification. Results Here we report that the activation of TLR4 on primary human macrophage cultures by recombinant HSP70 is not solely due to contaminating endotoxin. Polymyxin B pretreatment of HSP70 preparations to neutralize contaminating endotoxin caused significant reductions in the amount of TNF-α induced by the recombinant protein as determined by ELISA. However, digestion of HSP70 with Proteinase K-agarose beads also dramatically reduced the TNF-α response of macrophages to HSP70, while leaving levels of contaminating endotoxin largely unchanged relative to controls. Conclusions These results indicate that the stimulatory effect of recombinant HSP70 requires both the presence of endotoxin and structural integrity of the heat shock protein itself.

  2. Design of a fluorescence polarization assay platform for the study of human Hsp70

    OpenAIRE

    Kang, Yanlong; Taldone, Tony; Clement, Cristina C.; Fewell, Sheara W.; Aguirre, Julia; Brodsky, Jeffrey L.; Chiosis, Gabriela

    2008-01-01

    The 70 kDa heat shock proteins (Hsp70) are molecular chaperones that assist in folding of newly synthesized polypeptides, refolding or denaturation of misfolded proteins, and translocation of proteins across biological membranes. In addition, Hsp70 play regulatory roles in signal transduction, cell cycle, and apoptosis. Here we present a novel assay platform based on fluorescence polarization that is suitable for investigating the yet elusive molecular mechanics of human Hsp70 allosteric regu...

  3. The genes coding for the hsp70(dnaK) molecular chaperone machine occur in the moderate thermophilic archaeon Methanosarcina thermophila TM-1

    DEFF Research Database (Denmark)

    Hofman-Bang, H Jacob Peider; Lange, Marianne; Ahring, Birgitte Kiær

    1999-01-01

    response by hsp70(dnaK), and a similar response by trkA. The data suggest that the moderate thermophile TM-1 has an active Hsp70(DnaK)-chaperone machine in contrast to hyperthermophilic archaea, and that trkA is a stress gene, inasmuch as it responds like classic heat-shock genes to stressors that induce a...... typical heat-shock response. (C) 1999 Elsevier Science B.V. All rights reserved.......-negative bacteria - first described in the S-6 molecule and later found to be present in all homologs from archaea and Gram positives. The genes responded to a temperature elevation in a manner that demonstrated that they are heat-shock genes, functionally active in vivo. Ammonia also induced a heat-shock type of...

  4. High mobility group protein DSP1 negatively regulates HSP70 transcription in Crassostrea hongkongensis.

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    Miao, Zongyu; Xu, Delin; Cui, Miao; Zhang, Qizhong

    2016-06-10

    HSP70 acts mostly as a molecular chaperone and plays important roles in facilitating the folding of nascent peptides as well as the refolding or degradation of the denatured proteins. Under stressed conditions, the expression level of HSP70 is upregulated significantly and rapidly, as is known to be achieved by various regulatory factors controlling the transcriptional level. In this study, a high mobility group protein DSP1 was identified by DNA-affinity purification from the nuclear extracts of Crassostrea hongkongensis using the ChHSP70 promoter as a bait. The specific interaction between the prokaryotically expressed ChDSP1 and the FITC-labeled ChHSP70 promoter was confirmed by EMSA analysis. ChDSP1 was shown to negatively regulate ChHSP70 promoter expression by Luciferase Reporter Assay in the heterologous HEK293T cells. Both ChHSP70 and ChDSP1 transcriptions were induced by either thermal or CdCl2 stress, while the accumulated expression peaks of ChDSP1 were always slightly delayed when compared with that of ChHSP70. This indicates that ChDSP1 is involved, very likely to exert its suppressive role, in the recovery of the ChHSP70 expression from the induced level to its original state. This study is the first to report negative regulator of HSP70 gene transcription, and provides novel insights into the mechanisms controlling heat shock protein expression. PMID:27154224

  5. Roles of Hsp70s in Stress Responses of Microorganisms, Plants, and Animals

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    Anmin Yu

    2015-01-01

    Full Text Available Hsp70s (heat shock protein 70s are a class of molecular chaperones that are highly conserved and ubiquitous in organisms ranging from microorganisms to plants and humans. Most research on Hsp70s has focused on the mechanisms of their functions as molecular chaperones, but recently, studies on stress responses are coming to the forefront. Hsp70s play key roles in cellular development and protecting living organisms from environmental stresses such as heat, drought, salinity, acidity, and cold. Moreover, functions of human Hsp70s are related to diseases including neurological disorders, cancer, and virus infection. In this review, we provide an overview of the specific roles of Hsp70s in response to stress, particularly abiotic stress, in all living organisms.

  6. Diversity of cytosolic HSP70 Heat Shock Protein from decapods and their phylogenetic placement within Arthropoda.

    Science.gov (United States)

    Baringou, Stephane; Rouault, Jacques-Deric; Koken, Marcel; Hardivillier, Yann; Hurtado, Luis; Leignel, Vincent

    2016-10-10

    The 70kDa heat shock proteins (HSP70) are considered the most conserved members of the HSP family. These proteins are primordial to the cell, because of their implications in many cellular pathways (e. g., development, immunity) and also because they minimize the effects of multiple stresses (e. g., temperature, pollutants, salinity, radiations). In the cytosol, two ubiquitous HSP70s with either a constitutive (HSC70) or an inducible (HSP70) expression pattern are found in all metazoan species, encoded by 5 or 6 genes (Drosophila melanogaster or yeast and human respectively). The cytosolic HSP70 protein family is considered a major actor in environmental adaptation, and widely used in ecology as an important biomarker of environmental stress. Nevertheless, the diversity of cytosolic HSP70 remains unclear amongst the Athropoda phylum, especially within decapods. Using 122 new and 311 available sequences, we carried out analyses of the overall cytosolic HSP70 diversity in arthropods (with a focus on decapods) and inferred molecular phylogenies. Overall structural and phylogenetic analyses showed a surprisingly high diversity in cytosolic HSP70 and revealed the existence of several unrecognised groups. All crustacean HSP70 sequences present signature motifs and molecular weights characteristic of non-organellar HSP70, with multiple specific substitutions in the protein sequence. The cytosolic HSP70 family in arthropods appears to be constituted of at least three distinct groups (annotated as A, B and C), which comprise several subdivisions, including both constitutive and inducible forms. Group A is constituted by several classes of Arthropods, while group B and C seem to be specific to Malacostraca and Hexapoda/Chelicerata, respectively. The HSP70 organization appeared much more complex than previously suggested, and far beyond a simple differentiation according to their expression pattern (HSC70 versus HSP70). This study proposes a new classification of cytosolic

  7. Regulation of Hsp70 function by nucleotide-exchange factors

    OpenAIRE

    Gowda, Naveen Kumar Chandappa

    2016-01-01

    Protein folding is the process in which polypeptides in their non-native states attain the unique folds of their native states. Adverse environmental conditions and genetic predisposition challenge the folding process and accelerate the production of proteotoxic misfolded proteins. Misfolded proteins are selectively recognized and removed from the cell by processes of protein quality control (PQC). In PQC molecular chaperones of the Heat shock protein 70 kDa (Hsp70) family play important role...

  8. A novel Hsp70 inhibitor prevents cell intoxication with the actin ADP-ribosylating Clostridium perfringens iota toxin.

    Science.gov (United States)

    Ernst, Katharina; Liebscher, Markus; Mathea, Sebastian; Granzhan, Anton; Schmid, Johannes; Popoff, Michel R; Ihmels, Heiko; Barth, Holger; Schiene-Fischer, Cordelia

    2016-01-01

    Hsp70 family proteins are folding helper proteins involved in a wide variety of cellular pathways. Members of this family interact with key factors in signal transduction, transcription, cell-cycle control, and stress response. Here, we developed the first Hsp70 low molecular weight inhibitor specifically targeting the peptide binding site of human Hsp70. After demonstrating that the inhibitor modulates the Hsp70 function in the cell, we used the inhibitor to show for the first time that the stress-inducible chaperone Hsp70 functions as molecular component for entry of a bacterial protein toxin into mammalian cells. Pharmacological inhibition of Hsp70 protected cells from intoxication with the binary actin ADP-ribosylating iota toxin from Clostridium perfringens, the prototype of a family of enterotoxins from pathogenic Clostridia and inhibited translocation of its enzyme component across cell membranes into the cytosol. This finding offers a starting point for novel therapeutic strategies against certain bacterial toxins. PMID:26839186

  9. Detection of irradiation-induced, membrane heat shock protein 70 (Hsp70) in mouse tumors using Hsp70 Fab fragment

    International Nuclear Information System (INIS)

    Background and purpose: The major stress-inducible heat shock protein 70 (Hsp70) is frequently overexpressed in highly aggressive tumors, and elevated intracellular Hsp70 levels mediate protection against apoptosis. Following therapeutic intervention, such as ionizing irradiation, translocation of cytosolic Hsp70 to the plasma membrane is selectively increased in tumor cells and therefore, membrane Hsp70 might serve as a therapy-inducible, tumor-specific target structure. Materials and methods: Based on the IgG1 mouse monoclonal antibody (mAb) cmHsp70.1, we produced the Hsp70-specific recombinant Fab fragment (Hsp70 Fab), as an imaging tool for the detection of membrane Hsp70 positive tumor cells in vitro and in vivo. Results: The binding characteristics of Hsp70 Fab towards mouse colon (CT26) and pancreatic (1048) carcinoma cells at 4 deg. C were comparable to that of cmHsp70.1 mAb, as determined by flow cytometry. Following a temperature shift to 37 deg. C, Hsp70 Fab rapidly translocates into subcellular vesicles of mouse tumor cells. Furthermore, in tumor-bearing mice Cy5.5-conjugated Hsp70 Fab, but not unrelated IN-1 control Fab fragment (IN-1 ctrl Fab), gradually accumulates in CT26 tumors between 12 and 55 h after i.v. injection. Conclusions: In summary, the Hsp70 Fab provides an innovative, low immunogenic tool for imaging of membrane Hsp70 positive tumors, in vivo.

  10. PROFILES OF GENE EXPRESSION ASSOCIATED WITH TETRACYCLINE OVER EXPRESSION OF HSP70 IN MCF-7 BREAST CANCER CELLS

    Science.gov (United States)

    Profiles of gene expression associated with tetracycline over expression of HSP70 in MCF-7 breast cancer cells. Heat shock proteins (HSPs) protect cells from damage through their function as molecular chaperones. Some cancers reveal high levels of HSP70 expression in asso...

  11. Hsp70 chaperones accelerate protein translocation and the unfolding of stable protein aggregates by entropic pulling.

    Science.gov (United States)

    De Los Rios, Paolo; Ben-Zvi, Anat; Slutsky, Olga; Azem, Abdussalam; Goloubinoff, Pierre

    2006-04-18

    Hsp70s are highly conserved ATPase molecular chaperones mediating the correct folding of de novo synthesized proteins, the translocation of proteins across membranes, the disassembly of some native protein oligomers, and the active unfolding and disassembly of stress-induced protein aggregates. Here, we bring thermodynamic arguments and biochemical evidences for a unifying mechanism named entropic pulling, based on entropy loss due to excluded-volume effects, by which Hsp70 molecules can convert the energy of ATP hydrolysis into a force capable of accelerating the local unfolding of various protein substrates and, thus, perform disparate cellular functions. By means of entropic pulling, individual Hsp70 molecules can accelerate unfolding and pulling of translocating polypeptides into mitochondria in the absence of a molecular fulcrum, thus settling former contradictions between the power-stroke and the Brownian ratchet models for Hsp70-mediated protein translocation across membranes. Moreover, in a very different context devoid of membrane and components of the import pore, the same physical principles apply to the forceful unfolding, solubilization, and assisted native refolding of stable protein aggregates by individual Hsp70 molecules, thus providing a mechanism for Hsp70-mediated protein disaggregation. PMID:16606842

  12. Molecular characterization of three heat shock protein 70 genes and their expression profiles under thermal stress in the citrus red mite.

    Science.gov (United States)

    Yang, Li-Hong; Jiang, Hong-Bo; Liu, Yong-Hua; Dou, Wei; Wang, Jin-Jun

    2012-04-01

    Three heat shock protein 70 family transcripts, named PcHsp70-1, PcHsp70-2 and PcHsp70-3, were isolated from the citrus red mite, Panonychus citri. PcHsp70-1, PcHsp70-2, and PcHsp70-3 contained an open reading frame of 1977, 1968, and 2028 nucleotides that encoded 658, 655 and 675 amino acid residues, respectively. Comparison of deduced amino acid sequences of PcHsp70-1 and PcHsp70-2 showed 86.34% identity, while the amino acid sequence of PcHsp70-3 was only 57.39 and 58.75% identical to that of PcHsp70-1 and PcHsp70-2, respectively. Sequences and phylogenetic analyses suggested that PcHsp70-1 and PcHsp70-2 were cytosolic Hsps, whereas PcHsp70-3 was located in ER (endoplasmic reticulum). To accurately validate mRNA expression profiles of the three Hsp70s under thermal stress conditions, seven housekeeping genes were evaluated. Alpha-tubulin and RpII were selected as optimal endogenous references for cold shock and heat shock conditions, respectively. Real-time quantitative RT-PCR revealed that only the mRNA expression of PcHsp70-2 was up-regulated under heat shocks, and all of the three Hsp70s were constitutively expressed under cold shocks. The results suggest that the three Hsp70s were more critical to coping with heat than cold shocks. PMID:21717055

  13. The Malarial Exported PFA0660w Is an Hsp40 Co-Chaperone of PfHsp70-x.

    Directory of Open Access Journals (Sweden)

    Michael O Daniyan

    Full Text Available Plasmodium falciparum, the human pathogen responsible for the most dangerous malaria infection, survives and develops in mature erythrocytes through the export of proteins needed for remodelling of the host cell. Molecular chaperones of the heat shock protein (Hsp family are prominent members of the exportome, including a number of Hsp40s and a Hsp70. PFA0660w, a type II Hsp40, has been shown to be exported and possibly form a complex with PfHsp70-x in the infected erythrocyte cytosol. However, the chaperone properties of PFA0660w and its interaction with human and parasite Hsp70s are yet to be investigated. Recombinant PFA0660w was found to exist as a monomer in solution, and was able to significantly stimulate the ATPase activity of PfHsp70-x but not that of a second plasmodial Hsp70 (PfHsp70-1 or a human Hsp70 (HSPA1A, indicating a potential specific functional partnership with PfHsp70-x. Protein binding studies in the presence and absence of ATP suggested that the interaction of PFA0660w with PfHsp70-x most likely represented a co-chaperone/chaperone interaction. Also, PFA0660w alone produced a concentration-dependent suppression of rhodanese aggregation, demonstrating its chaperone properties. Overall, we have provided the first biochemical evidence for the possible role of PFA0660w as a chaperone and as co-chaperone of PfHsp70-x. We propose that these chaperones boost the chaperone power of the infected erythrocyte, enabling successful protein trafficking and folding, and thereby making a fundamental contribution to the pathology of malaria.

  14. Species- and Strain-Specific Adaptation of the HSP70 Super Family in Pathogenic Trypanosomatids.

    Science.gov (United States)

    Drini, Sima; Criscuolo, Alexis; Lechat, Pierre; Imamura, Hideo; Skalický, Tomáš; Rachidi, Najma; Lukeš, Julius; Dujardin, Jean-Claude; Späth, Gerald F

    2016-01-01

    All eukaryotic genomes encode multiple members of the heat shock protein 70 (HSP70) family, which evolved distinctive structural and functional features in response to specific environmental constraints. Phylogenetic analysis of this protein family thus can inform on genetic and molecular mechanisms that drive species-specific environmental adaptation. Here we use the eukaryotic pathogen Leishmania spp. as a model system to investigate the evolution of the HSP70 protein family in an early-branching eukaryote that is prone to gene amplification and adapts to cytotoxic host environments by stress-induced and chaperone-dependent stage differentiation. Combining phylogenetic and comparative analyses of trypanosomatid genomes, draft genome of Paratrypanosoma and recently published genome sequences of 204 L. donovani field isolates, we gained unique insight into the evolutionary dynamics of the Leishmania HSP70 protein family. We provide evidence for (i) significant evolutionary expansion of this protein family in Leishmania through gene amplification and functional specialization of highly conserved canonical HSP70 members, (ii) evolution of trypanosomatid-specific, non-canonical family members that likely gained ATPase-independent functions, and (iii) loss of one atypical HSP70 member in the Trypanosoma genus. Finally, we reveal considerable copy number variation of canonical cytoplasmic HSP70 in highly related L. donovani field isolates, thus identifying this locus as a potential hot spot of environment-genotype interaction. Our data draw a complex picture of the genetic history of HSP70 in trypanosomatids that is driven by the remarkable plasticity of the Leishmania genome to undergo massive intra-chromosomal gene amplification to compensate for the absence of regulated transcriptional control in these parasites. PMID:27371955

  15. Targeting membrane heat-shock protein 70 (Hsp70) on tumors by cmHsp70.1 antibody

    Science.gov (United States)

    Stangl, Stefan; Gehrmann, Mathias; Riegger, Julia; Kuhs, Kristin; Riederer, Isabelle; Sievert, Wolfgang; Hube, Kathrin; Mocikat, Ralph; Dressel, Ralf; Kremmer, Elisabeth; Pockley, Alan G.; Friedrich, Lars; Vigh, Laszlo; Skerra, Arne; Multhoff, Gabriele

    2011-01-01

    Immunization of mice with a 14-mer peptide TKDNNLLGRFELSG, termed “TKD,” comprising amino acids 450–461 (aa450–461) in the C terminus of inducible Hsp70, resulted in the generation of an IgG1 mouse mAb cmHsp70.1. The epitope recognized by cmHsp70.1 mAb, which has been confirmed to be located in the TKD sequence by SPOT analysis, is frequently detectable on the cell surface of human and mouse tumors, but not on isogenic cells and normal tissues, and membrane Hsp70 might thus serve as a tumor-specific target structure. As shown for human tumors, Hsp70 is associated with cholesterol-rich microdomains in the plasma membrane of mouse tumors. Herein, we show that the cmHsp70.1 mAb can selectively induce antibody-dependent cellular cytotoxicity (ADCC) of membrane Hsp70+ mouse tumor cells by unstimulated mouse spleen cells. Tumor killing could be further enhanced by activating the effector cells with TKD and IL-2. Three consecutive injections of the cmHsp70.1 mAb into mice bearing CT26 tumors significantly inhibited tumor growth and enhanced the overall survival. These effects were associated with infiltrations of NK cells, macrophages, and granulocytes. The Hsp70 specificity of the ADCC response was confirmed by preventing the antitumor response in tumor-bearing mice by coinjecting the cognate TKD peptide with the cmHsp70.1 mAb, and by blocking the binding of cmHsp70.1 mAb to CT26 tumor cells using either TKD peptide or the C-terminal substrate-binding domain of Hsp70. PMID:21187371

  16. Targeting membrane heat-shock protein 70 (Hsp70) on tumors by cmHsp70.1 antibody

    OpenAIRE

    Stangl, Stefan; Gehrmann, Mathias; Riegger, Julia; Kuhs, Kristin; Riederer, Isabelle; Sievert, Wolfgang; Hube, Kathrin; Mocikat, Ralph; Dressel, Ralf; Kremmer, Elisabeth; Pockley, Alan G.; Friedrich, Lars; Vigh, Laszlo; Skerra, Arne; Multhoff, Gabriele

    2010-01-01

    Immunization of mice with a 14-mer peptide TKDNNLLGRFELSG, termed “TKD,” comprising amino acids 450–461 (aa450–461) in the C terminus of inducible Hsp70, resulted in the generation of an IgG1 mouse mAb cmHsp70.1. The epitope recognized by cmHsp70.1 mAb, which has been confirmed to be located in the TKD sequence by SPOT analysis, is frequently detectable on the cell surface of human and mouse tumors, but not on isogenic cells and normal tissues, and membrane Hsp70 might thus serve as a tumor-s...

  17. SSI1 encodes a novel Hsp70 of the Saccharomyces cerevisiae endoplasmic reticulum.

    OpenAIRE

    Baxter, B K; James, P; Evans, T.(Department of Physics, University of Oxford, Oxford, UK); Craig, E A

    1996-01-01

    The endoplasmic reticulum (ER) of the budding yeast Saccharomyces cerevisiae contains a well-characterized, essential member of the Hsp70 family of molecular chaperones, Kar2p. Kar2p has been shown to be involved in the translocation of proteins into the ER as well as the proper folding of proteins in that compartment. We report the characterization of a novel Hsp70 of the ER, Ssi1p. Ssi1p, which shares 24% of the amino acids of Kar2p, is not essential for growth under normal conditions. Howe...

  18. Imaging of Hsp70-positive tumors with cmHsp70.1 antibody-conjugated gold nanoparticles

    Directory of Open Access Journals (Sweden)

    Gehrmann MK

    2015-09-01

    Full Text Available Mathias K Gehrmann,1 Melanie A Kimm,2 Stefan Stangl,1 Thomas E Schmid,1 Peter B Noël,2 Ernst J Rummeny,2 Gabriele Multhoff11Department of Radiation Oncology, 2Department of Diagnostic and Interventional Radiology, Klinikum rechts der Isar, Technische Universität München, Munich, GermanyAbstract: Real-time imaging of small tumors is still one of the challenges in cancer diagnosis, prognosis, and monitoring of clinical outcome. Targeting novel biomarkers that are selectively expressed on a large variety of different tumors but not normal cells has the potential to improve the imaging capacity of existing methods such as computed tomography. Herein, we present a novel technique using cmHsp70.1 monoclonal antibody-conjugated spherical gold nanoparticles for quantification of the targeted uptake of gold nanoparticles into membrane Hsp70-positive tumor cells. Upon binding, cmHsp70.1-conjugated gold nanoparticles but not nanoparticles coupled to an isotype-matched IgG1 antibody or empty nanoparticles are rapidly taken up by highly malignant Hsp70 membrane-positive mouse tumor cells. After 24 hours, the cmHsp70.1-conjugated gold nanoparticles are found to be enriched in the perinuclear region. Specificity for membrane Hsp70 was shown by using an Hsp70 knockout tumor cell system. Toxic side effects of the cmHsp70.1-conjugated nanoparticles are not observed at a concentration of 1–10 µg/mL. Experiments are ongoing to evaluate whether cmHsp70.1 antibody-conjugated gold nanoparticles are suitable for the detection of membrane-Hsp70-positive tumors in vivo.Keywords: heat shock protein 70, tumor biomarker, theranostics, multimodal CT, multispectral CT, k-edge

  19. Inducible HSP70 Protects Radiation-Induced Salivary Gland Damage

    International Nuclear Information System (INIS)

    Irradiation (IR) delivered to the head and neck is a common treatment for malignancies. Salivary glands in the irradiation field are severely damaged, and consequently this resulted in marked salivary hypofunction. While the exact mechanism of salivary gland damage remains enigmatic, fluid secreting acinar cells are lost, and saliva output is dramatically reduced. Previously we have reported that inducible heat shock protein 70 (HSP70i) induced radioresistance in vitro. Moreover, HSP70i localized to salivary glands by gene transfer has great potential for the treatment of salivary gland. Herein, we investigated whether HSP70 can use as radio protective molecules for radiation-induced salivary gland damage in vivo

  20. Pathways of allosteric regulation in Hsp70 chaperones.

    Science.gov (United States)

    Kityk, Roman; Vogel, Markus; Schlecht, Rainer; Bukau, Bernd; Mayer, Matthias P

    2015-01-01

    Central to the protein folding activity of Hsp70 chaperones is their ability to interact with protein substrates in an ATP-controlled manner, which relies on allosteric regulation between their nucleotide-binding (NBD) and substrate-binding domains (SBD). Here we dissect this mechanism by analysing mutant variants of the Escherichia coli Hsp70 DnaK blocked at distinct steps of allosteric communication. We show that the SBD inhibits ATPase activity by interacting with the NBD through a highly conserved hydrogen bond network, and define the signal transduction pathway that allows bound substrates to trigger ATP hydrolysis. We identify variants deficient in only one direction of allosteric control and demonstrate that ATP-induced substrate release is more important for chaperone activity than substrate-stimulated ATP hydrolysis. These findings provide evidence of an unexpected dichotomic allostery mechanism in Hsp70 chaperones and provide the basis for a comprehensive mechanical model of allostery in Hsp70s. PMID:26383706

  1. Pathways of allosteric regulation in Hsp70 chaperones

    OpenAIRE

    Kityk, Roman; Vogel, Markus; Schlecht, Rainer; Bukau, Bernd; Mayer, Matthias P

    2015-01-01

    Central to the protein folding activity of Hsp70 chaperones is their ability to interact with protein substrates in an ATP-controlled manner, which relies on allosteric regulation between their nucleotide-binding (NBD) and substrate-binding domains (SBD). Here we dissect this mechanism by analysing mutant variants of the Escherichia coli Hsp70 DnaK blocked at distinct steps of allosteric communication. We show that the SBD inhibits ATPase activity by interacting with the NBD through a highly ...

  2. Molecular cloning of hsf1 and hsbp1 cDNAs, and the expression of hsf1, hsbp1 and hsp70 under heat stress in the sea cucumber Apostichopus japonicus.

    Science.gov (United States)

    Xu, Dongxue; Sun, Lina; Liu, Shilin; Zhang, Libin; Yang, Hongsheng

    2016-08-01

    The heat shock response (HSR) is known for the elevated synthesis of heat shock proteins (HSPs) under heat stress, which is mediated primarily by heat shock factor 1 (HSF1). Heat shock factor binding protein 1 (HSBP1) and feedback control of heat shock protein 70 (HSP70) are major regulators of the activity of HSF1. We obtained full-length cDNA of genes hsf1 and hsbp1 in the sea cucumber Apostichopus japonicus, which are the second available for echinoderm (after Strongylocentrotus purpuratus), and the first available for holothurian. The full-length cDNA of hsf1 was 2208bp, containing a 1326bp open reading frame encoding 441 amino acids. The full-length cDNA of hsbp1 was 2850bp, containing a 225bp open reading frame encoding 74 amino acids. The similarities of A. japonicus HSF1 with other species are low, and much higher similarity identities of A. japonicus HSBP1 were shared. Phylogenetic trees showed that A. japonicus HSF1 and HSBP1 were clustered with sequences from S. purpuratus, and fell into distinct clades with sequences from mollusca, arthropoda and vertebrata. Analysis by real-time PCR showed hsf1 and hsbp1 mRNA was expressed constitutively in all tissues examined. The expression of hsf1, hsbp1 and hsp70 in the intestine at 26°C was time-dependent. The results of this study might provide new insights into the regulation of heat shock response in this species. PMID:26952354

  3. Increased hepatotoxicity of acetaminophen in Hsp70i knockout mice

    International Nuclear Information System (INIS)

    The effect of the inducible forms of 70 kDa heat shock protein (Hsp70i) on acetaminophen (APAP) hepatotoxicity was assessed in an Hsp70i knockout mouse model. Absence of the Hsp70i protein in liver was verified by monitoring Hsp levels in knockout and control mice after heat stress (41.5 oC water bath immersion for 30 min). Hsp70i knockout mice were more susceptible to APAP-induced hepatotoxicity than controls, as indicated by elevated serum alanine aminotransferase activities 24 and 48 h after the APAP dose. Increased APAP hepatotoxicity in knockout mice was verified by morphological evaluation of liver sections. The difference in toxic response to APAP between knockout and control strain mice could not be attributed to differences in APAP bioactivation, assessed by measurement of CYP2E1 and glutathione S-transferase activities, hepatic nonprotein sulfhydryl content, or covalent binding of reactive APAP metabolites to proteins. Pretreatment with transient hyperthermia to produce a general upregulation of Hsps resulted in decreased APAP hepatotoxicity in both the knockout and control strains. Among thermally-pretreated mice, hepatotoxicity of APAP was greater in the knockouts compared with the control strain. These observations suggest that increased Hsp70i expression in response to APAP acts to limit the extent of tissue injury. Results further suggest that other factors related to heat stress can also contribute to protection against APAP toxicity

  4. Expression of HSP70 in cerebral ischemia and neuroprotetive action of hypothermia and ketoprofen Expressão de HSP70 na isquemia cerebral e a ação neuroprotetora da hipotermia e do cetoprofeno

    Directory of Open Access Journals (Sweden)

    Daniela Pretti da Cunha Tirapelli

    2010-08-01

    Full Text Available Heat shock proteins (HSPs are molecular chaperones that bind to other proteins to shepherd them across membranes and direct them to specific locations within a cell. Several injurious stimuli can induce Hsp70 expression, including ischemia. This study aimed to investigate the pattern of expression of protein (immunohistochemistry and gene (real-time PCR Hsp70 in experimental focal cerebral ischemia in rats by occlusion of the middle cerebral artery for 1 hour and the role of neuroprotection with hypothermia (H and ketoprofen (K. The infarct volume was measured using morphometric analysis defined by triphenyl tetrazolium chloride. It was observed increases in the protein (p=0.0001 and gene (p=0.0001 Hsp70 receptor in the ischemic areas that were reduced by H (protein and gene: pProteínas de choque térmico (HSPs são chaperones moleculares que se ligam a outras proteínas para atravessar as membranas e encaminhá-las para locais específicos dentro de uma célula. Vários estímulos nocivos podem induzir a expressão de Hsp70, incluindo isquemia. Este estudo teve como objetivo investigar o padrão de expressão protéica (imunohistoquímica e gênica (PCR em tempo real de Hsp70 na isquemia cerebral focal experimental em ratos pela oclusão da artéria cerebral média durante 1 hora e o papel da neuroproteção com hipotermia (H e cetoprofeno (C. O volume de infarto foi calculado através da análise morfométrica definido por cloreto de trifenil tetrazólio. Foi observado aumento na expressão proteína (p=0,0001 e gênica (p=0,0001 de Hsp70 nas áreas isquêmicas que foram reduzidas pela H (proteína e gene: p<0,05, C (proteína: p<0,001 e H+K (proteína: p<0,01 e gene: p<0,05. O aumento de Hsp70 na área isquêmica sugere que a neuroexcitotoxicidade mediada pela Hsp70 desempenha um papel importante na morte celular e que o efeito neuroprotetor tanto da H quanto do C está diretamente envolvido com a Hsp70.

  5. Hsp70 enhances presentation of FMDV antigen to bovine CD4+ T cells in vitro

    OpenAIRE

    McLaughlin, Kerry; Seago, Julian; Robinson, Lucy; Kelly, Charles; Charleston, Bryan

    2010-01-01

    International audience Foot-and-mouth disease virus (FMDV) is the causative agent of a highly contagious acute vesicular disease affecting cloven-hoofed animals, including cattle, sheep and pigs. The current vaccine induces a rapid humoral response, but the duration of the protective antibody response is variable, possibly associated with a variable specific CD4+ T cell response. We investigated the use of heat shock protein 70 (Hsp70) as a molecular chaperone to target viral antigen to th...

  6. Involvement of p27CIP/KIP in HSP25 or inducible HSP70 mediated adaptive response by low dose radiation

    International Nuclear Information System (INIS)

    ThermoResistant (TR) clones of Radiation-Induced Fibrosarcoma (RIF) cells have been reported to show an adaptive response to 1cGy of low dose radiation, and HSP25 and inducible HSP70 are involved in this process. In this study, to further elucidate the mechanism by which HSP25 and inducible HSP70 regulate the adaptive response, HSP25 or inducible HSP70 overexpressed RIF cells were irradiated with 1cGy and the cell cycle was analyzed. HSP25 or inducible HSP70 overexpressed cells together with TR cells showed increased G1 phase after 1cGy irradiation, while RIF cells did not. [3H]-Thymidine and BrdU incorporation also indicated that both HSP25 and inducible HSP70 are involved in G1 arrest after 1cGy irradiation. Molecular analysis revealed upregulation of p27Cip/Kip protein in HSP25 and inducible HSP70 overexpressed cells, and cotransfection of p27Cip/Kip antisense abolished the induction of the adaptive response and 1cGy-mediated G1 arrest. The above results indicate that induction of an adaptive response by HSP25 and inducible HSP70 is mediated by upregulation of p27Cip/Kip protein, resulting in low dose radiation-induced G1 arrest

  7. p27Cip/Kip is involved in hsp25 or inducible hsp70 mediated adaptive response by low dose radiation

    International Nuclear Information System (INIS)

    Thermoresistant (TR) clone of radiation-induced fibrosarcoma (RIF) cells have been reported to show adaptive response to 1cGy of low dose radiation, and hsp25 and inducible hsp70 are involved in this process. In the present study, to further elucidate the mechanism of how hsp25 and inducible hsp70 regulate the adaptive response, hsp25 or inducible hsp70 overexpressed RIF cells were irradiated with 1cGy and cell cycle was analyzed. Hsp25 or inducible hsp70 overexpressed cells as well as TR cells showed increase of G1 phase population after γ-irradiation at 1cGy, while the parent RIF cells did not. [3H]-Thymidine and BrdU incorporation also indicated that both hsp25 and inducible hsp70 were involved in G1 arrest after 1cGy irradiation. Molecular analysis revealed upregulation of p27Cip/Kip protein in hsp25 and inducible hsp70 overexpressed cells, and cotransfection of p27Cip/Kip antisense abolished the induction of adaptive response and 1cGy-mediated G1 arrest. The above results indicate that induction of adaptive response by hsp25 and inducible hsp70 is mediated by upregulation of p27Cip/Kip protein, resulting in low dose radiation-induced G1 arrest. (author)

  8. HSP70 induction during baculovirus infection

    Science.gov (United States)

    Baculoviruses are arthropod-specific double-stranded DNA viruses that have been employed as bio-insecticides against crop pests and to produce heterologous proteins in baculovirus expression systems. Although a consensus has emerged on the dominant molecular events driving baculovirus replication i...

  9. Possible role of Hsp70 in autoantigens shuttling to dermo-epidermal junction in systemic lupus erythematosus

    Directory of Open Access Journals (Sweden)

    E. Avalos-Díaz

    2011-09-01

    Full Text Available Under stress cells and tissues perform a series of physiologic adjustments, including the heat shock protein production (Hsp, these proteins play a role as molecular chaperons, they are divided in five main families: Hsp27, Hsp 60, Hsp70, Hsp90 and Hsp110 (1. Hsp are constitutively induced or expressed to support the correct folding of novel proteins or to refold damaged proteins (2. Hsp70 is the most representative member of Hsp family. It is largely distributed in the majority of eukaryotic cell compartments and interact with the cytoskeleton during translocation of proteins across membranes (3.

  10. Characterization and SNP variation analysis of a HSP70 gene from miiuy croaker and its expression as related to bacterial challenge and heat shock.

    Science.gov (United States)

    Wei, Tao; Sun, Yuena; Shi, Ge; Wang, Rixin; Xu, Tianjun

    2012-09-01

    Heat shock proteins (HSPs) play crucial roles in the immune response of vertebrates. In order to study immune defense mechanism of heat shock protein gene in miiuy croaker (Miichthys miiuy), a cDNA encoding heat shock protein 70 (designated Mimi-HSP70) gene was cloned from miiuy croaker. The cDNA was 2195 bp in length, consisting of an open reading frame (ORF) of 1917 bp encoding a polypeptide of 638 amino acids with estimated molecular mass of 70.3 kDa and theoretical isoelectric point of 5.55. Genomic DNA structure analysis revealed that the Mimi-HSP70 gene contain no introns in coding region and four SNPs with 373 C/T, 789 G/A, 1005 C/T, and 1185 G/A were detected by direct sequencing of 20 samples from six different populations. BLAST analysis, structure comparison and phylogenetic analysis indicated that Mimi-HSP70 should be an inducible cytosolic member of the HSP70 family. The deduced amino acid sequence of Mimi-HSP70 had 82.4%-92.2% identity with those of vertebrate. A real-time quantitative RT-PCR demonstrated that the HSP70 gene was ubiquitously expressed in ten normal tissues. Under different temperature shock stress, the expression of Mimi-HSP70 gene in miiuy croaker increased at first and then decreased with the rise of temperature, finally, reached a maximum level in liver, spleen and kidney tissues. Infection of miiuy croaker with Vibrio anguillarum resulted in significant changes expression of Mimi-HSP70 gene in the immune-related tissues. These results indicated that expression analysis of Mimi-HSP70 gene provide theoretical basis to further study the mechanism of anti-adverseness in the miiuy croaker. PMID:22750024

  11. In vivo generation of neurotoxic prion protein: role for hsp70 in accumulation of misfolded isoforms.

    Directory of Open Access Journals (Sweden)

    Pedro Fernandez-Funez

    2009-06-01

    Full Text Available Prion diseases are incurable neurodegenerative disorders in which the normal cellular prion protein (PrP(C converts into a misfolded isoform (PrP(Sc with unique biochemical and structural properties that correlate with disease. In humans, prion disorders, such as Creutzfeldt-Jakob disease, present typically with a sporadic origin, where unknown mechanisms lead to the spontaneous misfolding and deposition of wild type PrP. To shed light on how wild-type PrP undergoes conformational changes and which are the cellular components involved in this process, we analyzed the dynamics of wild-type PrP from hamster in transgenic flies. In young flies, PrP demonstrates properties of the benign PrP(C; in older flies, PrP misfolds, acquires biochemical and structural properties of PrP(Sc, and induces spongiform degeneration of brain neurons. Aged flies accumulate insoluble PrP that resists high concentrations of denaturing agents and contains PrP(Sc-specific conformational epitopes. In contrast to PrP(Sc from mammals, PrP is proteinase-sensitive in flies. Thus, wild-type PrP rapidly converts in vivo into a neurotoxic, protease-sensitive isoform distinct from prototypical PrP(Sc. Next, we investigated the role of molecular chaperones in PrP misfolding in vivo. Remarkably, Hsp70 prevents the accumulation of PrP(Sc-like conformers and protects against PrP-dependent neurodegeneration. This protective activity involves the direct interaction between Hsp70 and PrP, which may occur in active membrane microdomains such as lipid rafts, where we detected Hsp70. These results highlight the ability of wild-type PrP to spontaneously convert in vivo into a protease-sensitive isoform that is neurotoxic, supporting the idea that protease-resistant PrP(Sc is not required for pathology. Moreover, we identify a new role for Hsp70 in the accumulation of misfolded PrP. Overall, we provide new insight into the mechanisms of spontaneous accumulation of neurotoxic PrP and uncover

  12. Hsp70 stabilizes lysosomes and reverts Niemann-Pick disease-associated lysosomal pathology

    DEFF Research Database (Denmark)

    Kirkegaard, Thomas; Roth, Anke G; Petersen, Nikolaj H T;

    2010-01-01

    . In acidic environments Hsp70 binds with high affinity and specificity to BMP, thereby facilitating the BMP binding and activity of acid sphingomyelinase (ASM). The inhibition of the Hsp70-BMP interaction by BMP antibodies or a point mutation in Hsp70 (Trp90Phe), as well as the pharmacological and genetic...

  13. Heat shock protein 70 (Hsp70) stimulates proliferation and cytolytic activity of natural killer cells

    NARCIS (Netherlands)

    Multhoff, G; Mizzen, L; Winchester, CC; Milner, CM; Wenk, S; Eissner, G; Kampinga, HH; Laumbacher, B; Johnson, J

    1999-01-01

    We previously demonstrated that lysis of tumor cells that express Hsp70, the highly stress-inducible member of the HSP70 family, on their plasma membrane is mediated by natural killer (NK) cells. Here, we studied the effects of different proteins of the HSP70 family in combination with interleukin 2

  14. Conserved structure and expression of hsp70 paralogs in teleost fishes.

    Science.gov (United States)

    Metzger, David C H; Hemmer-Hansen, Jakob; Schulte, Patricia M

    2016-06-01

    The cytosolic 70KDa heat shock proteins (Hsp70s) are widely used as biomarkers of environmental stress in ecological and toxicological studies in fish. Here we analyze teleost genome sequences to show that two genes encoding inducible hsp70s (hsp70-1 and hsp70-2) are likely present in all teleost fish. Phylogenetic and synteny analyses indicate that hsp70-1 and hsp70-2 are distinct paralogs that originated prior to the diversification of the teleosts. The promoters of both genes contain a TATA box and conserved heat shock elements (HSEs), but unlike mammalian HSP70s, both genes contain an intron in the 5' UTR. The hsp70-2 gene has undergone tandem duplication in several species. In addition, many other teleost genome assemblies have multiple copies of hsp70-2 present on separate, small, genomic scaffolds. To verify that these represent poorly assembled tandem duplicates, we cloned the genomic region surrounding hsp70-2 in Fundulus heteroclitus and showed that the hsp70-2 gene copies that are on separate scaffolds in the genome assembly are arranged as tandem duplicates. Real-time quantitative PCR of F. heteroclitus genomic DNA indicates that four copies of the hsp70-2 gene are likely present in the F. heteroclitus genome. Comparison of expression patterns in F. heteroclitus and Gasterosteus aculeatus demonstrates that hsp70-2 has a higher fold increase than hsp70-1 following heat shock in gill but not in muscle tissue, revealing a conserved difference in expression patterns between isoforms and tissues. These data indicate that ecological and toxicological studies using hsp70 as a biomarker in teleosts should take this complexity into account. PMID:26922644

  15. Tumor imaging and targeting potential of an Hsp70-derived 14-mer peptide.

    Directory of Open Access Journals (Sweden)

    Mathias Gehrmann

    Full Text Available BACKGROUND: We have previously used a unique mouse monoclonal antibody cmHsp70.1 to demonstrate the selective presence of a membrane-bound form of Hsp70 (memHsp70 on a variety of leukemia cells and on single cell suspensions derived from solid tumors of different entities, but not on non-transformed cells or cells from corresponding 'healthy' tissue. This antibody can be used to image tumors in vivo and target them for antibody-dependent cellular cytotoxicity. Tumor-specific expression of memHsp70 therefore has the potential to be exploited for theranostic purposes. Given the advantages of peptides as imaging and targeting agents, this study assessed whether a 14-mer tumor penetrating peptide (TPP; TKDNNLLGRFELSG, the sequence of which is derived from the oligomerization domain of Hsp70 which is expressed on the cell surface of tumor cells, can also be used for targeting membrane Hsp70 positive (memHsp70+ tumor cells, in vitro. METHODOLOGY/PRINCIPAL FINDINGS: The specificity of carboxy-fluorescein (CF- labeled TPP (TPP to Hsp70 was proven in an Hsp70 knockout mammary tumor cell system. TPP specifically binds to different memHsp70+ mouse and human tumor cell lines and is rapidly taken up via endosomes. Two to four-fold higher levels of CF-labeled TPP were detected in MCF7 (82% memHsp70+ and MDA-MB-231 (75% memHsp70+ cells compared to T47D cells (29% memHsp70+ that exhibit a lower Hsp70 membrane positivity. After 90 min incubation, TPP co-localized with mitochondrial membranes in memHsp70+ tumors. Although there was no evidence that any given vesicle population was specifically localized, fluorophore-labeled cmHsp70.1 antibody and TPP preferentially accumulated in the proximity of the adherent surface of cultured cells. These findings suggest a potential association between membrane Hsp70 expression and cytoskeletal elements that are involved in adherence, the establishment of intercellular synapses and/or membrane reorganization. CONCLUSIONS

  16. Role of membrane Hsp70 in radiation sensitivity of tumor cells

    International Nuclear Information System (INIS)

    The major stress-inducible heat shock protein 70 (Hsp70) is frequently overexpressed in the cytosol and integrated in the plasma membrane of tumor cells via lipid anchorage. Following stress such as non-lethal irradiation Hsp70 synthesis is up-regulated. Intracellular located Hsp70 is known to exert cytoprotective properties, however, less is known about membrane (m)Hsp70. Herein, we investigate the role of mHsp70 in the sensitivity towards irradiation in tumor sublines that differ in their cytosolic and/or mHsp70 levels. The isogenic human colon carcinoma sublines CX+ with stable high and CX− with stable low expression of mHsp70 were generated by fluorescence activated cell sorting, the mouse mammary carcinoma sublines 4 T1 (4 T1 ctrl) and Hsp70 knock-down (4 T1 Hsp70 KD) were produced using the CRISPR/Cas9 system, and the Hsp70 down-regulation in human lung carcinoma sublines H1339 ctrl/H1339 HSF-1 KD and EPLC-272H ctrl/EPLC-272H HSF-1 KD was achieved by small interfering (si)RNA against Heat shock factor 1 (HSF-1). Cytosolic and mHsp70 was quantified by Western blot analysis/ELISA and flow cytometry; double strand breaks (DSBs) and apoptosis were measured by flow cytometry using antibodies against γH2AX and real-time PCR (RT-PCR) using primers and antibodies directed against apoptosis related genes; and radiation sensitivity was determined using clonogenic cell surviving assays. CX+/CX− tumor cells exhibited similar cytosolic but differed significantly in their mHsp70 levels, 4 T1 ctrl/4 T1 Hsp70 KD cells showed significant differences in their cytosolic and mHsp70 levels and H1339 ctrl/H1339 HSF-1 KD and EPLC-272H ctrl/EPLC-272H HSF-1 KD lung carcinoma cell sublines had similar mHsp70 but significantly different cytosolic Hsp70 levels. γH2AX was significantly up-regulated in irradiated CX− and 4 T1 Hsp70 KD with low basal mHsp70 levels, but not in their mHsp70 high expressing counterparts, irrespectively of their cytosolic Hsp70 content. After

  17. Reactivation of protein aggregates by mortalin and Tid1--the human mitochondrial Hsp70 chaperone system.

    Science.gov (United States)

    Iosefson, Ohad; Sharon, Shelly; Goloubinoff, Pierre; Azem, Abdussalam

    2012-01-01

    The mitochondrial 70-kDa heat shock protein (mtHsp70), also known in humans as mortalin, is a central component of the mitochondrial protein import motor and plays a key role in the folding of matrix-localized mitochondrial proteins. MtHsp70 is assisted by a member of the 40-kDa heat shock protein co-chaperone family named Tid1 and a nucleotide exchange factor. Whereas, yeast mtHsp70 has been extensively studied in the context of protein import in the mitochondria, and the bacterial 70-kDa heat shock protein was recently shown to act as an ATP-fuelled unfolding enzyme capable of detoxifying stably misfolded polypeptides into harmless natively refolded proteins, little is known about the molecular functions of the human mortalin in protein homeostasis. Here, we developed novel and efficient purification protocols for mortalin and the two spliced versions of Tid1, Tid1-S, and Tid1-L and showed that mortalin can mediate the in vitro ATP-dependent reactivation of stable-preformed heat-denatured model aggregates, with the assistance of Mge1 and either Tid1-L or Tid1-S co-chaperones or yeast Mdj1. Thus, in addition of being a central component of the protein import machinery, human mortalin together with Tid1, may serve as a protein disaggregating machine which, for lack of Hsp100/ClpB disaggregating co-chaperones, may carry alone the scavenging of toxic protein aggregates in stressed, diseased, or aging human mitochondria. PMID:21811887

  18. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard;

    small HSPs). Hsp70 belongs to the HSP70 family and is expressed at low levels in normal non-stressed cells. Its expression is however induced by different cellular stresses, such as heat shock and oxidative stress. The function of Hsp70 depends on its cellular location: Intracellular it has...... normal ER/Golgi transport did not affect Hsp70 surface expression. Intracellular Calcium and the transcription factor Sp1, that has previously been shown to be important for the intracellular stress mediated by HDAC-inhibitors, were not involved in Hsp70 surface expression. We also found that HDAC...... cytoprotective and anti-apoptotic functions, whereas it exerts immunostimulatory functions extracellularly. Secreted Hsp70 is for example involved in cross-presentation of cancer-derived antigenic peptides, a function which is currently explored in immunotherapeutic approaches against cancer. Additionally...

  19. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard; Skov, Søren

    2009-01-01

    We show that inhibition of HDAC activity leads to surface expression of Hsp70 on various hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC inhibitor-mediated Hsp70 surface expression was confined to the apoptotic Annexin V...... activity selectively induces surface expression of Hsp70 on hematopoietic cancer cells and that this may increase immunorecognition of these cells.......-positive cells and blocked by inhibition of apoptosis. Other chemotherapeutic inducers of apoptosis such as etoposide and camptothecin also led to a robust induction of Hsp70 surface expression. Hsp70 expression was, however, not caused by induction of apoptosis per se, as activated CD4 T cells remained Hsp70...

  20. HSP70: a promising target for laryngeal carcinoma radiaotherapy by inhibiting cleavage and degradation of nucleolin

    Directory of Open Access Journals (Sweden)

    Xu Jing

    2010-08-01

    Full Text Available Abstract Previous studies have shown that heat shock proteins (HSPs were upregulated in various types of tumors and were associated with histological grade, recurrence and metastasis of malignant tumors. In this study, we investigated whether heat shock protein 70 kDa (HSP70 was associated with histological grade of laryngeal squamous cell carcinomas (LSCC. We also determine the role of HSP70 in LSCC radiation resistance using a laryngeal carcinoma xenograft model by antisense HSP70 RNA technique. Immunohistochemistry data showed that HSP70 was detected in 96% of LSCC tissues (48 out of 50. The expression level of HSP70 was significantly lower in early stage of LSCC than that in late stage (P = 0.015. Radiation treatment result showed that the volumes and weights of implantation tumors in the group injected with antisense HSP70 oligos were significantly reduced comparing to the group injected with random oligos(p

  1. Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi

    OpenAIRE

    Jolly, Carrie E.; Leonard, Cory A.; J. Russell Hayman

    2010-01-01

    Microsporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 70 family member (Hsp70; ECU02_0100; “C1”) was chosen for further analysis. DNA encoding the C1 hsp70 was amplified, cloned and used to heterologously express the C1 Hsp70 protein, and...

  2. The role of HSP70 in mediating age-dependent mortality in sepsis

    Science.gov (United States)

    McConnell, Kevin W.; Fox, Amy C.; Clark, Andrew T.; Chang, Nai-Yuan Nicholas; Dominguez, Jessica A.; Farris, Alton B.; Buchman, Timothy G.; Hunt, Clayton R.; Coopersmith, Craig M.

    2011-01-01

    Sepsis is primarily a disease of the aged, with increased incidence and mortality occurring in aged hosts. Heat shock protein (HSP) 70 plays an important role in both healthy aging and the stress response to injury. The purpose of this study was to determine the role of HSP70 in mediating mortality and the host inflammatory response in aged septic hosts. Sepsis was induced in both young (6–12week old) and aged (16–17 month old) HSP70−/− and wild type (WT) mice to determine if HSP70 modulated outcome in an age-dependent fashion. Young HSP70−/− and WT mice subjected to cecal ligation and puncture (CLP), Pseudomonas aeruginosa pneumonia or Streptococcus pneumoniae pneumonia had no differences in mortality, suggesting HSP70 does not mediate survival in young septic hosts. In contrast, mortality was higher in aged HSP70−/− mice than aged WT mice subjected to CLP (p=0.01), suggesting HSP70 mediates mortality in sepsis in an age-dependent fashion. Compared to WT mice, aged septic HSP70−/− mice had increased gut epithelial apoptosis and pulmonary inflammation. In addition, HSP70−/−mice had increased systemic levels of TNF-α, IL-6, IL-10 and IL-1β compared to WT mice. These data demonstrate that HSP70 is a key determinant of mortality in aged but not young hosts in sepsis. HSP70 may play a protective role in an age-dependent response to sepsis by preventing excessive gut apoptosis and both pulmonary and systemic inflammation. PMID:21296977

  3. Hsp70 Knockdown by siRNA Decreased Collagen Production in Keloid Fibroblasts

    OpenAIRE

    Shin, Jung U; Lee, Won Jai; Tran, Thanh-Nga; Jung, Inhee; Lee, Ju Hee

    2015-01-01

    Purpose There are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously, we reported that heat shock protein 70 (Hsp70) is up-regulated in keloid fibroblasts and keloid tissue. We, therefore, investigated whether Hsp70 is related to excessive collagen production in keloid fibroblasts. Materials and Methods We inhibited Hsp70 in keloid fibroblasts by RNA interference and examined the resulting collagen expression. Thus, we selected...

  4. Members of the Hsp70 family of proteins in the cell wall of Saccharomyces cerevisiae.

    OpenAIRE

    López-Ribot, J L; Chaffin, W L

    1996-01-01

    Western blot (immunoblot) analysis of cell wall and cytosolic extracts obtained from parental and ssa1 and ssa2 single- and double-mutant strains of Saccharomyces cerevisiae showed that the heat shock protein 70 (Hsp70) products of these genes, previously thought to be restricted to the cell interior, are also present in the cell wall. A cell wall location was further confirmed by indirect immunofluorescence with intact cells and biotinylation of extracellular Hsp70. Hsp70s have been implicat...

  5. Proteomics shows Hsp70 does not bind peptide sequences indiscriminately in vivo

    International Nuclear Information System (INIS)

    Heat shock protein 70 (Hsp70) binds peptide and has several functions that include protein folding, protein trafficking, and involvement with immune function. However, endogenous Hsp70-binding peptides had not previously been identified. Therefore, we eluted and identified several hundred endogenously bound peptides from Hsp70 using liquid chromatography ion trap mass spectrophotometry (LC-ITMS). Our work shows that the peptides are capable of binding Hsp70 as previously described. They are generally 8-26 amino acids in length and correspond to specific regions of many proteins. Through computationally assisted analysis of peptides eluted from Hsp70 we determined variable amino acid sequences, including a 5 amino acid core sequence that Hsp70 favorably binds. We also developed a computer algorithm that predicts Hsp70 binding within proteins. This work helps to define what peptides are bound by Hsp70 in vivo and suggests that Hsp70 facilitates peptide selection by aiding a funneling mechanism that is flexible but allows only a limited number of peptides to be processed

  6. Yeast Two-Hybrid and One-Hybrid Screenings Identify Regulators of hsp70 Gene Expression.

    Science.gov (United States)

    Saito, Youhei; Nakagawa, Takanobu; Kakihana, Ayana; Nakamura, Yoshia; Nabika, Tomomi; Kasai, Michihiro; Takamori, Mai; Yamagishi, Nobuyuki; Kuga, Takahisa; Hatayama, Takumi; Nakayama, Yuji

    2016-09-01

    The mammalian stress protein Hsp105β, which is specifically expressed during mild heat shock and localizes to the nucleus, induces the major stress protein Hsp70. In the present study, we performed yeast two-hybrid and one-hybrid screenings to identify the regulators of Hsp105β-mediated hsp70 gene expression. Six and two proteins were detected as Hsp105β- and hsp70 promoter-binding proteins, respectively. A luciferase reporter gene assay revealed that hsp70 promoter activation is enhanced by the transcriptional co-activator AF9 and splicing mediator SNRPE, but suppressed by the coiled-coil domain-containing protein CCDC127. Of these proteins, the knockdown of SNRPE suppressed the expression of Hsp70 irrespective of the presence of Hsp105β, indicating that SNRPE essentially functions as a transcriptional activator of hsp70 gene expression. The overexpression of HSP70 in tumor cells has been associated with cell survival and drug resistance. We here identified novel regulators of Hsp70 expression in stress signaling and also provided important insights into Hsp70-targeted anti-cancer therapy. J. Cell. Biochem. 117: 2109-2117, 2016. © 2016 Wiley Periodicals, Inc. PMID:26873636

  7. Low resolution structural characterization of the Hsp70-interacting protein - Hip - from Leishmania braziliensis emphasizes its high asymmetry.

    Science.gov (United States)

    Dores-Silva, P R; Silva, E R; Gomes, F E R; Silva, K P; Barbosa, L R S; Borges, J C

    2012-04-15

    The Hsp70 is an essential molecular chaperone in protein metabolism since it acts as a pivot with other molecular chaperone families. Several co-chaperones act as regulators of the Hsp70 action cycle, as for instance Hip (Hsp70-interacting protein). Hip is a tetratricopeptide repeat protein (TPR) that interacts with the ATPase domain in the Hsp70-ADP state, stabilizing it and preventing substrate dissociation. Molecular chaperones from protozoans, which can cause some neglected diseases, are poorly studied in terms of structure and function. Here, we investigated the structural features of Hip from the protozoa Leishmania braziliensis (LbHip), one of the causative agents of the leishmaniasis disease. LbHip was heterologously expressed and purified in the folded state, as attested by circular dichroism and intrinsic fluorescence emission techniques. LbHip forms an elongated dimer, as observed by analytical gel filtration chromatography, analytical ultracentrifugation and small angle X-ray scattering (SAXS). With the SAXS data a low resolution model was reconstructed, which shed light on the structure of this protein, emphasizing its elongated shape and suggesting its domain organization. We also investigated the chemical-induced unfolding behavior of LbHip and two transitions were observed. The first transition was related to the unfolding of the TPR domain of each protomer and the second transition of the dimer dissociation. Altogether, LbHip presents a similar structure to mammalian Hip, despite their low level of conservation, suggesting that this class of eukaryotic protein may use a similar mechanism of action. PMID:22387434

  8. INDUCEMENT OF ANTITUMOR-IMMUNITY BY DC ACTIVATED BY HSP70-H22 TUMOR ANTIGEN PEPTIDE

    Institute of Scientific and Technical Information of China (English)

    冯作化; 黄波; 张桂梅; 李东; 王洪涛

    2003-01-01

    Objective: To investigate the feasibility of decreasing the dosage of tumor antigen peptides by dendritic cell (DC)-presenting and the characteristics of modification of DC by heat shock protein (Hsp70) and antigen peptides. Methods: Peptides were bound to Hsp70 and used to modify DC in vitro. The metabolism of the modified DC and the cytokines secreted by the modified DC were determined. The activation of lymphocytes by the modified DC and Hsp70-H22 peptides was tested. The cytotoxicity of the activated lymphocytes to H22 tumor cells was analyzed. The inhibitory effect of tumor in mice by the injection of DC and Hsp70-H22 peptides was tested. Results: 0.15μg of H22 peptides bound with Hsp70 could make 2×105 DC mature. 4×103 matured DC could activate 2×106 lymphocytes. The same amount of lymphocytes could be activated to produce similar cytotoxicity to tumor cells by either DC modified by 0.003μg of peptides bound with Hsp70 or by direct stimulation with 0.15μg of peptides bound with Hsp70. The dosage of peptides could be reduced by about 50 folds if the modified DC was used for injection instead of Hsp70-peptides. Peptides from normal hepatocytes, bound with Hsp70, could not make DC mature, nor activate lymphocytes through DC. Conclusion: The dosage of Hsp70-H22 peptides can be reduced significantly by DC-presenting to activate lymphocytes. Peptides from normal cells could not activate lymphocytes by either Hsp70-presenting or DC-presenting and they have little chance to induce autoimmunity.

  9. Hsp70 plays an important role in high-fat diet induced gestational hyperglycemia in mice.

    Science.gov (United States)

    Xing, Baoheng; Wang, Lili; Li, Qin; Cao, Yalei; Dong, Xiujuan; Liang, Jun; Wu, Xiaohua

    2015-12-01

    Gestational diabetes mellitus (GDM) has emerged as an epidemic disease during the last decade, affecting about 2 to 5% pregnant women. Even among women who have gestational hyperglycemia may also be positively related to adverse outcomes as GDM. Since heat shock protein (Hsp) 70 has been reported to be associated with diabetes and insulin resistance and its expression was reported to be negatively regulated by the membrane-permeable Hsp70 inhibitor MAL3-101 while positively regulated by the Hsp70 activator BGP-15, we investigated whether Hsp70 played a role in a gestational hyperglycemia mouse model. Mice were divided into non-pregnant and pregnant groups, and each comprised three subgroups: control, high-fat diet (HFD) + MAL3-101, and HFD + BGP-15. We examined the serum levels of triglycerides, total cholesterol, glucose, and insulin, as well as conducted thermal detection of brown adipose tissue (BAT). The role of Hsp70 in BAT apoptosis was also investigated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and caspase-3 staining. Higher serum level of Hsp70 was associated with increased bodyweight gain after pregnancy in mice fed HFD. Circulating Hsp70 was elevated in control pregnant mice compared to control non-pregnant mice. BGP-induced serum Hsp70 expression reduced triglycerides, total cholesterol, glucose, and insulin levels in the serum. Additionally, thermal detection of BAT, TUNEL, and caspase-3 staining revealed relationship correlation between Hsp70 and BAT functions. Hsp70 level is associated with hyperglycemia during pregnancy. Our results support the role of Hsp70 in facilitating BAT activities and protecting BAT cells from apoptosis via caspase-3 pathway. PMID:26318018

  10. Functional analysis of the Hikeshi-like protein and its interaction with HSP70 in Arabidopsis

    Energy Technology Data Exchange (ETDEWEB)

    Koizumi, Shinya; Ohama, Naohiko; Mizoi, Junya [Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan); Shinozaki, Kazuo [RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama, Kanagawa 230-0045 (Japan); Yamaguchi-Shinozaki, Kazuko, E-mail: akys@mail.ecc.u-tokyo.ac.jp [Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan)

    2014-07-18

    Highlights: • HKL, a Hikeshi homologous gene is identified in Arabidopsis. • HKL interacts with two HSP70 isoforms and regulates the subcellular localization of HSC70-1. • The two HSP70 translocate into nucleus in response to heat stress. • Overexpression of HKL confers thermotolerance in transgenic plants. - Abstract: Heat shock proteins (HSPs) refold damaged proteins and are an essential component of the heat shock response. Previously, the 70 kDa heat shock protein (HSP70) has been reported to translocate into the nucleus in a heat-dependent manner in many organisms. In humans, the heat-induced translocation of HSP70 requires the nuclear carrier protein Hikeshi. In the Arabidopsis genome, only one gene encodes a protein with high homology to Hikeshi, and we named this homolog Hikeshi-like (HKL) protein. In this study, we show that two Arabidopsis HSP70 isoforms accumulate in the nucleus in response to heat shock and that HKL interacts with these HSP70s. Our histochemical analysis revealed that HKL is predominantly expressed in meristematic tissues, suggesting the potential importance of HKL during cell division in Arabidopsis. In addition, we show that HKL regulates HSP70 localization, and HKL overexpression conferred thermotolerance to transgenic Arabidopsis plants. Our results suggest that HKL plays a positive role in the thermotolerance of Arabidopsis plants and cooperatively interacts with HSP70.

  11. Inducible heat shock protein 70 (HSP70) Involves in radioadaptive response

    International Nuclear Information System (INIS)

    Radiation induced fibrosarcoma (RIF) cells did not show the adaptive response, i.e. a reduced effect from a higher challenging dose (4 Gy) of a radiation when a priming dose (1 cGy) had been applied 4 or 7 hrs earlier, but its thermoresistant clone (TR) did. Since inducible HSP70 expression was different between two cell lines, the role of inducible HSP70 in adaptive response was examined. When inducible hsp70 was transfected to RIF cells, the adaptive response was acquired. Inducible hsp70 transfected NIH3T3 mouse embryo cells also conferred radioresistance when detected by clonogenic survival, [3H]-thymidine incoporation, and ELISA cell death detection kit, as well as increased tendency of induction adaptive response. Interestingly, basal levels of Ca2+-dependent and Ca2+-independent PKC activities were increased by inducible HSP70, when compared to those of control vector cells. Irradiation of g-ray induced PKC activation within minutes in control vector cells, while it did not by inducible HSP70. Cellular redistribution to particulate fraction of PKCa, d and z by g-rays was not detected either. Furthermore, radioresistance by inducible HSP70 was disappeared by pre-treatment of PKC inhibitors, 1-(5-isoquinolinesulfomyI)-2-methylpepirazine (H7), prolonged treatment of 12-O-tetradecanoylphorbol-13-acetate (TPA), and GF109203X in clonogenic survival assay. Taken together, these data suggest that inducible HSP70-induced radioresistance is associated with its elevated level of PKC activity

  12. Serum levels of HSP70 and other DAMP proteins can aid in patient diagnosis after traumatic injury.

    Science.gov (United States)

    Ren, Biqiong; Zou, Guoying; Huang, Yiran; Xu, Guofeng; Xu, Fei; He, Junyu; Zhu, Haowen; Yu, Ping

    2016-07-01

    Danger-associated molecular patterns (DAMPs) are activated by endogenous signals that originate from stressed, injured, or necrotic cells, signifying "danger" to the host. In this study, we evaluated the expression of the DAMP heat shock protein 70 (HSP70) in trauma patients with and without secondary infections. Levels of glucose (GLU), procalcitonin (PCT), total cholesterol (T-Chol), and white blood cell (WBC) counts were also evaluated at three time stages after trauma. Our analysis showed that the levels of serum HSP70 in patients with minor, moderate, and severe injuries were significantly higher than in healthy patients at each time point post-injury (P < 0.01), and levels of serum HSP70 in the severe injury group were significantly higher than in the minor injury group at 1-6 h after trauma (P = 0.047). HSP70 was correlated with GLU and was negatively correlated with T-Chol in the period 1-6 h after injury (P = 0.008/0.032). WBC and GLU were elevated after trauma, with mutual positive correlation (P < 0.001). PCT levels increased later than WBC counts and GLU levels; these levels were correlated at the two later time periods, 24-48 h and 60-90 h (P = 0.008/0.041). PCT continued to rise in patients with secondary infection, but PCT dropped at the third time period in patients without secondary infection. In summary, our results suggest that danger and stress theory can be used to predict severity of trauma. PMID:27137634

  13. Expression of HSP70 in Peripheral Lymphocytes of the Patients with Allergic Rhinitis

    Institute of Scientific and Technical Information of China (English)

    刘立思; 肖成峰; 张明; 程磊; 王鄂芬; 邬堂春

    2003-01-01

    The expression levels of heat shock protein 70 (HSP70) from peripheral lymphocytes ofthe patients with allergic rhinitis (AR) and the clinical implication were investigated. In the morn-ing, 3 ml of fasting venous blood was taken out. The lymphocytes were isolated by using Ficoll-Hypaque and the expression of HSP70 in the lymphocytes was detected by using Western blot. Inthe AR patients the HSP70 level (41.49± 15.77 integrated optical density, IOD) were significantlyhigher than that in the control group (23.89±10. 13 IOD, P<0.05). Western blot demonstratedthat HSP70 bands in AR patients were more intensive than those in the control group. It was con-cluded that the elevated HSP70 level in peripheral lymphocytes of the AR patients might contributeto the development of AR.

  14. Compound A, a selective glucocorticoid receptor modulator, enhances heat shock protein Hsp70 gene promoter activation.

    Directory of Open Access Journals (Sweden)

    Ilse M Beck

    Full Text Available Compound A possesses glucocorticoid receptor (GR-dependent anti-inflammatory properties. Just like classical GR ligands, Compound A can repress NF-κB-mediated gene expression. However, the monomeric Compound A-activated GR is unable to trigger glucocorticoid response element-regulated gene expression. The heat shock response potently activates heat shock factor 1 (HSF1, upregulates Hsp70, a known GR chaperone, and also modulates various aspects of inflammation. We found that the selective GR modulator Compound A and heat shock trigger similar cellular effects in A549 lung epithelial cells. With regard to their anti-inflammatory mechanism, heat shock and Compound A are both able to reduce TNF-stimulated IκBα degradation and NF-κB p65 nuclear translocation. We established an interaction between Compound A-activated GR and Hsp70, but remarkably, although the presence of the Hsp70 chaperone as such appears pivotal for the Compound A-mediated inflammatory gene repression, subsequent novel Hsp70 protein synthesis is uncoupled from an observed CpdA-induced Hsp70 mRNA upregulation and hence obsolete in mediating CpdA's anti-inflammatory effect. The lack of a Compound A-induced increase in Hsp70 protein levels in A549 cells is not mediated by a rapid proteasomal degradation of Hsp70 or by a Compound A-induced general block on translation. Similar to heat shock, Compound A can upregulate transcription of Hsp70 genes in various cell lines and BALB/c mice. Interestingly, whereas Compound A-dependent Hsp70 promoter activation is GR-dependent but HSF1-independent, heat shock-induced Hsp70 expression alternatively occurs in a GR-independent and HSF1-dependent manner in A549 lung epithelial cells.

  15. Use of an enzyme-linked immunosorbent assay (Elisa) for the determination of HSP 70

    International Nuclear Information System (INIS)

    It has been suggested that HSP 70 is a good indicator to quantify thermotolerance. The authors developed an enzyme-linked immunosorbent assay (Elisa) for measuring the total amount of HSP 70 in mammalian cells. Microtitration plates were incubated with various concentrations of cell lysate in PBS. Milk proteins were used to block non-specific protein binding sites in the wells. Monoclonal antibody against purified HSP 70 raised in a mouse was added, followed by peroxidase conjugated goat-anti-mouse antibody. After washing, the bound peroxidase activity was measured colorimetrically to calculate the original amount of HSP 70 antigen. Kinetic studies indicate that the linear range of the assay corresponds to the amount of HSP 70 from 103-104 HA-1 cells (0.1-2.0 μg cytosolic proteins in each well). Elisa is a highly sensitive method for quantifying HSP 70 and can be easily used to measure the total amount of HSP 70 instead of the synthesis rate of HSP 70 by pulse radiolabeling. The authors are currently developing the sensitive Elisa system for measurement of thermotolerance in human tumors. A comparison of measurement of HSP 70 by pulse labeling the cells with 35S-methionine and by Elisa has been done in HA-1 cells. HA-1 cells were exposed to 450C, 10 min, then incubated at 370C for 0-24 h. These results show that the rate of HSP synthesis increased several fold, although the total amount of HSP 70 is increased less than 2 fold during thermotolerance induction. These results are consistent with data in the literature obtained by other techniques

  16. Hsp70 as an indicator of stress in the cells after contact with nanoparticles

    Science.gov (United States)

    Hardilová, Šárka; Havrdová, Markéta; Panáček, Aleš; Kvítek, Libor; Zbořil, Radek

    2015-05-01

    In recent years, production of nanoparticles is increased and thus grows our contact with them too. Question of safety is closely related to the issue of use nanoparticles. There are a number of tests that monitor the viability, ROS production, the effect on the DNA and cell cycle, however, rarely encountered studies on stress in the cells after contact with nanoparticles. Heat shock proteins (HSP) are among the substances that can be used for monitoring stress in cells. HSP are structures with a chaperone activity. They are evolutionarily very old, conservative and they are found with a high degree of homology in prokaryotes and eukaryotes including humans. They exist at low concentrations under physiological conditions, while in the denaturing conditions e.g. high or low temperature, radiation, exposure to chemicals, heavy metals, or nanoparticles their expression is changed. HSPs are involved in maintaining homeostasis in the cell that the denatured protein conformations allow recovery to the original stage. One of the most common proteins from HSP family is Hsp70 - protein with a molecular weight of 70 kDa. The level of Hsp70 in a cell after exposure to the stress changes depending on the stress level to which the cell is exposed to and a time period during which lasted stressful conditions. Our research monitors stress levels of cells manifesting by Hsp70 production after contact with silver nanoparticles. Nanoparticles show different toxicity towards different types of target cells, which is reflected in the values of IC50 - concentration that kills 50% tested cells. Concentration of test substance toxic to one cell type may be innocuous to cells of another type. IC50 obtained from the MTT assay provides a suitable default data and if multiples of IC50 values are used, we can compare and generalize. Studies can be used to compare stress levels in cells that show different sensitivity to the tested nanoparticles compared with cells under optimal growth

  17. Hsp70 biases the folding pathways of client proteins.

    Science.gov (United States)

    Sekhar, Ashok; Rosenzweig, Rina; Bouvignies, Guillaume; Kay, Lewis E

    2016-05-17

    The 70-kDa heat shock protein (Hsp70) family of chaperones bind cognate substrates to perform a variety of different processes that are integral to cellular homeostasis. Although detailed structural information is available on the chaperone, the structural features of folding competent substrates in the bound form have not been well characterized. Here we use paramagnetic relaxation enhancement (PRE) NMR spectroscopy to probe the existence of long-range interactions in one such folding competent substrate, human telomere repeat binding factor (hTRF1), which is bound to DnaK in a globally unfolded conformation. We show that DnaK binding modifies the energy landscape of the substrate by removing long-range interactions that are otherwise present in the unbound, unfolded conformation of hTRF1. Because the unfolded state of hTRF1 is only marginally populated and transiently formed, it is inaccessible to standard NMR approaches. We therefore developed a (1)H-based CEST experiment that allows measurement of PREs in sparse states, reporting on transiently sampled conformations. Our results suggest that DnaK binding can significantly bias the folding pathway of client substrates such that secondary structure forms first, followed by the development of longer-range contacts between more distal parts of the protein. PMID:27140645

  18. Increased hsp70 of glucocorticoid receptor complex induced by scald and heat stress and its possible effect on the affinity of glucocorticoid receptor

    Institute of Scientific and Technical Information of China (English)

    WANG Xiao-hui; TANG Hong-tai; LU Jian; XIA Zhao-fan

    2010-01-01

    Background Glucocorticoid (GC) insensitivity/GC resistance is an important etiological and prognostic factor in multiple diseases and pathophysiological processes such as scald, shock and asthma. The function of GC was mediated by glucocorticoid receptor (GR). Scald not only decreased the expression of GR but also reduced the affinity of GR, which played an important role in GC resistance in scalded rats. Whereas the molecular mechanism responsible for the decrease of GR affinity resulted from scald remains unclear. Recent studies showed that the changes of heat shock proteins (hsp) especially hsp90 and hsp70 of GR heterocomplex were associated with GR low affinity in vitro. Methods The affinity of GR in hepatic cytosols and in the cytosols of SMMC-7721 cells were determined by radioligand binding assay and scatchard plot. GR heterocomplex in cytosols were captured by coimmunoprecipation and the levels of hsp90 and hsp70 of GR complex were detected by quantitative Western blotting.Results Similar with that of hepatic cytosol of scalded rats, a remarkable decrease of GR affinity was also found in the cytosol of heat stressed SMMC-7721 cells. The level of hsp70 of GR complex in hepatic cytosol of scalded rats (30% total body surface area immersion scald) and in cytosol of heat stressed human hepatocarcinoma cell line SMMC-7721 were both increased by 1.5 fold, whereas no change of hsp90 in GR heterocomplex was found. According to the correlation analysis, there may be a positive relationship between increased hsp70 of GR complex and decreased GR affinity in the cytosols.Conclusions The primary results indicated that the level of hsp70 of GR heterocomplex was increased in the hepatic cytosol of scalded rats and the cytosol of heat stressed SMMC-7721 cells. The increase of hsp70 of GR complex might be associated with the decrease of GR affinity.

  19. Enhanced Hsp70 expression protects against acute lung injury by modulating apoptotic pathways.

    Directory of Open Access Journals (Sweden)

    Gabriella Aschkenasy

    Full Text Available The Acute respiratory distress syndrome (ARDS is a highly lethal inflammatory lung disorder. Apoptosis plays a key role in its pathogenesis. We showed that an adenovirus expressing the 70 kDa heat shock protein Hsp70 (AdHSP protected against sepsis-induced lung injury. In this study we tested the hypothesis that AdHSP attenuates apoptosis in sepsis-induced lung injury. Sepsis was induced in rats via cecal ligation and double puncture (2CLP. At the time of 2CLP PBS, AdHSP or AdGFP (an adenoviral vector expressing green fluorescent protein were injected into the tracheas of septic rats. 48 hours later, lungs were isolated. One lung was fixed for TUNEL staining and immunohistochemistry. The other was homogenized to isolate cytosolic and nuclear protein. Immunoblotting, gel filtration and co-immunoprecipitation were performed in these extracts. In separate experiments MLE-12 cells were incubated with medium, AdHSP or AdGFP. Cells were stimulated with TNFα. Cytosolic and nuclear proteins were isolated. These were subjected to immunoblotting, co-immunoprecipitation and a caspase-3 activity assay. TUNEL assay demonstrated that AdHSP reduced alveolar cell apoptosis. This was confirmed by immunohistochemical detection of caspase 3 abundance. In lung isolated from septic animals, immunoblotting, co-immunoprecipitation and gel filtration studies revealed an increase in cytoplasmic complexes containing caspases 3, 8 and 9. AdHSP disrupted these complexes. We propose that Hsp70 impairs apoptotic cellular pathways via interactions with caspases. Disruption of large complexes resulted in stabilization of lower molecular weight complexes, thereby, reducing nuclear caspase-3. Prevention of apoptosis in lung injury may preserve alveolar cells and aid in recovery.

  20. Expression and Significance of HSP70 and MGMT in Breast Neoplasms%HSP70和MGMT在乳腺肿瘤中的表达和意义

    Institute of Scientific and Technical Information of China (English)

    吴荣薇; 薄爱华; 王发亮; 芦广萍

    2008-01-01

    目的:探讨乳腺肿瘤组织中热休克蛋白HSP70和DNA修复酶O6-甲基鸟嘌呤-DNA甲基转移酶(O6-methyguanine-DNA methytransferase,MGMT)的表达及临床意义.方法:采用免疫组织化学SABC法检测65例乳腺肿瘤组织中HSP70和MGMT的表达情况.结果:乳腺肿瘤中HSP70的阳性率为80.00%(52/65),MGMT的阳性率为60.00%(39/65),二者呈中度相关性(r=0.46).其中,有淋巴结转移组的阳性率高于无转移组(P<0.05).结论:HSP70和MGMT的表达率与淋巴结转移情况有关.检测HSP70与MGMT对乳腺肿瘤预后和化疗方案的选择有指导意义.

  1. Use of a Chimeric Hsp70 to Enhance the Quality of Recombinant Plasmodium falciparum S-Adenosylmethionine Decarboxylase Protein Produced in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Xolani Henry Makhoba

    Full Text Available S-adenosylmethionine decarboxylase (PfAdoMetDC from Plasmodium falciparum is a prospective antimalarial drug target. The production of recombinant PfAdoMetDC for biochemical validation as a drug target is important. The production of PfAdoMetDC in Escherichia coli has been reported to result in unsatisfactory yields and poor quality product. The co-expression of recombinant proteins with molecular chaperones has been proposed as one way to improve the production of the former in E. coli. E. coli heat shock proteins DnaK, GroEL-GroES and DnaJ have previously been used to enhance production of some recombinant proteins. However, the outcomes were inconsistent. An Hsp70 chimeric protein, KPf, which is made up of the ATPase domain of E. coli DnaK and the substrate binding domain of P. falciparum Hsp70 (PfHsp70 has been previously shown to exhibit chaperone function when it was expressed in E. coli cells whose resident Hsp70 (DnaK function was impaired. We proposed that because of its domain constitution, KPf would most likely be recognised by E. coli Hsp70 co-chaperones. Furthermore, because it possesses a substrate binding domain of plasmodial origin, KPf would be primed to recognise recombinant PfAdoMetDC expressed in E. coli. First, using site-directed mutagenesis, followed by complementation assays, we established that KPf with a mutation in the hydrophobic residue located in its substrate binding cavity was functionally compromised. We further co-expressed PfAdoMetDC with KPf, PfHsp70 and DnaK in E. coli cells either in the absence or presence of over-expressed GroEL-GroES chaperonin. The folded and functional status of the produced PfAdoMetDC was assessed using limited proteolysis and enzyme assays. PfAdoMetDC co-expressed with KPf and PfHsp70 exhibited improved activity compared to protein co-expressed with over-expressed DnaK. Our findings suggest that chimeric KPf may be an ideal Hsp70 co-expression partner for the production of recombinant

  2. Stress-induced interaction between p38 MAPK and HSP70

    International Nuclear Information System (INIS)

    Highlights: ► HSP70 interacts to p38 MAPK in vitro and in vivo. ► HSP70 co-localizes with p38 MAPK in the nucleus upon stress. ► HSP70 is involved in the nuclear phosphorylation of MK2 by p38 MAPK. -- Abstract: p38 MAPK, one of the four MAPK subfamilies in mammalian cells, is activated by environmental stresses and pro-inflammatory cytokines, playing fundamental roles in many biological processes. Despite all that is known on the structure and functions of p38, many questions still exist. The coupling of activation and nuclear translocation represents an important aspect of p38 signaling. In our effort in exploring the potential chaperone for p38 translocation, we performed an endogenous pull-down assay and identified HSP70 as a potential interacting protein of p38. We confirmed the interaction between p38 and HSP70 in vitro and in vivo, and identified their interaction domains. We also showed stress-induced nuclear co-localization of these two proteins. Our preliminary result indicated that HSP70 was related to the phosphorylation of MK2, a specific nuclear downstream target of p38, suggesting HSP70 is a potential chaperone for the nuclear translocation of p38.

  3. A mathematical model of the hsp70 regulation in the cell.

    Science.gov (United States)

    Peper, A; Grimbergen, C A; Spaan, J A; Souren, J E; van Wijk, R

    1998-01-01

    A mathematical model of the regulation process of the heat shock protein hsp70 in the cell is presented. The model describes the damaging effect of elevated temperature on proteins; the interaction of free hsp70 with injured proteins and its chaperone role in nascent protein translation; the relation between the amount of free hsp70 and the formation of the activated trimer form of the heat shock factor protein (HSF); the binding of activated HSF with the heat shock elements on the DNA; the transcription of mRNA of hsp70 and the synthesis of hsp70. The reaction of the model to a temporal rise in temperature shows an initial decline and a subsequent sharp rise to an ultimately increased level of free hsp70 in the cell. The response of the model to both a single and two consecutive heat shocks appears to closely resemble experimental data on hsp70 synthesis. This general agreement demonstrates the structure of the model to be sound and suitable as a basis for further modelling the complex tolerance mechanism of the cell. PMID:9483450

  4. The Effects of Venlafaxine and Dexamethasone on the Expression of HSP70 in Rat C6 Glioma Cells

    OpenAIRE

    Yu, Jaehak; Roh, Sungwon; Lee, Jun-Seok; Yang, Byung-Hwan; Choi, Mi Ran; Chai, Young Gyu; Kim, Seok Hyeon

    2010-01-01

    Objective The present study aimed to determine the intracellular action of the antidepressant, venlafaxine, in C6 glioma cells using heat shock protein 70 (HSP70) immunocytochemistry and HSP70 Western blots; HSP70 is known to be associated with stress and depression. Methods The extent of HSP70 expression was measured after rat C6 glioma cells were treated with 1) dexamethasone only, 2) venlafaxine only, 3) simultaneous venlafaxine and dexamethasone, or 4) dexamethasone after venlafaxine pret...

  5. Nitric oxide and HSP70 proteins during normal pregnancy, gestosis, and preclinical gestosis.

    Science.gov (United States)

    Bloshchinskaya, I A; Davidovich, I M

    2003-03-01

    We measured the content of NO metabolites in blood plasma and urine and concentration of HSP70 proteins in the plasma and leukocytes from 126 women with normal pregnancy, gestosis, and preclinical gestosis. In women with gestosis accompanied by NO deficiency the concentration of HSP70 proteins in the plasma increased more significantly than in leukocytes. These changes are an important marker of endothelial dysfunction and reflect the severity of cell damage during this disorder. The decrease in excretion of NO in the urine and increase in HSP70 protein content in leukocytes and plasma characterize endothelial dysfunction in women with preclinical gestosis. PMID:12802391

  6. Changing of the HSP70 Content in the Baikal Endemic Sponges Lubomirskiidae Under Conditions of Hyperthermia

    Directory of Open Access Journals (Sweden)

    Itskovich V.B.

    2015-12-01

    Full Text Available Baikal endemic sponges (Lubomirskiidae make up the bulk of the benthos biomass of the lake. For the first time the changes in the content of HSP70 in response to elevated environment temperature were analyzed in three endemic species of Baikal sponges: Baikalospongia bacillifera (Dybowski, 1880, B. intermedia (Dybowski, 1880 and Swartschewskia papyracea (Dybowski, 1880. Interspecific variability of constitutive HSP70 level was revealed for representatives of the three analyzed Lubomirskiidae species. After exposure at 13 °С for 3 and 7 days opposite changes were noted in the amount of HSP70. Under conditions of hyperthermia the protein level decrease at Baikalospongia species, while at the S. papyracea HSP70 content slightly increased. The differences in the mechanisms of stress adaptation probably affect the thermal resistance of the species, as well as are evidence supporting their specific status.

  7. Genetics of human longevity with emphasis on the relevance of HSP70 as candidate genes

    DEFF Research Database (Denmark)

    Singh, Ripudaman; Kølvrå, Steen; Rattan, Suresh I S

    2007-01-01

    mechanisms. One such pathway includes the battery of stress response genes, especially the heat shock protein HSP70 genes. Three such genes, HSPA1A, HSPA1B and HSPA1L, are present within the MHC-III region on the short arm of chromosome 6. We and others have found alleles, genotypes and haplotypes which have...... heat shock. Stress response genes, particularly HSP70, are now the major candidates in the gene-longevity association studies....

  8. Using Lymphocyte and Plasma Hsp70 as Biomarkers for Assessing Coke Oven Exposure among Steel Workers

    OpenAIRE

    Yang, Xiaobo; Zheng, Jinping; Bai, Yun; Tian, Fengjie; Yuan, Jing; Sun, Jianya; Liang, Huashan; Guo, Liang; Tan, Hao; Chen, Weihong; Tanguay, Robert M.; Wu, Tangchun

    2007-01-01

    Background Hsp70, an early-response protein induced when organisms are confronted with simple or complicated environmental stresses, can act as either a cellular protector or a danger signal. Objectives The goal of this study was to evaluate levels of lymphocyte and/or plasma Hsp70 as biomarkers for assessing exposure response to complex coke oven emissions (COEs). Methods We recruited 101 coke oven workers and determined levels of polycyclic aromatic hydrocarbon (PAH) exposure, urinary 1-hyd...

  9. Serum level of soluble Hsp70 is associated with vascular calcification

    OpenAIRE

    Krepuska, Miklós; Szeberin, Zoltán; Sótonyi, Péter; Sarkadi, Hunor; Fehérvári, Mátyás; Apor, Astrid; Rimely, Endre; Prohászka, Zoltán; Acsády, György

    2010-01-01

    It has been previously reported that serum levels of 70-kDa heat shock protein (Hsp70) are elevated in peripheral artery disease. The aim of the present study was to examine whether increased serum Hsp70 levels are related to the extent of arterial calcification and standard laboratory parameters of patients with peripheral artery disease, as well as to markers of inflammation (C-reactive protein), atherosclerosis (homocysteine), and calcification (fetuin-a). One hundred eighty chronic athero...

  10. Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae.

    OpenAIRE

    Slater, M R; Craig, E A

    1987-01-01

    The yeast Saccharomyces cerevisiae contains three heat-inducible hsp70 genes. We have characterized the promoter region of the hsp70 heat shock gene YG100, that also displays a basal level of expression. Deletion of the distal region of the promoter resulted in an 80% drop in the basal level of expression without affecting expression after heat shock. Progressive-deletion analysis suggested that sequences necessary for heat-inducible expression are more proximal, within 233 base pairs of the ...

  11. The effect of glutamine infusion on the inflammatory response and HSP70 during human experimental endotoxaemia

    OpenAIRE

    Andreasen, Anne Sofie; Pedersen-Skovsgaard, Theis; Mortensen, Ole Hartvig; van Hall, Gerrit; Moseley, Pope Lloyd; Pedersen, Bente Klarlund

    2009-01-01

    Introduction Glutamine supplementation has beneficial effects on morbidity and mortality in critically ill patients, possibly in part through an attenuation of the proinflammatory cytokine response and a stimulation of heat shock protein (HSP)70. We infused either alanine-glutamine or saline during endotoxin challenge and measured plasma cytokines and HSP70 protein expression. Methods This crossover study, conducted in eight healthy young men, was double-blind, randomized and placebo-controll...

  12. HSP70 overexpression may play a protective role in the mouse embryos stimulated by CUMS

    OpenAIRE

    Li, Xiao-Hong; Pang, Hou-Qing; QIN, LANG; Jin, Song; Zeng, Xun; Bai, Yu; Li, Shang-Wei

    2015-01-01

    Background We evaluated whether heat shock protein HSP70 plays a protective role in the embryos of Kunming mice subjected to chronic unpredictable mild stress. Methods Female mice were stimulated for 4 weeks with nine stressors and then divided into mild, moderate and severe stress groups. Superovulation was induced with a gonadotropin preparation (PMSG/HCG) and HSP70 expression in 2-cell embryos and day 4 embryos was detected by immunofluorescence (IF) and real-time polymerase chain reaction...

  13. Effect of betaine on HSP70 expression and cell survival during adaptation to osmotic stress.

    Science.gov (United States)

    Petronini, P G; De Angelis, E M; Borghetti, A F; Wheeler, K P

    1993-07-15

    Induced expression of the HSP70 gene in 3T3 and SV-3T3 cells was monitored by measurements of the synthesis of HSP70 and of the cellular contents of both HSP70 and its mRNA. The presence of betaine (N-trimethylglycine) at concentrations of 2.5-25 mM decreased the induction of HSP70 gene expression caused by incubation of 3T3 and SV-3T3 cells in hypertonic (0.5 osM) medium. This effect was accompanied by an enhancement of SV-3T3 cell adaptation, assayed by colony formation, to the hyperosmotic conditions. In contrast, the presence of betaine did not affect HSP70 gene expression induced in these cells by heat shock. After 6 h incubation with 25 mM betaine under hypertonic (0.5 osM) conditions the intracellular concentration of betaine in SV-3T3 cells was about 195 mM, compared with about 70 mM under isotonic (0.3 osM) conditions. Hence, with this concentration of extracellular betaine, the marked increase in the accumulation of betaine within the cells presumably counteracts the imposed osmotic pressure and eliminates the signal that otherwise initiates increased expression of the HSP70 gene. PMID:8343134

  14. Maintenance of structure and function of mitochondrial Hsp70 chaperones requires the chaperone Hep1

    Science.gov (United States)

    Sichting, Martin; Mokranjac, Dejana; Azem, Abdussalam; Neupert, Walter; Hell, Kai

    2005-01-01

    Hsp70 chaperones mediate folding of proteins and prevent their misfolding and aggregation. We report here on a new kind of Hsp70 interacting protein in mitochondria, Hep1. Hep1 is a highly conserved protein present in virtually all eukaryotes. Deletion of HEP1 results in a severe growth defect. Cells lacking Hep1 are deficient in processes that need the function of mitochondrial Hsp70s, such as preprotein import and biogenesis of proteins containing FeS clusters. In the mitochondria of these cells, Hsp70s, Ssc1 and Ssq1 accumulate as insoluble aggregates. We show that it is the nucleotide-free form of mtHsp70 that has a high tendency to self-aggregate. This process is efficiently counteracted by Hep1. We conclude that Hep1 acts as a chaperone that is necessary and sufficient to prevent self-aggregation and to thereby maintain the function of the mitochondrial Hsp70 chaperones. PMID:15719019

  15. Variation in Hsp70-1A Expression Contributes to Skin Color Diversity.

    Science.gov (United States)

    Murase, Daiki; Hachiya, Akira; Fullenkamp, Rachel; Beck, Anita; Moriwaki, Shigeru; Hase, Tadashi; Takema, Yoshinori; Manga, Prashiela

    2016-08-01

    The wide range in human skin color results from varying levels of the pigment melanin. Genetic mechanisms underlying coloration differences have been explored, but identified genes do not account for all variation seen in the skin color spectrum. Post-transcriptional and post-translational regulation of factors that determine skin color, including melanin synthesis in epidermal melanocytes, melanosome transfer to keratinocytes, and melanosome degradation, is also critical for pigmentation. We therefore investigated proteins that are differentially expressed in melanocytes derived from either white or African American skin. Two-dimensional gel electrophoresis and mass spectrometry demonstrated that heat shock protein 70-1A (Hsp70-1A) protein levels were significantly higher in African American melanocytes compared with white melanocytes. Hsp70-1A expression significantly correlated with levels of tyrosinase, the rate-limiting melanogenic enzyme, consistent with a proposed role for Hsp70 family members in tyrosinase post-translational modification. In addition, pharmacologic inhibition and small interfering RNA-mediated downregulation of Hsp70-1A correlated with pigmentation changes in cultured melanocytes, modified human skin substitutes, and ex vivo skin. Furthermore, Hsp70-1A inhibition led to increased autophagy-mediated melanosome degradation in keratinocytes. Our data thus reveal that epidermal Hsp70-1A contributes to the diversity of skin color by regulating the amount of melanin synthesized in melanocytes and modulating autophagic melanosome degradation in keratinocytes. PMID:27094592

  16. Identification of an allosteric pocket on human hsp70 reveals a mode of inhibition of this therapeutically important protein.

    Science.gov (United States)

    Rodina, Anna; Patel, Pallav D; Kang, Yanlong; Patel, Yogita; Baaklini, Imad; Wong, Michael J H; Taldone, Tony; Yan, Pengrong; Yang, Chenghua; Maharaj, Ronnie; Gozman, Alexander; Patel, Maulik R; Patel, Hardik J; Chirico, William; Erdjument-Bromage, Hediye; Talele, Tanaji T; Young, Jason C; Chiosis, Gabriela

    2013-12-19

    Hsp70s are important cancer chaperones that act upstream of Hsp90 and exhibit independent anti-apoptotic activities. To develop chemical tools for the study of human Hsp70, we developed a homology model that unveils a previously unknown allosteric site located in the nucleotide binding domain of Hsp70. Combining structure-based design and phenotypic testing, we discovered a previously unknown inhibitor of this site, YK5. In cancer cells, this compound is a potent and selective binder of the cytosolic but not the organellar human Hsp70s and has biological activity partly by interfering with the formation of active oncogenic Hsp70/Hsp90/client protein complexes. YK5 is a small molecule inhibitor rationally designed to interact with an allosteric pocket of Hsp70 and represents a previously unknown chemical tool to investigate cellular mechanisms associated with Hsp70. PMID:24239008

  17. Induction of heat shock protein 70 (Hsp70 prevents neuregulin-induced demyelination by enhancing the proteasomal clearance of c-Jun

    Directory of Open Access Journals (Sweden)

    Rick T Dobrowsky

    2012-12-01

    Full Text Available Modulating molecular chaperones is emerging as an attractive approach to treat neurodegenerative diseases associated with protein aggregation, DPN (diabetic peripheral neuropathy and possibly, demyelinating neuropathies. KU-32 [N-(7-((2R,3R,4S,5R-3,4-dihydroxy-5-methoxy-6,6-dimethyl-tetrahydro-2H-pyran-2-yloxy-8-methyl-2-oxo-2H-chromen-3-ylacetamide] is a small molecule inhibitor of Hsp90 (heat shock protein 90 and reverses sensory deficits associated with myelinated fibre dysfunction in DPN. Additionally, KU-32 prevented the loss of myelinated internodes induced by treating myelinated SC (Schwann cell-DRG (dorsal root ganglia sensory neuron co-cultures with NRG1 (neuregulin-1 Type 1. Since KU-32 decreased NRG1-induced demyelination in an Hsp70-dependent manner, the goal of the current study was to clarify how Hsp70 may be mechanistically linked to preventing demyelination. The activation of p42/p44 MAPK (mitogen-activated protein kinase and induction of the transcription factor c-Jun serve as negative regulators of myelination. NRG1 activated MAPK, induced c-Jun expression and promoted a loss of myelin segments in DRG explants isolated from both WT (wild-type and Hsp70 KO (knockout mice. Although KU-32 did not block the activation of MAPK, it blocked c-Jun induction and protected against a loss of myelinated segments in WT mice. In contrast, KU-32 did not prevent the NRG1-dependent induction of c-Jun and loss of myelin segments in explants from Hsp70 KO mice. Overexpression of Hsp70 in myelinated DRG explants prepared from WT or Hsp70 KO mice was sufficient to block the induction of c-Jun and the loss of myelin segments induced by NRG1. Lastly, inhibiting the proteasome prevented KU-32 from decreasing c-Jun levels. Collectively, these data support that Hsp70 induction is sufficient to prevent NRG1-induced demyelination by enhancing the proteasomal degradation of c-Jun.

  18. Ionizing radiation improves glioma-specific targeting of superparamagnetic iron oxide nanoparticles conjugated with cmHsp70.1 monoclonal antibodies (SPION-cmHsp70.1)

    Science.gov (United States)

    Shevtsov, Maxim A.; Nikolaev, Boris P.; Ryzhov, Vyacheslav A.; Yakovleva, Ludmila Y.; Marchenko, Yaroslav Y.; Parr, Marina A.; Rolich, Valerij I.; Mikhrina, Anastasiya L.; Dobrodumov, Anatolii V.; Pitkin, Emil; Multhoff, Gabriele

    2015-12-01

    The stress-inducible 72 kDa heat shock protein Hsp70 is known to be expressed on the membrane of highly aggressive tumor cells including high-grade gliomas, but not on the corresponding normal cells. Membrane Hsp70 (mHsp70) is rapidly internalized into tumor cells and thus targeting of mHsp70 might provide a promising strategy for theranostics. Superparamagnetic iron oxide nanoparticles (SPIONs) are contrast negative agents that are used for the detection of tumors with MRI. Herein, we conjugated the Hsp70-specific antibody (cmHsp70.1) which is known to recognize mHsp70 to superparamagnetic iron nanoparticles to assess tumor-specific targeting before and after ionizing irradiation. In vitro experiments demonstrated the selectivity of SPION-cmHsp70.1 conjugates to free and mHsp70 in different tumor cell types (C6 glioblastoma, K562 leukemia, HeLa cervix carcinoma) in a dose-dependent manner. High-resolution MRI (11 T) on T2-weighted images showed the retention of the conjugates in the C6 glioma model. Accumulation of SPION-cmHsp70.1 nanoparticles in the glioma resulted in a nearly 2-fold drop of values in comparison to non-conjugated SPIONs. Biodistribution analysis using NLR-M2 measurements showed a 7-fold increase in the tumor-to-background (normal brain) uptake ratio of SPION-cmHsp70.1 conjugates in glioma-bearing rats in comparison to SPIONs. This accumulation within Hsp70-positive glioma was further enhanced after a single dose (10 Gy) of ionizing radiation. Elevated accumulation of the magnetic conjugates in the tumor due to radiosensitization proves the combination of radiotherapy and application of Hsp70-targeted agents in brain tumors.The stress-inducible 72 kDa heat shock protein Hsp70 is known to be expressed on the membrane of highly aggressive tumor cells including high-grade gliomas, but not on the corresponding normal cells. Membrane Hsp70 (mHsp70) is rapidly internalized into tumor cells and thus targeting of mHsp70 might provide a promising strategy

  19. Short-duration-focused ultrasound stimulation of Hsp70 expression in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Kruse, D E; Ferrara, K W [Department of Biomedical Engineering, University of California, Davis, CA 95616 (United States); Mackanos, M A; O' Connell-Rodwell, C E; Contag, C H [Stanford University School of Medicine, Stanford, CA 94305 (United States)], E-mail: dekruse@ucdavis.edu, E-mail: kwferrara@ucdavis.edu, E-mail: mackanos@stanford.edu, E-mail: ceoconnell@stanford.edu, E-mail: ccontag@stanford.edu

    2008-07-07

    The development of transgenic reporter mice and advances in in vivo optical imaging have created unique opportunities to assess and analyze biological responses to thermal therapy directly in living tissues. Reporter mice incorporating the regulatory regions from the genes encoding the 70 kDa heat-shock proteins (Hsp70) and firefly luciferase (luc) as reporter genes can be used to non-invasively reveal gene activation in living tissues in response to thermal stress. High-intensity-focused ultrasound (HIFU) can deliver measured doses of acoustic energy to highly localized regions of tissue at intensities that are sufficient to stimulate Hsp70 expression. We report activation of Hsp70-luc expression using 1 s duration HIFU heating to stimulate gene expression in the skin of the transgenic reporter mouse. Hsp70 expression was tracked for 96 h following the application of 1.5 MHz continuous-wave ultrasound with spatial peak intensities ranging from 53 W cm{sup -2} up to 352 W cm{sup -2}. The results indicated that peak Hsp70 expression is observed 6-48 h post-heating, with significant activity remaining at 96 h. Exposure durations were simulated using a finite-element model, and the predicted temperatures were found to be consistent with the observed Hsp70 expression patterns. Histological evaluation revealed that the thermal damage starts at the stratum corneum and extends deeper with increasing intensity. These results indicated that short-duration HIFU may be useful for inducing heat-shock expression, and that the period between treatments needs to be greater than 96 h due to the protective properties of Hsp70.

  20. A Surveillance Function of the HSPB8-BAG3-HSP70 Chaperone Complex Ensures Stress Granule Integrity and Dynamism.

    Science.gov (United States)

    Ganassi, Massimo; Mateju, Daniel; Bigi, Ilaria; Mediani, Laura; Poser, Ina; Lee, Hyun O; Seguin, Samuel J; Morelli, Federica F; Vinet, Jonathan; Leo, Giuseppina; Pansarasa, Orietta; Cereda, Cristina; Poletti, Angelo; Alberti, Simon; Carra, Serena

    2016-09-01

    Stress granules (SGs) are ribonucleoprotein complexes induced by stress. They sequester mRNAs and disassemble when the stress subsides, allowing translation restoration. In amyotrophic lateral sclerosis (ALS), aberrant SGs cannot disassemble and therefore accumulate and are degraded by autophagy. However, the molecular events causing aberrant SG formation and the molecular players regulating this transition are largely unknown. We report that defective ribosomal products (DRiPs) accumulate in SGs and promote a transition into an aberrant state that renders SGs resistant to RNase. We show that only a minor fraction of aberrant SGs is targeted by autophagy, whereas the majority disassembles in a process that requires assistance by the HSPB8-BAG3-HSP70 chaperone complex. We further demonstrate that HSPB8-BAG3-HSP70 ensures the functionality of SGs and restores proteostasis by targeting DRiPs for degradation. We propose a system of chaperone-mediated SG surveillance, or granulostasis, which regulates SG composition and dynamics and thus may play an important role in ALS. PMID:27570075

  1. DNA double strand breaks and Hsp70 expression in proton irradiated living cells

    Energy Technology Data Exchange (ETDEWEB)

    Fiedler, Anja [Institute for Experimental Physics II, University of Leipzig (Germany) and Faculty of Biology, Pharmacy and Psychology, University of Leipzig (Germany)]. E-mail: afiedler@uni-leipzig.de; Reinert, Tilo [Institute for Experimental Physics II, University of Leipzig (Germany); Tanner, Judith [Clinic and Polyclinic for Radiation Oncology, University of Halle-Wittenberg (Germany); Butz, Tilman [Institute for Experimental Physics II, University of Leipzig (Germany)

    2007-07-15

    DNA double strand breaks (DSBs) in living cells can be directly provoked by ionising radiation. DSBs can be visualized by immunostaining the phosphorylated histone {gamma}H2AX. Our concern was to test the feasibility of {gamma}H2AX staining for a direct visualization of single proton hits. If single protons produce detectable foci, DNA DSBs could be used as 'biological track detectors' for protons. Ionising radiation can also damage proteins indirectly by inducing free radicals. Heat shock proteins (Hsp) help to refold or even degrade the damaged proteins. The level of the most famous heat shock protein Hsp70 is increased by ionising radiation. We investigated the expression of {gamma}H2AX and Hsp70 after cross and line patterned irradiation with counted numbers of 2.25 MeV protons on primary human skin fibroblasts. The proton induced DSBs appear more delocalised than it was expected by the ion hit accuracy. Cooling the cells before the irradiation reduces the delocalisation of DNA DSBs, which is probably caused by the reduced diffusion of DNA damaging agents. Proton irradiation seems to provoke protein damages mainly in the cytoplasm indicated by cytoplasmic Hsp70 aggregates. On the contrary, in control heat shocked cells the Hsp70 was predominantly localized in the cell nucleus. However, the irradiated area could not be recognized, all cells on the Si{sub 3}N{sub 4} window showed a homogenous Hsp70 expression pattern.

  2. Defining Hsp70 Subnetworks in Dengue Virus Replication Reveals Key Vulnerability in Flavivirus Infection.

    Science.gov (United States)

    Taguwa, Shuhei; Maringer, Kevin; Li, Xiaokai; Bernal-Rubio, Dabeiba; Rauch, Jennifer N; Gestwicki, Jason E; Andino, Raul; Fernandez-Sesma, Ana; Frydman, Judith

    2015-11-19

    Viral protein homeostasis depends entirely on the machinery of the infected cell. Accordingly, viruses can illuminate the interplay between cellular proteostasis components and their distinct substrates. Here, we define how the Hsp70 chaperone network mediates the dengue virus life cycle. Cytosolic Hsp70 isoforms are required at distinct steps of the viral cycle, including entry, RNA replication, and virion biogenesis. Hsp70 function at each step is specified by nine distinct DNAJ cofactors. Of these, DnaJB11 relocalizes to virus-induced replication complexes to promote RNA synthesis, while DnaJB6 associates with capsid protein and facilitates virion biogenesis. Importantly, an allosteric Hsp70 inhibitor, JG40, potently blocks infection of different dengue serotypes in human primary blood cells without eliciting viral resistance or exerting toxicity to the host cells. JG40 also blocks replication of other medically-important flaviviruses including yellow fever, West Nile and Japanese encephalitis viruses. Thus, targeting host Hsp70 subnetworks provides a path for broad-spectrum antivirals. PMID:26582131

  3. DNA double strand breaks and Hsp70 expression in proton irradiated living cells

    International Nuclear Information System (INIS)

    DNA double strand breaks (DSBs) in living cells can be directly provoked by ionising radiation. DSBs can be visualized by immunostaining the phosphorylated histone γH2AX. Our concern was to test the feasibility of γH2AX staining for a direct visualization of single proton hits. If single protons produce detectable foci, DNA DSBs could be used as 'biological track detectors' for protons. Ionising radiation can also damage proteins indirectly by inducing free radicals. Heat shock proteins (Hsp) help to refold or even degrade the damaged proteins. The level of the most famous heat shock protein Hsp70 is increased by ionising radiation. We investigated the expression of γH2AX and Hsp70 after cross and line patterned irradiation with counted numbers of 2.25 MeV protons on primary human skin fibroblasts. The proton induced DSBs appear more delocalised than it was expected by the ion hit accuracy. Cooling the cells before the irradiation reduces the delocalisation of DNA DSBs, which is probably caused by the reduced diffusion of DNA damaging agents. Proton irradiation seems to provoke protein damages mainly in the cytoplasm indicated by cytoplasmic Hsp70 aggregates. On the contrary, in control heat shocked cells the Hsp70 was predominantly localized in the cell nucleus. However, the irradiated area could not be recognized, all cells on the Si3N4 window showed a homogenous Hsp70 expression pattern

  4. Induction of hsp70 by the herbicide oxyfluorfen (Goal) in the Egyptian Nile fish, Oreochromis niloticus.

    Science.gov (United States)

    Hassanein, H M; Banhawy, M A; Soliman, F M; Abdel-Rehim, S A; Müller, W E; Schröder, H C

    1999-07-01

    This paper deals with the expression of the biomarker hsp70 in the liver and kidney of the freshwater fish Oreochromis niloticus following exposure to the herbicide oxyfluorfen (Goal). Fishes were exposed to three concentrations, the 96-h LC50 (3 mg/L), the 96-h (1/2)LC50 (1.5 mg/L), and the 96-h (1/4)LC50 (0.75 mg/L) of oxyfluorfen for 6, 15, and 24 days, respectively, and samples were taken at three different time periods for each concentration. The livers responded to the herbicide by an induction of the expression of both the constitutive (hsp75; Mr 75 kDa) and the inducible (hsp73; Mr 73 kDa) hsp70 proteins. In kidney, the herbicide induced a time-dependent increase in the expression of the constitutive hsp70 (hsp75) as well, but the inducible hsp70 (hsp73) required much longer incubation periods to reach maximal levels (15 and 24 days). Our results suggest that expression of hsp70 in fish is a sensitive indicator of cellular responses to herbicide exposure in the aquatic environment. PMID:10341045

  5. Changes of levels of hs-CRP, Hsp70, and MPO in patients with preeclampsia

    Institute of Scientific and Technical Information of China (English)

    Xia Liu

    2016-01-01

    Objective:To observe the changes of the levels of hs-CRP, Hsp70, and MPO in patients with preeclampsia (PE) and their clinical significance. Methods:A total of 60 pregnant women with mild and severe PE who were admitted in our hospital from January, 2013 to March, 2014 were included in the study and divided into the mild group and the severe group with 30 cases in each group. The levels of serum hs-CRP, Hsp70, and MPO were detected, and their relations with PE were analyzed. The lipid metabolism related indicators were meanwhile detected. A total of 30 healthy later pregnant women at the same stage were served as the control group for comparative analysis. Results:The levels of serum hs-CRP, Hsp70, and MPO in the mild and severe PE group were significantly higher than those in the control group (P0.05). Conclusions: The abnormal lipid metabolism in different degrees exists in the pregnant women with PE, and hs-CRP, Hsp70, and MPO are together involved in the pathogenesis of PE. Detection of the levels of hs-CRP, Hsp70, and MPO in the pregnant women with PE can accurately reflect the condition of PE.

  6. Proteomic Profiling Reveals Upregulated Protein Expression of Hsp70 in Keloids

    Directory of Open Access Journals (Sweden)

    Ju Hee Lee

    2013-01-01

    Full Text Available Background. The biochemical characteristics of keloid-derived fibroblasts differ from those of adjacent normal fibroblasts, and these differences are thought to be the cause of abnormal fibrosis. Therefore, we investigated the characteristic proteins that are differentially expressed in keloid-derived fibroblasts using proteomics tools. Objective. We attempted to investigate the novel proteins that play important roles in the pathophysiology of keloids. Methods. Proteomics analysis was performed to identify differentially expressed proteins in keloid-derived fibroblasts. Keloid-derived fibroblasts and adjacent normal fibroblasts were analyzed with 2-DAGE. We validated these proteins with immunoblot analysis, real-time RT-PCR, and immunohistochemistry. Results. Sixteen differentially expressed protein spots were identified in keloid-derived fibroblasts. Among them, heat shock protein 70 (Hsp70 was specifically upregulated in keloid-derived fibroblasts. Also, immunohistochemistry and western blot analysis revealed increased Hsp70, TGF-β, and PCNA expressions in keloids compared to normal tissue. Conclusion. Hsp70 is overexpressed in keloid fibroblasts and tissue. The overexpression of Hsp70 may be involved in the pathogenesis of keloids, and the inhibition of Hsp70 could be a new therapeutic tool for the treatment of keloids.

  7. Adult Heat Tolerance Variation in Drosophila melanogaster is Not Related to Hsp70 Expression

    DEFF Research Database (Denmark)

    Jensen, Louise Toft; Cockerell, Fiona Elizabeth; Kristensen, Torsten Nygaard; Rako, Lea; Loeschcke, Volker; McKechnie, Stephen William; Hoffmann, Ary

    Expression of heat-inducible Hsp70 is considered closely linked to thermotolerance in Drosophila melanogaster and other ectotherms. However, intra-specific variation of Hsp70 expression levels and its relationship to heat resistance has only been investigated in a few studies. Although in...... Drosophila larvae Hsp70 expression may be a key determinant of heat tolerance, the evidence for this in adults is equivocal. We therefore examined heat-induced Hsp70 expression and several measurements of adult heat tolerance in three independent collections of D. melanogaster, measured in three laboratories...... correlation between Hsp70 expression and laboratory assays of adult heat tolerance commonly used in Drosophila. When combining data across three studies we had high power to detect associations but the results showed that variation in Hsp70 expression is only likely to explain a small proportion of variation...

  8. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle

    frequently express Hsp70 on their cell surface, whereas the corresponding normal tissues do not. In addition, several clinically applied reagents, such as alkyl-lysophospholipides, chemotherapeutic agents, and anti-inflammatory reagents, have been found to enhance Hsp70 cell surface expression on cancer...... cells. We have found that inhibition of histone deacetylase (HDAC) activity leads to surface expression of Hsp70 on various hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC-inhibitor mediated Hsp70 cell surface expression was confined to...... transport and cell surface binding of Hsp70 after HDAC-inhibitor treatment remains elusive. Our data suggest that inhibition of HDAC activity selectively induces cell surface expression of Hsp70 on hematopoietic cancer cells, and this may increase the immunorecognition of these cells. It could be envisaged...

  9. Protective Effects of Overexpression TCR Vβ5.2-HSP70 and TCR Vβ8.2-HSP70 against Collagen-Induced Arthritis in Rats

    Institute of Scientific and Technical Information of China (English)

    Jing Xiao; Shentao Li; Wei Wang; Yun Li; Wenming Zhao

    2007-01-01

    Collagen-induced arthritis (CIA) is an animal model, which closely resembles human rheumatoid arthritis (RA) in pathogenesis and pathology. Evidence suggests that the inhibition of T lymphocytes or their functions can alleviate the progression of arthritis. So the administration of arthritogenic T cell receptor (TCR) variable region peptide or DNA vaccines encoding pathogenic TCR Vβ variable region may provide useful information for designing specific immunotherapies against autoimmune diseases. Heat shock proteins (HSPs) have the function of raising antigenic immunogenicity and HSP70 has a protective effect against arthritis. We previously demonstrated the presence of pathogenic predominant T cell receptor Vβ5.2 and Vβ8.2 clonotypes in the joints of CIA rats. In this study, we constructed the recombinant eukaryotic expression vectors pTARGET-TCR Vβ5.2/8.2-HSP70, and evaluated their protective effects on CIA rats. Protective effects were observed in CIA rats by injecting these recombinant DNA vaccines, which could alleviate arthritis index, decrease the levels of IFN-γ and anti-CⅡ antibody in serum, and increase the levels of IL-4. Pathological changes were not as serious as those observed in control CIA rats. The rat injected with two combined vaccines showed better protective effects than CIA rats administered with individual vaccine. These results showed that recombinant DNA vaccines pTARGET-TCR Vβ5.2-HSP70 and pTARGET-TCR Vβ8.2-HSP70 could significantly alleviate the arthritic symptoms of CIA rats, and better protective effects could be achieved if these two vaccines were used in combination.

  10. Peptide deformylase inhibitor actinonin reduces celastrol’s HSP70 induction while synergizing proliferation inhibition in tumor cells

    International Nuclear Information System (INIS)

    Celastrol is a promising anti-tumor agent, yet it also elevates heat shock proteins (HSPs), especially HSP70, this effect believed to reduce its anti-tumor effects. Concurrent use of siRNA to increase celastrol’s anti-tumor effects through HSP70 interference has been reported, but because siRNA technology is difficult to clinically apply, an alternative way to curb unwanted HSP70 elevation caused by celastrol treatment is worth exploring. In this work, we explore three alternative strategies to control HSP70 elevation: (1) Searching for cancer cell types that show no HSP70 elevation in the presence of celastrol (thus recommending themselves as suitable targets); (2) Modifying HSP70-inducing chemical groups, i.e.: the carboxyl group in celastrol; and (3) Using signaling molecule inhibitors to specifically block HSP70 elevation while protecting and/or enhancing anti-tumor effects. The first strategy was unsuccessful since celastrol treatment increased HSP70 in all 7 of the cancer cell types tested, this result related to HSF1 activation. The ubiquity of HSF1 expression in different cancer cells might explain why celastrol has no cell-type limitation for HSP70 induction. The second strategy revealed that modification of celastrol’s carboxyl group abolished its ability to elevate HSP70, but also abolished celastrol’s tumor inhibition effects. In the third strategy, 11 inhibitors for 10 signaling proteins reportedly related to celastrol action were tested, and five of these could reduce celastrol-caused HSP70 elevation. Among these, the peptide deformylase (PDF) inhibitor, actinonin, could synergize celastrol’s proliferation inhibition. Concurrent use of the chemical agent actinonin could reduce celastrol’s HSP70 elevation and also enhance proliferation inhibition by celastrol. This combination presents a novel alternative to siRNA technology and is worth further investigation for its potentially effective anti-tumor action

  11. PP2A mediated AMPK inhibition promotes HSP70 expression in heat shock response.

    Directory of Open Access Journals (Sweden)

    Ting Wang

    Full Text Available BACKGROUND: Under stress, AMP-activated protein kinase (AMPK plays a central role in energy balance, and the heat shock response is a protective mechanism for cell survival. The relationship between AMPK activity and heat shock protein (HSP expression under stress is unclear. METHODOLOGY/PRINCIPAL FINDINGS: We found that heat stress induced dephosphorylation of AMPKα subunit (AMPKα in various cell types from human and rodent. In HepG2 cells, the dephosphorylation of AMPKα under heat stress in turn caused dephosphorylation of acetyl-CoA carboxylase and upregulation of phosphoenolpyruvate carboxykinase, two downstream targets of AMPK, confirming the inhibition of AMPK activity by heat stress. Treatment of HepG2 cells with phosphatase 2A (PP2A inhibitor okadaic acid or inhibition of PP2A expression by RNA interference efficiently reversed heat stress-induced AMPKα dephosphorylation, suggesting that heat stress inhibited AMPK through activation of PP2A. Heat stress- and other HSP inducer (CdCl(2, celastrol, MG132-induced HSP70 expression could be inhibited by AICAR, an AMPK specific activator. Inhibition of AMPKα expression by RNA interference reversed the inhibitory effect of AICAR on HSP70 expression under heat stress. These results indicate that AMPK inhibition under stress contribute to HSP70 expression. Mechanistic studies showed that activation of AMPK by AICAR had no effect on heat stress-induced HSF1 nuclear translocation, phosphorylation and binding with heat response element in the promoter region of HSP70 gene, but significantly decreased HSP70 mRNA stability. CONCLUSIONS/SIGNIFICANCE: These results demonstrate that during heat shock response, PP2A mediated AMPK inhibition upregulates HSP70 expression at least partially through stabilizing its mRNA, which suggests a novel mechanism for HSP induction under stress.

  12. Characterising the KMP-11 and HSP-70 recombinant antigens' humoral immune response profile in chagasic patients

    Directory of Open Access Journals (Sweden)

    Steindel Mario

    2009-01-01

    Full Text Available Abstract Background Antigen specificity and IgG subclass could be significant in the natural history of Chagas' disease. The relationship between the different stages of human Chagas' disease and the profiles of total IgG and its subclasses were thus analysed here; they were directed against a crude T. cruzi extract and three recombinant antigens: the T. cruzi kinetoplastid membrane protein-11 (rKMP-11, an internal fragment of the T. cruzi HSP-70 protein192-433, and the entire Trypanosoma rangeli HSP-70 protein. Methods Seventeen Brazilian acute chagasic patients, 50 Colombian chronic chagasic patients (21 indeterminate and 29 cardiopathic patients and 30 healthy individuals were included. Total IgG and its subtypes directed against the above-mentioned recombinant antigens were determined by ELISA tests. Results The T. cruzi KMP-11 and T. rangeli HSP-70 recombinant proteins were able to distinguish both acute from chronic chagasic patients and infected people from healthy individuals. Specific antibodies to T. cruzi crude antigen in acute patients came from IgG3 and IgG4 subclasses whereas IgG1 and IgG3 were the prevalent isotypes in indeterminate and chronic chagasic patients. By contrast, the specific prominent antibodies in all disease stages against T. cruzi KMP-11 and T. rangeli HSP-70 recombinant antigens were the IgG1 subclass. Conclusion T. cruzi KMP-11 and the T. rangeli HSP-70 recombinant proteins may be explored together in the immunodiagnosis of Chagas' disease. Polarising the IgG1 subclass of the IgG response to T. cruzi KMP-11 and T. rangeli HSP-70 recombinant proteins could have important biological effects, taking into account that this is a complement fixing antibody.

  13. Identification and analysis of HSP70 from Sepiella maindroni under stress of Vibrio harveyi and Cd(2.).

    Science.gov (United States)

    Liu, Hui-hui; He, Jian-yu; Chi, Chang-feng; Lv, Zhen-ming

    2015-11-01

    The 70-kDa heat shock proteins (HSP70) play crucial roles in protecting cells against environmental stresses, such as heat shock, heavy metals and pathogenic bacteria. The full-length HSP70 cDNA of Sepiella maindroni (designated as SmHSP70, GenBank accession no. KJ739788) was 2109 bp, including an ORF of 1950 bp encoding a polypeptide of 649 amino acids with predicted pI/MW 5.24/71.30 kDa, a 62 bp-5'-UTR and a 97 bp-3'-UTR. BLASTp analysis and phylogenetic relationship strongly suggested that the amino acid sequence was a member of HSP70 family. Multiple sequence alignment revealed that SmHSP70 and other known HSP70 were highly conserved, especially in the regions of HSP70 family signatures, the bipartite nuclear targeting sequence, ATP/GTP-binding site motif and 'EEVD' motif. Time-dependent mRNA expression of SmHSP70 in the liver was recorded by quantitative real-time RT-PCR after Vibrio harveyi injection and Cd(2+) exposure. The results indicated that SmHSP70 played a significant role in mediating the environmental stress and immune response against pathogens. PMID:26192462

  14. Effects of HSP70 Antisense Oligonucleotide on the Proliferation and Apoptosis of Human Hepatocellular Carcinoma Cells

    Institute of Scientific and Technical Information of China (English)

    杨雪; 贺海斌; 杨威; 宋涛; 郭成; 郑鑫; 刘青光

    2010-01-01

    The study investigated the effects of heat shock protein 70(HSP70) antisense oligonucleotide(ASODN) on the proliferation and apoptosis of a human hepatocellular carcinoma cell line(SMMC-7721 cells) in vitro.HSP70 oligonucleotide was transfected into SMMC-7721 cells by the mediation of SofastTM transfection reagent.Inhibition rate of SMMC-7721 cells was determined by using MTT method.Apoptosis rate and cell cycle distribution were measured by flow cytometry.Immunocytochemistry staining was used to observe th...

  15. Luminescence diagnostics of conformational changes of the Hsp70 protein in the course of thermal denaturation

    Science.gov (United States)

    Bukina, M. N.; Bakulev, V. M.; Barmasov, A. V.; Zhakhov, A. V.; Ishchenko, A. M.

    2015-06-01

    The spectral luminescence properties of aqueous solutions of the Hsp70 protein are studied, the dependence of the luminescence spectrum on the excitation wavelength is revealed, and the temperature dependence of luminescence intensity of tyrosine and tryptophan residues in the temperature interval of 20-80° C is analyzed. The luminescence method is used to determine temperature interval (42-57° C) in which protein melting takes place. An increase in the fluorescence quantum yield of tryptophan and the bathochromic shift of the emission spectrum of denatured Hsp70 prove that transition takes place of tryptophanyls to the surface of the protein molecule.

  16. Hsp70 expression and metabolite composition in response to short-term thermal changes in Folsomia candida (Collembola)

    DEFF Research Database (Denmark)

    Waagner, Dorthe; Heckmann, Lars-Henrik; Malmendal, Anders;

    2010-01-01

    regime (8 to 32 °C; pre-treated) or a constant temperature (20 °C; control) over a period of 24 h. Exposure to a temperature increase from 20 to 32 °C (2.4 °C min-1) induced a significantly increased heat tolerance which continued throughout the experiment. Expression of the gene encoding heat shock...... protein Hsp70 was assessed at the mRNA level using real time quantitative polymerase chain reaction (QPCR). Hsp70 was rapidly induced and significantly increased by the temperature increase. The relative concentrations of low molecular weight metabolites were analysed in F. candida using nuclear magnetic...

  17. Resistência à Síndrome Ascítica, Competência Homeotérmica e Níveis de Hsp70 no Coração e Pulmão de Frangos de Corte Resistance to Ascites Syndrome, Homoeothermic Competence and Levels of Hsp70 in the Heart and Lung of Broilers

    Directory of Open Access Journals (Sweden)

    Renata Hernandes

    2002-06-01

    Full Text Available Como em outros seres vivos, também nas células das aves ocorre a síntese das proteínas de baixo peso molecular (Hsp, cujo aumento é induzido sob condições de estresse. As Hsps têm um papel importante na manutenção da integridade celular, questiona-se o seu envolvimento no mecanismo de proteção celular de órgãos alvos na ocorrência da síndrome ascítica (SA. Este trabalho objetivou avaliar a temperatura corporal e os níveis da Hsp70 no coração e pulmão de frangos de corte Hubbard (sensível à SA e caipira de pescoço-pelado (resistente, criados em termoneutralidade (25°C e frio (16°C entre 10 e 45 dias de idade. Foram utilizados 192 pintos machos, 96 de cada linhagem. Não houve mortalidade por SA nas aves caipiras. Nas aves Hubbard, a mortalidade devida à SA foi de 4% e 41% em ambiente termoneutro e frio, respectivamente. Em ambiente frio, a temperatura corporal das aves Hubbard foi menor que a das caipiras. A temperatura corporal e o nível de Hsp70 do coração das aves Hubbard diminuíram com o aumento da idade, mas não nas aves caipiras, os quais se mantiveram constantes, inclusive a Hsp70 do pulmão. Independente da idade ou da temperatura, o nível de Hsp70 no pulmão das aves caipiras era superior ao das aves Hubbard. Em relação às aves Hubbard, as caipiras são homeotérmicas mais competentes e apresentam uma maior indução de Hsp70 nos órgãos primariamente afetados na SA, mas este não parece ser o sistema de proteção contra SA, a qual as aves de pescoço pelado são resistentes.Similar to other living animals, the cells of the birds also synthesize small proteins (heat shock protein, Hsp, which increasing levels can be induce by stress. The Hsp have a relevant function in maintaining the integrity of the cell, and we question if they are involved in the mechanism of the cellular protection of target organs affected by ascites syndrome (AS. The objective of this study was to evaluate the body temperature

  18. An Hsp70 peptide initiates NK cell killing of leukemic blasts after stem cell transplantation

    NARCIS (Netherlands)

    Gross, Catharina; Holler, Ernst; Stangl, Stefan; Dickinson, Anne; Pockley, A. Graham; Asea, Alexzander A.; Mallappa, Nagaraj; Multhoff, Gabriele

    2008-01-01

    In contrast to solid tumors, leukemic blasts frequently present both Hsp70 and HLA-E on their cell Surface and thereby present activating and inhibitory signals to CD94(+) NK cells. In the first 12 months after stem cell trail splantation (SCT) CD94(+) NK cells clearly dominate over CD3(+)/CD16(-)/5

  19. Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi

    Directory of Open Access Journals (Sweden)

    Carrie E. Jolly

    2010-01-01

    Full Text Available Microsporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 70 family member (Hsp70; ECU02_0100; “C1” was chosen for further analysis. DNA encoding the C1 hsp70 was amplified, cloned and used to heterologously express the C1 Hsp70 protein, and specific antiserum was generated. Two-dimensional Western blotting analysis showed that the purified antibodies were monospecific. Immunoelectron microscopy of developing and mature E. cuniculi spores revealed that the protein localized to internal structures and not to the spore surface. In spore adherence inhibition assays, the anti-C1 antibodies did not inhibit spore adherence to host cell surfaces, whereas antibodies to a known surface adhesin (EnP1 did so. In future studies, the antibodies to the ‘C1’ Hsp70 will be used to delineate spore surface protein expression.

  20. Leishmania infantum HSP70-II null mutant as candidate vaccine against leishmaniasis: a preliminary evaluation

    Directory of Open Access Journals (Sweden)

    Fresno Manuel

    2011-07-01

    Full Text Available Abstract Background Visceral leishmaniasis is the most severe form of leishmaniasis and no effective vaccine exists. The use of live attenuated vaccines is emerging as a promising vaccination strategy. Results In this study, we tested the ability of a Leishmania infantum deletion mutant, lacking both HSP70-II alleles (ΔHSP70-II, to provide protection against Leishmania infection in the L. major-BALB/c infection model. Administration of the mutant line by either intraperitoneal, intravenous or subcutaneous route invariably leads to the production of high levels of NO and the development in mice of type 1 immune responses, as determined by analysis of anti-Leishmania IgG subclasses. In addition, we have shown that ΔHSP70-II would be a safe live vaccine as immunodeficient SCID mice, and hamsters (Mesocricetus auratus, infected with mutant parasites did not develop any sign of pathology. Conclusions The results suggest that the ΔHSP70-II mutant is a promising and safe vaccine, but further studies in more appropriate animal models (hamsters and dogs are needed to appraise whether this attenuate mutant would be useful as vaccine against visceral leishmaniasis.

  1. Congenital sideroblastic anemia due to mutations in the mitochondrial HSP70 homologue HSPA9

    DEFF Research Database (Denmark)

    Schmitz-Abe, Klaus; Ciesielski, Szymon J; Schmidt, Paul J;

    2015-01-01

    The congenital sideroblastic anemias (CSAs) are relatively uncommon diseases, characterized by defects in mitochondrial heme synthesis, iron-sulfur cluster (Fe-S) biogenesis, or protein synthesis. Here we demonstrate that mutations in HSPA9, a mitochondrial HSP70 homologue located in the 5q...

  2. Alkannin, HSP70 inducer, protects against UVB-induced apoptosis in human keratinocytes.

    Directory of Open Access Journals (Sweden)

    Yoko Yoshihisa

    Full Text Available Alkannin is an active constituent from the root extract of Alkanna tinctoria of the Boraginaceae family and it may have utility as a heat shock protein 70 (HSP70 inducer in living organisms. Here, the effects of alkannin-induced HSP70 on ultraviolet (UV B (40 mJ/cm(2-induced apoptosis were investigated in human keratinocyte HaCaT cells. Pretreatment of cells with alkannin (1 µM caused significant inhibition of UVB-induced apoptosis and caspase-3 cleavage. On the other hand, the addition of KNK437 (HSP70 inhibitor reversed the action of alkannin increasing UVB-induced apoptosis in a dose-dependent manner. In addition, differences in gene expression associated with the suppression of UVB-induced apoptosis in the presence of alkannin were investigated using Gene Chip assay. Our results indicate that alkannin suppresses UVB-induced apoptosis through the induction of HSP70 in human keratinocytes, and therefore, we suggest the usefulness of using alkannin as an antiaging agent.

  3. Hsp70 as a stress marker of sea urchin coelomocytes in short term cultures.

    Science.gov (United States)

    Matranga, Valeria; Bonaventura, Rosa; Di Bella, Gloria

    2002-06-01

    Coelomocytes are the immune effectors of the sea urchin and have shown to respond to environmental and experimental challenge by the activation of stress markers. We extended our in vivo studies to in vitro short term cultures of sea urchin coelomocytes by analysing their response to temperature being stress, acid pH and heavy metals, using the hsp70 protein as a stress marker. We found that the in vitro time course of temperature stress recapitulates results obtained in vivo where the highest overexpression was observed after 1 hour. Coelomocytes overexpress hsp70 in a time-dependent manner when cultured for 1 to 6 hr at pH 4.7 +/- 0.2 in isotonic buffer, supplemented with EDTA as anticoagulant. A peak in the level of hsp70 expression was observed at 2 hr of culture, corresponding to a 10-fold increase over the levels of control coelomocytes cultured at pH 7.3 +/- 0.2. The effect of different concentrations of CdCl2 in the culture over a period of 4 hr was also tested. We found that CdCl2 greatly increases the hsp70 expression, with 10(-3) M the dose at which the highest overexpression is observed. PMID:12064441

  4. HYPOXIA INDUCES HSP 70 PRODUCTION IN JUVENILE NILE TILAPIA, OREOCHROMIS NILOTICUS (L.)

    Science.gov (United States)

    Enzyme linked immunosorbent assay (ELISA) was used to determine levels of the inducible heat shock protein 70 (hsp70) in response to prolonged anoxic conditions in juvenile Nile tilapia. This protein was detected in brain, liver, muscle and head kidney. A significant difference between treatments (...

  5. Design, synthesis and biological evaluation of novel HSP70 inhibitors: N, N'-disubstituted thiourea derivatives.

    Science.gov (United States)

    Zeng, Yan-Qun; Cao, Rui-Yuan; Yang, Jian-Ling; Li, Xing-Zhou; Li, Song; Zhong, Wu

    2016-08-25

    As novel heat shock protein 70 (HSP70) inhibitors, N, N'-disubstituted thiourea derivatives were designed and synthesized based on the X-ray structure of the ATPase domain (nucleotide binding domain, NBD). An ATPase activity inhibition assay revealed that these compounds effectively inhibited HSP70 ATPase activity. The results revealed that the compounds 370/371/374/379/380//392/394/397/404/405 and 407 can inhibit the HSP70 ATPase turnover with high percentages of inhibition: 50.42, 38.46, 50.45, 44.12, 47.13, 50.50, 40.95, 65.36, 46.23, 35.78, and 58.37 in 200 μM, respectively. Significant synergies with lapatinib were observed for compound 379 and compound 405 in the BT474 breast cancer cell line. A structure-function analysis revealed that most of the thiourea derivatives exhibited cooperative action with lapatinib in the BT474 cancer cell line and the BT/Lap(R)1.0 lapatinib-resistant cell line. HSP70 inhibitors may be developed as synergetic drugs in drug-resistant cancer therapy. PMID:27155465

  6. Association of plasma IL-6 and Hsp70 with HRV at different levels of PAHs metabolites.

    Directory of Open Access Journals (Sweden)

    Jian Ye

    Full Text Available BACKGROUND: Exposure to polycyclic aromatic hydrocarbons (PAHs is associated with reduced heart rate variability (HRV, a strong predictor of cardiovascular diseases, but the mechanism is not well understood. OBJECTIVES: We hypothesized that PAHs might induce systemic inflammation and stress response, contributing to altered cardiac autonomic function. METHODS: HRV indices were measured using a 3-channel digital Holter monitor in 800 coke oven workers. Plasma levels of interleukin-6 (IL-6 and heat shock protein 70 (Hsp70 were determined using ELISA. Twelve urinary PAHs metabolites (OH-PAHs were measured by gas chromatography-mass spectrometry. RESULTS: We found that significant dose-dependent relationships between four urinary OH-PAHs and IL-6 (all Ptrend<0.05; and an increase in quartiles of IL-6 was significantly associated with a decrease in total power (TP and low frequency (LF (Ptrend = 0.014 and 0.006, respectively. In particular, elevated IL-6 was associated in a dose-dependent manner with decreased TP and LF in the high-PAHs metabolites groups (all Ptrend<0.05, but not in the low-PAHs metabolites groups. No significant association between Hsp70 and HRV in total population was found after multivariate adjustment. However, increased Hsp70 was significantly associated with elevated standard deviation of NN intervals (SDNN, TP and LF in the low-PAHs metabolites groups (all Ptrend<0.05. We also observed that both IL-6 and Hsp70 significantly interacted with multiple PAHs metabolites in relation to HRV. CONCLUSIONS: In coke oven workers, increased IL-6 was associated with a dose-response decreased HRV in the high-PAHs metabolites groups, whereas increase of Hsp70 can result in significant dose-related increase in HRV in the low-PAHs metabolites groups.

  7. Radioresistance of human glioma spheroids and expression of HSP70, p53 and EGFr

    International Nuclear Information System (INIS)

    Radiation therapy is routinely prescribed for high-grade malignant gliomas. However, the efficacy of this therapeutic modality is often limited by the occurrence of radioresistance, reflected as a diminished susceptibility of the irradiated cells to undergo cell death. Thus, cells have evolved an elegant system in response to ionizing radiation induced DNA damage, where p53, Hsp70 and/or EGFr may play an important role in the process. In the present study, we investigated whether the content of p53, Hsp70 and EGFr are associated to glioblastoma (GBM) cell radioresistance. Spheroids from U-87MG and MO59J cell lines as well as spheroids derived from primary culture of tumor tissue of one GBM patient (UGBM1) were irradiated (5, 10 and 20 Gy), their relative radioresistance were established and the p53, Hsp70 and EGFr contents were immunohistochemically determined. Moreover, we investigated whether EGFr-phospho-Akt and EGFr-MEK-ERK pathways can induce GBM radioresistance using inhibitors of activation of ERK (PD098059) and Akt (wortmannin). At 5 Gy irradiation UGBM1 and U-87MG spheroids showed growth inhibition whereas the MO59J spheroid was relatively radioresistant. Overall, no significant changes in p53 and Hsp70 expression were found following 5 Gy irradiation treatment in all spheroids studied. The only difference observed in Hsp70 content was the periphery distribution in MO59J spheroids. However, 5 Gy treatment induced a significant increase on the EGFr levels in MO59J spheroids. Furthermore, treatment with inhibitors of activation of ERK (PD098059) and Akt (wortmannin) leads to radiosensitization of MO59J spheroids. These results indicate that the PI3K-Akt and MEK-ERK pathways triggered by EGFr confer GBM radioresistance

  8. EFFECT OF STRESS ON THE PERCENT BODY WEIGHT CHANGE AND MRNA EXPRESSION OF IGF-1, SURVIVINE AND HSP-70 GENE IN THE HIERARCHIAL FOLLICLES OF JAPANESE QUAIL

    Directory of Open Access Journals (Sweden)

    N Shit

    2014-12-01

    Full Text Available The present study was carried out to explore the effect of stress on body weight and the mRNA expression of IGF-1, Survivine and HSP-70 gene in the hierarchial follicles of Japanese quail. A total 24 birds (10 weeks were taken and stress was induced by immobilization daily for 2hrs (between 9.00 - 11.00 AM throughout the study (10 days. Four birds were sacrificed on 1, 2, 4, 6, 8 and 10 days of the treatment. Hierarchial follicles (F1, F2 & F3 were aseptically collected to quantify the expression of IGF-1, Survivine and HSP-70 gene using real-time PCR technique. The percent body weight reduction increased and reached highest (21.30% on 10th day. The fold expression of IGF-1 gene was significantly ((P=0.05 down regulated in advance to the time of experiment. However, the fold expression of survivine gene was significantly (P=0.05 up regulated and the intensity was highest (17 fold in F-3 follicle on 4th day of experiment. No significant change in the mRNA expression of HSP-70 gene was evident in this study. From this study it may be concluded that stress brings physio-molecular change through HPA activation, which not only causes tissue regression also modifies the cellular mechanism.

  9. Organization and evolution of hsp70 clusters strikingly differ in two species of Stratiomyidae (Diptera inhabiting thermally contrasting environments

    Directory of Open Access Journals (Sweden)

    Bettencourt Brian R

    2011-03-01

    Full Text Available Abstract Background Previously, we described the heat shock response in dipteran species belonging to the family Stratiomyidae that develop in thermally and chemically contrasting habitats including highly aggressive ones. Although all species studied exhibit high constitutive levels of Hsp70 accompanied by exceptionally high thermotolerance, we also detected characteristic interspecies differences in heat shock protein (Hsp expression and survival after severe heat shock. Here, we analyzed genomic libraries from two Stratiomyidae species from thermally and chemically contrasting habitats and determined the structure and organization of their hsp70 clusters. Results Although the genomes of both species contain similar numbers of hsp70 genes, the spatial distribution of hsp70 copies differs characteristically. In a population of the eurytopic species Stratiomys singularior, which exists in thermally variable and chemically aggressive (hypersaline conditions, the hsp70 copies form a tight cluster with approximately equal intergenic distances. In contrast, in a population of the stenotopic Oxycera pardalina that dwells in a stable cold spring, we did not find hsp70 copies in tandem orientation. In this species, the distance between individual hsp70 copies in the genome is very large, if they are linked at all. In O. pardalina we detected the hsp68 gene located next to a hsp70 copy in tandem orientation. Although the hsp70 coding sequences of S. singularior are highly homogenized via conversion, the structure and general arrangement of the hsp70 clusters are highly polymorphic, including gross aberrations, various deletions in intergenic regions, and insertion of incomplete Mariner transposons in close vicinity to the 3'-UTRs. Conclusions The hsp70 gene families in S. singularior and O. pardalina evolved quite differently from one another. We demonstrated clear evidence of homogenizing gene conversion in the S. singularior hsp70 genes, which form

  10. Study of the role of Hsp70.1 in oridonin-treated hepatocarcinoma cells%Hsp70.1在冬凌草甲素处理的肝癌细胞中的作用研究

    Institute of Scientific and Technical Information of China (English)

    王辉; 叶燕; 黄树林; 禹志领

    2011-01-01

    目的 研究Hsp70.1在冬凌草甲素处理后的肝癌细胞HepG2中的表达情况及其与冬凌草甲素抗肿瘤活性的关系.方法 采用蛋白印迹法及实时荧光定量PCR技术观察Hsp70.1在冬凌草甲素处理的肝癌细胞HepG2的表达变化,并采用RNA干扰技术观察抑制Hsp70.1表达后对冬凌草甲素抗肝癌效果的影响.结果 冬凌草甲素作用12 h后,HepG2细胞的Hsp70.1蛋白及RNA表达均增加.Hsp70.1短发夹RNA(shRNA)转染抑制Hsp70.1表达后,与对照组相比,Hsp70.1 shRNA转染组、冬凌草甲素处理组或两者联合应用组的HepG2细胞的存活率均明显降低.结论 Hsp70.1可促进HepG2细胞的生存,对HepG2细胞和冬凌草甲素处理的HepG2细胞具有保护作用.%Objective To explore the expression of Hsp70. 1 in oridonin-treated hepatocarcinoma HepC2 cells and the correlation of Hsp70. 1 with the anticancer activity of oridonin. Methods By using Western blot and real-time PCR analysis,we tested the expression of Hsp70. 1 protein and mRNA in HepG2 cells treated with oridonin. After HepG2 cells were transfected with Hsp70.1 shRNA.the influence of Hsp70.1 on the anticancer activity of oridonin was investigated. Results The levels of Hsp70. 1 protein and mRNA increased in oridonin-treated HepG2 cells. Transient transfection of Hsp70. 1-specific shRNA suppressed Hsp70. 1 expression. Compared with control cells,Hsp70.1 shRNA-transfection,oridonin-treatment and combination of both-treatment groups significantly reduced cell viability. Conclusion Hsp70. 1 upregulation contributes to HepG2 cells survival,which suggests that Hsp70. 1 acts as a protective factor in both HepG2 cells and oridonin-treated HepG2 cells.

  11. Curcuma purpurascens BI. rhizome accelerates rat excisional wound healing: involvement of Hsp70/Bax proteins, antioxidant defense, and angiogenesis activity

    Directory of Open Access Journals (Sweden)

    Rouhollahi E

    2015-10-01

    Full Text Available Elham Rouhollahi,1 Soheil Zorofchian Moghadamtousi,2 Fatemeh Hajiaghaalipour,3 Maryam Zahedifard,2 Faezeh Tayeby,2 Khalijah Awang,4 Mahmood Ameen Abdulla,3 Zahurin Mohamed1 1Pharmacogenomics Laboratory, Department of Pharmacology, Faculty of Medicine, 2Institute of Biological Sciences, Faculty of Science, 3Department of Biomedical Science, Faculty of Medicine, 4Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia Purpose: Curcuma purpurascens BI. is a member of Zingiberaceae family. The purpose of this study is to investigate the wound healing properties of hexane extract of C. purpurascens rhizome (HECP against excisional wound healing in rats.Materials and methods: Twenty four rats were randomly divided into 4 groups: A negative control (blank placebo, acacia gum, B low dose of HECP, C high dose of HECP, and D positive control, with 6 rats in each group. Full-thickness incisions (approximately 2.00 cm were made on the neck area of each rat. Groups 1–4 were treated two-times a day for 20 days with blank placebo, HECP (100 mg/kg, HECP (200 mg/kg, and intrasite gel as a positive control, respectively. After 20 days, hematoxylin and eosin and Masson’s trichrome stainings were employed to investigate the histopathological alterations. Protein expressions of Bax and Hsp70 were examined in the wound tissues using immunohistochemistry analysis. In addition, levels of enzymatic antioxidants and malondialdehyde representing lipid peroxidation were measured in wound tissue homogenates.Results: Macroscopic evaluation of wounds showed conspicuous elevation in wound contraction after topical administration of HECP at both doses. Moreover, histopathological analysis revealed noteworthy reduction in the scar width correlated with the enhanced collagen content and fibroblast cells, accompanied by a reduction of inflammatory cells in the granulation tissues. At the molecular level, HECP facilitates wound-healing process

  12. Inducible Hsp70 in the Regulation of Cancer Cell Survival: Analysis of Chaperone Induction, Expression and Activity

    Directory of Open Access Journals (Sweden)

    Elisa Zorzi

    2011-10-01

    Full Text Available Understanding the mechanisms that control stress is central to realize how cells respond to environmental and physiological insults. All the more important is to reveal how tumour cells withstand their harsher growth conditions and cope with drug-induced apoptosis, since resistance to chemotherapy is the foremost complication when curing cancer. Intensive research on tumour biology over the past number of years has provided significant insights into the molecular events that occur during oncogenesis, and resistance to anti-cancer drugs has been shown to often rely on stress response and expression of inducible heat shock proteins (HSPs. However, with respect to the mechanisms guarding cancer cells against proteotoxic stresses and the modulatory effects that allow their survival, much remains to be defined. Heat shock proteins are molecules responsible for folding newly synthesized polypeptides under physiological conditions and misfolded proteins under stress, but their role in maintaining the transformed phenotype often goes beyond their conventional chaperone activity. Expression of inducible HSPs is known to correlate with limited sensitivity to apoptosis induced by diverse cytotoxic agents and dismal prognosis of several tumour types, however whether cancer cells survive because of the constitutive expression of heat shock proteins or the ability to induce them when adapting to the hostile microenvironment remains to be elucidated. Clear is that tumours appear nowadays more “addicted” to heat shock proteins than previously envisaged, and targeting HSPs represents a powerful approach and a future challenge for sensitizing tumours to therapy. This review will focus on the anti-apoptotic role of heat shock 70kDa protein (Hsp70, and how regulatory factors that control inducible Hsp70 synthesis, expression and activity may be relevant for response to stress and survival of cancer cells.

  13. Inducible Hsp70 in the Regulation of Cancer Cell Survival: Analysis of Chaperone Induction, Expression and Activity

    Energy Technology Data Exchange (ETDEWEB)

    Zorzi, Elisa [OncoHematology Clinic of Pediatrics, University-Hospital of Padova, 35100 Padova (Italy); Bonvini, Paolo, E-mail: paolo.bonvini@unipd.it [OncoHematology Clinic of Pediatrics, University-Hospital of Padova, 35100 Padova (Italy); Fondazione Città della Speranza, 36030 Monte di Malo, Vicenza (Italy)

    2011-10-21

    Understanding the mechanisms that control stress is central to realize how cells respond to environmental and physiological insults. All the more important is to reveal how tumour cells withstand their harsher growth conditions and cope with drug-induced apoptosis, since resistance to chemotherapy is the foremost complication when curing cancer. Intensive research on tumour biology over the past number of years has provided significant insights into the molecular events that occur during oncogenesis, and resistance to anti-cancer drugs has been shown to often rely on stress response and expression of inducible heat shock proteins (HSPs). However, with respect to the mechanisms guarding cancer cells against proteotoxic stresses and the modulatory effects that allow their survival, much remains to be defined. Heat shock proteins are molecules responsible for folding newly synthesized polypeptides under physiological conditions and misfolded proteins under stress, but their role in maintaining the transformed phenotype often goes beyond their conventional chaperone activity. Expression of inducible HSPs is known to correlate with limited sensitivity to apoptosis induced by diverse cytotoxic agents and dismal prognosis of several tumour types, however whether cancer cells survive because of the constitutive expression of heat shock proteins or the ability to induce them when adapting to the hostile microenvironment remains to be elucidated. Clear is that tumours appear nowadays more “addicted” to heat shock proteins than previously envisaged, and targeting HSPs represents a powerful approach and a future challenge for sensitizing tumours to therapy. This review will focus on the anti-apoptotic role of heat shock 70kDa protein (Hsp70), and how regulatory factors that control inducible Hsp70 synthesis, expression and activity may be relevant for response to stress and survival of cancer cells.

  14. Inducible Hsp70 in the Regulation of Cancer Cell Survival: Analysis of Chaperone Induction, Expression and Activity

    International Nuclear Information System (INIS)

    Understanding the mechanisms that control stress is central to realize how cells respond to environmental and physiological insults. All the more important is to reveal how tumour cells withstand their harsher growth conditions and cope with drug-induced apoptosis, since resistance to chemotherapy is the foremost complication when curing cancer. Intensive research on tumour biology over the past number of years has provided significant insights into the molecular events that occur during oncogenesis, and resistance to anti-cancer drugs has been shown to often rely on stress response and expression of inducible heat shock proteins (HSPs). However, with respect to the mechanisms guarding cancer cells against proteotoxic stresses and the modulatory effects that allow their survival, much remains to be defined. Heat shock proteins are molecules responsible for folding newly synthesized polypeptides under physiological conditions and misfolded proteins under stress, but their role in maintaining the transformed phenotype often goes beyond their conventional chaperone activity. Expression of inducible HSPs is known to correlate with limited sensitivity to apoptosis induced by diverse cytotoxic agents and dismal prognosis of several tumour types, however whether cancer cells survive because of the constitutive expression of heat shock proteins or the ability to induce them when adapting to the hostile microenvironment remains to be elucidated. Clear is that tumours appear nowadays more “addicted” to heat shock proteins than previously envisaged, and targeting HSPs represents a powerful approach and a future challenge for sensitizing tumours to therapy. This review will focus on the anti-apoptotic role of heat shock 70kDa protein (Hsp70), and how regulatory factors that control inducible Hsp70 synthesis, expression and activity may be relevant for response to stress and survival of cancer cells

  15. 旋毛虫HSP70基因的原核表达及其免疫原性%Prokaryotic expression of HSP70 gene of Trichinella spiralis and immunogenicity analysis of the expression product

    Institute of Scientific and Technical Information of China (English)

    徐佳; 禹洋; 唐颖; 俞昭旸; 李晓云; 宋铭忻

    2012-01-01

    利用RT-PCR方法扩增旋毛虫HSP70基因,将其扩增产物克隆到pEASY-Blunt Simple载体中,经限制性酶切后与pET-28a(+)载体连接,进一步转化至感受态细胞Transetta(DE3)中,用IPTG诱导表达重组旋毛虫HSP70。以纯化的重组HSP70制备多克隆抗体,并对HSP70进行免疫印迹分析及免疫组织化学染色。结果表明,重组质粒的鉴定和DNA测序均正确;SDS-PAGE和Western-blot分析结果显示,在70ku处获得一目的条带;免疫组织化学染色结果显示,该HSP70具有很好的免疫原性。证实,成功构建了重组表达质粒pET28a-HSP70,为深入研究旋毛虫HSP70的功能奠定了基础。%HSP70 gene of Trichinella spiralis was amplified by RT-PCR and then cloned into pEASY-Blunt Simple vector.The recombinant was digested and linked with pET-28a(+) vector.The recombinant plasmid was confirmed by PCR and DNA sequencing.The positive plasmid was transformed into competent Escherichia coli Transetta(DE3) and induced by IPTG for expression.The recombinant Trichinella HSP70 was purified to prepare polyclonal antibody.Trichinella HSP70 was analysed by Western-blot and stained by immunohistochemistry.The recombinant protein was confirmed to be about 70 ku in size.The positive signals were localized in the Trichinella by immunohistochemistry.The results showed that the recombinant expression plasmid pET28a-HSP70 was successfully constructed and laid the foundation for further studies on the function of HSP70.

  16. Nuclear HSP90 and HSP70 in COPD patients treated with formoterol or formoterol and corticosteroids

    Directory of Open Access Journals (Sweden)

    Holownia A

    2009-12-01

    Full Text Available Abstract Objective Heat shock proteins assist cellular protein folding and are required for the normal activity of steroid receptors. In this study we assessed nuclear HSP90 and HSP70 proteins and mRNA levels in cells isolated from induced sputum of chronic obstructive pulmonary disease patients treated for 4 weeks with formoterol (F or formoterol+budesonide (F/ICS. Methods Nuclear heat shock protein levels were assessed by Western blot and specific mRNAs were quantified in cell lysates using qRT-PCR. Results Both HSP90 and HSP70 protein levels were higher in the F/ICS-treated patients in comparison with the F-treated group (by 31%, P Conclusions It is possible that increased nuclear heat shock proteins may play a role in the attenuation of the response to glucocorticoids in COPD patients.

  17. The effect of glutamine infusion on the inflammatory response and HSP70 during human experimental endotoxaemia

    DEFF Research Database (Denmark)

    Andreasen, Anne Sofie; Pedersen-Skovsgaard, Theis; Mortensen, Ole Hartvig;

    2009-01-01

    INTRODUCTION: Glutamine supplementation has beneficial effects on morbidity and mortality in critically ill patients, possibly in part through an attenuation of the proinflammatory cytokine response and a stimulation of heat shock protein (HSP)70. We infused either alanine-glutamine or saline...... an infusion of alanine-glutamine at a rate of 0.025 g/(kg body weight x hour) or saline for 10 hours. After 2 hours, an intravenous bolus of Escherichia coli endotoxin (0.3 ng/kg) was administered. Blood samples were collected hourly for the following 8 hours. HSP70 protein content in isolated blood...... mononuclear cells (BMNCs) was measured by Western blotting. RESULTS: Plasma glutamine increased during alanine-glutamine infusion. Endotoxin reduced plasma glutamine during both trials, but plasma glutamine levels remained above baseline with alanine-glutamine supplementation. Endotoxin injection...

  18. Characterization of HSP70 and its expression in tissue: correlation with physiological and immune indices in goose (Anser cygnoides) serum.

    Science.gov (United States)

    Zhang, W W; Xiao, X; Gan, J K; Zhang, X Q; Kong, L N; Luo, Q B

    2015-01-01

    We cloned the goose heat shock protein 70 gene (HSP70), to determine its sequence variation and elucidate its mRNA expression. We designed primers to amplify the entire goose HSP70 sequence. We used 10 commercial Wuzong goslings in a heat-stress experiment. We collected tissue samples for RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR). We analyzed the variation in expression of goose HSP70 before and after heat stress. We constructed a DNA pool from six different species, for single nucleotide polymorphism (SNP) screening. We detected 18 SNPs and selected three of these SNPs for correlation analysis with biological and immune traits in 200 Wuzong geese. We showed that T+237C was significantly correlated with the serum corticosterone level, whereas T+1122C was significantly correlated with the heterophil to lymphocyte ratio. Goose HSP70 contained no introns. The results of qRT-PCR analysis revealed significant gender differences in the expression of goose HSP70 at 40°C but not at 25°C; moreover, in general, expression was significantly higher at 40°C than at 25°C. With the exception of the leg muscle and cerebellum, HSP70 expression was significantly higher in male geese than in female geese. Our results indicate that goose HSP70 plays an important role in response to severe heat stress. PMID:26505377

  19. Overexpression of MuHSP70 gene from Macrotyloma uniflorum confers multiple abiotic stress tolerance in transgenic Arabidopsis thaliana.

    Science.gov (United States)

    Masand, Shikha; Yadav, Sudesh Kumar

    2016-02-01

    A 70-KD heat shock protein (HSP70) is one of the most conserved chaperones. It is involved in de novo protein folding and prevents the aggregation of unfolded proteins under lethal environmental factors. The purpose of this study is to characterise a MuHSP70 from horsegram (Macrotyloma uniflorum) and elucidating its role in stress tolerance of plants. A MuHSP70 was cloned and characterised from a natural drought stress tolerant HPK4 variety of horsegram (M. uniflorum). For functional characterization, MuHSP70 was overexpressed in transgenic Arabidopsis. Overexpression of MuHSP70 was found to provide tolerance to the transgenic Arabidopsis against various stresses such as heat, cold, drought, salinity and oxidative stress. MuHSP70 transgenics were observed to maintain the shoot biomass, root length, relative water content, and chlorophyll content during exposure to multi-stresses relative to non-transgenic control. Transgenic lines have further shown the reduced levels of MDA, H2O2, and proteolytic activity. Together, these findings suggest that overexpression of MuHSP70 plays an important role in improving abiotic stress tolerance and could be a crucial candidate gene for exploration in crop improvement program. PMID:26694324

  20. Modulation of thermal induction of hsp70 expression by Ku autoantigen or its individual subunits.

    Science.gov (United States)

    Yang, S H; Nussenzweig, A; Li, L; Kim, D; Ouyang, H; Burgman, P; Li, G C

    1996-07-01

    Previously, we proposed a dual control mechanism for the regulation of the heat shock response in mammalian cells: a positive control mediated by the heat shock transcription factor HSF1 and a negative control mediated by the constitutive heat shock element-binding factor (CHBF). To study the physiological role of CHBF in the regulation of heat shock response, we purified CHBF to apparent homogeneity and showed it to be identical to the Ku autoantigen, a heterodimer consisting of 70-kDa (Ku-70) and 86-kDa (Ku-80) polypeptides. To study further the functional significance of Ku/CHBF in the cellular response to heat shock, we established rodent cell lines that stably and constitutively overexpressed one or both subunits of the human Ku protein, and examined the thermal induction of hsp70 and other heat shock proteins in these Ku-overexpressing ing cells. We show that expression of the human Ku-70 and Ku-80 subunits jointly or of the Ku-70 subunit alone specifically inhibits heat-induced hsp70 expression. Conversely, expression of human Ku-80 alone does not have this effect. Thermal induction of other heat shock proteins in all of the Ku-overexpressing cell lines appears not to be significantly affected, nor is the state of phosphorylation or the DNA-binding ability of HSF1 affected. These findings support a model in which hsp70 expression is controlled by a second regulatory factor in addition to the positive activation of HSF1. The Ku protein, specifically the Ku-70 subunit, is involved in the regulation of hsp70 gene expression. PMID:8668197

  1. Mutation detection in the human HSP70B′ gene by denaturing high-performance liquid chromatography

    OpenAIRE

    Hecker, Karl H.; Asea, Alexzander; Kobayashi, Kaoru; Green, Stacy; Tang, Dan; Calderwood, Stuart K

    2000-01-01

    Variances, particularly single nucleotide polymorphisms (SNP), in the genomic sequence of individuals are the primary key to understanding gene function as it relates to differences in the susceptibility to disease, environmental influences, and therapy. In this report, the HSP70B′ gene is the target sequence for mutation detection in biopsy samples from human prostate cancer patients undergoing combined hyperthermia and radiation therapy at the Dana-Farber Cancer Institute, using temperature...

  2. Polyglutamine expansion in Drosophila: thermal stress and Hsp70 as selective agents

    Indian Academy of Sciences (India)

    Brian R Bettencourt; Catherine C Hogan; Mario Nimali

    2007-04-01

    Repetitive DNA sequences that encode polyglutamine tracts are prone to expansion and cause highly deleterious phenotypes of neurodegeneration. Despite this tendency, polyglutamine tracts (``polyQs”) are conserved features of eukaryotic genomes. PolyQs are the most frequent protein-coding homotypic repeat in insect genomes, and are found predominantly in genes encoding transcription factors conserved from Drosophila through human. Although highly conserved across species, polyQ lengths vary widely within species. In D. melanogaster, polyQs in 25 genes have more alleles and higher heterozygosity than all other poly-amino acid tracts. The heat shock protein Hsp70 is a principal suppressor of polyQ expansions and may play a key role in modulating the phenotypes of the alleles that encode them. Hsp70 also promotes tolerance of natural thermal stress in Drosophila and diverse organisms, a role which may deplete the chaperone from buffering against polyQ toxicity. Thus in stressful environments, natural selection against long polyQ alleles more prone to expansion and deleterious phenotypes may be more effective. This hypothesis can be tested by measuring the phenotypic interactions between Hsp70 and polyQ transgenes in D. melanogaster undergoing natural thermal stress, an approach which integrates comparative genomics with experimental and ecological genetics.

  3. Hsp 70 in the soft shell clam (Mya arenaria): Potential as a biomarker of field contamination

    International Nuclear Information System (INIS)

    Soft shell clams (Mya arenaria) were collected from 2 contaminated and 2 control sites in Massachusetts at 4 time periods, extending over a 30 degree C temperature range. Populations at New Bedford Harbor were exposed to PCBs, PAHs, and heavy metals and populations at the Cotuit Bay Shores Beach Club were exposed to an unknown quantity and composition of petroleum hydrocarbons. Heat shock protein (hsp) 70 was measured in the gill of 20 clams per site by ELISA, using a commercially available monoclonal antibody. Expression of hsp 70 was variable between individual clams collected from the same site. Significance of the variables site and month was tested by one-way and two-way ANOVA; site, month, and site by month interaction were all significant. The significant interaction term meant that it was necessary to hold the month variable constant in order to test the hypothesis that clams from the contaminated sites contained more heat shock proteins than clams from the corresponding control sites. In general, the contaminated clams contained a higher concentration of hsp 70 than clams from the control sites, with New Bedford Harbor exhibiting the highest levels. Seasonal differences which could be responsible for the month effect include heat shock, food availability, and reproductive condition. In order to use heat shock protein expression as a biomarker of exposure to contaminants in the field, it is necessary to compare clams from contaminated sites to clams from control sites sampled at the same time

  4. Antileukemic activity of the HSP70 inhibitor pifithrin-μ in acute leukemia

    International Nuclear Information System (INIS)

    Heat shock protein (HSP) 70 is aberrantly expressed in different malignancies and has emerged as a promising new target for anticancer therapy. Here, we analyzed the in vitro antileukemic effects of pifithrin-μ (PFT-μ), an inhibitor of inducible HSP70, in acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) cell lines, as well as in primary AML blasts. PFT-μ significantly inhibited cell viability at low micromolar concentrations in all cell lines tested, with IC50 values ranging from 2.5 to 12.7 μ, and was highly active in primary AML blasts with a median IC50 of 8.9 μ (range 5.7–37.2). Importantly, higher IC50 values were seen in normal hematopoietic cells. In AML and ALL, PFT-μ induced apoptosis and cell cycle arrest in a dose-dependent fashion. PFT-μ also led to an increase of the active form of caspase-3 and reduced the intracellular concentrations of AKT and ERK1/2 in NALM-6 cells. Moreover, PFT-μ enhanced cytotoxicity of cytarabine, 17-(allylamino)-17-desmethoxygeldanamycin, suberoylanilide hydroxamic acid, and sorafenib in NALM-6, TOM-1 and KG-1a cells. This is the first study demonstrating significant antileukemic effects of the HSP70 inhibitor PFT-μ, alone and in combination with different antineoplastic drugs in both AML and ALL. Our results suggest a potential therapeutic role for PFT-μ in acute leukemias

  5. Identification of an Allosteric Pocket on Human Hsp70 Reveals a Mode of Inhibition of This Therapeutically Important Protein

    OpenAIRE

    Rodina, Anna; Patel, Pallav D.; Kang, Yanlong; Patel, Yogita; Baaklini, Imad; Wong, Michael J. H.; Taldone, Tony; Yan, Pengrong; Yang, Chenghua; Maharaj, Ronnie; Gozman, Alexander; Patel, Maulik R.; Patel, Hardik J.; Chirico, William; Erdjument-Bromage, Hediye

    2013-01-01

    Hsp70s are important cancer chaperones that act upstream of Hsp90 and exhibit independent anti-apoptotic activities. To develop chemical tools for the study of human Hsp70, we developed a homology model that unveils a previously unknown allosteric site located in the nucleotide binding domain of Hsp70. Combining structure-based design and phenotypic testing, we discovered a previously unknown inhibitor of this site, YK5. In cancer cells, this compound is a potent and selective binder of the c...

  6. Suppression of heat-induced hsp70 expression by the 70-kDa subunit of the human Ku autoantigen.

    Science.gov (United States)

    Li, G C; Yang, S H; Kim, D; Nussenzweig, A; Ouyang, H; Wei, J; Burgman, P; Li, L

    1995-05-01

    Expression of the 70-kDa polypeptide of human Ku autoantigen in rat cells is shown to suppress specifically the induction of hsp70 upon heat shock. Thermal induction of other heat shock proteins is not significantly affected, nor is the state of phosphorylation or the DNA-binding ability of the heat shock transcription factor HSF1. These findings support a model in which hsp70 gene expression is controlled by a second regulatory factor in addition to the positive activator HSF1. The Ku autoantigen, or a protein closely related to it, is likely to be involved in the regulation of hsp70 expression. PMID:7753835

  7. Histological, ultrastructural and heat shock protein 70 (HSP70) responses to heat stress in the sea cucumber Apostichopus japonicus.

    Science.gov (United States)

    Xu, Dongxue; Sun, Lina; Liu, Shilin; Zhang, Libin; Yang, Hongsheng

    2015-08-01

    The aquaculture industry for Apostichopus japonicus has suffered severe economic and resource losses due to high temperature in recent summers. There is increasing concern about the effect of high temperature on this species. Histological, ultrastructural and HSP70 responses to heat stress were investigated in the intestine of A. japonicus. Tissue degradation was observed in muscular, submucosal and mucosal layers, with significant decrease in plicae circulares of the mucosal layer. Ultrastructural damage intensified with increasing stress time, and indicators of cell apoptosis were evident after 192 h heat stress. Immunostaining showed HSP70 mainly in mucosa and serosa, with faint staining in non-stressed individuals (the control group) and denser staining under stress (the 6, 48 and 192 h groups). Western blot detection confirmed ocurrence of HSP70 in all groups and significant up-regulation under stress. The rapid and persistent response of HSP70 implies its critical role in the heat shock response of A. japonicus. PMID:25917397

  8. An inducible HSP70 gene from the midge Chironomus dilutus: Characterization and transcription profile under environmental stress

    Science.gov (United States)

    Karouna-Renier, N. K.; Rao, K.R.

    2009-01-01

    In the present study, we identified and characterized an inducible heat shock protein 70 (HSP70) from the midge Chironomus dilutus and investigated the transcriptional profile of the gene under baseline and environmentally stressful conditions. Using real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR), we observed increased expression of CD-HSP70-1 in response to both heat shock and copper stress. We also investigated the expression of this gene during midge development. All C. dilutus developmental stages expressed CD-HSP70-1 under normal conditions, although at extremely low levels. Phylogenetic analysis of the amino acid sequence demonstrated distinct clustering of this gene with inducible HSP70s from other insect species. ?? 2008 The Authors.

  9. Coagulansin-A has beneficial effects on the development of bovine embryos in vitro via HSP70 induction

    OpenAIRE

    Khan, Imran; Lee, Kyeong-Lim; Fakruzzaman, Md.; Song, Seok-Hwan; Ihsan-ul-Haq,; Mirza, Bushra; Yan, Chang Guo; Kong, Il-Keun

    2016-01-01

    Treatment with the steroidal lactone, coagulansin-A, improves bovine oocyte maturation and embryo development in vitro by inducing heat shock protein 70 (HSP70), which reduces the levels of reactive oxygen species (ROS), DNA damage and inflammation.

  10. Inhibition of cellular proliferation by the Wilms tumor suppressor WT1 requires association with the inducible chaperone Hsp70

    OpenAIRE

    Maheswaran, Shyamala; Englert, Christoph; Zheng, Gang; Lee, Sean Bong; Wong, Jenise; Harkin, D Paul; Bean, James; Ezzell, Robert; Garvin, A. Julian; McCluskey, Robert T.; DeCaprio, James A.; Haber, Daniel A.

    1998-01-01

    The Wilms tumor suppressor WT1 encodes a zinc finger transcription factor that is expressed in glomerular podocytes during a narrow window in kidney development. By immunoprecipitation and protein microsequencing analysis, we have identified a major cellular protein associated with endogenous WT1 to be the inducible chaperone Hsp70. WT1 and Hsp70 are physically associated in embryonic rat kidney cells, in primary Wilms tumor specimens and in cultured cells with inducible expression of WT1. Co...

  11. Elimination of Enhanced Thermal Resistance of Spheroid Culture Model of Prostate Carcinoma Cell Line by Inhibitors of Hsp70 Induction

    Directory of Open Access Journals (Sweden)

    Samideh Khoei

    2010-01-01

    Full Text Available AbstractObjective: The purpose of this study was to investigate the enhanced thermal resistancemechanism of the DU145 tumor spheroid cultures as compared to the prostate carcinomacell line's monolayer cultures.Materials and Methods: DU145 cells were cultured either as spheroids or monolayers.Cultures were treated with hyperthermia in a precision water bath (at 43°C for 60 minutesand/or quercetin (50 and 500 μM for monolayer and spheroid cultures respectively. Afterhyperthermic treatment, the cell viability colony forming ability, and the expression of heatshock protein 70 (Hsp70 were examined in both culture systems. Hsp70 expression wasstudied using the western blot method.Results: Our results showed that the DU145 monolayer and spheroid cell culture treatmentwith hyperthermia alone resulted in a marked survival inhibition. Furthermore, thespheroids showed a more significant resistance to hyperthermia compared to the monolayercultures (p = 0.01. They also produced more Hsp70 than the monolayer cultures.Treatment of cells with quercetin reduced the Hsp70 level in both culture systems. However,with the reduced Hsp70 levels, thermal resistance of the spheroids showed a greaterdecrease in relation to that of the monolayers.Conclusion: The results suggest that the enhanced hyperthermia resistance mechanismof the spheroid cultures compared to that of the monolayer cultures can be attributed tospheroids' Hsp70 production.

  12. B-chromosome effects on Hsp70 gene expression does not occur at transcriptional level in the grasshopper Eyprepocnemis plorans.

    Science.gov (United States)

    Navarro-Domínguez, Beatriz; Cabrero, Josefa; Camacho, Juan Pedro M; López-León, María Dolores

    2016-10-01

    As intragenomic parasites, B chromosomes can elicit stress in the host genome, thus inducing a response for host adaptation to this kind of continuous parasitism. In the grasshopper Eyprepocnemis plorans, B-chromosome presence has been previously associated with a decrease in the amount of the heat-shock protein 70 (HSP70). To investigate whether this effect is already apparent at transcriptional level, we analyze the expression levels of the Hsp70 gene in gonads and somatic tissues of males and females with and without B chromosomes from two populations, where the predominant B chromosome variants (B2 and B24) exhibit different levels of parasitism, by means of quantitative real-time PCR (qPCR) on complementary DNA (cDNA). The results revealed the absence of significant differences for Hsp70 transcripts associated with B-chromosome presence in virtually all samples. This indicates that the decrease in HSP70 protein levels, formerly reported in this species, may not be a consequence of transcriptional down-regulation of Hsp70 genes, but the result of post-transcriptional regulation. These results will help to design future studies oriented to identifying factors modulating Hsp70 expression, and will also contribute to uncover the biological role of B chromosomes in eukaryotic genomes. PMID:27334602

  13. Characterizing HSF1 Binding and Post-Translational Modifications of hsp70 Promoter in Cultured Cortical Neurons: Implications in the Heat-Shock Response.

    Directory of Open Access Journals (Sweden)

    Andrea V Gómez

    Full Text Available Causes of lower induction of Hsp70 in neurons during heat shock are still a matter of debate. To further inquire into the mechanisms regulating Hsp70 expression in neurons, we studied the activity of Heat Shock Factor 1 (HSF1 and histone posttranslational modifications (PTMs at the hsp70 promoter in rat cortical neurons. Heat shock induced a transient and efficient translocation of HSF1 to neuronal nuclei. However, no binding of HSF1 at the hsp70 promoter was detected while it bound to the hsp25 promoter in cortical neurons during heat shock. Histone PTMs analysis showed that the hsp70 promoter harbors lower levels of histone H3 and H4 acetylation in cortical neurons compared to PC12 cells under basal conditions. Transcriptomic profiling data analysis showed a predominant usage of cryptic transcriptional start sites at hsp70 gene in the rat cerebral cortex, compared with the whole brain. These data support a weaker activation of hsp70 canonical promoter. Heat shock increased H3Ac at the hsp70 promoter in PC12 cells, which correlated with increased Hsp70 expression while no modifications occurred at the hsp70 promoter in cortical neurons. Increased histone H3 acetylation by Trichostatin A led to hsp70 mRNA and protein induction in cortical neurons. In conclusion, we found that two independent mechanisms maintain a lower induction of Hsp70 in cortical neurons. First, HSF1 fails to bind specifically to the hsp70 promoter in cortical neurons during heat shock and, second, the hsp70 promoter is less accessible in neurons compared to non-neuronal cells due to histone deacetylases repression.

  14. Nuclear HSP90 and HSP70 in COPD patients treated with formoterol or formoterol and corticosteroids

    OpenAIRE

    Holownia A; Mroz RM; Kielek A; Chyczewska E; Braszko JJ

    2009-01-01

    Abstract Objective Heat shock proteins assist cellular protein folding and are required for the normal activity of steroid receptors. In this study we assessed nuclear HSP90 and HSP70 proteins and mRNA levels in cells isolated from induced sputum of chronic obstructive pulmonary disease patients treated for 4 weeks with formoterol (F) or formoterol+budesonide (F/ICS). Methods Nuclear heat shock protein levels were assessed by Western blot and specific mRNAs were quantified in cell lysates usi...

  15. Co-expression of heat shock protein (HSP) 40 and HSP70 in Pinctada martensii response to thermal, low salinity and bacterial challenges.

    Science.gov (United States)

    Li, Jun; Zhang, Yuehuan; Liu, Ying; Zhang, Yang; Xiao, Shu; Yu, Ziniu

    2016-01-01

    Heat shock protein (HSP) 40 proteins are a family of molecular chaperones that bind to HSP70 through their J-domain and regulate the function of HSP70 by stimulating its adenosine triphosphatase activity. In the present study, a HSP40 homolog named PmHSP40 was cloned from the hemocytes of pearl oyster Pinctada martensii using EST and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of PmHSP40 was 1251 bp in length, which included a 5' untranslated region (UTR) of 75 bp, an open reading frame (ORF) of a 663 bp, and a 3' UTR of 513 bp. The deduced amino acid sequence of PmHSP40 contains a J domain in the N-terminus. In response to thermal and low salinity stress challenges, the expression of PmHSP40 in hemocytes and the gill were inducible in a time-dependent manner. After bacterial challenge, PmHSP40 transcripts in hemocytes increased and peaked at 6 h post injection. In the gill, PmHSP40 expression increased, similar to expression in hemocytes; however, transcript expression of PmHSP40 was significantly up-regulated at 12 h post injection. Furthermore, the transcripts of PmHSP70 showed similar kinetics as that of PmHSP40, with highest induction during thermal, low salinity stress and bacterial challenges. Altogether these results demonstrate that PmHSP40 is an inducible protein under thermal, low salinity and bacterial challenges, suggesting its involvement in both environmental and biological stresses, and in the innate immunity of the pearl oyster. PMID:26679110

  16. Low sequence identity but high structural and functional conservation: The case of Hsp70/Hsp90 organizing protein (Hop/Sti1) of Leishmania braziliensis.

    Science.gov (United States)

    Batista, Fernanda A H; Seraphim, Thiago V; Santos, Clelton A; Gonzaga, Marisvanda R; Barbosa, Leandro R S; Ramos, Carlos H I; Borges, Júlio C

    2016-06-15

    Parasites belonging to the genus Leishmania are subjected to extensive environmental changes during their life cycle; molecular chaperones/co-chaperones act as protagonists in this scenario to maintain cellular homeostasis. Hop/Sti1 is a co-chaperone that connects the Hsp90 and Hsp70 systems, modulating their ATPase activities and affecting the fate of client proteins because it facilitates their transfer from the Hsp70 to the Hsp90 chaperone. Hop/Sti1 is one of the most prevalent co-chaperones, highlighting its importance despite the relatively low sequence identity among orthologue proteins. This multi-domain protein comprises three tetratricopeptides domains (TPR1, TPR2A and TPR2B) and two Asp/Pro-rich domains. Given the importance of Hop/Sti1 for the chaperone system and for Leishmania protozoa viability, the Leishmania braziliensis Hop (LbHop) and a truncated mutant (LbHop(TPR2AB)) were characterized. Structurally, both proteins are α-helix-rich and highly elongated monomeric proteins. Functionally, they inhibited the ATPase activity of Leishmania braziliensis Hsp90 (LbHsp90) to a similar extent, and the thermodynamic parameters of their interactions with LbHsp90 were similar, indicating that TPR2A-TPR2B forms the functional center for the LbHop interaction with LbHsp90. These results highlight the structural and functional similarity of Hop/Sti1 proteins, despite their low sequence conservation compared to the Hsp70 and Hsp90 systems, which are phylogenetic highly conserved. PMID:27103305

  17. Cytological and molecular characterization of three gametoclones of Citrus clementina

    OpenAIRE

    GERMANA, MARIA ANTONIETTA; Aleza, Pablo; Carrera, Esther; Chen, Chunxian; Chiancone, Benedetta; Costantino, Gilles; Dambier, Dominique; Deng, Xiuxin; Federici, Claire T.; Froelicher, Yann; Guo, Wenwu; Ibanez, Victoria; Juarez, Jose; Kwok, Kevin; Luro, François

    2013-01-01

    Abstract Background Three gametoclonal plants of Citrus clementina Hort. ex Tan., cv. Nules, designated ESP, FRA, and ITA (derived from three labs in Spain, France, and Italy, respectively), were selected for cytological and molecular characterization in order to elucidate genomic rearrangements provoked by haploidization. The study included comparisons of their ploidy, homozygosity, genome integrity, and gene dosage, using chromosome counting, flow c...

  18. Hsp70 Isoforms Are Essential for the Formation of Kaposi's Sarcoma-Associated Herpesvirus Replication and Transcription Compartments.

    Directory of Open Access Journals (Sweden)

    Belinda Baquero-Pérez

    2015-11-01

    Full Text Available Kaposi's sarcoma-associated herpesvirus (KSHV is an oncogenic herpesvirus associated with various AIDS-related malignancies. Like other herpesviruses, multiple processes required for KSHV lytic replication, including viral transcription, viral DNA synthesis and capsid assembly occur in virus-induced intranuclear structures, termed replication and transcription compartments (RTCs. Here we utilised a novel methodology, combining subcellular fractionation and quantitative proteomics, to identify cellular proteins which are recruited to KSHV-induced RTCs and thus play a key role in KSHV lytic replication. We show that several isoforms of the HSP70 chaperone family, Hsc70 and iHsp70, are redistributed from the cytoplasm into the nucleus coinciding with the initial formation of KSHV-induced RTCs. We demonstrate that nuclear chaperone foci are dynamic, initially forming adjacent to newly formed KSHV RTCs, however during later time points the chaperones move within KSHV RTCs and completely co-localise with actively replicating viral DNA. The functional significance of Hsp70 isoforms recruitment into KSHV RTCs was also examined using the specific Hsp70 isoform small molecule inhibitor, VER-155008. Intriguingly, results highlight an essential role of Hsp70 isoforms in the KSHV replication cycle independent of protein stability and maturation. Notably, inhibition of Hsp70 isoforms precluded KSHV RTC formation and RNA polymerase II (RNAPII relocalisation to the viral genome leading to the abolishment of global KSHV transcription and subsequent viral protein synthesis and DNA replication. These new findings have revealed novel mechanisms that regulate KSHV lytic replication and highlight the potential of HSP70 inhibitors as novel antiviral agents.

  19. HPV16E7-HSP70 Hybrid DNA Vaccine Induces E7-Specific Cytotoxic T Cells and Antitumor Immunity

    Institute of Scientific and Technical Information of China (English)

    ZHU Liqin; LI Hui; XIONG Jinhu; WANG Tongxiang; OU Xuan; WEI Yun; WU Xinxing

    2006-01-01

    Using human papillomavirus type 16 (HPV16) E7 as an antigen and Heat Shock Protein 70 as adjuvant, we constructed a DNA vaccine by linking HSP70 gene to E7C91G gene. Mice, after being immunized with E7C91G-HSP70, E7C91G/HSP70, E7C91G, and wild E7 DNA vaccines respectively, produced E7 specific CD8+ T-cell precursor frequencies oF280. 33±2.52, 144.34±4. 04, 164.34±5.13 and 82.33± 3.51 respectively within every 1 × 105 mouse splenocytes. This proves that E7C91G-HSP70 fusion vaccine can significantly enhance the E7 specific cellular immunity within the mice body(p<0.01). After being immunized with E7C91G-HSP70 fusion vaccine, tumor-bearing mice of the group being treated have significantly longer latency and survival periods, comparing with other three categories of E7 vaccines. Experiment shows that this vaccine has a significant effect on enhancing E7 positive tumor-treatment within mice body. After being immunized with E7C91G-HSP70 vaccine, there were no pathological changes found in livers, kidneys and spleens of the mice, which proves that the vaccine is quite safe. After all,E7C91G-HSP70 fusion vaccine has a much stronger tumor- treatment effect than that of wild type E7 DNA vaccine.

  20. Hsp70 - a biomarker for tumor detection and monitoring of outcome of radiation therapy in patients with squamous cell carcinoma of the head and neck

    International Nuclear Information System (INIS)

    Tumor but not normal cells frequently overexpress heat shock protein 70 (Hsp70) and present it on their cell surface (mHsp70) from where it can be actively released. Therefore, membrane (mHsp70) and soluble Hsp70 (sHsp70) were investigated as potential tumor biomarkers and for monitoring the outcome of radiation therapy. Biopsies and blood were collected from patients with squamous cell carcinoma of the head and neck (SCCHN) at different time points (before, during therapy and in the follow-up period). Hsp70 membrane expression was determined on single cell suspensions of tumor biopsies and reference tissues by flow cytometry, sHsp70 protein and antibody levels were determined in the serum of patients and healthy donors by ELISA and NK cell markers that are related to the presence of sHsp70 were analyzed in the patient’s peripheral blood lymphocytes (PBL). Tumor biopsies exhibited significantly increased mHsp70 expression levels compared to the reference tissue. Soluble Hsp70 levels were significantly higher in SCCHN patients compared to healthy human volunteers and high mHsp70 expression levels on tumor cells were associated with high sHsp70 levels in the serum of patients. Following surgery and radiotherapy sHsp70 levels in patients dropped in patients without tumor relapse in the follow-up period. In contrast to sHsp70 protein, anti-Hsp70 antibody levels remained nearly unaltered in the serum of SCCHN patients before and after therapy. Furthermore, sHsp70 protein but not anti-Hsp70 antibody levels were found to be associated with the tumor volume in SCCHN patients before start of therapy. The expression densities of the activatory NK cell markers CD56, CD94, NKG2D, NKp30, Nkp44, and NKp46 differed in patients following therapeutic intervention. A significant increase in the density of NKG2D was observed in SCCHN patients in the follow-up period after surgery and radiotherapy. We suggest sHsp70 as a potential biomarker for detecting tumors and for monitoring

  1. Cytosolic Hsp70/Hsc70 protein expression in lymphocytes exposed to low gamma-ray dose

    Energy Technology Data Exchange (ETDEWEB)

    Manzanares A, E.; Vega C, H.R.; Letechipia de Leon, C. [Unidades Academicas de Estudios Nucleares, UAZ, A.P. 336, 98000 Zacatecas (Mexico)]. E-mail: emanz@cantera.reduaz.mx; Guzman E, L.J. [Unidad Academica de Biologia Experimental, Guadalupe, Zacatecas (Mexico); Garcia T, M. [LIBRA, Centro I and D, Campus Miguel Delibes, Valladolid 47011 (Spain)

    2004-07-01

    The purpose of this study was to evaluate the effect of low gamma ray intensity upon Hsp70 expression in human Iymphocytes. The heat shock proteins (Hsp) family, are a group of proteins present in all living organism, therefore there are highly conserved and are related to adaptation and evolution. At cellular level these proteins acts as chaperones correcting denatured proteins. When a stress agent, such heavy metals, UV, heat, etc. is affecting a cell a response to this aggression is triggered only through over expression of Hsp. Several studies has been carried out in which the cellular effect are observed, mostly of these studies uses large doses, but very few studies are related with low doses. Blood of healthy volunteers was obtained and the Iymphocytes were isolated by ficoll- histopaque gradient. Experimental lots were irradiated in a {sup 137} Cs gamma-ray. Hsp70 expression was found since 0.5 c Gy, indicating a threshold to very low doses of gamma rays. (Author)

  2. The Hsp70 response of Anguillicola species to host-specific stressors.

    Science.gov (United States)

    Keppel, M; Dangel, K C; Sures, B

    2016-06-01

    The present study is based on infection experiments of two different swim bladder parasite species, Anguillicola crassus Kuwahara et al., 1974 and Anguillicola novaezelandiae Moravec and Taraschewski, 1988, which were experimentally transferred to the two eel species Anguilla anguilla Linnaeus, 1758 and Anguilla japonica Temmink and Schlegel, 1846, respectively. The host-parasite groups were selected due to their different grades of mutual adaptation. The main aim of this study was to analyze the stress responses within the parasites, which were confronted with different hosts, i.e. with different stressors related to the respective host. For this purpose, mean intensities, recovery rates, larvae output, and levels of synthesized heat shock proteins (Hsp70) were determined in nematodes of each infection group. Increased stress responses were detected in the endemic system of A. crassus parasitizing A. japonica and A. crassus in its recently acquired host A. anguilla, which seems to be associated with the immune response of the particular host species and the expenditure of energy on producing larvae. A. novaezelandiae showed overall weak activities in its unknown host species A. japonica, with the lowest recovery rate of all examined groups neither featuring elevated Hsp responses, nor a high mean intensity, nor any reproductive output. On the contrary, in A. anguilla, the parasite reached higher recovery rates, mean intensities, and reproductive output, but no increased Hsp70 levels could be detected. The four considered factors proved partially interdependent, whereas few results did not follow a clear pattern. PMID:26920569

  3. Cytosolic Hsp70/Hsc70 protein expression in lymphocytes exposed to low gamma-ray dose

    International Nuclear Information System (INIS)

    The purpose of this study was to evaluate the effect of low gamma ray intensity upon Hsp70 expression in human Iymphocytes. The heat shock proteins (Hsp) family, are a group of proteins present in all living organism, therefore there are highly conserved and are related to adaptation and evolution. At cellular level these proteins acts as chaperones correcting denatured proteins. When a stress agent, such heavy metals, UV, heat, etc. is affecting a cell a response to this aggression is triggered only through over expression of Hsp. Several studies has been carried out in which the cellular effect are observed, mostly of these studies uses large doses, but very few studies are related with low doses. Blood of healthy volunteers was obtained and the Iymphocytes were isolated by ficoll- histopaque gradient. Experimental lots were irradiated in a 137 Cs gamma-ray. Hsp70 expression was found since 0.5 c Gy, indicating a threshold to very low doses of gamma rays. (Author)

  4. Genome-wide identification of Hsp70 genes in channel catfish and their regulated expression after bacterial infection.

    Science.gov (United States)

    Song, Lin; Li, Chao; Xie, Yangjie; Liu, Shikai; Zhang, Jiaren; Yao, Jun; Jiang, Chen; Li, Yun; Liu, Zhanjiang

    2016-02-01

    Heat shock proteins 70/110 (Hsp70/110) are a family of conserved ubiquitously expressed heat shock proteins which are produced by cells in response to exposure to stressful conditions. Besides the chaperone and housekeeping functions, they are also known to be involved in immune response during infection. In this study, we identified 16 Hsp70/110 geness in channel catfish (Ictalurus punctatus) through in silico analysis using RNA-Seq and genome databases. Among them 12 members of Hsp70 (Hspa) family and 4 members of Hsp110 (Hsph) family were identified. Phylogenetic and syntenic analyses provided strong evidence in supporting the orthologies of these HSPs. In addition, we also determined the expression patterns of Hsp70/110 genes after Flavobacterium columnare and Edwardsiella ictaluri infections by meta-analyses, for the first time in channel catfish. Ten out of sixteen genes were significantly up/down-regulated after bacterial challenges. Specifically, nine genes were found significantly expressed in gill after F. columnare infection. Two genes were found significantly expressed in intestine after E. ictaluri infection. Pathogen-specific pattern and tissue-specific pattern were found in the two infections. The significantly regulated expressions of catfish Hsp70 genes after bacterial infections suggested their involvement in immune response in catfish. PMID:26693666

  5. Role of HSP70 in motoneuron survival after excitotoxic stress in a rat spinal cord injury model in vitro.

    Science.gov (United States)

    Shabbir, Ayisha; Bianchetti, Elena; Cargonja, Renato; Petrovic, Antonela; Mladinic, Miranda; Pilipović, Kristina; Nistri, Andrea

    2015-12-01

    The outcome for gait recovery from paralysis due to spinal lesion remains uncertain even when damage is limited. One critical factor is the survival of motoneurons, which are very vulnerable cells. To clarify the early pathophysiological mechanisms of spinal damage, an in vitro injury model of the rat spinal cord caused by moderate excitotoxicity was used. With this preparation we investigated whether motoneuron survival was dependent on the expression of the neuroprotective protein HSP70. In the present study excitotoxicity evoked by kainate induced delayed (24 h) loss (35%) of motoneurons, which became pyknotic with translocation of the cell death biomarker apoptosis-inducing factor (AIF) to the nucleus. This process was concomitant with suppression of locomotor network electrical activity. Surviving cells showed strong expression of HSP70 without nuclear AIF. The HSP70 inhibitor VER155008 per se induced neurotoxicity similar to that of kainate, while the HSP90 inhibitor geldanamycin did not damage spinal tissue. Electrophysiological recording following kainate or VER155008 indicated depression of motoneuron field potentials, with decreased excitability and impaired synaptic transmission. When these two drugs were applied together, more intense neurotoxicity emerged. Our data indicate that HSP70 was one important contributor to motoneuron survival and suggest that enhancing HSP70 activity is a potential future strategy for neuroprotecting these cells. PMID:26490753

  6. Geranylgeranylacetone ameliorates lung ischemia/reperfusion injury by HSP70 and thioredoxin redox system: NF-kB pathway involved.

    Science.gov (United States)

    Cao, Weijun; Li, Manhui; Li, Jianxiong; Li, Chengwei; Xu, Xin; Gu, Weiqing

    2015-06-01

    Geranylgeranylacetone (GGA) has been clinically used as an anti-ulcer drug. In the present study, we explored the protective effects of GGA on lung ischemia/reperfusion injury (IRI) and the underlying mechanism. The results demonstrated that GGA ameliorated the lung biochemical and histological alterations induced by IRI, which was reversed by HSP70 inhibition. To further explore the mechanism of GGA action, we focused on NF-kB and thioredoxin (Trx) redox system. It was shown that GGA induced the HSP70 and Trx-1 expression, NF-kB nuclear translocation and activated thioredoxin reductase (TrxR). The Trx-1 expression and TrxR activity was suppressed by HSP70 and NF-kB inhibition, while the nuclear NF-kB p65 expression was suppressed by HSP70 inhibitor. These results indicated that GGA may protect rat lung against IRI by HSP70 and Trx redox system, in which NF-kB pathway may be involved. PMID:25748490

  7. Phylogenetic relationships of organellar Hsp90 homologs reveal fundamental differences to organellar Hsp70 and Hsp60 evolution.

    Science.gov (United States)

    Emelyanov, Victor V

    2002-10-16

    In agreement with endosymbiont theory for the origin of organelles, mitochondria and chloroplasts (plastids) are universally accepted to have monophyletically arisen from within alpha-proteobacteria and cyanobacteria, respectively. Convincing particular evidence in support of this theory emerged from phylogenetic analysis of highly conserved, ubiquitous heat shock proteins (Hsps) chaperonin 60 and Hsp70. These apparently indispensable general chaperones have proven to be highly useful molecular tracers of organellar origin. Phylogenetic relationships of Hsp90--a less conserved and less widely distributed general chaperone--are reported here that are strikingly incongruent with canonical patterns of endosymbiotic ancestry. It appears that Hsp90 of chloroplasts derives from the endoplasmic reticulum-specific isoform while mitochondrial Hsp90 homologs affiliate with a eubacterial lineage other than alpha subdivision of proteobacteria. These data suggest that endosymbiont htpG genes, encoding Hsp90, have been either functionally displaced by pre-existing nuclear genes or completely lost during establishment of organelles and subsequently added to initial organellar complement. PMID:12459260

  8. 猪带绦虫HSP70-4的真核表达及其抗血清的制备%Eukaryotic expression and anti-serum preparation of HSP70-4 from Taenia solium

    Institute of Scientific and Technical Information of China (English)

    殷静; 王帅; 刘光学; 何伟; 才学鹏

    2016-01-01

    为了研究猪带绦虫(Taenia solium)热休克蛋白70-4 (TsHSP70-4)基因序列的特征及其真核表达蛋白的抗原性,参考GeneDB中猪带绦虫基因组注释信息,设计特异性引物,RT-PCR扩增TsHSP70-4ORF序列.根据毕赤酵母密码子偏好性,对TsHSP70-4基因密码子进行优化,连接载体pPIC9K,在毕赤酵母中进行诱导表达,通过Western-blot及质谱测序进行蛋白鉴定.将纯化后的TsHSP70-4表达蛋白免疫新西兰大白兔,制备抗血清.结果显示,成功克隆出大小为1 953 bp的TsHSP70-4ORF序列.在毕赤酵母中进行了高效表达,获得大小约为95 ku的重组蛋白.Western-blot检测和质谱鉴定表明,获得的重组蛋白即为TsHSP70-4.免疫新西兰大白兔,制备出效价高达1∶409 600的抗血清.该研究为猪囊尾蚴病新疫苗的研制提供了依据.

  9. Erianthus arundinaceus HSP70 (EaHSP70) Acts as a Key Regulator in the Formation of Anisotropic Interdigitation in Sugarcane (Saccharum spp. hybrid) in Response to Drought Stress.

    Science.gov (United States)

    Augustine, Sruthy Maria; Cherian, Anoop V; Syamaladevi, Divya P; Subramonian, N

    2015-12-01

    Plant growth during abiotic stress is a long sought-after trait especially in crop plants in the context of global warming and climate change. Previous studies on leaf epidermal cells have revealed that during normal growth and development, adjacent cells interdigitate anisotropically to form cell morphological patterns known as interlocking marginal lobes (IMLs), involving the cell wall-cell membrane-cortical actin continuum. IMLs are growth-associated cell morphological changes in which auxin-binding protein (ABP), Rho GTPases and actin are known to play important roles. In the present study, we investigated the formation of IMLs under drought stress and found that Erianthus arundinaceus, a drought-tolerant wild relative of sugarcane, develops such growth-related cell morphological patterns under drought stress. Using confocal microscopy, we showed an increasing trend in cortical F-actin intensity in drought-tolerant plants with increasing soil moisture stress. In order to check the role of drought tolerance-related genes in IML formation under soil moisture stress, we adopted a structural data mining strategy and identified indirect connections between the ABPs and heat shock proteins (HSPs). Initial experimental evidence for this connection comes from the high transcript levels of HSP70 observed in drought-stressed Erianthus, which developed anisotropic interdigitation, i.e. IMLs. Subsequently, by overexpressing the E. arundinaceus HSP70 gene (EaHSP70) in sugarcane (Saccharum spp. hybrid), we confirm the role of HSP70 in the formation of anisotropic interdigitation under drought stress. Taken together, our results suggest that EaHSP70 acts as a key regulator in the formation of anisotropic interdigitation in drought-tolerant plants (Erianthus and HSP70 transgenic sugarcane) under moisture stress in an actin-mediated pathway. The possible biological significance of the formation of drought-associated interlocking marginal lobes (DaIMLs) in sugarcane plants upon

  10. Evaluation of Antibody Responses Elicited by Immunization of Mice with a Pneumococcal Antigen Genetically Fused to Murine HSP70 and Murine Interleukin-4

    Institute of Scientific and Technical Information of China (English)

    Dennis O. GOR; Salamatu S. MAMBULA

    2006-01-01

    The heat shock (stress) protein HSP70 has been shown to be a potent stimulator of cellular immune responses. In order to determine whether HSP70 has the ability to stimulate antibody responses, we constructed and expressed fusion proteins consisting of murine HSP70 or murine interleukin (IL)-4 covalently linked to a pneumococcal cell wall-associated protein antigen designated PpmA. Immunization of mice with the PpmA-HSP70 fusion protein (PpmA-70) failed to elicit an increased PpmA-specific serum antibody response. In contrast, mice immunized with PpmA fused to IL-4 (PpmA-IL4), or PpmA fused to both IL-4 and HSP70 (PpmA-IL4-70) fusion proteins elicited high levels of PpmA-specific antibody responses.These data suggest that HSP70 has a limited capacity to stimulate immune responses to heterologous antigens in vivo.

  11. Inhibition of HSP70 Gene Expression by Modified Antisense and Its Effects on Embryonic Sensitivity to Heat Shock

    Institute of Scientific and Technical Information of China (English)

    TIAN Wen-ru; DU Li-yin; HE Jian-bin; LI Shou-jun

    2004-01-01

    Experiments were performed to evaluate the efficiency of inhibition of HSP70 gene expression by antisense oligonucleotides complementary to the mRNA of HSP70 and to test the effects of inhibition of HSP70 gene expression on subsequent embryonic sensitivity to heat shock. The results showed that transfection of pre-implantation embryos at 4-cell stage with 5 μM antisense oligo had no effect on in vitro blastocyst development. However, transfection with 10 to 40 μM antisense oligo had reduced in vitro blastocyst development to 15, 10% and 0; For the embryos which exposed to 40 μM As arrested at the 16-cell stage, there was no blastocyst formation within the heat shock groups. In contrast, transfection had no effect on embryonic sensitivity to heat shock, above 25% of embryos developed to blastocyst stage in control groups.

  12. [SWI/SNF Protein Complexes Participate in the Initiation and Elongation Stages of Drosophila hsp70 Gene Transcription].

    Science.gov (United States)

    Mazina, M Yu; Nikolenko, Yu V; Krasnov, A N; Vorobyeva, N E

    2016-02-01

    The participation of the SWI/SNF chromatin remodeling complex in the stimulation of the RNA polymerase II binding to gene promotors was demonstrated in all model eukaryotic organisms. It was shown eight years ago that the SWI/SNF complex influence on transcription is not limited to its role in initiation but also includes participation in elongation and alternative splicing. In the current work, we describe the subunit composition of the SWI/SNF complexes participating in initiation, preparing for the elongation and elongation of hsp70 gene transcription in Drosophila melanogaster. The data reveal the high mobility of the SWI/SNF complex composition during the hsp 70 gene transcription process. We suggest a model describing the process of sequential SWI/SNF complex formation during heat-shock induced transcription of the hsp 70 gene. PMID:27215030

  13. Sperm sorting procedure induces a redistribution of Hsp70 but not Hsp60 and Hsp90 in boar spermatozoa.

    Science.gov (United States)

    Spinaci, Marcella; Volpe, Sara; Bernardini, Chiara; de Ambrogi, Marco; Tamanini, Carlo; Seren, Eraldo; Galeati, Giovanna

    2006-01-01

    Heat shock proteins, besides their protective function against stresses, have been recently indicated as key factors for sperm fertilizing ability. Since sexing sperm by high-speed flow-cytometry subjects them to different physical, mechanical, and chemical stresses, the present study was designed to verify, by immunofluorescence and Western blot, whether the sorting procedure induces any modification in the amount and cellular distribution of heat shock proteins 60, 70, and 90 (Hsp60, Hsp70, Hsp90). Immunolocalization and Western blot quantification of both Hsp60 and Hsp90 did not reveal differences between unsorted and sorted semen. On the contrary, a redistribution of Hsp70 immunoreactivity from the equatorial subsegment toward the equator of sperm cells was recorded after sorting; this relocation suggests capacitation-like changes of sperm membrane. This modification seems to be caused mainly by incubation with Hoechst 33342, while both passage of sperm through flow cytometer and laser beam represent only minor stimuli. A further Hsp70 redistribution seems to be due to the final steps of sperm sorting, charging, and deflection of drops, and to the dilution during collection. On the other hand, staining procedure and mechanical stress seem to be the factors most injurious to sperm viability. Moreover, Hsp70 relocation was deeply influenced by the storage method. In fact, storing sexed spermatozoa, after centrifugation, in a small volume in presence of seminal plasma induced a reversion of Hsp70 redistribution, while storage in the diluted catch fluid of collection tubes caused Hsp70 relocation in most sorted spermatozoa. PMID:16870948

  14. Batroxobin Against Anoxic Damage of Rat Hippocampal Neurons in Culture: Morphological Changes and Hsp70 Expression

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@Batroxobin,the thrombin-like enzyme,is used for therapeutic defibrination. We have found that batroxobin has good therapeutic effect in ischemic reperfusion rats and clinical practices in vivo. But we have not studied the neuroprotective effect of batroxobin on anoxic hippocampal neurons in vitro. The purpose of this study was to obtain further information on the mechanism of the batroxobin-induced neuroprotection and examine the neuroprotective effect on neurons exposed to anoxia. The effect of batroxobin on anoxic damages in cultured hippocampal neurons of neonatal rats was investigated by using morphological changes and heat shock protein 70Kd (Hsp70) immunoreactive expression as indicators. The results indicate that batroxobin, besides its defibrination, may have a direct neuroprotective effect on anoxic damage of hippocampal neurons.

  15. Correlation of Hsp70 Serum Levels with Gross Tumor Volume and Composition of Lymphocyte Subpopulations in Patients with Squamous Cell and Adeno Non-Small Cell Lung Cancer

    OpenAIRE

    Gunther, Sophie; Ostheimer, Christian; Stangl, Stefan; Specht, Hanno M; Mozes, Petra; Jesinghaus, Moritz; Vordermark, Dirk; Combs, Stephanie E; Peltz, Friedhelm; Jung, Max P.; Multhoff, Gabriele

    2015-01-01

    Heat-shock protein 70 (Hsp70) is frequently found on the plasma membrane of a large number of malignant tumors including non-small cell lung cancer (NSCLC) and gets released into the blood circulation in lipid vesicles. On the one hand, a membrane (m)Hsp70-positive phenotype correlates with a high aggressiveness of the tumor; on the other hand, mHsp70 serves as a target for natural killer (NK) cells that had been pre-stimulated with Hsp70-peptide TKD plus low-dose interleukin-2 (TKD/IL-2). Fo...

  16. Inducible HSP70 Antagonizes IL-1β Cytocidal Effects through Inhibiting NF-kB Activation via Destabilizing TAK1 in HeLa Cells

    Science.gov (United States)

    Cao, Xiang; Yue, Ling; Song, JiYun; Wu, Qiuyue; Li, Na; Luo, Lan; Lan, Lei; Yin, Zhimin

    2012-01-01

    Background Despite several reports describing the HSP70-mediated cytoprotection against IL-1, the precise mechanism for this phenomenon remains to be determined. Methods/Principal Findings Here we used HeLa cells, a human epithelial carcinoma cell line, to evaluate the role of inducible HSP70 in response of IL-1β stimulation. We found that inducible HSP70 antagonized the cytotoxicity of IL-1β and improved the survival of HeLa cells. Further investigation demonstrated that increased expression level of inducible HSP70 reduced the complex of TAK1 and HSP90, and promoted the degradation of TAK1 protein via proteasome pathway. By overexpression and RNAi knockdown, we showed that inducible HSP70 modulated the NF-kB but not MAPKs signalings through influencing the stability of TAK1 protein in HeLa cells. Moreover, overexpression of HSP70 attenuated the production of iNOS upon IL-1β stimulation, validating that inducible HSP70 serves as a cytopretective factor to antagonize the cytocidal effects of IL-1β in HeLa cells. Conclusions/Significance Our observations provide evidence for a novel signaling mechanism involving HSP70, TAK1, and NF-κB in the response of IL-1β cytocidal effects. This research also provides insight into mechanisms by which HSP70 exerts its cytoprotective action upon toxic stimuli in tumor cells. PMID:23185533

  17. Potential Response to Selection of HSP70 as a Component of Innate Immunity in the Abalone Haliotis rufescens

    Science.gov (United States)

    Brokordt, Katherina B.; González, Roxana C.; Farías, William J.; Winkler, Federico M.

    2015-01-01

    Assessing components of the immune system may reflect disease resistance. In some invertebrates, heat shock proteins (HSPs) are immune effectors and have been described as potent activators of the innate immune response. Several diseases have become a threat to abalone farming worldwide; therefore, increasing disease resistance is considered to be a long-term goal for breeding programs. A trait will respond to selection only if it is determined partially by additive genetic variation. The aim of this study was to estimate the heritability (h2) and the additive genetic coefficient of variation (CVA) of HSP70 as a component of innate immunity of the abalone Haliotis rufescens, in order to assess its potential response to selection. These genetic components were estimated for the variations in the intracellular (in haemocytes) and extracellular (serum) protein levels of HSP70 in response to an immunostimulant agent in 60 full-sib families of H. rufescens. Levels of HSP70 were measured twice in the same individuals, first when they were young and again when they were pre-harvest adults, to estimate the repeatability (R), the h2 and the potential response to selection of these traits at these life stages. High HSP70 levels were observed in abalones subjected to immunostimulation in both the intracellular and extracellular haemolymph fractions. This is the first time that changes in serum levels of HSP70 have been reported in response to an immune challenge in molluscs. HSP70 levels in both fractions and at both ages showed low h2 and R, with values that were not significantly different from zero. However, HSP70 induced levels had a CVA of 13.3–16.2% in young adults and of 2.7–8.1% in pre-harvest adults. Thus, despite its low h2, HSP70 synthesis in response to an immune challenge in red abalone has the potential to evolve through selection because of its large phenotypic variation and the presence of additive genetic variance, especially in young animals. PMID

  18. HSP70 mediates survival in apoptotic cells – Boolean network prediction and experimental validation

    Directory of Open Access Journals (Sweden)

    Suhas Vasaikar

    2015-08-01

    Full Text Available Neuronal stress or injury results in the activation of proteins, which regulate the balance between survival and apoptosis. However, the complex mechanism of cell signalling involving cell death and survival, activated in response to cellular stress is not yet completely understood. To bring more clarity about these mechanisms, a Boolean network was constructed that represented the apoptotic pathway in neuronal cells. FasL and neurotrophic growth factor (NGF were considered as inputs in the absence and presence of heat shock proteins known to shift the balance towards survival by rescuing pro-apoptotic cells. The probabilities of survival, DNA repair and apoptosis as cellular fates, in the presence of either the growth factor or FasL, revealed a survival bias encoded in the network. Boolean predictions tested by measuring the mRNA expression level of caspase-3, caspase-8 and BAX in neuronal Neuro2a (N2a cell line with NGF and FasL as external input, showed positive correlation with the observed experimental results for survival and apoptotic states. It was observed that HSP70 contributed more towards rescuing cells from apoptosis in comparison to HSP27, HSP40 and HSP90. Overexpression of HSP70 in N2a transfected cells showed reversal of cellular fate from FasL-induced apoptosis to survival. Further, the pro-survival role of the proteins BCL2, IAP, cFLIP and NFκB determined by vertex perturbation analysis was experimentally validated through protein inhibition experiments using EM20-25, Embelin and Wedelolactone, which resulted in 1.27-fold, 1.26-fold and 1.46-fold increase in apoptosis of N2a cells. The existence of a one-to-one correspondence between cellular fates and attractor states shows that Boolean networks may be employed with confidence in qualitative analytical studies of biological networks.

  19. Expression and significance of MGMT and HSP70 in colorectal carcinoma%MGMT和HSP70在大肠癌中的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    吉艳荣; 薄爱华; 吉彦丽; 张彦; 牛书雷; 冯丹

    2009-01-01

    目的:探讨热休克蛋白(HSP)70和DNA修复酶O6-甲基鸟嘌呤-DNA甲基转移酶(O6-methyguanine-DNA methytransferase,MGMT)在大肠癌中的表达及临床意义.方法:采用免疫组织化学SABC法检测61例大肠癌组织中MGMT与HSP70的表达情况.结果:MGMT与HSP70在大肠癌中表达的阳性率分别为50.82%和62.30%.其中,二者都与性别及分化程度相关性不明显(P>0.05);而与年龄和浸润深度有相关性(P<0.05),且二者呈正相关(P<0.05;r =0.32).结论:MGMT与HSP70在大肠癌的发生发展中可能起重要作用,检测它们对大肠癌的防治有重要的临床指导意义.

  20. Reactive oxygen species generated by a heat shock protein (Hsp) inducing product contributes to Hsp70 production and Hsp70-mediated protective immunity in Artemia franciscana against pathogenic vibrios.

    Science.gov (United States)

    Baruah, Kartik; Norouzitallab, Parisa; Linayati, Linayati; Sorgeloos, Patrick; Bossier, Peter

    2014-10-01

    The cytoprotective role of heat shock protein (Hsp70) described in a variety of animal disease models, including vibriosis in farmed aquatic animals, suggests that new protective strategies relying upon the use of compounds that selectively turn on Hsp genes could be developed. The product Tex-OE® (hereafter referred to as Hspi), an extract from the skin of the prickly pear fruit, Opuntia ficus indica, was previously shown to trigger Hsp70 synthesis in a non-stressful situation in a variety of animals, including in a gnotobiotically (germ-free) cultured brine shrimp Artemia franciscana model system. This model system offers great potential for carrying out high-throughput, live-animal screens of compounds that have health benefit effects. By using this model system, we aimed to disclose the underlying cause behind the induction of Hsp70 by Hspi in the shrimp host, and to determine whether the product affects the shrimp in inducing resistance towards pathogenic vibrios. We provide unequivocal evidences indicating that during the pretreatment period with Hspi, there is an initial release of reactive oxygen species (hydrogen peroxide and/or superoxide anion), generated by the added product, in the rearing water and associated with the host. The reactive molecules generated are the triggering factors responsible for causing Hsp70 induction within Artemia. We have also shown that Hspi acts prophylactically at an optimum dose regimen to confer protection against pathogenic vibrios. This salutary effect was associated with upregulation of two important immune genes, prophenoloxidase and transglutaminase of the innate immune system. These findings suggest that inducers of stress protein (e.g. Hsp70) are potentially important modulator of immune responses and might be exploited to confer protection to cultured shrimp against Vibrio infection. PMID:24950414

  1. ELEVATED HSP70 STIMULATES CELL PROLIFERATION AND IS CYTOPROTECTIVE AGAINST HEAT AND VINCRISTINE TOXICITY IN MCF-7 CELLS.

    Science.gov (United States)

    Heat-shock proteins (HSPs) play important roles in regulating cell growth and protecting cells from adverse effects of heat and chemical stress. In many types of cancer, elevated HSP70 levels are associated with poor prognosis and resistance to chemotherapeutic agents. In the pre...

  2. A small molecule inhibitor for ATPase activity of Hsp70 and Hsc70 enhances the immune response to protein antigens

    Science.gov (United States)

    Baek, Kyung-Hwa; Zhang, Haiying; Lee, Bo Ryeong; Kwon, Young-Guen; Ha, Sang-Jun; Shin, Injae

    2015-12-01

    The ATPase activities of Hsp70 and Hsc70 are known to be responsible for regulation of various biological processes. However, little is known about the roles of Hsp70 and Hsc70 in modulation of immune responses to antigens. In the present study, we investigated the effect of apoptozole (Az), a small molecule inhibitor of Hsp70 and Hsc70, on immune responses to protein antigens. The results show that mice administered with both protein antigen and Az produce more antibodies than those treated with antigen alone, showing that Az enhances immune responses to administered antigens. Treatment of mice with Az elicits production of antibodies with a high IgG2c/IgG1 ratio and stimulates the release of Th1 and Th2-type cytokines, suggesting that Az activates the Th1 and Th2 immune responses. The observations made in the present study suggest that inhibition of Hsp70 and Hsc70 activities could be a novel strategy designing small molecule-based adjuvants in protein vaccines.

  3. Heat shock proteins in the retina: Focus on HSP70 and alpha crystallins in ganglion cell survival.

    Science.gov (United States)

    Piri, Natik; Kwong, Jacky M K; Gu, Lei; Caprioli, Joseph

    2016-05-01

    Heat shock proteins (HSPs) belong to a superfamily of stress proteins that are critical constituents of a complex defense mechanism that enhances cell survival under adverse environmental conditions. Cell protective roles of HSPs are related to their chaperone functions, antiapoptotic and antinecrotic effects. HSPs' anti-apoptotic and cytoprotective characteristics, their ability to protect cells from a variety of stressful stimuli, and the possibility of their pharmacological induction in cells under pathological stress make these proteins an attractive therapeutic target for various neurodegenerative diseases; these include Alzheimer's, Parkinson's, Huntington's, prion disease, and others. This review discusses the possible roles of HSPs, particularly HSP70 and small HSPs (alpha A and alpha B crystallins) in enhancing the survival of retinal ganglion cells (RGCs) in optic neuropathies such as glaucoma, which is characterized by progressive loss of vision caused by degeneration of RGCs and their axons in the optic nerve. Studies in animal models of RGC degeneration induced by ocular hypertension, optic nerve crush and axotomy show that upregulation of HSP70 expression by hyperthermia, zinc, geranyl-geranyl acetone, 17-AAG (a HSP90 inhibitor), or through transfection of retinal cells with AAV2-HSP70 effectively supports the survival of injured RGCs. RGCs survival was also stimulated by overexpression of alpha A and alpha B crystallins. These findings provide support for translating the HSP70- and alpha crystallin-based cell survival strategy into therapy to protect and rescue injured RGCs from degeneration associated with glaucomatous and other optic neuropathies. PMID:27017896

  4. hELP3 Subunit of the Elongator Complex Regulates the Transcription of HSP70 Gene in Human Cells

    Institute of Scientific and Technical Information of China (English)

    Qiuju HAN; Xiaozhe HOU; Dongmei SU; Lina PAN; Jizhou DUAN; Liguo CUI; Baiqu HUANG; Jun LU

    2007-01-01

    The human Elongator complex is remarkably similar to its yeast counterpart in several aspects.In a previous study, we analyzed the functions of the human elongation protein 3 (hELP3) subunit of the human Elongator by using an in vivo yeast complementation system. However, direct evidence for hELP3 functions in regulating gene expression in human cells was not obtained. In this study, we used hELP3 antisense oligonucleotide inhibitors to knock down hELP3 gene expression to investigate its function in human 293T cells. The results showed that specific reduction of hELP3 mRNA and protein caused a significant suppression of HSP70-2 gene expression, and this was accompanied by histone H3 hypoacetylation and decreased RNA polymerase Ⅱ density at the HSP70-2 gene. Moreover, the data also showed that hELP3 exerted the transcriptional regulatory function directly through its presence on the HSP70-2 gene. Data presented in this report provide further insight and direct evidence of the functions of hELP3 in HSP70-2 gene transcriptional elongation in human cells.

  5. Heat shock response in Chinese hamster (DHFR/sup -/) cells transfected with mouse DHFR and drosophila HSP70 genes

    International Nuclear Information System (INIS)

    In order to understand the role of the M/sub r/ = 70,000 heat shock protein (hsp70) in mammalian cells in response to heat shock, the authors transfected CHO-DHFR/sup -/ cells with the recombinant vectors pSV2-DHFR (cell lines G and M) or with pSV2-DHFR-hsp70 (cell lines H and L). Both vectors express the DHFR gene, with the latter vector containing all the regulatory, coding and termination sequences for the gene coding for hsp70 from Drosophila melanogaster. The parental cell line (CHO-DHFR/sup -/) shows the induction of several hsps (including hsp70, hsp 68 and hsp110) after a 450C heat shock. Some transfected cell lines (M and L) showed a reduced level of hsp110 and failed to show any detectable hsp68 synthesis after a 450C heat shock. A non-tranfected, methotrexate-resistant CHO cell line that expresses high DHFR activity also fails to express or shows very reduced levels of hsp68 after a heat shock. The authors also tranfected cells (G and H) that do express hsp68 and hsp110 after a heat shock. They are currently investigating the effects of hsp68 and DHFR expression on growth and thermotolerance in these cell lines. The authors conclude that there are sequences present in the vector pSV2-DHFR (possibly the DHFR gene) that may be involved in hsp68 expression

  6. Molecular Interactions between the Citrus Bacterial Pathogen Candidatus Liberbacter asiaticus and Its Insect Vector the Asian Citrus Psyllid Diaphorina citri

    OpenAIRE

    Wang, Linling; Killiny, Nabil

    2014-01-01

    Huanglongbing (HLB), the most serious disease of citrus, is attributed in the United States to Candidatus Liberibacter asiaticus (CLas), a gram-negative, phloem-restricted α-proteobacterium transmitted by the Asian citrus psyllid (Diaphorina citri). Despite the fact that the psyllid is well recognized as the vector of CLas, to the best of our knowledge, little research has so far been conducted on molecular interactions between CLas and the psyllid. Many gram-negative bacterial pathogens have...

  7. Immunogenicity of HSP-70, KMP-11 and PFR-2 leishmanial antigens in the experimental model of canine visceral leishmaniasis.

    Science.gov (United States)

    Carrillo, Eugenia; Crusat, Martín; Nieto, Javier; Chicharro, Carmen; Thomas, Maria del Carmen; Martínez, Enrique; Valladares, Basilio; Cañavate, Carmen; Requena, Jose María; López, Manuel Carlos; Alvar, Jorge; Moreno, Javier

    2008-03-28

    Zoonotic visceral leishmaniasis (ZVL) is a parasitic disease caused by Leishmania infantum/L. chagasi that is emerging as an important medical and veterinary problem. Dogs are the domestic reservoir for this parasite and, therefore, the main target for controlling the transmission to humans. In the present work, we have evaluated the immunogenicity of the Leishmania infantum heat shock protein (HSP)-70, paraflagellar rod protein (PFR)-2 and kinetoplastida membrane protein (KMP)-11 recombinant proteins in dogs experimentally infected with the parasite. We have shown that peripheral blood mononuclear cells (PBMC) from experimentally infected dogs proliferated in response to these recombinant antigens and against the soluble leishmanial antigen (SLA). We have also quantified the mRNA expression level of the cytokines induced in PBMC upon stimulation with the HSP-70, PFR-2 and KMP-11 proteins. These recombinant proteins induced an up-regulation of IFN-gamma. HSP-70 and PFR-2 also produced an increase of the TNF-alpha transcripts abundance. No measurable induction of IL-10 was observed and low levels of IL-4 mRNA were produced in response to the three mentioned recombinant antigens. Serum levels of specific antibodies against HSP-70, PFR-2 and KMP-11 recombinant proteins were also determined in these animals. Our study showed that HSP-70, KMP-11 and PFR-2 proteins are recognized by infected canines. Furthermore, these antigens produce a Th1-type immune response, suggesting that they may be involved in protection. The identification as vaccine candidates of Leishmania antigens that elicit appropriate immune responses in the canine model is a key step in the rational approach to generate a vaccine for canine visceral leishmaniasis. PMID:18321614

  8. An inducible heat shock protein 70 small molecule inhibitor demonstrates anti-dengue virus activity, validating Hsp70 as a host antiviral target.

    Science.gov (United States)

    Howe, Matthew K; Speer, Brittany L; Hughes, Philip F; Loiselle, David R; Vasudevan, Subhash; Haystead, Timothy A J

    2016-06-01

    An estimated three billion people are at risk of Dengue virus (DENV) infection worldwide and there are currently no approved therapeutic interventions for DENV infection. Due to the relatively small size of the DENV genome, DENV is reliant on host factors throughout the viral life cycle. The inducible form of Heat Shock Protein 70 (Hsp70i) has been implicated as a host factor in DENV pathogenesis, however the complete role remains to be elucidated. Here we further illustrate the importance of Hsp70i in dengue virus pathogenesis and describe the antiviral activity of the allosteric small molecule inhibitor that is selective for Hsp70i, called HS-72. In monocytes, Hsp70i is expressed at low levels preceding DENV infection, but Hsp70i expression is induced upon DENV infection. Targeting Hsp70i with HS-72, results in a dose dependent reduction in DENV infected monocytes, while cell viability was maintained. HS-72 works to reduce DENV infection by inhibiting the entry stage of the viral life cycle, through disrupting the association of Hsp70i with the DENV receptor complex. This work highlights Hsp70i as an antiviral target and HS-72 as a potential anti-DENV therapeutic agent. PMID:27058774

  9. ANTI-HSP60 and ANTI-HSP70 antibody levels and micro/ macrovascular complications in type 1 diabetes: the EURODIAB Study

    DEFF Research Database (Denmark)

    Gruden, G.; Bruno, G.; Chaturvedi, N.;

    2009-01-01

    OBJECTIVES: The heat shock proteins 60 and 70 (HSP60, HSP70) play an important role in cytoprotection. Under stress conditions they are released into the circulation and elicit an immune response. Anti-HSP60 and anti-HSP70 antibody levels have been associated with cardiovascular disease. Type 1 d...

  10. Fluoro-edenite Fibers Induce Expression of Hsp70 and Inflammatory Response

    Directory of Open Access Journals (Sweden)

    Michael Balazy

    2007-09-01

    Full Text Available Many asbestos-like mineral fibers have been detected in the air of mountainous and volcanic areas of Italy and other parts of the world. These fibers have been suspected to be the cause of increased incidences of lung cancer and other lung diseases in these areas. However, the mechanisms of the cellular response and defense following exposure to these microscopic fibers have not been characterized. We continue to study these mechanisms to be able to propose preventive strategies in large populations. The objective of the present study was to determine comparatively biological responses of mesothelial Met-5A and monocyte-macrophage J774 cells following exposure to two types of fluoro-edenite fibers having low and high iron content (labeled 19 and 27, respectively obtained from Biancavilla (Sicily, Italy. The reference fiber was a non-iron fibrous tremolite from Val di Susa (Piemonte, Italy. The cells were treated with 5, 50, and 100 μg of fibrous matter per 1 ml for 72 hr. We identified several key mechanisms by which cells responded and counteracted the injury induced by these fibers. The fibers caused induction of the heat shock protein 70 (Hsp70, stimulated formation of reactive oxygen species (detected by using DCFH-DA as a fluorescent probe and NO• (measured as nitrite. Exposure of cells to the fibers induced lactate dehydrogenase activity and decreased viability. The fluoro-endenite type 27 was the most potent fiber tested, which indicated that iron and possibly manganese contribute significantly to this fiber toxicity. The J774 cells were more sensitive to fluoro-edenite than Met-5A cells suggesting that the primary site of the fiberinduced inflammatory response could be the macrophage rather than the pulmonary epithelium. Fluoro-edenite produces more biological alterations with respect to non-iron tremolite. Hsp70 and free radicals could be important factors in the context of mineral fiber-induced acute lung injury

  11. Differential role for ERK2 in anoxia-induced activation of transcription and translation of Hsp70 in NIH 3T3 cells

    DEFF Research Database (Denmark)

    Ossum, Carlo G; Lauritsen, Anders N; Karottki, Dorina Gabriela; Hoffmann, Else K

    2011-01-01

    transcription and translation of Hsp70 during recovery from chemical anoxia and the role of the extracellular signal regulated kinase ERK2 in this induction of Hsp70. 10 mM azide for 30 minutes (chemical anoxia) significantly inhibited the activity of ERK2 (measured as phospho-ERK) but the ERK-2 activity is...... rapidly increased in a MEK-independen manner, when azide is washed out of the cells. Chemical anoxia and overnight recovery induced Hsp70 expression (analyzed by Western blotting) and this was inhibited by actinomycin D as well as by cycloheximide showing that induction of both translation and...... transcription was involved. Inhibition of the MAP kinase p38, which was transiently activated during chemical anoxia, had no effect on the increase in Hsp70 expression whereas an inhibitor of reactive oxygen species and inhibition of the phosphatase PP1 and PP2a inhibited the increase in Hsp70 expression...

  12. Hsp70-Hsp40 chaperone complex functions in controlling polarized growth by repressing Hsf1-driven heat stress-associated transcription.

    Directory of Open Access Journals (Sweden)

    Aleksandar Vjestica

    Full Text Available How the molecular mechanisms of stress response are integrated at the cellular level remains obscure. Here we show that the cellular polarity machinery in the fission yeast Schizosaccharomyces pombe undergoes dynamic adaptation to thermal stress resulting in a period of decreased Cdc42 activity and altered, monopolar growth. Cells where the heat stress-associated transcription was genetically upregulated exhibit similar growth patterning in the absence of temperature insults. We identify the Ssa2-Mas5/Hsp70-Hsp40 chaperone complex as repressor of the heat shock transcription factor Hsf1. Cells lacking this chaperone activity constitutively activate the heat-stress-associated transcriptional program. Interestingly, they also exhibit intermittent monopolar growth within a physiological temperature range and are unable to adapt to heat stress. We propose that by negatively regulating the heat stress-associated transcription, the Ssa2-Mas5 chaperone system could optimize cellular growth under different temperature regiments.

  13. Adaptive capability as indicated by behavioral and physiological responses, plasma HSP70 level, and PBMC HSP70 mRNA expression in Osmanabadi goats subjected to combined (heat and nutritional) stressors

    Science.gov (United States)

    Shilja, Shaji; Sejian, V.; Bagath, M.; Mech, A.; David, C. G.; Kurien, E. K.; Varma, Girish; Bhatta, Raghavendra

    2015-12-01

    A study was conducted to assess the impact of heat and nutritional stress simultaneously on the adaptive capability as indicated by behavioral and physiological responses, plasma heat shock protein 70 (HSP70) level, and peripheral blood mononuclear cells (PBMC) HSP70 gene expression in goats. Twenty-four adult Osmanabadi bucks (average body weight (BW) 16.0 kg) were used in the present study. The bucks were divided into four groups viz., C (n = 6; control), HS (n = 6; heat stress), NS (n = 6; nutritional stress), and CS (n = 6; combined stress). The study was conducted for a period of 45 days. C and HS bucks had ad libitum access to their feed while NS and CS bucks were under restricted feed (30 % intake of C bucks) to induce nutritional stress. The HS and CS bucks were exposed to solar radiation for 6 h a day between 10:00 a.m. and 4:00 p.m. to induce heat stress. The data was analyzed using repeated measures analysis of variance. The standing time differed significantly (P groups (C and HS) and restricted feeding groups (NS and CS). The highest (P group while the lowest in the C and HS groups. The highest (P group. Water intake recorded was significantly (P groups. The highest respiration rate (RR), pulse rate (PR), and rectal temperature (RT) during the afternoon were also recorded in the CS group. Further, skin temperature of the head, flank, and scrotum during the afternoon was also higher (P group. In addition, both plasma HSP70 concentration and PBMC HSP70 messenger RNA (mRNA) transcript expression were also significantly (P group. It can be concluded from this study that when two stressors occur simultaneously, they may have severe impact on adaptive capabilities of Osmanabadi bucks as compared to that would occur individually. Further, the study indicated that lying time, drinking frequency, RR, RT, plasma HSP70, and PBMC HSP70 gene expression may act as ideal biological markers for assessing the impact of CS on adaptive capabilities in bucks.

  14. The Chaperone Balance Hypothesis: The Importance of the Extracellular to Intracellular HSP70 Ratio to Inflammation-Driven Type 2 Diabetes, the Effect of Exercise, and the Implications for Clinical Management

    Directory of Open Access Journals (Sweden)

    Mauricio Krause

    2015-01-01

    Full Text Available Recent evidence shows divergence between the concentrations of extracellular 70 kDa heat shock protein [eHSP70] and its intracellular concentrations [iHSP70] in people with type 2 diabetes (T2DM. A vital aspect regarding HSP70 physiology is its versatility to induce antagonistic actions, depending on the location of the protein. For example, iHSP70 exerts a powerful anti-inflammatory effect, while eHSP70 activates proinflammatory pathways. Increased eHSP70 is associated with inflammatory and oxidative stress conditions, whereas decreased iHSP70 levels are related to insulin resistance in skeletal muscle. Serum eHSP70 concentrations are positively correlated with markers of inflammation, such as C-reactive protein, monocyte count, and TNF-α, while strategies to enhance iHSP70 (e.g., heat treatment, chemical HSP70 inducers or coinducers, and physical exercise are capable of reducing the inflammatory profile and the insulin resistance state. Here, we present recent findings suggesting that imbalances in the HSP70 status, described by the [eHSP70]/[iHSP70] ratio, may be determinant to trigger a chronic proinflammatory state that leads to insulin resistance and T2DM development. This led us to hypothesize that changes in this ratio value could be used as a biomarker for the management of the inflammatory response in insulin resistance and diabetes.

  15. The p53/HSP70 inhibitor, 2-phenylethynesulfonamide, causes oxidative stress, unfolded protein response and apoptosis in rainbow trout cells

    Energy Technology Data Exchange (ETDEWEB)

    Zeng, Fanxing; Tee, Catherine; Liu, Michelle [Department of Biology, University of Waterloo, Waterloo, Ontario N2L 3G1 (Canada); Sherry, James P. [Aquatic Contaminants Research Division, Environment Canada, Burlington, Ontario L7R 4A6 (Canada); Dixon, Brian; Duncker, Bernard P. [Department of Biology, University of Waterloo, Waterloo, Ontario N2L 3G1 (Canada); Bols, Niels C., E-mail: ncbols@uwaterloo.ca [Department of Biology, University of Waterloo, Waterloo, Ontario N2L 3G1 (Canada)

    2014-01-15

    Highlights: •2-Phenylethynesulfonamide (PES) is an inhibitor of p53 and HSP 70 in mammals. •In the fish epithelial cell line, RTgill-W1, PES enhanced ROS generation and was cytotoxic. •RTgill-W1 death was by apoptosis and blocked by the anti-oxidant N-acetylcysteine. •This is the first report linking PES-induced cell death to ROS. •With this background PES should be useful for studying fish cell survival pathways. -- Abstract: The effect of 2-phenylethynesulfonamide (PES), which is a p53 and HSP70 inhibitor in mammalian cells, was studied on the rainbow trout (Oncorhynchus mykiss) gill epithelial cell line, RTgill-W1, in order to evaluate PES as a tool for understanding the cellular survival pathways operating in fish. As judged by three viability assays, fish cells were killed by 24 h exposures to PES, but cell death was blocked by the anti-oxidant N-acetylcysteine (NAC). Cell death had several hallmarks of apoptosis: DNA laddering, nuclear fragmentation, Annexin V staining, mitochondrial membrane potential decline, and caspases activation. Reactive oxygen species (ROS) production peaked in several hours after the addition of PES and before cell death. HSP70 and BiP levels were higher in cultures treated with PES for 24 h, but this was blocked by NAC. As well, PES treatment caused HSP70, BiP and p53 to accumulate in the detergent-insoluble fraction, and this too was prevented by NAC. Of several possible scenarios to explain the results, the following one is the simplest. PES enhances the generation of ROS, possibly by inhibiting the anti-oxidant actions of p53 and HSP70. ER stress arises from the ROS and from PES inhibiting the chaperone activities of HSP70. The ER stress in turn initiates the unfolded protein response (UPR), but this fails to restore ER homeostasis so proteins aggregate and cells die. Despite these multiple actions, PES should be useful for studying fish cellular survival pathways.

  16. HSP70在鼠背轴型皮管IPC后的保护作用的实验研究%Study of HSP70 protective effect on ischemic preconditioning axial flap in rats

    Institute of Scientific and Technical Information of China (English)

    陈劲松; 夏双印; 杨大平

    2001-01-01

    Objective This study was to reveal the continuous changes of HSP70 and its regularity in the delayed protection of ischemic preconditioning axial flap in rats,then speculate the best time- piont of protection. Methods Animals were divided into three groups randomly(control、amputate after isehemia pretreatment and amputate delay after ischemia pretreatment). A left dorsal skin flap was crated in Wistar rat to ischemia precondition model to examine the survival rate of the skin flaps, electronic microscopic appearanee, the contents of HSP70 at various times after they were given different treatment. Results The survival rate of amputate after ischamia pretreatment and amPutate delay 48 h after ischemia pretreatment are 1.26 and 1.57 times as high as that of control. The celluar biosynthesis at 48 h after IPC is vigorous when there is a lot of proteins which were synthesized and stored. The contents of HSP70 at 48 h after IPC or at 72 h after IPC is much more than that of control, especially at the 48 h. Conclusions IPC can improve the survival rate of ischemia flap and amputate delay after IPC has more effect than IPC does. 48 h may be the best time - point of IPC protection. It is suggest that the mechanism of IPC protection can be related to the function of HSP70.%目的揭示IPC后皮瓣中延迟相HSP70表达的动态变化,提出其规律及保护作用的最佳时间。方法采用大鼠背部轴型皮管,制成活体原位IPC模型,检测皮管存活率,组织细胞超微结构,HSP70测定及统计学分析。结果 IPC组和IPC后延迟48h断蒂组与对照组相比分别提高了皮瓣存活面积1.26倍和1.57倍;IPC后48h组细胞合成功能旺盛,有大量的蛋白质合成并贮存;IPC后48、72 h组HSP70均较对照组升高,并且其表达以48h较为明显。结论 IPC对鼠背轴型皮管具有保护作用,而对延迟断蒂组保护作用更佳,以48h为最佳时间点。HSP70可能发挥着重要的作用。

  17. Correlation of Hsp70 serum levels with gross tumor volume and composition of lymphocyte subpopulations in patients with squamous cell and adeno non-small cell lung cancer

    Directory of Open Access Journals (Sweden)

    Sophie eGunther

    2015-11-01

    Full Text Available Heat shock protein 70 (Hsp70 is frequently found on the plasma membrane of a large number of malignant tumors including non-small cell lung cancer (NSCLC and gets released into the blood circulation in lipid vesicles. On the one hand a membrane (mHsp70-positive phenotype correlates with a high aggressiveness of the tumor, on the other hand, mHsp70 serves as a target for natural killer (NK cells that had been pre-stimulated with Hsp70-peptide TKD plus low dose interleukin-2 (TKD/IL-2. Following activation, NK cells show an up-regulated expression of activatory C-type lectin receptors, such as CD94/NKG2C, NKG2D and natural cytotoxicity receptors (NCRs; NKp44, NKp46, NKp30 and thereby gain the capacity to kill mHsp70-positive tumor cells. With respect to these results, the efficacy of ex vivo TKD/IL-2 stimulated, autologous NK cells is currently tested in a proof-of-concept phase II clinical trial in patients with squamous cell NSCLC after RCT at the TUM. Inclusion criteria are histological proven, non-resectable NSCLC in stage IIIA/IIIB, clinical responses to RCT and a mHsp70-positive tumor phenotype. The mHsp70 status is determined in the serum of patients using the lipHsp70 ELISA test which enables the quantification of liposomal and free Hsp70. Squamous cell and adeno NSCLC patients had significantly higher serum Hsp70 levels than healthy controls. A significant correlation of serum Hsp70 levels with the gross tumor volume (GTV was shown for adeno and squamous cell NSCLC. However, significantly elevated ratios of activated CD69+/CD94+ NK cells that are associated with low serum Hsp70 levels were observed only in patients with squamous cell lung cancer. These data might provide a first hint that the innate immune system might get activated by soluble Hsp70 in squamous cell but not adeno NSCLC.

  18. Morphological, molecular and virulence characterization of three Lencanicillium species infecting Asian citrus psyllids in Huangyan citrus groves.

    Science.gov (United States)

    Lu, Lianming; Cheng, Baoping; Du, Danchao; Hu, Xiurong; Peng, Aitian; Pu, Zhanxu; Zhang, Xiaoya; Huang, Zhendong; Chen, Guoqing

    2015-02-01

    Citrus greening or Huanglongbing (HLB) is caused by the infection of Candidatus Liberibacter spp. in citrus plants. Since Asian citrus psyllid is the primary vector of this bacterial pathogen, the spread of HLB can be mitigated by suppressing Asian citrus psyllid populations in citrus groves using entomopathogens. To expand the current data on entomopathogens infecting Asian citrus psyllids, we isolated and characterized three different entomopathogens. Strains ZJLSP07, ZJLA08, and ZJLP09 infected the Asian citrus psyllid, Diaphorina citri Kuwayama, in Huangyan citrus groves. Based on molecular and morphological analyses, two were identified as Lecanicillium attenuatum and Lecanicillium psalliotae, and the third was recognized as an unidentified species of the genus, Lecanicillium. The corrected mortalities caused by strains ZJLSP07, ZJLA08 were 100% at 7days post-inoculation, while by ZJLP09 complete mortality occurred at 6days after inoculation, with 1.0×10(8)conidia/ml at 25°C and a relative humidity of 90% in the laboratory. Under the same condition, the corrected mortalities caused by strains ZJLSP07, ZJLA08 and ZJLP09 were 100%, 92.55% and 100%, respectively at 9days post-inoculation in the greenhouse. Our findings also revealed that these fungal strains infected D. citri using hyphae that penetrated deep into the insect tissues. Further, all three strains secreted the enzymes proteinases, chitinases and lipases with a potential to destroy insect tissues. Interestingly, strain ZJLP09 had an earlier invasion time and the highest levels of enzyme activities when compared to the other two strains. These findings have expanded the existing pool of entomopathogenic fungi that infect D. citri and can be potentially used for the management of D. citri populations. PMID:25593036

  19. Reduced heat shock response in human mononuclear cells during aging and its association with polymorphisms in HSP70 genes

    DEFF Research Database (Denmark)

    Singh, Ripudaman; Kølvraa, Steen; Bross, Peter;

    2006-01-01

    Age-dependent changes in heat shock response (HSR) were studied in mononuclear cells (monocytes and lymphocytes) collected from young (mean age = 22.6 +/- 1.7 years) and middle-aged (mean age = 56.3 +/- 4.7 years) subjects after 1 hour of heat shock at 42 degrees C. Genotype-specific HSR was...... measured by genotyping the subjects for 3 single nucleotide polymorphisms, HSPA1A(A-110C), HSPA1B(A1267G), and HSPA1L(T2437C), 1 each in the 3 HSP70 genes. A significant age-related decrease in the induction of Hsp70 occurred after heat shock in both monocytes and lymphocytes. The noninducible and...

  20. Modelling the role of the Hsp70/Hsp90 system in the maintenance of protein homeostasis.

    Directory of Open Access Journals (Sweden)

    Carole J Proctor

    Full Text Available Neurodegeneration is an age-related disorder which is characterised by the accumulation of aggregated protein and neuronal cell death. There are many different neurodegenerative diseases which are classified according to the specific proteins involved and the regions of the brain which are affected. Despite individual differences, there are common mechanisms at the sub-cellular level leading to loss of protein homeostasis. The two central systems in protein homeostasis are the chaperone system, which promotes correct protein folding, and the cellular proteolytic system, which degrades misfolded or damaged proteins. Since these systems and their interactions are very complex, we use mathematical modelling to aid understanding of the processes involved. The model developed in this study focuses on the role of Hsp70 (IPR00103 and Hsp90 (IPR001404 chaperones in preventing both protein aggregation and cell death. Simulations were performed under three different conditions: no stress; transient stress due to an increase in reactive oxygen species; and high stress due to sustained increases in reactive oxygen species. The model predicts that protein homeostasis can be maintained during short periods of stress. However, under long periods of stress, the chaperone system becomes overwhelmed and the probability of cell death pathways being activated increases. Simulations were also run in which cell death mediated by the JNK (P45983 and p38 (Q16539 pathways was inhibited. The model predicts that inhibiting either or both of these pathways may delay cell death but does not stop the aggregation process and that eventually cells die due to aggregated protein inhibiting proteasomal function. This problem can be overcome if the sequestration of aggregated protein into inclusion bodies is enhanced. This model predicts responses to reactive oxygen species-mediated stress that are consistent with currently available experimental data. The model can be used to

  1. Association of HSP70 and genotoxic damage in lymphocytes of workers exposed to coke-oven emission

    OpenAIRE

    Xiao, Chengfeng; Chen, Sheng; Li, Jizhao; Hai, Tao; Lu, Qiaofa; Sun, Enling; Wang, Ruibo; Tanguay, Robert M.; Wu, Tangchun

    2002-01-01

    Heat shock proteins (Hsps) have been reported to protect cells, tissues, and organisms against damage from a wide variety of stressful stimuli. Whether they protect against deoxyribonucleic acid (DNA) damage in individuals exposed to environmental stresses and chemical carcinogens is unknown. In the study, we investigated the association between Hsp70 levels (the most abundant mammalian Hsp) and genotoxic damage in lymphocytes of workers exposed to coke-oven emission using Western dot blot an...

  2. Transcription factor TFIID recognizes DNA sequences downstream of the TATA element in the Hsp70 heat shock gene.

    OpenAIRE

    Emanuel, P A; Gilmour, D S

    1993-01-01

    The interaction between the Hsp70 heat shock gene promoter and a Drosophila protein complex which contains the TATA-binding protein depends on sequence-specific interactions located in the region downstream of the transcription start site. Immunopurification of the complex through the use of antibodies against the TATA-binding protein reveals that the complex is transcription factor TFIID. Binding assays with the immunopurified TFIID confirm that sequence-specific contacts are made in the reg...

  3. Fusion of Hsp70 to Mage-a1 enhances the potency of vaccine-specific immune responses

    OpenAIRE

    Jiang, Juhong; Xie, Dan; Zhang, WenMin; Xiao, Gang; Wen, Jianming

    2013-01-01

    Background Heat shock proteins (HSPs) are capable of promoting antigen presentation of chaperoned peptides through interactions with receptors on antigen presenting cells. This property of HSPs suggests a potential function as an adjuvant-free carrier to stimulate immune responses against a covalently linked fusion partner. MAGE-A1 is a likely candidate for tumor immunotherapy due to its abundant immunogenic epitopes and strict tumor specificity. To analyze the influence of HSP70 conjugation ...

  4. Comparative Study on the Immunogenicity between Hsp70 DNA Vaccine and Hsp65 DNA Vaccine in Human Mycobacterium Tuberculosis

    Institute of Scientific and Technical Information of China (English)

    DAI; Wuxing; HUANG; Hailang; YUAN; Ye; HU; Jiajie; HUANGFU; Yongmu

    2001-01-01

    The BALB/c mice were immunized with Hsp70 DNA and Hsp65 DNA vaccines in human Mycobacterium tuberculosis. Eight weeks after immunization, the eyeballs were removed, blood and spleen taken, and intraperitoneal macrophages were harvested. The lymphocytic stimulating index(SI) was used to measure the cellular proliferating ability and NO release to measure the phagocytic activity of the macrophages. With ELISA kit, the levels of interleukin-2 (IL-2) and interferon-γ(IFN-γ) in serum and the splenic lymphocytic cultured supernatant were detected. The results showed that after the mice were immunized with 100 μg/mouse of Hsp70 DNA vaccine intramuscularly, the splenic lymphocytic proliferating ability in the mice was significantly increased as compared with that in the control group, vector group and Hsp65 DNA vaccine group (P<0. 01); The contents of NO in the intraperitoneal macrophages of the mice were significantly lower than in the control group and Hsp65 DNA vaccine group (P<0. 01); The levels of serum IL-2 in the mice were significantly higher than in the control group, but there was no statistical difference between Hsp65 DNA group and vector group (P>0. 05); The contents of serum IFN-γ in the mice were significantly higher than in the control group, but significantly lower than in the Hsp65 DNA vaccine group (P<0. 05). It was indicated that immunization with Hsp70 DNA vaccine could obviously enhance the immune response, but its intensity seemed inferior to Hsp65 DNA vaccine. The anti-infection mechanisms and clinical use in the future of the vaccines of Hsp70 DNA and Hsp65 DNA are worth further studying.

  5. Function of SSA subfamily of Hsp70 within and across species varies widely in complementing Saccharomyces cerevisiae cell growth and prion propagation.

    Directory of Open Access Journals (Sweden)

    Deepak Sharma

    Full Text Available BACKGROUND: The cytosol of most eukaryotic cells contains multiple highly conserved Hsp70 orthologs that differ mainly by their spatio-temporal expression patterns. Hsp70s play essential roles in protein folding, transport or degradation, and are major players of cellular quality control processes. However, while several reports suggest that specialized functions of Hsp70 orthologs were selected through evolution, few studies addressed systematically this issue. METHODOLOGY/PRINCIPAL FINDINGS: We compared the ability of Ssa1p-Ssa4p from Saccharomyces cerevisiae and Ssa5p-Ssa8p from the evolutionary distant yeast Yarrowia lipolytica to perform Hsp70-dependent tasks when expressed as the sole Hsp70 for S. cerevisiae in vivo. We show that Hsp70 isoforms (i supported yeast viability yet with markedly different growth rates, (ii influenced the propagation and stability of the [PSI(+] and [URE3] prions, but iii did not significantly affect the proteasomal degradation rate of CFTR. Additionally, we show that individual Hsp70 orthologs did not induce the formation of different prion strains, but rather influenced the aggregation properties of Sup35 in vivo. Finally, we show that [URE3] curing by the overexpression of Ydj1p is Hsp70-isoform dependent. CONCLUSION/SIGNIFICANCE: Despite very high homology and overlapping functions, the different Hsp70 orthologs have evolved to possess distinct activities that are required to cope with different types of substrates or stress situations. Yeast prions provide a very sensitive model to uncover this functional specialization and to explore the intricate network of chaperone/co-chaperone/substrates interactions.

  6. Effect of neoadjuvant chemoradiation and postoperative radiotherapy on expression of heat shock protein 70 (HSP70) in head and neck vessels

    International Nuclear Information System (INIS)

    Preoperative radiotherapy and chemotherapy in patients with head and neck cancer result in changes to the vessels that are used to construct microsurgical anastomoses. The aim of the study was to investigate quantitative changes and HSP70 expression of irradiated neck recipient vessels and transplant vessels used for microsurgical anastomoses. Of 20 patients included in this study five patients received neoadjuvant chemoradiation, another five received conventional radiotherapy and 10 patients where treated without previous radiotherapy. During surgical procedure, vessel specimens where obtained by the surgeon. Immunhistochemical staining of HSP70 was performed and quantitative measurement and evaluation of HSP70 was carried out. Conventional radiation and neoadjuvant chemoradiation revealed in a thickening of the intima layer of recipient vessels. A increased expression of HSP70 could be detected in the media layer of the recipient veins as well as in the transplant veins of patients treated with neoadjuvant chemoradiation. Radiation and chemoradiation decreased the HSP70 expression of the intima layer in recipient arteries. Conventional radiation led to a decrease of HSP70 expression in the media layer of recipient arteries. Our results showed that anticancer drugs can lead to a thickening of the intima layer of transplant and recipient veins and also increase the HSP70 expression in the media layer of the recipient vessels. In contrast, conventional radiation decreased the HSP70 expression in the intima layer of arteries and the media layer of recipient arteries and veins. Comparing these results with wall thickness, it was concluded, that high levels of HSP70 may prevent the intima layer of arteries and the media layer of vein from thickening

  7. Effects of heated hydrotherapy on muscle HSP70 and glucose metabolism in old and young vervet monkeys.

    Science.gov (United States)

    Kavanagh, Kylie; Davis, Ashely T; Jenkins, Kurt A; Flynn, D Mickey

    2016-07-01

    Increasing heat shock protein 70 (HSP70) in aged and/or insulin-resistant animal models confers benefits to healthspan and lifespan. Heat application to increase core temperature induces HSPs in metabolically important tissues, and preliminary human and animal data suggest that heated hydrotherapy is an effective method to achieve increased HSPs. However, safety concerns exist, particularly in geriatric medicine where organ and cardiovascular disease commonly will preexist. We evaluated young vervet monkeys compared to old, insulin-resistant vervet monkeys (Chlorocebus aethiops sabaeus) in their core temperatures, glucose tolerance, muscle HSP70 level, and selected safety biomarkers after 10 sessions of hot water immersions administered twice weekly. Hot water immersion robustly induced the heat shock response in muscles. We observed that heat-treated old and young monkeys have significantly higher muscle HSP70 than control monkeys and treatment was without significant adverse effects on organ or cardiovascular health. Heat therapy improved pancreatic responses to glucose challenge and tended to normalize glucose excursions. A trend for worsened blood pressure and glucose values in the control monkeys and improved values in heat-treated monkeys were seen to support further investigation into the safety and efficacy of this intervention for metabolic syndrome or diabetes in young or old persons unable to exercise. PMID:27188431

  8. Survival of human epidermal keratinocytes after short-duration high temperature: synthesis of HSP70 and IL-8.

    Science.gov (United States)

    Bowman, P D; Schuschereba, S T; Lawlor, D F; Gilligan, G R; Mata, J R; DeBaere, D R

    1997-06-01

    Thermal injury by short pulses (1-30 s) of relatively high temperature (50-68 degrees C) was investigated in normal human epidermal keratinocytes (NHEK). NHEK were cultured on plastic cover-slips and dipped in medium held at various temperatures. Survival assessed by methylthiazol tetrazolium reduction assay at 6 days postheating demonstrated an inverse time-temperature relationship that indicated that most cells could survive after a 1-s, 60 degrees C exposure or a 30-s, 55 degrees C exposure. Arrhenius plots of the data indicated major transition points for cell injury at 50 and 60 degrees C. Heat shock protein 70 (HSP70) and interleukin-8 (IL-8) were both induced by elevation of temperature between 50 and 60 degrees C for as short a time as 1 s. HSP70 synthesis stimulated by short, high pulses of heat appeared to induce thermotolerance. These results demonstrate that brief exposure to relatively high temperature can induce HSP70 and IL-8 synthesis in keratinocytes. PMID:9227428

  9. Mesothelioma Cells Escape Heat Stress by Upregulating Hsp40/Hsp70 Expression via Mitogen-Activated Protein Kinases

    Directory of Open Access Journals (Sweden)

    Michael Roth

    2009-01-01

    Full Text Available Therapy with hyperthermal chemotherapy in pleural diffuse malignant mesothelioma had limited benefits for patients. Here we investigated the effect of heat stress on heat shock proteins (HSP, which rescue tumour cells from apoptosis. In human mesothelioma and mesothelial cells heat stress (39–42°C induced the phosphorylation of two mitogen activated kinases (MAPK Erk1/2 and p38, and increased Hsp40, and Hsp70 expression. Mesothelioma cells expressed more Hsp40 and were less sensitive to heat stress compared to mesothelial cells. Inhibition of Erk1/2 MAPK by PD98059 or by Erk1 siRNA down-regulated heat stress-induced Hsp40 and Hsp70 expression and reduced mesothelioma cell survival. Inhibition of p38MAPK by SB203580 or siRNA reduced Hsp40, but not Hsp70, expression and also increased mesothelioma cell death. Thus hyperthermia combined with suppression of p38 MAPK or Hsp40 may represent a novel approach to improve mesothelioma therapy.

  10. Schiff Base Metal Derivatives Enhance the Expression of HSP70 and Suppress BAX Proteins in Prevention of Acute Gastric Lesion

    Directory of Open Access Journals (Sweden)

    Shahram Golbabapour

    2013-01-01

    Full Text Available Schiff base complexes have appeared to be promising in the treatment of different diseases and disorders and have drawn a lot of attention to their biological activities. This study was conducted to evaluate the regulatory effect of Schiff base metal derivatives on the expression of heat shock proteins (HSP 70 and BAX in protection against acute haemorrhagic gastric ulcer in rats. Rats were assigned to 6 groups of 6 rats: the normal control (Tween 20 5% v/v, 5 mL/kg, the positive control (Tween 20 5% v/v, 5 mL/kg, and four Schiff base derivative groups named Schiff_1, Schiff_2, Schiff_3, and Schiff_4 (25 mg/kg. After 1 h, all of the groups received ethanol 95% (5 mL/kg but the normal control received Tween 20 (Tween 20 5% v/v, 5 mL/kg. The animals were euthanized after 60 min and the stomachs were dissected for histology (H&E, immunohistochemistry, and western blot analysis against HSP70 and BAX proteins. The results showed that the Schiff base metal derivatives enhanced the expression of HSP70 and suppressed the expression of BAX proteins during their gastroprotection against ethanol-induced gastric lesion in rats.

  11. Correlation of Hsp70 Serum Levels with Gross Tumor Volume and Composition of Lymphocyte Subpopulations in Patients with Squamous Cell and Adeno Non-Small Cell Lung Cancer.

    Science.gov (United States)

    Gunther, Sophie; Ostheimer, Christian; Stangl, Stefan; Specht, Hanno M; Mozes, Petra; Jesinghaus, Moritz; Vordermark, Dirk; Combs, Stephanie E; Peltz, Friedhelm; Jung, Max P; Multhoff, Gabriele

    2015-01-01

    Heat-shock protein 70 (Hsp70) is frequently found on the plasma membrane of a large number of malignant tumors including non-small cell lung cancer (NSCLC) and gets released into the blood circulation in lipid vesicles. On the one hand, a membrane (m)Hsp70-positive phenotype correlates with a high aggressiveness of the tumor; on the other hand, mHsp70 serves as a target for natural killer (NK) cells that had been pre-stimulated with Hsp70-peptide TKD plus low-dose interleukin-2 (TKD/IL-2). Following activation, NK cells show an up-regulated expression of activatory C-type lectin receptors, such as CD94/NKG2C, NKG2D, and natural cytotoxicity receptors (NCRs; NKp44, NKp46, and NKp30) and thereby gain the capacity to kill mHsp70-positive tumor cells. With respect to these results, the efficacy of ex vivo TKD/IL-2 stimulated, autologous NK cells is currently tested in a proof-of-concept phase II clinical trial in patients with squamous cell NSCLC after radiochemotherapy (RCT) at the TUM. Inclusion criteria are histological proven, non-resectable NSCLC in stage IIIA/IIIB, clinical responses to RCT and a mHsp70-positive tumor phenotype. The mHsp70 status is determined in the serum of patients using the lipHsp70 ELISA test, which enables the quantification of liposomal and free Hsp70. Squamous cell and adeno NSCLC patients had significantly higher serum Hsp70 levels than healthy controls. A significant correlation of serum Hsp70 levels with the gross tumor volume was shown for adeno and squamous cell NSCLC. However, significantly elevated ratios of activated CD69(+)/CD94(+) NK cells that are associated with low serum Hsp70 levels were observed only in patients with squamous cell lung cancer. These data might provide a first hint that squamous cell NSCLC is more immunogenic than adeno NSCLC. PMID:26579130

  12. Conserved Distal Loop Residues in the Hsp104 and ClpB Middle Domain Contact Nucleotide-binding Domain 2 and Enable Hsp70-dependent Protein Disaggregation*

    OpenAIRE

    DeSantis, Morgan E.; Sweeny, Elizabeth A.; Snead, David; Leung, Eunice H.; Go, Michelle S.; Gupta, Kushol; Wendler, Petra; Shorter, James

    2013-01-01

    The homologous hexameric AAA+ proteins, Hsp104 from yeast and ClpB from bacteria, collaborate with Hsp70 to dissolve disordered protein aggregates but employ distinct mechanisms of intersubunit collaboration. How Hsp104 and ClpB coordinate polypeptide handover with Hsp70 is not understood. Here, we define conserved distal loop residues between middle domain (MD) helix 1 and 2 that are unexpectedly critical for Hsp104 and ClpB collaboration with Hsp70. Surprisingly, the Hsp104 and ClpB MD dist...

  13. Multifunctional selenium nanoparticles as carriers of HSP70 siRNA to induce apoptosis of HepG2 cells

    Directory of Open Access Journals (Sweden)

    Li Y

    2016-07-01

    Full Text Available Yinghua Li,1 Zhengfang Lin,1 Mingqi Zhao,1 Tiantian Xu,1 Changbing Wang,1 Huimin Xia,1,* Hanzhong Wang,2,* Bing Zhu1,* 1Guangzhou Women and Children’s Medical Center, Guangzhou, Guangdong, 2State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People’s Republic of China *These authors contributed equally to this work Abstract: Small interfering RNA (siRNA as a new therapeutic modality holds promise for cancer treatment, but it is unable to cross cell membrane. To overcome this limitation, nanotechnology has been proposed for mediation of siRNA transfection. Selenium (Se is a vital dietary trace element for mammalian life and plays an essential role in the growth and functioning of humans. As a novel Se species, Se nanoparticles have attracted more and more attention for their higher anticancer efficacy. In the present study, siRNAs with polyethylenimine (PEI-modified Se nanoparticles (Se@PEI@siRNA have been demonstrated to enhance the apoptosis of HepG2 cells. Heat shock protein (HSP-70 is overexpressed in many types of human cancer and plays a significant role in several biological processes including the regulation of apoptosis. The objective of this study was to silence inducible HSP70 and promote the apoptosis of Se-induced HepG2 cells. Se@PEI@siRNA were successfully prepared and characterized by various microscopic methods. Se@PEI@siRNA showed satisfactory size distribution, high stability, and selectivity between cancer and normal cells. The cytotoxicity of Se@PEI@siRNA was lower for normal cells than tumor cells, indicating that these compounds may have fewer side effects. The gene-silencing efficiency of Se@PEI@siRNA was significantly much higher than Lipofectamine 2000@siRNA and resulted in a significantly reduced HSP70 mRNA and protein expression in cancer cells. When the expression of HSP70 was diminished, the function of cell protection was also removed and cancer cells became more

  14. GENE EXPRESSION PROFILES IN ARSENIC-TREATED MCF-7 BREAST CANCER CELLS EXPRESSING DIFFERENT LEVELS OF HSP70

    Science.gov (United States)

    Gene expression profiles in arsenic-treated MCF-7 breast cancer cells expressing different levels of HSP70Gail Nelson, Susan Hester, Ernest Winkfield, Jill Barnes, James AllenEnvironmental Carcinogenesis Division, NHEERL, ORD, US Environmental Protection Agency, Rese...

  15. The expressions of HSP70 and αB-crystallin in myocarditis associated with foot-and-mouth disease virus in lambs

    OpenAIRE

    Gulbahar, Mustafa Yavuz; Kabak, Yonca Betil; Karayigit, Mehmet Onder; Yarim, Murat; Guvenc, Tolga; Parlak, Unal

    2011-01-01

    This study describes the expression of heat shock protein70 (HSP70) and alpha-basic-crystallin (α-BC) and their association with apoptosis and some related adaptor proteins in the pathogenesis of foot-and-mouth disease virus (FMDV)-induced myocarditis in lambs. HSP70 was generally overexpressed in the myocardial tissues and inflammatory cells of FMDV-induced myocarditis with differential accumulation and localization in same hearts when compared to non-foot-and-mouth disease control hearts. α...

  16. TATA-dependent and TATA-independent function of the basal and heat shock elements of a human hsp70 promoter.

    OpenAIRE

    Greene, J M; Kingston, R.E.

    1990-01-01

    We have characterized the interactions between the TATA element and other sequence elements of a human heat shock protein 70 (hsp70) promoter by a mutational approach. Expression of a distal element of this promoter requires an intact TATA element in human cell lines. The hsp70 TATA element can be functionally replaced for this interaction by TATA elements from the simian virus 40 early and adenovirus EIIa promoters. The TATA element in this promoter therefore both determines the appropriate ...

  17. Non-lethal heat shock increased Hsp70 and immune protein transcripts but not Vibrio tolerance in the white-leg shrimp.

    Directory of Open Access Journals (Sweden)

    Nguyen Hong Loc

    Full Text Available Non-lethal heat shock boosts bacterial and viral disease tolerance in shrimp, possibly due to increases in endogenous heat shock protein 70 (Hsp70 and/or immune proteins. To further understand the mechanisms protecting shrimp against infection, Hsp70 and the mRNAs encoding the immune-related proteins prophenoloxidase (proPO, peroxinectin, penaeidin, crustin and hemocyanin were studied in post-larvae of the white-leg shrimp Litopenaeus vannamei, following a non-lethal heat shock. As indicated by RT-qPCR, a 30 min abrupt heat shock increased Hsp70 mRNA in comparison to non-heated animals. Immunoprobing of western blots and quantification by ELISA revealed that Hsp70 production after heat shock was correlated with enhanced Hsp70 mRNA. proPO and hemocyanin mRNA levels were augmented, whereas peroxinectin and crustin mRNA levels were unchanged following non-lethal heat shock. Penaeidin mRNA was decreased by all heat shock treatments. Thirty min abrupt heat shock failed to improve survival of post-larvae in a standardized challenge test with Vibrio harveyi, indicating that under the conditions of this study, L. vannamei tolerance to Vibrio infection was influenced neither by Hsp70 accumulation nor the changes in the immune-related proteins, observations dissimilar to other shrimp species examined.

  18. Radiation combined with hyperthermia induces HSP70-dependent maturation of dendritic cells and release of pro-inflammatory cytokines by dendritic cells and macrophages

    International Nuclear Information System (INIS)

    Purpose: Hyperthermia (HT) treatment of cancer patients was revived over the last years and has been proven to be beneficiary for many cancer entities when applied temperature controlled in multimodal treatments. We examined whether a combination of ionizing irradiation (X-ray) and HT (41.5 °C; 1 h) can induce the release of heat shock protein (HSP) 70 by tumor cells and thereby lead to the activation of dendritic cells and macrophages. Material and methods: Extracellular HSP70 was detected in supernatants (SN) of treated colorectal tumor cells by ELISA. Maturation of dendritic cells (DC) after contact with the SN was measured by flow-cytometry. Phagocytosis assays were conducted to get hints about the immune stimulating potential of the tumor cells after the respective treatments. Results: An increased surface expression of HSP70 was observed after X-ray or X-ray plus HT while the amount of extracellular HSP70 was only increased when HT was given additionally. A high up-regulation of the co-stimulation molecule CD80 and the chemokine receptor CCR7 on DC was measured after contact with SN of X-ray plus HT treated cells. This was dependent on extracellular HSP70. Combined treatments further led to significantly increased phagocytosis rates of macrophages and DC and increased pro-inflammatory cytokine (IL-8 and IL-12) secretion. Conclusion: X-ray combined with HT induces HSP70 dependent activation of immune cells and might generate a tumor microenvironment beneficial for cure.

  19. Evaluation of immune effect of recombinant fusion protein targeting the prostate stem cell antigen based on PSCA and HSP70

    Directory of Open Access Journals (Sweden)

    Lei DONG

    2014-10-01

    Full Text Available Objective To explore the immune effect and antitumor activity of recombinant prostate stem cell protein (PSCA and heat shock protein 70 (HSP70 in a murine model of prostate cancer. Methods Twenty-five healthy male C57BL/6 mice were randomly divided into 5 groups (5 each: PSCA, HSP, PSCA+HSP, PSCA-HSP and control group. Mice in the first 4 groups were vaccinated with the corresponding proteins, and those in control group were faked with injection of phosphate buffer saline (PBS. After immunization with recombinant proteins, the PSCA-specific cellular immune responses were monitored with ELISPOT, intracellular cytokine staining assay, and flow cytometry, and ELISA assay was used to detect humoral immune responses. The tumor growth and survival of vaccined mice were observed. Results ELISPOT revealed that the mice in PSCA-HSP group generated much more IFN-γ spot-forming cells than those in other groups (P<0.05, and they could generate strong anti-PSCA antibody response. Results of flow cytometry showed that the number of CD8+/IFN-γ+ T cells was significantly higher in PSCAHSP group than that in other groups (P<0.05. ELISA results revealed that all the mice in PSCA, PSCA+HSP and PSCA-HSP group were induced to generate the PSCA-specific humoral immune response, and no statistical difference was found on the antibody levels among the three groups. Animal experiment showed that PSCA-HSP could inhibit the growth of PSCA-expressing tumors and prolong the survival time of vaccinated mice. Conclusion HSP70 is a chaperone with significant effect for protein vaccines, and the recombinant fusion protein PSCA-HSP70 could be of potential value for prostate cancer treatment. DOI: 10.11855/j.issn.0577-7402.2014.09.08

  20. 17-DMAG, an HSP90 Inhibitor, Ameliorates Multiple Organ Dysfunction Syndrome via Induction of HSP70 in Endotoxemic Rats.

    Directory of Open Access Journals (Sweden)

    Yi-Li Wang

    Full Text Available Sepsis is a systemic inflammatory disorder, accompanied with elevated oxidative stress, leading to multiple organ dysfunction syndrome (MODS, and disseminated intravascular coagulation. 17-Dimethylaminoethylamino- 17-demethoxygeldanamycin (17-DMAG, a heat shock protein (HSP 90 inhibitor, has been reported to possess anti-inflammatory effects. In this study, the beneficial effects of 17-DMAG on lipopolysaccharide (LPS induced MODS and DIC was evaluated in anesthetized rats. 17-DMAG (5 mg/kg, i.p. was significantly increased survival rate, and prevented hypotension in LPS (30 mg/kg i.v. infused for 4 h induced endotoxemia. The elevated levels of alanine aminotransferase (ALT, creatine phosphokinase (CPK, lactate dehydrogenase, creatinine, nitric oxide (NO metabolites, IL-6, and TNF-α in LPS-exposed rat plasma were significantly reduced by 17-DMAG. Moreover, 17-DMAG suppressed LPS-induced superoxide anion production and caspase 3 activation in heart tissues. LPS induced the prolongation of prothrombin time, and a pronounced decrease in platelet count, which were improved by 17-DMAG. 17-DMAG markedly induced HSP70 and heme oxygenase (HO-1, and suppressed inducible nitric oxide synthase (iNOS and phosphorylated NF-κB p65 protein expression in organs 6 h after LPS initiation. Pretreatment with high dose of quercetin (300 mg/kg, i.p., as an HSP70 inhibitor, reversed the beneficial effects of 17-DMAG on survival rate, plasma levels of ALT, CPK, creatinine, IL-6, and NO metabolites, iNOS induction, and caspase-3 activation in LPS-treated rats. In conclusion, 17-DMAG possesses the anti-inflammatory and antioxidant effects that were proved through LPS-induced acute inflammation, which is associated with induction of HSP70 and HO-1, leading to prevent MODS in sepsis.

  1. 17-DMAG, an HSP90 Inhibitor, Ameliorates Multiple Organ Dysfunction Syndrome via Induction of HSP70 in Endotoxemic Rats.

    Science.gov (United States)

    Wang, Yi-Li; Shen, Hsin-Hsueh; Cheng, Pao-Yun; Chu, Yen-Ju; Hwang, Hwong-Ru; Lam, Kwok-Keung; Lee, Yen-Mei

    2016-01-01

    Sepsis is a systemic inflammatory disorder, accompanied with elevated oxidative stress, leading to multiple organ dysfunction syndrome (MODS), and disseminated intravascular coagulation. 17-Dimethylaminoethylamino- 17-demethoxygeldanamycin (17-DMAG), a heat shock protein (HSP) 90 inhibitor, has been reported to possess anti-inflammatory effects. In this study, the beneficial effects of 17-DMAG on lipopolysaccharide (LPS) induced MODS and DIC was evaluated in anesthetized rats. 17-DMAG (5 mg/kg, i.p.) was significantly increased survival rate, and prevented hypotension in LPS (30 mg/kg i.v. infused for 4 h) induced endotoxemia. The elevated levels of alanine aminotransferase (ALT), creatine phosphokinase (CPK), lactate dehydrogenase, creatinine, nitric oxide (NO) metabolites, IL-6, and TNF-α in LPS-exposed rat plasma were significantly reduced by 17-DMAG. Moreover, 17-DMAG suppressed LPS-induced superoxide anion production and caspase 3 activation in heart tissues. LPS induced the prolongation of prothrombin time, and a pronounced decrease in platelet count, which were improved by 17-DMAG. 17-DMAG markedly induced HSP70 and heme oxygenase (HO)-1, and suppressed inducible nitric oxide synthase (iNOS) and phosphorylated NF-κB p65 protein expression in organs 6 h after LPS initiation. Pretreatment with high dose of quercetin (300 mg/kg, i.p.), as an HSP70 inhibitor, reversed the beneficial effects of 17-DMAG on survival rate, plasma levels of ALT, CPK, creatinine, IL-6, and NO metabolites, iNOS induction, and caspase-3 activation in LPS-treated rats. In conclusion, 17-DMAG possesses the anti-inflammatory and antioxidant effects that were proved through LPS-induced acute inflammation, which is associated with induction of HSP70 and HO-1, leading to prevent MODS in sepsis. PMID:27224288

  2. Expression of Hsp27 and Hsp70 in Lymphocytes and Plasma in Healthy Workers and Coal Miners with Lung Cancer

    Institute of Scientific and Technical Information of China (English)

    王海椒; 邢景才; 王峰; 韩文慧; 任侯卯; 邬堂春; 陈卫红

    2010-01-01

    In coal mines, main occupational hazard is coal-mine dust, which can cause health problem including coal workers' pneumoconiosis and lung cancer. Some heat shock proteins (Hsps) have been reported as an acute response to a wide variety of stressful stimuli. Whether Hsps protect against chronic environmental coal-mine dust over years is unknown. It is also interesting to know that whether the expression of Hsp27 and Hsp70 proteins as a marker for exposure is associated risk of lung cancer among coal miners. ...

  3. Comparative evaluation of recombinant HSP70 (N & C-terminal) fragments in the detection of equine trypanosomosis.

    Science.gov (United States)

    Kumar, Jaideep; Chaudhury, A; Yadav, S C

    2016-06-15

    Trypanosomosis (Surra) is an economically important disease caused by Trypanosoma evansi which is an extracellular parasite present in the plasma, tissues and other body fluids of a wide range of hosts including domesticated animals. Currently, serological reports are based on detection of antibodies by ELISA using whole cell lysate (WCL) antigen, which has a limitation of persistence of anti-trypanosomal antibodies after successful treatment of the disease. Moreover, it has some ethical issues also like requirement of mice for in vivo maintenance of parasite for preparing the antigen. Therefore, in the present study, an attempt was made to evaluate the in vitro production of recombinant heat shock protein 70 (HSP70) for detection of antibodies in experimentally infected ponies. The amino acid sequence analysis of HSP70 revealed that N-terminal region of the protein was highly conserved while the C-terminal region was most divergent. The four different regions of HSP70 protein viz. HSP-1, HSP-2, HSP-3 and HSP-4 were cloned and expressed, among which HSP-1 (N-terminal region) & HSP-2 (C-terminal region) were truncated while HSP-3 & HSP-4 were complete C-terminal proteins. The recombinant fragments were probed with sequentially pooled experimental serum samples where antibodies were detected in these fragments from 10(th) day post infection till the termination of the experiment. Further, these recombinant fragments were also comparatively evaluated with WCL antigen in ELISA using experimental as well as field serum samples. It was observed that after successful treatment of infected ponies, there was a sharp fall in antibodies (within 90 days) when tested with recombinant HSP's fragments, while antibodies persisted even after 469 days when tested against WCL antigen. The sensitivity and specificity of all HSP70 fragments were also estimated from field serum samples with reference to WCL antigen ELISA. The HSP-1 showed minimum sensitivity (41.03%) among all the

  4. Evaluation of immune effect of recombinant fusion protein targeting the prostate stem cell antigen based on PSCA and HSP70

    OpenAIRE

    Dong, Lei; Liu, Juan; Ma, Hong-Yu; Shou-zhen QUAN; Jian-xiao LV; Ying LI; Zhu, Mei-Cai

    2014-01-01

    Objective To explore the immune effect and antitumor activity of recombinant prostate stem cell protein (PSCA) and heat shock protein 70 (HSP70) in a murine model of prostate cancer. Methods Twenty-five healthy male C57BL/6 mice were randomly divided into 5 groups (5 each): PSCA, HSP, PSCA+HSP, PSCA-HSP and control group. Mice in the first 4 groups were vaccinated with the corresponding proteins, and those in control group were faked with injection of phosphate buffer saline (PBS). After immu...

  5. Hsc66, an Hsp70 homolog in Escherichia coli, is induced by cold shock but not by heat shock.

    OpenAIRE

    Lelivelt, M J; Kawula, T H

    1995-01-01

    Hsc66 is the second identified Hsp70 protein in Escherichia coli. Mutations in hscA, the gene encoding Hsc66, compensate for some phenotypic effects of a mutation in hns, a gene encoding the cold-inducible, nucleoid-associated protein H-NS. Expression of hscA was not induced upon heat shock but was induced approximately 11-fold 3 h after a shift from 37 to 10 degrees C. Furthermore, hscA was induced upon chloramphenicol addition, which induces the synthesis of other cold-inducible genes. Mapp...

  6. Thermal stress in Larvae of the sand dollar (Dendraster excentricus) induces changes in hsp70 gen expression

    Science.gov (United States)

    Olivares-Bañuelos, T.; Garcia-Echauri, L.; Figueroa-Flores, S.; Carpizo-Ituarte, E.

    2010-03-01

    The relationship between temperature and individual performance is reasonably well understood, and much climate-related research has focused on potential shifts in distribution and abundance driven directly by temperature(1). It is known, that global climate change has profound implications for marine ecosystems and the economic and social systems that depend upon them. In virtually all organisms, including equinoderms, cells respond to a variety of stresses, as temperature changes, by the rapid synthesis of a highly conserved set of polypeptides termed heat shock proteins (HSPs) (2). Differential Hsp expression among species may allow functionally important genetic variation to cope with thermal stress and other environmental stressors, allowing populations to respond effectively to environmental change. The aim of the present work was to elucidate the effect of different temperatures on hsp70 expression in larvae of the sand dollar Dendraster excentricus. We are interested to understand what is how organisms that live in estuaries respond to global warming. We did thermo tolerance assays whit 3 different larval stages of the sand dollar, including 8 arms plutei, competent and metamorphic larvae. We tested 4 experimental temperatures (4, 20, 26, and 29°C) and 3 different times (1, 3 and 6 hours). After each treatment, total RNA was extracted and used to quantify hsp70 expression levels using real time PCR analysis. We detected, after 6 hours of thermal stress, the highest over-expression levels of hsp70 in both metamorphic and competent larvae. In 8 arms pluetus larvae, no significant differences in hsp70 expression were observed after the exposure to the experimental temperatures. Taking together our results, indicate that larvae of Dendraster excentricus are responding according to their development stage to cope with thermal-stress. The capability of sand dollar larvae to acclimate and respond effectively to the climatic change would be determinated by the sea

  7. Modulation of Akt and ERK1/2 pathways by resveratrol in chronic myelogenous leukemia (CML cells results in the downregulation of Hsp70.

    Directory of Open Access Journals (Sweden)

    Soumyajit Banerjee Mustafi

    Full Text Available BACKGROUND: Resveratrol is known to downregulate the high endogenous level of Heat shock protein 70 (Hsp70 in Chronic Myelogenous Leukemia (CML K562 cells and induce apoptosis. Since Heat Shock Factor 1 (HSF1 controls transcription of Hsp70, we wanted to probe the signaling pathways responsible for transcriptional activation of HSF1. METHODOLOGY/PRINCIPAL FINDINGS: Cells exposed to 40microM Resveratrol rapidly abolished serine473 phosphorylation of Akt and significantly reduced its kinase activity. Inactivation of Akt pathway by Resveratrol subsequently blocked serine9 phosphorylation of Gsk3beta. Active non-phosphorylated Gsk3beta rendered HSF1 transcriptionally inactive and reduced Hsp70 production. Blocking PI3K/Akt activity also demonstrated similar effects on Hsp70 comparable to Resveratrol. Inactivation of Gsk3beta activity by inhibitors SB261763 or LiCl upregulated Hsp70. Resveratrol significantly modulated ERK1/2 activity as evident from hyper phosphorylation at T302/Y304 residues and simultaneous upregulation in kinase activity. Blocking ERK1/2 activation resulted in induction of Hsp70. Therefore, increase in ERK1/2 activity by Resveratrol provided another negative influence on Hsp70 levels through negative regulation of HSF1 activity. 17-allylamino-17-demethoxygeldanamycin (17AAG, a drug that inhibits Hsp90 chaperone and degrades its client protein Akt concomitantly elevated Hsp70 levels by promoting nuclear translocation of HSF1 from the cytosol. This effect is predominantly due to inhibition of both Akt and ERK1/2 activation by 17AAG. Simultaneously treating K562 with Resveratrol and 17AAG maintained phosho-ERK1/2 levels close to untreated controls demonstrating their opposite effects on ERK1/2 pathway. Resveratrol was found not to interfere with Bcr-Abl activation in K562 cells. CONCLUSION/SIGNIFICANCE: Thus our study comprehensively illustrates that Resveratrol acts downstream of Bcr-Abl and inhibits Akt activity but stimulates ERK

  8. In vitro Study on Role of Hsp70 Expression in DNA Damage of Human Embryonic Lung Cells Exposed to Benzo[a]pyrene

    Institute of Scientific and Technical Information of China (English)

    YA-JUAN GAO; CHENG-FENG XIAO; SHENG CHEN; RUI-BO WANG; HAN-ZHEN HE; ROBERT M TANGUAY; TANG-CHUN WU

    2004-01-01

    Objective Benzo[a]pyrene (B[a]P), a ubiquitous environmental pollutant, is a potent procarcinogen and mutagen that can elicit tumors, leading to malignancy. Heat shock proteins (Hsp) have been shown to protect cells against damages caused by various stresses including exposure to numerous chemicals. Whether Hsps, or more specifically Hsp70, are involved in repair of B[a]P-induced DNA damage is currently unknown. Methods We assessed the potential role of the inducible form of Hsp70 in B[a]P-induced DNA damage of human embryonic lung (HEL) cells using immunoblot and the comet assay (i.e., the single cell gel electrophoresis assay). Results Exposure to B[a]P induced a dose-dependent decrease in the level of Hsp70, but a dose-dependent increase in DNA damage both in untreated (control) HEL cells and in cells preconditioned by a heat treatment. Heat preconditioning prior to B[a]P exposure potentiated the effect of B[a]P at a low dose (10 (mol/L), but appeared to be protective at higher doses. There was a negative correlation between Hsp70 level and DNA damage in the non-preheated as well as in the preconditioned cells. Conclusion These data suggest that exposure of HEL cells to B[a]P may induce a dose-dependent reduction in the levels of the inducible Hsp70. The detailed mechanisms for the reduction of Hsp70 levels by B[a]P and the role of Hsp70 in DNA damage under different concentrations of B[a]P remains to be determined.

  9. Mild heat stress at a young age in Drosophila melanogaster leads to increased Hsp70 synthesis after stress exposure later in life

    Indian Academy of Sciences (India)

    Torsten Nygaard Kristensen; Jesper Givskov Sørensen; Volker Loeschcke

    2003-12-01

    In a number of animal species it has been shown that exposure to low levels of stress at a young age has a positive effect on stress resistance later in life, and on longevity. The positive effects have been attributed to the activation of defence/cleaning systems (heat shock proteins (Hsps), antioxidases, DNA repair) or to effects of a changed metabolic rate, or both. We investigated the effect of mild stress exposures early in life on Hsp70 synthesis after a harder stress exposure later in life in five isofemale lines of Drosophila melanogaster. Female flies were either exposed to repeated bouts of mild heat stress (3 h at 34°C) at a young age (days 2, 4 and 6 post-eclosion) or held under standard laboratory conditions. At 16 and 32 days of adult age, respectively, flies were exposed to a high temperature treatment known to induce Hsp70 in the investigated species (1 h at 37°ºC). Thereafter, the inducible Hsp70 levels were measured. Our data show a tendency towards increased Hsp70 synthesis with increased age for both ‘mild stress’ and ‘no stress’ flies. Moreover, the results show that flies exposed to mild stress at a young age synthesized more Hsp70 upon induction, compared to control flies, and that this difference was accentuated at 32 days compared to 16 days of age. Thus, bouts of mild heat stress at a young age impact on the physiological stress response system later in life. This may be caused by an increased ability to react to future stresses. Alternatively, the mild stress exposure at a young age may actually have caused cellular damages increasing the need for Hsp70 levels after stress exposure later in life. The importance of an Hsp70 upregulation (throughout life) in explaining the phenomenon of hormesis is discussed, together with alternative hypotheses, and suggestions for further studies.

  10. Cryo-thermal therapy elicits potent anti-tumor immunity by inducing extracellular Hsp70-dependent MDSC differentiation

    Science.gov (United States)

    Zhu, Jun; Zhang, Yan; Zhang, Aili; He, Kun; Liu, Ping; Xu, Lisa X.

    2016-06-01

    Achieving control of metastatic disease is a long-sought goal in cancer therapy. Treatments that encourage a patient’s own immune system are bringing new hopes in reaching such a goal. In clinic, local hyperthermia and cryoablation have been explored to induce anti-tumor immune responses against tumors. We have also developed a novel therapeutic modality of cryo-thermal treatment by alternating liquid nitrogen (LN2) cooling and radio frequency (RF) heating, and better therapeutic effect was achieved in treating metastatic cancer in animal model. In this study, we investigated the mechanism of systemic immune response elicited by cryo-thermal therapy. In the 4T1 murine mammary carcinoma model, we found that local cryo-thermal therapy resulted in a considerable reduction of distant lung metastases, and improved long-term survival. Moreover, results of tumor re-challenge experiments indicated generation of a strong tumor-specific immune memory after the local treatment of primary tumors. Our further study indicated that cryo-thermal therapy caused an elevated extracellular release of Hsp70. Subsequently, Hsp70 induced differentiation of MDSCs into mature DCs, contributing to the relief of MDSCs-mediated immunosuppression and ultimately the activation of strong anti-tumor immune response. Our findings reveal new insight into the mechanism of robust therapeutic effects of cryo-thermal therapy against metastatic cancers.

  11. Cryo-thermal therapy elicits potent anti-tumor immunity by inducing extracellular Hsp70-dependent MDSC differentiation.

    Science.gov (United States)

    Zhu, Jun; Zhang, Yan; Zhang, Aili; He, Kun; Liu, Ping; Xu, Lisa X

    2016-01-01

    Achieving control of metastatic disease is a long-sought goal in cancer therapy. Treatments that encourage a patient's own immune system are bringing new hopes in reaching such a goal. In clinic, local hyperthermia and cryoablation have been explored to induce anti-tumor immune responses against tumors. We have also developed a novel therapeutic modality of cryo-thermal treatment by alternating liquid nitrogen (LN2) cooling and radio frequency (RF) heating, and better therapeutic effect was achieved in treating metastatic cancer in animal model. In this study, we investigated the mechanism of systemic immune response elicited by cryo-thermal therapy. In the 4T1 murine mammary carcinoma model, we found that local cryo-thermal therapy resulted in a considerable reduction of distant lung metastases, and improved long-term survival. Moreover, results of tumor re-challenge experiments indicated generation of a strong tumor-specific immune memory after the local treatment of primary tumors. Our further study indicated that cryo-thermal therapy caused an elevated extracellular release of Hsp70. Subsequently, Hsp70 induced differentiation of MDSCs into mature DCs, contributing to the relief of MDSCs-mediated immunosuppression and ultimately the activation of strong anti-tumor immune response. Our findings reveal new insight into the mechanism of robust therapeutic effects of cryo-thermal therapy against metastatic cancers. PMID:27256519

  12. High Resolution Melting Analysis Targeting hsp70 as a Fast and Efficient Method for the Discrimination of Leishmania Species.

    Directory of Open Access Journals (Sweden)

    Ricardo Andrade Zampieri

    2016-02-01

    Full Text Available Protozoan parasites of the genus Leishmania cause a large spectrum of clinical manifestations known as Leishmaniases. These diseases are increasingly important public health problems in many countries both within and outside endemic regions. Thus, an accurate differential diagnosis is extremely relevant for understanding epidemiological profiles and for the administration of the best therapeutic protocol.Exploring the High Resolution Melting (HRM dissociation profiles of two amplicons using real time polymerase chain reaction (real-time PCR targeting heat-shock protein 70 coding gene (hsp70 revealed differences that allowed the discrimination of genomic DNA samples of eight Leishmania species found in the Americas, including Leishmania (Leishmania infantum chagasi, L. (L. amazonensis, L. (L. mexicana, L. (Viannia lainsoni, L. (V. braziliensis, L. (V. guyanensis, L. (V. naiffi and L. (V. shawi, and three species found in Eurasia and Africa, including L. (L. tropica, L. (L. donovani and L. (L. major. In addition, we tested DNA samples obtained from standard promastigote culture, naturally infected phlebotomines, experimentally infected mice and clinical human samples to validate the proposed protocol.HRM analysis of hsp70 amplicons is a fast and robust strategy that allowed for the detection and discrimination of all Leishmania species responsible for the Leishmaniases in Brazil and Eurasia/Africa with high sensitivity and accuracy. This method could detect less than one parasite per reaction, even in the presence of host DNA.

  13. Developmental and hyperthermia-induced expression of the heat shock proteins HSP60 and HSP70 in tissues of the housefly Musca domestica: an in vitro study

    Directory of Open Access Journals (Sweden)

    Sunita Sharma

    2007-01-01

    Full Text Available The expression pattern of two major chaperones, the heat shock proteins (HSPs HSP60 and HSP70 was studied in vitro in tissues of the housefly Musca domestica during larval and adult stages of development to identify their immunological relatives and understand their functional significance in normal cellular activities and during thermal stress. Fluorographs of labeled polypeptides and western blots demonstrated that both HSPs are expressed constitutively and heat-induced in all the larval and adult cell types examined. The pattern of whole tissue immunocytochemical staining using anti-HSP60 and anti-HSP70 antibodies corresponded well with the observations from western blots or fluorographs. In developing oocytes, both constitutive and heat inducible expression of HSP60 were regulated in an oocyte stage-specific manner. In unstressed ovaries the expression of these proteins was less pronounced in early stage oocytes (1st - 8th than at later stages (9th and onward. The heat shock, however, induced both HSP70 and HSP60 to a significantly high level in early stage oocytes (1st-8th as compared to their respective controls. Our findings indicate the involvement of the HSP60 and HSP70 proteins in the development, growth and differentiation of both somatic and germ line tissues. Furthermore, the enhanced co-expression of HSP70 and HSP60 upon heat shock in various larval and adult cell types suggests the possible role of HSP60 in thermoprotection.

  14. [Hsp70 Fused with the Envelope Glycoprotein E0 of Classical Swine Fever Virus Enhances Immune Responses in Balb/c Mice].

    Science.gov (United States)

    Xu, Qianqian; Zhang, Xiaomin; Jing, Jiao; Shi, Baojun; Wang, Shiqi; Zhou, Bin; Chen, Puyan

    2015-07-01

    Heat-shock protein (Hsp) 70 potentiates specific immune responses to some antigenic peptides fused to it. Here, the prokaryotic plasmids harboring the envelope glycoprotein E0 gene of classical swine fever virus (CSFV) and/or the Hsp70 gene of Haemophilus parasuis were constructed and expressed in Escherichia coli Rosseta 2(R2). The fusion proteins were then purified. Groups of Balb/c mice were immunized with these fusion proteins, respectively, and sera collected 7 days after the third immunization. Immune effects were determined via an enzyme-linked immunosorbent assay and flow cytometric analyses. E0-Hsp70 fusion protein and E0+Hsp70 mixture significantly improved the titer of E-specific antibody, levels of CD4+ and CD8+ T cells, and release of interferon-γ. These findings suggested that Hsp70 can significantly enhance the immune effects of the envelope glycoprotein E0 of CSFV, thereby laying the foundation of further application in pigs. PMID:26524908

  15. Expression of hsp70, hsp90 and hsf1 in the reef coral Acropora digitifera under prospective acidified conditions over the next several decades

    Directory of Open Access Journals (Sweden)

    Masako Nakamura

    2012-02-01

    Ocean acidification is an ongoing threat for marine organisms due to the increasing atmospheric CO2 concentration. Seawater acidification has a serious impact on physiologic processes in marine organisms at all life stages. On the other hand, potential tolerance to external pH changes has been reported in coral larvae. Information about the possible mechanisms underlying such tolerance responses, however, is scarce. In the present study, we examined the effects of acidified seawater on the larvae of Acropora digitifera at the molecular level. We targeted two heat shock proteins, Hsp70 and Hsp90, and a heat shock transcription factor, Hsf1, because of their importance in stress responses and in early life developmental stages. Coral larvae were maintained under the ambient and elevated CO2 conditions that are expected to occur within next 100 years, and then we evaluated the expression of hsps and hsf1 by quantitative real-time polymerase chain reaction (PCR. Expression levels of these molecules significantly differed among target genes, but they did not change significantly between CO2 conditions. These findings indicate that the expression of hsps is not changed due to external pH changes, and suggest that tolerance to acidified seawater in coral larvae may not be related to hsp expression.

  16. Effect of Calmodulin (CaM) on Expression of Inducible Heat Shock Protein (HSP70) in Mice(Mus musculus) Preimplantation Embryo%钙调蛋白(CaM)对附植前小鼠胚胎热休克蛋白70(HSP70)表达的影响

    Institute of Scientific and Technical Information of China (English)

    索佳佳; 张保珍; 孙春玲; 王占美; 张双; 曹荣峰; 高善颂; 田文儒

    2013-01-01

    Heat shock protein(HSP)70 is synthesized in the preimplantation embryos of cow(Bos tarurs),ewe (Ovis aries) and mice(Mus musculus) under the conditions of heat shock in vitro,which enhances their thermotolerance to protect them from further heat damage.And Calmodulin(CaM)is the one of most important multifunctional receptor of Ca2 + and plays a key role in regulating the important gene expression in cell activities.To clarity whether the CaM participating in the expression of HSP70 in the mice embryos with heat shock and verify its mechanism,mice embryos were cultured in vitro to the blastocyst stage and then were randomly divided into control group(37℃),group treated with W7 without heat shock(37℃ +W7),heat shock group(39℃)and heat shock group treated with W7.The method of RT-PCR was used to detect the gene expression of CaM,HSP70 and heat shock factorl(HSF1),and the method of Western blot was used to detect the expression of CaM,HSP70 and HSF1.The combinations of both HSP70-CaM and HSP70-HSF1 were detected by using co-immunoprecipitation(Co-IP).The results showed that the CaM and HSP70 mRNA expression significantly increased in the mice embryos treated with 39℃ for 1 h (P<0.05).However,the CaM and HSP70 mRNA expression significantly decreased in both groups of embryo cultured at 37℃ (P<0.05) and 39℃ (P<0.01) treated with W7; W7 had no effect on the HSF1 mRNA expression in any groups of embryo.The synthesis of CaM,HSP70 and HSF1 significantly increased in the embryos treated at 39℃ for 1 h (P<0.05),and the CaM synthesis significantly decreased in the embryos cultured both at 37℃ and 39℃ and treated with W7 (P<0.05).W7 had no effect on neither HSP70 nor HSF1 synthesis in the embryos cultured at 37℃ (P>0.05) while it inhibited both HSP70 and HSF1 synthesis in the embryos cultured at 39℃ (P<0.05).The study also found that CaM combined with HSP70 and formed CaM-HSP70 complex in the mice embryos.The identification of interactions

  17. Determination of genomic diversity within mutant lemon (Citrus limon L.) and mandarin (Citrus reticulata) using molecular markers

    International Nuclear Information System (INIS)

    Mutation breeding is considered to be successful in order to obtain new cultivars and expanding the genetic source of new varieties of lemon and mandarin in Citrus. In this study, different doses of gamma radiation (60Co) have been used to generate novel mutants of the common variety Yerli Yuvarlak lemon and Yerli mandarin. The differences within the mutants were examined using simple sequence repeats (SSR) and sequence related amplified polymorphism (SRAP) markers. A total of 26 SSR primers and 28 SRAP primer combinations were screened for linkage to the different traits. Six SSR primers were showed polymorphism and closely linked to shoot tip color in lemon. Whereas, all of SRAP primer combinations were not produce any scorable polymorphic bands in both Yerli Yuvarlak lemon and Yerli mandarin. In this study, SSR and SRAP molecular markers were used together for first time to determination the genetic differences in citrus mutation breeding in the world. (author)

  18. 香蕉果皮2个HSP70基因片段的克隆及其在热激和冷藏过程中的表达%Cloning and expression analysis of two fragments of HSP70 cDNA in the peel of banana during storage at high and low temperature

    Institute of Scientific and Technical Information of China (English)

    贺立红; 陈建业; 王海蓝; 邝健飞; 陆旺金

    2012-01-01

    温度逆境胁迫可以诱导热激蛋白的表达,而热激处理能够减轻果实冷害。为了探讨温度逆境对香蕉热激蛋白基因表达的调控、辨析香蕉热激蛋白基因的表达与果实冷害的关系,采用同源序列法分离了香蕉果皮HStVO基因序列一根据已经报道的HStVO基因的氨基酸保守序列设计引物、以香蕉果皮总RNA为模板,用RT—PCR方法克隆香蕉的HSP70cDNA,Northern杂交分析该基因在不同贮藏温度下的表达特征。结果得到2个序列不同的HStVO基因片段,分别命名为Ma—HSP70-1和Ma—HSP70—2,Ma—HSP70-1和肘n—HSP70—2之间的碱基同源性为86.9%,氨基酸同源性为97.1%。Northern杂交的结果表明,38℃短期热激处理诱导了Ma-HSP70—2基因的表达,低温冷藏能使2个基因的表达增强。并初步认为Ma-HSP70可能与香蕉果实耐冷性有关。%Many evidences have indicated that temperature stress can induce heat shock protein expression and heat treatment can alleviate chilling injury of fruits. In the present study, two HSP70 fragments were cloned from banana peel using homology-based method in order to investigate their expressions regulated by temperature stress and the relationship between their expressions and chilling injury. Using PCR de- generate primers designed with reference to the conserved amino acid sequences of known HSP70, total RNA extracted from banana peel was used as templates and HSPTO eDNA fragments were amplified by RT-PCR in order to study their expression characteristics under different environments by Northern hy- bridization. Two different eDNA fragments, named Ma-HSP70-1 and Ma-HSP70-2, were cloned. The homology between two HSP70 is 86.9% at nueleotide acid sequence level and 97.1% at amino acid se- quence level. Using Ma-HSP70-1 and Ma-HSP70-2 as probes, Northern hybridization showed that short-time heat pretreatment (38 ℃ )only induced the expression of Ma-HSP70

  19. 运动对心肌、骨骼肌HSP70表达影响的研究进展%Progress of the Research on the Effect of Exercises on HSP70 Expression in Cardiac and Skeletal Muscles

    Institute of Scientific and Technical Information of China (English)

    曹桂霞; 白俊伟

    2010-01-01

    热休克蛋白的分子伴侣、抗氧化、抗损伤、协同免疫及抗细胞凋亡等多种功能,不仅生物界对它越来越关注,而且运动医学界的学者也对它产生了浓厚的兴趣.HSP70作为细胞应激蛋白,对不同强度与形式的运动,其反应和适应有很大差异,这为我们进一步认识运动规律,从而进行科学的体育锻炼和运动训练提供了新的思路和方法.

  20. Acetylcholinesterase (AChE) and heat shock proteins (Hsp70) of gypsy moth (Lymantria dispar L.) larvae in response to long-term fluoranthene exposure.

    Science.gov (United States)

    Mrdaković, Marija; Ilijin, Larisa; Vlahović, Milena; Matić, Dragana; Gavrilović, Anja; Mrkonja, Aleksandra; Perić-Mataruga, Vesna

    2016-09-01

    Polycyclic aromatic hydrocarbons (PAHs) may affect biochemical and physiological processes in living organisms, thus impairing fitness related traits and influencing their populations. This imposes the need for providing early-warning signals of pollution. Our study aimed to examine changes in the activity of acetylcholinesterase (AChE) and the concentration of heat shock proteins (Hsp70) in homogenates of brain tissues of fifth instar gypsy moth (Lymantria dispar L.) larvae, exposed to the ubiquitous PAH, fluoranthene, supplemented to the rearing diet. Significantly increased activity of AChE in larvae fed on the diets with high fluoranthene concentrations suggests the necessity for elucidation of the role of AChE in these insects when exposed to PAH pollution. Significant induction of Hsp70 in gypsy moth larvae reared on the diets containing low fluoranthene concentrations, indicate that changes in the level of Hsp70 might be useful as an indicator of pollution in this widespread forest species. PMID:27343862

  1. Expression of the C-terminal Polypeptide of Planarian Heat Shock Protein 70 and Preparation of Its Polyclonal Antibody%日本三角涡虫热休克蛋白70(DjHSP70)C-端多肽表达及其抗血清制备

    Institute of Scientific and Technical Information of China (English)

    马克学; 娄昊; 陈广文; 刘德增

    2011-01-01

    首先运用在线生物学软件对日本三角涡虫(Degusia japonica)热休克蛋白70(DjHSP70)氨基酸序列进行亲水区分析,发现该蛋白C-端含有较多亲水性氨基酸,然后以该段多肽序列为基础构建原核表达载体.采用PCR方法扩增450 bp cDNA片段,编码DjHSP70 C-端150个氨基酸多肽.将双酶切的cDNA片段与pET-28a载体连接后导入BL21受体菌,在IPTG诱导下表达出21 ku融合蛋白,分子量与预期相符.该融合蛋白采用Ni2+ -NTA agarose树脂进行纯化,纯化结果电泳检测后经灰度扫描分析显示纯度在95%以上.融合蛋白免疫新西兰大白兔获得高效价的抗血清,Western blot检测显示该抗血清不仅具有很强的特异性,而且还识别小鼠HSP70.此项工作为进一步研究淡水涡虫抗逆性奠定了基础.%In this paper, hydrophilic domain of DjHSP70 was predicted by using intemet biosoftware. The DjHSP70 C-terminal contains numerous hydrophilic amino acids. Based on this hydrophilic domain of DjHSP70, the prokaryotic expression vector was constructed. PCR method was used to amplify 450 bp cDNA fragment encoding DjHSP70 C-terminal 150 amino acid polypeptides. After digested by Hind Ⅲ/Xho Ⅰ , this eDNA fragment was ligated to pET-28a expression vector. Recombinant plasmid was transformed into E. coii BL21 and a 21 ku fusion protein was expressed after the induction with IPTG, which was in agreement with the expected molecular weight. This fusion protein was purified using Ni2+ -NTA agarose and detected by SDSPAGE electrophoresis. Grayscale scanning showed that the purity of the purified protein was over 95%. The fusion protein was used as an antigen to immunize New Zealand rabbits to prepare the polyclonal antibody. The results showed that this anti-serum was not only very specific to DjHSP70, but also recognized mouse HSP70. This work has laid the foundation for further investigating stress responses in freshwater planarians.

  2. Novel Improved Synthesis of HSP70 Inhibitor, Pifithrin-μ. In Vitro Synergy Quantification of Pifithrin-μ Combined with Pt Drugs in Prostate and Colorectal Cancer Cells.

    Science.gov (United States)

    McKeon, Aoife M; Egan, Alan; Chandanshive, Jay; McMahon, Helena; Griffith, Darren M

    2016-01-01

    We describe a novel improved approach to the synthesis of the important and well-known heat shock protein 70 inhibitor (HSP70), pifithrin-μ, with corresponding and previously unreported characterisation. The first example of a combination study comprising HSP70 inhibitor pifithrin-μ and cisplatin or oxaliplatin is reported. We have determined, using the Chou-Talalay method, (i) moderate synergistic and synergistic effects in co-treating PC-3 prostate cancer cells with pifithrin-μ and cisplatin and (ii) significant synergistic effects including strong synergism in cotreating HT29 colorectal cancer cells with oxaliplatin and pifithrin-μ. PMID:27455212

  3. Novel Improved Synthesis of HSP70 Inhibitor, Pifithrin-μ. In Vitro Synergy Quantification of Pifithrin-μ Combined with Pt Drugs in Prostate and Colorectal Cancer Cells

    Directory of Open Access Journals (Sweden)

    Aoife M. McKeon

    2016-07-01

    Full Text Available We describe a novel improved approach to the synthesis of the important and well-known heat shock protein 70 inhibitor (HSP70, pifithrin-μ, with corresponding and previously unreported characterisation. The first example of a combination study comprising HSP70 inhibitor pifithrin-μ and cisplatin or oxaliplatin is reported. We have determined, using the Chou-Talalay method, (i moderate synergistic and synergistic effects in co-treating PC-3 prostate cancer cells with pifithrin-μ and cisplatin and (ii significant synergistic effects including strong synergism in cotreating HT29 colorectal cancer cells with oxaliplatin and pifithrin-μ.

  4. Complex patterns of geographic variation in heat tolerance and Hsp70 expression levels in the common frog Rana temporaria

    DEFF Research Database (Denmark)

    Sørensen, Jesper Givskov; Pekkonen, Minna; Lindgren, Beatrice; Loeschcke, Volker; Laurila, Anssi; Merilä, Juha

    2009-01-01

    1. We tested for geographical variation in heat tolerance and Hsp70 expression levels of Rana temporaria tadpoles along a 1500 km long latitudinal gradient in Sweden.   2. Temperature tolerance of the hatchling tadpoles did not differ among populations, but they tolerated stressful hot temperatures...... stressful temperature (36 °C) the northern population showed highest tolerance. 4. The heat shock pre-treatment increased heat tolerance of the tadpoles. The possible adaptive role of Hsp70 for thermal tolerance and climatic adaptation in R. temporaria tadpoles is discussed....

  5. The expression of p53 and hsp70 proteins after treatment with Annona muricata Linn leaf for activating apoptotic and lead to homeostasis program of Raji cells

    OpenAIRE

    Okid Parama Astirin; Adi Prayitno; Anif Nur Artanti; Meutia Srikandi Fitria; Dyah Ayu Witianingsih; Dwimei Ayudewandari Pranatami; Suhartono Taat Putra

    2014-01-01

    Purpose: Organic extracts of plant Annonaceae enhances apoptosis in animal cells and get the drives to reach a new homeostasis. The incidence rate of nasopharyngeal cancer in Indonesia is quite high. Protein 53Kd (p53) play a role in apoptosis process, being heat shock protein 70 (hsp70) play a role in homeostasis. The aim of this research is to identify the apoptotic effects of Annona muricata Linn leaf toward Raji cells by observing the p53 and hsp70 expression.Methods: Apoptotic assay was ...

  6. HSF1/HSP70通路抑制c-Jun氨基末端激酶的活化保护UVA诱导的HaCaT细胞凋亡%Protection of HSF1/HSP70 pathway on UVA-induced HaCaT cells apoptosis via inhibiting the activation of c-Jun N-terminal kinase

    Institute of Scientific and Technical Information of China (English)

    王晓雯; 王春波; 李丙华; 韩彦弢

    2012-01-01

    目的 观察热休克转录因子1( HSF1)与热休克蛋白70( HSP70)对紫外线A(UVA)诱导HaCaT细胞凋亡的保护作用及其机制.方法 建立8mJ/cm2 UVA辐射损伤HaCaT细胞的病理模型.将细胞随机分为对照组、8mJ/cm2 UVA照射组、HSP70转录抑制剂组(50 μmol/L槲皮素).Honechst 33258荧光染色观察细胞凋亡;蛋白质印迹法检测UVA辐射HaCaT细胞后p-HSF1和HSP70蛋白的经时变化及UVA辐射后孵育6h JNK(c-Jun氨基末端激酶)、p-JNK的蛋白表达;Real-Time PCR检测HSP70 mRNA的表达.结果 UVA辐射后HaCaT细胞内p-HSF1、HSP70蛋白表达量均出现先增加后减少的时间依赖性趋势,其中p-HSF1于lh开始增加,3h达高峰,HSP70于6h达高峰,24h基本恢复原始水平;UVA辐射前预先加入HSP70转录抑制剂槲皮素能显著抑制HSP70 mRNA的表达,增加p-JNK的表达量,同时Honechst 33258荧光染色观察其与UVA辐射组比较凋亡率明显升高.结论 8mJ/cm2 UVA辐射HaCaT细胞在一定时间内可使HSF1活化致HSP70表达增加.HSFl/HSP70通路对UVA诱导的HaCaT细胞凋亡具有保护作用,其机制与HSP70大量表达后抑制JNK的活化有关.%Objective To investigate the protective effect of heat shock factorl ( HSF1) and heat shock protein70 ( HSP70) on ultraviolet A ( UVA ) -induced HaCaT cells apoptosis and its mechanism. Methods The apoptotic HaCaT cell model was induced by UVA irradiation (8mJ/cm ). The cells were randomly divided into three groups, including a control group, a model group (8mJ/cm UVA) and a HSP70 transcription inhibitor group (50 μmol/L quercetin). The morphologic alteration of apoptotic cells was investigated by using Hoechst 33258 fluorescent staining. Western blotting was used to investigate protein expression levels of phosphorylated HSF1 and HSP70 at different time points, as well as c-Jun N-terminal kinase ( JNK ) andphosphorylated JNK were investigated after incubating for 6 hours following UVA irradiation. HSP70 mRNA was

  7. HSP70在喉乳头状瘤和声带白斑组织中的表达及其临床意义%Expressions of HSP70 in laryngeal papilloma and vocal leukoplakia tissues and their clinical significances

    Institute of Scientific and Technical Information of China (English)

    杨田田; 傅仲鹰; 王苹; 杨立新

    2012-01-01

    目的:探讨热休克蛋白70 (HSP70)及其上游调控因子HSF1在喉乳头状瘤和声带白斑中的表达,阐明其在喉癌发生中的作用.方法:应用免疫组织化学法检测6例声带白斑(声带白斑组)、6例喉乳头状瘤(喉乳头状瘤组)和7例声带息肉(对照组)的组织标本中HSP70和HSF1蛋白的表达.结果:HSP70和HSF1蛋白在喉乳头状瘤、声带白斑中的表达高于声带息肉,差异有统计学意义(P<0.05或P<0.01);声带白斑在所有组织中表达最高.对声带息肉、喉乳头状瘤、声带白斑中HSP70蛋白表达量与HSF1表达的关系进行直线回归分析,结果显示,HSP70蛋白表达量与HSF1的变化呈现正的直线相关(r=0.867,P<0.01).结论:HSP70和HSF1在声带白斑中的高表达可能与喉癌的发生密切相关,且HSP70和HSF1可能成为喉癌早期诊断的指标.

  8. HDJC9, a novel human type C DnaJ/HSP40 member interacts with and cochaperones HSP70 through the J domain

    International Nuclear Information System (INIS)

    HSP40s are a subfamily of heat shock proteins (HSPs) and play important roles in regulation of cell proliferation, survival and apoptosis by serving as chaperones for HSP70s. Up to date hundreds of HSP40 proteins derived from various species ranging from Escherichia coli to homo sapiens have been identified. Here we report the cloning and characterization of a novel human type C DnaJ homologue, HDJC9, containing a typical N-terminal J domain. HDJC9 is upregulated at both mRNA and protein levels upon various stress and mitogenic stimulations. HDJC9 is mainly localized in cell nuclei under normal culture conditions while it is transported into cytoplasm and plasma membrane upon heat shock stress through a non-classical and lipid-dependent pathway. HDJC9 can interact with HSP70s and activate the ATPase activity of HSP70s, both of which are dependent on the J domain. Our data suggest that HDJC9 is a novel cochaperone for HSP70s

  9. Expressions of HSP 70 and NF-κB in the peripheral blood lymphocyte of chronic gastritis patients of different syndrome patterns

    Institute of Scientific and Technical Information of China (English)

    胡玲

    2012-01-01

    Objective To study the expressions of heat shock protein 70 (HSP 70) and nuclear factor-kappa B(NF-κB) in the peripheral blood lymphocyte of chronic gastritis (CG) patients of Pi-Wei hygropyrexia syndrome (PWHS) and Pi-qi deficiency syndrome(PQDS),and to explore their correlation with Helicobacter pylori(Hp) infection. Methods

  10. C-terminal phosphorylation of Hsp70 and Hsp90 regulates alternate binding to co-chaperones CHIP and HOP to determine cellular protein folding/degradation balances

    Czech Academy of Sciences Publication Activity Database

    Műller, P.; Růčková, E.; Halada, Petr; Coates, P.J.; Hrstka, R.; Lane, D. P.; Vojtěšek, B.

    -, June 2012 (2012), s. 10.1038. ISSN 0950-9232 Institutional support: RVO:61388971 Keywords : Hsp70 * Hsp90 * CHIP Subject RIV: CE - Biochemistry Impact factor: 7.357, year: 2012 http://www.readcube.com/articles/10.1038/onc.2012.314

  11. 15-Deoxy-Δ12,14-prostaglandin J2-induced down-regulation of endothelial nitric oxide synthase in association with HSP70 induction

    International Nuclear Information System (INIS)

    A natural ligand of peroxisome proliferator-activated receptor γ (PPARγ), 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2), decreases endothelial nitric oxide synthase (eNOS) expression by an unknown mechanism. Here we found that 15d-PGJ2-induced eNOS reduction is inversely associated with heat shock protein 70 (HSP70) induction in endothelial cells. Treatment of cells with 15d-PGJ2 decreased eNOS protein expression in a concentration- and time-dependent manner, but independently of PPARγ with no effect on mRNA levels. Although 15d-PGJ2 elicited endothelial apoptosis, inhibition of both pan-caspases and cathepsins failed to reverse reduction of eNOS protein. Interestingly, we observed that 15d-PGJ2 induced HSP70 in a dose-dependent manner. Immunoprecipitation and heat shock treatment demonstrated that eNOS reduction was strongly related to HSP70 induction. Cellular fractionation revealed that treatment with 15d-PGJ2 increased eNOS distribution 2.5-fold from soluble to insoluble fractions. These findings provide new insights into mechanisms whereby eNOS regulation by 15d-PGJ2 is related to HSP70 induction

  12. The Changes of HSP70 in Skeletal Muscle after an Exhaustive Eccentric Exercise and the Effects of Acupuncture%一次力竭离心运动后大鼠骨骼肌HSP70的变化及针刺对其影响

    Institute of Scientific and Technical Information of China (English)

    李俊平; 马延超; 张炜

    2011-01-01

    Purpose:The purpose of this study was to observe the changes of HSP70 expression in rat gastrocnemius muscle after an exhaustive eccentric exercise,and to explore the effects of acupuncture on HSP70 expression in skeletal muscle after exercise.Method:Male SD rats were randomly divided into four groups: control group,non-acupuncture group and acupuncture group after an exhaustive eccentric exercise.The rats of non-acupuncture and acupuncture group were carried out an exhaustive eccentric exercise.After the exercise,the rats of acupuncture group were handled acupuncture on the gastrocnemius vertically from the further side(the angle of the needle was 30 degree) and through the belly of gastrocnemius where the needle stayed for 5 minutes.Then the gastrocnemius was obtained from rats of different groups.The expression of HSP70 of gastrocnemius was detected by western blot and immunofluorescence histochemical method.Results:1)The expression of HSP70 of gastrocnemius in non-acupuncture group were increased markedly immediately,12 hours and 24 hours after exhaustive eccentric exercise(P0.05).2)The expression of HSP70 of gastrocnemius in acupuncture group were increased markedly only 12 hours after exhaustive eccentric exercise(P0.05).3)The expression of HSP70 of gastrocnemius in acupuncture group was significantly lower than non-acupuncture group immediately and 24 hours after exhaustive eccentric exercise(P0.05).4)The expression of HSP70 of gastrocnemius in non-acupuncture group mainly located in cytoplasm.The expression of HSP70 of gastrocnemius in acupuncture group mainly located in nucleus immediately after exhaustive eccentric exercise,and then transferred to cytoplasm gradually.Conclusions:1)Immediately after an exhaustive eccentric exercise,HSP70 expression of rat gastrocnemius were induced markedly,and then decreased gradually.2) HSP70 expression of rat gastrocnemius induced by an exhaustive eccentric exercise might be

  13. A comparative study of the timecourse of the expression of the thermo‑inducible HSP70 gene in clinical and environmental isolates of Aspergillus fumigatus.

    Science.gov (United States)

    Sharafi, Golnaz; Khosravi, Ali Reza; Vahedi, Ghasem; Yahyaraeyat, Ramak; Abbasi, Teimur

    2016-05-01

    The internal environment within animals or humans provides different conditions to invading saprophytic fungal pathogens, requiring the differential regulation of genes in comparison to environmental conditions. Understanding the mechanisms by which pathogens regulate genes within the host may be key in determining pathogen behavior within the host and may additionally facilitate further investigation into novel therapeutic agents. The heat shock protein (HSP)70 gene and its associated proteins have been frequently reported to be among the most highly expressed and dominant proteins present within various locations at physiological temperatures. The present study examined relative gene expression levels of the HSP70 gene in Aspergillus fumigatus isolates from both clinical and environmental origins, at a range of temperature points (20, 30, 37 and 42˚C) over five days, using reverse transcription‑quantitative polymerase chain reaction, comparing with a standard A. fumigatus strain incubated at 25˚C. The results indicated a differential gene expression pattern for the environmental and clinical isolates. During the five days, the HSP70 expression levels in the clinical samples were higher than in the environmental samples. However, the difference in the expression levels between the two groups at 42˚C was reduced. The mean HSP70 expression level over the five incubation days demonstrated a gradual and continual increasing trend by temperature elevation in both groups at 30, 37 and 42˚C, however, at 20˚C both groups demonstrated reduced expression. The temperature shift from 20 to 42˚C resulted in HSP70 induction and up to a 10‑ and 8.6‑fold change in HSP70 expression levels on the fifth day of incubation in the clinical and environmental groups, respectively. In conclusion, incubation at 37 and 42˚C resulted in the highest expression levels in both experimental groups, with these temperature points important for the induction of HSP70

  14. Role of HSP70 in the regulation of the testicular apoptosis in a seasonal breeding teleost Prochilodus argenteus from the São Francisco River, Brazil.

    Science.gov (United States)

    Domingos, Fabricio F T; Thomé, Ralph G; Martinelli, Patrícia M; Sato, Yoshimi; Bazzoli, Nilo; Rizzo, Elizete

    2013-04-01

    This study investigated the relationship among heat shock protein 70 (HSP70), proliferating cell nuclear antigen (PCNA), and testicular apoptosis during a breeding cycle of Prochilodus argenteus, a neotropical migratory characiform fish of importance in commercial fishery from the São Francisco River basin. A total of 48 (12 fish/sampling) adult males were caught using casting and drifting nets in four samplings from June 2008 to March 2009. Immunohistochemistry, Western blotting, terminal transferase-mediated dUTP nick-end labeling (TUNEL), enzyme-linked immunosorbent assay (ELISA), and caspase-3 colorimetric assay were assessed in different phases of spermatogenesis. Labeling for HSP70 occurred in spermatogonia (SPG(A) 18.0±1.5 and SPGB 27.9±1.0 in 100 mm(2), respectively) and Sertoli cells in all sampling periods, with higher values in June (resting period) while spermatocytes were labeled in September (maturation period) and December (ripe period). For PCNA, immunoreaction was predominant in spermatogonia in June and September, while primary spermatocytes were labeled mainly in December (18.7±2.0). TUNEL-positive reaction occurred throughout the sampling periods, and labeling was detected in the nucleus of germ cells in all developmental phases, except spermatozoa. By ELISA, total HSP70 in testis increased significantly from June to December, and decreased in March (regression period), P<0.05. Caspase-3 activity decreased from June to December and increased in March. Taken together, our results suggest that HSP70 may protect the germ cells from caspase-3-dependent apoptosis during testicular activity and, reduction of HSP70 and increase of apoptosis contribute for testicular remodeling after the breeding season in wild populations of P. argenteus in the São Francisco River. PMID:23362090

  15. Induction of specific humoral and cellular immune responses in a mouse model following gene fusion of HSP70C and Hantaan virus Gn and S0.7 in an adenoviral vector.

    Directory of Open Access Journals (Sweden)

    Linfeng Cheng

    Full Text Available Heat shock proteins (HSPs display adjuvant functions when given as fusion proteins to enhance vaccination efficiency. To evaluate enhanced potency of Hantaan virus (HTNV glycoprotein (GP and nucleocapsid protein (NP immunogenicity by heat shock protein 70 (HSP70, a recombinant adenovirus rAd-GnS0.7-pCAG-HSP70C expression vector was developed by genetically linking the HSP70 C-terminal gene (HSP70 359-610 aa, HSP70C to the Gn and 0.7 kb fragment of the NP (aa1-274-S0.7. C57BL/6 mice were immunized with these recombinant adenoviral vectors. A series of immunological assays determined the immunogenicity of the recombinant adenoviral vectors. The results showed that rAd-GnS0.7-pCAG-HSP70C induced a stronger humoral and cellular immune response than other recombinant adenoviruses (rAd-GnS0.7-pCAG and rAd-GnS0.7 and the HFRS vaccine control. Animal protection experiments showed that rAd-GnS0.7-pCAG-HSP70C was effective at protecting C57BL/6 mice from HTNV infection. The results of the immunological experiments showed that HSP70C lead to enhanced vaccine potency, and suggested significant potential in the development of genetically engineered vaccines against HTNV.

  16. Dexamethasone regulates CFTR expression in Calu-3 cells with the involvement of chaperones HSP70 and HSP90.

    Directory of Open Access Journals (Sweden)

    Luiz Felipe M Prota

    Full Text Available BACKGROUND: Dexamethasone is widely used for pulmonary exacerbation in patients with cystic fibrosis, however, not much is known about the effects of glucocorticoids on the wild-type cystic fibrosis channel transmembrane regulator (CFTR. Our aim was to determine the effects of dexamethasone treatment on wild-type CFTR expression. METHODS AND RESULTS: Dose-response (1 nM to 10 µM and time course (3 to 48 h curves were generated for dexamethasone for mRNA expression in Calu-3 cells using a real-time PCR. Within 24 h, dexamethasone (10 nM showed a 0.3-fold decrease in CFTR mRNA expression, and a 3.2-fold increase in αENaC mRNA expression compared with control groups. Dexamethasone (10 nM induced a 1.97-fold increase in the total protein of wild-type CFTR, confirmed by inhibition by mifepristone. To access surface protein expression, biotinylation followed by Western blotting showed that dexamethasone treatment led to a 2.35-fold increase in the amount of CFTR in the cell surface compared with the untreated control groups. Once protein translation was inhibited with cycloheximide, dexamethasone could not increase the amount of CFTR protein. Protein stability was assessed by inhibition of protein synthesis with cycloheximide (50 µg/ml at different times in cells treated with dexamethasone and in untreated cells. Dexamethasone did not alter the degradation of wild-type CFTR. Assessment of the B band of CFTR within 15 min of metabolic pulse labeling showed a 1.5-fold increase in CFTR protein after treatment with dexamethasone for 24 h. Chaperone 90 (HSP90 binding to CFTR increased 1.55-fold after treatment with dexamethasone for 24 h, whereas chaperone 70 (HSP70 binding decreased 0.30 fold in an immunoprecipitation assay. CONCLUSION: Mature wild-type CFTR protein is regulated by dexamethasone post transcription, involving cotranslational mechanisms with HSP90 and HSP70, which enhances maturation and expression of wild-type CFTR.

  17. Influence of atorvastatin on plasma HSP70 and HSF1 in patients with cute coronary syndrome%阿托伐他汀对急性冠脉综合征患者血浆中HSP70、HSF1的影响

    Institute of Scientific and Technical Information of China (English)

    王水侠; 刑坤

    2012-01-01

    目的:探讨阿托伐他汀对急性冠脉综合征(ACS)患者血浆中HSP70、HSF1的影响.方法:选取正常者39例,阿托伐他汀治疗组48例,酶联免疫吸附法(ELISA)测定样本血浆中HSP70、HSF1表达的变化.结果:(1)给药前,阿托伐他汀治疗组血浆中HSP70、HSF1表达明显高于对照组(P<0.05);(2)给药后,阿托伐他汀治疗组患者血浆中HSP70、HSF1表达较给药前升高明显(P<0.01).结论:阿托伐他汀可通过诱导HSP70表达增多发挥抗ACS作用.

  18. Caracterización in silico de las proteínas del choque térmico Hsp70 y Hsp90 deBemisia tabaci (Hemiptera: Aleyrodidae y su posible actividad adaptativa

    Directory of Open Access Journals (Sweden)

    Eneida Torres Cabra

    2014-06-01

    Full Text Available La mosca blanca, Bemisia tabaci (Hemiptera: Aleyrodidae es una de las plagas más destructivas e invasivas en el mundo, ataca una gran cantidad de cultivos. Se adapta fácilmente a plantas hospederas y a nuevas regiones geográficas, lo que sugiere el desarrollo de mecanismos de control a daños producidos por factores estresantes. Las proteínas Hsp se expresanen los organismos como mecanismo de defensa, actúan como chaperonas en el correcto ensamblaje de las proteínas. En este estudio se realizó una caracterizaciónin silico de las proteínas Hsp70 y Hsp90 de B. tabaci, secuencias obtenidas de NCBI. La determinaciónde los perfiles de hidrofobicidad, polaridad, accesibilidady flexibilidad se obtuvieron con “ProScale” de ExPASy, el perfil de antigenicidad con JaMBW. La secuencia aminoacídica se analizó con GOR IV y SOPMA y la composición de aminoácidos con ProtParam. Para analizar el peso molecular, índice deinestabilidad, índice alifático y gradiente hidropático,con GRAVY. La estructura terciaria se obtuvo con HHpred, y ESyPred3D. Para validar las estructuras 3D se utilizó Procheck, What_check y errat. Hsp70 y Hsp90 de B. tabaci presentan valores bajos de hidrofobicidady altos de polaridad, flexibilidad y accesibilidad, características que le permiten a las proteínas extender su capacidad como chaperonas. La Hsp70tiene una estructura secundaria compuesta por 41-45% alfa hélices, 30-43% coil y menos del 6% en hoja plegada y la Hsp90 por 52 y 53% hélices, 26-34% coily 6% hoja plegada. Las Hsp juegan un rol importante en los insectos debido a su tamaño y corto ciclo de vida, pues la temperatura influye en su distribución y abundancia.

  19. Insulin Promotes Survival of Amyloid-Beta Oligomers Neuroblastoma Damaged Cells via Caspase 9 Inhibition and Hsp70 Upregulation

    Directory of Open Access Journals (Sweden)

    M. Di Carlo

    2010-01-01

    Full Text Available Alzheimer's disease (AD and type 2 diabetes are connected in a way that is still not completely understood, but insulin resistance has been implicated as a risk factor for developing AD. Here we show an evidence that insulin is capable of reducing cytotoxicity induced by Amyloid-beta peptides (A-beta in its oligomeric form in a dose-dependent manner. By TUNEL and biochemical assays we demonstrate that the recovery of the cell viability is obtained by inhibition of intrinsic apoptotic program, triggered by A-beta and involving caspase 9 and 3 activation. A protective role of insulin on mitochondrial damage is also shown by using Mito-red vital dye. Furthermore, A-beta activates the stress inducible Hsp70 protein in LAN5 cells and an overexpression is detectable after the addition of insulin, suggesting that this major induction is the necessary condition to activate a cell survival program. Together, these results may provide opportunities for the design of preventive and therapeutic strategies against AD.

  20. RNAi knockdown of Hop (Hsp70/Hsp90 organising protein) decreases invasion via MMP-2 down regulation.

    LENUS (Irish Health Repository)

    Walsh, Naomi

    2011-07-28

    We previously identified Hop as over expressed in invasive pancreatic cancer cell lines and malignant tissues of pancreatic cancer patients, suggesting an important role for Hop in the biology of invasive pancreatic cancer. Hop is a co-chaperone protein that binds to both Hsp70\\/Hsp90. We hypothesised that by targeting Hop, signalling pathways modulating invasion and client protein stabilisation involving Hsp90-dependent complexes may be altered. In this study, we show that Hop knockdown by small interfering (si)RNA reduces the invasion of pancreatic cancer cells, resulting in decreased expression of the downstream target gene, matrix metalloproteinases-2 (MMP-2). Hop in conditioned media co-immunoprecipitates with MMP-2, implicating a possible extracellular function for Hop. Knockdown of Hop expression also reduced expression levels of Hsp90 client proteins, HER2, Bcr-Abl, c-MET and v-Src. Furthermore, Hop is strongly expressed in high grade PanINs compared to lower PanIN grades, displaying differential localisation in invasive ductal pancreatic cancer, indicating that the localisation of Hop is an important factor in pancreatic tumours. Our data suggests that the attenuation of Hop expression inactivates key signal transduction proteins which may decrease the invasiveness of pancreatic cancer cells possibly through the modulation of Hsp90 activity. Therefore, targeting Hop in pancreatic cancer may constitute a viable strategy for targeted cancer therapy.

  1. Cry1Ab treatment has no effects on viability of cultured porcine intestinal cells, but triggers Hsp70 expression.

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    Angelika Bondzio

    Full Text Available In vitro testing can contribute to reduce the risk that the use of genetically modified (GM crops and their proteins show unintended toxic effects. Here we introduce a porcine intestinal cell culture (IPEC-J2 as appropriate in vitro model and tested the possible toxic potential of Cry1Ab protein, commonly expressed in GM-maize. For comprehensive risk assessment we used WST-1 conversion and ATP content as metabolic markers for proliferation, lactate dehydrogenase release as indicator for cells with compromised membrane and transepithelial electrical resistance as parameter indicating membrane barrier function. The results were compared to the effects of valinomycin, a potassium ionophore, known to induce cytotoxic effects in most mammalian cell types. Whereas no toxicity was observed after Cry1Ab treatment, valinomycin induced a decrease in IPEC-J2 viability. This was confirmed by dynamic monitoring of cellular responses. Additionally, two dimensional differential in-gel electrophoresis was performed. Only three proteins were differentially expressed. The functions of these proteins were associated with responses to stress. The up-regulation of heat shock protein Hsp70 was verified by Western blotting as well as by enzyme-linked immunosorbent assay and may be related to a protective function. These findings suggest that the combination of in vitro testing and proteomic analysis may serve as a promising tool for mechanism based safety assessment.

  2. Correlation between HSP70 and c-F0S expression and apoptosis in rats undergolng transient focal cerebral ischemla and reperfusion

    Institute of Scientific and Technical Information of China (English)

    Chen Liynn; Li Yizhao; Hah Danchun

    2000-01-01

    OBJECTIVE To compare the induction of c-Fos and HSP70 and the presence of apoptosis and the influence of Ginkgo biloba extract (EGb761) in transient focal cerebral ischemic reperfusion rats.BACKGROUND Proto ancogene activation and induction of heat shock protein (HSP) occur in response to cerebral ischemia, but the correlation between these proteins and apoptosis remains uncertain. METHODS Healthy wistar rats were randomized to the normal control group(Group A, n=4), the Sham-operated control group(Group B, n=4), the ischemia and reperfusion group(Group C, n=24), the EGb761 pre-treated ischemia and reperfusion group(Group D, n=24). The rats of Group C and Group D were subjected to transient left middle cerebral artery occlusion (MCAO) as described by Zea longa for 1 hour. RESULTS Immunohistochemical analysis revealed no c-Fos or HSP70-immunoreactivity in Group A and Group B rats. However, in Group C rats, c-Fos was expressed in ipsilateral superficial cortical layers at 1 hour after reperfusion. At 6 hours, c-Fos immunoreactivities were increased in the ipsilateral cortex and were present in the contralateral cortex, while HSP70 were induced beginning in the ipsilateral neurons of MCA distribution. At 12 hours, the expression of c-Fos reached top in superficial cortical layers. At 24 hours HSP70 immunoreactivities reached top both in ipsilateral cortex and in ipsilateral striatum. At 3 days after recirculation, HSP70 expression decreased. c-Fos expression disappeared at day 7 and HSP70 expression only occured in endothelial cells. TUNEL staining showed that there was no cell apoptosis in Gronp A or in Group B. However, in Group C, TUNEL-positive neurons were observed in the border of the penumbra-like area that surrounds the ischemic core at 6 hours following reperfusion and then the number of TUNEL-positive cells reduced gradually. The changes in expression of HSP70 and c-Fos at different time points in Group D was in accord with in Group C, but the number of

  3. Survival and hsp70 gene expression in Plutella xylostella and its larval parasitoid Diadegma insulare varied between slowly ramping and abrupt extreme temperature regimes.

    Directory of Open Access Journals (Sweden)

    Md Habibullah Bahar

    Full Text Available BACKGROUND: In nature, insects have evolved behavioural and physiological adaptations to cope with short term exposure to extreme temperatures. Extreme heat events may increase as a result of climate change; this in turn will affect insect population dynamics. We examined the effect of abrupt and ecologically relevant gradual exposure to high temperatures on the survival and hsp70 gene expression in diamondback moth (DBM adults and the parasitoid Diadegma insulare, as well as in parasitized and non-parasitized DBM larvae. PRINCIPAL FINDINGS: Tolerance to high temperatures in DBM adults was higher than in D. insulare adults. There was no difference in the survival of DBM adults between abrupt and ramped increases from 25 to 38°C; however, at 40°C survival was higher when the temperature increased gradually. In contrast, more D. insulare adults survived when the temperature was ramped rather than shifted abruptly to both 38 and 40°C. There was no heat stress effect of up to 40°C on the survival of either parasitized or non-parasitized DBM larvae. In adults of both species, more hsp70 expression was observed when temperatures increased abruptly to 38°C compared to ramping. In contrast, at 40°C significantly more expression was found in insects exposed to the ramping rather than the abrupt regime. Hsp70 expression level was in agreement with adult survival data and appears to be a good indicator of stress levels. In parasitized and non-parasitized larvae, hsp70 expression was significantly higher after abrupt shifts compared to ramping at both temperatures. CONCLUSIONS/SIGNIFICANCE: Hsp70 gene expression was responsive to extreme temperatures in both DBM and D. insulare, which may underlie the ability of these insects to survive in extreme temperatures. Survival and hsp70 expression upon abrupt changes are distinctly different from those after ramping indicating that experimental protocol must be considered before extrapolating laboratory

  4. Recovery of ovary size, follicle cell apoptosis, and HSP70 expression in fish exposed to bleached pulp mill effluent

    Energy Technology Data Exchange (ETDEWEB)

    Janz, D. M.; Weber, L. P. [Oklahoma State Univ., Stillwater, OK (United States); McMaster, M. E.; Munkittrrick, K. R. [Environment Canada, Burlington, ON (Canada); Van Der Kraak, G. [Guelph Univ., Dept. of Zoology, ON (Canada)

    2001-03-01

    Apoptosis of granulosa cells that provide hormonal support for the oocyte is the normal mechanism by which atresia ( reduced ovarian size, decreased fecundity, delayed sexual maturation, alterations in plasma sex steroid levels, etc) occurs in mammals, birds and possibly fish. The objective of this study is to determine ovarian cell apoptosis, gonadosomatic index (GSI) and heat shock protein (HSP70) expression during the growth stage of ovarian development in white sucker fish in order to compare samples of fish collected upstream and downstream of a bleached kraft pulp mill in Ontario. Fish for the study were collected in two different years, before and after the pulp mill undertook a number of improvements to eliminate the release of process chemicals. Results showed a 3.4-fold increase in ovarian cell apoptosis in growing white sucker collected four km downstream of the bleached kraft pulp mill in 1996 (before the improvements) compared to fish collected from upstream sources. The elevated ovarian cell apoptosis was associated with significant reduction in gonadosomatic index in fish collected downstream. There were no differences in ovarian cell apoptosis or gonadosomatic index between fish collected upstream and four km downstream of the mill in September 1998 (after the improvements.) Based on the results, it may be concluded that chronic stimulation of ovarian cell apoptosis by certain components of bleached kraft pulp mill effluents represents an important cellular mechanism for reducing the size of ovaries and other related reproductive responses in female fish exposed to these effluents. Although the specific effluent components are not known, the improvements undertaken between 1996 and 1998 resulted in significant enough recovery of these responses to justify the belief in a cause-effect relationship. 32 refs., 1 tab., 2 figs.

  5. A platform to view huntingtin exon 1 aggregation flux in the cell reveals divergent influences from chaperones hsp40 and hsp70.

    Science.gov (United States)

    Ormsby, Angelique R; Ramdzan, Yasmin M; Mok, Yee-Foong; Jovanoski, Kristijan D; Hatters, Danny M

    2013-12-27

    Our capacity for tracking how misfolded proteins aggregate inside a cell and how different aggregation states impact cell biology remains enigmatic. To address this, we built a new toolkit that enabled the high throughput tracking of individual cells enriched with polyglutamine-expanded Htt exon 1 (Httex1) monomers, oligomers, and inclusions using biosensors of aggregation state and flow cytometry pulse shape analysis. Supplemented with gel filtration chromatography and fluorescence-adapted sedimentation velocity analysis of cell lysates, we collated a multidimensional view of Httex1 aggregation in cells with respect to time, polyglutamine length, expression levels, cell survival, and overexpression of protein quality control chaperones hsp40 (DNAJB1) and hsp70 (HSPA1A). Cell death rates trended higher for Neuro2a cells containing Httex1 in inclusions than with Httex1 dispersed through the cytosol at time points of expression over 2 days. hsp40 stabilized monomers and suppressed inclusion formation but did not otherwise change Httex1 toxicity. hsp70, however, had no major effect on aggregation of Httex1 but increased the survival rate of cells with inclusions. hsp40 and hsp70 also increased levels of a second bicistronic reporter of Httex1 expression, mKate2, and increased total numbers of cells in culture, suggesting these chaperones partly rectify Httex1-induced deficiencies in quality control and growth rates. Collectively, these data suggest that Httex1 overstretches the protein quality control resources and that the defects can be partly rescued by overexpression of hsp40 and hsp70. Importantly, these effects occurred in a pronounced manner for soluble Httex1, which points to Httex1 aggregation occurring subsequently to more acute impacts on the cell. PMID:24196953

  6. A Platform to View Huntingtin Exon 1 Aggregation Flux in the Cell Reveals Divergent Influences from Chaperones hsp40 and hsp70*

    Science.gov (United States)

    Ormsby, Angelique R.; Ramdzan, Yasmin M.; Mok, Yee-Foong; Jovanoski, Kristijan D.; Hatters, Danny M.

    2013-01-01

    Our capacity for tracking how misfolded proteins aggregate inside a cell and how different aggregation states impact cell biology remains enigmatic. To address this, we built a new toolkit that enabled the high throughput tracking of individual cells enriched with polyglutamine-expanded Htt exon 1 (Httex1) monomers, oligomers, and inclusions using biosensors of aggregation state and flow cytometry pulse shape analysis. Supplemented with gel filtration chromatography and fluorescence-adapted sedimentation velocity analysis of cell lysates, we collated a multidimensional view of Httex1 aggregation in cells with respect to time, polyglutamine length, expression levels, cell survival, and overexpression of protein quality control chaperones hsp40 (DNAJB1) and hsp70 (HSPA1A). Cell death rates trended higher for Neuro2a cells containing Httex1 in inclusions than with Httex1 dispersed through the cytosol at time points of expression over 2 days. hsp40 stabilized monomers and suppressed inclusion formation but did not otherwise change Httex1 toxicity. hsp70, however, had no major effect on aggregation of Httex1 but increased the survival rate of cells with inclusions. hsp40 and hsp70 also increased levels of a second bicistronic reporter of Httex1 expression, mKate2, and increased total numbers of cells in culture, suggesting these chaperones partly rectify Httex1-induced deficiencies in quality control and growth rates. Collectively, these data suggest that Httex1 overstretches the protein quality control resources and that the defects can be partly rescued by overexpression of hsp40 and hsp70. Importantly, these effects occurred in a pronounced manner for soluble Httex1, which points to Httex1 aggregation occurring subsequently to more acute impacts on the cell. PMID:24196953

  7. Generation of the First TCR Transgenic Mouse with CD4+ T Cells Recognizing an Anti-inflammatory Regulatory T Cell-Inducing Hsp70 Peptide

    Science.gov (United States)

    Jansen, Manon A. A.; van Herwijnen, Martijn J. C.; van Kooten, Peter J. S.; Hoek, Aad; van der Zee, Ruurd; van Eden, Willem; Broere, Femke

    2016-01-01

    Antigen-specific regulatory T cells (Tregs) directed at self-antigens are difficult to study since suitable specific tools to isolate and characterize these cells are lacking. A T cell receptor (TCR)-transgenic mouse would generate possibilities to study such ­antigen-specific T cells. As was shown previously, immunization with the mycobacterial heat shock protein (Hsp) 70-derived peptide B29 and its mouse homologs mB29a and mB29b induced anti-inflammatory responses. Furthermore, B29 induced antigen-­specific Tregs in vivo. To study mB29b-specific Tregs, we isolated the TCR from T cell hybridomas generated against mB29b and produced a TCR transgenic mouse that expresses a MHC-class II restricted mB29b-specific TCR. These TCR transgenic CD4+ T cells were found to cross-react with the B29 epitope as identified with peptide-induced proliferation and IL-2 production. Thus, we have successfully generated a novel mouse model with antigen-specific CD4+ T cells that recognize self and bacterial Hsp 70-derived peptides. With this novel mouse model, it will be possible to study primary antigen-specific T cells with specificity for a regulatory Hsp70 T cell epitope. This will enable the isolation and characterization CD4+CD25+ Tregs with a proven specificity. This will provide useful knowledge of the induction, activation, and mode of action of Hsp70-specific Tregs, for instance, during experimental arthritis. PMID:27014269

  8. Generation of the First TCR Transgenic Mouse with CD4(+) T Cells Recognizing an Anti-inflammatory Regulatory T Cell-Inducing Hsp70 Peptide.

    Science.gov (United States)

    Jansen, Manon A A; van Herwijnen, Martijn J C; van Kooten, Peter J S; Hoek, Aad; van der Zee, Ruurd; van Eden, Willem; Broere, Femke

    2016-01-01

    Antigen-specific regulatory T cells (Tregs) directed at self-antigens are difficult to study since suitable specific tools to isolate and characterize these cells are lacking. A T cell receptor (TCR)-transgenic mouse would generate possibilities to study such -antigen-specific T cells. As was shown previously, immunization with the mycobacterial heat shock protein (Hsp) 70-derived peptide B29 and its mouse homologs mB29a and mB29b induced anti-inflammatory responses. Furthermore, B29 induced antigen--specific Tregs in vivo. To study mB29b-specific Tregs, we isolated the TCR from T cell hybridomas generated against mB29b and produced a TCR transgenic mouse that expresses a MHC-class II restricted mB29b-specific TCR. These TCR transgenic CD4(+) T cells were found to cross-react with the B29 epitope as identified with peptide-induced proliferation and IL-2 production. Thus, we have successfully generated a novel mouse model with antigen-specific CD4(+) T cells that recognize self and bacterial Hsp 70-derived peptides. With this novel mouse model, it will be possible to study primary antigen-specific T cells with specificity for a regulatory Hsp70 T cell epitope. This will enable the isolation and characterization CD4(+)CD25(+) Tregs with a proven specificity. This will provide useful knowledge of the induction, activation, and mode of action of Hsp70-specific Tregs, for instance, during experimental arthritis. PMID:27014269

  9. Effects of heavy metals on ultrastructure and Hsp70 induction in Lemna minor L. exposed to water along the Sarno River, Italy.

    Science.gov (United States)

    Basile, A; Sorbo, S; Cardi, M; Lentini, M; Castiglia, D; Cianciullo, P; Conte, B; Loppi, S; Esposito, S

    2015-04-01

    The effects of freshwater pollution in the highly contaminated river Sarno (Campania, Southern Italy) have been evaluated using bags containing the aquatic plant Lemna minor (Lemnacee, Arales), in order to determine morpho-physiological modifications as a response to pollutants. The exposition of Lemna bags for 7 days on three different sites along the river path showed alterations in chloroplasts and vacuoles shape and organization. Moreover, some specimens were exposed in vitro at the same heavy metal (HM) concentrations measured in the polluted sites of the river, and compared with data from the bag experiment; to verify the dose and time dependent effects, samples were exposed to HM in vitro at concentrations ranging from 10(-6) to 10(-4)M up to 7 days. Transmission electron microscopy (TEM) observations on in vitro plants confirmed that ultrastructural alterations affected most of plastids and the shape of different subcellular structures, namely vacuoles; in in vitro stressed specimens, Heat Shock Proteins 70 (Hsp70) levels changed, in dependence of changing levels of HM measured in different sites along the river path. Thus L. minor exhibited a possible correlation between the levels of HM pollution and Hsp70 occurrence; interestingly, the data presented showed that copper specifically increased Hsp70 levels at concentrations detected in polluted river waters, whereas cadmium and lead did not; on the other side, the latter represent highly toxic elements when specimens were exposed to higher levels in vitro. The effects of specific elements in vitro are compared to those observed in bags exposed along the river path; thus results are examined in order to propose L. minor as an organism able to be utilized to monitor heavy metals pollution; the possibility of using Hsp70s as specific markers of HM pollution is discussed. PMID:25618632

  10. A Novel Strategy for TNF-Alpha Production by 2-APB Induced Downregulated SOCE and Upregulated HSP70 in O. tsutsugamushi-Infected Human Macrophages

    Science.gov (United States)

    Liang, Jui-Lin; Tsai, Ming-Hsien; Yen, Chia-Jung; Li, Hsiu-Wen; Chiu, Siou-Jin; Chang, Chung-Hsing; Huang, Yaw-Bin; Lin, Ming-Wei; Yoshioka, Tohru

    2016-01-01

    Orientia (O.) tsutsugamushi-induced scrub typhus is endemic across many regions of Asia and the Western Pacific, where an estimated 1 million cases occur each year; the majority of patients infected with O. tsutsugamushi end up with a cytokine storm from a severe inflammatory response. Previous reports have indicated that blocking tumor necrosis factor (TNF)-α reduced cell injury from a cytokine storm. Since TNF-α production is known to be associated with intracellular Ca2+ elevation, we examined the effect of store-operated Ca2+ entry (SOCE) inhibitors on TNF-α production in O. tsutsugamushi-infected macrophages. We found that 2-aminoethoxydiphenyl borate (2-APB), but not SKF96365, facilitates the suppression of Ca2+ mobilization via the interruption of Orai1 expression in O. tsutsugamushi-infected macrophages. Due to the decrease of Ca2+ elevation, the expression of TNF-α and its release from macrophages was repressed by 2-APB. In addition, a novel role of 2-APB was found in macrophages that causes the upregulation of heat shock protein 70 (HSP70) expression associated with ERK activation; upregulated TNF-α production in the case of knockdown HSP70 was inhibited with 2-APB treatment. Furthermore, elevated HSP70 formation unexpectedly did not help the cell survival of O. tsutsugamushi-infected macrophages. In conclusion, the parallelism between downregulated Ca2+ mobilization via SOCE and upregulated HSP70 after treatment with 2-APB against TNF-α production was found to efficiently attenuate an O. tsutsugamushi-induced severe inflammatory response. PMID:27472555

  11. Caries Experience among Females aged 16–21 in Punjab, India and its Relationship with Lifestyle and Salivary HSP70 Levels

    OpenAIRE

    Kaur, Rabinder; Kataria, Hardeep; Kumar, Sushil; Kaur, Gurcharan

    2010-01-01

    Objectives: The objective of the current study was to assess the prevalence and means of dental caries experience and the prevalence of calculus among college students and their awareness of general dental health. The correlation between the experience of caries and extracellular levels of heat shock protein-70 (eHSP70) was studied in saliva samples of these students to ascertain whether eHSP level changes as a result of increased caries experience of these subjects. Methods: The current stud...

  12. Analyses of promoter-proximal pausing by RNA polymerase II on the hsp70 heat shock gene promoter in a Drosophila nuclear extract.

    OpenAIRE

    Li, B.; Weber, J. A.; Chen, Y; Greenleaf, A L; Gilmour, D S

    1996-01-01

    Analyses of Drosophila cells have revealed that RNA polymerase II is paused in a region 20 to 40 nucleotides downstream from the transcription start site of the hsp70 heat shock gene when the gene is not transcriptionally active. We have developed a cell-free system that reconstitutes this promoter-proximal pausing. The paused polymerase has been detected by monitoring the hyperreactivity of thymines in the transcription bubble toward potassium permanganate. The pattern of permanganate reacti...

  13. Allele-specific PCR genotyping of the HSP70 gene polymorphism discriminating the green and red color variants sea cucumber (Apostichopus japonicus)

    Institute of Scientific and Technical Information of China (English)

    Jung-Ha Kang; Ki Hwan Yu; Jung-Youn Park; Chul-Min An; Je-Cheon Jun; Sang-Jun Lee

    2011-01-01

    Color variation is a well-known feature of sea cucumbers (Apostichopus japonicus),which are classified into three groups based on their colors of red,green and black.It is also one of the most important traits related to how they taste,and it thereby affects their market price.Attempts were made to identify single-nucleotide polymorphisms (SNPs) and to analyze differences associated with SNP genotypes between green and red color variants using HSP70 as the target gene.The HSP70 gene,which is found universally in organisms from bacteria to humans,is one of the most evolutionarily conserved genes and the most widely studied biomarker of stress response.DNA fragments of 1074 bp covering a partial sequence of the sea cucumber HSP70 gene,were amplified from both red and green variants,and subsequently analyzed for the presence of SNPs.Twenty-seven polymorphic sites in total,including heterozygous sites,were observed.Of these,six sites were found to be significantly different SNP genotypes between green and red variants.Furthermore,PCR with an internal primer designed to include an allelespecific SNP at the 3' end (site 443) showed differentiation between the two variants,100% and 4.2% amplification in green and red variants,respectively.The validated SNPs may serve as informative genetic markers that can be used to distinguish variants at the early developmental stage,prior to color differentiation.

  14. Ekspresi Human Leukocyte Antigen-G (HLA-G dan Heat-Shock Protein-70 (Hsp-70 pada Pertumbuhan Janin Terhambat

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    Sri Sulistyowati

    2014-03-01

    Full Text Available Intra uterine growth retardation (IUGR is one of the leading causes of higher morbidity and mortality in perinatal. Immune maladaptation affects trophoblast invasion and spiralis arteria remodeling that will cause placental tissue hypoxia. This research aimed to analyze human leukocyte antigen-G (HLA-G and heat-shock protein-70 (Hsp-70 expression on the IUGR trophoblast and normal pregnancy, by applying analytical observational method and cross sectional approach. This research was conducted at the Obstetric and Gynecology Department of Dr. Moewardi Hospital Surakarta from November to December 2011. The total samples were 30, divided into two groups. There were 15 samples trophoblast on IUGR and 15 samples trophoblast from normal pregnancy. All samples were tested for HLA-G and Hsp-70 using immunohistochemistry. The data were analyzed by using t-test. The mean of HLA-G expression on the IUGR groups was 32.42±8.90 and on the normal pregnancy groups was 43.92±14.91 (p=0.016. Heat-shock protein70 expression on the IUGR groups was 2.4355+0.26647 and on the normal pregnancy groups was 1.5920+0.17142 with p=0.008. In conclusion, in IUGR, the HLA-G expression is lower and the Hsp-70 expression is higher than in normal pregnancy.

  15. Induction of heat shock proteins (hsp70) in the zebra mussel (Dreissena polymorpha) following exposure to platinum group metals (platinum, palladium and rhodium): Comparison with lead and cadmium exposures

    International Nuclear Information System (INIS)

    An increasing number of papers concentrate on the availability and uptake of platinum group elements (PGE) by different organisms. These metals are discharged into the environment from different anthropogenic sources, such as automobile catalytic converters, hospitals and other medical institutions. As the effects of these precious metals on organisms remain unclear, the induction of heat shock proteins (hsp70) in zebra mussels (Dreissena polymorpha) following exposure to soluble salts of platinum, palladium and rhodium was compared with the hsp70 induction in mussels following exposure to cadmium and lead. Mussels were sampled weekly during a period of 10 weeks and analyzed for their metal concentration and their hsp70 level. Highest metal uptake was found for Cd, followed by Pt, Pb and Pd. Rh demonstrated the lowest uptake rate. A clear time-dependent increase of hsp70 levels occurred in all exposed mussels. Concentrations of hsp70 started to rise between days 18 and 25, except for the Pt-exposed group, where the initial increase was between days 25 and 32. All groups reached maximal hsp70 concentrations at day 39. Subsequently, hsp70 levels decreased to initial levels for the remaining exposure period. Threshold metal levels for the hsp70 induction varied among the metals and increased in the order: Rh < Pd ≤ Pb < Pt < Cd. Highest hsp70 values were found for mussels exposed to Pd, with a 25-fold higher level than in the controls, followed by Pt- and Rh-exposed mussels, which showed a 19-fold increase. The hsp70 levels of the mussels exposed to Cd and Pb were much lower, showing 6- and 12-fold higher values than the control, respectively. The clear induction of hsp70 due to exposure to Pt, Pd and Rh gives evidence for strong cellular effects of these metals, especially, when compared with lead and cadmium. Among the metals tested, Pd seems to have the highest potential as inducer for hsp70 production due to its low threshold level in combination with the

  16. The specialized Hsp70 (HscA) interdomain linker binds to its nucleotide-binding domain and stimulates ATP hydrolysis in both cis and trans configurations.

    Science.gov (United States)

    Alderson, T Reid; Kim, Jin Hae; Cai, Kai; Frederick, Ronnie O; Tonelli, Marco; Markley, John L

    2014-11-25

    Proteins from the isc operon of Escherichia coli constitute the machinery used to synthesize iron-sulfur (Fe-S) clusters for delivery to recipient apoproteins. Efficient and rapid [2Fe-2S] cluster transfer from the holo-scaffold protein IscU depends on ATP hydrolysis in the nucleotide-binding domain (NBD) of HscA, a specialized Hsp70-type molecular chaperone with low intrinsic ATPase activity (0.02 min(-1) at 25 °C, henceforth reported in units of min(-1)). HscB, an Hsp40-type cochaperone, binds to HscA and stimulates ATP hydrolysis to promote cluster transfer, yet while the interactions between HscA and HscB have been investigated, the role of HscA's interdomain linker in modulating ATPase activity has not been explored. To address this issue, we created three variants of the 40 kDa NBD of HscA: NBD alone (HscA386), NBD with a partial linker (HscA389), and NBD with the full linker (HscA395). We found that the rate of ATP hydrolysis of HscA395 (0.45 min(-1)) is nearly 15-fold higher than that of HscA386 (0.035 min(-1)), although their apparent affinities for ATP are equivalent. HscA395, which contains the full covalently linked linker peptide, exhibited intrinsic tryptophan fluorescence emission and basal thermostability that were higher than those of HscA386. Furthermore, HscA395 displayed narrower (1)H(N) line widths in its two-dimensional (1)H-(15)N TROSY-HSQC spectrum in comparison to HscA386, indicating that the peptide in the cis configuration binds to and stabilizes the structure of the NBD. The addition to HscA386 of a synthetic peptide with a sequence identical to that of the interdomain linker (L(387)LLDVIPLS(395)) stimulated its ATPase activity and induced widespread NMR chemical shift perturbations indicative of a binding interaction in the trans configuration. PMID:25372495

  17. Gastric Ulcers in Middle-Aged Rats: The Healing Effect of Essential Oil from Citrus aurantium L. (Rutaceae

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    C. M. Polo

    2012-01-01

    Full Text Available The elderly population has experienced increased life expectancy as well as the increased incidence of gastric ulcers. The peels of fruits from Citrus aurantium L., popularly known in Brazil as orange bitter, are commonly used asatea form for the treatment of gastrointestinal tract disorders, such as ulcer and gastritis. We evaluated the healing effects of essential oil from the peels of Citrus aurantium fruits (OEC on gastric ulcers in middle-aged rats. We examined the effects of a 14-day chronic OEC treatment on gastric mucosa in middle-aged male Wistar rats that were given acetic-acid-induced gastric lesions by morphometric and immunohistological analyses. Oral OEC treatment significantly reduced the lesion area (76% within the gastric mucosa and significantly increased (P<.05 the height of regenerated mucosa (59% when compared to the negative control group. Immunohistochemical analysis of the molecular markers such as COX-2, HSP-70, VEGF, and PCNA in the gastric mucosa confirmed that OEC treatment induced healing effects by increasing the number of new blood vessels and by augmenting gastric mucus in the mucosa glands. These results suggest that the oil from Citrus aurantium effectively heals gastric ulcers in middle-aged animals; however, safe use of OEC demands special care and precautions.

  18. 钙/钙调素依赖性蛋白激酶Ⅱ对热休克小鼠胚胎成纤维细胞HSP70表达的影响%Calcium/Calmodulin-dependent Protein Kinase Ⅱ Contributes to HSP70 Expression in Mouse Embryonic Fibroblasts

    Institute of Scientific and Technical Information of China (English)

    孙琪; 丛霞; 索佳佳; 曹荣峰; 姜忠玲; 崔凯; 高善颂; 田文儒

    2012-01-01

    The objectives of the study were to verify if the CaMK II (Ca +/calmodulin-dependent protein kinase II ) influences the expression of HSP70 gene in the mouse embryonic fibroblasts (MEF) with heat shock and to clarify its mechanism. The fibroblasts were heat-treated at 37t ,39^ and 41°C individually before the expressions of both CaMK II and HSF1 were detected by RT-PCR at 0. 5 ,1 ,1. 5 and 2 h of heat shock ,respectivelly. Moreover, the fibroblasts were divided into control group and myr-AIP group randomly,and cultured at 371 and 39°C respectively. The expressions of both HSP70 and HSF1 were detected. The expressions of HSF1 and CaMK II mRNA in the MEF treated at 39^ for 1 h were both extremely significantly higher (P <0. 01) than that of the control group. There was a significantly (P <0. 05) decrease of HSP70 expression in the MEF cultured at 39t treated with myr-AIP compared with its blank control. However, there was no difference in HSP70 exoression between the MEF cultured at 37X1 with or without myr-AIP. The expressions of HSF1 in the MEF treated with myr-AIP were not significantly influenced. However,the p-HSFl expression in the MEF treated with myr-AIP was significantly (P <0. 05) decreased. Moderate heat shock increases both expressions of CaMK FJ and HSF1 in mouse embryonic fibroblasts. CaMK II participates in both heat shock response and HSP70 gene expression by p-HSFl.%为证实热休克小鼠胎儿成纤维细胞中钙/钙调素依赖性蛋白激酶Ⅱ(Ca2+/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)对热休克蛋白70 (Heat shock protein70,HSP70)基因表达的影响及其作用机理,将体外培养的小鼠胎儿成纤维细胞(Mouse Embryonic Fibroblasts,MEF)随机分为39℃和41℃热处理组(分别处理0.5,1,1.5,2h)和常温对照组(37℃),测定各组细胞CaMKⅡ和HSF1 mRNA的量.此外,将培养的细胞分为37℃和39℃CaMKⅡ特异性抑制剂(myr-AIP)处理组和相应的空白对照组,分别测定各组细胞HSP70、HSF1

  19. Specific anti-tumor immune responses of dendritic cells pulsed with recombinant human rhHSP70 and freeze-thaw cellular lysates derived from breast cancer%rhHSP70联合冻融抗原修饰树突状细胞诱导的抗乳腺癌作用*

    Institute of Scientific and Technical Information of China (English)

    李斌; 陈鹏; 郑建云

    2013-01-01

      目的:利用rhHSP70联合树突状细胞递呈肿瘤抗原的特性提高细胞毒T淋巴细胞(CTLs)对乳腺癌细胞的杀伤活性。方法:外周血单个核细胞体外经GM-CSF和IL-4诱导产生树突状细胞,负载冻融抗原肽的同时加入新型热休克蛋白(rhHSP70),不同分组分别诱导自体CTLs产生。ELISA测定CTLs杀伤活性和细胞因子的分泌。结果:冻融抗原肽致敏的DCs促进CTLs增殖,上调CTLs中CD3+和CD8+T细胞群及Th1型细胞因子的分泌;体外实验中具有对人乳腺癌细胞MCF-7的杀伤活性,在加入rhHSP70后效果更加明显,并能显著增强CTLs对肿瘤细胞的杀伤率。结论:rhHSP70联合肝癌冻融抗原修饰DCs,能够促进DCs的成熟,增强DCs刺激淋巴细胞增殖的能力,诱导的CTLs在体外对乳腺癌细胞能产生高效杀伤力。rhHSP70增强DCs抗肿瘤能力的机制可能与其促进DCs成熟有关。%Objective:This work aims to use the characteristics of dendritic cells (DCs) pulsed with recombinant human HSP70, which can present and process tumor antigens, to enhance the killing activity of cytotoxic t lymphocytes (CTLs) against breast neoplasms. Methods:Autologous DCs were isolated from peripheral blood mononuclear cells and then stimulated in vitro with granulocyte macrophage-colony stimulating factor and interleukin-4. The DCs were loaded with A549 tumor cell freeze-thaw lysate, and rhHSP70 was added as an immune adjuvant. The specific groups were subjected to tumor-specific cytotoxic assay, enzyme-linked immunosorbent assay, and fluores-cence-activated cell sorting. Results:DCs pulsed with A549 tumor cell lysate enhanced the growth expansion of CTLs, upregulated CD40 and CD80 populations in CTLs, and augmented Th1 cytokines. In addition, the cytotoxicity of specific CTLs against A549 was highly enhanced. The above indications became more obvious after the addition of rhHSP70. Conclusion:DCs pulsed with freeze-thaw cell

  20. Phylogenetic and Molecular Variability Studies Reveal a New Genetic Clade of Citrus leprosis virus C

    Science.gov (United States)

    Ramos-González, Pedro Luis; Chabi-Jesus, Camila; Guerra-Peraza, Orlene; Breton, Michèle Claire; Arena, Gabriella Dias; Nunes, Maria Andreia; Kitajima, Elliot Watanabe; Machado, Marcos Antonio; Freitas-Astúa, Juliana

    2016-01-01

    Citrus leprosis virus C (CiLV-C) causes a severe disease affecting citrus orchards in the Western hemisphere. This study reveals the molecular variability of the virus by analyzing four genomic regions (p29, p15, MP and RNA2-intergenic region) distributed over its two RNAs. Nucleotide diversity (π) values were relatively low but statistically different over the analyzed genes and subpopulations, indicating their distinct evolutionary history. Values of πp29 and πMP were higher than those of πp15 and πRNA2–IR, whereas πMP was increased due to novel discovered isolates phylogenetically clustered in a divergent clade that we called SJP. Isolate BR_SP_SJP_01 RNA1 and RNA2 sequences, clade SJP, showed an identity of 85.6% and 88.4%, respectively, with those corresponding to CiLV-C, the type member of the genus Cilevirus, and its RNA2 5′-proximal region was revealed as a minor donor in a putative inter-clade recombination event. In addition to citrus, BR_SP_SJP_01 naturally infects the weed Commelina benghalensis and is efficiently transmitted by Brevipalpus yothersi mites. Our data demonstrated that negative selection was the major force operating in the evaluated viral coding regions and defined amino acids putatively relevant for the biological function of cilevirus proteins. This work provides molecular tools and sets up a framework for further epidemiological studies. PMID:27275832

  1. Divergent ecological histories of two sister Antarctic krill species led to contrasted patterns of genetic diversity in their heat-shock protein (hsp70) arsenal.

    Science.gov (United States)

    Papot, Claire; Cascella, Kévin; Toullec, Jean-Yves; Jollivet, Didier

    2016-03-01

    The Arctic and the Antarctic Peninsula are currently experiencing some of the most rapid rates of ocean warming on the planet. This raises the question of how the initial adaptation to extreme cold temperatures was put in place and whether or not directional selection has led to the loss of genetic variation at key adaptive systems, and thus polar species' (re)adaptability to higher temperatures. In the Southern Ocean, krill represents the most abundant fauna and is a critical member at the base of the Antarctic food web. To better understand the role of selection in shaping current patterns of polymorphisms, we examined genetic diversity of the cox-1 and hsp70 genes by comparing two closely related species of Euphausiid that differ in ecology. Results on mtcox-1 agreed with previous studies, indicating high and similar effective population sizes. However, a coalescent-based approach on hsp70 genes highlighted the role of positive selection and past demographic changes in their recent evolution. Firstly, some form of balancing selection was acting on the inducible isoform C, which reflected the maintenance of an ancestral adaptive polymorphism in both species. Secondly, E. crystallorophias seems to have lost most of its hsp70 diversity because of a population crash and/or directional selection to cold. Nonsynonymous diversities were always greater in E. superba, suggesting that it might have evolved under more heterogeneous conditions. This can be linked to species' ecology with E. superba living in more variable pelagic conditions, while E. crystallorophias is strictly associated with continental shelves and sea ice. PMID:27087928

  2. 热应激预处理影响鼠背轴型皮瓣HSP70、HSF1表达的实验研究%Study of Heat Stress Pretreatment Effect on Expressions of HSP70,HSF1 in Axial Flap of Rats

    Institute of Scientific and Technical Information of China (English)

    刘哲伟; 苏开新; 陈军; 姚建; 李辉; 王海华; 赵鑫

    2014-01-01

    目的探讨热应激预处理(HSP)对缺血皮瓣组织的保护作用及其与皮瓣中热休克蛋白70(HSP70)和热休克因子1(HSF1)的关系。方法将96只雄性Sprague Dawley 大鼠随机分为缺血组与实验组(HSP+缺血组)。缺血组:皮瓣模型制备后,阻断皮瓣蒂部第1h、2h、4h、6h、8h、10h共6个时间点,每时间点后观测皮瓣存活面积、HSP70、HSF1的变化,并进行统计学分析;实验组( HSP+缺血组):先予以加热预处理,之后与缺血组相同。结果单纯缺血组皮瓣成活率下降率较实验组(HSP+缺血组)明显,差异有显著性(P<0.05),热应激预处理各组皮瓣的HSP70、HSF1合成量与皮瓣存活率呈正相关(r=0.7577,P<0.01)。结论热应激预处理能够改善皮瓣组织的存活率,HSP70、HSF1在热应激预处理改善皮瓣存活率的保护作用中发挥着重要的作用。%Objective This study was to reveal the continuous changes of HSP70 and HSF1 in the delayed and pro-tection of heat stress pretreatment on ischemic preconditioning axial flap in rats. Methods Animals were divided into two groups randomly:HSP+IPC group,ischemic pretreatment group( IPC) . In IPC group,rats skin flaps received ischemic pretreatment for 1 h,2 h,4 h,6 h,8 h,10 h,and the survival rate of the skin flaps,the contents of HSP70 and HSF1 were examined at the 6 time points. HSP+IPC group was given heat stress pretreatment and then the same treatment as IPC group. Results The survival rates of IPC descends more significantly than that of HSR+IPC. The survival rates of skin flaps have positive correlation with the contents of HSP70 and HSF1(r=0. 7577,P<0. 01). Conclusions Heat stress pretreatment can improve the survival rate of ischemia flap and the mechanism of the protection can be related to the func-tion of HSP70 and HSF1.

  3. Non-invasive assessment of animal exercise stress: real-time PCR of GLUT4, COX2, SOD1 and HSP70 in avalanche military dog saliva.

    Science.gov (United States)

    Diverio, S; Guelfi, G; Barbato, O; Di Mari, W; Egidi, M G; Santoro, M M

    2015-01-01

    Exercise has been shown to increase mRNA expression of a growing number of genes. The aim of this study was to assess if mRNA expression of the metabolism- and oxidative stress-related genes GLUT4 (glucose transporter 4), COX2 (cyclooxygenase 2), SOD1 (superoxide dismutase 1) and HSP70 (heat shock protein 70) in saliva changes following acute exercise stress in dogs. For this purpose, 12 avalanche dogs of the Italian Military Force Guardia di Finanza were monitored during simulation of a search for a buried person in an artificial avalanche area. Rectal temperature (RT) and saliva samples were collected the day before the trial (T0), immediately after the descent from a helicopter at the onset of a simulated avalanche search and rescue operation (T1), after the discovery of the buried person (T2) and 2 h later (T3). Expressions of GLUT4, SOD1, COX2 and HSP70 were measured by real-time PCR. The simulated avalanche search and rescue operation was shown to exert a significant effect on RT, as well as on the expression of all metabolism- and oxidative stress-related genes investigated, which peaked at T2. The observed expression patterns indicate an acute exercise stress-induced upregulation, as confirmed by the reductions in expression at T3. Moreover, our findings indicate that saliva is useful for assessing metabolism- and oxidative stress-related genes without the need for restraint, which could affect working dog performance. PMID:25245143

  4. Histopathological alterations and induction of hsp70 in ramshorn snail (Marisa cornuarietis) and zebrafish (Danio rerio) embryos after exposure to PtCl(2).

    Science.gov (United States)

    Osterauer, Raphaela; Köhler, Heinz-R; Triebskorn, Rita

    2010-08-01

    The platinum group metals (PGMs) platinum (Pt), palladium (Pd), and rhodium (Rh) are used in automobile catalytic converters, from which they have been emitted into the environment to an increasing degree during the last 20 years. Despite the bioavailability of these metals to plants and animals, studies determining the effects of PGMs on organisms are extremely rare. In the present study, effects of various concentrations of PtCl(2) (0.1, 1, 10, 50 and 100 microg/L) were investigated with respect to the induction of hsp70 and histopathological alterations in the zebrafish, Danio rerio and the ramshorn snail, Marisa cornuarietis. Histopathological investigations revealed effects of Pt on both species, which varied between slight and strong cellular reactions, depending on the PtCl(2) concentration. The hsp70 level in M. cornuarietis did not show an increase following Pt exposure whereas it was significantly elevated at 100 micorg/L PtCl(2) in D. rerio. PMID:20444508

  5. The effect of grape seed extract and vitamin C feed supplementation on some blood parameters and HSP70 gene expression of broiler chickens suffering from chronic heat stress

    Directory of Open Access Journals (Sweden)

    Hosna Hajati

    2015-07-01

    Full Text Available In this experiment, the effect of hydroalcoholic grape seed extract (GSE and vitamin C feed supplementation on some blood parameters and heat shock protein 70 (HSP70 gene expression of broiler chickens suffering from chronic heat stress was investigated. Experimental diets included control diet (with no additive, 3 levels of GSE (150, 300, 450 mg/kg, and one level of vitamin C (300 mg/kg. Each diet was fed to 5 replicates of 12 male chicks each, from d 1 to 42. The birds suffered from chronic daily heat stress under 34±1°C temperature with 65 to 70% relative humidity for 5 h from 29 to 42 d of age. Results showed that 300 mg/kg GSE supplementation increased body weight of broilers both before and after heat stress condition (at 28 and 42 d, respectively. Also, birds fed 300 mg GSE/kg diet had higher European production efficiency factor during the whole period of the experiment. Supplementation of GSE decreased the concentration of serum glucose at 28 and 42 d; at 42 d (during heat stress condition and at 450 mg/kg diet it decreased cholesterol, triglyceride, lowand very low density lipoprotein concentration of serum blood. Vitamin C supplementation decreased serum cholesterol concentration of broilers suffering from heat stress. HSP70 gene expression in heart and liver of broilers reduced by GSE and vitamin C supplementation pre- and during chronic heat stress condition.

  6. An investigation of molecular dynamics simulation and molecular docking: interaction of citrus flavonoids and bovine β-lactoglobulin in focus.

    Science.gov (United States)

    Sahihi, M; Ghayeb, Y

    2014-08-01

    Citrus flavonoids are natural compounds with important health benefits. The study of their interaction with a transport protein, such as bovine β-lactoglobulin (BLG), at the atomic level could be a valuable factor to control their transport to biological sites. In the present study, molecular docking and molecular dynamics simulation methods were used to investigate the interaction of hesperetin, naringenin, nobiletin and tangeretin as citrus flavonoids and BLG as transport protein. The molecular docking results revealed that these flavonoids bind in the internal cavity of BLG and the BLG affinity for binding the flavonoids follows naringenin>hesperetin>tangeretin>nobiletin. The docking results also indicated that the BLG-flavonoid complexes are stabilized through hydrophobic interactions, hydrogen bond interactions and π-π stacking interactions. The analysis of molecular dynamics (MD) simulation trajectories showed that the root mean square deviation (RMSD) of various systems reaches equilibrium and fluctuates around the mean value at various times. Time evolution of the radius of gyration, total solvent accessible surface of the protein and the second structure of protein showed as well that BLG and BLG-flavonoid complexes were stable around 2500ps, and there was not any conformational change as for BLG-flavonoid complexes. Further, the profiles of atomic fluctuations indicated the rigidity of the ligand binding site during the simulation. PMID:24880994

  7. Cytotoxicity and Expression of c-fos, HSP70, and GADD45/153 Proteins in Human Liver Carcinoma (HepG2 Cells Exposed to Dinitrotoluenes

    Directory of Open Access Journals (Sweden)

    Paul B. Tchounwou

    2005-08-01

    ŽÂ¼g/mL or within the dose range of 0-200 μg/mL for 2,6-DNT. The 45-kDa growth arrest and damage protein was significantly expressed at the dose range of 200 – 250μg/mL for 2,6-DNT and at the dose range of 200 - 400μg/mL for 2,4-DNT. Expression of 153-kDa growth arrest and DNA damage protein was significant at the 100, 200, and 250μg/mL doses for 2,6-DNT and at the 200 μg/mL dose for 2,4-DNT. Overall, these results indicate the potential of DNTs to induce cytotoxic, proteotoxic (HSP70, and genotoxic (GADD45/153 effects, as well as oxidative stress and pro-inflammatory reactions (c-fos.

  8. Review article: Heat shock protein 70 (Hsp70 peptide activated Natural Killer (NK cells for the treatment of patients with non-small cell lung cancer (NSCLC after radiochemotherapy (RCTx – from preclinical studies to a clinical phase II trial

    Directory of Open Access Journals (Sweden)

    Hanno M Specht

    2015-04-01

    Full Text Available Heat shock protein 70 (Hsp70 is frequently overexpressed in tumor cells. An unusual cell surface localization could be demonstrated on a large variety of solid tumors including lung, colorectal, breast, squamous cell carcinomas of the head and neck, prostate and pancreatic carcinomas, glioblastomas, sarcomas and hematological malignancies, but not on corresponding normal tissues. A membrane (mHsp70-positive phenotype can be determined either directly on single cell suspensions of tumor biopsies by flow cytometry using cmHsp70.1 monoclonal antibody or indirectly in the serum of patients using a novel lipHsp70 ELISA. A mHsp70-positive tumor phenotype has been associated with highly aggressive tumors, causing invasion and metastases and resistance to cell death. However, natural killer (NK, but not T cells were found to kill mHsp70-positive tumor cells after activation with a naturally occurring Hsp70 peptide (TKD plus low dose IL-2 (TKD/IL-2. Safety and tolerability of ex vivo TKD/IL-2 stimulated, autologous NK cells has been demonstrated in patients with metastasized colorectal and NSCLC in a phase I clinical trial. Based on promising clinical results of the previous study, a phase II randomized clinical study was initiated in 2015. The primary objective of this multicenter proof-of-concept trial is to examine whether an adjuvant treatment of NSCLC patients after platinum based radiochemotherapy with TKD/IL-2 activated, autologous NK cells is clinically effective. As a mHsp70-positive tumor phenotype is associated with poor clinical outcome only mHsp70-positive tumor patients will be recruited into the trial. The primary endpoint of this study will be the comparison of the progression-free survival of patients treated with ex vivo activated NK cells compared to patients who were treated with radiochemotherapy alone. As secondary endpoints overall survival, toxicity, quality-of-life and biological responses will be determined in both study groups.

  9. In contrast to conventional inactivated influenza vaccines, 4xM2e.HSP70c fusion protein fully protected mice against lethal dose of H1, H3 and H9 influenza A isolates circulating in Iran

    Energy Technology Data Exchange (ETDEWEB)

    Ebrahimi, Seyyed Mahmoud, E-mail: smebrahimi@shirazu.ac.ir [Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, P.O. Box 14155-3651,Tehran (Iran, Islamic Republic of); Research Center of Virus and Vaccine, Baqiyatallah University of Medical Science, P.O.Box 14155-3651, Tehran (Iran, Islamic Republic of); Dabaghian, Mehran [Department of Pathobiology, University of Tehran, Faculty of Veterinary Medicine, P.O. Box 14155-6453, Tehran (Iran, Islamic Republic of); Tebianian, Majid [Department of Biotechnology, Razi Vaccine and Serum Research Institute (RVSRI), P.O. Box 31975/148, Karaj, Tehran (Iran, Islamic Republic of); Zabeh Jazi, Mohammad Hossein [Department of Pathobiology, University of Tehran, Faculty of Veterinary Medicine, P.O. Box 14155-6453, Tehran (Iran, Islamic Republic of)

    2012-08-15

    Ideal vaccines against influenza viruses should elicit not only a humoral response, but also a cellular response. Mycobacterium tuberculosis HSP70 (mHSP70) have been found to promote immunogenic APCs function, elicit a strong cytotoxic T lymphocyte (CTL) response, and prevent the induction of tolerance. Moreover, it showed linkage of antigens to the C-terminus of mHSP70 (mHSP70c) can represent them as vaccines resulted in more potent, protective antigen specific responses in the absence of adjuvants or complex formulations. Hence, recombinant fusion protein comprising C-terminus of mHSP70 genetically fused to four tandem repeats of the ectodomain of the conserved influenza matrix protein M2 (M2e) was expressed in Escherichia coli, purified under denaturing condition, refolding, and then confirmed by SDS-PAGE, respectively. The recombinant fusion protein, 4xM2e.HSP70c, retained its immunogenicity and displayed the protective epitope of M2e by ELISA and FITC assays. A prime-boost administration of 4xM2e.HSP70c formulated in F105 buffer by intramuscular route in mice (Balb/C) provided full protection against lethal dose of mouse-adapted H1N1, H3N2, or H9N2 influenza A isolates from Iran compared to 0-33.34% survival rate of challenged unimmunized and immunized mice with the currently in use conventional vaccines designated as control groups. However, protection induced by immunization with 4xM2e.HSP70c failed to prevent weight loss in challenged mice; they experienced significantly lower weight loss, clinical symptoms and higher lung viral clearance in comparison with protective effects of conventional influenza vaccines in challenged mice. These data demonstrate that C-terminal domain of mHSP70 can be a superior candidate to deliver the adjuvant function in M2e-based influenza A vaccine in order to provide significant protection against multiple influenza A virus strains.

  10. In contrast to conventional inactivated influenza vaccines, 4xM2e.HSP70c fusion protein fully protected mice against lethal dose of H1, H3 and H9 influenza A isolates circulating in Iran

    International Nuclear Information System (INIS)

    Ideal vaccines against influenza viruses should elicit not only a humoral response, but also a cellular response. Mycobacterium tuberculosis HSP70 (mHSP70) have been found to promote immunogenic APCs function, elicit a strong cytotoxic T lymphocyte (CTL) response, and prevent the induction of tolerance. Moreover, it showed linkage of antigens to the C-terminus of mHSP70 (mHSP70c) can represent them as vaccines resulted in more potent, protective antigen specific responses in the absence of adjuvants or complex formulations. Hence, recombinant fusion protein comprising C-terminus of mHSP70 genetically fused to four tandem repeats of the ectodomain of the conserved influenza matrix protein M2 (M2e) was expressed in Escherichia coli, purified under denaturing condition, refolding, and then confirmed by SDS–PAGE, respectively. The recombinant fusion protein, 4xM2e.HSP70c, retained its immunogenicity and displayed the protective epitope of M2e by ELISA and FITC assays. A prime-boost administration of 4xM2e.HSP70c formulated in F105 buffer by intramuscular route in mice (Balb/C) provided full protection against lethal dose of mouse-adapted H1N1, H3N2, or H9N2 influenza A isolates from Iran compared to 0–33.34% survival rate of challenged unimmunized and immunized mice with the currently in use conventional vaccines designated as control groups. However, protection induced by immunization with 4xM2e.HSP70c failed to prevent weight loss in challenged mice; they experienced significantly lower weight loss, clinical symptoms and higher lung viral clearance in comparison with protective effects of conventional influenza vaccines in challenged mice. These data demonstrate that C-terminal domain of mHSP70 can be a superior candidate to deliver the adjuvant function in M2e-based influenza A vaccine in order to provide significant protection against multiple influenza A virus strains.

  11. 乐至黑山羊与藏山羊HSC70、HSP70.1和HSF1基因的mRNA表达差异分析%Analysis of mRNA Differential Expression of HSC70、HSP70.1 and HSF1 Gene in Lezhi Black Goat and Tibetan Goat

    Institute of Scientific and Technical Information of China (English)

    牛惠然; 字向东; 任陶; 王永; 任称罗尔日; 朱万岭; 付锡三

    2013-01-01

    In order to investigate the relationship between eco-adaptation and heat shock protein70, the heat shock cognate protein 70 (HSC70), heat shock protein 70. 1 (HSP70. 1) and the heat shock transcription factor 1 (HSF1) genes in five tissue samples of Lezhi black goat and Tibetan goat were determined by qPCR. The results showed that the expression of HSC70 mRNA in Tibetan goat's heart was 4. 5 times to that of the Lezhi black goat (P<0. 05) , but in pituitary, Lezhi black goat is 3. 7 times to Tibetan goat. Its expression in the ovary, liver, uterus, was not different between the two breeds. The expression of HSF1 mRNA in Tibetan goat ovarian, heart was 7. 9, 3. 1 times to that of the Lezhi black goat (F< 0. 05), however, in the liver and uterus tissue samples, Lezhi black goat was in high expression levels than Tibetan goat (6. 3 fold, 10. 4 fold; P<0. 05). The expression of HSP70. 1 mRNA in Lezhi black goat pituitary was 6. 8 times to Tibetan goat (P<0. 05). It could be concluded that the difference in the expression of HSC70, HSP70. 1 and HSF1 wight be related to the eco-environment and rearing methods (grazing) in Lezhi black goat and Tibetan goat.%试验旨在利用qPCR技术对热休克同源蛋白(HSC70)、热休克蛋白70.1(HSP70.1)和热休克转录因子1(HSF1)基因在乐至黑山羊和藏山羊五个组织样中的表达量进行研究,以探讨热休克蛋白家族70 (HSP70)与山羊生态适应性的关系.结果表明,HSC70mRNA在藏山羊心脏中的表达量是乐至黑山羊中的4.5倍(P<0.05),但在脑垂体中乐至黑山羊的表达量是藏山羊的3.7倍(P<0.05),在卵巢、肝脏和子宫中两品种间无显著差异;HSF1 mRNA在藏山羊卵巢、心脏中的表达量分别是乐至黑山羊的7.9倍和3.1倍(P<0.05),在肝脏和子宫中乐至黑山羊的表达量却分别是藏山羊的6.3倍和10.4倍(P<0.05);HSP70.1 mRNA在乐至黑山羊脑垂体中的表达量是藏山羊的6.8倍(P<0.05).说明HSC70、HSP70.1

  12. Combined effects of thermal stress and Cd on lysosomal biomarkers and transcription of genes encoding lysosomal enzymes and HSP70 in mussels, Mytilus galloprovincialis

    Energy Technology Data Exchange (ETDEWEB)

    Izagirre, Urtzi; Errasti, Aitzpea; Bilbao, Eider; Múgica, María; Marigómez, Ionan, E-mail: ionan.marigomez@ehu.es

    2014-04-01

    Highlights: • Thermal stress and Cd caused lysosomal enlargement and membrane destabilisation. • hex, gusb and ctsl but not hsp70 were up-regulated at elevated temperature but down-regulated by Cd. • Thermal stress influenced lysosomal responses to Cd exposure. • The presence of Cd jeopardised responsiveness against thermal stress. - Abstract: In estuaries and coastal areas, intertidal organisms may be subject to thermal stress resulting from global warming, together with pollution. In the present study, the combined effects of thermal stress and exposure to Cd were investigated in the endo-lysosomal system of digestive cells in mussels, Mytilus galloprovincialis. Mussels were maintained for 24 h at 18 °C and 26 °C seawater temperature in absence and presence of 50 μg Cd/L seawater. Cadmium accumulation in digestive gland tissue, lysosomal structural changes and membrane stability were determined. Semi-quantitative PCR was applied to reveal the changes elicited by the different experimental conditions in hexosaminidase (hex), β-glucuronidase (gusb), cathepsin L (ctsl) and heat shock protein 70 (hsp70) gene transcription levels. Thermal stress provoked lysosomal enlargement whilst Cd-exposure led to fusion of lysosomes. Both thermal stress and Cd-exposure caused lysosomal membrane destabilisation. hex, gusb and ctsl genes but not hsp70 gene were transcriptionally up-regulated as a result of thermal stress. In contrast, all the studied genes were transcriptionally down-regulated in response to Cd-exposure. Cd bioaccumulation was comparable at 18 °C and 26 °C seawater temperatures but interactions between thermal stress and Cd-exposure were remarkable both in lysosomal biomarkers and in gene transcription. hex, gusb and ctsl genes, reacted to elevated temperature in absence of Cd but not in Cd-exposed mussels. Therefore, thermal stress resulting from global warming might influence the use and interpretation of lysosomal biomarkers in marine pollution

  13. Fractionated radiotherapy is the main stimulus for the induction of cell death and of Hsp70 release of p53 mutated glioblastoma cell lines

    International Nuclear Information System (INIS)

    Glioblastoma multiforme (GBM) is the most common primary brain tumor in adults. Despite a multimodal therapy consisting of resection followed by fractionated radiotherapy (RT) combined with the chemotherapeutic agent (CT) temozolomide (TMZ), its recurrence is almost inevitable. Since the immune system is capable of eliminating small tumor masses, a therapy should also aim to stimulate anti-tumor immune responses by induction of immunogenic cell death forms. The histone deacetylase inhibitor valproic acid (VPA) might foster this. Reflecting therapy standards, we applied in our in vitro model fractionated RT with a single dose of 2Gy and clinically relevant concentrations of CT. Not only the impact of RT and/or CT with TMZ and/or VPA on the clonogenic potential and cell cycle of the glioblastoma cell lines T98G, U251MG, and U87MG was analyzed, but also the resulting cell death forms and release of danger signals such as heat-shock protein70 (Hsp70) and high-mobility group protein B1 (HMGB1). The clonogenic assays revealed that T98G and U251MG, having mutated tumor suppressor protein p53, are more resistant to RT and CT than U87MG with wild type (WT) p53. In all glioblastoma cells lines, fractionated RT induced a G2 cell cycle arrest, but only in the case of U87MG, TMZ and/or VPA alone resulted in this cell cycle block. Further, fractionated RT significantly increased the number of apoptotic and necrotic tumor cells in all three cell lines. However, only in U87MG, the treatment with TMZ and/or VPA alone, or in combination with fractionated RT, induced significantly more cell death compared to untreated or irradiated controls. While necrotic glioblastoma cells were present after VPA, TMZ especially led to significantly increased amounts of U87MG cells in the radiosensitive G2 cell cycle phase. While CT did not impact on the release of Hsp70, fractionated RT resulted in significantly increased extracellular concentrations of Hsp70 in p53 mutated and WT glioblastoma

  14. Mice chronically exposed to low dose ionizing radiation possess splenocytes with elevated levels of HSP70 mRNA, HSC70 and HSP72 and with an increased capacity to proliferate

    International Nuclear Information System (INIS)

    The purpose of this study was to determine whether the enhanced proliferative activity of splenocytes induced by exposing mice to whole body, chronic, intermittent low doses of ionizing radiation is associated with an increase in the expression of stress protein genes. Mice were exposed to a γ-irradiation protocol of 0, 0.04 or 0.10 Gy/day for 5 consequent days/week, for 4 weeks. Splenocytes were then assessed for their levels of heat shock protein (HSP) 70 mRNA, glyceraldehyde 3-phosphate dehydrogenase (GAPD) mRNA, HSC70 (a constitutively-expressed isoform of HSP70) and HSP72 (an inducible isoform of HSP70), before and 1 day after mitogenic stimulation. (author)

  15. 亚硝酸盐急性胁迫对中华绒螯蟹幼体相关免疫指标和应激蛋白(HSP70)表达的影响%Effects of Acute Nitrite Exposure on Immunity Indicators and HSP70 Expression in Chinese Mitten-hand Crab (Eriocheir sinensis)

    Institute of Scientific and Technical Information of China (English)

    洪美玲; 陈立侨; 孙新谨; 李二超; 顾顺樟; 禹娜

    2011-01-01

    为探讨亚硝酸盐急性胁迫对中华绒螯蟹机体免疫力的影响,以及机体在胁迫过程中的应对策略,分别测定了幼蟹(平均体重3.25 g)在10、20、30、40 mg L-1业硝酸盐暴露组和对照组中血细胞总数(Total hemocyte counts,THC)、超氧化物歧化酶( Superoxide dismutase,SOD)活力、丙二醛(Malodiadchydc,MDA)含量以及热休克蛋白 70( Heat stress protein 70,HSP70)表达量在受胁迫2d内的动态变化.结果表明:1)低浓度亚硝酸盐(10 mg L-1)胁迫组THC在胁迫3h时显著高于对照组,肝胰腺中SOD活性在胁迫3h和6h时亦显著提高,表现出一定的“毒物兴奋效应”2)但随着暴露时间的延长和亚硝酸盐浓度的升高,幼蟹THC减少,肝胰腺SOD活性下降,而MDA含量升高3)对照组中有少量的HSP70表达.幼蟹经10、20 mg L-1亚硝酸盐处理后,鳃组织中HSP70表达量在胁迫6h后达到最大,而经30、40 mg L-1 亚硝酸盐处理后,HSP70表达量在胁迫3h后达到最大.各浓度组所诱导的HSP70最大表达量与水中亚硝酸盐的浓度呈正相关.以上结果说明亚硝酸盐胁迫可通过加剧组织中脂质过氧化的方式导致机体免疫力下降,肝胰腺中MDA的含量变化可作为业硝酸盐胁迫的有效评价指标之一,而HSP70表达量的变化可作为早期预警指标应用于亚硝酸盐胁迫评价中.图2表1参36%Nitrite is an intermediate product either in the bacterial nitrification of ammonia, or in the bacterial denitrification of nitrate. The concentration of nitrite may reach 50 mg L-1 in intensive aquacultural systems due to an imbalance in biofilter resulting from overcrowding effect and poor water recirculation. High levels of nitrite in water were potential factors triggering stress on aquatic organisms. The sensitive indicators of stress evaluation for the Chinese mitten-hand crab Eriocheir smensis were selected to investigate the effect of ambient nitrite on the immunity response. In this experiment, the

  16. BAG-1/SODD, HSP70, and HSP90 are potential prognostic markers of poor survival in node-negative breast carcinoma.

    Science.gov (United States)

    Davidson, Ben; Valborg Reinertsen, Kristin; Trinh, Don; Reed, Wenche; Bøhler, Per Johannes

    2016-08-01

    The objective of this study was to analyze the expression and clinical role of 13 signaling molecules in a large cohort of breast carcinoma patients with long follow-up period. Breast carcinomas (n=410) were analyzed for protein expression of phosphorylated mitogen-activated protein kinases (p-ERK, p-JNK, p-p38) and phosphoinositide 3-kinase signaling pathway proteins (p-AKT, p-mTOR, p-p70S6K); the BAG family proteins BAG-1 and BAG-4/SODD; the antiapoptotic protein Bcl-2; the inhibitor of apoptosis family member Survivin; and the heat shock protein family members HSP27, HSP70, and HSP90. Protein expression was studied for association with clinicopathological parameters and survival. Significantly higher expression of p-AKT (Pmarkers in node-negative breast carcinoma that merit further research. PMID:27038683

  17. Toxic Potential of Synthesized Graphene Zinc Oxide Nanocomposite in the Third Instar Larvae of Transgenic Drosophila melanogaster (hsp70-lacZBg9

    Directory of Open Access Journals (Sweden)

    Yasir Hasan Siddique

    2014-01-01

    Full Text Available In the present study the graphene zinc oxide nanocomposite (GZNC was synthesized, characterized, and evaluated for its toxic potential on third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZBg9. The synthesized GZNC was characterized by X-ray diffraction (XRD, Fourier transform infrared spectroscopy (FTIR, thermogravimetric analysis (TGA, scanning electron microscopy (SEM, and transmission electron microscopy (TEM. The GZNC in 0.1% dimethyl sulphoxide (DMSO was sonicated for 10 minutes and the final concentrations 0.033, 0.099, 0.199, and 3.996 μg/μL of diet were established. The third instar larvae were allowed to feed on it separately for 24 and 48 hr. The hsp70 expression was measured by o-nitrophenyl-β-D-galactopyranoside assay, tissue damage was measured by trypan blue exclusion test, and β-galactosidase activity was monitored by in situ histochemical β-galactosidase staining. Oxidative stress was monitored by performing lipid peroxidation assay and total protein estimation. Ethidium bromide/acridine orange staining was performed on midgut cells for apoptotic index and the comet assay was performed for the DNA damage. The results of the present study showed that the exposure of 0.199 and 3.996 μg/μL of GZNC was toxic for both 24 hr and 48 hr of exposure. The doses of 0.033 μg/μL and 0.099 of GZNC showed no toxic effects on its exposure to the third instar larvae for 24 hr as well as 48 hr of duration.

  18. Expression of HSP70 family and HSP90 family in skeletal muscles of transport stressed pigs%运输应激猪骨骼肌中热应激蛋白HSP70和HSP90家族的表达

    Institute of Scientific and Technical Information of China (English)

    鲍恩东; Sultan,KR; 等

    2001-01-01

    To demonstrate the expression of heat shock protein s(HSP)in skeletal muscles and to estimate the relationship between expression of HSP and pork quality, using Western blot four kinds of HSP which belongs to HSP70 family and HSP90 family respectively in porcine skeletal muscle tissues were determined after a long distance transport.All four HSP,namely HSP70,HSP72,HSP86 and HSP90,were regularly detected in M.longissimus dorsi and M.gluteus medius of both the transported and the cont rol group.The expression of HSP in control pigs showed that HSPs have some indep endent physiological from stress response undetermined functions.after 6 h trans port,the induction of the HSP70 and HSP72 in skeletal muscle tissues was slightly elevated(P>0.05).However,expression of the HSP 86 and the HSP90,especially HSP90,markedly decreased(P<0.01)in skelet al muscles.The results are implied that the reduction tendency of HSP90 fa mily is relative to pork quality and may serve as a potential marker for stress response.%利用Western blot技术,检测长途运输试验猪骨骼肌(背最长肌和臀大肌)中分属于HSP70家族和HSP90家族的4种应激蛋白(HSP70,HSP 72,HSP86和HSP90)的表达。所有运输应激猪和对照猪肌肉组织中均检测到了上述4种HSP。对照猪组织中HSP的表达说明HSP除了在受应激的细胞内行使生理功能外,还具有独立于应激刺激应答以外的其它作用。6 h的长途运输后,HSP的表达量明显不同,HSP70和HSP72在肌肉组织中的表达虽然有一定的增加,但统计学分析差异不显著(P>0.05);而HSP86和HSP90在骨骼肌中的表达明显下降,尤以HS P90下降最为显著(P<0.01)。提示HSP90家族成员与肉品品质相关,可能作为判断应激损伤的指征。

  19. 大鼠再生肝中hsbp1、hsf1、hsf2、hsp70表达水平改变的分析%ANALYSIS OF CHANGES ABOUT hsbp1, hsf1, hsf2 AND hsp70'S EXPRESSION LEVELS IN RAT'S REGENERATING LIVER

    Institute of Scientific and Technical Information of China (English)

    苏丽娟; 常翠芳; 韩鸿鹏; 马辉; 徐存拴

    2006-01-01

    在克隆了大鼠热休克因子结合蛋白1基因(hsbp1)全长cDNA基础上,进一步分析它在肝再生中作用.用SD纯系大鼠为材料,按Higgens等方法建立大鼠部分肝切除(PH)模型;用原位杂交等方法分析hsbp1在肝再生中表达变化;用基因表达谱芯片分析hsbp1、hsf1、hsf2和hsp70在肝再生中表达变化.原位杂交和基因表达谱芯片分析表明,PH后6h和66-144h,hsp1表达发生了有意义上调;8-16h,hsf1表达发生了有意义上调;2-16h,hsf2表达发生了有意义上调;0.5-24h,hsp70表达发生了有意义上调.假手术(只打开腹腔和翻动肝叶,但不进行部分肝切除)后0.5-2h,hsbp1表达发生了有意义下调;8-16h,hsf1表达发生了有意义上调;0-144h,hsf2未发生有意义表达变化;0.5-30h,hsp70表达发生了有意义上调.根据实验结果推测,PH后hsbp1表达上调可增加细胞内HSBP1量,促进生长、发育、分化相关基因表达和再生肝的组织结构功能重建;(假)手术后hsbp1表达下调可减少细胞内HSBP1量,有利于HSF1上调hsp70表达,提高机体和肝脏抗损伤能力.

  20. Identification and Molecular Characterization of Nuclear Citrus leprosis virus, a Member of the Proposed Dichorhavirus Genus Infecting Multiple Citrus Species in Mexico.

    Science.gov (United States)

    Roy, Avijit; Stone, Andrew L; Shao, Jonathan; Otero-Colina, Gabriel; Wei, Gang; Choudhary, Nandlal; Achor, Diann; Levy, Laurene; Nakhla, Mark K; Hartung, John S; Schneider, William L; Brlansky, Ronald H

    2015-04-01

    Citrus leprosis is one of the most destructive diseases of Citrus spp. and is associated with two unrelated virus groups that produce particles primarily in either the cytoplasm or nucleus of infected plant cells. Symptoms of leprosis, including chlorotic spots surrounded by yellow haloes on leaves and necrotic spots on twigs and fruit, were observed on leprosis-affected mandarin and navel sweet orange trees in the state of Querétaro, Mexico. Serological and molecular assays showed that the cytoplasmic types of Citrus leprosis virus (CiLV-C) often associated with leprosis symptomatic tissues were absent. However, using transmission electron microscopy, bullet-shaped rhabdovirus-like virions were observed in the nuclei and cytoplasm of the citrus leprosis-infected leaf tissues. An analysis of small RNA populations from symptomatic tissue was carried out to determine the genome sequence of the rhabdovirus-like particles observed in the citrus leprosis samples. The complete genome sequence showed that the nuclear type of CiLV (CiLV-N) present in the samples consisted of two negative-sense RNAs: 6,268-nucleotide (nt)-long RNA1 and 5,847-nt-long RNA2, excluding the poly(A) tails. CiLV-N had a genome organization identical to that of Orchid fleck virus (OFV), with the exception of shorter 5' untranslated regions in RNA1 (53 versus 205 nt) and RNA2 (34 versus 182 nt). Phylogenetic trees constructed with the amino acid sequences of the nucleocapsid (N) and glycoproteins (G) and the RNA polymerase (L protein) showed that CiLV-N clusters with OFV. Furthermore, phylogenetic analyses of N protein established CiLV-N as a member of the proposed genus Dichorhavirus. Reverse-transcription polymerase chain reaction primers for the detection of CiLV-N were designed based on the sequence of the N gene and the assay was optimized and tested to detect the presence of CiLV-N in both diseased and symptom-free plants. PMID:25423071

  1. Mekanisme Peningkatan Heat Shock Protein-70 pada Kanker Payudara Tikus yang Diradiasi, Pascapemberian Ekstrak Meniran (Phyllanthus niruri (MECHANISM OF INCREASING OF HSP-70 ON IRRADIATED RAT BREAST CANCER, DUE TO APPLICATION OF EXTRACT OF PHYLLANTHUS NIR

    Directory of Open Access Journals (Sweden)

    Bambang Soeprijanto

    2014-10-01

    Full Text Available The used of radiation as a cancer treatment is also proven giving the side effect of damaging thenormal tissue. The extract of Phyllanthus niruri L plant has already been known have an ability to modulatethe immune system. Heat Shock Protein 70 (HSP70 is a protein which can protect other proteins from anydamages. The transcription factor, such as Nuclear factor kappa-light-chain-enhancer of activated B cells(NfkB,is required for protein synthesis. This research was intended to analyze the mechanism, of the immunecompetent cells in expressing the HSP70,through the increase of Nf-kB, at the rat breast cancer tissueunder radiation, due to the application of the extract of P.niruri L plant. An experimental study wasperformed by using the randomized separate pre-test post test controlled group design. The female whiterat (Rattus norvegicus strain Sprague Dawleystrain, undergoing breast cancer due to the application ofcarcinogen materials 7,12-dimethylbenz(aantrasen(DMBA at 20 mg/kg.b.wt, and then the externalradiation at 6Gy given. The treatment group was given the aqueous extract of P.niruri L plant per oral at250 mg/kg b.wt.By immunohistochemistry and t-test study we observe a significant increases in thenumber of cells expressing Nf-kB (p<0.05 and a significant increases in the number of cells expressingHSP70 (p<0.05 at the treatment group. At the regression test, there found to be a stronger influence of NfkBto HSP70 at the treatment group. It is concluded that the mechanism of cell increase expressing theHSP70 at the rat breast cancer tissue under radiation due to the application of the aqueous extract ofP.niruri L plant per oral, is through the increase of cells expressing the Nf-kB.

  2. The human HSP70/HSP40 chaperone family: a study on its capacity to combat proteotoxic stress

    OpenAIRE

    Hageman, Jurre

    2008-01-01

    All cells are constantly threatened by both physical and chemical stress factors that can damage e.g. the DNA and proteins. Whereas the molecular mechanisms responsible for DNA repair have been studied in great detail, the mechanism underlying the repair and disposal of damaged proteins are far less investigated and less understood. It is well known that DNA damage in combination with a malfunctioning repair mechanism may ultimately lead to diseases such as cancer. But also damaged proteins a...

  3. Mitochondrial Heat Shock Protein Machinery Hsp70/Hsp40 Is Indispensable for Proper Mitochondrial DNA Maintenance and Replication

    Czech Academy of Sciences Publication Activity Database

    Týč, Jiří; Klingbeil, M.M.; Lukeš, Julius

    2015-01-01

    Roč. 6, č. 1 (2015), e02425-14. ISSN 2150-7511 R&D Projects: GA MŠk LH12104; GA MŠk(CZ) EE2.3.30.0032; GA ČR GAP305/12/2261 EU Projects: European Commission(XE) 316304 Institutional support: RVO:60077344 Keywords : procyclic Trypanosoma brucei * transfer RNA import * kinetoplast DNA Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.786, year: 2014

  4. Gastroprotective activity of Annona muricata leaves against ethanol-induced gastric injury in rats via Hsp70/Bax involvement

    Directory of Open Access Journals (Sweden)

    Moghadamtousi SZ

    2014-10-01

    downregulation of Bax and upregulation of Hsp70 proteins after pretreatment. Collectively, the present results suggest that EEAM has a promising antiulcer potential, which could be attributed to its suppressive effect against oxidative damage and preservative effect toward gastric wall mucus. Keywords: Annona muricata, annonaceae, gastric injury, antioxidants, Hsp70/Bax

  5. Involvement of NF-ΚB and HSP70 signaling pathways in the apoptosis of MDA-MB-231 cells induced by a prenylated xanthone compound, α-mangostin, from Cratoxylum arborescens

    Directory of Open Access Journals (Sweden)

    Ibrahim MY

    2014-11-01

    the role of the central apoptosis-related proteins, a protein array followed by immunoblot analysis was conducted. Moreover, the involvement of nuclear factor-kappa B (NF-ΚB was also analyzed. Results: Apoptosis was confirmed by the apoptotic cells stained with annexin V and increase in chromatin condensation in nucleus. Treatment of cells with AM promoted cell death-transducing signals that reduced MMP by downregulation of Bcl-2 and upregulation of Bax, triggering cytochrome c release from the mitochondria to the cytosol. The released cytochrome c triggered the activation of caspase-9 followed by the executioner caspase-3/7 and then cleaved the PARP protein. Increase of caspase-8 showed the involvement of extrinsic pathway. AM treatment significantly arrested the cells at the S phase (P<0.05 concomitant with an increase in reactive oxygen species. The protein array and Western blotting demonstrated the expression of HSP70. Moreover, AM significantly blocked the induced translocation of NF-ΚB from cytoplasm to nucleus. Conclusion: Together, the results demonstrate that the AM isolated from C. arborescens inhibited the proliferation of MDA-MB-231 cells, leading to cell cycle arrest and programmed cell death, which was suggested to occur through both the extrinsic and intrinsic apoptosis pathways with involvement of the NF-ΚB and HSP70 signaling pathways. Keywords: mitochondria, protein array, caspase-3/7

  6. Molecular Detection of Spiroplasma Citri Associated with Stubborn Disease in Citrus Orchards in Syria

    Science.gov (United States)

    Spiroplasma citri, a phloem-limited pathogen, causes citrus stubborn disease (CSD) and can be transmitted from plant to plant by several species of phloem-feeding leafhoppers. CSD is an important disorder in certain warm and arid citrus-growing areas, and its agent has been recorded from several Med...

  7. The effects of heat shock induction expression of HSP70 on hprt genetic mutation induced by X-ray in lymphocytes

    International Nuclear Information System (INIS)

    Objective: To explore the effects of heat shock pretreatment on hprt genetic mutation induced by X-ray in lymphocytes. Method: The rat peripheral lymphocytes were cultured under 42 degree C and 5% CO2 for 90 min, then the cells were divided into 4 groups (control group, heat shock, radiation group, heat shock + radiation group), each group including 6 parallel samples. And each group was fractionated into 2 parts, one part with 6-TG (final concentration was 0.1 mmol/L) and the other without 6-TG. Each group of cells were irradiated by 0.5 Gy, 1.0 Gy, 1.5 Gy, 2.0 Gy, 2.5 Gy, 3.0 Gy X-ray and recovery cultured for 3 h, 6 h, 10 h, and then polykaryocytes test was used to detect the hprt mutation in the rat peripheral lymphocytes. Results: After exposure to different dose X-ray and recovery culture for 3 h, 6 h, 10 h, the mutation rate of hprt in heat shock + radiation group was significantly decreased (P<0.01) compared to radiation group. Conclusion: Heat shock pretreatment has protection effects on hprt mutation induced by X-ray. HSP70 has more obvious protection effects in low dose radiation than high dose radiation on hprt mutation induced by X-ray (P<0.01). There is negative correlation between protection effects and radiation doses. (authors)

  8. Immunohistochemical evaluation of expression of heat shock proteins HSP70 and HSP90 in mammary gland neoplasms in bitches.

    Science.gov (United States)

    Badowska-Kozakiewicz, A M; Malicka, E

    2012-01-01

    Heat shock proteins have essential roles in a number of pathophysiologic conditions including carcinogenesis and represent a group of novel molecular markers in cancer management. The aim of this study was to investigate heat shock protein expression in correlation with other neoplasm traits such as: histological type, differentiation grade, proliferative activity, estrogenic receptor expression, and cyclooxygenase-2 and p53 proteins. Material for the investigation comprised 133 tumors of the mammary gland collected from bitches. In total 14 adenomas, 66 complex carcinomas, 47 simple carcinomas and 6 solid carcinomas were collected. Evaluations were conducted with histopathological and immunohistochemical methods using suitable antibodies. Expression of heat shock protein 70 was observed in all types of evaluated neoplasms. A higher average number of cells undergoing expression of heat shock protein 70, which was statistically insignificant, was established in complex and simple cancers and in cancers with the 1st and the 2nd degree of histological malignancy. Expression of heat shock protein 90 was observed in all studied neoplasms; it was very insignificant in adenomas, compared to cancers, and the highest expression was established in the solid cancers, as well as in cancers with the 2nd degree of histological malignancy. This high expression of heat shock protein 90 was correlated with proliferative activity. The results suggest that heat shock protein 90 is involved in canine mammary gland carcinogenesis. The results also suggest that heat shock protein 90 may be a prognostic factor, but this requires detailed clinical confirmation. PMID:22844695

  9. The Mechanism of Calcium and Calcium-activated Calmodulin Modulating the Expression of Hsp70%钙-钙调素信号通路调节热应激蛋白70表达的机制

    Institute of Scientific and Technical Information of China (English)

    齐晓楠; 赵园; 田文儒

    2015-01-01

    随着温室效应的加剧,全球气候变暖已经成为现代畜牧业生产体系所面临的严峻挑战.高温炎热气候已成为影响畜禽生长繁殖的一种主要的非生物胁迫.热应激刺激时,动物体产生一系列生理变化来提高机体的耐受性,而本文旨在阐述钙(calcium,Ca2+)-钙调蛋白(calcium-activated calmodulin,CaM)参与的耐热信号的转导机制,即热应激蛋白(heat shock proteins,Hsps)的表达机制.热应激时,细胞内Ca2+浓度上升,激活CaM及其表达量的增加,随即CaM作用于其下游的2种靶蛋白,即2条信号转导支路来提高热激转录因子l(heat shock factor 1,HSFl)的转录活性,进而调节热应激蛋白70(Hsp70)的表达.热应激时,Hsps表达的2条具体通路:①Ca2+-CaM激活下游的靶蛋白—钙/钙调素依赖性蛋白激酶Ⅱ(Ca2+/calmodulin-dependent protein kinaseⅡ,CaMKⅡ),CaMKⅡ通过促进HSF1(230)的磷酸化来提高HSF1的转录活性,进而诱导Hsp70的表达;②Ca2+-CaM激活下游另一个靶蛋白Hsp70,Hsp70通过与增多的CaM形成复合物,释放HSF1单体,增多的HSF1形成有活性的HSF1三聚体,并入核与热激元件(heat shockelement,HSE)结合,使HSF1具有转录活性,进而诱导Hsp70的表达.本文系统完整地综述了Ca2+-CaM-HSF1-Hsp70热应激信号系统,在感受、传导和响应热应激刺激时的机制.

  10. Protective effect of SjC23-Hsp70 DNA vaccine and interleukin-12 on Schistosoma japonicum infection in water buffalos%日本血吸虫SjC23-Hsp70 DNA疫苗与IL-12对水牛保护性作用的研究

    Institute of Scientific and Technical Information of China (English)

    胡平成; 夏大; 崔虹艳; 张苹芳; 何永康; 喻鑫玲; 孙振球

    2012-01-01

    目的:研究日本血吸虫中国大陆株23 kD膜蛋白-热休克蛋白(SjC23-Hsp70)DNA疫苗联合佐剂白细胞介素12(IL-12)质粒DNA对水牛的免疫保护作用.方法:将血吸虫病非流行区8~10月龄健康水牛45头随机分为A组(SjC23-Hsp70+IL-12)、B组(SjC23+IL- 12)和C组(pVAX+IL-12),每组15头.每头牛经肩部肌注免疫3次,每次间隔28 d.末次免疫后28 d,每头牛感染日本血吸虫尾蚴1000条.解剖前2天及当天分别收集粪便1次,用定量法检测虫卵和毛蚴数.攻击感染后56天解剖所有水牛,经胸主动脉灌冲法收集成虫,计数成虫数,检测每克肝组织虫卵数.结果:A,B组与C组相比,分别获得45.70%和26.61%的减雌率,44.51%和25.84%的减虫率,41.10%和31.63%的减粪卵率,48.11%和38.07%的减毛蚴率及43.39%和31.95%的减肝卵率.A组的5个率均比B组高(P<0.05).结论:用SjC23-Hsp70 DNA疫苗和IL-12联合免疫水牛可获得明显的免疫保护作用.%Objective: To determine the immune-protective effect of Japan Schistosoma (Chinese mainland strain) 23 kD membrane protein-heat shock protein (SjC23-Hsp70) DNA vaccine plus adjuvant-induced interleukin-12 (IL-12) plasmid DNA on Schistosoma japonicum infection in water buffalos. Methods: Forty-five health water buffalos (8-10 months old) in non-endemic area of schistosomiasis were randomly assigned into group A (SjC23-Hsp70+IL-12, 300 μg), group B (SjC23+IL-12, 300 μg) and group C (pVAX+IL-12, 300 μg), 15 in each group. Each buffalo was immuned by shoulder intramuscular injection for 3 times, at an interval of 28 days. Twenty-eight days after the last immunization, each buffalo was infected with 1000 Japan cercariae of Schistosoma. Fecal examinations were conducted 2 days and 1 day before the perfusion, and on the day of perfusion. The number of hatching miracidia and eggs per gram feces was recorded. Fifty-six days after the infection, the buffalos were sacrificed and perfused via the descending

  11. 热应激预处理影响鼠背轴型皮瓣HSP70、HSF1表达的实验研究

    Institute of Scientific and Technical Information of China (English)

    刘哲伟; 苏开新; 陈军; 姚建; 李辉; 王海华; 赵鑫

    2014-01-01

    目的探讨热应激预处理(HSP)对缺血皮瓣组织的保护作用及其与皮瓣中热休克蛋白70(HSP70)和热休克因子1(HSF1)的关系。方法将96只雄性Sprague Dawley大鼠随机分为缺血组与实验组(HSP+缺血组)。缺血组:皮瓣模型制备后,阻断皮瓣蒂部第1 h、2 h、4 h、6 h、8 h、10 h共6个时间点,每时间点后观测皮瓣存活面积、HSP70、HSF1的变化,并进行统计学分析;实验组(HSP+缺血组):先予以加热预处理,之后与缺血组相同。结果单纯缺血组皮瓣成活率下降率较实验组(HSP+缺血组)明显,差异有显著性(P〈0.05),热应激预处理各组皮瓣的HSP70、HSF1合成量与皮瓣存活率呈正相关(r=0.7577,P〈0.01)。结论热应激预处理能够改善皮瓣组织的存活率,HSP70、HSF1在热应激预处理改善皮瓣存活率的保护作用中发挥着重要的作用。

  12. 热休克蛋白70过表达对骨骼肌细胞内 ATP水平的影响%Effect of over-expression Hsp70 on the intracellular ATP level in C2C12 cells

    Institute of Scientific and Technical Information of China (English)

    王磊; 王尊; 刘跃飞; 顾一煌

    2013-01-01

    Objective To examine the effect of over-expression Hsp70 on the intracellular ATP level in C2C12 cells. Methods Hsp70 gene was amplified from pAT153 plasmids and then cloned into pTRE2hyg vector. After the transfection of recombinant plasmids of pTRE2hyg-Hsp70 into the C2C12 cells, the expression of Hsp70 was examined by Western blot. Furthermore, the intracellular ATP level was evaluated in C2C12 cells at different time points (0 d, 3 d, 7 d) during cell culture. Results Compared with controls, the intracellular ATP level was significantly increased (P<0.05) at different time points,(14.5 ± 2.87)nmol/mg protein (3 d), (15.3 ± 3.12) nmol/mg protein(7 d) after transfection. Conclusion The C2C12 cells of over-expression Hsp70 can increase the intracellular ATP level, indicating that Hsp70 may play a role in the metabolism in skeletal muscle cells.%目的:探讨热休克蛋白70(heat shock protein 70,Hsp70)过表达对骨骼肌细胞(C2C12)内ATP水平的影响。方法通过构建重组pTRE2hyg-Hsp70质粒,稳定转染C2C12细胞系,建立Hsp70过表达的C2C12细胞系。分别在转染后细胞培养的不同时间点(0 d,3 d,7 d),检测细胞内ATP的水平。结果 Hsp70过表达的C2C12细胞系在培养的3 d,7 d,细胞内ATP的水平分别达到(14.5±2.87)nmol/mg蛋白质、(15.3±3.12)nmol/mg 蛋白质,与对照组相比明显增高(P<0.05)。结论 Hsp70过表达的骨骼肌细胞可以提高细胞内的ATP水平,提示Hsp70对骨骼肌细胞的能量代谢产生影响。

  13. 海豚链球菌感染对不同品系罗非鱼血液生化指标和肝脏HSP70 mRNA表达的影响%Studies on blood biochemical indices and expression of hepatic HSP70 mRNA of different tilapia strains artificially challenged with Streptococcus iniae

    Institute of Scientific and Technical Information of China (English)

    强俊; 杨弘; 王辉; 徐跑; 柒壮林; 何杰

    2012-01-01

    In order to establish the superiority, four different tilapia strains (GIFT tilapia, New GIFT tilapia, Egypt Nile tilapia and Red tilapia) were compared in the study. At day 100 tilapias in the treatment group were intraperitoneally injected with Streptococcus iniae suspension (2.95×108/mL) to evaluate disease resistance of four different tilapia strains against S. iniae in terms of the serum biochemical parameters and hepatic HSP70 mRNA expression before and after infection. Another set of twenty fish taken from each tank was challenged with S. iniae using the same method as above, and the cumulative mortality of 4 strains was recorded at different times post infection. The results showed that GIFT tilapia and New GIFT tilapia were more sensitive to S. iniae at 96 h after infection, cumulative mortalities were 36.67% and 38.33% respectively. Egypt Nile tilapia was less sensitive to pathogen and no death occurred during the experiment. The levels of serum cortisol (COR) and glucose (GLU) and expression levels of HSP70 mRNA in GIFT tilapia, New GIFT tilapia and Red tilapia strains were highly significant(P<0.05), and the activities of glutamic-pyruvic transaminase (ALT), glutamic-oxaloacetic transaminase (AST)and lysozyme (LSZ) also increased, while serum alkaline phosphatase (AKP), triglyceride and cholesterol levels were significantly lower than those prior to infection (P<0.05). By utilizing the energy from resolved hepatin and lipid, the synthesis of hepatic HSP70 mRNA and levels of LSZ and globulin in Egypt Nile tilapia were improved, and thus the non-specific immunity strengthened. It is suggested that in the course of tilapia breeding, effective combination of the disease resistance with growth be conducted so as to guarantee sustainable development of the tilapia industry.%以吉富罗非鱼、新吉富罗非鱼、埃及尼罗罗非鱼和红罗非鱼为研究对象,饲养100 d后,进行海豚链球菌(2.95×108 CFU/mL)感染试验,分析攻毒前后各品

  14. Gymnastics of molecular chaperones.

    Science.gov (United States)

    Mayer, Matthias P

    2010-08-13

    Molecular chaperones assist folding processes and conformational changes in many proteins. In order to do so, they progress through complex conformational cycles themselves. In this review, I discuss the diverse conformational dynamics of the ATP-dependent chaperones of the Hsp60, Hsp70, Hsp90, and Hsp100 families. PMID:20705236

  15. EFFECTS OF ACUTE THERMAL STRESS ON HSP70 MRNA, PHYSIOLOGY AND NONSPECIFIC IMMUNITY IN SIBERIAN STURGEON (ACIPENSER BAERII)%急性热应激对西伯利亚鲟HSP70mRNA表达、血清皮质醇和非特异性免疫的影响

    Institute of Scientific and Technical Information of China (English)

    田照辉; 徐绍刚; 王巍; 胡红霞; 马国庆

    2013-01-01

    The effect of acute thermal stress on HSP1Q mRNA expression, physiology and nonspecific immunity were investigated in Siberian sturgeon. In the experiment, ten fish were transferred froml7.5℃ to 27.5℃ water, then data were measured at lh and 3h. Fish in 17.5℃ water were taken as Oh group. Respiratory rate increased from 80-90 breaths/min to 210 breaths/min, then decreased to 180 breaths/min after 0.5h. Among the three tissues including gill, spleen and brain, the HSP70 mRNA expression in gill rose most quickly after lh stress, and was about 1.63 fold compared with the Oh group (P<0.05), then remained this level to 3h; The HSP10 mRNA expression in spleen and brain had nearly no change in lh thermal stress, then rose quickly from lh to 3h in the two tissues, especially in brain at 3h rose most quickly to about 1.77 fold compared with the lh group (P<0.05). Compared with the Oh group, serum cortisol increased quickly and was 5.14 fold (PO.05) at lh, then decreased quickly to 2.1 fold at 3h. Spleen macrophage respiratory burst achieved the maximum (P<0.05) at lh, then reduced to the Oh group level. Serum complement C3 increased at lh and decreased significantly at 3h (P<0.05). Serum lysozyme activity firstly elevated then decreased but had no significant differences. Serum SOD activity decreased when thermal stress time prolonged, and achieved the minimum at 3h (P<0.05). Serum MDA content decreased when thermal stress time prolonged and had no significant difference. The results showed that lh acute thermal stress promoted the nonspecific immunity and tolerance, but 3h acute thermal stress decreased it.%为研究西伯利亚鲟(Acipenser baerii)对急性热应激的抗逆机理,将体质量为(155.47±19.50) g的鱼从17.5 ℃迅速转至27.5 ℃水中,在1h和3h取样测定HSP70 mRNA表达变化、血清皮质醇和非特异性免疫指标.结果显示:急性热应激时鳃、脾和脑的HSP70 mRNA表达量升高,具有组织特异性,热应激1h时

  16. 逆针灸"关元"穴对更年期大鼠脾脏热休克蛋白70及其mRNA表达和血清白介素-2、肿瘤坏死因子-α含量的影响%Effect of Preventive Acupuncture and Moxibustion at "Guanyuan"(CV 4)on the Expression of HSP 70 and HSP 70 mRNA in Spleen and the Contents of Serum IL-2,TNF-α in Menopausal Rats

    Institute of Scientific and Technical Information of China (English)

    李晓泓; 王洪彬; 徐莉莉; 宋晓琳; 郑玲; 何玉伟; 张露芬

    2009-01-01

    目的:观察逆针灸"关元"穴对自然更年期大鼠免疫功能的影响.方法:经阴道筛查出动情周期紊乱的10月龄SD雌性大鼠160只,随机分为自然10、12、14、16月龄组,逆灸12、14、16月龄组和逆针12、14、16月龄组.另选3.5月龄健康SD雌性大鼠16只为年轻对照组.逆灸、逆针各组从大鼠10月龄开始,每周灸或针"关元"穴2次,共8周.采用免疫组化法检测脾脏热休克蛋白70(HSP 70),原位杂交法检测HSP 70mRNA表达,放射免疫法测定血清白介素-2(IL-2)、肿瘤坏死因子-α(TNF-α)含量.结果:更年期大鼠随着月龄的增长脾脏HSP 70HSP 70 mRNA的表达呈先升后降的变化,血清IL-2含量波动性降低,TNF-α含量先降后升.与自然同月龄组相比,逆灸、逆针各月龄组HSP 70HSP 70 mRNA表达和IL-2含量均呈增加趋势,其中逆灸、逆针12月龄组HSP 70HSP 70 mRNA,逆灸14、16月龄组HSP 70,逆针14月龄组HSP 70 mRNA的表达差异具有统计学意义(P<0.01或P<0.05),IL-2含量逆灸14月龄组明显升高(P<0.05);TNF-α含量逆灸、逆针12月龄组均明显升高(P<0.01),逆灸16月龄组明显降低(P<0.05).同月龄逆灸、逆针组相比,逆针12月龄组HSP 70 mRNA显著升高(P<0.05).结论:逆针灸"关元"穴对调整更年期免疫功能紊乱具有一定作用,其机制可能与调节脾脏组织细胞HSP 70及其基因的表达,启动了机体内在的抗病与应变能力,从而调节血清免疫因子IL-2、TNF-α的含量有关.%Objective To observe the influence of preventive acupuncture (PA) and preventive moxibustion (PM) at "Guanyuan" (CV 4) on the immune function in natural climacteric rats. Methods A total of 160 female SD rats were randomized into control, PA and PM groups, the former one group was further divided into 10 month (mon) ,12 mon, 14 mon and 16 mon subgroups, and the later two groups were further divided into 12 mon, 14 mon and 16 mon subgroups, with 16 rats in each. In addition

  17. Detection and molecular identification protocols for Phyllosticta citricarpa from citrus matter

    Directory of Open Access Journals (Sweden)

    Mariette Truter

    2012-03-01

    Full Text Available Strict quarantine measures for the export of South African citrus fruit to European and US markets require the development of sensitive and accurate detection methods for the pathogen Phyllosticta citricarpa – a fungus causing citrus black spot disease. Because of the presence of other, non-pathogenic Phyllosticta species, rapid and accurate verification of the Phyllosticta species present on exported citrus fruit is important to producers, exporters and regulatory authorities to prevent unnecessary losses. We have analysed over 800 samples collected over 7 years and have compared sample preparation and detection protocols applied in different environments: nurseries, production systems including phytosanitary inspections in orchards, pack houses and export terminals in order to compile protocols for the detection of P. citricarpa. Standard procedures of sample preparation and DNA extraction were adapted to suit diverse inoculum sources. Low pathogen numbers in symptomless green leaves, for example, obliged the use of a wet-dry enrichment technique constituting the stimulation of fungal growth for easier detection. Physical maceration was adapted for sturdy material using liquid nitrogen or bead beating. The use of a two-step polymerase chain reaction (PCR with nested primers significantly increased both the sensitivity and the specificity of the PCR performed on soil samples, overcoming problems with relatively impure DNA extracts and low pathogen numbers. The assays have proven to be highly consistent, thereby providing a reliable, reproducible and highly sensitive detection and diagnostic service to the southern African citrus industries in order to sustain market access.

  18. Molecular characterization of a citrus yellow vein clearing virus strain from China.

    Science.gov (United States)

    Zhen, Song; Kurth, Elizabeth G; Peremyslov, Valera V; Changyong, Zhou; Dolja, Valerian V

    2015-07-01

    The complete nucleotide sequence of an isolate of citrus yellow vein clearing virus from Yunnan, China (CYVCV-RL), was determined following whole-genome amplification by RT-PCR. The CYVCV-RL genome was 7529 nt in length, excluding the 3' poly (A) tail, and contained six open reading frames (ORFs), resembling that of viruses belonging to the genus Mandarivirus in the family Alphaflexiviridae. Sequence analysis showed that the CYVCV-RL shared the greatest nucleotide sequence identity with the CYVCV-Y1 (JX040635) isolate from Turkey for the whole genome (97.1%), 5' UTR (98.7%), 3' UTR (100.0%), and each of six ORFs (96.5% to 97.8%), suggesting that there is apparent genetic stability among CYVCV isolates of different geographic origin. PMID:25913691

  19. A comparative study of survival, metabolism, immune indi-cators and HSP70 expression in three kinds of shell colors Japanese scallop Mizuhopecten yessoensis under high tem-perature stress%高温下3种壳色虾夷扇贝存活率、代谢率、免疫酶活力及HSP70表达的比较研究

    Institute of Scientific and Technical Information of China (English)

    郝振林; 刘京哲; 唐雪娇; 湛垚垚; 田莹; 丁君; 杨立猛; 常亚青

    2015-01-01

    以 3 种壳色虾夷扇贝(Mizuhopecten yessoensis)(枫叶贝、红贝、白贝)为实验材料, 将在 15℃暂养的 3 种壳色虾夷扇贝分别驯化至 20 , 22 , 24 和 26 ℃ 4 个温度梯度(升温幅度为 1 ℃/d), 待各处理组达到对应温度梯度后暂养 7 d, 测定比较 3 种壳色虾夷扇贝的存活率、耗氧率和排氨率、抗氧化酶活力和热激蛋白 HSP70 表达等指标.结果表明: 随着温度的升高, 3 种壳色的虾夷扇贝的存活率呈先上升后下降趋势, 其中, 白壳色虾夷扇贝的存活率在同一处理组中最高; 3 种壳色的虾夷扇贝耗氧和排氨率均随温度升高呈现先升高后降低的趋势, 同一温度处理组下白壳色虾夷扇贝的代谢率始终高于红贝和枫叶贝; 另外, 总抗氧化能力(total-antioxidant capacity, T-AOC)、超氧化物歧酶(activity of superoxide dismutase, SOD)、过氧化氢酶(Catalase, CAT)、过氧化物酶(Peroxidase, POD) 4 种免疫酶的活性随温度升高也呈现先升高后降低的趋势, 且同一温度处理组下白壳色虾夷扇贝的免疫酶活力始终高于红贝和枫叶贝; 3 种壳色虾夷扇贝鳃 HSP70 的表达模式基本相同, 随着温度的逐渐升高, HSP70 持续表达, 到 26℃时表达量最高, 且白壳色扇贝 HSP70 的相对表达量最高为 5.07.综上, 白壳色虾夷扇贝在高温应激下表现出较强的抵抗和耐受能力, 因此, 可将其作为后期耐高温型虾夷扇贝新品系的重要培育材料.%Three shell colors of Mizuhopecten yessoensis were used to test the survival rate, oxygen consumption rate, ammonia-N excretion rate, catalase (CAT) and activity of superoxide dismutase (SOD) activities, to-tal-antioxidant capacity (T-AOC), peroxidase (POD) contents, and HSP70 under short-term high temperature. The scallops reared in 15℃ were acclimatized to the different temperature levels 20, 22, 24 and 26℃ (temperature increased 1℃ per day), and then maintained for 7 days. The

  20. Genetic diversity and phylogenetic analysis of Citrus (L from north-east India as revealed by meiosis, and molecular analysis of internal transcribed spacer region of rDNA

    Directory of Open Access Journals (Sweden)

    Marlykynti Hynniewta

    2014-12-01

    Full Text Available The north-eastern region of India is reported to be the center of origin and rich in diversity of Citrus (L. species, where some wild and endangered species namely Citrus indica, Citrus macroptera, Citrus latipes, Citrus ichagensis and Citrus assamensis exist in their natural and undisturbed habitat. In order to have comprehensive information about the extent of genetic variability and the occurrence of cryptic genomic hybridity between and within various Citrus species, a combined approach involving morphological, cytogenetical and molecular approaches were adopted in the present study. Cytogenetic approaches are known to resolve taxonomic riddles in a more efficient manner, by clearly delineating taxa at species and sub species levels. Male meiotic studies revealed a gametic chromosome number of n = 9, without any evidence of numerical variations. Bivalents outnumbered all other types of associations in pollen mother cells (PMCs analyzed at diplotene, diakinesis and metaphase I. Univalents were frequently encountered in nine species presently studied, though their presence appropriately did not influence the distributional pattern of the chromosomes at anaphases I and II. The molecular approaches for phylogenetic analysis based on sequence data related to ITS 1, ITS 2 and ITS 1 + 5.8 s + ITS 2 of rDNA using maximum parsimony method and Bayesian inference have thrown light on species inter-relationship and evolution of Citrus species confirming our cytogenetical interpretations. The three true basic species i.e. Citrus medica, Citrus maxima and Citrus reticulata with their unique status have been resolved into distinct clades with molecular approaches as well. C. indica which occupies a unique position in the phylogenetic ladder of the genus Citrus has been resolved as a distinct clade and almost behaving as an out-group. The presences of quadrivalents in C. indica also echo and support its unique position. From our study it is amply

  1. A water-soluble high molecular weight substance isolated from Hyuganatsu orange (Citrus tamurana), suspected to be a polysaccharide, inhibits rat osteoclast cell formation

    OpenAIRE

    Hiroko Hata; Masatoshi Yamaguchi; Hiroshi Sameshima; Tsuyomu Ikenoue; Junko Matsubara; Makoto Tsuboi; Takashi Tanaka

    2015-01-01

    Background: Osteoporosis is detrimental to aged women’s health care. We previously reported that Hyuganatsu orange (Citrus Tamurana) contains active substances that inhibit osteoclast activities. Prior to conducting a human study, we sought to identify the biological active substance in the Hyuganatsu orange which suppresses osteoclast formation. Methods: We isolated five fractions from a Hyuganatsu orange extract according to molecular weight. Each fraction was tested to determine its...

  2. Metabolic fates and effects of nitrite in brown trout under normoxic and hypoxic conditions: blood and tissue nitrite metabolism and interactions with branchial NOS, Na+/K+-ATPase and hsp70 expression.

    Science.gov (United States)

    Jensen, Frank B; Gerber, Lucie; Hansen, Marie N; Madsen, Steffen S

    2015-07-01

    Nitrite secures essential nitric oxide (NO) bioavailability in hypoxia at low endogenous concentrations, whereas it becomes toxic at high concentrations. We exposed brown trout to normoxic and hypoxic water in the absence and presence of added ambient nitrite to decipher the cellular metabolism and effects of nitrite at basal and elevated concentrations under different oxygen regimes. We also tested hypotheses concerning the influence of nitrite on branchial nitric oxide synthase (NOS), Na(+)/K(+)-ATPase (nka) and heat shock protein (hsp70) mRNA expression. Basal plasma and erythrocyte nitrite levels were higher in hypoxia than normoxia, suggesting increased NOS activity. Nitrite exposure strongly elevated nitrite concentrations in plasma, erythrocytes, heart tissue and white muscle, which was associated with an extensive metabolism of nitrite to nitrate and to iron-nitrosylated and S-nitrosated compounds. Nitrite uptake was slightly higher in hypoxia than normoxia, and high internal nitrite levels extensively converted blood hemoglobin to methemoglobin and nitrosylhemoglobin. Hypoxia increased inducible NOS (iNOS) mRNA levels in the gills, which was overruled by a strong inhibition of iNOS expression by nitrite in both normoxia and hypoxia, suggesting negative-feedback regulation of iNOS gene expression by nitrite. A similar inhibition was absent for neuronal NOS. Branchial NKA activity stayed unchanged, but mRNA levels of the nkaα1a subunit increased with hypoxia and nitrite, which may have countered an initial NKA inhibition. Nitrite also increased hsp70 gene expression, probably contributing to the cytoprotective effects of nitrite at low concentrations. Nitrite displays a concentration-dependent switch between positive and negative effects similar to other signaling molecules. PMID:25908056

  3. Physiological and Molecular Responses to Excess Boron in Citrus macrophylla W

    Science.gov (United States)

    Martínez-Cuenca, Mary-Rus; Martínez-Alcántara, Belén; Quiñones, Ana; Ruiz, Marta; Iglesias, Domingo J.; Primo-Millo, Eduardo; Forner-Giner, M. Ángeles

    2015-01-01

    This work provides insight into several mechanisms involved in boron (B) regulation pathway in response to high B conditions in Citrus. The study was carried out in Citrus macrophylla W. (Cm) seedlings cultured “in vitro” in media with 50 or 400 μM H3BO3 (control, Ct, and B-excess, +B, plants, respectively). Growth parameters, B concentration, leaf chlorophyll (Chl) concentration, the expression of the main putative genes involved in B transport and distribution, and leaf and root proline and malonaldehyde (MDA) concentrations, were assessed. Excess B led to high B concentration in +B plants (3.8- and 1.4-fold in leaves and roots, respectively) when compared with Ct ones. However, a minor effect was recorded in the plant (incipient visual symptoms, less than 27% reduction in root growth and 26% decrease in Chl b concentration). B toxicity down-regulated by half the expression level of putative B transporter genes NIP5 and PIP1. CmBOR1 gene was not repressed in +B plants and B accumulated in the shoots. High B level increased the transcripts of putative gene TIP5, involved in B transport across the tonoplast, by 3.3- and 2.4-fold in leaves and roots, respectively. The activity of V-PPiase proton pump, related with the electrochemical gradient in the vacuole, was also enhanced in +B organs. B toxicity up-regulated putative BOR4 gene (2.1- and 2.7-fold in roots and leaves, respectively), which codifies for an active efflux B transporter. Accordingly, B was located in +B plants preferently in an insoluble form on cell walls. Finally, excess B caused a significant rise in proline concentration (51% and 34% in roots and leaves, respectively), while the MDA level did not exceed 20%. In conclusion, Cm tolerance to a high B level is likely based on the synergism of several specific mechanisms against B toxicity, including: 1/ down-regulation of NIP5 and PIP1 boron transporters; 2/ activation of B efflux from cells due to the up-regulation of putative BOR4 gene; 3

  4. Physiological and Molecular Responses to Excess Boron in Citrus macrophylla W.

    Directory of Open Access Journals (Sweden)

    Mary-Rus Martínez-Cuenca

    Full Text Available This work provides insight into several mechanisms involved in boron (B regulation pathway in response to high B conditions in Citrus. The study was carried out in Citrus macrophylla W. (Cm seedlings cultured "in vitro" in media with 50 or 400 μM H3BO3 (control, Ct, and B-excess, +B, plants, respectively. Growth parameters, B concentration, leaf chlorophyll (Chl concentration, the expression of the main putative genes involved in B transport and distribution, and leaf and root proline and malonaldehyde (MDA concentrations, were assessed. Excess B led to high B concentration in +B plants (3.8- and 1.4-fold in leaves and roots, respectively when compared with Ct ones. However, a minor effect was recorded in the plant (incipient visual symptoms, less than 27% reduction in root growth and 26% decrease in Chl b concentration. B toxicity down-regulated by half the expression level of putative B transporter genes NIP5 and PIP1. CmBOR1 gene was not repressed in +B plants and B accumulated in the shoots. High B level increased the transcripts of putative gene TIP5, involved in B transport across the tonoplast, by 3.3- and 2.4-fold in leaves and roots, respectively. The activity of V-PPiase proton pump, related with the electrochemical gradient in the vacuole, was also enhanced in +B organs. B toxicity up-regulated putative BOR4 gene (2.1- and 2.7-fold in roots and leaves, respectively, which codifies for an active efflux B transporter. Accordingly, B was located in +B plants preferently in an insoluble form on cell walls. Finally, excess B caused a significant rise in proline concentration (51% and 34% in roots and leaves, respectively, while the MDA level did not exceed 20%. In conclusion, Cm tolerance to a high B level is likely based on the synergism of several specific mechanisms against B toxicity, including: 1/ down-regulation of NIP5 and PIP1 boron transporters; 2/ activation of B efflux from cells due to the up-regulation of putative BOR4 gene

  5. Molecular characterization of two carboxylesterase genes of the citrus red mite, Panonychus citri (Acari: Tetranychidae).

    Science.gov (United States)

    Zhang, Kun; Niu, Jin-Zhi; Ding, Tian-Bo; Dou, Wei; Wang, Jin-Jun

    2013-04-01

    The citrus red mite, Panonychus citri, is known for its ability to rapidly evolve resistance to insecticides/acaricides and to adapt to hosts that produce toxins. To get better insight into the detoxification mechanism of P. citri, two carboxylesterase (CarE) genes, PCE1 and PCE2, were isolated and characterized. PCE1 and PCE2 contained open reading frames of 1,653 and 1,392 nucleotides, encoding proteins of 550 and 463 amino acid residues, respectively. Phylogenetic analyses showed that PCE1 and PCE2 were most closely related to the CarE genes from other phytophagous mites. The transcriptional profiles of two CarE genes among developmental stages (egg, larva, nymph, adult female, and adult male), after exposing to four acaricides (avermectin, azocyclotin, pyridaben, and spirodiclofen) and acid rain were investigated using real-time quantitative PCR (qPCR). The results showed that during development, PCE1 was highly expressed at the egg stage, whereas PCE2 was abundantly expressed at the adult stage of males. The expression levels of PCE1 were highly induced upon exposure to acaricides and acid rain. On the other hand, the expression levels of PCE2 were increased after treatment with avermectin and pyridaben. These results suggest that PCE1 and PCE2 may have distinct roles in different developmental stages and participate in the detoxification of acaricides. PMID:23404785

  6. Hsp70/C D80 DNA vaccine inhibits asthma by regulating the balance of Th1/Th2/Treg/Th1 7 in an acute mouse model%HSP70/C D80 DNA疫苗通过调节Th1/Th2/Treg/Th17细胞对小鼠急性哮喘的抑制作用

    Institute of Scientific and Technical Information of China (English)

    李燕; 谢敏; 史小玲; 王晓燕; 唐利; 钟森; 陈庄

    2014-01-01

    Objective To explore the mechanism of Th1/Th2/Treg/Th17 rebalance in an acute asthma mouse model treated with heat shock protein70 (Hsp70)/C D80 DNA vaccine.Methods Forty BALB/c mice were divided into the control group,asthma-model group,pVXA1-vector group and vaccine treatment group,10 in each group.They were intraperitoneally injected into ovalbumin (OVA)to build acute asthma model and treated with DNA vaccine.The air-way responsiveness was evaluated after inhaling the methacholine (Mch).The IgE content in serum was detected by ELISA.The pathological change of lung tissue was observed by HE and AB-PAS staining.The expressions of interferon-γ(IFN-γ),interleukin-4 (IL-4),ransforming growth factor beta (TGF-β)and interleukin-17 (IL-17)in BALF were measured by ELISA.The expressions of T-bet,GATA binding protein 3 (GATA-3 ),forkheah box protein3 (Foxp3 ) and retinoid related orphan receptor gamma t (RORγt)in lung tissue were detected by Real-time PCR.Results Com-pared with asthmatic mouse,mouse treated with vaccine exhibited the decreased airway responsiveness,IgE (all P<0.05)and inflammatory infiltration of lung tissues,repressed proliferation of goblet cells,and increased IFN-γ/IL-4, TGF-β/IL-17,T-bet/GATA-3 and Foxp3/RORγt (all P<0.05).Conclusion The Hsp70/C D80 DNA vaccine plays a therapeutic role in asthma by restoring the balance of Th1/Th2/Treg/Th17 cell.%目的:考察热休克蛋白70(HSP70)/C D80 DNA疫苗治疗小鼠急性哮喘的疗效,研究该过程中Th1/Th2/Treg/Th17再平衡的作用机制。方法 BALB/c小鼠40只随机分为空白对照组(空白组),哮喘模型组(模型组), pVAX1(+)空载体对照组(空载组)和 HSP70/C D80疫苗治疗组(治疗组),每组10只。鸡卵清蛋白(OVA)激发致敏小鼠构建急性哮喘模型,HSP70/C D80 DNA疫苗进行治疗。吸入性支气管激发试验测定气道反应性,ELISA法测定血清中IgE的含量,HE和AB-PAS染色观察肺组织病理学

  7. The destructive citrus pathogen, 'Candidatus Liberibacter asiaticus' encodes a functional flagellin characteristic of a pathogen-associated molecular pattern.

    Directory of Open Access Journals (Sweden)

    Huasong Zou

    Full Text Available Huanglongbing (HLB is presently the most devastating citrus disease worldwide. As an intracellular plant pathogen and insect symbiont, the HLB bacterium, 'Candidatus Liberibacter asiaticus' (Las, retains the entire flagellum-encoding gene cluster in its significantly reduced genome. Las encodes a flagellin and hook-associated protein (Fla of 452 amino acids that contains a conserved 22 amino acid domain (flg22 at positions 29 to 50 in the N-terminus. The phenotypic alteration in motility of a Sinorhizobium meliloti mutant lacking the fla genes was partially restored by constitutive expression of Fla(Las. Agrobacterium-mediated transient expression in planta revealed that Fla(Las induced cell death and callose deposition in Nicotiana benthamiana, and that the transcription of BAK1 and SGT1, which are associated with plant innate immunity, was upregulated. Amino acid substitution experiments revealed that residues 38 (serine and 39 (aspartate of Fla(Las were essential for callose induction. The synthetic flg22(Las peptide could not induce plant cell death but retained the ability to induce callose deposition at a concentration of 20 µM or above. This demonstrated that the pathogen-associated molecular pattern (PAMP activity of flg22 in Las was weaker than those in other well-studied plant pathogenic bacteria. These results indicate that Fla(Las acts as a PAMP and may play an important role in triggering host plant resistance to the HLB bacteria.

  8. 脑损伤后热休克蛋白70及热休克因子Ⅰ基因转录的实验研究%Experimental research of the transcription of HSP70and HSF1 gene after traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    赵连旭; 徐小虎; 潘速跃; 张成

    2002-01-01

    目的为了解脑损伤后应激基因转录的状况.方法采用RT-PCR对伤后脑组织HSP70、HSF1 mRNA的数量进行测定.结果在脑损伤15 min后脑干HSP70、HSF1 mRNA水平显著升高,在损伤后迅速死亡的致死损伤组的鼠脑中HSP70、HSF1 mRNA水平亦显著升高.结论应激基因的转录水平在脑损伤后明显提高,脑应激蛋白合成增强,参与损伤与修复过程.同时由于HSP70、HSF1 mRNA对脑损伤的迅速反应能力,使之成为观察脑损伤的灵敏指标.

  9. 苯并芘处理对人支气管上皮细胞热休克蛋白70和着色性干皮病G蛋白表达的影响%Changes of Hsp70 and XPG in human bronchial epithelia cells treated with benzo[a] prrene

    Institute of Scientific and Technical Information of China (English)

    杨瑾; 刘晓勇; 邹云锋; 牛丕业; 段燕英; 邬堂春

    2010-01-01

    目的 探讨苯并芘(Benzo[a]pyrene,BaP)作用下的人支气管上皮细胞(16HBE)热休克蛋白70(Heat shock protein 70,Hsp70)和着色性干皮病G蛋白(Xeroderma pigmentosum group G,XPG)表达的时间效应特征,并分析二者之间可能存在的关联性.方法 用8 μmol/L BaP处理16HBE细胞0、1、2、4、8、12、24和48 h,以彗星实验检测细胞DNA损伤并以Olive尾矩值(Olive Tail Moments,OTM)评价DNA损伤程度,以Western-blot检测Hsp70和XPG的表达水平,并以激光共聚焦法检测二者的共定位.结果 染毒1~2h时OTM值变化率最大,与正常细胞相比,除染毒1h组外其余各组OTM值均显著性增高(P<0.01);Hsp70和XPG的表达随染毒时间逐渐增高,12 h时达到峰值,激光共聚焦结果显示BaP染毒细胞后Hsp70和XPG的结合在细胞核内增强.结论 在该试验条件下,XPG在执行核苷酸切除修复作用时可能有Hsp70的参与,其具体机制还有待其他实验进一步加以证实.

  10. La proteína de estrés calórico Hsp70 funciona como un indicador de adaptación de los bovinos a las zonas áridas

    Directory of Open Access Journals (Sweden)

    Bañuelos-Valenzuela, Rómulo, Sánchez-Rodríguez, Sergio H.

    2005-03-01

    Full Text Available El propósito del presente estudio fue seleccionar genotipos de ganado bovino que estén mas adaptados al estrés calórico en las zonas áridas, a través de la expresión de las proteínas de choque calórico (Hsp, las cuales, son constituyentes normales de las células y su síntesis se incrementa con la exposición a varias formas de estrés. Nuestro objetivo fue establecer, si la expresión de las Hsp de los linfocitos del ganado bovino explotado en la región, muestra diferencias raciales en respuesta al estés calórico. De los bovinos, se obtuvieron linfocitos de sangre periférica por gradientes de Ficoll-histopaque, después se cultivaron en medio RPMI a temperaturas de 38, 40, 42 y 44°C, durante 4 horas, posteriormente se lisaron, se obtuvieron extractos que se caracterizaron por PAGE-SDS 10%, se transfirieron a papel de nitrocelulosa y se probaron con anticuerpos monoclonales anti-Hsp 25, 60,70 y 90. Se encontró: El calor es un agente estresante en los linfocitos del ganado bovino explotado en la región, cuya respuesta es a través de la expresión de la proteína Hsp70. La cantidad de proteína Hsp70 expresada en los linfocitos esta en función al estrés calórico recibido. El nivel de adaptación de un organismo ante un estresor, esta en función a la cantidad de Hsp expresada, por lo tanto, las razas bovinas más adaptadas a la región en orden decreciente es: Holstein-Australiano, Criollo, Pardo Suizo, Holstein y Limousin.

  11. Citrus Genomics

    OpenAIRE

    Talon, Manuel; Gmitter, Fred G.Jr.

    2008-01-01

    Citrus is one of the most widespread fruit crops globally, with great economic and health value. It is among the most difficult plants to improve through traditional breeding approaches. Currently, there is risk of devastation by diseases threatening to limit production and future availability to the human population. As technologies rapidly advance in genomic science, they are quickly adapted to address the biological challenges of the citrus plant system and the world's industries. The hist...

  12. Efeito do estresse térmico agudo sobre os níveis da proteína e RNA mensageiro da Hsp70, em fígado e cérebro de pintos de corte de diferentes linhagens Effect of acute heat stress on hepatic and cerebral messenger RNA heat shock protein 70 and heat shock protein 70 level of broiler chicks from 2 to 5 days old of different strains

    Directory of Open Access Journals (Sweden)

    Nelson José Laurino Dionello

    2001-10-01

    Full Text Available Oitenta pintos de corte de duas linhagens, pescoço pelado (Na/na e Hubbard-Pettersen, entre 2 e 5 dias de idade, foram expostos a estresse térmico agudo (36-37°C, durante cinco horas. Foram avaliados temperatura cloacal e peso corporal individuais ao início e final do período de estresse térmico agudo. Os pintos foram sacrificados ao final e amostras de fígado e cérebro foram coletadas e analisadas por Western Blotting e Northern Blotting, para quantificação da proteína e RNA mensageiro da Hsp70, respectivamente. Os resultados apresentaram maiores níveis protéicos de Hsp70 no tecido cerebral do que no hepático. Antes do estresse térmico, não houve diferenças de linhagens para expressão e síntese de Hsp70, em ambos os tecidos. Após o estresse térmico, as aves estressadas apresentaram maiores níveis protéicos de Hsp70, quando o tecido analisado foi o fígado (para as pescoço pelado nas idades de 4 e 5 dias, e menores níveis, em relação às controles, quando o tecido analisado foi o cérebro (para as Pescoço pelado na idade de 2 dias. O tamanho do transcrito de RNA mensageiro de Hsp70 foi de 2,7 kb. Os resultados do presente experimento sugerem que, para o tecido cerebral, a indução de Hsp70 ocorreu em níveis transcripcional e traducional e para o tecido hepático foi detectada apenas alteração em nível traducional, para ambas as linhagens.Eighty broiler chicks, Naked neck (Na/na and Hubbard-Pettersen strains, were exposed to acute heat stress (36-37°C, from 2 to 5 days of age, and body weight and cloacal temperature were measured in the beginning and at the end of the heat stress period (5 h. Birds were sacrificed at the end and liver and brain samples were collected and analyzed using Western Blotting and Northern Blotting to quantify Hsp70 levels and mRNA Hsp70 transcript, respectively. The brain tissue had higher Hsp70 level than liver tissue. Before heat stress it was not observed difference in the Hsp70

  13. Molecular characterization of vitellogenin and its receptor genes from citrus red mite, Panonychus citri (McGregor).

    Science.gov (United States)

    Zhong, Rui; Ding, Tian-Bo; Niu, Jin-Zhi; Xia, Wen-Kai; Liao, Chong-Yu; Dou, Wei; Wang, Jin-Jun

    2015-01-01

    The production and uptake of yolk protein play an important role in the reproduction of all oviparous organisms. Vitellogenin (Vg) is the precursor of vitellin (Vn), which is the major egg storage protein, and vitellogenin receptor (VgR) is a necessary protein for the uptake of Vg into developing oocytes. In this paper, we characterize the full-length Vg and VgR, PcVg1 and PcVgR, respectively, of the citrus red mite Panonychus citri (McGregor). The PcVg1 cDNA is 5748 nucleotides (nt) with a 5553-nt open reading frame (ORF) coding for 1851 amino acids (aa), and the PcVgR is 6090 nt, containing an intact ORF of 5673 nt coding an expected protein of 1891 aa. The PcVg1 aa sequence shows a typical GLCG domain and several K/RXXR cleavage sites, and PcVgR comprises two ligand-binding domains, two epidermal growth factor (EGF)-like regions containing YWTD motifs, a transmembrane domain, and a cytoplasmic domain. An analysis of the aa sequences and phylogenetics implied that both genes were genetically distinct from those of ticks and insects. The transcriptional profiles determined by real-time quantitative PCR in different developmental stages showed that both genes present the same expressional tendencies in eggs, larvae, nymphs, and adults. This suggested that the biosynthesis and uptake of PcVg occurs coordinately. The strong reproductive capacity of P. citri has been hypothesized as an important factor in its resistance; consequently, understanding the molecular mechanisms regulating Vg and VgR are fundamental for mite control. PMID:25739087

  14. Molecular Characterization of Vitellogenin and Its Receptor Genes from Citrus Red Mite, Panonychus citri (McGregor

    Directory of Open Access Journals (Sweden)

    Rui Zhong

    2015-03-01

    Full Text Available The production and uptake of yolk protein play an important role in the reproduction of all oviparous organisms. Vitellogenin (Vg is the precursor of vitellin (Vn, which is the major egg storage protein, and vitellogenin receptor (VgR is a necessary protein for the uptake of Vg into developing oocytes. In this paper, we characterize the full-length Vg and VgR, PcVg1 and PcVgR, respectively, of the citrus red mite Panonychus citri (McGregor. The PcVg1 cDNA is 5748 nucleotides (nt with a 5553-nt open reading frame (ORF coding for 1851 amino acids (aa, and the PcVgR is 6090 nt, containing an intact ORF of 5673 nt coding an expected protein of 1891 aa. The PcVg1 aa sequence shows a typical GLCG domain and several K/RXXR cleavage sites, and PcVgR comprises two ligand-binding domains, two epidermal growth factor (EGF-like regions containing YWTD motifs, a transmembrane domain, and a cytoplasmic domain. An analysis of the aa sequences and phylogenetics implied that both genes were genetically distinct from those of ticks and insects. The transcriptional profiles determined by real-time quantitative PCR in different developmental stages showed that both genes present the same expressional tendencies in eggs, larvae, nymphs, and adults. This suggested that the biosynthesis and uptake of PcVg occurs coordinately. The strong reproductive capacity of P. citri has been hypothesized as an important factor in its resistance; consequently, understanding the molecular mechanisms regulating Vg and VgR are fundamental for mite control.

  15. Molecular Cloning and Identification of a Heat Shock Cognate Protein 70 Gene, Thhsc70, in Thellungiella halophila

    Institute of Scientific and Technical Information of China (English)

    ZHANGXia; GUOShan-Li; YINHai-Bo; XIONGDong-Jin; ZHANGHui; ZHAOYan-Xiu

    2004-01-01

    Heat shock cognate proteins 70 (hsp70s) act as molecular chaperones. Some hsp70s are also expressed in unstressed plants, known as hsc70. To gain further knowledge about the hsc70, the Thellungiella halophila hsc70 (Thhsc70) gene that encoded the cytosolic hsc70 in salt cress (T.halophila (C.A.Mey.) O.E.Schulz) was identified. In unstressed plants the expression of Thhsc70was shown to be tissue-specific. The Thhsc70 gene was induced by heat and cold stresses, but almost not by salt and drought stresses. Overexpression of Thhsc7Ocould increase thermctolerance and chilling tolerance in transgenic Arabidopsis plants.

  16. 加味七方胃痛颗粒对慢性萎缩性胃炎大鼠胃蛋白酶含量及HSP70蛋白表达的影响%Effect of Modified Qifang Weitong Granules on Pepsin Content and Heat Shock Protein 70 Expression in Rats with Chronic Atrophic Gastritis

    Institute of Scientific and Technical Information of China (English)

    唐梅文; 王伟; 李桂贤; 岳小红; 陈国忠; 邓嫦

    2011-01-01

    Objective To investigate the effects of Modified Qifan Weitong Granules(QWG) on pepsin content and heat shock protein 70(HSP70) expression in the gastric mucosa of rats with chronic atrophic gastritis(CAG).Methods CAG rat models were established by means of N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), and the effect of QWG on relative indexes was observed and compared with that of Weifuchun Granules. Results Compared with the model group, the atrophy and intestinal metaplasia of gastric mucosa were relieved in large-and medium-dosage QWG groups and in Weifuchun Granules group, and the pepsin content and HSP70 expression were increased in large-, medium-and small-dosage QWG groups and in Weifuchun Granules group, the difference being significant(P< 0.01). Compared with Weifuchun Granules group, pepsin content and HSP70 expression in large-dosage QWG group were increased with statistical significance(P < 0.05). Conclusion The mechanism of QWG in preventing and treating CAG may be related with prevention of gastric mucosa through increasing pepsin con tent and HSP70 expression.%目的 探讨加味七方胃痛颗粒对慢性萎缩性胃炎(CAC)模型大鼠胃黏膜组织中胃蛋白酶含量及热休克蛋白70(HSP70)表达的影响.方法 采用化学致癌物N-甲基-N'-硝基-N-亚硝基胍(MNNG)建立CAG动物模型,以胃复春为对照组,观察加味七方胃痛颗粒对大鼠CAG相关指标的作用.结果 与模型组比较,加味七方胃痛颗粒高、中剂量组和胃复春组的胃黏膜萎缩程度、胃黏膜肠化生程度明显减轻(P<0.01);加味七方胃痛颗粒高、中、低剂量组和胃复春组胃蛋白酶的含量、HSP70蛋白表达显著增加(P<0.01);与胃复春组比较,加味七方胃痛颗粒高剂量组胃蛋白酶的含量、HSP70蛋白表达显著增加(P<0.05).结论 加味七方胃痛颗粒防治CAG的作用机制可能是通过升高胃蛋白酶的含量及促进HSP70的表达,从而发挥对黏膜保护作用.

  17. 艾灸对亚急性衰老大鼠松果体细胞数及热休克蛋白70表达的影响%Effect of Moxibustion on Quantity of Pinealocytes and Pineal HSP 70 Expression in Subacute Aging Rats

    Institute of Scientific and Technical Information of China (English)

    梁欣; 钟愉

    2011-01-01

    Objective To study the mechanism of moxibustion in postponing the process of aging. Methods Thirty Wistar rats were equally randomized into control, model and moxibustion groups. The subacute aging model was established by hypodermic injection of 25% D-galactose (125 mg/kg). Moxibustion was applied to bilateral "Shenshu" (GV23) and "Pishu" (GV 20) once everyday for 6 weeks. After slicing, the pineal gland tissue was stained with HE and insitu hybridization methods respectively for detecting the quantity of pinealocytes and the expression of heat shock protein (HSP 70).Results Compared with the control group, both the quantity of pinealocytes and the expression of HSP 70 in the pineal gland in the model group were downregulated significantly (P<0. 001, P<0. 01). Compared with model group, the quantity of pinealocytes and of HSP 70 mRNA in the pineal gland of moxibustion group were upregulated significantly( P<0. 01, P<0. 05). Conclusion Moxibustion can suppress aging induced decrease of pinealocyte number and HSP 70 expression in subacute aging rats, which may contribute to its effect in postponing aging.%目的:探讨艾灸延缓衰老的可能作用机制.方法:将30只实验大鼠随机分为空白组、模型组、艾灸组,每组10只.皮下注射25%D-半乳糖溶液(125 mg/kg)造成大鼠衰老模型.艾灸组选取双侧"脾俞""肾俞"进行艾灸治疗,每日治疗1次,每次每穴灸5 min,连续治疗6周.采用组织化学染色及多克隆杭体免疫组化技术,结合细胞医学图像分析及病理图文图像分析方法,测定各组大鼠松果体细胞数及HSP 70表达.结果:与空白组相比,模型组大鼠松果体细胞数显著减少(P<0.001),HSP 70表达显著降低(P<0.01).与模型组相比,艾灸组大鼠松果体细胞数显著增多(P<0.01),HSP 70表达显著增加(P<0.05).结论:艾灸可能通过上调脑区松果体中HSP 70表达、减缓机体衰老过程中松果体细胞减少的速度,在一定程度上维持松果体的正

  18. Influence of In Vitro IL-2 or IL-15 Alone or in Combination with Hsp 70 Derived 14-Mer Peptide (TKD on the Expression of NK Cell Activatory and Inhibitory Receptors on Peripheral Blood T Cells, B Cells and NKT Cells.

    Directory of Open Access Journals (Sweden)

    Ilona Hromadnikova

    Full Text Available Previous studies from Multhoff and colleagues reported that plasma membrane Hsp70 acts as a tumour-specific recognition structure for activated NK cells, and that the incubation of NK cells with Hsp70 and/or a 14-mer peptide derived from the N-terminal sequence of Hsp70 (TKDNNLLGRFELSG, TKD, aa 450-463 plus a low dose of IL-2 triggers NK cell proliferation and migration, and their capacity to kill cancer cells expressing membrane Hsp70. Herein, we have used flow cytometry to determine the influence of in vitro stimulation of peripheral blood mononuclear cells from healthy individuals with IL-2 or IL-15, either alone or in combination with TKD peptide on the cell surface expression of CD94, NK cell activatory receptors (CD16, NK2D, NKG2C, NKp30, NKp44, NKp46, NKp80, KIR2DL4, DNAM-1 and LAMP1 and NK cell inhibitory receptors (NKG2A, KIR2DL2/L3, LIR1/ILT-2 and NKR-P1A by CD3+CD56+ (NKT, CD3+CD4+, CD3+CD8+ and CD19+ populations. NKG2D, DNAM-1, LAMP1 and NKR-P1A expression was upregulated after the stimulation with IL-2 or IL-15 alone or in combination with TKD in NKT, CD8+ T cells and B cells. CD94 was upregulated in NKT and CD8+ T cells. Concurrently, an increase in a number of CD8+ T cells expressing LIR1/ILT-2 and CD4+ T cells positive for NKR-P1A was observed. The proportion of CD8+ T cells that expressed NKG2D was higher after IL-2/TKD treatment, when compared with IL-2 treatment alone. In comparison with IL-15 alone, IL-15/TKD treatment increased the proportion of NKT cells that were positive for CD94, LAMP1 and NKRP-1A. The more potent effect of IL-15/TKD on cell surface expression of NKG2D, LIR1/ILT-2 and NKRP-1A was observed in B cells compared with IL-15 alone. However, this increase was not of statistical significance. IL-2/TKD induced significant upregulation of LAMP1 in CD8+ T cells compared with IL-2 alone. Besides NK cells, other immunocompetent cells present within the fraction of peripheral blood mononuclear cells were influenced by

  19. Evaluation of molecular chaperons Hsp72 and neuropeptide Y as characteristic markers of adaptogenic activity of plant extracts.

    Science.gov (United States)

    Asea, Alexzander; Kaur, Punit; Panossian, Alexander; Wikman, Karl Georg

    2013-11-15

    We have previously demonstrated that ADAPT-232, a fixed combination of adaptogenic substances derived from Eleutherococcus senticosus root extract, Schisandra chinensis berry extract, Rhodiola rosea root extract stimulated the expression and release of neuropeptide Y (NPY) and molecular chaperone Hsp72 from isolated human neurolgia cells. Both of these mediators of stress response are known to play an important role in regulation of neuroendocrine system and immune response. We further demonstrated that ADAPT-232 induced release of Hsp70 is mediated by NPY, suggesting an existence of NPY-mediated pathway of activation of Hsp72 release into the blood circulation system. The objective of this study was to determine whether this pathway is common for adaptogens and whether NPY and/or Hsp72 can be considered as necessary specific biomarkers for adaptogenic activity. The release of NPY and Hsp72 from neuroglia cells in response to treatment with various plant extracts (n=23) including selected validated adaptogens, partly validated adaptogens, claimed but negligibly validated adaptogens and some other plant extracts affecting neuroendocrine and immune systems but never considered as adaptogens was measured using high throughput ELISA techniques. We demonstrated that adaptogens, e.g. R. rosea, S. chinensis and E. senticosus stimulate both NPY and Hsp70 release from neuroblastoma cells, while tonics and stimulants have no significant effect on NPY in this in vitro test. In the groups of partly validated adaptogens the effect of Panax ginseng and Withania somnifera was not statistically significant both on NPY and Hsp70 release, while the activating effect of Bryonia alba and Rhaponticum cartamoides was significant only on Hsp70. In contrast, all tested non-adaptogens, such as antiinflammatoty plant extracts Matricaria recutita, Pelargonium sidoides, Hedera helix and Vitis vinifera significantly inhibit Hsp70 release and have no influence on NPY release from neuroblastoma

  20. A water-soluble high molecular weight substance isolated from Hyuganatsu orange (Citrus tamurana, suspected to be a polysaccharide, inhibits rat osteoclast cell formation

    Directory of Open Access Journals (Sweden)

    Hiroko Hata

    2015-06-01

    Full Text Available Background: Osteoporosis is detrimental to aged women’s health care. We previously reported that Hyuganatsu orange (Citrus Tamurana contains active substances that inhibit osteoclast activities. Prior to conducting a human study, we sought to identify the biological active substance in the Hyuganatsu orange which suppresses osteoclast formation. Methods: We isolated five fractions from a Hyuganatsu orange extract according to molecular weight. Each fraction was tested to determine its suppressive effect on the formation of osteoclasts in rats. We also used high-performance liquid chromatography (HPLC, infra-red (IR, and 1H and 13C NMR spectroscopy to evaluate its chemical structure. Data was recorded as mean ± standard error of the mean. The Mann-Whitney test was used, and a p-value of <.05 was considered statistically significant. Results: The highest and lowest molecular weight fractions showed significant suppression activity on rat osteoclast formation (p < .05. The lowest molecular weight fraction was identified as hesperidin using thin layer chromatography. Additionally, IR absorption revealed that the highest molecular weight fraction was not a flavonoid. With regard to chemical structure, 1H and 13C NMR spectroscopy suggested that the highest molecular weight fraction had signals compatible with a polysaccharide such as galacturonic acid. Conclusions: Hyuganatsu orange contains a biological active substance other than hesperidin that may be a polysaccharide and may suppress osteoclast formation.

  1. Molecular Characterization of Vitellogenin and Its Receptor Genes from Citrus Red Mite, Panonychus citri (McGregor)

    OpenAIRE

    Rui Zhong; Tian-Bo Ding; Jin-Zhi Niu; Wen-Kai Xia; Chong-Yu Liao; Wei Dou; Jin-Jun Wang

    2015-01-01

    The production and uptake of yolk protein play an important role in the reproduction of all oviparous organisms. Vitellogenin (Vg) is the precursor of vitellin (Vn), which is the major egg storage protein, and vitellogenin receptor (VgR) is a necessary protein for the uptake of Vg into developing oocytes. In this paper, we characterize the full-length Vg and VgR, PcVg1 and PcVgR, respectively, of the citrus red mite Panonychus citri (McGregor). The PcVg1 cDNA is 5748 nucleotides (nt) with a ...

  2. Citrus breeding, genetics and genomics in Japan.

    Science.gov (United States)

    Omura, Mitsuo; Shimada, Takehiko

    2016-01-01

    Citrus is one of the most cultivated fruits in the world, and satsuma mandarin (Citrus unshiu Marc.) is a major cultivated citrus in Japan. Many excellent cultivars derived from satsuma mandarin have been released through the improvement of mandarins using a conventional breeding method. The citrus breeding program is a lengthy process owing to the long juvenility, and it is predicted that marker-assisted selection (MAS) will overcome the obstacle and improve the efficiency of conventional breeding methods. To promote citrus molecular breeding in Japan, a genetic mapping was initiated in 1987, and the experimental tools and resources necessary for citrus functional genomics have been developed in relation to the physiological analysis of satsuma mandarin. In this paper, we review the progress of citrus breeding and genome researches in Japan and report the studies on genetic mapping, expression sequence tag cataloguing, and molecular characterization of breeding characteristics, mainly in terms of the metabolism of bio-functional substances as well as factors relating to, for example, fruit quality, disease resistance, polyembryony, and flowering. PMID:27069387

  3. Molecular detection of Candidatus liberibacter asiaticus associated with Citrus greening (Huanglongbing of mandarin by DNA template preparation

    Directory of Open Access Journals (Sweden)

    G.P. Jagtap

    2013-02-01

    Full Text Available The polymerase Chain Reaction (PCR diagnosis is more reliable and sensitive diagnostic tool for detecting greening bacterium than other conventional approaches like Electron microscopy, DNA-DNA hybridization and immunofluorescence (IF for detection of citrus greening. During experiments, it was observed that sodium sulphite method of DNA isolation provided higher yield and better quality DNA than other methods. Primer C (450 bp was more efficient in amplifying the DNA of greening bacterium even at a very low concentration of 0.1 pg. To confirm the reliability of PCR, the greening bacterium was also detected in graft-inoculated plants, which showed typical greening bacterium was also detected in graft-inoculated plants, which showed typical greening symptoms. Results showed amplification of 450 bp in PCR suggesting sampling in March is more suitable for PCR detection of greening bacterium.

  4. Progress on Dissecting and Controlling the Citrus Huanglongbing Complex

    OpenAIRE

    Duan, Yongping; Zhou, Lijuan; Zhang, Muqing; Benyon, Lesley; Armstrong-Vahling, Cheryl; Hoffman, Michele; Hao, Guixia; Zou, Huasong; Doud, Melissa; Ding, Fang; Morgan, Kent

    2014-01-01

    Citrus huanglongbing (HLB) is a century-old and emerging disease that impedes citrus production worldwide. ‘Candidatus Liberibacter asiaticus’ (Las) is the globally prevalent species of HLB bacteria. Here we describe our molecular characterizations of Las, and our newly-developed control methods for citrus HLB. From a genomics standpoint, we revealed Las has a significantly reduced genome (1.26Mb) and unique features adapted to its intracellular life style.  Although the genome is small, Las ...

  5. Molecular Characterization and Genetic Diversity Analysis of Sweet Orange Citrus sinensis L. Osbeck Cultivars in Iraq Using RAPD Markers

    Directory of Open Access Journals (Sweden)

    Ali Saeed Atiyah AL-Janabi

    2016-03-01

    Full Text Available Sweet orange (Citrus sinensis L. Osbeck is one of the most important commercially cultivated fruit crops of Citrus. Genetic diversity and inter-relationship among 5 cultivars (Indian, Iraqi, Japanese, Syrian, Egyptian of C. sinensis were analyzed based on RAPD markers. Six primers generated reproducible and easily storable RAPD profiles with a number of amplified DNA fragments ranging from 6 to 14 fragment bands. The total number of amplicons detected was 51, including 14 fragments unique bands with average reached 2.8 fragments ̸ primers. While the number of polymorphic ranged from 0 to 8 with an average reached 4.4 fragments ̸ primers with the polymorphic percentage ranged from 0% to 57.1%. While the number of monomorphic ranged from 2 to 5 fragment bands and was total of the monomorphic 15 fragments with an average reached 3 fragments ̸ primers with the monomorphic percentage was 14.2 % to 83.3%. A maximum numbers of amplicons was amplified with primer OPS-238 reached 14 fragments while the minimum number of fragments was amplified with primer OPS-253 reached 6 fragments. The highest number of polymorphic bands reached 8 fragments was obtained with primer OPS-238 with high percentage 57.1%, while the highest number of monomorphic bands reached 5 fragments with high percentage 83.3% was obtained with primer OPS-253. RAPD markers detected genetic distance and similarity, amaximum genetic distance value was observed between Japanese (Jap and Syrian (Syr cultivars reached 0.530 with less similarity value reached 47%, a minimum genetic distance value was observed between sweet Iraqi (Irq and Indian (Ind cultivars reached 0.239 with high similarity value reached 76.1%. The similarity matrices were employed in the cluster analysis to generate a dendrogram using the UPGMA method. The cluster tree analysis showed that the sweet orange cultivars were broadly divided into two main groups A and B with similarity reached 50%. A group including

  6. Effect of temperature on growth and HSP70 mRNA expres-sion of thermo-tolerant brood of Apostichopus japonicus%温度选择对刺参群体在不同温度下生长及热休克蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    刘广斌; 杨红生; 刘石林

    2010-01-01

    高温(26℃)培育刺参(Apostichopus japonicus)幼体获得选择群体, 生长至1.03 g±1.30 g, 与对照组幼参在18、20、22和24℃温度下培育.二者的成活率随着温度升高而降低, 而选择组幼参24℃的成活率显著高于对照群体(P<0.05).不同温度下的特定生长率之间没有差异.TRIzol法提取刺参总RNA,获得刺参HSP70片断基因序列, 利用实时定量PCR方法检测,选择群体HSP70mRNA的表达量在正常温度和热激条件下都较对照组高, 但表达量升高的倍数相近, 选择组为1.99倍, 对照组为2.07倍.

  7. Multiple molecular penumbras after focal cerebral ischemia.

    Science.gov (United States)

    Sharp, F R; Lu, A; Tang, Y; Millhorn, D E

    2000-07-01

    Though the ischemic penumbra has been classically described on the basis of blood flow and physiologic parameters, a variety of ischemic penumbras can be described in molecular terms. Apoptosis-related genes induced after focal ischemia may contribute to cell death in the core and the selective cell death adjacent to an infarct. The HSP70 heat shock protein is induced in glia at the edges of an infarct and in neurons often at some distance from the infarct. HSP70 proteins are induced in cells in response to denatured proteins that occur as a result of temporary energy failure. Hypoxia-inducible factor (HIF) is also induced after focal ischemia in regions that can extend beyond the HSP70 induction. The region of HIF induction is proposed to represent the areas of decreased cerebral blood flow and decreased oxygen delivery. Immediate early genes are induced in cortex, hippocampus, thalamus, and other brain regions. These distant changes in gene expression occur because of ischemia-induced spreading depression or depolarization and could contribute to plastic changes in brain after stroke. PMID:10908035

  8. Tracking the interplay between bound peptide and the lid domain of DnaK, using molecular dynamics.

    Science.gov (United States)

    Azoulay, Itzhaq; Kucherenko, Nataly; Nachliel, Esther; Gutman, Menachem; Azem, Abdussalam; Tsfadia, Yossi

    2013-01-01

    Hsp70 chaperones consist of two functional domains: the 44 kDa Nucleotide Binding Domain (NBD), that binds and hydrolyses ATP, and the 26 kDa Substrate Binding Domain (SBD), which binds unfolded proteins and reactivates them, utilizing energy obtained from nucleotide hydrolysis. The structure of the SBD of the bacterial Hsp70, DnaK, consists of two sub-domains: A β-sandwich part containing the hydrophobic cavity to which the hepta-peptide NRLLLTG (NR) is bound, and a segment made of 5 α-helices, called the "lid" that caps the top of the β-sandwich domain. In the present study we used the Escherichia coli Hsp70, DnaK, as a model for Hsp70 proteins, focusing on its SBD domain, examining the changes in the lid conformation. We deliberately decoupled the NBD from the SBD, limiting the study to the structure of the SBD section, with an emphasis on the interaction between the charges of the peptide with the residues located in the lid. Molecular dynamics simulations of the complex revealed significant mobility within the lid structure; as the structure was released from the forces operating during the crystallization process, the two terminal helices established a contact with the positive charge at the tip of the peptide. This contact is manifested only in the presence of electrostatic attraction. The observed internal motions within the lid provide a molecular role for the function of this sub-domain during the reaction cycle of Hsp 70 chaperones. PMID:23774839

  9. Tracking the Interplay between Bound Peptide and the Lid Domain of DnaK, Using Molecular Dynamics

    Directory of Open Access Journals (Sweden)

    Yossi Tsfadia

    2013-06-01

    Full Text Available Hsp70 chaperones consist of two functional domains: the 44 kDa Nucleotide Binding Domain (NBD, that binds and hydrolyses ATP, and the 26 kDa Substrate Binding Domain (SBD, which binds unfolded proteins and reactivates them, utilizing energy obtained from nucleotide hydrolysis. The structure of the SBD of the bacterial Hsp70, DnaK, consists of two sub-domains: A β-sandwich part containing the hydrophobic cavity to which the hepta-peptide NRLLLTG (NR is bound, and a segment made of 5 α-helices, called the “lid” that caps the top of the β-sandwich domain. In the present study we used the Escherichia coli Hsp70, DnaK, as a model for Hsp70 proteins, focusing on its SBD domain, examining the changes in the lid conformation. We deliberately decoupled the NBD from the SBD, limiting the study to the structure of the SBD section, with an emphasis on the interaction between the charges of the peptide with the residues located in the lid. Molecular dynamics simulations of the complex revealed significant mobility within the lid structure; as the structure was released from the forces operating during the crystallization process, the two terminal helices established a contact with the positive charge at the tip of the peptide. This contact is manifested only in the presence of electrostatic attraction. The observed internal motions within the lid provide a molecular role for the function of this sub-domain during the reaction cycle of Hsp 70 chaperones.

  10. 跑台运动训练对脑缺血损伤大鼠热休克蛋白70及C-MYC表达的影响%Effects of treadmill training on the expression of HSP70 and C-MYC in the brains of rats with focal cerebral ischemia

    Institute of Scientific and Technical Information of China (English)

    刘德山; 刘楠; 张逸仙; 杜厚伟; 陈荣华

    2010-01-01

    目的 探讨跑台运动训练对局灶性脑缺血大鼠神经功能恢复和脑缺血组织中热休克蛋白70(HSP70)及C-MYC表达的影响.方法 将42只清洁级成年雄性SD大鼠分为假手术组6只、模型组18只、运动组18只.模型组、运动组大鼠采用改良的Longs线栓法制备大脑中动脉闭塞(MCAO)脑缺血模型,运动组于造模成功后24 h采用跑台训练器进行运动训练,每周6 d,共2周,其余2组则置于普通笼内饲养,期间可自由活动、进食.采用修正的神经行为学评分方法评价模型组和运动组大鼠MCAO脑缺血后第3,7,14天的神经功能,并断头取脑,采用逆转录-多聚酶链反应(RT-PCR)法、免疫组织化学和Western blot法检测脑缺血组织中HSP70、C-MYC的表达.结果 运动组大鼠脑缺血第7,14天神经功能评分均明显优于模型组(P<0.05或0.01);运动组大鼠脑缺血第7,14天HSF70和C-MYC表达较模型组增强(P<0.05或0.01).结论 运动训练可促进脑缺血大鼠神经功能恢复,其机制可能与上调脑缺血组织中HSP70及C-MYC表达有关.%Objective To observe the effects of treadmill training on the recovery of neurological function and the expression of HSP70 and C-MYC in the brains of rats with focal cerebral ischemia. Methods Forty-two male adult Sprague-Dawley rats were randomly divided into a sham group ( n =6), a model group (n =18) and a treadmill exercise group (n=18). Focal cerebral ischemia was induced by right middle cerebral artery occlusion (MCAO) in the model group and exercise group using a modified version of Longa's method. The rats in the treadmill exercise group were given treadmill training 6 d per week for 2 weeks after 24 h of MCAO. By contrast, the rats in the sham group and the model group were reared in standard cages. Before the rats were sacrificed at the 3rd, 7th and 14th d after MCAO, their neurological functions were tested using modified neurological severity scores ( mNSS) , and the mRNA and

  11. Physicochemical, mechanical, and molecular properties of nonlysogenic and p22-lysogenic Salmonella typhimurium treated with citrus oil.

    Science.gov (United States)

    Ahn, Juhee; Almario, Jose Alejandro; Salaheen, Serajus; Biswas, Debabrata

    2014-05-01

    The aim of this study was to evaluate the phenotypic and genotypic properties of nonlysogenic Salmonella Typhimurium (ST(P22-)) and lysogenic Salmonella Typhimurium (ST(P22+)) in the presence of sublethal concentrations (SLC2D) of citrus essential oils (CEOs), which were used to evaluate antimicrobial susceptibility, cell surface hydrophobicity, autoaggregation ability, bacterial motility, lysogenic conversion, gene expression patterns, and antibiofilm formation. The SLC2D values of non-heat-treated (N-CEO) and heat-treated (H-CEO) CEO in an autoclave at 121°C for 20 min were 2.0 to 2.1 mg/ml against ST(P22-) and 1.7 to 1.9 mg/ml against STP(22+). The rates of injured ST(P22-) and ST(P22+) cells treated with SLC2D of N-CEO and H-CEO ranged from 67 to 83%. The hydrophobicity and autoaggregation were decreased to 2.5 and 19.5% for ST(P22-) and 4.7 and 21.7% for ST(P22+), respectively, in the presence of N-CEO. A noticeable reduction in the swarming motility was observed in ST(P22-) with N-CEO (14.5%) and H-CEO (13.3%). The numbers of CEO-induced P22 were 5.40 log PFU/ml for N-CEO and 5.65 log PFU/ml for H-CEO. The relative expression of hilA, hilC, hilD, invA, invC, invE, invF, sirA, and sirB was down-regulated in ST(P22-) and ST(P22+) with N-CEO and H-CEO. The numbers of adherent ST(P22-) and ST(P22+) were effectively reduced by more than 1 log in the presence of CEO. These results suggest that CEO has potential to be used to control bacterial attachment, colonization, and invasion. PMID:24780330

  12. Molecular characterization of SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL gene family from Citrus and the effect of fruit load on their expression

    Directory of Open Access Journals (Sweden)

    Liron eShalom

    2015-05-01

    Full Text Available We recently identified a Citrus gene encoding SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL transcription factor that contained a sequence complementary to miR156. Genes of the SPL family are known to play a role in flowering regulation and phase transition. In Citrus, the mRNA levels of the gene were significantly altered by fruit load in buds; under heavy fruit load (ON-Crop trees, known to suppress next year flowering, the mRNA levels were down-regulated, while fruit removal (de-fruiting, inducing next-year flowering, resulted in its up-regulation. In the current work, we set on to study the function of the gene. We showed that the Citrus SPL was able promote flowering independently of photoperiod in Arabidopsis, while miR156 repressed its flowering-promoting activity. In order to find out if fruit load affected the expression of additional genes of the SPL family, we identified and classified all SPL members in the Citrus genome, and studied their seasonal expression patterns in buds and leaves, and in response to de-fruiting. Results showed that two additional SPL-like genes and miR172, known to be induced by SPLs in Arabidopsis, were altered by fruit load. The relationships between these factors in relation to the fruit-load effect on Citrus flowering are discussed.

  13. Molecular characterization of SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) gene family from Citrus and the effect of fruit load on their expression.

    Science.gov (United States)

    Shalom, Liron; Shlizerman, Lyudmila; Zur, Naftali; Doron-Faigenboim, Adi; Blumwald, Eduardo; Sadka, Avi

    2015-01-01

    We recently identified a Citrus gene encoding SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factor that contained a sequence complementary to miR156. Genes of the SPL family are known to play a role in flowering regulation and phase transition. In Citrus, the mRNA levels of the gene were significantly altered by fruit load in buds; under heavy fruit load (ON-Crop trees), known to suppress next year flowering, the mRNA levels were down-regulated, while fruit removal (de-fruiting), inducing next-year flowering, resulted in its up-regulation. In the current work, we set on to study the function of the gene. We showed that the Citrus SPL was able promote flowering independently of photoperiod in Arabidopsis, while miR156 repressed its flowering-promoting activity. In order to find out if fruit load affected the expression of additional genes of the SPL family, we identified and classified all SPL members in the Citrus genome, and studied their seasonal expression patterns in buds and leaves, and in response to de-fruiting. Results showed that two additional SPL-like genes and miR172, known to be induced by SPLs in Arabidopsis, were altered by fruit load. The relationships between these factors in relation to the fruit-load effect on Citrus flowering are discussed. PMID:26074947

  14. Citrus Waste Biomass Program

    Energy Technology Data Exchange (ETDEWEB)

    Karel Grohman; Scott Stevenson

    2007-01-30

    Renewable Spirits is developing an innovative pilot plant bio-refinery to establish the commercial viability of ehtanol production utilizing a processing waste from citrus juice production. A novel process based on enzymatic hydrolysis of citrus processing waste and fermentation of resulting sugars to ethanol by yeasts was successfully developed in collaboration with a CRADA partner, USDA/ARS Citrus and Subtropical Products Laboratory. The process was also successfully scaled up from laboratory scale to 10,000 gal fermentor level.

  15. Molecular characterization of SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) gene family from Citrus and the effect of fruit load on their expression

    OpenAIRE

    Shalom, Liron; Shlizerman, Lyudmila; Zur, Naftali; Doron-Faigenboim, Adi; Blumwald, Eduardo; Sadka, Avi

    2015-01-01

    We recently identified a Citrus gene encoding SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factor that contained a sequence complementary to miR156. Genes of the SPL family are known to play a role in flowering regulation and phase transition. In Citrus, the mRNA levels of the gene were significantly altered by fruit load in buds; under heavy fruit load (ON-Crop trees), known to suppress next year flowering, the mRNA levels were down-regulated, while fruit removal (de-fruiting),...

  16. Founder lines for improved citrus biotechnology

    Science.gov (United States)

    This article discusses the research needed to develop the RMCE strategy and molecular assays for site-specific recombinases as tools for genome manipulation. Explanation of genetic engineering used to generate transgenic citrus plants to exhibit a novel phenotype, but not to contain the recombinase...

  17. α-Mangostin from Cratoxylum arborescens demonstrates apoptogenesis in MCF-7 with regulation of NF-κB and Hsp70 protein modulation in vitro, and tumor reduction in vivo

    OpenAIRE

    Ibrahim MY; Hashim NM; Mohan S; Abdulla MA; Kamalidehghan B; Ghaderian M; Dehghan F; Ali LZ; Arbab IA; Yahayu M; Lian GE; Ahmadipour F; Ali HM

    2014-01-01

    Mohamed Yousif Ibrahim,1 Najihah Mohd Hashim,1 Syam Mohan,2 Mahmood Ameen Abdulla,3 Behnam Kamalidehghan,1 Mostafa Ghaderian,1,4 Firouzeh Dehghan,1,5 Landa Zeenelabdin Ali,1 Ismail Adam Arbab,6 Maizatulakmal Yahayu,7 Gwendoline Ee Cheng Lian,8 Fatemeh Ahmadipour,1 Hapipah Mohd Ali9 1Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia; 2Medical Research Centre, Jazan University, Jazan, Saudi Arabia; 3Department of Molecular Medicine, Faculty of Medicine Un...

  18. α-Mangostin from Cratoxylum arborescens demonstrates apoptogenesis in MCF-7 with regulation of NF-κB and Hsp70 protein modulation in vitro, and tumor reduction in vivo

    OpenAIRE

    Ibrahim, Mohamed Yousif

    2014-01-01

    Mohamed Yousif Ibrahim,1 Najihah Mohd Hashim,1 Syam Mohan,2 Mahmood Ameen Abdulla,3 Behnam Kamalidehghan,1 Mostafa Ghaderian,1,4 Firouzeh Dehghan,1,5 Landa Zeenelabdin Ali,1 Ismail Adam Arbab,6 Maizatulakmal Yahayu,7 Gwendoline Ee Cheng Lian,8 Fatemeh Ahmadipour,1 Hapipah Mohd Ali9 1Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia; 2Medical Research Centre, Jazan University, Jazan, Saudi Arabia; 3Department of Molecular Medicine, Faculty of Medicine Un...

  19. Involvement of NF-ΚB and HSP70 signaling pathways in the apoptosis of MDA-MB-231 cells induced by a prenylated xanthone compound, α-mangostin, from Cratoxylum arborescens [Corrigendum

    OpenAIRE

    Ibrahim, Mohamed Yousif

    2015-01-01

    Ibrahim MY, Hashim NM, Mohan S, et al. Drug Des Devel Ther. 2014;8:2193–2211. On page 2193, author affiliations, “Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia” should be “Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia”. Introduction, first paragraph, the text should read: “Breast cancer has become a major cause of morb...

  20. Involvement of NF-ΚB and HSP70 signaling pathways in the apoptosis of MDA-MB-231 cells induced by a prenylated xanthone compound, α-mangostin, from Cratoxylum arborescens

    OpenAIRE

    Ibrahim, Mohamed Yousif

    2014-01-01

    Mohamed Yousif Ibrahim,1 Najihah Mohd Hashim,1 Syam Mohan,2 Mahmood Ameen Abdulla,3 Siddig Ibrahim Abdelwahab,2 Behnam Kamalidehghan,1 Mostafa Ghaderian,1,4 Firouzeh Dehghan,1,5 Landa Zeenelabdin Ali,1 Hamed Karimian,1 Maizatulakmal Yahayu,6 Gwendoline Cheng Lian Ee,7 Abdoreza Soleimani Farjam,8 Hapipah Mohd Ali9 1Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia; 2Medical Research Centre, Jazan University, Jazan, Saudi Arabia; 3Department of Molecular ...

  1. Consumption of vitamin B6 reduces colonic damage and protein expression of HSP70 and HO-1, the anti-tumor targets, in rats exposed to 1,2-dimethylhydrazine

    OpenAIRE

    KAYASHIMA, TOMOKO; Tanaka, Kenta; OKAZAKI, YUKAKO; Matsubara, Kiminori; Yanaka, Noriyuki; Kato, Norihisa

    2011-01-01

    Mounting evidence indicates that vitamin B6 is a protective factor for colon cancer. Elevations in colonic damage, cell proliferation and heat shock proteins (HSPs, molecular chaperones) have been suggested to be associated with colon carcinogenesis. This study was performed to examine the effect of dietary levels of vitamin B6 (1, 7 or 35 mg pyridoxine HCl/kg diet) for 22 weeks on colon damage, epithelial cell proliferation and expression of HSPs in rats exposed to 1,2-dimethylhydrazine (DMH...

  2. A comparative analysis of different molecular targets using PCR for diagnosis of old world leishmaniasis.

    Science.gov (United States)

    Koltas, Ismail S; Eroglu, Fadime; Uzun, Soner; Alabaz, Derya

    2016-05-01

    The different sensitivity values were obtained in each study conducted for the diagnosis of leishmaniasis with the polymerase chain reaction (PCR). However, a standardized PCR target for the diagnosis of leishmaniasis does not exist. The aim of the current study, the most ideal PCR target was determined for diagnosis of leishmaniasis. A total of 72 smear and 48 bone marrow samples were analyzed with six different molecular targets to determine their potential as a tool for the specific molecular diagnosis of leishmaniasis using PCR. The positivity-negativity value and the sensitivity-specificity of each PCR targets were calculated. The positivity value of PCR targets were sequenced in different levels in the diagnosis of leishmaniasis from highest to lowest in the order of kDNA-PCR > SSU rRNA-PCR > ITS2-PCR > ITS1-PCR > ME-PCR > HSP70-PCR. The sensitivities of PCR targets except ITS1-PCR, ME-PCR and HSP70-PCR were found to be 100% in cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) cases as compared to microscopic examination accepted as a gold standard. The sensitivities of ITS1-PCR, ME-PCR and HSP70-PCR were found 96.6%, 90.0% and 86.6%, respectively, in CL-cases. In addition, the sensitivities of ITS1-PCR, ME-PCR and HSP70-PCR were found 90.0%, 70.0% and 60.0%, respectively, in VL-cases. The kDNA genomic region was the most sensitive for routine diagnosis of leishmaniasis. ITS1-PCR restriction fragment length polymorphism, the alternative method for the identification of Old World Leishmania species, did not require culturing of the parasites. PMID:26896641

  3. Analyses of proteomic expression profiles and nutrient status of citrus plants in response to Huanglongbing

    OpenAIRE

    H. Lin; Nwugo, C. C.; Duan, Y.

    2014-01-01

    Huanglongbing (HLB) is a highly devastating citrus disease and represents a major threat to the citrus industries in US. The etiology of HLB worldwide is associated with three insect-transmissible phloem-limited members of the bacterial group ‘Candidatus Liberibacter spp.’, prevalently ‘Candidatus Liberibacter asiaticus’ (Las). All citrus cultivars are susceptible to HLB. To better understand the physiological and molecular processes involved in host responses to Las, proteomic analyses via 2...

  4. Citrus pectin as a hydrocolloid emulsifier: Emulsifying and emulsion stabilizing properties

    OpenAIRE

    Schmidt, Ulrike

    2016-01-01

    Citrus pectin is commonly used as a gelling agent and viscosity enhancer in food products. However, it is considered to be less effective in terms of emulsifier. This thesis explores ways to enhance the emulsifying properties of citrus pectin by molecular modification and by a proper choice of recipe parameters. It is shown how stable emulsions can be produced with citrus pectin when the viscosity enhancing effects are properly accounted for.

  5. Involvement of NF-κB and HSP70 signaling pathways in the apoptosis of MDA-MB-231 cells induced by a prenylated xanthone compound, α-mangostin, from Cratoxylum arborescens [Corrigendum

    Directory of Open Access Journals (Sweden)

    Ibrahim MY

    2015-06-01

    Full Text Available Ibrahim MY, Hashim NM, Mohan S, et al. Drug Des Devel Ther. 2014;8:2193–2211. On page 2193, author affiliations, “Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia” should be “Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia”. Introduction, first paragraph, the text should read: “Breast cancer has become a major cause of morbidity and mortality in women globally. The American Cancer Society (ACS reported that breast cancer incidence has an estimation of 26% of all new cancer cases, which is the highest in ratio among all the cancers in American women.1 The National Cancer Registry (NCR in Malaysia has reported that one in twenty Malaysian women are at a risk of acquiring breast in their lifetime.2 The incidence rate in Malaysia is still considered low if compared to Europe and United States.3 Up to 70% of breast cancer development causes occur in women is reported to be of environmental factors and lifestyle.4,5”  Introduction, second paragraph, first sentence, the text should read: “Radiation therapy has become a valuable tool among cancer treatment strategies for the control of local and regional diseases after 1960 with the invention of the linear accelerator, but, like surgery, radiation therapy alone cannot enucleate metastatic cancer.”  Discussion, first paragraph, first sentence, the text should read: “Apoptosis has a vital role in many functions, ranging from fetal development to adult tissue homeostasis.32”   Read the original article

  6. Phylogenetic Analysis of Citrus tristeza virus Isolates of Wild Type Citrus in China

    Institute of Scientific and Technical Information of China (English)

    YI Long; ZHOU Chang-yong

    2014-01-01

    The genetic variation and phylogenetic relationships of Citrus tristeza virus (CTV) isolates collected from Chinese wild type citrus were analyzed by comparing the sequences of nine genomic regions (p23, p20, p13, p18, p25, p27, POL, HEL and k17) with the CTV isolates of cultivated citrus from different countries. The results showed that the divergence pattern of genomic RNA of the CTV isolates from wild type citrus was similar to that of other isolates from cultivated citrus, the 3´ proximal region was relatively conserved, and the 5´ proximal region had greater variability. The nine genomic regions of CTV isolates analyzed were found to have been under purifying selection in the evolution process. Phylogenetic analysis showed that the eleven Chinese wild CTV isolates were located at different clades and did not relfect their geographical origins, suggesting genetic diversity among the Chinese wild CTV populations. These results will aid in the understanding of molecular evolution of the Chinese CTV populations.

  7. PHARMACOLOGICAL IMPORTANCE OF CITRUS FRUITS

    Directory of Open Access Journals (Sweden)

    Amita Tomar *, Mridula Mall and Pragya Rai

    2013-01-01

    Full Text Available This paper reviews the pharmacological importance of citrus fruits. Citrus fruits are used for various pharmacological importance. According to literature the citrus fruit possess anti-cancer, antimicrobial, antioxidant, antiulcer, anti-inflammatory, and hypolipidemic and hepatoprotective properties.

  8. Molecular and physiological changes in response to salt stress in Citrus macrophylla W plants overexpressing Arabidopsis CBF3/DREB1A.

    Science.gov (United States)

    Alvarez-Gerding, Ximena; Espinoza, Carmen; Inostroza-Blancheteau, Claudio; Arce-Johnson, Patricio

    2015-07-01

    Plant stress induced by high salinity has leading to an important reduction in crop yields. Due to their tropical origin, citrus fruits are highly sensitive to salts. Rootstocks are the root system of fruit trees, regulating ion uptake and transport to the canopy. Therefore, increasing their salt tolerance could improve the salt tolerance of the fruit tree. For this, we genetically-transformed an important rootstock for lemon, Citrus macrophylla W, to constitutively express the CBF3/DREB1A gene from Arabidopsis, a well-studied salinity tolerance transcription factor. Transgenic lines showed normal size, with no dwarfism. Under salt stress, some transgenic lines showed greater growth, similar accumulation of chloride and sodium in the leaves and better stomatal conductance, in comparison to wild-type plants. Quantitative real-time analyses showed a similar expression of several CBF3/DREB1A target genes, such as COR15A, LEA 4/5, INV, SIP1, P5CS, GOLS, ADC2 and LKR/SDH, in transgenic lines and wild type plants, with the exception of INV that shows increased expression in line 4C15. Under salt stress, all measured transcript increased in both wild type and transgenics lines, with the exception of INV. Altogether, these results suggest a higher salt tolerance of transgenic C. macrophylla plants induced by the overexpression of AtCBF3/DREB1A. PMID:25914135

  9. 78 FR 63369 - Citrus Canker, Citrus Greening, and Asian Citrus Psyllid; Interstate Movement of Regulated...

    Science.gov (United States)

    2013-10-24

    ... Animal and Plant Health Inspection Service 7 CFR Part 301 RIN 0579-AD29 Citrus Canker, Citrus Greening... published an interim rule \\1\\ in the Federal Register (76 FR 23449-23459, Docket No. APHIS-2010-0048) that....usda.gov/plant_health/plant_pest_info/citrus/index.shtml and on Regulations.gov (see footnote 1...

  10. 75 FR 17289 - Citrus Seed Imports; Citrus Greening and Citrus Variegated Chlorosis

    Science.gov (United States)

    2010-04-06

    ... Animal and Plant Health Inspection Service 7 CFR Part 319 RIN 0579-AD07 Citrus Seed Imports; Citrus... of nursery stock to prohibit the importation of propagative seed of several Rutaceae (citrus family... are also requiring propagative seed of these genera imported from all other countries to...

  11. Antioxidant activity of Citrus fruits.

    Science.gov (United States)

    Zou, Zhuo; Xi, Wanpeng; Hu, Yan; Nie, Chao; Zhou, Zhiqin

    2016-04-01

    Citrus is well-known for its nutrition and health-promotion values. This reputation is derived from the studies on the biological functions of phytochemicals in Citrus fruits and their derived products in the past decades. In recent years, the antioxidant activity of Citrus fruits and their roles in the prevention and treatment of various human chronic and degenerative diseases have attracted more and more attention. Citrus fruits are suggested to be a good source of dietary antioxidants. To have a better understanding of the mechanism underlying the antioxidant activity of Citrus fruits, we reviewed a study on the antioxidant activity of the phytochemicals in Citrus fruits, introduced methods for antioxidant activity evaluation, discussed the factors which influence the antioxidant activity of Citrus fruits, and summarized the underlying mechanism of action. Some suggestions for future study were also presented. PMID:26593569

  12. Activation of phospholipase A2 by Hsp70 in vitro

    DEFF Research Database (Denmark)

    Mahalka, Ajay K; Code, Christian; Rezaijahromi, Behnam; Kirkegaard, Thomas; Jaattela, Marja; Kinnunen, Paavo

    2011-01-01

    We recently suggested a novel mechanism for the activation of phospholipase A2 (PLA2), with a (catalytically) highly active oligomeric state, which subsequently becomes inactivated by conversion into amyloid. This process can be activated by lysophosphatidylcholine which promotes both...

  13. Glyoxalase I drives epithelial-to-mesenchymal transition via argpyrimidine-modified Hsp70, miR-21 and SMAD signalling in human bronchial cells BEAS-2B chronically exposed to crystalline silica Min-U-Sil 5: Transformation into a neoplastic-like phenotype.

    Science.gov (United States)

    Antognelli, Cinzia; Gambelunghe, Angela; Muzi, Giacomo; Talesa, Vincenzo Nicola

    2016-03-01

    Glyoxalase I (Glo1) is the main scavenging enzyme of methylglyoxal (MG), a potent precursor of advanced glycation end products (AGEs). AGEs are known to control multiple biological processes, including epithelial to mesenchymal transition (EMT), a multistep phenomenon associated with cell transformation, playing a major role in a variety of diseases, including cancer. Crystalline silica is a well-known occupational health hazard, responsible for a great number of human pulmonary diseases, such as silicosis. There is still much debate concerning the carcinogenic role of crystalline silica, mainly due to the lack of a causal demonstration between silica exposure and carcinogenesis. It has been suggested that EMT might play a role in crystalline silica-induced lung neoplastic transformation. The aim of this study was to investigate whether, and by means of which mechanism, the antiglycation defence Glo1 is involved in Min-U-Sil 5 (MS5) crystalline silica-induced EMT in BEAS-2B human bronchial epithelial cells chronically exposed, and whether this is associated with the beginning of a neoplastic-like transformation process. By using gene silencing/overexpression and scavenging/inhibitory agents, we demonstrated that MS5 induced hydrogen peroxide-mediated c-Jun-dependent Glo1 up-regulation which resulted in a decrease in the Argpyrimidine-modified Hsp70 protein level which triggered EMT in a novel mechanism involving miR-21 and SMAD signalling. The observed EMT was associated with a neoplastic-like phenotype. The results obtained provide a causal in vitro demonstration of the MS5 pro-carcinogenic transforming role and more importantly they provide new insights into the mechanisms involved in this process, thus opening new paths in research concerning the in vivo study of the carcinogenic potential of crystalline silica. PMID:26784015

  14. Sequence analysis of three citrus viroids infecting a single Tunisian citrus tree (Citrus, reticulata, Clementine

    Directory of Open Access Journals (Sweden)

    Amine Elleuch

    2006-01-01

    Full Text Available We report the nucleotide sequences of three citrus viroids belonging to three different genera: Citrus exocortis viroid (CEVd, Hop stunt viroid (HSVd and Citrus viroid-III (CVd-III isolated from a single natural infected Citrus reticulata var. Clementine tree growing in a tree nursery in Manouba (near Tunis Capital. We describe the sequence variability of these viroids from their natural host without using an alternative passage by an indicator host or an artificial inoculation. This work confirms that naturally occurring viroid infections contain a mixture of sequence variants. These are the first sequences of citrus viroids from Africa.

  15. Citrus stubborn disease (CSD)

    Science.gov (United States)

    CSD is caused by Spiroplasma citri, a phloem-limited, cell-wall-less bacterium. S. citri is transmitted in a propagative, circulative manner by several leafhoppers including Circulifer tenellus and Scaphytopius nitridus in citrus-growing regions of California and Arizona and by C. haematoceps (syn....

  16. Potential Nutritional Benefits of Current Citrus Consumption

    OpenAIRE

    Tami Turner; Burri, Betty J.

    2013-01-01

    Citrus contains nutrients and phytochemicals that may be beneficial for health. We collected citrus production and consumption data and estimated the amount of these compounds that are consumed. We then compared the amounts of citrus and citrus-derived compounds used in studies that suggest a health benefit to the amounts typically found in citrus. Data is scarce, but suggests that citrus consumption might improve indices of antioxidant status, and possibly cardiovascular health and insulin s...

  17. Potential Nutritional Benefits of Current Citrus Consumption

    Directory of Open Access Journals (Sweden)

    Tami Turner

    2013-03-01

    Full Text Available Citrus contains nutrients and phytochemicals that may be beneficial for health. We collected citrus production and consumption data and estimated the amount of these compounds that are consumed. We then compared the amounts of citrus and citrus-derived compounds used in studies that suggest a health benefit to the amounts typically found in citrus. Data is scarce, but suggests that citrus consumption might improve indices of antioxidant status, and possibly cardiovascular health and insulin sensitivity.

  18. 分子伴侣对酵母朊病毒[ PSI+]增殖影响的研究进展%Advances in molecular chaperones regulating yeast prion [ PSI+] propagation

    Institute of Scientific and Technical Information of China (English)

    林康伟; 连惠勇; 蔡澎

    2015-01-01

    The finding and research on yeast prion are of great values for biology and medical sciences.Research advances in molecular chaperones, especially in Hsp104p, Hsp70p and Hsp40p, regulating yeast prion [PSI+] propaga-tion,are reviewed.%酵母朊病毒( prion)的发现和研究在生物学和医学上有着极其重要的理论价值和实际应用价值。该文综述了分子伴侣对酵母prion[PSI+]增殖影响的研究进展,重点介绍了热休克蛋白Hsp104p、Hsp70p和Hsp40p在其中发挥的重要作用。

  19. Taxonomy and phylogeny of the genus citrus based on the nuclear ribosomal dna its region sequence

    International Nuclear Information System (INIS)

    The genus Citrus (Aurantioideae, Rutaceae) is the sole source of the citrus fruits of commerce showing high economic values. In this study, the taxonomy and phylogeny of Citrus species is evaluated using sequence analysis of the ITS region of nrDNA. This study is based on 26 plants materials belonging to 22 Citrus species having wild, domesticated, and cultivated species. Through DNA alignment of the ITS sequence, ITS1 and ITS2 regions showed relatively high variations of sequence length and nucleotide among these Citrus species. According to previous six-tribe discrimination theory by Swingle and Reece, the grouping in our ITS phylogenetic tree reconstructed by ITS sequences was not related to tribe discrimination but species discrimination. However, the molecular analysis could provide more information on citrus taxonomy. Combined with ITS sequences of other subgenera in then true citrus fruit tree group, the ITS phylogenetic tree indicated subgenera Citrus was monophyletic and nearer to Fortunella, Poncirus, and Clymenia compared to Microcitrus and Eremocitrus. Abundant sequence variations of the ITS region shown in this study would help species identification and tribe differentiation of the genus Citrus. (author)

  20. Molecular Characterization and Expression Analysis of S1 self-incompatibility Locus-linked Pollen 3.15 Gene in Citrus reticulata

    Institute of Scientific and Technical Information of China (English)

    Hong Xia Miao; Zi Xing ye; Yong Hua Qin; Gui Bing Hu

    2013-01-01

    Gametophytic self-incompatibility (GSI) is controlled by a highly polymorphic locus called the S-locus,which is an important factor that can result in seedless fruit in Citrus.The S,self-incompatibility locuslinked pollen 3.15 gene (S,-3.15) belongs to a type of S locus gene.The role of S,-3.15 in the Sl reaction of Citrus has not yet been reported.In this study,full-length sequences of cDNA and DNA encoding the S,-3.15gene,referred to as CrS1-3.15,were isolated from ‘Wuzishatangju’ (Self-incompatibility,Sl) and ‘Shatangju’(Self-compatibility,SC).The predicted amino acid sequences of CrS1-3.15 between ‘Wuzishatangju’ and ‘Shatangju’ differ by only three amino acids.Compared to ‘Wuzishatangju’,three bases were substituted in the genomic DNA of CrS1-3.15 from ‘Shatangju’.Southern blot results showed that one copy of CrS1-3.15 existed in the genomic DNA of both ‘Wuzishatangju’ and ‘Shatangju’.The expression level of the CrS1-3.15 gene in the ovaries of ‘Shatangju’ was approximately 60-fold higher than that in the ovaries of ‘Wuzishatangju’.When ‘Wuzishatangju’ was cross-pollinated,the expression of CrS1-3.15 was upregulated in the ovaries at 3 d,and the highest expression levels were detected in the ovaries at 6 d after cross-pollination of ‘Wuzishatangju’ x ‘Shatangju’.To obtain the CrS,-3.15 protein,the full-length cDNA of CrS1-3.15 genes from ‘Wuzishatangju’ and ‘Shatangju’ was successfully expressed in Pichia pastoris.Pollen germination frequency of ‘Wuzishatangju’ was inhibited significantly with increasing CrS,-3.15 protein concentrations from Sl ‘Wuzishatangju’.

  1. A transcriptomic approach highlights induction of secondary metabolism in citrus fruit in response to Penicillium digitatum infection

    OpenAIRE

    González-Candelas Luis; Alamar Santiago; Sánchez-Torres Paloma; Zacarías Lorenzo; Marcos Jose F

    2010-01-01

    Abstract Background Postharvest losses of citrus fruit due to green mold decay, caused by the fungus Penicillium digitaum, have a considerable economic impact. However, little is known about the molecular processes underlying the response of citrus fruit to P. digitatum. Results Here we describe the construction of a subtracted cDNA library enriched in citrus genes preferentially expressed in response to pathogen infection followed by cDNA macroarray hybridization to investigate gene expressi...

  2. Impact of different temperatures on survival and energy metabolism in the Asian citrus psyllid, Diaphorina citri Kuwayama.

    Science.gov (United States)

    El-Shesheny, Ibrahim; Hijaz, Faraj; El-Hawary, Ibrahim; Mesbah, Ibrahim; Killiny, Nabil

    2016-02-01

    Temperature influences the life history and metabolic parameters of insects. Asian citrus psyllid (ACP), Diaphorina citri is a tropical and subtropical pest. ACP invaded new regions around the world and threatened the citrus industry as a vector for Huanglongbing (HLB) disease. ACP is widely distributed and can survive high (up to 45 °C) and low temperatures (as low as -6 °C). The precise mechanism of temperature tolerance in ACP is poorly understood. We investigated adult survival, cellular energy balance, gene expression, and nucleotide and sugar-nucleotide changes under the effect of different temperature regimes (0 °C to 45 °C with 5 °C intervals). The optimum temperatures for survival were 20 and 25 °C. Low temperatures of 0 °C and 5 °C caused 50% mortality after 2 and 4 days respectively, while one day at high temperature (40 °C and 45 °C) caused more than 95% mortality. The lowest quantity of ATP (3.69 ± 1.6 ng/insect) and the maximum ATPase enzyme activities (57.43 ± 7.6 μU/insect) were observed at 25 °C. Correlation between ATP quantities and ATPase activity was negative. Gene expression of hsp 70, V-type proton ATPase catalytic subunit A and ATP synthase α subunit matched these results. Twenty-four nucleotides and sugar-nucleotides were quantified using HPLC in ACP adults maintained at low, high, and optimum temperatures. The nucleotide profiles were different among treatments. The ratios between AMP:ATP and ADP:ATP were significantly decreased and positively correlated to adults survival, whereas the adenylate energy charge was increased in response to low and high temperatures. Exploring energy metabolic regulation in relation with adult survival might help in understanding the physiological basis of how ACP tolerates newly invaded regions. PMID:26603556

  3. Phytochemical analysis and radical scavenging profile of juices of Citrus sinensis, Citrus anrantifolia, and Citrus limonum

    OpenAIRE

    Rauf, Abdur; Uddin, Ghias; Ali, Jawad

    2014-01-01

    Background The aim of the current investigation was to identify bioactive secondary metabolites including phenols, tannins, flavonoids, terpinedes, and steroids and compare the phytochemical analysis and antioxidant profile of the juice extracted from the fruits of Citrus sinensis, Citrus anrantifolia, and Citrus limonum. Results Phytochemical screening is important for the isolation of new, novel, and rare secondary metabolites before bulk extraction. Phytochemical analysis of the desired pl...

  4. Valuing Catastrophic Citrus Losses

    OpenAIRE

    Adams, Damian C.; Kilmer, Richard L.; Moss, Charles B.; Schmitz, Andrew

    2004-01-01

    Courts are often required to estimate changes in welfare to agricultural operations from catastrophic events. For example, courts must assign damages in lawsuits, such as with pesticide drift cases, or determine "just compensation" when the government takes private land for public use, as with the removal of dairy farms from environmentally sensitive land or destruction of canker-contaminated citrus trees. In economics, the traditional method of quantifying producer losses is estimating chang...

  5. Inhibiting ethylene perception with 1-methylcyclopropene triggers molecular responses aimed to cope with cell toxicity and increased respiration in citrus fruits.

    Science.gov (United States)

    Establés-Ortiz, Beatriz; Romero, Paco; Ballester, Ana-Rosa; González-Candelas, Luis; Lafuente, María T

    2016-06-01

    The ethylene perception inhibitor 1-methylcyclopropene (1-MCP) has been critical in understanding the hormone's mode of action. However, 1-MCP may trigger other processes that could vary the interpretation of results related until now to ethylene, which we aim to understand by using transcriptomic analysis. Transcriptomic changes in ethylene and 1-MCP-treated 'Navelate' (Citrus sinensis L. Osbeck) oranges were studied in parallel with changes in ethylene production, respiration and peel damage. The effects of compounds modifying the levels of the ethylene co-product cyanide and nitric oxide (NO) on fruit physiology were also studied. Results suggested that: 1) The ethylene treatment caused sub-lethal stress since it induced stress-related responses and reduced peel damage; 2) 1-MCP induced ethylene-dependent and ethylene-independent responsive networks; 3) 1-MCP triggered ethylene overproduction, stress-related responses and metabolic shifts aimed to cope with cell toxicity, which mostly affected to the inner part of the peel (albedo); 4) 1-MCP increased respiration and drove metabolism reconfiguration for favoring energy conservation but up-regulated genes related to lipid and protein degradation and triggered the over-expression of genes associated with the plasma membrane cellular component; 5) Xenobiotics and/or reactive oxygen species (ROS) might act as signals for defense responses in the ethylene-treated fruit, while their uncontrolled generation would induce processes mimicking cell death and damage in 1-MCP-treated fruit; 6) ROS, the ethylene co-product cyanide and NO may converge in the toxic effects of 1-MCP. PMID:26990405

  6. Overexpression of a Modified Plant Thionin Enhances Disease Resistance to Citrus Canker and Huanglongbing (HLB).

    Science.gov (United States)

    Hao, Guixia; Stover, Ed; Gupta, Goutam

    2016-01-01

    Huanglongbing (HLB or citrus greening disease) caused by Candidatus Liberibacter asiaticus (Las) is a great threat to the US citrus industry. There are no proven strategies to eliminate HLB disease and no cultivar has been identified with strong HLB resistance. Citrus canker is also an economically important disease associated with a bacterial pathogen (Xanthomonas citri). In this study, we characterized endogenous citrus thionins and investigated their expression in different citrus tissues. Since no HLB-resistant citrus cultivars have been identified, we attempted to develop citrus resistant to both HLB and citrus canker through overexpression of a modified plant thionin. To improve effectiveness for disease resistance, we modified and synthesized the sequence encoding a plant thionin and cloned into the binary vector pBinPlus/ARS. The construct was then introduced into Agrobacterium strain EHA105 for citrus transformation. Transgenic Carrizo plants expressing the modified plant thionin were generated by Agrobacterium-mediated transformation. Successful transformation and transgene gene expression was confirmed by molecular analysis. Transgenic Carrizo plants expressing the modified thionin gene were challenged with X. citri 3213 at a range of concentrations, and a significant reduction in canker symptoms and a decrease in bacterial growth were demonstrated compared to nontransgenic plants. Furthermore, the transgenic citrus plants were challenged with HLB via graft inoculation. Our results showed significant Las titer reduction in roots of transgenic Carrizo compared with control plants and reduced scion Las titer 12 months after graft inoculation. These data provide promise for engineering citrus disease resistance against HLB and canker. PMID:27499757

  7. Induced mutations in citrus

    International Nuclear Information System (INIS)

    Full text: Parthenocarpic tendency is an important prerequisite for successful induction of seedlessness in breeding and especially in mutation breeding. A gene for asynapsis and accompanying seedless fruit has been found by us in inbred progeny of cv. 'Wilking'. Using budwood irradiation by gamma rays, seedless mutants of 'Eureka' and 'Villafranca' lemon (original clone of the latter has 25 seeds) and 'Minneola' tangelo have been obtained. Ovule sterility of the three mutants is nearly complete, with some pollen fertility still remaining. A semi-compact mutant of Shamouti orange has been obtained by irradiation. A programme for inducing seedlessness in easy peeling citrus varieties and selections has been initiated. (author)

  8. Antihyperlipidemic effects of Citrus sinensis, Citrus paradisi, and their combinations

    OpenAIRE

    Neelam Mallick; Rafeeq Alam Khan

    2016-01-01

    Objective: Hyperlipidemia, extensively contributes in the progression of coronary heart diseases and atherosclerosis, but may be managed through alterations in the nutritional pattern. Several studies show that diet rich in polyphenols and antioxidants have antiatherogenic effects. Citrus sinensis and Citrus paradisi are widely known for health benefits and have found to produce antioxidant, anti-inflammatory, and hypolipidemic effects, hence current research was planned to determine the hypo...

  9. Comprehensive meta-analysis, co-expression, and miRNA nested network analysis identifies gene candidates in citrus against Huanglongbing disease

    OpenAIRE

    Rawat, Nidhi; Kiran, Sandhya P.; Du, Dongliang; Gmitter, Fred G; Deng, Zhanao

    2015-01-01

    Background Huanglongbing (HLB), the most devastating disease of citrus, is associated with infection by Candidatus Liberibacter asiaticus (CaLas) and is vectored by the Asian citrus psyllid (ACP). Recently, the molecular basis of citrus–HLB interactions has been examined using transcriptome analyses, and these analyses have identified many probe sets and pathways modulated by CaLas infection among different citrus cultivars. However, lack of consistency among reported findings indicates that ...

  10. Citrus Canker and Citrus Huanglongbing, Two Exotic Bacterial Diseases Threatening the Citrus Industries of the Western Hemisphere

    Science.gov (United States)

    Two exotic Asian bacterial diseases of citrus are currently plaguing citrus industries in the Western Hemisphere. The two largest citrus producing areas in the Americas, located in Florida and the state of São Paulo Brazil, are presently battling these devastating diseases. The presence of these d...

  11. CITRUS CANKER: PLANT PATHOLOGY VERSUS PUBLIC POLICY

    Science.gov (United States)

    Increasing international travel and trade has resulted in an unprecedented number of plant pathogen introductions, including Xanthomonas axonopodis pv citri, (Xac), the bacterium that causes citrus canker. The disease affects commercial and dooryard citrus, and has far-reaching politi...

  12. Comparative transcripts profiling reveals new insight into molecular processes regulating lycopene accumulation in a sweet orange (Citrus sinensis red-flesh mutant

    Directory of Open Access Journals (Sweden)

    Zhang Jianchen

    2009-11-01

    Full Text Available Abstract Background Interest in lycopene metabolism and regulation is growing rapidly because accumulative studies have suggested an important role for lycopene in human health promotion. However, little is known about the molecular processes regulating lycopene accumulation in fruits other than tomato so far. Results On a spontaneous sweet orange bud mutant with abnormal lycopene accumulation in fruits and its wild type, comparative transcripts profiling was performed using Massively Parallel Signature Sequencing (MPSS. A total of 6,877,027 and 6,275,309 reliable signatures were obtained for the wild type (WT and the mutant (MT, respectively. Interpretation of the MPSS signatures revealed that the total number of transcribed gene in MT is 18,106, larger than that in WT 17,670, suggesting that newly initiated transcription occurs in the MT. Further comparison of the transcripts abundance between MT and WT revealed that 3,738 genes show more than two fold expression difference, and 582 genes are up- or down-regulated at 0.05% significance level by more than three fold difference. Functional assignments of the differentially expressed genes indicated that 26 reliable metabolic pathways are altered in the mutant; the most noticeable ones are carotenoid biosynthesis, photosynthesis, and citrate cycle. These data suggest that enhanced photosynthesis and partial impairment of lycopene downstream flux are critical for the formation of lycopene accumulation trait in the mutant. Conclusion This study provided a global picture of the gene expression changes in a sweet orange red-flesh mutant as compared to the wild type. Interpretation of the differentially expressed genes revealed new insight into the molecular processes regulating lycopene accumulation in the sweet orange red-flesh mutant.

  13. CIBMAN: Database exploring Citrus biodiversity of Manipur

    OpenAIRE

    Sanabam, Rakesh; Somkuwar, Bharat Gopalrao; Thingnam, Gourshyam; Moirangthem, Sila; Handique, Pratap Jyoti; Huidrom, Sunitibala

    2012-01-01

    The rich wealth of Citrus genetic resources makes India to enjoy a remarkable position in the “Citrus belt of the world”. We have developed CIBMAN, a unique database on Citrus biodiversity of Manipur which comprises 33 accessions collected through extensive survey for more than three years. CIBMAN provides integrated access to Citrus species through sophisticated web interface which has following capabilities a) morphological details, b) socio-economic details, c) taxonomic details and d) geo...

  14. Citrus tristeza virus-host interactions

    OpenAIRE

    Dawson, William O.; Garnsey, Stephen M.; Satyanarayana eTatineni; Folimonova, Svetlana Y.; Scott J Harper; S eGowda

    2013-01-01

    Citrus tristeza virus (CTV) is a phloem-limited virus whose natural host range is restricted to citrus and related species. Although the virus has killed millions of trees, almost destroying whole industries, and continually limits production in many citrus growing areas, most isolates are mild or symptomless in most of their host range. There is little understanding of how the virus causes severe disease in some citrus and none in others. Movement and distribution of CTV differs considerably...

  15. Citrus breeding, genetics and genomics in Japan

    OpenAIRE

    Omura, Mitsuo; Shimada, Takehiko

    2016-01-01

    Citrus is one of the most cultivated fruits in the world, and satsuma mandarin (Citrus unshiu Marc.) is a major cultivated citrus in Japan. Many excellent cultivars derived from satsuma mandarin have been released through the improvement of mandarins using a conventional breeding method. The citrus breeding program is a lengthy process owing to the long juvenility, and it is predicted that marker-assisted selection (MAS) will overcome the obstacle and improve the efficiency of conventional br...

  16. Soil Micronutrients and Citrus Growth

    Institute of Scientific and Technical Information of China (English)

    OUYANGTAO

    1993-01-01

    By using nutritional diagnosis of citrus leaves and determining soil micronutrients,the relationship between soil micronutrients and citrus growth in southern China has been studied.Studies showed that there was a significant positive correlation between available micronutrients (such as Zn,Mo,Cu)in the soil and the corresponding nutrients in citrus leaves.Thus,one can roughly learn of the sufficiency or deficiency of certain nutrients in soils by analyzing citrus leaves.Rational spray of Zn B or Mo fertilizer not only improved citrus yields but also increased the total sugar of Satsuma mandarin and of Xinhui orange by 2.9 and 17.2% respectively compared with the control.Spraying Mo fertilizer increased the vitamin C content of Satsuma mandarin juice by 4.7%-8.4%,maturated fruits 7-10 days earlier and gave the peel a brighter color.The ultramicroscopic characteristics of Zn-deficient citrus leaves were investigated under an electron microscope.Results showed that the Zn-deficient leaf cell was characterized mainly by poor cytoplasm,endoplasmic reticula and ribosomes and by big starch grains in the chloroplast.As a result of spraying Zn fertilizer the structure of the cell returned to normal,the cytoplasm became rich and the amount of chloroplast increased.There also appeared a great deal of multiform endoplasmic reticula,thus promoting the photosynthesis of Zn-deficient plants.This provides a cytologico-theoretical basis for fertilization of high-yielding citrus trees.

  17. Citrus Limonoids: Analysis, Bioactivity, and Biomedical Prospects

    Science.gov (United States)

    This publication is a review of the chemistry, biochemistry and bioactivity of limonoids occurring in citrus. The review chronologically relates the evolution of research in citrus limonoids beginning with their association with bitterness development in citrus juices. The chemical and biochemical...

  18. A stable RNA virus-based vector for citrus trees

    International Nuclear Information System (INIS)

    Virus-based vectors are important tools in plant molecular biology and plant genomics. A number of vectors based on viruses that infect herbaceous plants are in use for expression or silencing of genes in plants as well as screening unknown sequences for function. Yet there is a need for useful virus-based vectors for woody plants, which demand much greater stability because of the longer time required for systemic infection and analysis. We examined several strategies to develop a Citrus tristeza virus (CTV)-based vector for transient expression of foreign genes in citrus trees using a green fluorescent protein (GFP) as a reporter. These strategies included substitution of the p13 open reading frame (ORF) by the ORF of GFP, construction of a self-processing fusion of GFP in-frame with the major coat protein (CP), or expression of the GFP ORF as an extra gene from a subgenomic (sg) mRNA controlled either by a duplicated CTV CP sgRNA controller element (CE) or an introduced heterologous CE of Beet yellows virus. Engineered vector constructs were examined for replication, encapsidation, GFP expression during multiple passages in protoplasts, and for their ability to infect, move, express GFP, and be maintained in citrus plants. The most successful vectors based on the 'add-a-gene' strategy have been unusually stable, continuing to produce GFP fluorescence after more than 4 years in citrus trees

  19. Preliminary molecular basis of Danggui-buxue-tang on Qi deficiency and blood stasis syndrome

    Institute of Scientific and Technical Information of China (English)

    LIU Ya; LI Xiao-hui

    2008-01-01

    Objective To study the molecular biology mechanism of Danggui-buxue-tang on tonifying Qi and controlling blood circulation through modulation the immune functional genes. Methods Rats were randomly divided into control group, model group, and Danggui-buxue-tang group. Effect of Dang-guibuxue-tang on mRNA expressions of IL-1β, TNF-α, HSP-70, NF-kB, p38MAPK and JNK in blood cells of rats with Qi deficiency and blood stasis were measured with real-time fluorescent quantitative-PCR at 5th, 14th and 28th day. And that in artery wall were determined at the 28th day. NF-kB/p65 and p-c-jun protein expressions in rat artery wall were detected by western blotting as well. Results In model group, TNF-α, IL-1β, NF-kB, HSP-70, p38MAPK and JNK mRNA expression in blood cell increased significantly compared with control group. Compared with model group, mRNA expression of IL-1βand JNK decreased significantly; TNF-α decreased at 5th, 14th day;HSP-70, p38MAPK decreased at 14th, 28th day; NF-kB decreased at 28th day. In model group, TNF-α, IL-1β, NF-kB, p38MAPK and JNK mRNA expression in artery increased compared with control group, excluded HSP-70, and that in Danggui-buxue-tang group decreased significantly. In model group, NF-kB/p65 and p-c-jun protein expression in artery increased compared with control group, and that in Danggui-buxue-tang group decreased significantly. Conclusions The effects of Dang-guibuxuetang on Qi deficiency and blood stasis syndrome was brought from the regulating of cytokine network at multi-link and multi-target, NF-kB, p38MAPK and JNK signal transduction pathways included. Through which the immune system and whole body reached to a functional balance status.

  20. Phytochemical, antimicrobial, and antioxidant activities of different citrus juice concentrates

    OpenAIRE

    Oikeh, Ehigbai I.; Omoregie, Ehimwenma S; Oviasogie, Faith E.; Oriakhi, Kelly

    2015-01-01

    Abstract The search for new antimicrobial compounds is ongoing. Its importance cannot be overemphasized in an era of emerging resistant pathogenic organisms. This study therefore investigated the phytochemical composition and antioxidant and antimicrobial activities of different citrus juice concentrates. Fruit juices of Citrus tangerine (tangerine), Citrus paradisi (grape), Citrus limon (lemon), and Citrus aurantifolia (lime) were evaluated. Antimicrobial activities against five bacterial an...

  1. Molecular biomarkers of Dreissena polymorpha for evaluation of renaturation success of a formerly sewage polluted stream

    Energy Technology Data Exchange (ETDEWEB)

    Contardo-Jara, Valeska [Leibniz-Institute of Freshwater Ecology and Inland Fisheries, Department of Inland Fisheries, Biochemical Regulation, Mueggelseedamm 301, 12587 Berlin (Germany)], E-mail: contardo@igb-berlin.de; Wiegand, Claudia [Leibniz-Institute of Freshwater Ecology and Inland Fisheries, Department of Inland Fisheries, Biochemical Regulation, Mueggelseedamm 301, 12587 Berlin (Germany); Humboldt University Berlin, Faculty of Biology, Unter den Linden 6, 10099 Berlin (Germany)], E-mail: cwiegand@igb-berlin.de

    2008-09-15

    The renaturation success of an urban stream, formally used for discharge of treated sewage waters was investigated by active biomonitoring with Dreissena polymorpha based on molecular biomarkers and compared to a semi-natural stream and laboratory controls. Response to pollution charges were analyzed by reverse transcriptase-PCR of heat-shock protein (hsp70), P-glycoprotein (P-gp), catalase (CAT) and pi class glutathione S-transferase (piGST). Hsp70 transcription was similarly induced at both sites, indicating protein damage. At the semi-natural stream CAT and P-gp were induced, indicating oxidative stress and increased discharge of pollutants, which correlated to high amounts of aluminum at this site. piGST was induced at one sampling date at the renaturated stream only, but identification of the causing pollutant was not achieved. Results confirm regeneration of the formerly sewage polluted stream, because induction of the tested biomarkers was either at or below the levels of the semi-natural stream. - A new application for environmental health assessment combining the use of species tolerant to pollution, Dreissena polymorpha, and molecular tools (RT-PCR) in active biomonitoring.

  2. Molecular biomarkers of Dreissena polymorpha for evaluation of renaturation success of a formerly sewage polluted stream

    International Nuclear Information System (INIS)

    The renaturation success of an urban stream, formally used for discharge of treated sewage waters was investigated by active biomonitoring with Dreissena polymorpha based on molecular biomarkers and compared to a semi-natural stream and laboratory controls. Response to pollution charges were analyzed by reverse transcriptase-PCR of heat-shock protein (hsp70), P-glycoprotein (P-gp), catalase (CAT) and pi class glutathione S-transferase (piGST). Hsp70 transcription was similarly induced at both sites, indicating protein damage. At the semi-natural stream CAT and P-gp were induced, indicating oxidative stress and increased discharge of pollutants, which correlated to high amounts of aluminum at this site. piGST was induced at one sampling date at the renaturated stream only, but identification of the causing pollutant was not achieved. Results confirm regeneration of the formerly sewage polluted stream, because induction of the tested biomarkers was either at or below the levels of the semi-natural stream. - A new application for environmental health assessment combining the use of species tolerant to pollution, Dreissena polymorpha, and molecular tools (RT-PCR) in active biomonitoring

  3. Molecular docking and molecular dynamics studies reveal structural basis of inhibition and selectivity of inhibitors EGCG and OSU-03012 toward glucose regulated protein-78 (GRP78) overexpressed in glioblastoma.

    Science.gov (United States)

    Bhattacharjee, Rituparna; Devi, Arpita; Mishra, Seema

    2015-10-01

    Glioblastoma (GBM), a malignant form of brain tumor, has a high mortality rate. GRP78, one of the HSP70 protein family members, is overexpressed in GBM. GRP78 is the key chaperone protein involved in the unfolded protein response. Upregulated GRP78 expression in cancer cells inhibits apoptosis and promotes chemoresistance. GRP78 has an ATPase domain, a substrate-binding domain, and a linker region. ATP-competitive inhibitors such as EGCG and OSU-03012 inhibit GRP78 activity and reduce its expression in GBM. However, there is a lack of structural data on the binding modes of these inhibitors to GRP78 ATPase domain. Further, the mode of selectivity of these inhibitors toward GRP78 also is unknown. Toward this end, molecular docking was performed with AutoDock Vina and confirmation obtained by docking using ROSIE. The stability and MM-PBSA binding energy of GRP78-inhibitor complexes as well as energetic contribution of individual residues was analyzed by 50 ns molecular dynamics run with GROMACS. MSA by ClustalW2 identified unique amino acid residues in the ATPase domain of GRP78 which were different from the residues present in other HSP70 proteins. Important and unique amino acid residues of GRP78 such as Ile61, Glu293, Arg297, and Arg367 played a major role in the intermolecular interactions with these inhibitors. The interactions with unique residues of GRP78 as compared with those of HSP70-1A provided the basis for selectivity. It was found that the binding affinity and specificity/selectivity of EGCG toward GRP78 was higher than that toward HSP70-1A, and selectivity was even better than OSU-03012. OSU-03012 was predicted to bind to GRP78. Analyses from MD runs showed tight binding and stability of complexes, and the highest number of hydrogen bonds during the trajectory runs were comparable to those found in the docking studies. Energetic contribution of individual inhibitor-interacting residues showed that energy values of Ile61 and Glu293 were among the most

  4. Effectivity of Kaffir lime (Citrus hystrix, Nasnaran Mandarin (Citrus amblycarpa, and Pomelo (Citrus maxima Leaf Extract Against Aedes aegypti Larvae

    Directory of Open Access Journals (Sweden)

    Hebert Adrianto

    2014-03-01

    Full Text Available The used of chemicals as larvicidal may causing resistance, health problem, and environment problem. Leaf extracts of Citrus as biolarvicidal against Aedes aegypti larvae can be used as an alternative larvicidal. The aims of this study is to find out the species of Citrus in the study sample, which is the most effective as biolarvicidal. This study was conducted using nested experiment design, with six treatments and five replicates. Larvae mortality was observed after 24 and 48 hours. Then, data were analyzed by probit. The results of this study show that (1 the LC95 value after 24 hours of exposure of leaf extracts of Citrus hystrix, Citrus amblycarpa, and Citrus maxima, were each 3,176 ppm; 4,174 ppm; and 6,369 ppm. (2 the LC95 value after 48 hours of exposure of leaf extracts of Citrus hystrix, Citrus amblycarpa, and Citrus maxima, were each 2,499 ppm; 3,256 ppm; and 4,886 ppm. (3 leaf extract of Citrus hystrix is the most effective among others as biolarvicidal against Aedes aegypti larvae. Leaf extract of Citrus hystrix can be used as alternative biolarvicidal.

  5. Establishment and Application of SCoT Molecular Marker System for Citrus%柑橘SCoT分子标记技术体系的建立及其在遗传分析中的应用

    Institute of Scientific and Technical Information of China (English)

    韩国辉; 向素琼; 汪卫星; 贾志刚; 洪棋斌; 梁国鲁

    2011-01-01

    采用正交设计方法,对影响柑橘SCoT-PCR反应的Mg2+、dNTPs、引物、Taq DNA聚合酶及模板DNA用量等因素进行优化,建立了适于柑橘的SCoT标记分析体系。20μL反应体系中含有:Mg2+1.5 mmol.L-1,dNTPs 0.35 mmol.L-1,引物0.625μmol.L-1,TaqDNA聚合酶0.25 U,模板DNA 10 ng。最适退火温度为49℃。利用该体系对Wan2橘橙×Li2甜橙的杂交后代及沙田柚四倍体进行分析,扩增条带清晰,扩增产物在150~2 100 bp之间。7个引物在%Orthogonal design was applied to optimize SCoT-PCR amplification system of Citrus in five factors such as Mg2+,dNTPs,primer,Taq DNA polymerase and template DNA.An optimal reaction system of Citrus was established,PCR reaction mixtures contained Mg2+ 1.5 mmol.L-1,dNTPs 0.35 mmol.L-1,primers 0.625 μmol.L-1,Taq polymerase 0.25 U,template DNA 10 ng.The most suitableannealing temperature of primers was 49 ℃.Amplifications and genetic analysis were carried out on four tetraploids of Shatianyou pomelo [Citrus grandis(L.)Osb.‘Shatianyou’]and progenies from Wan2 [tangor,Citrus unshiu(Mark.)Marc.× Citrus sinensis(L.)Osb.] × Li2[sweet orange,Citrus sinensis(L.)Osb.].The size range was typically between 150–2 100 bp.The 7 primers generated a total of 74fragments from Wan2,Li2 and their progenies,48(64.9%)were polymorphic,and 18 progenies could be grouped into five distinct families based on similarity of 0.85,which indicated that greater genetic diversity was existence in the hybrids.And 84 bands were amplified from Shatianyou pomelo with 11 primers,of which 46(54.8%)were polymorphic.UPGMA cluster analysis showed that the similarity coefficient of five accessions ranged from 0.785 to 0.880.Four tetraploids could be grouped into three distinct families based on similarity of 0.825,and three homologous tetraploids were clustered in different locations,which showed that different degrees of genome variation was detected in tetraploids compared with

  6. Comparison of different diagnostic methods for detection of Hop stunt viroid and Citrus exocortis viroid in citrus

    Directory of Open Access Journals (Sweden)

    Cecilia Escobar Ponce de León

    2012-06-01

    Full Text Available Viroids such as Hop stunt viroid (HSVd and Citrus exocortis viroid (CEVd are important pathogens in citrus plants. A comparative assay was performed to evaluate the viroid diagnostic techniques currently available at the Citrus Sanitation Center of Estación Experimental Agroindustrial Obispo Colombres. We compared the results obtained in the diagnosis of HSVd and CEVd in citrus using two molecular techniques: s-PAGE and RT-PCR and biological indexing. Biological indexing was performed in a greenhouse under controlled temperature, using Etrog citron Arizona 861-S1 grafted on rough lemon rootstock as the indicator plant. Tissue samples for s-PAGE were obtained from inoculated citrons used in the biological indexing tests, and DNA extraction was performed following published procedures. HSVd and CEVd specific primers were used for RT-PCR, and samples were taken directly from field plants and inoculated citrons. Two nucleic acid extraction methods were compared. A total of 12 viroid isolates collected from orange, citron, grapefruit, citrumelo and lime trees, mostly from North Western Argentina, were used in these assays. The three diagnostic methods compared gave similar results in 11 of 12 samples assayed. Several viroids were detected by s-PAGE and most of them were found as mixtures. HSVd and CEVd were identified by RT-PCR and no differences were found between the two extraction methods assayed. Sample R-407 was positive for exocortis with biological diagnosis and s-PAGE, but it was negative with RT-PCR. This was probably due to inefficient cDNA synthesis or to the presence of inhibitors that could interfere with the reaction. On the basis of the results obtained, it can be concluded that the three techniques assayed are complementary and useful tools for diagnosis and identification of HSVd and CEVd in citrus.

  7. Isolation, characterization and modification of citrus pectins

    Directory of Open Access Journals (Sweden)

    MARIA KRATCHANOVA

    2012-01-01

    Full Text Available Orange and lemon peels were used for obtaining pectic polysaccharides. Citrus peels were previously treated with 96% ethanol, and the obtained alcohol-insoluble solids (AIS were subjected to a sequential extraction with hot distilled water and hot 0.5% HCl. Water- and acid-extracted orange (WEOP and AEOP and lemon (WELP and AELP pectins were obtained. Acid extraction gave higher yields of pectin than water extraction and lemon peels were richer in pectin. Comparative investigations were carried out with chromatographically purified commercial citrus pectin (CPCP. Chemical and physicochemical characterization of all pectins was accomplished. It was found that pectins were similar in anhydrouronic acid content (AUАC, 69-81%, but differed in their degree of methylesterification (DM, 55-81%. Generally water-extracted pectins were with higher DM. Both orange pectins were with higher DM and degree of acetylation (DA, 2%, in comparison with the corresponding lemon pectins. Water-extracted pectins were with higher degree of feruloylation (DF, 0.12-0.34%. To our knowledge this is the first report on the estimation of ester-linked ferulic acid in orange and lemon peel pectins. Pectic polysaccharides differed in molecular weight and homogeneity. WELP was with the highest molecular weight and homogeneity. The pectins contained D-galacturonic and D-glucuronic acids, L-arabinose, D-galactose, L-fucose, L-rhamnose and D-xylose. All investigated pectins showed immunostimulating activity by complement activation in the classical pathway at 1.25 and 2.5 mg/mL. Pectic polysaccharides were modified with endopolygalacturonase. Enzyme-modified CPCP and WEOP had higher anti-complementary activity than the corresponding initial pectins.

  8. 78 FR 8435 - Importation of Fresh Citrus Fruit From Uruguay, Including Citrus Hybrids and Fortunella

    Science.gov (United States)

    2013-02-06

    ... Animal and Plant Health Inspection Service 7 CFR Part 319 RIN 0579-AD59 Importation of Fresh Citrus Fruit From Uruguay, Including Citrus Hybrids and Fortunella spp., Into the Continental United States AGENCY.... australis. \\2\\...

  9. 76 FR 23449 - Citrus Canker, Citrus Greening, and Asian Citrus Psyllid; Interstate Movement of Regulated...

    Science.gov (United States)

    2011-04-27

    ... 22, 2007 and effective on March 16, 2007 (72 FR 13423-13428, Docket No. APHIS-2007-0032) that....aphis.usda.gov/plant_health/plant_pest_info/citrus/index.shtml . We believe that the procedures set... effective on June 17, 2010 (75 FR 34322-34336, Docket No. APHIS- 2008-0015).\\4\\ Several commenters on...

  10. Identification and Characterization of Seven Glutathione S-Transferase Genes from Citrus Red Mite, Panonychus citri (McGregor)

    OpenAIRE

    Liao, Chong-Yu; Zhang, Kun; Niu, Jin-Zhi; Ding, Tian-Bo; Zhong, Rui; Xia, Wen-Kai; Dou, Wei; Wang, Jin-Jun

    2013-01-01

    The citrus red mite, Panonychus citri (McGregor), is a global citrus pest, and has developed severe resistance to several types of acaricides. However, the molecular mechanisms of resistance in this mite remain unknown. In this study, seven full-length cDNAs encoding glutathione S-transferases (GSTs) genes were identified and characterized in P. citri. The effects of pyridaben and fenpropathrin exposure on the expression of these genes were also investigated. Phylogenetic analysis revealed th...

  11. Analysis of Transcriptome Differences between Resistant and Susceptible Strains of the Citrus Red Mite Panonychus citri (Acari: Tetranychidae)

    OpenAIRE

    Bin Liu; Gaofei Jiang; Yunfei Zhang; Junli Li; Xiaojiao Li; Jiansu Yue; Fei Chen; Haoqiang Liu; Hongjun Li; Shiping Zhu; Jinjun Wang; Chun Ran

    2011-01-01

    BACKGROUND: The citrus red mite is a worldwide citrus pest and a common sensitizing allergen of asthma and rhinitis. It has developed strong resistance to many registered acaricides, However, the molecular mechanisms of resistance remain unknown. we therefore used next generation sequencing technology to investigate the global transcriptomes between resistant strains and susceptible strains. RESULTS: We obtained 34,159, 30,466 and 32,217 unigenes by assembling the SS reads, RS reads and SS&RS...

  12. Community Analysis of Arbuscular Mycorrhizal Fungi in Roots of Poncirus trifoliata and Citrus reticulata Based on SSU rDNA

    OpenAIRE

    Wang, Peng; Wang, Yin

    2014-01-01

    Morphological observation of arbuscular mycorrhizal fungi (AMF) species in rhizospheric soil could not accurately reflect the actual AMF colonizing status in roots, while molecular identification of indigenous AMF colonizing citrus rootstocks at present was rare in China. In our study, community of AMF colonizing trifoliate orange (Poncirus trifoliata L. Raf.) and red tangerine (Citrus reticulata Blanco) were analyzed based on small subunit of ribosomal DNA genes. Morphological observation sh...

  13. 75 FR 34322 - Citrus Greening and Asian Citrus Psyllid; Quarantine and Interstate Movement Regulations

    Science.gov (United States)

    2010-06-17

    ... Animal and Plant Health Inspection Service 7 CFR Parts 301 and 305 RIN 0579-AC85 Citrus Greening and Asian Citrus Psyllid; Quarantine and Interstate Movement Regulations AGENCY: Animal and Plant Health... African citrus psyllid is not present in the United States. The Animal and Plant Health Inspection...

  14. Quantitative Distribution of Candidatus Liberibacter asiaticus in Citrus Plants and Fruits Infected by Citrus Huanglongbing

    Science.gov (United States)

    Citrus huanglongbing (HLB), or greening disease, is caused by any of three non culturable Gram-negative bacteria belonging to Candidatus Liberibacter spp. The pathogens are transmitted by citrus psyllids to all commercial varieties of citrus. The diseases are lethal and have recently become widespr...

  15. Antimyeloma Effects of the Heat Shock Protein 70 Molecular Chaperone Inhibitor MAL3-101

    Directory of Open Access Journals (Sweden)

    Marc J. Braunstein

    2011-01-01

    Full Text Available Multiple myeloma (MM is the second most common hematologic malignancy and remains incurable, primarily due to the treatment-refractory/resistant nature of the disease. A rational approach to this compelling challenge is to develop new drugs that act synergistically with existing effective agents. This approach will reduce drug concentrations, avoid treatment resistance, and also improve treatment effectiveness by targeting new and nonredundant pathways in MM. Toward this goal, we examined the antimyeloma effects of MAL3-101, a member of a new class of non-ATP-site inhibitors of the heat shock protein (Hsp 70 molecular chaperone. We discovered that MAL3-101 exhibited antimyeloma effects on MM cell lines in vitro and in vivo in a xenograft plasmacytoma model, as well as on primary tumor cells and bone marrow endothelial cells from myeloma patients. In combination with a proteasome inhibitor, MAL3-101 significantly potentiated the in vitro and in vivo antimyeloma effects. These data support a preclinical rationale for small molecule inhibition of Hsp70 function, either alone or in combination with other agents, as an effective therapeutic strategy for MM.

  16. Signaling pathways in a Citrus EST database

    OpenAIRE

    Angela Mehta; Marilia Santos Silva; Simone Guidetti-Gonzalez; Helaine Carrer; Marco Aurélio Takita; Natália F. Martins

    2007-01-01

    Citrus spp. are economically important crops, which in Brazil are grown mainly in the State of São Paulo. Citrus cultures are attacked by several pathogens, causing severe yield losses. In order to better understand this culture, the Millenium Project (IAC Cordeirópolis) was launched in order to sequence Citrus ESTs (expressed sequence tags) from different tissues, including leaf, bark, fruit, root and flower. Plants were submitted to biotic and abiotic stresses and investigated under differe...

  17. Recent advances in Citrus psorosis virus

    OpenAIRE

    Achachi, Asmae; Ait Barka, Essaïd; Ibriz, Mohammed

    2014-01-01

    Psorosis is a globally devastating disease of citrus caused by an infectious filamentous ophiovirus, Citrus psorosis virus (CPsV), which causes annual losses of about 5 % and a progressive decline of trees by affecting the conductive tissues. The disease can be harboured asymptomatically in many citrus species. In the field, the most characteristic symptoms of the disease in adult trees are bark scaling in the trunk and main branches and also internal staining in the underlying wood. The viru...

  18. Antimicrobial Compounds to Combat Citrus Huanglongbing

    OpenAIRE

    Zhang, Muqing; Guo, Ying; Powell, Charles A.; Duan, Yongping

    2014-01-01

    Citrus Huanglongbing (HLB) is associated with the fastidious bacterium,Candidatus Liberibacter, (Las) that is transmitted by a phloem-feeding insect (Citrus Psyllid). An ideal solution to combat citrus HLB is to completely eliminate the bacteria after a single course of the chemotherapy, either active directly on the bacteria or indirectly through induction of host defense compounds. Twenty-seven antimicrobial compounds were screened to test for in vivo activities against HLB bacterium while ...

  19. The aconitate hydratase family from Citrus

    OpenAIRE

    Cercos Manuel; Talon Manuel; Soler Guillermo; Terol Javier

    2010-01-01

    Abstract Background Research on citrus fruit ripening has received considerable attention because of the importance of citrus fruits for the human diet. Organic acids are among the main determinants of taste and organoleptic quality of fruits and hence the control of fruit acidity loss has a strong economical relevance. In citrus, organic acids accumulate in the juice sac cells of developing fruits and are catabolized thereafter during ripening. Aconitase, that transforms citrate to isocitrat...

  20. Green synthesis of gold nanoparticles using Citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) aqueous extract and its characterization

    Science.gov (United States)

    Sujitha, Mohanan V.; Kannan, Soundarapandian

    2013-02-01

    This study reports the biological synthesis of gold nanoparticles by the reduction of HAuCl4 by using citrus fruits (Citrus limon, Citrus reticulata and Citrus sinensis) juice extract as the reducing and stabilizing agent. A various shape and size of gold nanoparticles were formed when the ratio of the reactants were altered with respect to 1.0 mM chloroauric acid solution. The gold nanoparticles obtained were characterized by UV-visible spectra, transmission electron microscopy (TEM) and X-ray diffraction (XRD). TEM studies showed the particles to be of various shapes and sizes and particle size ranges from 15 to 80 nm. Selected-area electron diffraction (SAED) pattern confirmed fcc phase and crystallinity of the particles. The X-ray diffraction analysis revealed the distinctive facets (1 1 1, 2 0 0, 2 2 0 and 2 2 2 planes) of gold nanoparticles. Dynamic light scattering (DLS) studies revealed that the average size for colloid gp3 of C. limon, C. reticulata and C. sinensis are 32.2 nm, 43.4 nm and 56.7 nm respectively. The DLS graph showed that the particles size was larger and more polydispersed compared to the one observed by TEM due to the fact that the measured size also includes the bio-organic compounds enveloping the core of the Au NPs. Zeta potential value for gold nanoparticles obtained from colloid gp3 of C. limon, C. reticulata and C. sinensis are -45.9, -37.9 and -31.4 respectively indicating the stability of the synthesized nanoparticles. Herein we propose a novel, previously unexploited method for the biological syntheses of polymorphic gold nanoparticles with potent biological applications.

  1. Influence of mitochondria on gene expression in a citrus cybrid.

    Science.gov (United States)

    Bassene, Jean-Baptiste; Froelicher, Yann; Navarro, Luis; Ollitrault, Patrick; Ancillo, Gema

    2011-06-01

    The production of cybrids, combining nucleus of a species with alien cytoplasmic organelles, is a valuable method used for improvement of various crops. Several citrus cybrids have been created by somatic hybridization. These genotypes are interesting models to analyze the impact of cytoplasmic genome change on nuclear genome expression. Herein, we report genome-wide gene expression analysis in leaves of a citrus cybrid between C. reticulata cv 'Willowleaf mandarin' and C. limon cv 'Eureka lemon' compared with its lemon parent, using a Citrus 20K cDNA microarray. Molecular analysis showed that this cybrid possesses nuclear and chloroplast genomes of Eureka lemon plus mitochondria from Willowleaf mandarin and, therefore, can be considered as a lemon bearing foreign mitochondria. Mandarin mitochondria influenced the expression of a large set of lemon nuclear genes causing an over-expression of 480 of them and repression of 39 genes. Quantitative real-time RT-PCR further confirmed the credibility of microarray data. Genes over-expressed in cybrid leaves are predominantly attributed to the functional category "cellular protein metabolism" whereas in the down-regulated none functional category was enriched. Overall, mitochondria replacement affected different nuclear genes including particularly genes predicted to be involved in mitochondrial retrograde signaling. Mitochondria regulate all cell structures even chloroplast status. These results suggest that nuclear gene expression is modulated with respect to new information received from the foreign organelle, with the final objective to suit specific needs to ensure better cell physiological balance. PMID:21308470

  2. Deep-sequencing revealed Citrus bark cracking viroid (CBCVd) as a highly aggressive pathogen on hop

    Czech Academy of Sciences Publication Activity Database

    Jakše, J.; Radišek, S.; Pokorn, T.; Matoušek, Jaroslav; Javornik, B.

    2015-01-01

    Roč. 64, č. 4 (2015), s. 831-842. ISSN 0032-0862 R&D Projects: GA MŠk(CZ) LH14255 Institutional support: RVO:60077344 Keywords : Bioinformatic * Citrus bark cracking viroid * Hop * Next-generation sequencing Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.121, year: 2014

  3. Genetic differentiation and biology of Citrus Tristeza virus populations spreading in California

    Science.gov (United States)

    CTV strains were collected from more than 1500 CTV-infected trees in citrus groves in Tulare, Kern, Ventura, Riverside and San Diego Counties to assess molecular and biological properties of CTV strains currently in California. Tests included serology with MCA13, quantitative (q) real-time Reverse ...

  4. 7 CFR 319.56-38 - Citrus from Chile.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Citrus from Chile. 319.56-38 Section 319.56-38... from Chile. Clementines (Citrus reticulata Blanco var. Clementine), mandarins (Citrus reticulata Blanco), and tangerines (Citrus reticulata Blanco) may be imported into the United States from Chile,...

  5. Cryopreservation and Cryotherapy of Citrus Cultivars

    Science.gov (United States)

    Long-term conservation of Citrus clones can be accomplished by cryopreservation. Shoot tips will survive liquid nitrogen exposure and storage when appropriately desiccated and treated with cryoprotectant solutions. In our research, vegetative Citrus budwood is shipped from Riverside to Fort Collin...

  6. Molecular identification of the Cryptosporidium deer genotype in the Hokkaido sika deer (Cervus nippon yesoensis) in Hokkaido, Japan.

    Science.gov (United States)

    Kato, Satomi; Yanagawa, Yojiro; Matsuyama, Ryota; Suzuki, Masatsugu; Sugimoto, Chihiro

    2016-04-01

    The protozoan Cryptosporidium occurs in a wide range of animal species including many Cervidae species. Fecal samples collected from the Hokkaido sika deer (Cervus nippon yesoensis), a native deer of Hokkaido, in the central, western, and eastern areas of Hokkaido were examined by polymerase chain reaction (PCR) to detect infections with Cryptosporidium and for sequence analyses to reveal the molecular characteristics of the amplified DNA. DNA was extracted from 319 fecal samples and examined with PCR using primers for small-subunit ribosomal RNA (SSU-rRNA), actin, and 70-kDa heat shock protein (HSP70) gene loci. PCR-amplified fragments were sequenced and phylogenetic trees were created. In 319 fecal samples, 25 samples (7.8 %) were positive with SSU-rRNA PCR that were identified as the Cryptosporidium deer genotype. Among Cryptosporidium-positive samples, fawns showed higher prevalence (16.1 %) than yearlings (6.4 %) and adults (4.7 %). The result of Fisher's exact test showed a statistical significance in the prevalence of the Cryptosporidium deer genotype between fawn and other age groups. Sequence analyses with actin and HSP70 gene fragments confirmed the SSU-rRNA result, and there were no sequence diversities observed. The Cryptosporidium deer genotype appears to be the prevalent Cryptosporidium species in the wild sika deer in Hokkaido, Japan. PMID:26687968

  7. Comparative Studies of Various Phytonutrients in Citrus Fruits

    Directory of Open Access Journals (Sweden)

    S. Ali

    2014-06-01

    Full Text Available The current piece of research was undertaken to investigate the phytochemicals constituents such as carbohydrates, proteins, alkaloids, anthraquinone, glycosides, Saponin, glycosides, steroid, cyanogenetic glycosides, tannins, phlobotannins, saponins, terpenoids, cardiac glycosides, etc in the species of citrus genus (Citrus sinensis, Citrus reticulate Blanco, Citrus aurantium L. All these plants were collected locally. Qualitative analysis of these phytochemicals was performed. All three types of citrus are rich with phytochemicals. Quantitative screenings is in progress in our lab.

  8. Accumulation of the sesquiterpenes nootkatone and valencene by callus cultures of Citrus paradisi, Citrus limonia and Citrus aurantium.

    Science.gov (United States)

    Del Río, J A; Ortuño, A; Puig, D G; Iborra, J L; Sabater, F

    1991-10-01

    The production of the sesquiterpenes nootkatone and valencene by callus cultures of Citrus species is described. The levels of these compounds were examined by gas chromatography-mass spectrometry and their yields were compared with the amounts found in mature fruits. A simultaneous increase and decrease in the levels of nootkatone and valencene, respectively, were observed with the aging of callus cultures of Citrus paradisi. These results suggest that valencene might be a possible precursor of nootkatone in this species. The high level of nootkatone detected in 9-month-old callus cultures of Citrus paradisi might be associated with the corresponding cell morphological changes observed. PMID:24221735

  9. Citrus Improvement Using Mutation Techniques

    International Nuclear Information System (INIS)

    Citrus cultivar improvement is hampered by several biological factors inherent to most citrus species. Facultative apomixis, self and cross-incompatibility, long juvenility period, and high heterozygosis are some of the vast arrays of impediments faced by citrus breeders in conventional hybridization. Since oranges and grapefruits are highly polyembryonic, the production of enough numbers of zygotic offspring for selection of superior genotypes of these species is basically impossible; hence, most commercially important cultivars of these species have originated through natural or induced mutation. Star Ruby, a deep-red-fleshed grapefruit, was developed by irradiation of Hudson grapefruit seeds with thermal neutrons. Unlike Hudson, which contains over 50 seeds per fruit, Star Ruby is nearly seedless. Hensz irradiated buds of Ruby Red grapefruit with thermal neutrons and a tree that originated from one of the buds produced fruits three times redder than Ruby Red. It was named A and I-1-48. Ten trees were propagated from A and I -1-48, and out of one of the trees, a budsport mutation was found producing fruits five times redder than Ruby Red. Called Rio Red, it is currently the variety of choice for Texas and is known worldwide for its sweetness, red flesh and beautiful blush. Currently, 37 years after A and I -1-48 was first propagated, the trees are still producing several budsport mutations. So far, in the 2007/2008 season, more than 100 new mutations were obtained from a 100-tree block. In the mandarin group, the existence of several monoembryonic cultivars facilitates conventional breeding, but still, induced mutation is part of most mandarin breeding programmes, and proprietary, new seedless cultivars have been produced in the US, Italy, Israel and elsewhere. Seedless mandarins produced by the University of California Riverside include Dayse, Fairchild, Encore, and Nova. The USDA-ARS, U.S. Horticultural Research Laboratory in Florida released a seedless

  10. Signaling pathways in a Citrus EST database

    Directory of Open Access Journals (Sweden)

    Angela Mehta

    2007-01-01

    Full Text Available Citrus spp. are economically important crops, which in Brazil are grown mainly in the State of São Paulo. Citrus cultures are attacked by several pathogens, causing severe yield losses. In order to better understand this culture, the Millenium Project (IAC Cordeirópolis was launched in order to sequence Citrus ESTs (expressed sequence tags from different tissues, including leaf, bark, fruit, root and flower. Plants were submitted to biotic and abiotic stresses and investigated under different development stages (adult vs. juvenile. Several cDNA libraries were constructed and the sequences obtained formed the Citrus ESTs database with almost 200,000 sequences. Searches were performed in the Citrus database to investigate the presence of different signaling pathway components. Several of the genes involved in the signaling of sugar, calcium, cytokinin, plant hormones, inositol phosphate, MAPKinase and COP9 were found in the citrus genome and are discussed in this paper. The results obtained may indicate that similar mechanisms described in other plants, such as Arabidopsis, occur in citrus. Further experimental studies must be conducted in order to understand the different signaling pathways present.

  11. Yield and quality responses of citrus (Citrus reticulate) and tea (Podocarpus fleuryi Hickel.) to compound fertilizers*

    OpenAIRE

    Wang, Rui; Shi, Xue-gen; Wei, You-zhang; Yang, Xiao-e; Uoti, Juhani

    2006-01-01

    Experiments were carried out with citrus (Citrus reticulate) and tea (Podocarpus fleuryi Hickel.) to study the effects of compound fertilizers on their yields and quality. In the citrus experiment, application of compound fertilizers increased available P, K and Mg contents in soil but decreased alkali-hydrolyzable N contents in soil and N, P and K contents in leaves. In the tea experiment, application of compound fertilizers increased available P, K and Mg contents in soil and N, P, K and Mg...

  12. Breeding, genetic and genomic of citrus for disease resistance

    Directory of Open Access Journals (Sweden)

    Marcos A. Machado

    2011-10-01

    Full Text Available Although the citriculture is one of the most important economic activities in Brazil, it is based on a small number of varieties. This fact has contributed for the vulnerability of the culture regarding the phytosanitary problems. A higher number of varieties/genotypes with potential for commercial growing, either for the industry or fresh market, has been one of the main objectives of citrus breeding programs. The genetic breeding of citrus has improved, in the last decades, due to the possibility of an association between biotechnological tools and classical methods of breeding. The use of molecular markers for early selection of zygotic seedlings from controlled crosses resulted in the possibility of selection of a high number of new combination and, as a consequence, the establishment of a great number of hybrids in field experiments. The faster new tools are incorporated in the program, the faster is possibility to reach new genotypes that can be tested as a new variety. Good traits should be kept or incorporate, whereas bad traits have to be excluded or minimized in the new genotype. Scion and rootstock can not be considered separately, and graft compatibility, fruit quality and productivity are essential traits to be evaluated in the last stages of the program. The mapping of QTLs has favored breeding programs of several perennial species and in citrus it was possible to map several characteristics with qualitative and quantitative inheritance. The existence of linkage maps and QTLs already mapped, the development of EST and BAC library and the sequencing of the Citrus complete genome altogether make very demanding and urgent the exploration of such data to launch a wider genetic study of citrus. The rising of information on genome of several organisms has opened new approaches looking for integration between breeding, genetic and genome. Genome assisted selection (GAS involves more than gene or complete genome sequencing and is becoming

  13. Lflg22, a Pathogen-Associated Molecular Pattern (PAMP) of Candidatus Liberibacter asiaticus, initiated differential PAMP-Triggered Immunity (PTI) in Grapefruit and Sun Chu Sha

    OpenAIRE

    Shi, Qingchun; Febres, Vicente J; Khalaf, Abeer; Moore, Gloria A.

    2014-01-01

    ‘Duncan’ Grapefruit (Citrus paradisi Macf.) and ‘Sun Chu Sha’ mandarin (C. reticulalta Blanco) represent two citrus genotypes that have different levels of tolerance to citrus greening or huanglongbing (HLB), a bacterial disease caused by Candidatus Liberibacter sp. In this study, the response of the two genotypes to the conserved 22 amino acid domain of the Liberibacter flagellin (Lflg22), a Pathogen-Associated Molecular Pattern (PAMP), were compared. The expression levels of citrus defense-...

  14. Microbial Production of Pectin from Citrus Peel

    OpenAIRE

    Sakai, Takuo; Okushima, Minoru

    1980-01-01

    A new method for the production of pectin from citrus peel was developed. For this purpose, a microorganism which produces a protopectin-solubilizing enzyme was isolated and identified as a variety of Trichosporon penicillatum. The most suitable conditions for the pectin production were determined as follows. Citrus (Citrus unshiu) peel was suspended in water (1:2, wt/vol), the organism was added, and fermentation proceeded over 15 to 20 h at 30°C. During the fermentation, the pectin in the p...

  15. A survey of FLS2 genes from multiple citrus species identifies candidates for enhancing disease resistance to Xanthomonas citri ssp. citri.

    OpenAIRE

    Qingchun Shi; Febres, Vicente J.; Jones, Jeffrey B; MOORE, Gloria A.

    2016-01-01

    Pathogen-associated molecular patterns (PAMPs)-triggered immunity (PTI) is an important component of plant innate immunity. In a previous study, we showed that the PAMP flg22 from Xanthomonas citri ssp. citri (Xflg22), the causal agent of citrus canker, induced PTI in citrus, which correlated with the observed levels of canker resistance. Here, we identified and sequenced two bacterial flagellin/flg22 receptors (FLS2-1 and FLS2-2) from ‘Duncan’ grapefruit (Citrus paradisi, CpFLS2-1 and CpFLS2...

  16. Citrus PH5-like H+-ATPase genes: identification and transcript analysis to investigate their possible relationship with citrate accumulation in fruits

    OpenAIRE

    Shi, Cai-Yun; Song, Rui-Qin; Hu, Xiao-Mei; Liu, Xiao; Jin, Long-Fei; Liu, Yong-Zhong

    2015-01-01

    PH5 is a petunia gene that encodes a plasma membrane H+-ATPase and determines the vacuolar pH. The citrate content of fruit cell vacuoles influences citrus organoleptic qualities. Although citrus could have PH5-like homologs that are involved in citrate accumulation, the details are still unknown. In this study, extensive data-mining with the PH5 sequence and PCR amplification confirmed that there are at least eight PH5-like genes (CsPH1-8) in the citrus genome. CsPHs have a molecular mass of...

  17. Green Synthesis and Biological Activities of Gold Nanoparticles Functionalized with Citrus reticulata, Citrus aurantium, Citrus sinensis and Citrus grandis

    International Nuclear Information System (INIS)

    In the present study, gold nanoparticles (GNPs) were prepared at boiling temperature (90-95 degree C) by treating gold ions with Citrus fruit extracts. The effect of mixing ratios of the reactants and concentration of gold hydrochloride was studied. In the standardization process, 10/sup -3/ M solution of HAuCl/sub 4/.3H/sub 2/O was reacted with fruit extracts for half an hour at 90-95 degree C in different ratios. GNPs were characterized by UV-Vis spectroscopy (UV-Vis) and atomic force microscopy (AFM). Their stability was evaluated against varying pH solutions and volumes of sodium chloride along with metals and antibiotics sensing ability. The gold nanoparticles were tested for antibacterial and antifungal activities against various pathogenic strains. The UV-Vis spectra of gold nanoparticles gave surface plasmon resonance at about 540 nm while the AFM images revealed the particle size within the range of 70-100 nm. GNPs showed remarkable stability in varying pH solutions and salt volumes as well as high detection ability towards cobalt, copper, ceftriaxone and penicillin. Moreover, the GNPs possessed moderate antibacterial and good antifungal activity. These results concluded that the Citrus fruit extracts can be utilized for large scale synthesis of cost-effective nanoparticles which may have compatibility for biomedical and pharmaceutical applications. (author)

  18. Effectivity of Kaffir lime (Citrus hystrix), Nasnaran Mandarin (Citrus amblycarpa), and Pomelo (Citrus maxima) Leaf Extract Against Aedes aegypti Larvae

    OpenAIRE

    Hebert Adrianto; Subagyo Yotopranoto; Hamidah

    2014-01-01

    The used of chemicals as larvicidal may causing resistance, health problem, and environment problem. Leaf extracts of Citrus as biolarvicidal against Aedes aegypti larvae can be used as an alternative larvicidal. The aims of this study is to find out the species of Citrus in the study sample, which is the most effective as biolarvicidal. This study was conducted using nested experiment design, with six treatments and five replicates. Larvae mortality was observed after 24 and 48 hours. Then, ...

  19. Physiological and Molecular Response of Prorocentrum minimum to Tannic Acid: An Experimental Study to Evaluate the Feasibility of Using Tannic Acid in Controling the Red Tide in a Eutrophic Coastal Water.

    Science.gov (United States)

    Jeong, Byungkwan; Jeong, Eui-Suk; Malazarte, Jacqueline Martha; Sin, Yongsik

    2016-01-01

    Bioassay and gene expression experiments were conducted in order to evaluate the growth and physiology of Prorocentrum minimum isolated from a eutrophic coastal water in response to tannic acid. In the bioassay experiments, variations in abundance, chlorophyll (chl) a concentration, maximum fluorescence (in vivo Fm), and photosynthetic efficiency (Fv/Fm) were measured over the course of a seven-day incubation. Moreover, stress-related gene expression in both the control and an experimental (2.5 ppm TA treatment) group was observed for 24 h and 48 h. The molecular markers used in this study were the heat shock proteins (Hsp70 and Hsp90) and cyclophilin (CYP). The findings show that P. minimum can thrive and grow at low concentrations (<2.5 ppm) of tannic acid, and, above this concentration, cells begin to slow down development. In addition, TA concentration of 10 ppm halted photosynthetic activity. At the molecular level, treatment with tannic acid increased the expression of Hsp70, Hsp90, and CYP, and heat shock proteins are more upregulated than the cyclophilin gene. Exposure to tannic acid increased the expression of stress factors over time (48 h) by 10- to 27-fold the expression level of the control group. These results suggest that tannic acid can be used to control harmful algal blooms such as those containing P. minimum in eutrophic coastal waters. PMID:27187440

  20. Physiological and Molecular Response of Prorocentrum minimum to Tannic Acid: An Experimental Study to Evaluate the Feasibility of Using Tannic Acid in Controling the Red Tide in a Eutrophic Coastal Water

    Directory of Open Access Journals (Sweden)

    Byungkwan Jeong

    2016-05-01

    Full Text Available Bioassay and gene expression experiments were conducted in order to evaluate the growth and physiology of Prorocentrum minimum isolated from a eutrophic coastal water in response to tannic acid. In the bioassay experiments, variations in abundance, chlorophyll (chl a concentration, maximum fluorescence (in vivo Fm, and photosynthetic efficiency (Fv/Fm were measured over the course of a seven-day incubation. Moreover, stress-related gene expression in both the control and an experimental (2.5 ppm TA treatment group was observed for 24 h and 48 h. The molecular markers used in this study were the heat shock proteins (Hsp70 and Hsp90 and cyclophilin (CYP. The findings show that P. minimum can thrive and grow at low concentrations (<2.5 ppm of tannic acid, and, above this concentration, cells begin to slow down development. In addition, TA concentration of 10 ppm halted photosynthetic activity. At the molecular level, treatment with tannic acid increased the expression of Hsp70, Hsp90, and CYP, and heat shock proteins are more upregulated than the cyclophilin gene. Exposure to tannic acid increased the expression of stress factors over time (48 h by 10- to 27-fold the expression level of the control group. These results suggest that tannic acid can be used to control harmful algal blooms such as those containing P. minimum in eutrophic coastal waters.

  1. Physiological and Molecular Response of Prorocentrum minimum to Tannic Acid: An Experimental Study to Evaluate the Feasibility of Using Tannic Acid in Controling the Red Tide in a Eutrophic Coastal Water

    Science.gov (United States)

    Jeong, Byungkwan; Jeong, Eui-Suk; Malazarte, Jacqueline Martha; Sin, Yongsik

    2016-01-01

    Bioassay and gene expression experiments were conducted in order to evaluate the growth and physiology of Prorocentrum minimum isolated from a eutrophic coastal water in response to tannic acid. In the bioassay experiments, variations in abundance, chlorophyll (chl) a concentration, maximum fluorescence (in vivo Fm), and photosynthetic efficiency (Fv/Fm) were measured over the course of a seven-day incubation. Moreover, stress-related gene expression in both the control and an experimental (2.5 ppm TA treatment) group was observed for 24 h and 48 h. The molecular markers used in this study were the heat shock proteins (Hsp70 and Hsp90) and cyclophilin (CYP). The findings show that P. minimum can thrive and grow at low concentrations (<2.5 ppm) of tannic acid, and, above this concentration, cells begin to slow down development. In addition, TA concentration of 10 ppm halted photosynthetic activity. At the molecular level, treatment with tannic acid increased the expression of Hsp70, Hsp90, and CYP, and heat shock proteins are more upregulated than the cyclophilin gene. Exposure to tannic acid increased the expression of stress factors over time (48 h) by 10- to 27-fold the expression level of the control group. These results suggest that tannic acid can be used to control harmful algal blooms such as those containing P. minimum in eutrophic coastal waters. PMID:27187440

  2. 2004 SWFWMD Citrus County Lidar Survey

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This metadata record describes the ortho & LIDAR mapping of Citrus County, FL. The mapping consists of LIDAR data collection, contour generation, and production...

  3. The aconitate hydratase family from Citrus

    Directory of Open Access Journals (Sweden)

    Cercos Manuel

    2010-10-01

    Full Text Available Abstract Background Research on citrus fruit ripening has received considerable attention because of the importance of citrus fruits for the human diet. Organic acids are among the main determinants of taste and organoleptic quality of fruits and hence the control of fruit acidity loss has a strong economical relevance. In citrus, organic acids accumulate in the juice sac cells of developing fruits and are catabolized thereafter during ripening. Aconitase, that transforms citrate to isocitrate, is the first step of citric acid catabolism and a major component of the citrate utilization machinery. In this work, the citrus aconitase gene family was first characterized and a phylogenetic analysis was then carried out in order to understand the evolutionary history of this family in plants. Gene expression analyses of the citrus aconitase family were subsequently performed in several acidic and acidless genotypes to elucidate their involvement in acid homeostasis. Results Analysis of 460,000 citrus ESTs, followed by sequencing of complete cDNA clones, identified in citrus 3 transcription units coding for putatively active aconitate hydratase proteins, named as CcAco1, CcAco2 and CcAco3. A phylogenetic study carried on the Aco family in 14 plant species, shows the presence of 5 Aco subfamilies, and that the ancestor of monocot and dicot species shared at least one Aco gene. Real-time RT-PCR expression analyses of the three aconitase citrus genes were performed in pulp tissues along fruit development in acidic and acidless citrus varieties such as mandarins, oranges and lemons. While CcAco3 expression was always low, CcAco1 and CcAco2 genes were generally induced during the rapid phase of fruit growth along with the maximum in acidity and the beginning of the acid reduction. Two exceptions to this general pattern were found: 1 Clemenules mandarin failed inducing CcAco2 although acid levels were rapidly reduced; and 2 the acidless "Sucreña" orange

  4. Establishment of Lipolexis oregmae (Hymenoptera: Aphidiidae) in a classical biological control program directed against the brown citrus aphid (Homoptera: Aphididae) in Florida

    International Nuclear Information System (INIS)

    The parasitoid Lipolexis oregmae Gahan (introduced as L. scutellaris Mackauer) was imported from Guam, evaluated in quarantine, mass reared, and released into citrus groves in Florida in a classical biological control program directed against the brown citrus aphid, Toxoptera citricida Kirkaldy. Releases of 20,200, 12,100, and 1,260 adults of L. oregmae were made throughout Florida during 2000, 2001, and 2002, respectively. To determine if L. oregmae had successfully established, surveys were conducted throughout the state beginning in the summer of 2001 and continuing through the summer of 2003. Parasitism during 2001 and 2002 was evaluated by holding brown citrus aphids in the laboratory until parasitoid adults emerged. Lipolexis oregmae was found in 10 sites in 7 counties and 4 sites in 3 counties with parasitism rates ranging from 0.7 to 3.3% in 2001 and 2002, respectively. Laboratory tests indicated that high rates of mortality occurred if field-collected parasitized aphids were held in plastic bags, so a molecular assay was used that allowed immature L. oregmae to be detected within aphid hosts immediately after collection. The molecular assay was used in 2003 with the brown citrus aphids and with other aphid species collected from citrus, weeds, and vegetables near former release sites; immatures of L. oregmae were detected in black citrus aphids, cowpea aphids, spirea aphids, and melon aphids, as well as in the brown citrus aphid, in 4 of 8 counties sampled, with parasitism ranging from 2.0 to 12.9%, indicating that L. oregmae is established and widely distributed. Samples taken in Polk County during Oct 2005 indicated that L. oregmae has persisted. The ability of L. oregmae to parasitize other aphid species on citrus, and aphids on other host plants, enhances the ability of L. oregmae to persist when brown citrus aphid populations are low. (author)

  5. Identification of a GCC transcription factor responding to fruit colour change events in citrus through the transcriptomic analyses of two mutants

    OpenAIRE

    Cercós Manuel; Zacarías Lorenzo; Alós Enriqueta; Rodrigo María-Jesús; Naranjo Miguel A; Ríos Gabino; Talón Manuel

    2010-01-01

    Abstract Background External ripening in Citrus fruits is morphologically characterized by a colour shift from green to orange due to the degradation of chlorophylls and the accumulation of carotenoid pigments. Although numerous genes coding for enzymes involved in such biochemical pathways have been identified, the molecular control of this process has been scarcely studied. In this work we used the Citrus clementina mutants 39B3 and 39E7, showing delayed colour break, to isolate genes poten...

  6. Protection from Metabolic Dysregulation, Obesity, and Atherosclerosis by Citrus Flavonoids: Activation of Hepatic PGC1α-Mediated Fatty Acid Oxidation

    OpenAIRE

    Mulvihill, Erin E.; Huff, Murray W.

    2012-01-01

    Studies in a multitude of models including cell culture, animal and clinical studies demonstrate that citrus-derived flavonoids have therapeutic potential to attenuate dyslipidemia, correct hyperinsulinemia and hyperglycemia, and reduce atherosclerosis. Emerging evidence suggests the metabolic regulators, PPARα and PGC1α, are targets of the citrus flavonoids, and their activation may be at least partially responsible for mediating their metabolic effects. Molecular studies will add significan...

  7. Evidence of Recombinant Citrus tristezavirus Isolate Occurring in Acid Lime cv. Pant Lemon Orchard in Uttarakhand Terai Region of Northern Himalaya in India

    OpenAIRE

    Singh, Jaywant Kumar; Tarafdar, Avijit; Sharma, Susheel Kumar; Biswas, Kajal Kumar

    2012-01-01

    The present study for the first time describes biological and molecular characterization of Citrus tristeza virus (CTV) occurring in the Terai area of Uttarakhand State in Northern Himalaya region of India. Direct antigen coated-ELISA and reverse transcriptase-polymerase chain reaction (RT-PCR) detected the CTV infection in Acid lime cv. Pant lemon (Citrus aurantifolia) orchards of Pantnagar with an estimated disease incidence of 16.6–20.5 %. To know the biological and genetic properties, an ...

  8. Comparative transcriptomics and proteomics analysis of citrus fruit, to improve understanding of the effect of low temperature on maintaining fruit quality during lengthy post-harvest storage

    OpenAIRE

    Yun, Ze; Jin, Shuai; Ding, Yuduan; Wang, Zhuang; Gao, Huijun; Pan, Zhiyong; Xu, Juan; Cheng, Yunjiang; Deng, Xiuxin

    2012-01-01

    Fruit quality is a very complex trait that is affected by both genetic and non-genetic factors. Generally, low temperature (LT) is used to delay fruit senescence and maintain fruit quality during post-harvest storage but the molecular mechanisms involved are poorly understood. Hirado Buntan Pummelo (HBP; Citrus grandis × C. paradis) fruit were chosen to explore the mechanisms that maintain citrus fruit quality during lengthy LT storage using transcriptome and proteome studies based on digital...

  9. Comparative transcriptome analysis of stylar canal cells identifies novel candidate genes implicated in the self-incompatibility response of Citrus clementina

    OpenAIRE

    Caruso Marco; Merelo Paz; Distefano Gaetano; La Malfa Stefano; Lo Piero Angela; Tadeo Francisco R; Talon Manuel; Gentile Alessandra

    2012-01-01

    Abstract Background Reproductive biology in citrus is still poorly understood. Although in recent years several efforts have been made to study pollen-pistil interaction and self-incompatibility, little information is available about the molecular mechanisms regulating these processes. Here we report the identification of candidate genes involved in pollen-pistil interaction and self-incompatibility in clementine (Citrus clementina Hort. ex Tan.). These genes have been identified comparing th...

  10. Antihyperlipidemic effects of Citrus sinensis, Citrus paradisi, and their combinations

    Directory of Open Access Journals (Sweden)

    Neelam Mallick

    2016-01-01

    Full Text Available Objective: Hyperlipidemia, extensively contributes in the progression of coronary heart diseases and atherosclerosis, but may be managed through alterations in the nutritional pattern. Several studies show that diet rich in polyphenols and antioxidants have antiatherogenic effects. Citrus sinensis and Citrus paradisi are widely known for health benefits and have found to produce antioxidant, anti-inflammatory, and hypolipidemic effects, hence current research was planned to determine the hypolipidemic effects of C. sinensis and C. paradisi in rats receiving diet rich in cholesterol. Materials and Methods: All rats were divided into 11 groups each comprising 10 animals: Normal control group and hyperlipidemic control. C. sinensis treated three groups, C. paradisi treated three groups, C. sinensis and C. paradisi combination treated two groups, and group treated atorvastatin. All rats in the respective groups were treated orally with sterile water, juices, and standard drug for 8 weeks and lipid profile was estimated at the end of dosing. Results: Cholesterol, triglycerides (TGs, and low-density lipoprotein (LDL were decreased at all the three doses of C. sinensis and C. paradisi but rise in high-density lipoprotein (HDL was only significant at 8 ml/kg, and 0.3 ml/kg, respectively. Animals received the combination doses of C. sinensis and C. paradisi also showed a highly significant fall in cholesterol, LDL, and TGs, however HDL level was significantly elevated by SPJ-2 combination. Conclusion: Results suggest that C. sinensis and C. paradisi possess antihyperlipidemic activity due to phytochemicals and other essential nutrients, hence may serve as cardioprotective by preventing thrombosis.

  11. Antihyperlipidemic effects of Citrus sinensis, Citrus paradisi, and their combinations

    Science.gov (United States)

    Mallick, Neelam; Khan, Rafeeq Alam

    2016-01-01

    Objective: Hyperlipidemia, extensively contributes in the progression of coronary heart diseases and atherosclerosis, but may be managed through alterations in the nutritional pattern. Several studies show that diet rich in polyphenols and antioxidants have antiatherogenic effects. Citrus sinensis and Citrus paradisi are widely known for health benefits and have found to produce antioxidant, anti-inflammatory, and hypolipidemic effects, hence current research was planned to determine the hypolipidemic effects of C. sinensis and C. paradisi in rats receiving diet rich in cholesterol. Materials and Methods: All rats were divided into 11 groups each comprising 10 animals: Normal control group and hyperlipidemic control. C. sinensis treated three groups, C. paradisi treated three groups, C. sinensis and C. paradisi combination treated two groups, and group treated atorvastatin. All rats in the respective groups were treated orally with sterile water, juices, and standard drug for 8 weeks and lipid profile was estimated at the end of dosing. Results: Cholesterol, triglycerides (TGs), and low-density lipoprotein (LDL) were decreased at all the three doses of C. sinensis and C. paradisi but rise in high-density lipoprotein (HDL) was only significant at 8 ml/kg, and 0.3 ml/kg, respectively. Animals received the combination doses of C. sinensis and C. paradisi also showed a highly significant fall in cholesterol, LDL, and TGs, however HDL level was significantly elevated by SPJ-2 combination. Conclusion: Results suggest that C. sinensis and C. paradisi possess antihyperlipidemic activity due to phytochemicals and other essential nutrients, hence may serve as cardioprotective by preventing thrombosis. PMID:27134462

  12. Characterization of the Asian Citrus Psyllid Transcriptome

    OpenAIRE

    Reese, Justin; Christenson, Matthew K.; Leng, Nan; Saha, Surya; Cantarel, Brandi; Lindeberg, Magdalen; Tamborindeguy, Cecilia; MacCarthy, Justin; Weaver, Daniel; Trease, Andrew J.; Ready, Steven V.; Davis, Vincent M.; McCormick, Courtney; Haudenschild, Christian; Han, Shunsheng

    2014-01-01

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is a vector for the causative agents of Huanglongbing, which threatens citrus production worldwide. This study reports and discusses the first D. citri transcriptomes, encompassing the three main life stages of D. citri, egg, nymph and adult. The transcriptomes were annotated using Gene Ontology (GO) and insecticide-related genes within each life stage were identified to aid the development of future D. citri insectici...

  13. Phyllosticta citriasiana sp nov., the cause of Citrus tan spot of Citrus maxima in Asia

    NARCIS (Netherlands)

    Wulandari, N.F.; To-anun, C.; Hyde, K.D.; Duong, L.M.; de Gruyter, J.; Meffert, J.P.; Groenewald, J.Z.; Crous, P.W.

    2009-01-01

    Guignardia citricarpa, the causal agent of Citrus Black Spot, is subject to phytosanitary legislation in the European Union and the U.S.A. This species is frequently confused with G. mangiferae, which is a non-pathogenic, and is commonly isolated as an endophyte from citrus fruits and a wide range o

  14. Phyllosticta citriasiana sp. nov., the cause of Citrus tan spot of Citrus maxima in Asia

    NARCIS (Netherlands)

    Wulandari, N.F.; To-anun, C.; Hyde, K.D.; Duong, L.M.; Gruyter, de J.; Meffert, J.P.; Groenewald, J.Z.; Crous, P.W.

    2009-01-01

    Guignardia citricarpa, the causal agent of Citrus Black Spot, is subject to phytosanitary legislation in the European Union and the U.S.A. This species is frequently confused with G. mangiferae, which is a non-pathogenic, and is commonly isolated as an endophyte from citrus fruits and a wide range o

  15. Detection of non-typhoid Salmonella infection by citrus and citrus extracts in Lao PDR

    Institute of Scientific and Technical Information of China (English)

    Yutaka Midorikawa; Satoshi Nakamura; Rattanaphone Phetsouvanh; Manivanh Vongsouvaht; Kaoru Midorikawa

    2010-01-01

    Objective: To know the current state of non-typhoid Salmonella infection in Laos. To examine the usefulness of new screening methods for Salmonella using citrus. Methods: Non-typhoidSalmonella infection of person in Lao PDR was studied in this research (2004-2009). The site was Vientiane capital city in 2004. Research from rural villages locating suburb of Vientiane during 2005-2008 was carried out. Rural villages in Attapu province where ethnic minorities were living was searched for this study in 2009. During this research, to detect Salmonella strain, a new method using citrus and citrus extract named MY phenomenon that observing black ring (MIDO ring) on DHL agar was tried. The slice lemon and lime were used for this trial in 2004. After 2005, disk of ascorbic acid and citric acid were used for the device instead of citrus fruits itself. Results: During this research, 65 of 272 human samples (23.9%) were infected with non-typhoid Salmonella. Conclusions: During this study, the method using citrus and citrus extracts was accepted for the detection of Salmonella. This study shows that with citrus and citrus extract, detection of Salmonella is possible using only DHL media. Results suggest that infectious rate of non-typhoid Salmonella was high.

  16. Evaluation of natural colorants and their application on citrus fruit as alternatives to citrus red II

    Science.gov (United States)

    The poor peel color of some varieties of oranges and the hybrids, especially for early season fruits, is caused by the subtropical climate of Florida, and has resulted in the use of a red dye on the peel to improve fruit appearance and marketability. Citrus Red II (CR2), the commercial citrus color ...

  17. Recovery and characterization of a Citrus clementina Hort. ex Tan. 'Clemenules' haploid plant selected to establish the reference whole Citrus genome sequence

    Directory of Open Access Journals (Sweden)

    Navarro Luis

    2009-08-01

    Full Text Available Abstract Background In recent years, the development of structural genomics has generated a growing interest in obtaining haploid plants. The use of homozygous lines presents a significant advantage for the accomplishment of sequencing projects. Commercial citrus species are characterized by high heterozygosity, making it difficult to assemble large genome sequences. Thus, the International Citrus Genomic Consortium (ICGC decided to establish a reference whole citrus genome sequence from a homozygous plant. Due to the existence of important molecular resources and previous success in obtaining haploid clementine plants, haploid clementine was selected as the target for the implementation of the reference whole genome citrus sequence. Results To obtain haploid clementine lines we used the technique of in situ gynogenesis induced by irradiated pollen. Flow cytometry, chromosome counts and SSR marker (Simple Sequence Repeats analysis facilitated the identification of six different haploid lines (2n = x = 9, one aneuploid line (2n = 2x+4 = 22 and one doubled haploid plant (2n = 2x = 18 of 'Clemenules' clementine. One of the haploids, obtained directly from an original haploid embryo, grew vigorously and produced flowers after four years. This is the first haploid plant of clementine that has bloomed and we have, for the first time, characterized the histology of haploid and diploid flowers of clementine. Additionally a double haploid plant was obtained spontaneously from this haploid line. Conclusion The first haploid plant of 'Clemenules' clementine produced directly by germination of a haploid embryo, which grew vigorously and produced flowers, has been obtained in this work. This haploid line has been selected and it is being used by the ICGC to establish the reference sequence of the nuclear genome of citrus.

  18. Dietary citrus pulp reduces lipid oxidation in lamb meat

    OpenAIRE

    Inserra, L.; Priolo, A.; L. Biondi; Lanza, M.; Bognanno, M.; Gravador, R.; Luciano, G.

    2014-01-01

    This study investigated the effect of replacing cereal concentrateswith high levels of dried citrus pulp in the diet on lamb meat oxidative stability. Over 56 days, lambs were fed a barley-based concentrate (Control) or concentrates inwhich 24% and 35% dried citrus pulpwere included to partially replace barley (Citrus 24% and Citrus 35%, respectively). Meat was aged under vacuum for 4 days and subsequently stored aerobically at 4 °C. The Control diet increased the redness, yellowness and satu...

  19. Whitefly Pest Species (Homoptera: Aleyrodidae) on Citrus Trees

    OpenAIRE

    Katja Žanić; Sonja Kačić; Miro Katalinić

    2000-01-01

    Today, the Citrus whitefly, Dialeurodes citri (Ashmead), is a very important pest on all Citrus species throughout the citrus growing areas in Croatia. It causes direct damage by sucking the plant juice from the leaves. Furthermore, immatures excrete honeydew that stimulates sooy mold. The presence of sooty mold on contaminated leaves interferes with the photosynthesis of plants. Citrus fruits coated by sooty mold lose its market value. Because Dialeurodes citri is poorly known in Croatia, th...

  20. Past and future of a century old Citrus tristeza virus collectionA California citrus germplasm tale

    Directory of Open Access Journals (Sweden)

    Jinbo eWang

    2013-12-01

    Full Text Available Citrus tristeza virus (CTV isolates collected from citrus germplasm, dooryard and field trees in California from 1914 have been maintained in planta under quarantine in the Citrus Clonal Protection Program (CCPP, Riverside, California. This collection, therefore, represents populations of CTV isolates obtained over time and space in California. To determine CTV genetic diversity in this context, genotypes of CTV isolates from the CCPP collection were characterized using multiple molecular markers (MMM. Genotypes T30, VT, and T36 were found at high frequencies with T30 and T30+VT genotypes being the most abundant. The MMM analysis did not identify T3 and B165/T68 genotypes; however, biological and phylogenetic analysis suggested some relationships of CCPP CTV isolates with these two genotypes. Phylogenetic analysis of the CTV coat protein gene sequences classified the tested isolates into seven distinct clades. Five clades were in association with the standard CTV genotypes T30, T36, T3, VT, and B165/T68. The remaining two identified clades were not related to any standard CTV genotypes. Spatiotemporal analysis indicated a trend of reduced genotype and phylogenetic diversity as well as virulence from southern California at early (1907-1957 in comparison to that of central California isolates collected from later (1957-2009 time periods. CTV biological characterization also indicated a reduced number and less virulent stem pitting (SP CTV isolates compared to seedling yellows isolates introduced to California. This data provides a historical insight of the introduction, movement, and genetic diversity of CTV in California and provides genetic and biological information useful for CTV quarantine, eradication, and disease management strategies such as CTV-SP cross protection.

  1. Current Situation of Citrus Huanglongbing in Guangdong, P. R. China

    Science.gov (United States)

    Guangdong Province is an important citrus production region in China. Citrus Huanglongbing (HLB, yellow shoot disease) was observed in Guangdong probably in the late 1800’s and the disease was first studied there. Since the 1990’s, citrus production in Guangdong has gradually shifted from the coasta...

  2. Intercropping of citrus and guava trees for management of Huanglongbing

    Science.gov (United States)

    Recent studies conducted in Viet Nam by Vietnamese, Japanese and Australian scientists indicate that interplanting citrus with guava negated infestations of Asian citrus psyllid and consequently huanglongbing, a serious disease caused by a bacterium vectored by the psyllid. Young citrus interplanted...

  3. Trailers transporting oranges to processing plants move Asian citrus psyllids

    Science.gov (United States)

    Huanglongbing (citrus greening) is one of the most serious of citrus diseases. Movement of the disease occurs as a result of natural vector-borne infection and by movement of plant material. We demonstrate here that Diaphorina citri Kuwayama (vector of citrus greening pathogens) can be transported i...

  4. Characterizing the citrus variety Carrizo genome through 454 shotgun sequencing

    Science.gov (United States)

    Citrus production is of global importance both in economic impact and significance to nutrition. The number of natural citrus species appears extremely limited. The genome size is small (haploid approximately 367 Mb), arranged on 18 chromosomes. The citrus variety Carrizo, generated by a ‘Washingt...

  5. First report of citrus leprosis virus nuclear type in Colombia

    Science.gov (United States)

    Citrus leprosis is a difficult viral disease causing significant damage to citrus fruit in South America and Central America. The disease is marked by dramatic lesions on fruit, leaves and stems resulting in unmarketable product. Citrus leprosis virus cytoplasmic types (CiLV-C and CiLV-C2) wer edete...

  6. Evaluation of natural colorants and their application on citrus fruit

    Science.gov (United States)

    Warm temperatures can often result in poor peel color of some citrus varieties, especially early in the harvest season. Under these conditions, Florida oranges, temples, tangelos, and K-Early citrus fruit are allowed to be treated with Citrus Red No.2 (CR2) to help produce a more acceptable peel col...

  7. Thirty years of citrus tristeza virus observations in Peru

    Science.gov (United States)

    The Peruvian citrus industry was devastated by epidemics of Citrus tristeza virus (CTV) decline (CTV-D) on sour orange rootstock between 1950 and 1965 and CTV stem pitting (SP) between 1965 and 1985. CTV-SP debilitates citrus and fruit production regardless of rootstock. Control of CTV-SP by mild st...

  8. Expression and variability of molecular chaperones in the sugarcane expressome.

    Science.gov (United States)

    Borges, Júlio C; Cagliari, Thiago C; Ramos, Carlos H I

    2007-04-01

    Molecular chaperones perform folding assistance in newly synthesized polypeptides preventing aggregation processes, recovering proteins from aggregates, among other important cellular functions. Thus their study presents great biotechnological importance. The present work discusses the mining for chaperone-related sequences within the sugarcane EST genome project database, which resulted in approximately 300 different sequences. Since molecular chaperones are highly conserved in most organisms studied so far, the number of sequences related to these proteins in sugarcane was very similar to the number found in the Arabidopsis thaliana genome. The Hsp70 family was the main molecular chaperone system present in the sugarcane expressome. However, many other relevant molecular chaperones systems were also present. A digital RNA blot analysis showed that 5'ESTs from all molecular chaperones were found in every sugarcane library, despite their heterogeneous expression profiles. The results presented here suggest the importance of molecular chaperones to polypeptide metabolism in sugarcane cells, based on their abundance and variability. Finally, these data have being used to guide more in deep analysis, permitting the choice of specific targets to study. PMID:16687190

  9. Citrus plant nutritional profile in relation to huanglongbing prevalence in Pakistan

    International Nuclear Information System (INIS)

    Citrus is an important fruit crop in Pakistan that requires proper crop nutrition and disease management strategies as it is a tree crop and withstands harsh seasonal conditions for decades. Huanglongbing (HLB) is a century old, devastating disease of citrus caused by phloem limiting bacteria of the alpha-proteobacteria subdivision. As disease has no known cure, so, effective prevention methods are useful in crop management. Improper crop nutrition impairs plant genetic resistance to invasive pathogens, decreases yield and reduces productive life of the plant. In this study we selected 116 citrus trees from 43 orchard of Punjab for a nutritional assessment. All the trees were showing HLB symptoms and were subjected to NPK and Zn analysis as well as molecular detection of Candidatus L. asiaticus, the pathogen associated with HLB. Nitrogen and Zn were significantly higher (P=0.05) in HLB infected trees. Out of 48 diseased trees, 19, 43 and 27 were deficient in nitrogen, phosphorous and potash, respectively. Our study concludes that there is no relationship between nutritional deficiency status and HLB incidence in citrus; however, nutritional treatments may help in stress relief to infected plants. (author)

  10. Complete genome sequence analysis of two Citrus tatter leaf virus (CTLV) isolates from China

    Institute of Scientific and Technical Information of China (English)

    SONG Zhen; LI Zhong-an; LIU Ke-hong; ZHOU Chang-yong

    2015-01-01

    In order to understand molecular characterization of Citrus tatter leaf virus (CTLV) isolated from China, ful-length cDNAs of CTLV-MTH and CTLV-XHC from Citrus reticulata and Citrus sinensis were cloned and sequenced based on whole-genome ampliifcation by RT-PCR. The complete nucleotide sequences of CTLV-MTH and CTLV-XHC were determined to be 6 497 nucleotides in length and shared 79.9–91.0%and 78.8–98.0%nucleotide sequence identity, respectively, with other Apple stem grooving virus (ASGV) or CTLV strains available in GenBank. Unexpectedly, CTLV-MTH showed the highest nucleo-tide sequence identity (91%) with an apple isolate of ASGV, fol owed by 86.5%with ASGV-HH and 85.7%with ASGV-CHN. Furthermore, CTLV-MTH and three ASGV strains were grouped to a separate cluster in the phylogenetic tree, suggesting it has a closer relationship to ASGV than to CTLV. Therefore, it can be concluded roughly that CTLV may be not a distinct strains of ASGV. We proposed that Citrus tatter leaf virus should be renamed Apple stem grooving virus.

  11. Isolation and Characterization of Copia-like Retrotransposons from 12 Sweet Orange (Citrus sinensis Osbeck) Cultivars

    Institute of Scientific and Technical Information of China (English)

    Neng-Guo TAO; Juan XU; Yun-Jiang CHENG; Liu HONG; Wen-Wu GUO; Hua-Lin YI; Xiu-Xin DENG

    2005-01-01

    As the largest transposable element in the plant genome, retrotransposons are thought to be involved in citrus genetic instability and genome evolution, especially in sweet orange, which is prone to bud mutation. In the present study, the presence of copia-like retrotransposons, their heterogeneity, genomic distribution, and transcriptional activities in Citrus were investigated in 12 sweet orange (Citrus sinensis Osbeck) cultivars using a PCR assay designed to detect copia-like reverse transcriptase (RT)sequences. Twelve amplification products from each cultivar were cloned and sequenced. The cloned sequences showed great heterogeneity, except "Dream" navel and "Hamlin", both of which shared the same sequence. Frame shifting, termination, deletion, and substitution accounted for the heterogeneity of RT sequences. Southern blot hybridization using the RT1 clone from the "Cara Cara" navel as a probe showed that multiple copies were integrated throughout the sweet orange genomes, which made the retrotransposon possible an effective molecular marker to detect citrus evolution events and to reveal its relationship with bud mutation. No transcriptional activities of the retrotransposon were detected by RT-PCR and Northern analysis in the fruits and leaves of either "Cara Cara" or "Seike" navels.

  12. Does citrus leaf miner impair hydraulics and fitness of citrus host plants?

    Science.gov (United States)

    Raimondo, Fabio; Trifilò, Patrizia; Gullo, Maria A Lo

    2013-12-01

    Gas exchange and hydraulic features were measured in leaves of three different Citrus species (Citrus aurantium L., Citrus limon L., Citrus  ×  paradisii Macfad) infested by Phyllocnistis citrella Staiton, with the aim to quantify the impact of this pest on leaf hydraulics and, ultimately, on plant fitness. Infested leaves were characterized by the presence on the leaf blade of typical snake-shaped mines and, in some cases, of a crumpled leaf blade. Light microscopy showed that leaf crumpling was induced by damage to the cuticular layer. In all three Citrus species examined: (a) the degree of infestation did not exceed 10% of the total surface area of infested plants; (b) control and infested leaves showed similar values of minimum diurnal leaf water potential, leaf hydraulic conductance and functional vein density; and (c) maximum diurnal values of stomatal conductance to water vapour, transpiration rate and photosynthetic rate (An) were similar in both control leaves and the green areas of infested leaves. A strong reduction of An was recorded only in mined leaf areas. Our data suggest that infestation with P. citrella does not cause conspicuous plant productivity reductions in young Citrus plants, at least not in the three Citrus species studied here. PMID:24319027

  13. Evaluation of natural colorants and their application on citrus fruit as alternatives to Citrus Red No. 2

    Science.gov (United States)

    Warm field temperatures can often result in poor peel color of some citrus varieties, especially early in the harvest season. Under these conditions, Florida oranges, temples, tangelos, and K-Early citrus fruit are allowed to be treated with Citrus Red No.2 (CR2) to help produce a more acceptable pe...

  14. Isolation of Xylella fastidiosa from Citrus sinensis (L) Osb. And Vitis vinifera and study of genetic diversity in Costa Rica

    International Nuclear Information System (INIS)

    Xylella fastidiosa is a plant pathogenic bacterium that causes diseases in different crops. Symptoms similar to those caused by citrus variegated chlorosis (CVC) were observed in sweet orange trees which served as shade and fences in coffee plantations in Costa Rica, in 2002. A total of 35 citrus trees and 24 vines from eight different districts and 3 respectively were evaluated by 'double antibody sandwich enzyme-linked immunosorbent assay' (DASELISA), resulting in 21 citrus and 19 positive vid. From four citrus trees and six of vines, were obtained six isolates and seven isolates respectively in solid medium, whose morphological and biochemical characteristics coincided with those reported in the literature as characteristic of X. fastidiosa. The identity of the isolates is confirmed by the chain polymerase reaction (PCR) using primers 272-1/272-2int and RST31/RST33. Three isolates from Grecia (Alajuela Province) amplified a band of 500pb using specific primers 272-2int/CVC-1 for strains of X. fastidiosa that cause CVC. The genetic variability of isolates from each other in comparison with isolates of coffee in Costa Rica, U.S. grapes and citrus in Brazil have been studied using techniques of random amplification polymorphism DNA (RAPD) and length polymorphisms of restriction fragments (RFLPs) of the products obtained with primers int/272-2int JB-1/JB-2 and 272-1. The results showed a clear separation between citrus isolates of Costa Rica; and, an association of three of them with the strains of citrus in Brasil. Also, an association between strains of coffee of Costa Rica with grape vines in the U.S. An association of molecular analysis confirmed the data variance. (author)

  15. Effect of Citrus Flavonoids, Naringin and Naringenin, on Metabolic Syndrome and Their Mechanisms of Action12

    OpenAIRE

    Alam, M. Ashraful; Subhan, Nusrat; Rahman, M. Mahbubur; Uddin, Shaikh J.; Reza, Hasan M.; Satyajit D. Sarker

    2014-01-01

    Flavonoids are important natural compounds with diverse biologic activities. Citrus flavonoids constitute an important series of flavonoids. Naringin and its aglycone naringenin belong to this series of flavonoids and were found to display strong anti-inflammatory and antioxidant activities. Several lines of investigation suggest that naringin supplementation is beneficial for the treatment of obesity, diabetes, hypertension, and metabolic syndrome. A number of molecular mechanisms underlying...

  16. Sequence diversity on four ORFs of citrus tristeza virus correlates with pathogenicity

    OpenAIRE

    Martínez-Soriano Juan; Rivera-Bustamante Rafael; Ochoa-Sánchez Juan; Herrera-Isidrón Lisset

    2009-01-01

    Abstract The molecular characterization of isolates of citrus tristeza virus (CTV) from eight locations in Mexico was undertaken by analyzing five regions located at the opposite ends of the virus genome. Two regions have been previously used to study CTV variability (coat protein and p23), while the other three correspond to other genomic segments (p349-B, p349-C and p13). Our comparative nucleotide analyses included CTV sequences from different geographical origins already deposited in the ...

  17. Detection of citrus canker and Huanglongbing using fluorescence imaging spectroscopy and support vector machine technique.

    Science.gov (United States)

    Wetterich, Caio Bruno; Felipe de Oliveira Neves, Ruan; Belasque, José; Marcassa, Luis Gustavo

    2016-01-10

    Citrus canker and Huanglongbing (HLB) are citrus diseases that represent a serious threat to the citrus production worldwide and may cause large economic losses. In this work, we combined fluorescence imaging spectroscopy (FIS) and a machine learning technique to discriminate between these diseases and other ordinary citrus conditions that may be present at citrus orchards, such as citrus scab and zinc deficiency. Our classification results are highly accurate when discriminating citrus canker from citrus scab (97.8%), and HLB from zinc deficiency (95%). These results show that it is possible to accurately identify citrus diseases that present similar symptoms. PMID:26835778

  18. Association of Plasma IL-6 and Hsp70 with HRV at Different Levels of PAHs Metabolites

    OpenAIRE

    Jian Ye; Rui Zhu; Xiaosheng He; Yingying Feng; Liangle Yang; Xiaoyan Zhu; Qifei Deng; Tangchun Wu; Xiaomin Zhang

    2014-01-01

    BACKGROUND: Exposure to polycyclic aromatic hydrocarbons (PAHs) is associated with reduced heart rate variability (HRV), a strong predictor of cardiovascular diseases, but the mechanism is not well understood. OBJECTIVES: We hypothesized that PAHs might induce systemic inflammation and stress response, contributing to altered cardiac autonomic function. METHODS: HRV indices were measured using a 3-channel digital Holter monitor in 800 coke oven workers. Plasma levels of interleukin-6 (IL-6) a...

  19. Hsp70 protects mitotic cells against heat-induced centrosome damage and division abnormalities

    NARCIS (Netherlands)

    Hut, HMJ; Kampinga, HH; Sibon, OCM

    2005-01-01

    The effect of heat shock on centrosomes has been mainly studied in interphase cells. Centrosomes play a key role in proper segregation of DNA during mitosis. However, the direct effect and consequences of heat shock on mitotic cells and a possible cellular defense system against proteotoxic stress d

  20. Immune response to p53 is dependent upon p53/HSP70 complexes in breast cancers.

    OpenAIRE

    Davidoff, A.M.; Iglehart, J D; Marks, J R

    1992-01-01

    Overexpression of the p53 protein, resulting from gene mutations that increase protein stability, has been detected in greater than 25% of primary human breast cancers. In addition, approximately 10% of breast cancer patients have circulating antibodies to the p53 protein. In this study, the anti-p53 humoral response is correlated with the presence and type of mutant p53 protein expressed in the tumor. In a series of 60 breast cancer patients, 0 of 30 tumors with normal, low-level p53 express...

  1. HSP70与恶性肿瘤关系的研究进展

    Institute of Scientific and Technical Information of China (English)

    游朝勇

    2012-01-01

    @@ 热休克蛋白(heat shock proteins,HSPs)又称应激蛋白(stress protein,sp),是生物体细胞在热诱导下合成的一组具有高度保守性的蛋白质[1].近年来,HSPs被发现与恶性肿瘤的发生、发展、复发及预后关系密切.

  2. 70 kD stress protein (Hsp70) analysis in living shallow-water benthic foraminifera

    Digital Repository Service at National Institute of Oceanography (India)

    Heinz, P.; Marten, R.A.; Linshy, V.N.; Haap, T.; Geslin, E.; Kohler, H-R.

    of Evolution and Ecology, University of Tübingen, Konrad- Adenauer-Str. 20, 72072 Tübingen, Germany 4 Laboratoire d`Etude des Bio-indicateurs Actuels et Fossiles (BIAF) and LEBIM, University of Angers, 2 Boulevard Lavoisier, 49045 Angers Cedex, France... beccarii and Elphidium crispum were collected on the rocky shore near the Laboratory of Marine Bio-Indicators (LEBIM) on Yeu Island (France) in July 2007 and May 2008. Ammonia tepida was collected in the muddy shore in the Bay of Aiguillon (France...

  3. Dynamics of endogenous Hsp70 synthesis in the brain of olfactory bulbectomized mice

    OpenAIRE

    Bobkova, Natalia; Guzhova, Irina; Margulis, Boris; Nesterova, Inna; Medvedinskaya, Natalia; Samokhin, Alexander; Alexandrova, Irina; Garbuz, David; Nudler, Evgeny; Evgen’ev, Michael

    2012-01-01

    Numerous epidemiological studies have established acute brain injury as one of the major risk factors for the Alzheimer's disease (AD). However, the lack of animal models of AD-like degeneration triggered by a defined injury hampered the development of adequate therapies. Here we report that the surgical damage of the olfactory bulbs triggers the development of several pathologies, including amyloid-β accumulation and strong decrease of neuron density in the cortex and hippocampus as well as ...

  4. Noninvasive maturity detection of citrus with machine vision

    Science.gov (United States)

    Ying, Yibin; Xu, Zhenggang; Fu, Xiaping; Liu, Yande

    2004-03-01

    A computer vision system was established to explore a method for citrus maturity detection. The surface color information and the ratio of total soluble solid to titratable acid (TSS/TA) were used as maturity indexes of citrus. The spectral reflectance properties with different color were measured by UV-240 ultraviolet and visible spectrophotometer. The biggest discrepancy of gray levels between citrus pixels and background pixels was in blue component image by image background segmentation. Dynamic threshold method for background segmentation had best result in blue component image. Methods for citrus image color description were studied. The citrus spectral reflectance experiments showed that green surface and saffron surface of citrus were of highest spectral reflectance at the wavelength of 700nm, the difference between them reached to maximum, about 53%, and the image acquired at this wavelength was of more color information for maturity detection. A triple-layer feed forward network was established to map citrus maturity from the hue frequency sequence by the mean of artificial neural network. After training, the network mapper was used to detect the maturity of the test sample set, which was composed of 252 Weizhang citrus with different maturity. The identification accuracy of mature citrus reached 79.1%, that of immature citrus was 63.6%, and the mean identification accuracy was 77.8%. This study suggested that it is feasible to detect citrus maturity non-invasively by using the computer vision system and hue frequency sequence method.

  5. Next generation haplotyping to decipher nuclear genomic interspecific admixture in Citrus species: analysis of chromosome 2

    OpenAIRE

    Curk, Franck; Ancillo, Gema; Garcia-Lor, Andres; Luro, François; Perrier, Xavier; Jacquemoud-Collet, Jean-Pierre; Navarro, Luis; Ollitrault, Patrick

    2014-01-01

    Background The most economically important Citrus species originated by natural interspecific hybridization between four ancestral taxa (Citrus reticulata, Citrus maxima, Citrus medica, and Citrus micrantha) and from limited subsequent interspecific recombination as a result of apomixis and vegetative propagation. Such reticulate evolution coupled with vegetative propagation results in mosaic genomes with large chromosome fragments from the basic taxa in frequent interspecific heterozygosity....

  6. 7 CFR 457.107 - Florida citrus fruit crop insurance provisions.

    Science.gov (United States)

    2010-01-01

    ... of the following: (1) Citrus I—Early and mid-season oranges; (2) Citrus II—Late oranges juice; (3) Citrus III—Grapefruit for which freeze damage will be adjusted on a juice basis; (4) Citrus IV—Tangelos...; or (2) The following juice content, if acceptable records are not furnished: (i) Citrus I—52...

  7. Biogenic emissions from Citrus species in California

    Science.gov (United States)

    Fares, Silvano; Gentner, Drew R.; Park, Jeong-Hoo; Ormeno, Elena; Karlik, John; Goldstein, Allen H.

    2011-09-01

    Biogenic Volatile Organic Compounds (BVOC) emitted from plants are the dominant source of reduced carbon chemicals to the atmosphere and are important precursors to the photochemical production of ozone and secondary organic aerosols. Considering the extensive land used for agriculture, cultivated Citrus plantations may play an important role in the chemistry of the atmosphere especially in regions such as the Central Valley of California. Moreover, the BVOC emissions from Citrus species have not been characterized in detail and more species-specific inputs for regional models of BVOC emissions are needed. In this study, we measured the physiological parameters and emissions of the most relevant BVOC (oxygenated compounds, monoterpenes, and sesquiterpenes) for four predominant Citrus species planted in California ( Citrus sinensis var. 'Parent Navel', Citrus limon var. 'Meyer', Citrus reticulata var. 'W. Murcott' and 'Clementine'). We used two analytical techniques to measure a full range of BVOC emitted: Proton Transfer Reaction Mass Spectrometry (PTR-MS) and gas chromatography with mass spectrometry. Methanol, followed by acetone and acetaldehyde, were the dominant BVOC emitted from lemon and mandarin trees (basal emission rates up to 300 ng(C) g(DW) -1 h -1), while oxygenated monoterpenes, monoterpenes, and sesquiterpenes were the main BVOC emitted from orange trees (basal emission rates up to = 2500 ng(C) g(DW) -1 h -1). Light and temperature-dependent algorithms were better predictors of methanol, acetaldehyde, acetone, isoprene and monoterpenes for all the Citrus species. Whereas, temperature-dependent algorithms were better predictors of oxygenated monoterpenes, and sesquiterpenes. We observed that flowering increased emissions from orange trees by an order of magnitude with the bulk of BVOC emissions being comprised of monoterpenes, sesquiterpenes, and oxygenated monoterpenes. Chemical speciation of BVOC emissions show that the various classes of terpene

  8. Rootstock-scion interaction affecting citrus response to CTV infection: a proteomic view.

    Science.gov (United States)

    Laino, Paolo; Russo, Maria P; Guardo, Maria; Reforgiato-Recupero, Giuseppe; Valè, Giampiero; Cattivelli, Luigi; Moliterni, Vita M C

    2016-04-01

    Citrus tristeza virus (CTV) is the causal agent of various diseases with dramatic effects on citrus crops worldwide. Most Citrus species, grown on their own roots, are symptomless hosts for many CTV isolates. However, depending on different scion-rootstock combination, CTV infection should result in distinct syndromes, being 'tristeza' the more severe one, leading to a complete decline of the susceptible plants in a few weeks. Transcriptomic analyses revealed several genes involved either in defense response, or systemic acquired resistance, as well as transcription factors and components of the phosphorylation cascades, to be differentially regulated during CTV infection in Citrus aurantifolia species. To date little is known about the molecular mechanism of this host-pathogen interaction, and about the rootstock effect on citrus response to CTV infection. In this work, the response to CTV infection has been investigated in tolerant and susceptible scion-rootstock combinations by two-dimensional gel electrophoresis (2DE). A total of 125 protein spots have been found to be differently accumulated and/or phosphorylated between the t